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Sample records for labdanic diterpenes estudo

  1. Dimeric Labdane Diterpenes: Synthesis and Antiproliferative Effects

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    Guillermo Schmeda-Hirschmann

    2013-05-01

    Full Text Available Several diterpenes with the labdane skeleton show biological activity, including antiproliferative effects. Most of the research work on bioactive labdanes has been carried out on naturally occurring diterpenes and semisynthetic derivatives, but much less is known on the effects of diterpene dimers. The aim of the present work was to synthesize dimeric diterpenes from the labdane imbricatolic acid using esters, ethers and the triazole ring as linkers. Some 18 new derivatives were prepared and the compounds were evaluated for antiproliferative activity on human normal fibroblasts (MRC-5 and the following human tumor cell lines: AGS, SK-MES-1, J82 and HL-60. The diethers 8–10, differing in the number of CH2 units in the linker, presented better antiproliferative activity with a maximum effect for the derivative 9. The best antiproliferative effect against HL-60 cells was found for compounds 3 and 17, with IC50 values of 22.3 and 23.2 μM, lower than that found for the reference compound etoposide (2.23 μM. The compounds 9, 17 and 11 were the most active derivatives towards AGS cells with IC50 values of 17.8, 23.4 and 26.1 μM. A free carboxylic acid function seems relevant for the effect as several of the compounds showed less antiproliferative effect after methylation.

  2. New labdane diterpenes from Leonurus cardiaca.

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    Agnihotri, Vijai K; Elsohly, Hala N; Smillie, Troy J; Khan, Ikhlas A; Walker, Larry A

    2008-08-01

    Leonurus cardiaca L. has been used in oriental medicine against several types of disorders. The ethanolic extract of leaves of Leonurus cardiaca yielded three new labdane-type diterpenes: 15- O-ethylleopersin C (1), 15- O-methylleopersin C (2), and 15- EPI- O-methylleopersin C (3). Their structures were determined using 1 D and 2 D NMR including 1H-1H COSY, HMQC, HMBC, and ROESY spectroscopic techniques. Compounds (1 - 3) were evaluated for in vitro antiplasmodial activity (D6 and W2 clones) and cytotoxicity (Vero cells).

  3. Accumulation of furanic labdane diterpenes in Marrubium vulgare and Leonurus cardiaca.

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    Knöss, W; Zapp, J

    1998-05-01

    Accumulation of furanic labdane diterpenes has been investigated in different parts of field-grown plants of MARRUBIUM VULGARE (Lamiaceae) and LEONURUS CARDIACA (Lamiaceae). Furanic labdane diterpenes were produced and accumulated only in the aerial parts. Greatest amounts were measured in leaves and flowers. Up to 4 mg furanic labdane diterpenes per g fresh weight were found. Accumulation of furanic labdane diterpenes in plantlets seemingly depends on a developmental programme. No furanic labdane diterpenes were detected in plantlets during the first four to five weeks following germination. At this time the leaves became more differentiated and the number of trichomes on leaves was obviously increasing. Young leaves and buds contained most furanic labdane diterpenes. It was proven that at least a part of the non-volatile furanic labdane diterpenes is stored in peltate glandular trichomes. NMR signals of marrubiin were investigated with correlated spectra. Some (1)H- and (13)C-NMR assignments reported in literature were revised.

  4. Chlorophytoside A, a New Labdane Diterpene Glycoside from Chlorophytum laxum

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    You Heng GAO; Quan Xi MEI; Hui Fei WU; Yuan Ming DIAO; Xin Sheng PENG; Rui Fang ZHANG

    2005-01-01

    A new labdane-type diterpene glycoside 1 , chlorophytoside A, had been isolated from Chlorophytum laxum R.Br. The structure had been elucidated as (10S)-6α-hydroxylabda-8,13-dien-15,16-olide 3R-O-β-D-glucopyranoside on the basis of chemical and spectroscopic data.

  5. Two Labdane Diterpenes from Leonurus heterophyllus

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    Xian ZHANG; Guo Ping PENG

    2006-01-01

    Two new labdane diterpenoids named heteronone A (1) and heteronone B (2) were isolated from the aerial parts of Leonurus heterophyllus Sweet. Their structures were established by various spectroscopic means, and the structure of heteronone A is confirmed by X-ray diffraction analysis.

  6. A new labdane diterpene from the rhizomes of Alpinia officinarum.

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    Zou, Qiong-Yu; Wu, Hai-Feng; Tang, Yu-Lian; Chen, Di-Zhao

    2016-01-01

    A new labdane diterpene, (Z)-12,14-labdadien-15(16)-olide-17-oic acid (1), and a new natural cadinane sesquiterpene, 4-isopropyl-6-methyl-1-naphthalenemethanol (2), were isolated from the ethanolic extract of the rhizomes of Alpinia officinarum, together with three other products, galangin (3), kaempferol (4) and quercetin (5). Their structures were elucidated by using extensive spectroscopic methods. Compounds 1 and 2 showed no remarkable cytotoxic activity against HeLa and HepG2 cancer cell lines with IC50>50 μg mL(- 1).

  7. Two New Labdane Diterpene Glycoside from Flowers of Bacchris Medulosa DC

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    D. A. Cifuente

    2000-03-01

    Full Text Available Two new labdane-type diterpene glycoside, were isolated from the flowers of Baccharis medulosa DC (Asteraceae. Structures of these compounds were established by application of various spectroscopic techniques.

  8. New Labdane Diterpenes and Their Glycoside Derivatives from the Roots of Isodon adenantha.

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    Wu, La-Bin; Xiao, Chao-Jiang; Jiang, Xue; Qiu, Lin; Dong, Xiang; Jiang, Bei

    2015-08-01

    Two new labdane-type diterpenes (adenanthic acids A and B; 1 and 2, resp.) and three new labdane diterpene glycosides (adenanthosides A-C; 3-5, resp.) were isolated from the roots of Isodon adenantha, together with 23 known constituents including seven diterpenoids (6-12), eight triterpenoids (13-20), one lignan glycoside (21), six steroids (22-27), and one ceramide (28). Their structures were elucidated by spectroscopic methods including extensive 2D-NMR techniques. Cytotoxicity and antibacterial activities of the samples were measured by the MTT method and the filter paper disc agar diffusion method. But none of them showed significant activities.

  9. Labdane-type diterpenes active against acne from pine cones (Pinus densiflora).

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    Sultan, Md Zakir; Jeon, Young-Min; Moon, Surk-Sik

    2008-03-01

    Bioassay-guided extraction and fractionation of the aqueous methanolic extract of the cones of Pinus densiflora (Pinaceae) afforded one new labdane-type diterpene aldehyde, 15-nor-14-oxolabda-8(17),12 E-diene-18-oic acid, along with eight known diterpenes. Their structures were elucidated using spectroscopic methods as well as by comparison with previously reported data. The isolates showed antibacterial (Propionibacterium acnes) and antifungal activities.

  10. Diterpene lactones with labdane, halimane and clerodane frameworks.

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    Silva, Lúcia; Gomes, Arlindo C; Rodilla, Jesus M L

    2011-04-01

    The labdane, halimane and clerodane type diterpenoids are compounds that have been isolated in plants of several families. These molecules and their derivatives with a lactone group on the side chain or on the decaline system, have a great interest because of their biological properties as insect antifeedant, antiviral, cytotoxic and trypanocidal. The scope of this review is lactones diterpenoids with labdane, halimane and clerodane frameworks.

  11. A new labdane diterpene from Vitex cauliflora Moldenke from the Madagascar rainforest1

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    Rasamison, Vincent E.; Ranaivo-Harimanana, Leon; Cao, Shugeng; Pan, Ende; Ratovoson, Fidy; Randriantafika, F.; Rakotondrajaona, R.; Rakotonandrasana, Stephan; Andriantsiferana, Rabodo

    2009-01-01

    Fractionation of an antiplasmodial ethanolic extract from the aerial parts of Vitex cauliflora led to the isolation of the new labdane diterpene 1 together with the known triterpene uvaol. The structure of the new compound 1 was established as 3-oxo,15,17,18-triacetoxy-labda-7,13E-diene on the basis of spectroscopic data (1D and 2D NMR, MS). PMID:19635529

  12. Biosynthesis of the labdane diterpene marrubiin in Marrubium vulgare via a non-mevalonate pathway.

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    Knöss, W; Reuter, B; Zapp, J

    1997-09-01

    The biosynthesis of the furanic labdane diterpene marrubiin has been studied in plantlets and shoot cultures of Marrubium vulgare (Lamiaceae). The use of [2-14C]acetate, [2-14C]pyruvate, [2-14C]mevalonic acid and [U-14C]glucose incorporation experiments showed that the labelling of sterols in etiolated shoot cultures of M. vulgare was in accordance with their biosynthesis via the acetate-mevalonate pathway. In contrast, the incorporation rates of these precursors into the diterpene marrubiin could not be explained by biosynthesis of this compound via the acetate-mevalonate pathway. Cultivation of etiolated shoot cultures of M. vulgare on medium containing [1-13C]glucose and subsequent 13C-NMR spectroscopy of marrubiin led to the conclusion that the biosynthesis of marrubiin follows a non-mevalonate pathway. All isoprenic units of 13C-labelled marrubiin were enriched in those carbons that correspond to positions 1 and 5 of a putative precursor isopentenyl diphosphate. This labelling pattern from [1-13C]glucose is consistent with an alternative pathway via trioses, which has already been shown to occur in Eubacteria and Gymnospermae. The labdane skeleton is a precursor of many other skeletal types of diterpenes. Therefore it becomes obvious that in connection with the few known examples of a non-mevalonate pathway to isoprenoids the formation of some isoprenoids in plants via a non-mevalonate pathway might be quite common.

  13. Synthesis of new ent-labdane diterpene derivatives from andrographolide and evaluation on cytotoxic activities.

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    Luo, Yan; Wang, Ke; Zhang, Meng-han; Zhang, Da-yong; Wu, Yang-chang; Wu, Xiao-ming; Hua, Wei-yi

    2015-06-01

    There are many reports for andrographolide modification regarding antitumor effects. Transformation of the five-membered lactone ring to furan aromatic ring still results in compounds with good cytotoxicity. To determine further the importance of the five-membered lactone ring and to obtain better lead compounds, we transformed the five-membered lactone ring in andrographolide. New types of ent-labdane diterpene derivatives were made, whose cytotoxic activities were measured in vitro. Preliminary SAR was summarized and two compounds, 7 and 26, with good cytotoxic activity were obtained, which have the potential to be developed into new antitumor drugs.

  14. Chemical diversity of labdane-type bicyclic diterpene biosynthesis in Actinomycetales microorganisms.

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    Yamada, Yuuki; Komatsu, Mamoru; Ikeda, Haruo

    2016-07-01

    Five pairs of bacterial type-A and type-B diterpene synthases have been characterized: BAD86798/BAD86797, AHK61133/AHK61132, BAB39207/BAB39206, CldD/CldB and RmnD/RmnB, and are involved in the formation of pimara-9(11),15-diene, terpente-3,13,15-triene and labda-8(17),12(E),14-triene. Mining of bacterial genome data revealed an additional four pairs of type-A and type-B diterpene synthases: Sros_3191/Sros_3192 of Streptosporangium roseum DSM 43021, Sare_1287/Sare_1288 of Salinispora arenicola CNS-205, SCLAV_5671/SCLAV_5672 and SCLAV_p0491/SCLAV_p0490 of Streptomyces clavuligerus ATCC 27064. Since SCLAV_p0491/SCLAV_p0490 is similar to the labdane-type diterpene synthase pairs, CldD/CldB and RmnD/RmnB based on the alignment of the deduced amino acid sequences and phylogenetic analyses of the aligned sequences, these predicted diterpene synthases were characterized by an enzymatic reaction using a pair of recombinant type-A and type-B diterpene synthases prepared in Escherichia coli and the heterologous expression of two genes encoding type-A and type-B diterpene synthases in an engineered Streptomyces host. The generation of labda-8(17),12(E),14-triene (1) by CldB and CldD was reconfirmed by enzymatic synthesis. Furthermore, labda-8(17),13(16),14-triene (2) was generated by SCLAV_p0491 and CldB, and ladba-7,12(E),14-triene (3) by CldD and SCLAV_p0490. SCLAV_p0491 and SCLAV_p0490 catalyzed the generation of the novel diterpene hydrocarbon, labda-7,13(16),14-triene (4).

  15. A new Labdane Diterpene and Other Constituents from Marrubium deserti Noe ex coss.

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    Hocine Dendougui

    2011-01-01

    Full Text Available The phytochemical study of the chloroform soluble part of the hydroalcoholic extract of Marrubium deserti allowed us to describe a new labdane diterpene, 6-dehydroxy-19-acetyl-marrubenol (3 beside three other diterpenes : 19-acetyl-marrubenol (6 , 6-acetyl-marrubenol (7 and 16-epoxy-9-hydroxy-labda-13(16, 14- diene (1. This latter derivative is described for the first time as natural compound. Phytol (2, and three sterols: b -sitosterol (4, stigmasterol (5 and b -sitosterol 3-O-glucoside (8 were also isolated from this species. Structure elucidation of the isolated compounds was accomplished by means of spectroscopic techniques, especially NMR spectroscopy and mass spectrometry.

  16. Labdane-Type Diterpene and Two Flavones from Salvia Sharifii Rech. f. and Esfan. and their Biological Activities

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    Farjam, Mohammad Hossein; Rustaiyan, Abdolhossein; Ezzatzadeh, Elham; Jassbi, Amir Reza

    2013-01-01

    Two flavones, ladanein and 6-hydroxy-5,7,4′-trimethoxyflavone and one labdane-type diterpene, ent-13-epi-manoyloxide, were isolated from an ethyl acetate-methanol extract of the aerial parts of Salvia sharifii. The compounds were purified using several chromatographic methods. Structural elucidation of the compounds was performed using their 1H and 13C-NMR data, EI mass and UV spectral data. The compounds have been subjected to antimicrobial, antioxidant and cytotoxic activity. The diterpene showed higher cytotoxic activity than the flavones while the later compounds were better antioxidants compared with the isolated diterpene. PMID:24250614

  17. Inhibition of human neutrophil elastase by labdane diterpenes from the fruiting bodies of Ramaria formosa.

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    Lee, Ik-Soo; Kim, Kwan-Chul; Yoo, Ick-Dong; Ha, Byung-Jo

    2015-01-01

    Two new labdane diterpenes (1 and 2) were isolated from the fruiting bodies of Ramaria formosa. The structures of these compounds were established by extensive spectroscopic studies and chemical evidence. The inhibitory activity of compounds 1 and 2 against human neutrophil elastase (HNE) was evaluated in vitro. Compounds 1 and 2 inhibited HNE activity moderately. The IC50 values for compounds 1 and 2 were 36.4 ± 1.2 and 40.8 ± 1.5 μM, respectively; the IC50 value for the positive control, EGCG, was 12.5 ± 0.8 μM. In addition, the mechanism by which 2 inhibited HNE was a mixed-type noncompetitive inhibition, with a Ki of 41.5 ± 1.8 μM.

  18. Cancer chemoprevention activity of labdane diterpenes from rhizomes of Hedychium coronarium

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    Denise C. Endringer

    2014-08-01

    Full Text Available Hedychium coronarium J. Koenig, Zingiberaceae, is a medicinal plant popularly used to treat inflammatory conditions in different countries. Three labdane diterpenes [isocoronarin D (1, methoxycoronarin D (2, ethoxycoronarin D (3] and benzoyl eugenol (4 were isolated from rhizomes and their chemopreventive potential was evaluated using in vitro assays, namely the inhibition of NF-κB, COX-1 and -2, the induction of antioxidant response element (ARE, and the inhibition of cell proliferation. Diterpene 1 activated ARE (EC50 57.6 ± 2.4 µM, while 2, 3 and 4 significantly inhibited NF-κB (IC50 of 7.3 ± 0.3, 3.2 ± 0.3 and 32.5 ± 4.9 µM, respectively. In addition, 2 and 3 selectively inhibited COX-1 (IC50 values of 0.9 ± 0.0 and 3.8 ± 0.0 µM, respectively. These data support the potential chemopreventive activity of constituents from H. coronarium rhizomes.

  19. Reconstructing the chemical diversity of labdane-type diterpene biosynthesis in yeast.

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    Ignea, Codruta; Ioannou, Efstathia; Georgantea, Panagiota; Loupassaki, Sofia; Trikka, Fotini A; Kanellis, Angelos K; Makris, Antonios M; Roussis, Vassilios; Kampranis, Sotirios C

    2015-03-01

    Terpenes are a large class of natural products, many of which are used in cosmetics, pharmaceuticals, or biofuels. However, terpene's industrial application is frequently hindered by limited availability of natural sources or low yields of chemical synthesis. In this report, we developed a modular platform based on standardized and exchangeable parts to reproduce and potentially expand the diversity of terpene structures in Saccharomyces cerevisiae. By combining different module-specific parts, we exploited the substrate promiscuity of class I diterpene synthases to produce an array of labdane-type scaffolds. These were subsequently modified by a scaffold decoration module consisting of a mutant library of a promiscuous cytochrome P450 to afford a range of hydroxylated diterpenes. Further P450 protein engineering yielded dedicated and efficient catalysts for specific products. Terpenes produced include precursors of pharmacologically important compounds, molecules that are difficult to obtain from natural sources, or new natural products. The approach described here provides a platform on which additional gene mining, combinatorial biosynthesis, and protein engineering efforts can be integrated to sustainably explore the terpene chemical space.

  20. New Cytotoxic Seco-Type Triterpene and Labdane-Type Diterpenes from Nuxia oppositifolia

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    Shaza M. Al-Massarani

    2017-03-01

    Full Text Available Chromatographic purification of the n-hexane and dichloromethane extracts of Nuxia oppositifolia aerial parts, growing in Saudi Arabia, resulted in the isolation and characterization of three new labdane-type diterpene acids, 2β-acetoxy-labda-7-en-15-oic acid (1, 2β-acetoxy-7-oxolabda-8-en-15-oic acid (2, 2β-acetoxy-6-oxolabda-7-en-15-oic acid (3, and one new seco-triterpene, 3,4-seco olean-12-en-3,30 dioic acid (4, together with 10 known lupane, oleanane and ursane-type triterpenes, as well as the common phytosterols, β-sitosterol and stigmasterol (5–16. Their structures have been assigned on the basis of different spectroscopic techniques including 1D and 2D NMR. Moreover, 13 of the isolated compounds were tested on the human cancer cell lines HeLa (cervical, A549 (lung and MDA (breast, and most of the compounds showed potent cytotoxic activities in vitro.

  1. A New ent-Labdane Diterpene Glycoside form the Leaves of Casearia sylvestris

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    Sylvestin (1), a new ent-labdane glycoside, was isolated from the leaves of Casearia sylvestris. The structure was determined on the basis of 1D and 2 D NMR and HR-ESI-MS analyses. The diterpenoid of ent-labdane type was isolated for the first time from C. sylvestris....

  2. Crystallization and X-ray diffraction analysis of a putative bacterial class I labdane-related diterpene synthase.

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    Serrano-Posada, Hugo; Centeno-Leija, Sara; Rojas-Trejo, Sonia; Stojanoff, Vivian; Rodríguez-Sanoja, Romina; Rudiño-Piñera, Enrique; Sánchez, Sergio

    2015-09-01

    Labdane-related diterpenoids are natural products with potential pharmaceutical applications that are rarely found in bacteria. Here, a putative class I labdane-related diterpene synthase (LrdC) identified by genome mining in a streptomycete was successfully crystallized using the microbatch method. Crystals of the LrdC enzyme were obtained in a holo form with its natural cofactor Mg(2+) (LrdC-Mg(2+)) and in complex with inorganic pyrophosphate (PPi) (LrdC-Mg(2+)-PPi). Crystals of native LrdC-Mg(2+) diffracted to 2.50 Å resolution and belonged to the trigonal space group P3221, with unit-cell parameters a = b = 107.1, c = 89.2 Å. Crystals of the LrdC-Mg(2+)-PPi complex grown in the same conditions as the native enzyme with PEG 8000 diffracted to 2.36 Å resolution and also belonged to the trigonal space group P3221. Crystals of the LrdC-Mg(2+)-PPi complex grown in a second crystallization condition with PEG 3350 diffracted to 2.57 Å resolution and belonged to the monoclinic space group P21, with unit-cell parameters a = 49.9, b = 104.1, c = 66.5 Å, β = 111.4°. The structure was determined by the single-wavelength anomalous dispersion (SAD) technique using the osmium signal from a potassium hexachloroosmate (IV) derivative.

  3. A Labdane diterpene from the aerial parts of Egletes viscosa less

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    Silva-Filho, Francisco Artur e; Lima, Mary Anne S.; Silveira, Edilberto R. [Universidade Federal do Ceara (UFCE), Fortaleza, CE (Brazil). Curso de Pos-Graduacao em Qumica Organica]. E-mail: mary@dqoi.ufc.br; Bezerra, Antonio Marcos E. [Universidade Federal do Ceara (UFCE), Fortaleza, CE (Brazil). Dept. de Fitotecnia; Braz Filho, Raimundo [Universidade Estadual do Norte Fluminense (UENF), Campos dos Goytacases, RJ (Brazil). Setor de Quimica de Produtos Naturais

    2007-07-01

    A new labdane glycoside, 8{alpha}-hydroxylabd-14(15)-ene-13(S)-O-{beta}-D-ribopyranoside, was isolated from the aerial parts of Egletes viscosa Less, along with 13-epi-sclareol, barbatol, tarapacol, spinasterol, ternatin and triacontane. The structures of the isolated compounds were elucidated by the use of spectrometric methods and preparation of chemical derivatives. (author)

  4. Cytotoxic Labdane-type Diterpenes and Diterpene Derivative from Hedychium villosum%毛姜花中的细胞毒活性二萜成分

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    赵庆; 邹澄; 余琴; 赵声定; 何红平; 郝小江

    2012-01-01

    从毛姜花(Hedychium villosum Wall.)中分离得到1个新二萜(1)和1个已知二萜(3).新二萜命名为Villosumcoronarin,该化合物在氯仿或甲醇溶液中发生异构化,部分转化为其差向异构体1’;其平面结构经质谱、红外光谱、氧谱、碳谱及无畸变极化转移增强技术(DEPT)、氢氧相关谱(1H-1H COSY)和异核多键相关谱(HMBC)等方法确证;其相对构型由氢谱、ROESY及量化计算确证.化合物3用琼斯试剂氧化后得到化合物4.生物活性筛选结果表明,化合物1对人白血病细胞株(HL-60)和肝癌细胞株( SMMC-7721)具有强的细胞毒活性,其活性超过阳性对照顺铂(DDP).化合物4对HL-60和SMMC-7721细胞具有显著的细胞毒活性.%Research on active components from rhizomes of Hedychium genus has led to isolation of some diterpenes with significant cytotoxic activities against Chinese hamsler V-79 cells, KB cells, K562 cells, A549 cells, etc. . Hedychium viUosum Wall, is a perennial rhizomatous herb distributed in China and partially in Southeast Asia, which is used in the treatment of pain and inflammation. A new labdane-type diterpene(l), together with a known one(3), were isolated from Hedychium villosum. The new diterpene was named as vil-losumcoronarin, which partially isomerized to its epimer 1' in solution, e. G. , chloroform or methanol. The gross structures of compounds 1 and 1' were elucidated on the basis of MS, IR, 1H NMR, 13C-DEPT NMR, 1H-1H COSY and HMBC. The relative configuration was determined by 1H NMR, ROESY and computational methods. Compound 1 was identified as( 115, 15R)-ll-hydroxy-8( 17) , 12(E)-labdadien-l5 ,16-dial 11,15-hemiacetal. The known compound 3 was oxidized by Jones reagent to get diterpene derivative 4. Bioassays revealed that compound 1 had strong cytotoxic activity against HL-60 and SMMC-7721 cells with IC50 values of 0. 74 and 4. 48μmol/L, respectively, the activities were stronger than the positive control, cis

  5. A copal-8-ol diphosphate synthase from the angiosperm Cistus creticus subsp. creticus is a putative key enzyme for the formation of pharmacologically active, oxygen-containing labdane-type diterpenes.

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    Falara, Vasiliki; Pichersky, Eran; Kanellis, Angelos K

    2010-09-01

    The resin of Cistus creticus subsp. creticus, a plant native to Crete, is rich in labdane-type diterpenes with significant antimicrobial and cytotoxic activities. The full-length cDNA of a putative diterpene synthase was isolated from a C. creticus trichome cDNA library. The deduced amino acid sequence of this protein is highly similar (59%-70% identical) to type B diterpene synthases from other angiosperm species that catalyze a protonation-initiated cyclization. The affinity-purified recombinant Escherichia coli-expressed protein used geranylgeranyl diphosphate as substrate and catalyzed the formation of copal-8-ol diphosphate. This diterpene synthase, therefore, was named CcCLS (for C. creticus copal-8-ol diphosphate synthase). Copal-8-ol diphosphate is likely to be an intermediate in the biosynthesis of the oxygen-containing labdane-type diterpenes that are abundant in the resin of this plant. RNA gel-blot analysis revealed that CcCLS is preferentially expressed in the trichomes, with higher transcript levels found in glands on young leaves than on fully expanded leaves, while CcCLS transcript levels increased after mechanical wounding. Chemical analyses revealed that labdane-type diterpene production followed a similar pattern, with higher concentrations in trichomes of young leaves and increased accumulation upon wounding.

  6. A Copal-8-ol Diphosphate Synthase from the Angiosperm Cistus creticus subsp. creticus Is a Putative Key Enzyme for the Formation of Pharmacologically Active, Oxygen-Containing Labdane-Type Diterpenes1[OA

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    Falara, Vasiliki; Pichersky, Eran; Kanellis, Angelos K.

    2010-01-01

    The resin of Cistus creticus subsp. creticus, a plant native to Crete, is rich in labdane-type diterpenes with significant antimicrobial and cytotoxic activities. The full-length cDNA of a putative diterpene synthase was isolated from a C. creticus trichome cDNA library. The deduced amino acid sequence of this protein is highly similar (59%–70% identical) to type B diterpene synthases from other angiosperm species that catalyze a protonation-initiated cyclization. The affinity-purified recombinant Escherichia coli-expressed protein used geranylgeranyl diphosphate as substrate and catalyzed the formation of copal-8-ol diphosphate. This diterpene synthase, therefore, was named CcCLS (for C. creticus copal-8-ol diphosphate synthase). Copal-8-ol diphosphate is likely to be an intermediate in the biosynthesis of the oxygen-containing labdane-type diterpenes that are abundant in the resin of this plant. RNA gel-blot analysis revealed that CcCLS is preferentially expressed in the trichomes, with higher transcript levels found in glands on young leaves than on fully expanded leaves, while CcCLS transcript levels increased after mechanical wounding. Chemical analyses revealed that labdane-type diterpene production followed a similar pattern, with higher concentrations in trichomes of young leaves and increased accumulation upon wounding. PMID:20595348

  7. Genus Cistus: a model for exploring labdane-type diterpenes' biosynthesis and a natural source of high value products with biological, aromatic and pharmacological properties

    Science.gov (United States)

    Papaefthimiou, Dimitra; Papanikolaou, Antigoni; Falara, Vasiliki; Givanoudi, Stella; Kostas, Stefanos; Kanellis, Angelos

    2014-06-01

    The family Cistaceae (Angiosperm, Malvales) consists of 8 genera and 180 species, with 5 genera native of the Mediterranean area (Cistus, Fumara, Halimium, Helianthemum and Tuberaria). Traditionally, a number of Cistus specie have been used in Mediterranean folk medicine as herbal tea infusions for healing, digestive problems and colds, as extracts for the treatment of diseases, and as fragrances. The resin, ladano, secreted by the glandular trichomes of certain Cistus species contains a number of phytochemicals with antioxidant, antibacterial, antifungal and anticancer properties. Furthermore, total leaf aqueous extracts possess anti-influenza virus activity. All these properties have been attributed to phytochemicals such as terpenoids, including diterpenes, labdane-type diterpenes and clerodanes, phenylpropanoids, including flavonoids and ellagitannins, several groups of alkaloids and other types of secondary metabolites. In the past 20 years, research on Cistus involved chemical, biological and phylogenetic analysis but recent investigations have involved genomic and molecular approaches. Our lab is exploring the biosynthetic machinery that generates terpenoids and phenylpropanoids, with a goal to harness their numerous properties that have applications in the pharmaceutical, chemical and aromatic industries. This review focuses on the systematics, botanical characteristics, geographic distribution, chemical analyses, biological function and biosynthesis of major compounds, as well as genomic analyses and biotechnological approaches of the main Cistus species found in the Mediterranean basin, namely C. albidus, C. creticus, C. crispus, C. parviflorus, C. monspeliensis, C. populifolius, C. salviifolius, C. ladanifer, C. laurifolius and C. clusii.

  8. Genus Cistus: a model for exploring labdane-type diterpenes' biosynthesis and a natural source of high value products with biological, aromatic and pharmacological properties

    Directory of Open Access Journals (Sweden)

    Dimitra ePapaefthimiou

    2014-06-01

    Full Text Available The family Cistaceae (Angiosperm, Malvales consists of 8 genera and 180 species, with 5 genera native of the Mediterranean area (Cistus, Fumara, Halimium, Helianthemum and Tuberaria. Traditionally, a number of Cistus specie have been used in Mediterranean folk medicine as herbal tea infusions for healing, digestive problems and colds, as extracts for the treatment of diseases, and as fragrances. The resin, ladano, secreted by the glandular trichomes of certain Cistus species contains a number of phytochemicals with antioxidant, antibacterial, antifungal and anticancer properties. Furthermore, total leaf aqueous extracts possess anti-influenza virus activity. All these properties have been attributed to phytochemicals such as terpenoids, including diterpenes, labdane-type diterpenes and clerodanes, phenylpropanoids, including flavonoids and ellagitannins, several groups of alkaloids and other types of secondary metabolites. In the past 20 years, research on Cistus involved chemical, biological and phylogenetic analysis but recent investigations have involved genomic and molecular approaches. Our lab is exploring the biosynthetic machinery that generates terpenoids and phenylpropanoids, with a goal to harness their numerous properties that have applications in the pharmaceutical, chemical and aromatic industries. This review focuses on the systematics, botanical characteristics, geographic distribution, chemical analyses, biological function and biosynthesis of major compounds, as well as genomic analyses and biotechnological approaches of the main Cistus species found in the Mediterranean basin, namely C. albidus, C. creticus, C. crispus, C. parviflorus, C. monspeliensis, C. populifolius, C. salviifolius, C. ladanifer, C. laurifolius and C. clusii.

  9. A labdane diterpene exerts ex vivo and in vivo cardioprotection against post-ischemic injury: involvement of AKT-dependent mechanisms.

    Science.gov (United States)

    Cuadrado-Berrocal, Irene; Gómez-Gaviro, María V; Benito, Yolanda; Barrio, Alicia; Bermejo, Javier; Fernández-Santos, María Eugenia; Sánchez, Pedro L; Desco, Manuel; Fernández-Avilés, Francisco; Fernández-Velasco, María; Boscá, Lisardo; de Las Heras, Beatriz

    2015-02-15

    Therapeutic approaches to protect the heart from ischemia/reperfusion (I/R) injury are an area of intense research, as myocardial infarction is a major cause of mortality and morbidity. Diterpenes are bioactive natural products with great therapeutic potential. In the present study, we have investigated the in vivo cardioprotective effects of a labdane diterpene (DT1) against cardiac I/R injury and the molecular mechanisms involved. DT1 attenuates post-ischemic injury via an AKT-dependent activation of HIF-1α, survival pathways and inhibition of NF-κB signaling. Myocardial infarction (MI) was induced in Wistar rats occluding the left coronary artery (LCA) for 30min followed by 72h reperfusion. DT1 (5mg/kg) was intravenously administered at reperfusion. In addition, we investigated the mechanisms of cardioprotection in the Langendorff-perfused model. Cardioprotection was observed when DT1 was administered after myocardial injury. The molecular mechanisms involved the activation of the survival pathway PDK-1, AKT and AMPK, a reduced phosphorylation of PKD1/2 and sustained HIF-1α activity, leading to increased expression of anti-apoptotic proteins and decreased caspase-3 activation. Pharmacological inhibition of AKT following MI and prior to DT1 challenge significantly decreased the cardioprotection afforded by DT1 therapy at reperfusion. Cardiac function after MI was significantly improved after DT1-treatment, as evidenced by hemodynamic recovery and decreased myocardial infarct size. These findings demonstrate an efficient in vivo cardioprotection by diterpene DT1 against I/R when administered at reperfusion, opening new therapeutic strategies as adjunctive therapy for the pharmacological management of I/R injury.

  10. Three new labdane-type diterpene glycosides from fruits of Rubus chingii and their cytotoxic activities against five humor cell lines.

    Science.gov (United States)

    Zhong, Ruijian; Guo, Qing; Zhou, Guoping; Fu, Huizheng; Wan, Kaihua

    2015-04-01

    Three new labdane-type diterpene glycosides, 15,18-di-O-β-d-glucopyranosyl-13(E)-ent-labda-7(8),13(14)-diene-3β,15,18-triol (1), 15,18-di-O-β-d-glucopyranosyl-13(E)-ent-labda-8(9),13(14)-diene-3β,15,18-triol (2), and 15-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranosyl-18-O-β-d-glucopyranosyl-13(E)-ent-labda-8(9),13(14)-diene-3β,15,18-triol (3), were isolated from the fruits of Rubus chingii. Their structures were elucidated on the basis of spectroscopic data and chemical methods. The cytotoxic activities of compounds 1-3 were evaluated against five human tumor cell lines (HCT-8, BGC-823, A549, and A2780). Compounds 3 showed cytotoxic activity against A549 with an IC50 value of 2.32μM.

  11. FT-IR, FT-Raman, UV-visible, and NMR spectroscopy and vibrational properties of the labdane-type diterpene 13-epi-sclareol.

    Science.gov (United States)

    Chain, Fernando E; Leyton, Patricio; Paipa, Carolina; Fortuna, Mario; Brandán, Silvia A

    2015-03-05

    In this work, FT-IR, FT-Raman, UV-Visible and NMR spectroscopies and density functional theory (DFT) calculations were employed to study the structural and vibrational properties of the labdane-type diterpene 13-epi-sclareol using the hybrid B3LYP method together with the 6-31G(∗) basis set. Three stable structures with minimum energy found on the potential energy curves (PES) were optimized, and the corresponding molecular electrostatic potentials, atomic charges, bond orders, stabilization energies and topological properties were computed at the same approximation level. The complete assignment of the bands observed in the vibrational spectrum of 13-epi-sclareol was performed taking into account the internal symmetry coordinates for the three structures using the scaled quantum mechanical force field (SQMFF) methodology at the same level of theory. In addition, the force constants were calculated and compared with those reported in the literature for similar compounds. The predicted vibrational spectrum and the calculated (1)H NMR and (13)C NMR chemical shifts are in good agreement with the corresponding experimental results. The theoretical UV-Vis spectra for the most stable structure of 13-epi-sclareol demonstrate a better correlation with the corresponding experimental spectrum. The study of the three conformers by means of the theory of atoms in molecules (AIM) revealed different H bond interactions and a strong dependence of the interactions on the distance between the involved atoms. Furthermore, the natural bond orbital (NBO) calculations showed the characteristics of the electronic delocalization for the two six-membered rings with chair conformations.

  12. FT-IR, FT-Raman, UV-Visible, and NMR spectroscopy and vibrational properties of the labdane-type diterpene 13-epi-sclareol

    Science.gov (United States)

    Chain, Fernando E.; Leyton, Patricio; Paipa, Carolina; Fortuna, Mario; Brandán, Silvia A.

    2015-03-01

    In this work, FT-IR, FT-Raman, UV-Visible and NMR spectroscopies and density functional theory (DFT) calculations were employed to study the structural and vibrational properties of the labdane-type diterpene 13-epi-sclareol using the hybrid B3LYP method together with the 6-31G∗ basis set. Three stable structures with minimum energy found on the potential energy curves (PES) were optimized, and the corresponding molecular electrostatic potentials, atomic charges, bond orders, stabilization energies and topological properties were computed at the same approximation level. The complete assignment of the bands observed in the vibrational spectrum of 13-epi-sclareol was performed taking into account the internal symmetry coordinates for the three structures using the scaled quantum mechanical force field (SQMFF) methodology at the same level of theory. In addition, the force constants were calculated and compared with those reported in the literature for similar compounds. The predicted vibrational spectrum and the calculated 1H NMR and 13C NMR chemical shifts are in good agreement with the corresponding experimental results. The theoretical UV-Vis spectra for the most stable structure of 13-epi-sclareol demonstrate a better correlation with the corresponding experimental spectrum. The study of the three conformers by means of the theory of atoms in molecules (AIM) revealed different H bond interactions and a strong dependence of the interactions on the distance between the involved atoms. Furthermore, the natural bond orbital (NBO) calculations showed the characteristics of the electronic delocalization for the two six-membered rings with chair conformations.

  13. O Ácido (817,12E,14-labdatrieno-18-óico (labdano302, diterpeno tipo labdano isolado de Xylopia langsdorffiana St. Hil. & Tul. (Annonaceae relaxa a traquéia isolada de cobaia The (817,12E,14-labdatrien-18-oic acid (labdane302, labdane-type diterpene isolated from Xylopia langsdorffiana St. Hil. & Tul. (Annonaceae relaxes the guinea-pig trachea

    Directory of Open Access Journals (Sweden)

    Luciano A. A. Ribeiro

    2007-06-01

    Full Text Available A espécie Xylopia langsdorffiana St. Hil.. & Tul. é popularmente conhecida como "pimenteira-da-terra" no Sudeste do Brasil. A partir do fracionamento do extrato etanólico, obtido das cascas do caule desta espécie, foi isolado um diterpeno tipo labdano, identificado como sendo o ácido 8(17,12E,14-labdatrieno-18-óico, e que neste trabalho é codificado como labdano302. O labdano302 relaxou o tônus basal dos anéis de traquéia isolada de cobaia com um valor de CE50 de 6,7 ± 0,5 x 10-8 M. O diterpeno labdano302 relaxou de maneira dependente de concentração os anéis pré-contraídos com carbacol (10-6 M, tanto na presença (CE50 = 1,4 ± 0,7 x 10-5 M como na ausência de epitélio funcional (CE50 = 1,5 ± 0,3 x 10-5 M, bem como anéis pré-contraídos com 18 ou 60 mM de KCl, apresentando valores de CE50 de 2,3 ± 0,4 x 10(7 M e 1,8 ± 0,8 x 10-5 M, respectivamente. Este efeito relaxante, sobre as contrações induzidas por 18 mM de KCl, tanto foi significantemente mais potente quanto mais eficaz quando comparado ao efeito sobre as contrações induzidas por 60 mM de KCl. Assim, labdano302 mostra um efeito relaxante em traquéia isolada de cobaia, tanto em seu tônus basal como sob estímulo contrátil, aparentemente sem a participação dos fatores relaxantes derivados do epitélio, contudo com possível participação dos canais de K+.Xylopia langsdorfiana St. Hil. & Tul. is popularly known as "pimenteira-da-terra" in Southeast of Brazil. The fractionation of the ethanol extract obtained from the stem-bark of this species yielded a labdane-type diterpene identified as 8(17,12E,14-labdatrien-18-oic acid, referred here as labdane302. In this study, we investigated the effect of labdane302 in guinea-pig trachea. labdane302 relaxed the basal tonus of trachea rings with EC50 value of 6.7 ± 0.5 x 10-8 M. The diterpene labdane302 relaxed the pre-contracted rings by carbachol 10-6 M both in the presence (EC50 = 1.4 ± 0.7 x 10-5 M and absence

  14. Resin Diterpenes from Austrocedrus chilensis

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    Verónica Rachel Olate

    2011-12-01

    Full Text Available Seventeen diterpenes belonging to the labdane, abietane and isopimarane skeleton classes were isolated from the resin of the Chilean gymnosperm Austrocedrus chilensis and identified by spectroscopic and spectrometric methods. The diterpene 12-oxo-labda-8(17,13E-dien-19 oic acid is reported for the first time as a natural product and 14 diterpenes are reported for the first time for the species.

  15. Synthesis of Labdane Natural Product-like Diterpenes%Labdane二萜“类天然产物”的合成研究

    Institute of Scientific and Technical Information of China (English)

    张军庆; 刘跃佳; 宁弘历; 廖盼; 杨海君

    2015-01-01

    以香紫苏内酯为原料,通过内酯开环、Kishner-Wolff-Huang 还原等反应合成得到6个含有不同芳环的lab-dane二萜“类天然产物”1(a-c)和2(a-c),所得化合物经红外、质谱、1H NMR等表征。%Six labdane-like compounds 1( a-c)and 2( a-c)with different aryl groups were synthe-sized through lactone ring opening and Kishner-Wolff-Huang reduction starting from sclareolide. All the compounds obtained were characterized by IR,MS,and 1 H NMR.

  16. Biosynthesis of mercapturic acid derivative of the labdane-type diterpene, cyslabdan that potentiates imipenem activity against methicillin-resistant Staphylococcus aureus: cyslabdan is generated by mycothiol-mediated xenobiotic detoxification.

    Science.gov (United States)

    Ikeda, Haruo; Shin-Ya, Kazuo; Nagamitsu, Tohru; Tomoda, Hiroshi

    2016-03-01

    Genome mining of cyslabdan-producing Streptomyces cyslabdanicus K04-0144 revealed that a set of four genes, cldA, cldB, cldC, and cldD (the cld cluster), which formed a single transcriptional unit, were involved in the biosynthesis of cyslabdan that potentiates imipenem activity against methicillin-resistant Staphylococcus aureus. Experimental studies supported the heterologous expression of the cld cluster of S. cyslabdanicus K04-0144 in S. avermitilis SUKA22, and transformants carrying the cld cluster produced not only cyslabdan A (1), but also its new derivatives, 17-hydroxyl-1 (2) and 2-hydroxyl-1 (3), in the culture broth. An analysis of diterpene metabolites in the mycelia showed that a large amount of a novel intermediate had accumulated and its structure was elucidated as (7S, 8S, 12E)-8,17-epoxy-7-hydroxylabda-12,14-diene (4). The cld-like cluster (rmn cluster) was also detected in the genome of S. anulatus GM95 by searching our in-house genome databases, and the heterologous expression of the rmn cluster in S. avermitilis SUAK22 demonstrated that the rmn cluster was involved in the biosynthesis of the labdane-type bicyclic diterpene, raimonol (7). CldA/RmnA catalyzed the generation of geranylgeranyl diphosphate (GGPP) from dimethylallyl diphosphate and isopentenyl diphosphate. CldB/RmnB converted GGPP to (+)-copalyl diphosphate, and CldD/RmnD generated labda-8(17),12(E),14-triene (5). CldC introduced two oxygen atoms at C-7 and C-8,17 to generate 4, while RmnC hydroxylated 5 at C-7 to generate 7. The heterologous expression of the cld cluster suggested that four gene products catalyzed to generate 4, but not 1. The deletion mutant of the gene encoding the mycothiol (MSH)-S-conjugate amidase (mca) of S. avermitilis SUKA22 carrying the cld cluster failed to produce 1, but accumulated 4 in the mycelia, whereas S. avermitilis SUKA22 and its mca-deletion mutant carrying the cld cluster both produced the MSH-S-conjugate of 4. The intermediate 4 was converted

  17. Isoleojaponin, a new Halimane diterpene isolated from Leonurus japonicus.

    Science.gov (United States)

    Wu, Hankui; Wang, Sensheng; Xu, Zhiyong; Sun, Shanshan; Liu, Haijuan; Wang, Jinjin; E, Yan; Lv, Yanyan; Dong, Xuelin; Li, Gang; Zhang, Liping; Shi, Yunfeng

    2015-01-07

    Leojaponin (2), a labdane diterpene, was isolated from the EtOH extract of the herb of Leonurus japonicus together with a new halimane diterpene named isoleojaponin (1). Isoleojaponin has a new diterpene skeleton with a unique cross-conjugated α,β-unsaturated ketone system, Their structures were elucidated by physical and spectroscopic analysis, and the relative configuration of the chiral C-9 carbon was determined by a computational method, and analysis of its possible biogenesis pathways.

  18. Isoleojaponin, a New Halimane Diterpene Isolated from Leonurus japonicus

    Directory of Open Access Journals (Sweden)

    Hankui Wu

    2015-01-01

    Full Text Available Leojaponin (2, a labdane diterpene, was isolated from the EtOH extract of the herb of Leonurus japonicus together with a new halimane diterpene named isoleojaponin (1. Isoleojaponin has a new diterpene skeleton with a unique cross-conjugated α,β-unsaturated ketone system, Their structures were elucidated by physical and spectroscopic analysis, and the relative configuration of the chiral C-9 carbon was determined by a computational method, and analysis of its possible biogenesis pathways.

  19. Diterpenes from Coleus forskohlii (WILLD.) BRIQ. (Labiatae).

    Science.gov (United States)

    Shan, Yupei; Xu, Lingling; Lu, Yang; Wang, Xiaobing; Zheng, Qitai; Kong, Lingyi; Niwa, Masatake

    2008-01-01

    Three new minor labdane diterpene glycosides, forskoditerpenoside C, D, and E (1-3), and a novel labdane diterpene forskoditerpene A (4) were isolated from the ethanol extract of the whole plant of Coleus forskohlii. Their structures and relative stereochemistry were elucidated on the basis of extensive spectroscopic analyses including 1D-, 2D-NMR, and HR-ESI-MS experiments. Compounds 1-3 showed an unusual 8,13-epoxy-labd-14-en-11-one glycoside pattern. They showed relaxative effects on isolated guinea pig tracheal spirals in vitro. Compound 4 with a cyclopropyl, confirmed by a single-crystal X-ray diffraction determination, is the first known labdane derivative with a spiro element.

  20. Establishment of callus, cell suspension and shoot cultures of Leonurus cardiaca L. and diterpene analysis.

    Science.gov (United States)

    Knöss, W

    1995-10-01

    Callus cultures, cell suspension cultures and shoot cultures of Leonurus cardiaca L. (Motherwort) were established and growth conditions optimized. Shoot cultures showed constant growth whether in the dark or under continuous light, accumulating varying amounts of the furanic labdane diterpenes leosibiricin, preleosibirin, leosibirin and isoballotenol acetate, which are also present in the soil-grown plants. Only traces of leosibiricin were detected in callus cultures, while cell suspension cultures did not produce any furanic diterpenes. A small amount of furanic labdane diterpenes was found in the medium of shoot cultures. Callus and shoot culture induction of several other Lamiaceae species is also described.

  1. Two minor diterpene glycosides and an eudesman sesquiterpene from Coleus forskohlii.

    Science.gov (United States)

    Shan, Yupei; Wang, Xiaobing; Zhou, Xiang; Kong, Lingyi; Niwa, Masatake

    2007-03-01

    Two new labdane diterpene glycosides, forskoditerpenosides A, B (1, 2) and a new eudesmane sesquiterpene, 4beta,7beta,11-enantioeudesmantriol (3), were isolated from the ethanol extract of the whole plant of Coleus forskohlii. Their structures and stereochemistry were elucidated by extensive spectroscopic analysis and chemical methods. The structure of compound 3 was confirmed by X-ray diffraction. This is the first report about the occurrence of glycosides derived from the kind of labdane diterpene, 8,13-epoxy-labd-14-en-11-one, in the nature. Compounds 1 and 2 showed relaxative effects on isolated guinea pig tracheal spirals in vitro.

  2. Labdane-type diterpenoids from hairy root cultures of Coleus forskohlii, possible intermediates in the biosynthesis of forskolin.

    Science.gov (United States)

    Asada, Yoshihisa; Li, Wei; Terada, Tomohiro; Kuang, Xinzhu; Li, Qin; Yoshikawa, Takafumi; Hamaguchi, Shogo; Namekata, Iyuki; Tanaka, Hikaru; Koike, Kazuo

    2012-07-01

    Significant attention has been devoted to studying hairy root cultures as a promising strategy for production of various valuable secondary metabolites. These offer many advantages, such as high growth rate, genetic stability and being hormone-free. In this study, a detailed phytochemical investigation of the secondary metabolites of Coleus forskohlii hairy root cultures was undertaken and which resulted in the isolation of 22 compounds, including four forskolin derivatives and a monoterpene. Their structures were elucidated by extensive spectroscopic analyses. These compounds could be classified into four groups viz.: labdane-type diterpenes, monoterpenes, triterpenes and phenylpropanoid dimers. Apart from one compound, all labdane type diterpenes are oxygenated at C-11 as in forskolin and a scheme showing their biosynthetic relationships is proposed.

  3. Synthesis of labdane diterpenes galanal A and B from (+)-sclareolide.

    Science.gov (United States)

    Kumar, Chebolu Naga Sesha Sai Pavan; Chein, Rong-Jie

    2014-06-06

    The first chemical synthesis of galanal A and B was achieved by a concise and highly efficient pathway starting from commercially available (+)-sclareolide and features a Wittig reaction and a titanocene-mediated radical cyclization as the key steps.

  4. Chlorosilphanol A and silphanepoxol, labdane diterpenes from Silphium perfoliatum.

    Science.gov (United States)

    Pcolinski, M J; Doskotch, R W; Lee, A Y; Clardy, J

    1994-06-01

    Two carterochaetols, chlorosilphanol A and silphanepoxol, isolated from the leaves of Silphium perfoliatum were assigned structures 1 and 3, respectively, from spectral studies and with absolute stereochemistry established by X-ray crystallography of 1. Complete 1H- and 13C-nmr assignments for the parent compounds and their acetates 2 and 4 were made at high-field using 1D and 2D methods, and chlorosilphanol A was chemically converted via its acetate to silphanepoxol. Revision of structure for related carterochaetols in the literature is required, in particular with respect to the stereochemistry at C-12 and that of the side-chain.

  5. Antileishmanial activity of natural diterpenes from Cistus sp. and semisynthetic derivatives thereof.

    Science.gov (United States)

    Fokialakis, Nikolas; Kalpoutzakis, Eleftherios; Tekwani, Babu Lal; Skaltsounis, Alexios Leandros; Duke, Stephen Oscar

    2006-08-01

    Eleven cis-clerodane diterpenes, seven labdane type diterpenes and one triterpene isolated from Cistus monspeliensis and the resin "Ladano" of Cistus creticus subsp. creticus were evaluated against Leishmania donovani promastigotes, the causative agent for visceral leishmaniasis. In addition, eleven semisynthetic manoyl oxide, seventeen labdane type derivatives and a triterpene were also evaluated for their antileishmanial activity. 18-Acetoxy-cis-clerod-3-en-15-ol, 15,18-diacetoxy-cis-clerod-3-ene and 13-(E)-8a-hydroxylabd-13-en-15-ol 2-chloroethylcarbamate exhibited the most potent and selective leishmanicidal activity with IC50 values of 3.3 microg/ml, 3.4 microg/ml and 3.5 microg/ml, respectively.

  6. Labdane Diterpenoids from Coleus forskohlii (Willd.) Briq.

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    In order to investigate the active substances of Coleus forskohlii (Willd.) Briq., a new labdane diterpenoid, 12-hydroxy-8,13E-labdadien-15-0ic acid, together with two 8,13-epoxylabd-14-ene-11-one diterpenoids, 6-acetyl-1-deoxyforskolin and 6-acetyl-1,9-dideoxyforskolin, was isolated on the basis of various chromatographic techniques and spectroscopic methods.

  7. The Labdane Ent-3-Acetoxy-Labda-8(17), 13-Dien-15-Oic Decreases Blood Pressure In Hypertensive Rats

    Science.gov (United States)

    Simplicio, Janaina A.; Simão, Marilia R.; Ambrosio, Sergio R.; Tirapelli, Carlos R.

    2016-01-01

    Background Labdane-type diterpenes induce lower blood pressure via relaxation of vascular smooth muscle; however, there are no studies describing the effects of labdanes in hypertensive rats. Objective The present study was designed to investigate the cardiovascular actions of the labdane-type diterpene ent-3-acetoxy-labda-8(17), 13-dien-15-oic acid (labda-15-oic acid) in two-kidney 1 clip (2K-1C) renal hypertension. Methods Vascular reactivity experiments were performed in aortic rings isolated from 2K-1C and normotensive (2K) male Wistar rats. Nitrate/nitrite (NOx) measurement was performed in aortas by colorimetric assay. Blood pressure measurements were performed in conscious rats. Results Labda-15-oic acid (0.1-300 µmol/l) and forskolin (0.1 nmol/l - 1 µmol/l) relaxed endothelium-intact and endothelium-denuded aortas from both 2K-1C and 2K rats. Labda-15-oic acid was more effective at inducing relaxation in endothelium-intact aortas from 2K pre-contracted with phenylephrine when compared to the endothelium-denuded ones. Forskolin was more potent than labda-15-oic acid at inducing vascular relaxation in arteries from both 2K and 2K-1C rats. Labda-15-oic acid-induced increase in NOx levels was lower in arteries from 2K-1C rats when compared to 2K rats. Intravenous administration of labda-15-oic acid (0.3-3 mg/kg) or forskolin (0.1-1 mg/kg) induced hypotension in conscious 2K-1C and 2K rats. Conclusion The present findings show that labda-15-oic acid induces vascular relaxation and hypotension in hypertensive rats. PMID:27096521

  8. The Labdane Ent-3-Acetoxy-Labda-8(17), 13-Dien-15-Oic Decreases Blood Pressure In Hypertensive Rats

    Energy Technology Data Exchange (ETDEWEB)

    Simplicio, Janaina A. [Programa de Pós-Graduação em Farmacologia - Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo (USP), Ribeirão Preto, SP (Brazil); Departamento de Enfermagem Psiquiátrica e Ciências Humanas - Laboratório de Farmacologia - Escola de Enfermagem de Ribeirão Preto (USP), Ribeirão Preto, SP (Brazil); Simão, Marilia R.; Ambrosio, Sergio R. [Núcleo de Pesquisa em Ciências e Tecnologia - Universidade de Franca (UNIFRAN), Franca, SP (Brazil); Tirapelli, Carlos R., E-mail: crtirapelli@eerp.usp.br [Departamento de Enfermagem Psiquiátrica e Ciências Humanas - Laboratório de Farmacologia - Escola de Enfermagem de Ribeirão Preto (USP), Ribeirão Preto, SP (Brazil)

    2016-06-15

    Labdane-type diterpenes induce lower blood pressure via relaxation of vascular smooth muscle; however, there are no studies describing the effects of labdanes in hypertensive rats. The present study was designed to investigate the cardiovascular actions of the labdane-type diterpene ent-3-acetoxy-labda-8(17), 13-dien-15-oic acid (labda-15-oic acid) in two-kidney 1 clip (2K-1C) renal hypertension. Vascular reactivity experiments were performed in aortic rings isolated from 2K-1C and normotensive (2K) male Wistar rats. Nitrate/nitrite (NOx) measurement was performed in aortas by colorimetric assay. Blood pressure measurements were performed in conscious rats. Labda-15-oic acid (0.1-300 µmol/l) and forskolin (0.1 nmol/l - 1 µmol/l) relaxed endothelium-intact and endothelium-denuded aortas from both 2K-1C and 2K rats. Labda-15-oic acid was more effective at inducing relaxation in endothelium-intact aortas from 2K pre-contracted with phenylephrine when compared to the endothelium-denuded ones. Forskolin was more potent than labda-15-oic acid at inducing vascular relaxation in arteries from both 2K and 2K-1C rats. Labda-15-oic acid-induced increase in NOx levels was lower in arteries from 2K-1C rats when compared to 2K rats. Intravenous administration of labda-15-oic acid (0.3-3 mg/kg) or forskolin (0.1-1 mg/kg) induced hypotension in conscious 2K-1C and 2K rats. The present findings show that labda-15-oic acid induces vascular relaxation and hypotension in hypertensive rats.

  9. A chiral 1,3,2-dioxaborolane derived from a natural diterpene for asymmetric reduction of prochiral ketones

    Directory of Open Access Journals (Sweden)

    Diego A. Cifuente

    2012-01-01

    Full Text Available The application of a chiral 1,3,2-dioxaborolane, for the sulfide complex-mediated asymmetric reduction of prochiral ketones at room temperature, are described. The B-methoxy-dioxaborolane was synthesized from 2 a ,3 a -dihydroxycativic acid, a labdane-type diterpene isolated from aerial parts of Baccharis scandens DC. Very good chemical yields (85-97% and high enantioselectivities (62-96% ee, were obtained.

  10. Antiproliferative Diterpenes from a Malleastrum sp. from the Madagascar dry forest.

    Science.gov (United States)

    Liu, Yixi; Wiedle, C Houston; Brodie, Peggy J; Callmander, Martin W; Rakotondrajaona, R; Rakotobe, Etienne; Rasamison, Vincent E; Kingston, David G I

    2015-09-01

    An ethanol extract of leaves of the plant species Malleastrum sp. collected in northern Madagascar afforded the new clerodane diterpene 18-oxo-cleroda-3,13-dien-16,15-olide (1), together with the three known clerodane diterpenes 16,18-dihydroxykolavenic acid lactone (2), solidagolactone (3) and (-)-kolavenol (4), and the known labdane diterpene 3-oxo-ent-Iabda-8(17),13-dien-15,16-olide (5). Compounds 1, 3, and 4 showed moderate antiproliferative activities against the A2780 ovarian cancer cell line, with the IC50 values of 3.01 ± 0.8, 7.84 ± 0.2, and 17.9 ± 3 µM, respectively. The structure elucidations of all compounds were carried out based on analysis of NMR and mass spectroscopic data. The relative stereochemistry of compound 1 was determined by NOESY NMR spectrum.

  11. Methods for producing diterpenes

    DEFF Research Database (Denmark)

    2015-01-01

    The present invention discloses that by combining different di TPS enzymes of class I and class II different diterpenes may be produced including diterpenes not identified in nature. Surprisingly it is revealed that a di TPS enzyme of class I of one species may be combined with a di TPS enzyme...... of class II from a different species, resulting in a high diversity of diterpenes, which can be produced....

  12. Alterações ultraestruturais em larvas de Aedes aegypti submetidas ao diterpeno labdano, isolado de Copaifera reticulata (Leguminosae, e à uma fração rica em taninos de Magonia pubescens (Sapindaceae Ultrastructural alterations in larvae of Aedes aegypti subject to labdane diterpene isolated from Copaifera reticulata (Leguminosae and a fraction enriched with tannins of Magonia pubescens (Sapindaceae

    Directory of Open Access Journals (Sweden)

    Cleyde Ferreira Barreto Valotto

    2011-04-01

    Full Text Available INTRODUÇÃO: Dengue é um importante problema de saúde pública, em vários países, e tem como principal vetor o Aedes aegypti, mosquito mais adaptado às áreas urbanizadas. Apresenta-se, pela primeira vez, as alterações ultraestruturais em larvas de 3º estádio, desse mosquito, causadas pelos larvicidas naturais, um diterpeno labdano, extraído de Copaifera reticulata, e uma fração rica em taninos catéquicos, extraída de Magonia pubescens, evidenciando o mecanismo de ação dessas substâncias. MÉTODOS: Os experimentos foram realizados com larvas de 3º estádio em solução de 0,9ppm, do diterpeno (3-β-acetoxylabdan-8(17-13-dien-15-óico e de 3,7ppm, da fração majoritária de tanino catéquico de massa molecular 864Da. Obtiveram-se as substâncias através de fracionamentos cromatográficos sucessivos, identificadas por ressonância magnética nuclear de hidrogênio e espectrometria de massas. As larvas que atingiram estado letárgico foram coletadas e dissecadas e seus tubos digestórios fixados, desidratados, emblocados e polimerizados. Cortes ultrafinos foram feitos e contrastados com acetato de uranila 3% e citrato de chumbo, posteriormente, levados ao microscópio eletrônico. RESULTADOS: As principais alterações ultraestruturais provocadas pelos diterpeno e tanino sobre larvas de Aedes aegypti foram vacuolização citoplasmática, desorganização e degeneração celular, mudança estrutural dos microvilos e deslocamento das células da lâmina basal. CONCLUSÕES: O diterpeno e a fração rica em taninos catéquicos provocaram a morte das larvas de Aedes aegypti através da destruição celular no intestino médio.INTRODUCTION: Dengue is an important public health problem in many countries and its main vector Aedes aegypti, is the mosquito most adapted to urban areas. For the first time, the mechanism of action of labdane diterpenoid extracted from Copaifera reticulata and the fraction enriched of catechin tannins

  13. Phytochemical profiling of Curcuma kwangsiensis rhizome extract, and identification of labdane diterpenoids as positive GABAA receptor modulators.

    Science.gov (United States)

    Schramm, Anja; Ebrahimi, Samad Nejad; Raith, Melanie; Zaugg, Janine; Rueda, Diana C; Hering, Steffen; Hamburger, Matthias

    2013-12-01

    An ethyl acetate extract of Curcuma kwangsiensis S.G. Lee & C.F. Liang (Zingiberaceae) rhizomes (100 μg/ml) enhanced the GABA-induced chloride current (IGABA) through GABAA receptors of the α1β2γ2S subtype by 79.0±7.0%. Potentiation of IGABA was measured using the two-microelectrode voltage-clamp technique and Xenopus laevis oocytes. HPLC-based activity profiling of the crude extract led to the identification of 11 structurally related labdane diterpenoids, including four new compounds. Structure elucidation was achieved by comprehensive analysis of on-line (LC-PDA-ESI-TOF-MS) and off-line (microprobe 1D and 2D NMR) spectroscopic data. The absolute configuration of the compounds was established by comparison of experimental and calculated ECD spectra. Labdane diterpenes represent a new class of plant secondary metabolites eliciting positive GABAA receptor modulation. The highest efficiency was observed for zerumin A (maximum potentiation of IGABA by 309.4±35.6%, and EC50 of 24.9±8.8 μM).

  14. Diterpene glycosides from the ethanol extract of Diplopterygium rufopilosum.

    Science.gov (United States)

    Hu, Jiang; Song, Yan; Li, Hui; Mao, Xia; Shi, Xiao-Dong

    2015-01-01

    A phytochemical investigation on the 70% EtOH extract from the dry fronds of Diplopterygium rufopilosum afforded two new labdane-type diterpene glycosides, (3β,13S)-3-O-β-d-glucopyranosyl-13-O-α-l-rhamnopyranosyl-labda-8(17),14-diene (1) and (3β,13S)-3-O-α-l-rhamnopyranosyl-13-O-[6-O-acetyl-β-d-glucopyranosyl-(1 → 4)-2-O-acetyl-α-l-rhamnopyranosyl]-labda-8(17),14-diene (2). Their structures were determined on the basis of chemical method and spectroscopic analyses, including 1D and 2D NMR (COSY, HMQC, HMBC, and NOESY) and HR-ESI-MS analyses.

  15. Aspects of tobacco diterpene biosynthesis and accumulation

    Energy Technology Data Exchange (ETDEWEB)

    Keene, C.K.

    1985-01-01

    Lamina, midveins, stalks and flowers of most Nicotiana species are covered with trichomes. The exudate which accumulates around glandular trichome heads has been suggested to be responsible for the characteristics flavor and aroma associated with different tobaccos. Many classes of compounds have been identified in cuticular surface washes and exudates of tobacco, in particular diterpenes such as the labdanes and duvanes. It has been assumed that most of the components present in the cuticular surface washes and trichome exudates are synthesized by the trichomes. However, there is little definitive evidence to support this assumption. Utilizing radiolabeled precursors, studies were undertaken to determine the site or sites of 1S- and 1R-4.8, 13-duvatriene-1,3-diol (1S- and 1R-diol) biosynthesis. Experiments using midvein sections of Tobacco Introduction 1068 treated with (2-/sup 14/C)acetate or mevalonic acid indicated that radioactivity was incorporated into surface components, including 1S- and 1R-diol. Subsequent experiments demonstrated that all of the labeled duvatrienediols found were associated with the exudate and surface extracts. Experiments using incubated detached glandular trichome heads unequivocally demonstrated that the glandular heads have the biosynthetic capacity to incorporate (2-/sup 14/C)acetate or mevalonic acid into 1S- and 1R-diol. The influence of nitrogen fertilization, water stress, time of topping and curing conditions on the accumulated levels of 1S- and 1R-diol in field grown Ky 14 was also examined.

  16. Three new labdanes isolated from Eragrostis viscosa

    Energy Technology Data Exchange (ETDEWEB)

    Sebastiao, N' Soki N. [Chemistry Department, Agostinho Neto University, Luanda (Angola); Fernandes, Nelson; Vieira, Liliana; Mendonca, Dina I.M.D. de [Textile and Paper Materials Center and Chemistry Department, University of Beira Interior, Covilha (Portugal); Mendonca, Antonio J.G. [CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Covilha, (Portugal); Gaspar, Jorge F.; Martins, Celia; Rueff, Jose, E-mail: disabel@ubi.pt [Genetics Department, Faculty of Medical Sciences, New University of Lisbon, Lisboa (Portugal); Diakanamwa, Carlos [Biology Department, Agostinho Neto University, Luanda (Angola)

    2012-10-15

    Three new labdanes with 8{alpha},15-epoxy ring [methyl 8{alpha},15-epoxylabdan-16{beta}-oate, 8{alpha},15-epoxylabdan-16{beta}-ol and 8{alpha},15-epoxy-16-norlabdan-13{beta}-ol] and five known compounds [8{alpha},15-epoxy-16-norlabdan-13-one, 8{alpha},15-epoxylabdan-16{beta}-oic acid, 3{beta}-(3''4''dihydroxy)-(E)-cinnamoyloxylup-20(29)-ene, 3-(2',3',4',6'-tetra-O-acetyl-{beta}-D-glucopyranosyloxy)-{beta}-sitosterol and 16-acetoxy-8{alpha},15-epoxylabdane] were isolated from toluene and dichloromethane extracts of aerial parts of Eragrostis viscosa. The structures of all the compounds were established based on their spectroscopic data and X-ray diffraction analysis of 8{alpha},15-epoxylabdan-16{beta}-ol. It was also studied the genotoxicity of E. viscosa, particularly compounds 16-acetoxy-8{alpha},15-epoxylabdane, 8{alpha},15-epoxy-16-norlabdan-13-one and 8{alpha},15-epoxilabdan-16{beta}-ol, using a cytokinesis-block micronucleus assay and the Ames test to assess mutagenicity. Both assays were negative. Cytotoxicity was also analyzed using an MTT assay, and 8{alpha},15-epoxy-16{beta}-ol was shown to be the most cytotoxic of the compounds tested. E. viscosa extracts were also tested to determine their antioxidant capacities, peroxide values and total phenolic contents. (author)

  17. Diterpene synthases of the biosynthetic system of medicinally active diterpenoids in Marrubium vulgare

    DEFF Research Database (Denmark)

    Zerbe, Philipp; Chiang, Angela; Dullat, Harpreet

    2014-01-01

    Marrubium vulgare (Lamiaceae) is a medicinal plant whose major bioactive compounds, marrubiin and other labdane-related furanoid diterpenoids, have potential applications as anti-diabetics, analgesics or vasorelaxants. Metabolite and transcriptome profiling of M. vulgare leaves identified five...... (+)-copalyl diphosphate synthase, and the functional diTPS pair MvCPS1 and MvELS. In a sequential reaction, MvCPS1 and MvELS produce a unique oxygenated diterpene scaffold 9,13-epoxy-labd-14-ene en route to marrubiin and an array of related compounds. In contrast with previously known diTPSs that introduce...

  18. Functional characterization of wheat copalyl diphosphate synthases sheds light on the early evolution of labdane-related diterpenoid metabolism in the cereals.

    Science.gov (United States)

    Wu, Yisheng; Zhou, Ke; Toyomasu, Tomonobu; Sugawara, Chizu; Oku, Madoka; Abe, Shiho; Usui, Masami; Mitsuhashi, Wataru; Chono, Makiko; Chandler, Peter M; Peters, Reuben J

    2012-12-01

    Two of the most agriculturally important cereal crop plants are wheat (Triticum aestivum) and rice (Oryza sativa). Rice has been shown to produce a number of diterpenoid natural products as phytoalexins and/or allelochemicals--specifically, labdane-related diterpenoids, whose biosynthesis proceeds via formation of an eponymous labdadienyl/copalyl diphosphate (CPP) intermediate (e.g., the ent-CPP of gibberellin phytohormone biosynthesis). Similar to rice, wheat encodes a number of CPP synthases (CPS), and the three CPS characterized to date (TaCPS1-3) all have been suggested to produce ent-CPP. However, several of the downstream diterpene synthases will only react with CPP intermediate of normal or syn, but not ent, stereochemistry, as described in the accompanying report. Investigation of additional CPS did not resolve this issue, as the only other functional synthase (TaCPS4) also produced ent-CPP. Chiral product characterization of all the TaCPS then established that TaCPS2 uniquely produces normal, rather than ent-, CPP, thus, providing a suitable substrate source for the downstream diterpene synthases. Notably, TaCPS2 is most homologous to the similarly stereochemically differentiated syn-CPP synthase from rice (OsCPS4), while the non-inducible TaCPS3 and TaCPS4 cluster with the rice OsCPS1 required for gibberellin phytohormone biosynthesis, as well as with a barley (Hordeum vulgare) CPS (HvCPS1) that also is characterized here as similarly producing ent-CPP. These results suggest that diversification of labdane-related diterpenoid metabolism beyond the ancestral gibberellins occurred early in cereal evolution, and included the type of stereochemical variation demonstrated here.

  19. Absolute configuration of labdane diterpenoids from Physalis nicandroides.

    Science.gov (United States)

    Maldonado, Emma; Pérez-Castorena, Ana L; Romero, Yunuen; Martínez, Mahinda

    2015-02-27

    A mixture of the new epimeric labdenetriols 1 and 2 was isolated from the aerial parts of Physalis nicandroides. The structures of 1 and 2, including their absolute configurations, were established by analyses of their spectroscopic data, together with the X-ray diffraction analysis of acetonide 3 and chemical correlation with (-)-(13E)-labd-13-ene-8α,15-diol (6), whose absolute configuration was also confirmed by X-ray analysis of its dibromo derivative 7. The epimeric labdenediols 8 and 9, the known labdanes 6 and 11, and the acylsucroses 12 and 13 were also isolated. Labdanes 6 and 11 showed moderate anti-inflammatory activities in the induced ear edema model.

  20. An Efficient Cationic Cyclization Approach for the Construction of Labdane Diterpenoid Decalin Ring Skeleton

    Institute of Scientific and Technical Information of China (English)

    Jin Hui YANG; Wei Dong Z.LI

    2005-01-01

    An effective approach for the construction of the decalin ring skeleton of labdane diterpenoids was developed based on a key biomimetic cationic polyene cyclization of an epoxy allylsilane precursor. The synthetic approach demonstrated here would be useful in the enantioselective and diastereoselective total synthesis of natural labdane diterpenoids in general.

  1. Analgesic potential of marrubiin derivatives, a bioactive diterpene present in Marrubium vulgare (Lamiaceae).

    Science.gov (United States)

    Meyre-Silva, C; Yunes, R A; Schlemper, V; Campos-Buzzi, F; Cechinel-Filho, V

    2005-04-01

    Marrubiin, a furane labdane diterpene, is the main analgesic compound present in Marrubium vulgare, a medicinal plant used in Brazil and other countries to treat several ailments. Considering its important pharmacological action, as well as its high yield, some structural modifications were performed in order to obtain more active compounds. Success was obtained in reducing the lactonic function, in the formation of marrubiinic acid and two esterified derivatives, which exhibited significant analgesic effect against the writhing test in mice. Marrubiinic acid showed better activity and excellent yield, and its analgesic effect was confirmed in other experimental models of pain in mice, suggesting its possible use as a model to obtain new and potent analgesic agents.

  2. Extracellular localization of the diterpene sclareol in clary sage (Salvia sclarea L., Lamiaceae.

    Directory of Open Access Journals (Sweden)

    Jean-Claude Caissard

    Full Text Available Sclareol is a high-value natural product obtained by solid/liquid extraction of clary sage (Salvia sclarea L. inflorescences. Because processes of excretion and accumulation of this labdane diterpene are unknown, the aim of this work was to gain knowledge on its sites of accumulation in planta. Samples were collected in natura or during different steps of the industrial process of extraction (steam distillation and solid/liquid extraction. Samples were then analysed with a combination of complementary analytical techniques (gas chromatography coupled to a mass spectrometer, polarized light microscopy, environmental scanning electron microscopy, two-photon fluorescence microscopy, second harmonic generation microscopy. According to the literature, it is hypothesized that sclareol is localized in oil pockets of secretory trichomes. This study demonstrates that this is not the case and that sclareol accumulates in a crystalline epicuticular form, mostly on calyces.

  3. Extracellular localization of the diterpene sclareol in clary sage (Salvia sclarea L., Lamiaceae).

    Science.gov (United States)

    Caissard, Jean-Claude; Olivier, Thomas; Delbecque, Claire; Palle, Sabine; Garry, Pierre-Philippe; Audran, Arthur; Valot, Nadine; Moja, Sandrine; Nicolé, Florence; Magnard, Jean-Louis; Legrand, Sylvain; Baudino, Sylvie; Jullien, Frédéric

    2012-01-01

    Sclareol is a high-value natural product obtained by solid/liquid extraction of clary sage (Salvia sclarea L.) inflorescences. Because processes of excretion and accumulation of this labdane diterpene are unknown, the aim of this work was to gain knowledge on its sites of accumulation in planta. Samples were collected in natura or during different steps of the industrial process of extraction (steam distillation and solid/liquid extraction). Samples were then analysed with a combination of complementary analytical techniques (gas chromatography coupled to a mass spectrometer, polarized light microscopy, environmental scanning electron microscopy, two-photon fluorescence microscopy, second harmonic generation microscopy). According to the literature, it is hypothesized that sclareol is localized in oil pockets of secretory trichomes. This study demonstrates that this is not the case and that sclareol accumulates in a crystalline epicuticular form, mostly on calyces.

  4. Extracellular Localization of the Diterpene Sclareol in Clary Sage (Salvia sclarea L., Lamiaceae)

    Science.gov (United States)

    Caissard, Jean-Claude; Olivier, Thomas; Delbecque, Claire; Palle, Sabine; Garry, Pierre-Philippe; Audran, Arthur; Valot, Nadine; Moja, Sandrine; Nicolé, Florence; Magnard, Jean-Louis; Legrand, Sylvain; Baudino, Sylvie; Jullien, Frédéric

    2012-01-01

    Sclareol is a high-value natural product obtained by solid/liquid extraction of clary sage (Salvia sclarea L.) inflorescences. Because processes of excretion and accumulation of this labdane diterpene are unknown, the aim of this work was to gain knowledge on its sites of accumulation in planta. Samples were collected in natura or during different steps of the industrial process of extraction (steam distillation and solid/liquid extraction). Samples were then analysed with a combination of complementary analytical techniques (gas chromatography coupled to a mass spectrometer, polarized light microscopy, environmental scanning electron microscopy, two-photon fluorescence microscopy, second harmonic generation microscopy). According to the literature, it is hypothesized that sclareol is localized in oil pockets of secretory trichomes. This study demonstrates that this is not the case and that sclareol accumulates in a crystalline epicuticular form, mostly on calyces. PMID:23133579

  5. Diterpene synthases of the biosynthetic system of medicinally active diterpenoids in Marrubium vulgare.

    Science.gov (United States)

    Zerbe, Philipp; Chiang, Angela; Dullat, Harpreet; O'Neil-Johnson, Mark; Starks, Courtney; Hamberger, Björn; Bohlmann, Jörg

    2014-09-01

    Marrubium vulgare (Lamiaceae) is a medicinal plant whose major bioactive compounds, marrubiin and other labdane-related furanoid diterpenoids, have potential applications as anti-diabetics, analgesics or vasorelaxants. Metabolite and transcriptome profiling of M. vulgare leaves identified five different candidate diterpene synthases (diTPSs) of the TPS-c and TPS-e/f clades. We describe the in vitro and in vivo functional characterization of the M. vulgare diTPS family. In addition to MvEKS ent-kaurene synthase of general metabolism, we identified three diTPSs of specialized metabolism: MvCPS3 (+)-copalyl diphosphate synthase, and the functional diTPS pair MvCPS1 and MvELS. In a sequential reaction, MvCPS1 and MvELS produce a unique oxygenated diterpene scaffold 9,13-epoxy-labd-14-ene en route to marrubiin and an array of related compounds. In contrast with previously known diTPSs that introduce a hydroxyl group at carbon C-8 of the labdane backbone, the MvCPS1-catalyzed reaction proceeds via oxygenation of an intermediate carbocation at C-9, yielding the bicyclic peregrinol diphosphate. MvELS belongs to a subgroup of the diTPS TPS-e/f clade with unusual βα-domain architecture. MvELS is active in vitro and in vivo with three different prenyl diphosphate substrates forming the marrubiin precursor 9,13-epoxy-labd-14-ene, as identified by nuclear magnetic resonance (NMR) analysis, manoyl oxide and miltiradiene. MvELS fills a central position in the biosynthetic system that forms the foundation for the diverse repertoire of Marrubium diterpenoids. Co-expression of MvCPS1 and MvELS in engineered E. coli and Nicotiana benthamiana offers opportunities for producing precursors for an array of biologically active diterpenoids.

  6. Diterpene production in Mycobacterium tuberculosis

    Science.gov (United States)

    Prach, Lisa; Kirby, James; Keasling, Jay D.; Alber, Tom

    2011-01-01

    Diterpenes are a structurally diverse class of molecules common in plants, although they are very rarely found in bacteria. We report the identification in Mycobacterium tuberculosis (Mtb) of three diterpenes proposed to promote phagolysosome maturation arrest. MS analysis reveals that these diterpenes are novel compounds not previously identified in other organisms. The diterpene with highest abundance in Mtb has a mass fragmentation pattern identical to edaxadiene, which is produced in vitro from geranylgeranyl diphosphate by the enzymes Rv3377c and Rv3378c [Mann FM et al. (2009) J Am Chem Soc 131, 17526–17527]. A second diterpene found in Mtb has a similar mass spectrum, and is always observed in the same proportion relative to edaxadiene, indicating that it is a side product of the Rv3378c reaction in vivo. We name this second diterpene olefin edaxadiene B. The least abundant of the three diterpenes in Mtb extracts is tuberculosinol, a dephosphorylated side-product of the edaxadiene pathway intermediate produced by Rv3377c [Nakano C et al. (2009) Chembiochem 10, 2060–2071; Nakano C et al. (2005) Chem Commun (Camb) 8, 1016–1018]. A frameshift in Rv3377c in Mtb completely eliminates diterpene production, whereas expression of Rv3377c and Rv3378c in the nonpathogenic M. smegmatis is sufficient to produce edaxadiene and edaxadiene B. These studies define the pathway of edaxadiene and edaxadiene B biosynthesis in vivo. Rv3377c and Rv3378c are unique to Mtb and M. bovis, making them candidates for selective therapeutics and diagnostics. PMID:20670276

  7. Two residues determine the product profile of the class II diterpene synthases TPS14 and TPS21 of Tripterygium wilfordii

    DEFF Research Database (Denmark)

    Hansen, Nikolaj Lervad; Nissen, Jakob N.; Hamberger, Björn Robert

    2017-01-01

    The medicinal plant Tripterygium wilfordii (Celastraceae) contains a pair of class II diterpene synthases (diTPS) of specialized labdane-type metabolism that, despite remarkably close homology, form strikingly different products. TwTPS21 catalyzes bicyclization of the linear C20 precursor...... residue gave mixed product profiles. Two mutants, TwTPS14:Y265H and TwTPS21:A325V, also produced ent-copalyl diphosphate, highlighting the evolutionary potential of enzymes of this family to drive rapid diversification of plant diterpene biosynthesis through neo-functionalization. Our study contributes...... to the understanding of structure-function relation in plant class II diTPSs and complements previous mutational studies of Arabidopsis ent-copalyl diphosphate synthase with additional examples from the specialized metabolism of T. wilfordii....

  8. Discovery and functional characterization of two diterpene synthases for sclareol biosynthesis in Salvia sclarea (L.) and their relevance for perfume manufacture.

    Science.gov (United States)

    Caniard, Anne; Zerbe, Philipp; Legrand, Sylvain; Cohade, Allison; Valot, Nadine; Magnard, Jean-Louis; Bohlmann, Jörg; Legendre, Laurent

    2012-07-26

    Sclareol is a diterpene natural product of high value for the fragrance industry. Its labdane carbon skeleton and its two hydroxyl groups also make it a valued starting material for semisynthesis of numerous commercial substances, including production of Ambrox® and related ambergris substitutes used in the formulation of high end perfumes. Most of the commercially-produced sclareol is derived from cultivated clary sage (Salvia sclarea) and extraction of the plant material. In clary sage, sclareol mainly accumulates in essential oil-producing trichomes that densely cover flower calices. Manool also is a minor diterpene of this species and the main diterpene of related Salvia species. Based on previous general knowledge of diterpene biosynthesis in angiosperms, and based on mining of our recently published transcriptome database obtained by deep 454-sequencing of cDNA from clary sage calices, we cloned and functionally characterized two new diterpene synthase (diTPS) enzymes for the complete biosynthesis of sclareol in clary sage. A class II diTPS (SsLPPS) produced labda-13-en-8-ol diphosphate as major product from geranylgeranyl diphosphate (GGPP) with some minor quantities of its non-hydroxylated analogue, (9 S, 10 S)-copalyl diphosphate. A class I diTPS (SsSS) then transformed these intermediates into sclareol and manool, respectively. The production of sclareol was reconstructed in vitro by combining the two recombinant diTPS enzymes with the GGPP starting substrate and in vivo by co-expression of the two proteins in yeast (Saccharomyces cerevisiae). Tobacco-based transient expression assays of green fluorescent protein-fusion constructs revealed that both enzymes possess an N-terminal signal sequence that actively targets SsLPPS and SsSS to the chloroplast, a major site of GGPP and diterpene production in plants. SsLPPS and SsSS are two monofunctional diTPSs which, together, produce the diterpenoid specialized metabolite sclareol in a two-step process. They

  9. Discovery and functional characterization of two diterpene synthases for sclareol biosynthesis in Salvia sclarea (L. and their relevance for perfume manufacture

    Directory of Open Access Journals (Sweden)

    Caniard Anne

    2012-07-01

    Full Text Available Abstract Background Sclareol is a diterpene natural product of high value for the fragrance industry. Its labdane carbon skeleton and its two hydroxyl groups also make it a valued starting material for semisynthesis of numerous commercial substances, including production of Ambrox® and related ambergris substitutes used in the formulation of high end perfumes. Most of the commercially-produced sclareol is derived from cultivated clary sage (Salvia sclarea and extraction of the plant material. In clary sage, sclareol mainly accumulates in essential oil-producing trichomes that densely cover flower calices. Manool also is a minor diterpene of this species and the main diterpene of related Salvia species. Results Based on previous general knowledge of diterpene biosynthesis in angiosperms, and based on mining of our recently published transcriptome database obtained by deep 454-sequencing of cDNA from clary sage calices, we cloned and functionally characterized two new diterpene synthase (diTPS enzymes for the complete biosynthesis of sclareol in clary sage. A class II diTPS (SsLPPS produced labda-13-en-8-ol diphosphate as major product from geranylgeranyl diphosphate (GGPP with some minor quantities of its non-hydroxylated analogue, (9 S, 10 S-copalyl diphosphate. A class I diTPS (SsSS then transformed these intermediates into sclareol and manool, respectively. The production of sclareol was reconstructed in vitro by combining the two recombinant diTPS enzymes with the GGPP starting substrate and in vivo by co-expression of the two proteins in yeast (Saccharomyces cerevisiae. Tobacco-based transient expression assays of green fluorescent protein-fusion constructs revealed that both enzymes possess an N-terminal signal sequence that actively targets SsLPPS and SsSS to the chloroplast, a major site of GGPP and diterpene production in plants. Conclusions SsLPPS and SsSS are two monofunctional diTPSs which, together, produce the diterpenoid

  10. Discovery and functional characterization of two diterpene synthases for sclareol biosynthesis in Salvia sclarea (L.) and their relevance for perfume manufacture

    Science.gov (United States)

    2012-01-01

    Background Sclareol is a diterpene natural product of high value for the fragrance industry. Its labdane carbon skeleton and its two hydroxyl groups also make it a valued starting material for semisynthesis of numerous commercial substances, including production of Ambrox® and related ambergris substitutes used in the formulation of high end perfumes. Most of the commercially-produced sclareol is derived from cultivated clary sage (Salvia sclarea) and extraction of the plant material. In clary sage, sclareol mainly accumulates in essential oil-producing trichomes that densely cover flower calices. Manool also is a minor diterpene of this species and the main diterpene of related Salvia species. Results Based on previous general knowledge of diterpene biosynthesis in angiosperms, and based on mining of our recently published transcriptome database obtained by deep 454-sequencing of cDNA from clary sage calices, we cloned and functionally characterized two new diterpene synthase (diTPS) enzymes for the complete biosynthesis of sclareol in clary sage. A class II diTPS (SsLPPS) produced labda-13-en-8-ol diphosphate as major product from geranylgeranyl diphosphate (GGPP) with some minor quantities of its non-hydroxylated analogue, (9 S, 10 S)-copalyl diphosphate. A class I diTPS (SsSS) then transformed these intermediates into sclareol and manool, respectively. The production of sclareol was reconstructed in vitro by combining the two recombinant diTPS enzymes with the GGPP starting substrate and in vivo by co-expression of the two proteins in yeast (Saccharomyces cerevisiae). Tobacco-based transient expression assays of green fluorescent protein-fusion constructs revealed that both enzymes possess an N-terminal signal sequence that actively targets SsLPPS and SsSS to the chloroplast, a major site of GGPP and diterpene production in plants. Conclusions SsLPPS and SsSS are two monofunctional diTPSs which, together, produce the diterpenoid specialized metabolite sclareol

  11. Suppression of inflammatory responses by labdane-type diterpenoids.

    Science.gov (United States)

    Girón, Natalia; Través, Paqui G; Rodríguez, Benjamín; López-Fontal, Raquel; Boscá, Lisardo; Hortelano, Sonsoles; de las Heras, Beatriz

    2008-04-15

    A series of 11 labdane-type diterpenoids (1-11) with various patterns of substitution were tested for potential anti-inflammatory activity. Of these compounds, 4 and 11 were selected to evaluate their influence on targets relevant to the regulation of the inflammatory response. These diterpenoids reduced the production of nitric oxide (NO), prostaglandin E2, and tumor necrosis factor-alpha in LPS-activated RAW 264.7 macrophages, with IC50 in the range 1-10 microM. Inhibition of these inflammatory mediators was related to inhibition of the expression of nitric oxide synthase-2 (NOS-2) and cyclooxygenase-2 (COX-2) at the transcriptional level, as determined by western-blot and RT-PCR. Examination of the effects of these diterpenoids on nuclear factor kappaB signaling showed that both compounds inhibit the phosphorylation of IkappaBalpha and IkappaBbeta, preventing their degradation and the nuclear translocation of the NF-kappaB p65 subunit. Inhibition of IKK activity was also observed. These derivatives displayed significant anti-inflammatory activity in vivo, suppressing mouse ear edema induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) and inhibiting myeloperoxidase activity, an index of neutrophil infiltration. The anti-inflammatory effects of these labdane diterpenoids, together with their low cell toxicity, suggest potential therapeutic applications in the regulation of the inflammatory response.

  12. Evaluation of labdane derivatives as potential anti-inflammatory agents.

    Science.gov (United States)

    Girón, Natalia; Pérez-Sacau, Elisa; López-Fontal, Raquel; Amaro-Luis, Juan M; Hortelano, Sonsoles; Estevez-Braun, Ana; de Las Heras, Beatriz

    2010-07-01

    In the present study, a series of labdane derivatives (2-9) were prepared from labdanediol (1) and their potential as anti-inflammatory agents were evaluated on lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. All compounds were able to inhibit LPS-induced nitric oxide (NO), although compounds 1, 2, 5, 8 and 9 exhibited the most potent effects with a range of IC(50) values of 5-15 microM. Similarly to the inhibitory effects on NO release, these labdane derivatives also inhibited prostaglandin E(2) (PGE(2)) production. However, analysis of cell viability demonstrated that effects on NO release and (PGE(2)) production of compounds 1, 8 and 9 were due to citotoxicity, whereas compound 2 and 5 did not show any effect in the survival of RAW 264.7 macrophages. In addition to these in vitro data, compound 5 also showed anti-inflammatory activity in vivo, when tested in mice. They prevented the extent of swelling in the TPA-induced ear edema model and inhibited MPO activity, showing similar potency to that of the widely used anti-inflammatory drug indomethacin. These results indicate that compound 2 and in particular compound 5 might be used for the design of new anti-inflammatory agents. Copyright (c) 2010 Elsevier Masson SAS. All rights reserved.

  13. Oxidative Degradations of the Side Chain of Unsaturated Ent-labdanes. Part II

    Directory of Open Access Journals (Sweden)

    María Cristina Chamy

    2007-12-01

    Full Text Available A route for the degradation of the side chain of ent-labdane derivatives has beendevised, giving the useful synthon 2β,12-dihydroxy-13,14,15,16,17-pentanor-ent-labdane-8-one (8. The use of this compound in the preparation of terpenylquinone derivatives shallbe reported elsewhere. In addition we have synthesized the compound 2β,12-diacetoxy-8β,17-epoxy-13,14,15,16-tetranor-ent-labdane (10, which upon catalytic epoxide ringopening in alkaline or acid media gave rise in all cases to the formation of tricycliccompounds.

  14. Increasing diterpene yield with a modular metabolic engineering system in E. coli: comparison of MEV and MEP isoprenoid precursor pathway engineering.

    Science.gov (United States)

    Morrone, Dana; Lowry, Luke; Determan, Mara K; Hershey, David M; Xu, Meimei; Peters, Reuben J

    2010-02-01

    Engineering biosynthetic pathways in heterologous microbial host organisms offers an elegant approach to pathway elucidation via the incorporation of putative biosynthetic enzymes and characterization of resulting novel metabolites. Our previous work in Escherichia coli demonstrated the feasibility of a facile modular approach to engineering the production of labdane-related diterpene (20 carbon) natural products. However, yield was limited (engineering system. With MEP pathway enhancement, it was found that pyruvate supplementation of rich media and simultaneous overexpression of three genes (idi, dxs, and dxr) resulted in the greatest increase in diterpene yield, indicating distributed metabolic control within this pathway. Incorporation of a heterologous MEV pathway in bioreactor grown cultures resulted in significantly higher yields than MEP pathway enhancement. We have established suitable growth conditions for diterpene production levels ranging from 10 to >100 mg/L of E. coli culture. These amounts are sufficient for nuclear magnetic resonance analyses, enabling characterization of enzymatic products and hence, pathway elucidation. Furthermore, these results represent an up to >1,000-fold improvement in diterpene production from our facile, modular platform, with MEP pathway enhancement offering a cost effective alternative with reasonable yield. Finally, we reiterate here that this modular approach is expandable and should be easily adaptable to the production of any terpenoid natural product.

  15. Novel diterpene from Dollabella californica

    Energy Technology Data Exchange (ETDEWEB)

    Ireland, C. (Scripps Institution of Oceanography, La Jolla, CA); Faulkner, D.J.; Finer, J.; Clardy, J.

    1976-07-21

    The sea hare Dollabella californica Sterns, a large softbodied opistobranch mollusk, was collected at Isla Partida, Gulf of California. The digestive gland of Dollabella, contained a number of terpenoid compounds which are probably of dietary origin. The major components of the digestive gland extracts are a series of diterpenes which appear to be closely related. The structural determination has been obtained by single-crystal x-ray diffraction analysis for a diterpene 1 having a novel 5,11-bicyclic carbon skeleton. (DDA)

  16. Oxidative Degradations of the Side Chain of Unsaturated Ent-labdanes. Part I.

    Directory of Open Access Journals (Sweden)

    Karen Catalán Marín

    2007-03-01

    Full Text Available A selective route for the degradation of the unsaturated side chain of ent-labdanes has been devised, giving two useful synthons: 2β-acetoxy-14,15,17-trinor-ent-labdane-8,13- dione (5 and 2β-acetoxy-14,15-dinor-ent-labd-8(17-en-13-one (7, the use of which for the preparation of terpenylquinone derivatives shall be reported elsewhere.

  17. Biochemical characterization of the castor bean ent-kaurene synthase(-like) family supports quantum chemical view of diterpene cyclization.

    Science.gov (United States)

    Jackson, Alana J; Hershey, David M; Chesnut, Taylor; Xu, Meimei; Peters, Reuben J

    2014-07-01

    It has become apparent that plants have extensively diversified their arsenal of labdane-related diterpenoids (LRDs), in part via gene duplication and neo-functionalization of the ancestral ent-kaurene synthase (KS) required for gibberellin metabolism. For example, castor bean (Ricinus communis) was previously shown to produce an interesting set of biosynthetically related diterpenes, specifically ent-sandracopimaradiene, ent-beyerene, and ent-trachylobane, in addition to ent-kaurene, using four separate diterpene synthases, albeit these remain unidentified. Notably, despite mechanistic similarity of the underlying reaction to that catalyzed by KSs, ent-beyerene and ent-trachylobane synthases have not yet been identified. Given our interest in LRD biosynthesis, and the recent availability of the castor bean genome sequence, a synthetic biology approach was applied to biochemically characterize the four KS(-like) enzymes [KS(L)s] found in Ricinus communis [i.e., the RcKS(L)s]. In particular, using bacteria engineered to produce the relevant ent-copalyl diphosphate precursor and synthetic genes based on the predicted RcKS(L)s, although this ultimately required correction of a "splicing" error in one of the predicted genes, highlighting the dependence of such a synthetic biology approach on accurate gene sequences. Nevertheless, it is possible to assign each of the four RcKS(L)s to one of the previously observed diterpene synthase activities, providing access to functionally enzymes. Intriguingly, the product distribution of the RcKS(L)s seems to support the distinct diterpene synthase reaction mechanism proposed by quantum chemical calculations, rather than the classically proposed pathway.

  18. Biochemical characterization of the castor bean ent-kaurene synthase(-like) family supports quantum chemical view of diterpene cyclization

    Science.gov (United States)

    Jackson, Alana J.; Hershey, David M.; Chesnut, Taylor; Xu, Meimei; Peters, Reuben J.

    2014-01-01

    It has become apparent that plants have extensively diversified their arsenal of labdane-related diterpenoids (LRDs), in part via gene duplication and neo-functionalization of the ancestral ent-kaurene synthase (KS) required for gibberellin metabolism. For example, castor bean (Ricinus communis) was previously shown to produce an interesting set of biosynthetically related diterpenes, specifically ent-sandracopimaradiene, ent-beyerene, and ent-trachylobane, in addition to ent-kaurene, using four separate diterpene synthases, albeit these remain unidentified. Notably, despite mechanistic similarity of the underlying reaction to that catalyzed by KSs, ent-beyerene and ent-trachylobane synthases have not yet been identified. Given our interest in LRD biosynthesis, and the recent availability of the castor bean genome sequence, we applied a synthetic biology approach to biochemically characterize the four KS(-like) enzymes [KS(L)s] found in Ricinus communis [i.e., the RcKS(L)s]. In particular, using bacteria engineered to produce the relevant ent-copalyl diphosphate precursor and synthetic genes based on the predicted RcKS(L)s, although this ultimately required correction of a “splicing” error in one of the predicted genes, highlighting the dependence of such a synthetic biology approach on accurate gene sequences. Nevertheless, we can assign each of the four RcKS(L)s to one of the previously observed diterpene synthase activities, providing access to functionally novel enzymes. Intriguingly, the product distribution of the RcKS(L)s seems to support the distinct diterpene synthase reaction mechanism proposed by quantum chemical calculations, rather than the classically proposed pathway. PMID:24810014

  19. Cell growth inhibitory action of an unusual labdane diterpene, 13-epi-sclareol in breast and uterine cancers in vitro.

    Science.gov (United States)

    Sashidhara, Koneni V; Rosaiah, Jammikuntla N; Kumar, Abdhesh; Bid, Hemant K; Konwar, Rituraj; Chattopadhyay, Naibedya

    2007-11-01

    In the course of our studies on the isolation of bioactive compounds from the roots of Coleus forskohlii, a traditional herb in India, rare 13-epi-sclareol has been isolated, and its structure determined by extensive 2D NMR. This is the first report of isolation from this plant. The isolated compound showed antiproliferative activity in breast and uterine cancers in vitro. The antiproliferative activity of 13-epi-sclareol is comparable to Tamoxifen in terms of IC50 and also showed concentration dependent increased apoptotic changes in the breast cancer cell line, MCF-7.

  20. A Single Residue Switch for Mg2+-dependent Inhibition Characterizes Plant Class II Diterpene Cyclases from Primary and Secondary Metabolism*

    Science.gov (United States)

    Mann, Francis M.; Prisic, Sladjana; Davenport, Emily K.; Determan, Mara K.; Coates, Robert M.; Peters, Reuben J.

    2010-01-01

    Class II diterpene cyclases mediate the acid-initiated cycloisomerization reaction that serves as the committed step in biosynthesis of the large class of labdane-related diterpenoid natural products, which includes the important gibberellin plant hormones. Intriguingly, these enzymes are differentially susceptible to inhibition by their Mg2+ cofactor, with those involved in gibberellin biosynthesis being more sensitive to such inhibition than those devoted to secondary metabolism, which presumably limits flux toward the potent gibberellin phytohormones. Such inhibition has been suggested to arise from intrasteric Mg2+ binding to the DXDD motif that cooperatively acts as the catalytic acid, whose affinity must then be modulated in some fashion. While further investigating class II diterpene cyclase catalysis, we discovered a conserved basic residue that seems to act as a counter ion to the DXDD motif, enhancing the ability of aspartic acid to carry out the requisite energetically difficult protonation of a carbon-carbon double bond and also affecting inhibitory Mg2+ binding. Notably, this residue is conserved as a histidine in enzymes involved in gibberellin biosynthesis and as an arginine in those dedicated to secondary metabolism. Interchanging the identity of these residues is sufficient to switch the sensitivity of the parent enzyme to inhibition by Mg2+. These striking findings indicate that this is a single residue switch for Mg2+ inhibition, which not only supports the importance of this biochemical regulatory mechanism in limiting gibberellin biosynthesis, but the importance of its release, presumably to enable higher flux, into secondary metabolism. PMID:20430888

  1. Abietane diterpenes from Hyptis platanifolia

    Energy Technology Data Exchange (ETDEWEB)

    Araujo, Edigenia C.C.; Lima, Mary Anne S.; Silveira, Edilberto R. [Ceara Univ., Fortaleza, CE (Brazil). Dept. de Quimica Organica e Inorganica. Curso de Pos-Graduacao em Quimica Organica]. E-mail: edil@ufc.br; Nunes, Edson P. [Ceara Univ., Fortaleza, CE (Brazil). Dept. de Biologia

    2005-11-15

    Three new abietane diterpenes 19-oxo-inuroyleanol, 11,14-dihydroxy-12-methoxy-7-oxo-8,11,13-abietatriene-19,20 {beta}-olide and 19,20-epoxy-12-methoxy-11,14,19-trihydroxy-7-oxo-8,11,13 -abietatriene, in addition to inuroyleanol, and coulterone were isolated from roots of Hyptis platanifolia. Investigation of the aerial parts yielded a mixture of the steroids stigmasterol and {beta}-sitosterol, and the triterpenes betulinic and ursolic acids. Structural characteriza of all compounds was established on the basis of spectroscopic methods, particularly 1D and 2D NMR, and comparison with data from literature. (author)

  2. Seasonal Variation of Cistus ladanifer L. Diterpenes

    Science.gov (United States)

    Alías, Juan Carlos; Sosa, Teresa; Valares, Cristina; Escudero, José Carlos; Chaves, Natividad

    2012-01-01

    The exudate of Cistus ladanifer L. consists mainly of two families of secondary metabolites: flavonoids and diterpenes. The amount of flavonoids present in the leaves has a marked seasonal variation, being maximum in summer and minimum in winter. In the present study, we demonstrate that the amount of diterpenes varies seasonally, but with a different pattern: maximum concentration in winter and minimum in spring-summer. The experiments under controlled conditions have shown that temperature influences diterpene production, and in particular, low temperatures. Given this pattern, the functions that these compounds perform in C. ladanifer are probably different. PMID:27137636

  3. Seasonal Variation of Cistus ladanifer L. Diterpenes

    Directory of Open Access Journals (Sweden)

    Juan Carlos Alías

    2012-07-01

    Full Text Available The exudate of Cistus ladanifer L. consists mainly of two families of secondary metabolites: flavonoids and diterpenes. The amount of flavonoids present in the leaves has a marked seasonal variation, being maximum in summer and minimum in winter. In the present study, we demonstrate that the amount of diterpenes varies seasonally, but with a different pattern: maximum concentration in winter and minimum in spring-summer. The experiments under controlled conditions have shown that temperature influences diterpene production, and in particular, low temperatures. Given this pattern, the functions that these compounds perform in C. ladanifer are probably different.

  4. Involvement of an ent-copalyl diphosphate synthase in tissue-specific accumulation of specialized diterpenes in Andrographis paniculata.

    Science.gov (United States)

    Misra, Rajesh Chandra; Garg, Anchal; Roy, Sudeep; Chanotiya, Chandan Singh; Vasudev, Prema G; Ghosh, Sumit

    2015-11-01

    Ent-labdane-related diterpene (ent-LRD) specialized (i.e. secondary) metabolites of the medicinal plant kalmegh (Andrographis paniculata) have long been known for several pharmacological activities. However, our understanding of the ent-LRD biosynthetic pathway has remained largely incomplete. Since ent-LRDs accumulate in leaves, we carried out a comparative transcriptional analysis using leaf and root tissues, and identified 389 differentially expressed transcripts, including 223 transcripts that were preferentially expressed in leaf tissue. Analysis of the transcripts revealed various specialized metabolic pathways, including transcripts of the ent-LRD biosynthetic pathway. Two class II diterpene synthases (ApCPS1 and ApCPS2) along with one (ApCPS1') and two (ApCPS2' and ApCPS2″) transcriptional variants that were the outcomes of alternative splicing of the precursor mRNA and alternative transcriptional termination, respectively, were identified. ApCPS1 and ApCPS2 encode for 832- and 817-amino acids proteins, respectively, and are phylogenetically related to the dicotyledons ent-copalyl diphosphate synthases (ent-CPSs). The spatio-temporal patterns of ent-LRD metabolites accumulation and gene expression suggested a likely role for ApCPS1 in general (i.e. primary) metabolism, perhaps by providing precursor for the biosynthesis of phytohormone gibberellin (GA). However, ApCPS2 is potentially involved in tissue-specific accumulation of ent-LRD specialized metabolites. Bacterially expressed recombinant ApCPS2 catalyzed the conversion of (E,E,E)-geranylgeranyl diphosphate (GGPP), the general precursor of diterpenes to ent-copalyl diphosphate (ent-CPP), the precursor of ent-LRDs. Taken together, these results advance our understanding of the tissue-specific accumulation of specialized ent-LRDs of medicinal importance.

  5. A new diterpene glycoside from Stevia rebaudiana

    National Research Council Canada - National Science Library

    Chaturvedula, Venkata Sai Prakash; Prakash, Indra

    2011-01-01

    From the commercial extract of the leaves of Stevia rebaudiana, a new diterpene glycoside was isolated besides the known steviol glycosides including stevioside, rebaudiosides A-F, rubusoside and dulcoside...

  6. Anxiolytic Activity of Diterpene Alkaloid Songorine.

    Science.gov (United States)

    Nesterova, Yu V; Povet'eva, T N; Suslov, N I; Shults, E E; Ziuz'kov, G N; Aksinenko, S G; Afanas'eva, O G; Krapivin, A V; Kharina, T G

    2015-09-01

    Antianxiety action of diterpene alkaloid songorine was studied using Vogel conflict test. Songorine in a dose of 0.25 mg/kg demonstrated high anxiolytic activity comparable to that of phenazepam and produced no sedative effect.

  7. Two new brominated diterpenes from Laurencia decumbens

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Two new brominated diterpenes, namely, laurendecumtriol and 11-deacetylpinnaterpene C, were isolated and identified from the marine red alga Laurencia decumbens. Their structures were established on the basis of various NMR spectroscopic techniques and HR-ESI-MS analyses.

  8. A New Daphnane Diterpene from Daphne tangutica

    Institute of Scientific and Technical Information of China (English)

    Li PAN; Xiao Feng ZHANG; Hai Feng WU; Li Sheng DING

    2006-01-01

    A new daphnane diterpene was isolated from the root barks of Daphne tangutica Maxim.Its structure was elucidated as 1, 2α-dihydro-20-palimoyldaphnetoxin by the spectroscopic evidence including 2D-NMR.

  9. Diterpenes inhibiting NO production from Euphorbia helioscopia.

    Science.gov (United States)

    Chen, Hongqiang; Wang, Hao; Yang, Bo; Jin, Da-Qing; Yang, Shuliang; Wang, Meicheng; Xu, Jing; Ohizumi, Yasushi; Guo, Yuanqiang

    2014-06-01

    Three new jatrophane diterpenes (1-3), an unreported spectroscopic data jatrophane diterpenene (4), and nine known analogues (5-13) have been isolated from the whole plants of Euphorbia helioscopia. Their structures were established by detailed spectroscopic data analyses (IR, ESIMS, HR-ESIMS, and 1D and 2D NMR), and the structure of 1 was confirmed by X-ray crystallography. The diterpenes showed inhibitory activities on LPS-induced NO production in murine microglial BV-2 cells.

  10. Diterpene Glycosides from Stevia rebaudiana

    Directory of Open Access Journals (Sweden)

    Indra Prakash

    2011-04-01

    Full Text Available Three novel diterpene glycosides were isolated for the first time from the commercial extract of the leaves of Stevia rebaudiana, along with several known steviol glycosides, namely stevioside, rebaudiosides A-F, rubusoside and dulcoside A. The new compounds were identified as 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyloxy] ent-kaur-15-en-19-oic acid (1, 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyloxy]-16β-hydroxy-ent-kauran-19-oic acid (2 and 13-methyl-16-oxo-17-nor-ent-kauran-19-oic acid-β-D-glucopyranosyl ester (3 on the basis of extensive 2D NMR and MS spectroscopic data as well as chemical studies.

  11. Clerodane diterpenes from bark of Croton urucurana baillon

    Energy Technology Data Exchange (ETDEWEB)

    Pizzolatti, Moacir G.; Bortoluzzi, Adailton J.; Brighente, Ines M.C.; Zuchinalli, Analice; Carvalho, Francieli K., E-mail: moacir.pizzolatti@ufsc.br [Universidade Federal de Santa Catarina (UFSC), Florianopolis, SC (Brazil). Departamento de Qumica; Candido, Ana C. S.; Peres, Marize T.L.P. [Universidade Federal do Mato Grosso do Sul (UFMS), Campo Grande, MS (Brazil). Departamento de Hidraulica e Transportes

    2013-04-15

    The new clerodane diterpene methyl 3-oxo-12-epibarbascoate was isolated from the stem barks of Croton urucurana together with the known diterpene methyl 12-epibarbascoate. The structures of these compounds were elucidated by spectroscopic techniques and comparison with the literature data. The obtainment of crystals allowed the crystallographic analysis of X-ray diffraction of diterpenes, thus confirming the proposed structures. (author)

  12. New diterpenes from the heartwood of Chamaecyparis obtusa var. formosana.

    Science.gov (United States)

    Kuo, Y H; Chen, C H; Huang, S L

    1998-06-26

    An abietane diterpene, 11,14-dihydroxy-8,11,13-abietatrien-7-one (1); a seco-abietane diterpene, obtuanhydride (2); and an isopimarane diterpene, 18,19-O-isopropylidene-18, 19-dihydroxyisopimara-8(14),15-diene (3) were isolated from the heartwood of Chamaecyparis obtusa var. formosana. The structures of these new compounds were elucidated by spectroscopic methods.

  13. New labdane diterpenoids from Croton laui and their anti-inflammatory activities.

    Science.gov (United States)

    Yang, Li; Zhang, Yu-Bo; Chen, Li-Feng; Chen, Neng-Hua; Wu, Zhong-Nan; Jiang, Si-Qi; Jiang, Lin; Li, Guo-Qiang; Li, Yao-Lan; Wang, Guo-Cai

    2016-10-01

    Nine new labdane diterpenoids (1-9) were isolated from the aerial parts of Croton laui, along with eight known analogues (10-17). Their structures were identified on the basis of the spectral data (IR, UV, HRESIMS, 1D and 2D NMR), and the structure of 8 was confirmed by single crystal X-ray diffraction analyses. In addition, compounds 1, 4, 7, 8, and 14 showed weak anti-inflammatory activities in LPS-stimulated RAW 264.7 cells.

  14. A new casbane diterpene from Euphorbia pekinensis.

    Science.gov (United States)

    Wang, Kuilong; Yu, Hongli; Wu, Hao; Wang, Xinzhi; Pan, Yaozong; Chen, Yeqing; Liu, Liping; Jin, Yangping; Zhang, Chenchao

    2015-01-01

    A new casbane diterpenoid, referred to as pekinenin G, together with one cembrane diterpene and four known casbane diterpenoids were isolated from the roots of Euphorbia pekinensis. Their structures were elucidated on the basis of spectroscopic studies and comparison with related known compounds. The six compounds showed different cytotoxic activities against four human cancer cell lines.

  15. New Myrsinol Diterpenes from Euphorbia prolifera

    Institute of Scientific and Technical Information of China (English)

    ZHANG,Wan-Jin(张万金); CHEN,Dao-Feng(陈道峰); HOU,Ai-Jun(侯爱君)

    2004-01-01

    Four new myrsinol diterpenes, euphorprolitherins A-D (1-3, 5), were isolated from the ethanolic extract of the root of Euphorbia prolifera, along with a known one, SPr5 (4). The structures were elucidated on the basis of their spectroscopic evidences. The structure and relative stereochemistry of 1 were confirmed by X-ray crystallography.

  16. A New Diterpene Glycoside from Isodon forrestii

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A new diterpene glycoside named 9(10?20)-abeo-7b,10b,11,15-tetrahydroxy-8,11,13- abietatrien-12-O-b-D-glucopyranoside was isolated from aerial parts of Isodon forrestii. Its structure was determined by means of spectroscopic studies.

  17. Miltiorins A-D, diterpenes from Radix Salviae miltiorrhizae.

    Science.gov (United States)

    Hirata, Ai; Kim, Sang-Yong; Kobayakawa, Natsuki; Tanaka, Naonobu; Kashiwada, Yoshiki

    2015-04-01

    Constituents of the anti-influenza A neuraminidase (NA) active extract from Radix Salviae miltiorrhizae were investigated, resulting in the isolation of four new diterpenes, miltiorins A-D (1-4), together with eight known diterpenes. The structures of 1-4 were assigned by spectroscopic analysis. Miltiorins A-C (1-3) were abietane diterpenes possessing a 2α-acetoxy group and a 12-hydroxy group in common, while miltiorin D (4) was a 11,12-seco-abietane diterpene with a γ-lactone ring. Miltiorin D (4) is the first example of a 11,12-seco-abietane diterpene from natural sources. Anti-NA activities of the isolated diterpenes were evaluated.

  18. Gene discovery of modular diterpene metabolism in nonmodel systems.

    Science.gov (United States)

    Zerbe, Philipp; Hamberger, Björn; Yuen, Macaire M S; Chiang, Angela; Sandhu, Harpreet K; Madilao, Lina L; Nguyen, Anh; Hamberger, Britta; Bach, Søren Spanner; Bohlmann, Jörg

    2013-06-01

    Plants produce over 10,000 different diterpenes of specialized (secondary) metabolism, and fewer diterpenes of general (primary) metabolism. Specialized diterpenes may have functions in ecological interactions of plants with other organisms and also benefit humanity as pharmaceuticals, fragrances, resins, and other industrial bioproducts. Examples of high-value diterpenes are taxol and forskolin pharmaceuticals or ambroxide fragrances. Yields and purity of diterpenes obtained from natural sources or by chemical synthesis are often insufficient for large-volume or high-end applications. Improvement of agricultural or biotechnological diterpene production requires knowledge of biosynthetic genes and enzymes. However, specialized diterpene pathways are extremely diverse across the plant kingdom, and most specialized diterpenes are taxonomically restricted to a few plant species, genera, or families. Consequently, there is no single reference system to guide gene discovery and rapid annotation of specialized diterpene pathways. Functional diversification of genes and plasticity of enzyme functions of these pathways further complicate correct annotation. To address this challenge, we used a set of 10 different plant species to develop a general strategy for diterpene gene discovery in nonmodel systems. The approach combines metabolite-guided transcriptome resources, custom diterpene synthase (diTPS) and cytochrome P450 reference gene databases, phylogenies, and, as shown for select diTPSs, single and coupled enzyme assays using microbial and plant expression systems. In the 10 species, we identified 46 new diTPS candidates and over 400 putatively terpenoid-related P450s in a resource of nearly 1 million predicted transcripts of diterpene-accumulating tissues. Phylogenetic patterns of lineage-specific blooms of genes guided functional characterization.

  19. Clerodane diterpenes from leaves of Casearia sylvestris Swartz

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Andre G. dos; Perez, Carla C.; Tininis, Aristeu G.; Bolzani, Vanderlan da S.; Cavalheiro, Alberto J. [UNESP, Araraquara, SP (Brazil). Inst. de Quimica]. E-mail: albjcava@iq.unesp.br

    2007-09-15

    Ethanolic extracts of the leaves of Casearia sylvestris yielded a novel clerodane diterpene, 15-hydroxy-3-cleroden-2-one, together with the known diterpenes (-)-hardwickiic acid, reported for the first time from this species, and casearins B and G, previously isolated from C. sylvestris. The structures of all four compounds were determined by spectrometric analysis. The new clerodane diterpene and (-)-hardwickiic acid contain structural features that are completely different from the highly oxygenated casearins and casearvestrins isolated from C. sylvestris. (author)

  20. Pyrrolizidine alkaloids and diterpenes from Villasenoria orcuttii

    Energy Technology Data Exchange (ETDEWEB)

    Arciniegas, Amira; Perez-Castorena, Ana L.; Gonzalez, Karina; Vivar, Alfonso Romo de, E-mail: alperezc@unam.mx [Instituto de Quimica, Universidad Nacional Autonoma de Mexico, Circuito Exterior, Ciudad Universitaria, Coyoacan, DF (Mexico); Reyes-Lezama, Marisol [Centro Conjunto de Investigacion en Quimica Sustentable, Universidad Autonoma del Estado de Mexico-Universidad Nacional Autonoma de Mexico (UAEM-UNAM), Carretera Toluca-Atlacomulco, Estado de Mexico (Mexico); Villasenor, Jose Luis [Instituto de Biologia, Universidad Nacional Autonoma de Mexico, Circuito Exterior, Ciudad Universitaria, Coyoacan, DF, (Mexico)

    2013-07-15

    The chemical study of Villasenoria orcuttii, the only species of the genus Villasenoria, afforded three acyclic diterpenes, two of them described for the first time. Two pyrrolizidine alkaloids, florosenine and floridanine, among other known compounds were also isolated. The absolute configuration of floridanine was determined by X-ray analysis using anomalous dispersion with Cu K{sub {alpha}} radiation, and its {sup 1}H and {sup 13}C nuclear magnetic resonance (NMR) data were corrected. (author)

  1. Two new diterpene alkaloids from Delphinium chrysotrichum

    Institute of Scientific and Technical Information of China (English)

    Yang Qing He; Xiao Mei Wei; Yi Li Han; Li Ming Gao

    2007-01-01

    Chemical investigation on the ethanol extract from the whole plants of Delphinium chrysotrichum resulted in the isolation of two new diterpene alkaloids named delphatisine A (1) and delphatisine B (2), respectively. The structures of the new compounds were deduced on the basis of their spectral data (IR, HREIMS, EIMS, 1D, 2D-NMR). This is the first report on the isolation of diterpenoid alkaloids from the D. Chrysotrichum.

  2. Two new antitumor diterpenes from Pinus sylvestris

    Institute of Scientific and Technical Information of China (English)

    Qiang Wang; Zhang Rong; Jie Teng; Yoshihisa Takaishi; Hong Quan Duan

    2008-01-01

    Two new diterpenes,15-ethyl- 18-methyl pinifolate (1)and 18-hydroxy-labda-8(17),13E-dien-15-acetate (2),were isolated fromthe needles of Pinus sylvestris.Their structures were elucidated by spectroscopic methods,including 2D-NMR spectra.Compound1 exhibited the significant eytotoxic activity against the human carcinoma cell lines Hela,SK-N-SH and BEL-7402 in vitro.

  3. Two diterpene rhamnosides, mimosasides B and C, from Mimosa hostilis.

    Science.gov (United States)

    Ohsaki, Ayumi; Yokoyama, Ritsuko; Miyatake, Haruka; Fukuyama, Yoshiyasu

    2006-12-01

    Two new diterpene rhamnosides, mimosasides B and C (1, 2) were isolated together with mimosaside A (3), a known diterpene rhamnoside (4), four known flavones (5-8), five known flavanones (9-13), and four known chalcones (14-17) from the leaves and twigs of a Brazilian medicinal plant, Mimosa hostilis.

  4. The Diterpene Sclareol Vascular Effect in Normotensive and Hypertensive Rats.

    Science.gov (United States)

    Campos, Debora Ribeiro; Celotto, Andrea Carla; Albuquerque, Agnes Afrodite S; Ferreira, Luciana Garros; Monteiro, Ariadne Santana E Neves; Coelho, Eduardo Barbosa; Evora, Paulo Roberto Barbosa

    2017-06-29

    The diterpene Sclareol has antimicrobial action, cytotoxic and cytostatic effects and anti-tumor activities. However, researches on the cardiovascular system are scarce. This study was designed to investigate the mechanisms involved in the Sclareol cardiovascular effect in normotensive and hypertensive rats. The arterial hypertension was promoted using 2-kidneys 1-clip model in rats. The effect of sclareol on blood pressure was performed by using three dose (10, 20 and 40 mg/kg). Cumulative dose-response curves for Sclareol were determined for endothelium-intact and endothelium-denuded aortic rings in presence or absence of L-NAME and ODQ. The NOx levels were measure in the plasma sample. The Sclareol administration in vivo caused a significant reduction in blood pressure in both groups. In vitro the sclareol promoted relaxation in aorta, with endothelium, pre-contracted to Phe. The inhibitors of the nitric oxide synthase and soluble guanylate cyclase were as efficient as the removal of endothelium, in inhibiting the Sclareol-induced relaxation. Otherwise, it was no change of NOx. Also, for unknown reasons, the Sclareol is not selective for hypertensive animals. The diterpene Sclareol showed in vivo hypotensive and in-vitro vasodilator effects; The chemiluminescence plasmatic NO analysis showed no significant difference between groups and The Sclareol exhibit better effect on normotensive than hypertensive animals to reduce blood pressure. It is concluded that the diterpenes metabolites would be a promising source prototype for the development of new agents in the cardiovascular therapy. O diterpeno Esclareol tem ação antimicrobiana, efeitos citotóxicos e citostáticos e atividades antitumorais. No entanto, pesquisas sobre o sistema cardiovascular são escassas. Este estudo foi desenvolvido para investigar os mecanismos envolvidos no efeito cardiovascular de Esclareol em ratos normotensos e hipertensos. A hipertensão arterial foi promovida utilizando modelo de

  5. The terpene synthase gene family in Tripterygium wilfordii harbors a labdane-type diterpene synthase among the monoterpene synthase TPS-b subfamily

    DEFF Research Database (Denmark)

    Hansen, Nikolaj L; Heskes, Allison M; Hamberger, Britta;

    2017-01-01

    in the formation of four C-20 diphosphate intermediates, precursors of both generalized and specialized metabolism and a novel scaffold for Celastraceae. Functional pairs of the class I and II enzymes resulted in formation of three scaffolds, accounting for some of the terpenoid diversity found in T. wilfordii...

  6. Synthesis and Antitumor Activity of Diterpenylhydroquinone Derivatives of Natural Ent-Labdanes

    Directory of Open Access Journals (Sweden)

    Mauricio Cuellar Fritis

    2010-09-01

    Full Text Available Two new compounds 2β-acetoxy-15-phenyl-(22,25-acetoxy-ent-labda-8(17, 13(E-diene (9 and 2β-hydroxy-15-phenyl-(22,24,26-trimethoxy-ent-labda-8(17,13(E-diene (10 have been prepared by an Electrophilic Aromatic Substitution (EAS reaction between diterpenyl allylic alcohols and 1,4-hydroquinone or 1,3,5-trimethoxybenzene using BF3.Et2O as a catalyst. These compounds, along with a series of natural ent-labdanes 3-8, have been evaluated for their in vitro cytotoxic activities against cultured human cancer cells of PC-3 and DU-145 human prostate cancer, MCF-7 and MDA-MB-231 breast carcinoma and dermal human fibroblasts (DHF. Some compounds displayed inhibition at µM IC50 values.

  7. Inhibition of Helicobacter pylori and Its Associate Urease by Labdane Diterpenoids Isolated from Andrographis paniculata.

    Science.gov (United States)

    Shaikh, Rafik U; Dawane, Ashwini A; Pawar, Rajendra P; Gond, Dhananjay S; Meshram, Rohan J; Gacche, Rajesh N

    2016-03-01

    The present study was carried out to evaluate anti-Helicobacter pylori and its associated urease activity of labdane diterpenoids isolated from Andrographis paniculata. A molecular docking analysis was performed by using ArgusLab 4.0.1 software. The results obtained indicate that compound A possesses strong inhibition to H. pylori, 28 ± 2.98 (minimum inhibitory concentration, 9 µg/mL), and its urease, 85.54 ± 2.62% (IC50 , 20.2 µg/mL). Compounds B, C, and D also showed moderate inhibition to H. pylori and its urease. The obtained results were in agreement with the molecular docking analysis of compounds. The phytochemicals under investigation were found to be promising antibacterial agents. Moreover, the isolated compounds can be considered as a resource for searching novel anti-H. pylori agents possessing urease inhibition.

  8. A New Diterpene Glycoside from Stevia rebaudiana

    Directory of Open Access Journals (Sweden)

    Indra Prakash

    2011-04-01

    Full Text Available From the commercial extract of the leaves of Stevia rebaudiana, a new diterpene glycoside was isolated besides the known steviol glycosides including stevioside, rebaudiosides A-F, rubusoside and dulcoside A. The new compound was identified as 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyloxy] ent-kaur-16-en-19-oic acid-(2-O-α-L-rhamnopyranosyl-β-D-glucopyranosyl ester (1 on the basis of extensive spectroscopic (NMR and MS and chemical studies.

  9. Asymmetric synthesis and absolute stereochemistry of a labdane-type diterpenoid isolated from the rhizomes of Isodan yuennanensis.

    Science.gov (United States)

    Deng, Heping; Cao, Wei; Zhang, Zhijiang; Liu, Bo

    2016-07-14

    The first synthesis of a labdane-type diterpenoid isolated from Isodon yuennanensis was achieved in fourteen steps from commercially available starting material, (+)-sclareolide. The synthesis features the Barton nitrite ester reaction to introduce an oxime at the angular methyl group and the Jones oxidation to construct the lactone segment. By comparison of the optical rotation of our synthetic sample and the natural sample, the absolute stereochemistry of the natural diterpenoid has been determined.

  10. Diterpene alkaloids and diterpenes from Spiraea japonica and their anti-tobacco mosaic virus activity.

    Science.gov (United States)

    Ma, Yuan; Mao, Xin-Ying; Huang, Lie-Jun; Fan, Yi-Min; Gu, Wei; Yan, Chen; Huang, Tao; Zhang, Jian-Xin; Yuan, Chun-Mao; Hao, Xiao-Jiang

    2016-03-01

    Five new naturally occurring natural products, including two atisine-type diterpene alkaloids (1 and 2), two atisane-type diterpenes (3 and 4), and a new natural product spiramine C2 (5), along with nine known ones (6-14), were isolated from the ethanolic extracts of the whole plant of Spiraea japonica var. acuminata Franch. Their structures were elucidated by extensive spectroscopic analysis. The anti-tobacco mosaic virus (TMV) activities of all the compounds were evaluated by the conventional half-leaf method. Six compounds (2, 3, 6, 7, 11, and 12) exhibited moderate activities at 100 μg/mL with inhibition rates in the range of 69.4-92.9%, which were higher than that of the positive control, ningnanmycin. Their preliminary structure-activity relationships were also discussed.

  11. Trocheliolide A, a Hydroperoxycembranoidal Diterpene from the Octocoral Sarcophyton trocheliophorum.

    Science.gov (United States)

    Liu, Kuan-Ming; Cheng, Ching-Hsiao; Chen, Wu-Fu; Lu, Mei-Chin; Fang, Lee-Shing; Wen, Zhi-Hong; Su, Jui-Hsin; Wu, Yang-Chang; Sung, Ping-Jyun

    2015-07-01

    A new hydroperoxycembranoidal diterpene, trocheliolide A (1), was isolated from the octocoral Sarcophyton trocheliophorum. The structure of 1 was elucidated on the basis of spectroscopic and mass spectrometric methods.

  12. Plant diterpene synthases: exploring modularity and metabolic diversity for bioengineering.

    Science.gov (United States)

    Zerbe, Philipp; Bohlmann, Jörg

    2015-07-01

    Plants produce thousands of diterpenoid natural products; some of which are of significant industrial value as biobased pharmaceuticals (taxol), fragrances (sclareol), food additives (steviosides), and commodity chemicals (diterpene resin acids). In nature, diterpene synthase (diTPS) enzymes are essential for generating diverse diterpene hydrocarbon scaffolds. While some diTPSs also form oxygenated compounds, more commonly, oxygenation is achieved by cytochrome P450-dependent mono-oxygenases. Recent genome-, transcriptome-, and metabolome-guided gene discovery and enzyme characterization identified novel diTPS functions that form the core of complex modular pathway systems. Insights into diterpene metabolism may translate into the development of new bioengineered microbial and plant-based production systems.

  13. Two new diterpene and sesquiterpene from Pteris dispar

    Institute of Scientific and Technical Information of China (English)

    Zhan Ping Gou; Nian Ci Liang; Jie Hou; Li Li; Yan Tian; Chang Yuan Wang; Xun Cui; Bao Jing Zhang; Xiao Chi Ma

    2011-01-01

    A new diterpene and a new sesquiterpene were isolated from Pteris dispar and identified on the basis of spectral data including 2D NMR. Compounds 1 and 2 showed potent cytotoxic activity against KB cell.

  14. Neo-clerodane diterpenes from the hallucinogenic sage Salvia divinorum.

    Science.gov (United States)

    Shirota, Osamu; Nagamatsu, Kumi; Sekita, Setsuko

    2006-12-01

    Seven new neo-clerodane diterpenes, salvidivins A (2), B, (3), C (4), and D (5), salvinorins H (6) and I (7), and divinatorin [corrected] F (8), along with eight known neo-clerodane diterpenes, salvinorins A (1)-F, divinatorins A and B, and seven other constituents, were isolated from the hallucinogenic sage Salvia divinorum. The structures of 1-7 were elucidated on the basis of 2D NMR spectroscopic studies.

  15. Anti-HIV diterpenes from Coleus forskohlii.

    Science.gov (United States)

    Bodiwala, Hardik S; Sabde, Sudeep; Mitra, Debashis; Bhutani, Kamlesh Kumar; Singh, Inder Pal

    2009-09-01

    Various extracts of the aerial parts of Coleus forskohlii (Labiatae) were prepared and evaluated at their non cytotoxic concentration against HIV-1 NL4-3. Chloroform, ethyl acetate and n-butanol extracts showed 45.6, 66.5 and 37.7% inhibition of HIV, respectively in CEM-GFP cells infected with HIV-1(NL4-3) at 5 microg/mL. Four diterpenes, 1-deoxyforskolin, 1,9-dideoxyforskolin, forskolin and isoforskolin were isolated from the chloroform extract and tested against the virus. Six semi-synthetic derivatives of forskolin have been prepared to study SAR. 1-Deoxyforskolin and forskolin were found to be active against HIV(NL4-3). This is first report of anti HIV activity of this plant and its isolated constituents.

  16. Synthesis of ent-Kaurane Diterpene Monoglycosides

    Directory of Open Access Journals (Sweden)

    Venkata Sai Prakash Chaturvedula

    2011-10-01

    Full Text Available Synthesis of two ent-kaurane diterpene glycosides, steviol 19-O-β-D-glucopyranosiduronic acid (steviol glucuronide, 5, and 13-hydroxy ent-kaur-16-en-19-oic acid-β-D-glucopyranosyl ester (7 has been achieved from a common starting material, steviol, using phase transfer catalyst. Also, synthesis of an additional 17-nor-ent-kaurane glycoside, namely 13-methyl-16-oxo-17-nor-ent-kauran-19-oic acid-β-D-glucopyranosyl ester (10 was performed using the starting material isosteviol and similar synthetic methodology. Synthesis of all three steviol glycosides was performed using straightforward chemistry and their structures were characterized on the basis of 1D and 2D NMR as well as mass spectral (MS data.

  17. Defensive Diterpene from the Aeolidoidean Phyllodesmium longicirrum.

    Science.gov (United States)

    Bogdanov, Alexander; Hertzer, Cora; Kehraus, Stefan; Nietzer, Samuel; Rohde, Sven; Schupp, Peter J; Wägele, Heike; König, Gabriele M

    2016-03-25

    Phyllodesmium is a tropical marine slug genus with about 30 described species. None of them have a protective shell, and all of them feed on octocorals that are generally known to provide defensive compounds and thus help to defend the naked slugs against sympatric predators, such as fish, crabs, cephalopods, and echinoderms. Phyllodesmium longicirrum is the species that grows the biggest and that is least protected by camouflage on its respective food, usually a soft coral of the genus Sarcophyton. Investigation of the lipophilic extract of a single specimen of P. longicirrum from the Great Barrier Reef (Australia) led to the isolation of four new polycyclic diterpenes. Compound 1 showed significant deterrent activity in a fish feeding assay.

  18. Trichodermaerin: a diterpene lactone from Trichoderma asperellum

    Directory of Open Access Journals (Sweden)

    Suchada Chantrapromma

    2014-04-01

    Full Text Available The title compound, C20H28O3, known as `trichodermaerin' [systematic name: (4E-4,9,15,16,16-pentamethyl-6-oxatetracyclo[10.3.1.01,10.05,9]hexadec-4-ene-7,13-dione], is a diterpene lactone which was isolated from Trichoderma asperellum. The structure has a tetracycic 6–5–7–5 ring system, with the cyclohexanone ring adopting a twisted half-chair conformation and the cyclopentane ring adopting a half-chair conformation, whereas the cycloheptene and tetrahydrofurananone rings are in chair and envelope (with the methyl-substituted C atom as the flap conformations, respectively. The three-dimensional architecture is stabilized by C—H...O interactions.

  19. Trichodermaerin: a diterpene lactone from Trichoderma asperellum

    Science.gov (United States)

    Chantrapromma, Suchada; Jeerapong, Chotika; Phupong, Worrapong; Quah, Ching Kheng; Fun, Hoong-Kun

    2014-01-01

    The title compound, C20H28O3, known as ‘trichodermaerin’ [systematic name: (4E)-4,9,15,16,16-penta­methyl-6-oxa­tetra­cyclo­[10.3.1.01,10.05,9]hexa­dec-4-ene-7,13-dione], is a diterpene lactone which was isolated from Trichoderma asperellum. The structure has a tetra­cycic 6–5–7–5 ring system, with the cyclo­hexa­none ring adopting a twisted half-chair conformation and the cyclo­pentane ring adopting a half-chair conformation, whereas the cyclo­heptene and tetra­hydro­furan­anone rings are in chair and envelope (with the methyl-substituted C atom as the flap) conformations, respectively. The three-dimensional architecture is stabilized by C—H⋯O inter­actions. PMID:24826124

  20. Diterpene Constituents of Euphorbia exigua L. and Multidrug Resistance Reversing Activity of the Isolated Diterpenes.

    Science.gov (United States)

    Rédei, Dóra; Boros, Klára; Forgo, Peter; Molnár, Joseph; Kele, Zoltán; Pálinkó, István; Pinke, Gyula; Hohmann, Judit

    2015-08-01

    Phytochemical investigation of the MeOH extract obtained from the aerial parts of the annual weed Euphorbia exigua L. resulted in the isolation of two novel (1, 2) and one known (3) jatrophane diterpenes. Their structures were established by extensive 1D- and 2D-NMR spectroscopy and HR-ESI-MS. The isolated compounds were evaluated for multidrug resistance (MDR) reversing activity on human MDR gene-transfected L5178 mouse lymphoma cells; and all three compounds were found to modulate the intracellular drug accumulation.

  1. Diterpene Cyclases and the Nature of the Isoprene Fold

    Science.gov (United States)

    Cao, Rong; Zhang, Yonghui; Mann, Francis M.; Huang, Cancan; Mukkamala, Dushyant; Hudock, Michael P.; Mead, Matthew; Prisic, Sladjana; Wang, Ke; Lin, Fu-Yang; Chang, Ting-Kai; Peters, Reuben; Oldfield, Eric

    2013-01-01

    The structures and mechanism of action of many terpene cyclases are known, but there are no structures of diterpene cyclases. Here, we propose structural models based on bioinformatics, site-directed mutagenesis, domain swapping, enzyme inhibition and spectroscopy that help explain the nature of diterpene cyclase structure, function, and evolution. Bacterial diterpene cyclases contain ∼20 α-helices and the same conserved “QW” and DxDD motifs as in triterpene cyclases, indicating the presence of a βγ barrel structure. Plant diterpene cyclases have a similar catalytic motif and βγ-domain structure together with a third, α-domain, forming an αβγ structure, and in H+-initiated cyclases, there is an EDxxD-like Mg2+/diphosphate binding motif located in the γ-domain. The results support a new view of terpene cyclase structure and function and suggest evolution from ancient (βγ) bacterial triterpene cyclases to (βγ) bacterial and thence to (αβγ) plant diterpene cyclases. PMID:20602361

  2. Interaction of coffee compounds with serum albumins. Part II: Diterpenes.

    Science.gov (United States)

    Guercia, Elena; Forzato, Cristina; Navarini, Luciano; Berti, Federico

    2016-05-15

    Cafestol and 16-O-methylcafestol are diterpenes present in coffee, but whilst cafestol is found in both Coffea canephora and Coffea arabica, 16-O-methylcafestol (16-OMC) was reported to be specific of only C. canephora. The interactions of such compounds, with serum albumins, have been studied. Three albumins have been considered, namely human serum albumin (HSA), fatty acid free HSA (ffHSA) and bovine serum albumin (BSA). The proteins interact with the diterpenes at the interface between Sudlow site I and the fatty acid binding site 6 in a very peculiar way, leading to a significant change in the secondary structure. The diterpenes do not displace reference binding drugs of site 2, but rather they enhance the affinity of the site for the drugs. They, therefore, may alter the pharmacokinetic profile of albumin - bound drugs.

  3. A new tigliane-type diterpene from Euphorbia dracunculoides Lam.

    Science.gov (United States)

    Dai, Lin-Feng; Liang, Qian; Liu, Tie; He, Ming-Yu; Zhao, Ping; Xu, Wen-Hui

    2016-07-01

    One new tigliane-type diterpene, 4-deoxy-4(β)H-8-hydroperoxyphorbol-12-benzoate-13-isobutyrate (1), together with two known diterpenoids, 3-acetyl-5,8-dibenzoyl-14α-propanoyl-13,17-epoxy-7-myrsinaone diterpene with C9-C10 cyclised to form an additional lactone ring (2), Euphodendriane A (3) have been isolated from the whole plants of Euphorbia dracunculoides Lam. Their structures were elucidated by means of extensive spectroscopic analysis (NMR and HR-ESI-MS) and comparison with data reported in the literature. This is the first isolation of 8-hydroperoxy tigliane diterpene (1) from the genus of Euphorbia. All compounds were evaluated for their antifungal activities.

  4. Phytotoxic clerodane diterpenes from Salvia miniata Fernald (Lamiaceae).

    Science.gov (United States)

    Bisio, Angela; Damonte, Gianluca; Fraternale, Daniele; Giacomelli, Emanuela; Salis, Annalisa; Romussi, Giovanni; Cafaggi, Sergio; Ricci, Donata; De Tommasi, Nunziatina

    2011-02-01

    Our ongoing research to identify natural growth inhibitors with diterpene and triterpene skeletons exuding from the surface of the aerial parts of Salvia species led us to study Salvia miniata Fernald. Ten clerodane diterpenoids were found, along with three known diterpenes. Most of the isolated compounds from S. miniata inhibited the germination of Papaver rhoeas L. and Avena sativa L. in Petri dish experiments. Parallel results have been obtained in experiments carried out to evaluate the subsequent growth of the seedlings of the target species in the presence of the tested compounds.

  5. Novel diterpenes from the heartwood of Chamaecyparis obtusa var. formosana.

    Science.gov (United States)

    Kuo, Yueh-Hsiung; Chen, Chia-Hsien; Chien, Shih-Chang; Lin, Hsiu-Chuan

    2004-06-01

    Two novel diterpenes, obtusanal B (1) and obtusadione (2), along with obtusanal A (3), obtunone (4), 12-hydroxy-6,7-secoabieta-8,11,13-triene-6,7-dial, 8,12-dihydroxydielmentha-5,9-diene-7,11-dione and myrcene, isolated from the heartwood of Chamaecyparis obtusa var. formosana, were characterized by spectroscopic means, including 2D-NMR techniques. Compounds 1 and 2 are 7(6-->2)abeoabietane and 14(8-->9)abeoabietane type diterpenes, respectively. Their biosyntheses were proposed.

  6. Methyl carnosate, an antibacterial diterpene isolated from Salvia officinalis leaves.

    Science.gov (United States)

    Climati, Elisa; Mastrogiovanni, Fabio; Valeri, Maria; Salvini, Laura; Bonechi, Claudia; Mamadalieva, Nilufar Zokirzhonovna; Egamberdieva, Dilfuza; Taddei, Anna Rita; Tiezzi, Antonio

    2013-04-01

    Ethanolic extracts of Salvia officinalis leaves demonstrated antibacterial activity against Bacillus cereus. Fractionation of the extracts led to the isolation of the most active antibacterial compound, which, from spectroscopic and LC-MS evidence, was proved to be the diterpene, methyl carnosate.

  7. Insect antifeedant activity of clerodane diterpenes and related model compounds

    NARCIS (Netherlands)

    Klein Gebbinck, E.A.; Jansen, B.J.M.; Groot, de Æ.

    2002-01-01

    A comprehensive compilation of all test results on the insect antifeedant activity of clerodane diterpenes and related model compounds is reported. To increase the compatibility of data from different sources, some of the results reported in the literature have been converted into a standardized for

  8. A New Diterpene from Suregada glomerulate(Blume) Baill

    Institute of Scientific and Technical Information of China (English)

    Su Li HE; Ruo Yun CHEN

    2005-01-01

    A new diterpene with an abietane skeleton, 3β-hydroxy-8α, 14α-epoxyabieta-11,13(15)-dien-16,12-olide, was isolated from the root of Suregada glomerulate(Blume) Baill. Its structure was elucidated on the basis of spectroscopic and X-ray crystallographic analysis.

  9. A new lathyrane diterpene glycoside from Euphorbia helioscopia L.

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A new lathyrane diterpene glycoside,named 3β,7β,15β-trihydroxy-14-oxolathyra-5E,12E-dienyl-16-O-β-D-glucopyranoside, was isolated from Euphorbia helioscopia L.Its structure was established by spectroscopic techniques including 2D NMR.

  10. Novel neoclerodane diterpene derivatives from the smoke of salvinorin A

    Science.gov (United States)

    Ma, Zhongze; Deng, Gang; Lee, David Y. W.

    2010-01-01

    Salvinorin A is a naturally-occurring potent and selective kappa opioid receptor agonist, and smoking salvinorin A produces the most intense hallucinogenic effects in human. Eight neoclerodane diterpene derivatives were isolated from the smoke of salvinorin A, and their structures were identified by spectroscopic methods. The major structural changes include epimerizations, eliminations, and rearrangements. PMID:20936100

  11. An Abietane Diterpene and a Sterol from Fungus Phellinus igniarius

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A new abietane diterpene 12-hydroxy-7-oxo-5, 8, 11, 13-tetraene-18, 6-abietanolide,together with a new natural sterol stigmasta-7, 22-diene-3β, 5α, 6α-triol have been isolated from the fruiting body of the fungus Phellinus igniarius. Their structures were elucidated by spectroscopic methods including 2D NMR techniques.

  12. 香茶菜属植物中二萜类化合物的研究进展%Study and Progress of Diterpenes of Rabdosia

    Institute of Scientific and Technical Information of China (English)

    张小清

    2003-01-01

    It is confirmed the diterpenes is possessed of these reaction activitor、 acidylationoxidation、 disoxidate by analyzing theconstruction of diterpenes. Then the diterpenes of Rabdosia possess a wide range of physiology activitor mainly in anicancer、antibiotic.

  13. Evolution of Conifer Diterpene Synthases: Diterpene Resin Acid Biosynthesis in Lodgepole Pine and Jack Pine Involves Monofunctional and Bifunctional Diterpene Synthases1[W][OA

    Science.gov (United States)

    Hall, Dawn E.; Zerbe, Philipp; Jancsik, Sharon; Quesada, Alfonso Lara; Dullat, Harpreet; Madilao, Lina L.; Yuen, Macaire; Bohlmann, Jörg

    2013-01-01

    Diterpene resin acids (DRAs) are major components of pine (Pinus spp.) oleoresin. They play critical roles in conifer defense against insects and pathogens and as a renewable resource for industrial bioproducts. The core structures of DRAs are formed in secondary (i.e. specialized) metabolism via cycloisomerization of geranylgeranyl diphosphate (GGPP) by diterpene synthases (diTPSs). Previously described gymnosperm diTPSs of DRA biosynthesis are bifunctional enzymes that catalyze the initial bicyclization of GGPP followed by rearrangement of a (+)-copalyl diphosphate intermediate at two discrete class II and class I active sites. In contrast, similar diterpenes of gibberellin primary (i.e. general) metabolism are produced by the consecutive activity of two monofunctional class II and class I diTPSs. Using high-throughput transcriptome sequencing, we discovered 11 diTPS from jack pine (Pinus banksiana) and lodgepole pine (Pinus contorta). Three of these were orthologous to known conifer bifunctional levopimaradiene/abietadiene synthases. Surprisingly, two sets of orthologous PbdiTPSs and PcdiTPSs were monofunctional class I enzymes that lacked functional class II active sites and converted (+)-copalyl diphosphate, but not GGPP, into isopimaradiene and pimaradiene as major products. Diterpene profiles and transcriptome sequences of lodgepole pine and jack pine are consistent with roles for these diTPSs in DRA biosynthesis. The monofunctional class I diTPSs of DRA biosynthesis form a new clade within the gymnosperm-specific TPS-d3 subfamily that evolved from bifunctional diTPS rather than monofunctional enzymes (TPS-c and TPS-e) of gibberellin metabolism. Homology modeling suggested alterations in the class I active site that may have contributed to their functional specialization relative to other conifer diTPSs. PMID:23370714

  14. Biosynthesis of uterotonic diterpenes from Montanoa tomentosa (zoapatle).

    Science.gov (United States)

    Villa-Ruano, Nemesio; Betancourt-Jiménez, Martha Guadalupe; Lozoya-Gloria, Edmundo

    2009-12-15

    Montanoa tomentosa (zoapatle) is a Central American plant used in Mexico in traditional herbal medicine to ease childbirth labor and to cure certain female disorders. Recently, crude extracts of M. tomentosa have been reported to have an aphrodisiacal effect on male rats. The bioactive molecules are the uterotonic diterpenes kaurenoic acid (KA), grandiflorenic acid (GF), and monoginoic acid (MO). Roots of M. tomentosa contain all three diterpenes, whereas in leaves only kaurenoic and GF are present. However, despite the pharmacological importance of these compounds, specific information about their biosynthesis and localization in the plant is not available. In this investigation, we followed the metabolic transformation of a tritium-labeled diterpene-precursor via geranylgeranyl diphosphate into each of the three diterpenes. Inhibitors of gibberellin biosynthesis were used to elucidate the sequence of conversion of the intermediates. Our results suggest the biosynthetic conversion of KA into GF by a putative cytochrome P450-like desaturase. Partial characterization of the enzyme revealed that it requires NADPH and O2 but is inhibited by 50 microM paclobutrazol, suggesting a cytochrome P450 desaturase like enzyme (EC 1.14.14.-). Optimal reaction conditions are 32 degrees C and a pH of 7.6, respectively. Apparent kinetics parameters for KA gave a K(m,app) of 36.31 microM, and a V(max, app) of 13.6 nmol KA mg(1)protein h(-1). Based on the data presented, a putative biosynthetic pathway is proposed for the uterotonic diterpenes of M. tomentosa.

  15. Biological Activities and Cytotoxicity of Diterpenes from Copaifera spp. Oleoresins

    Directory of Open Access Journals (Sweden)

    Fabiano de S. Vargas

    2015-04-01

    Full Text Available Copaifera spp. are Amazonian species widely studied and whose oleoresins are used by local people for various medicinal purposes. However, a detailed study of the activity of the main phytochemical components of these oleoresins remains to be done. Here, we studied the cytotoxicity and in vitro anti-inflammatory effects of six diterpene acids: copalic, 3-hydroxy-copalic, 3-acetoxy-copalic, hardwickiic, kolavic-15-metyl ester, and kaurenoic, isolated from the oleoresins of Copaifera spp. The diterpenes did not show cytotoxicity in normal cell lines, nor did they show significant changes in viability of tumoral line cells. The 3-hydroxy-copalic was able to inhibit the enzyme tyrosinase (64% ± 1.5% at 250 µM. The kolavic-15-metyl ester at 200 µM showed high inhibitory effect on lipoxygenase (89.5% ± 1.2%. Among the diterpenes tested, only kaurenoic and copalic acids showed significant hemolytic activities with 61.7% and 38.4% at 100 µM, respectively. In addition, it was observed that only the copalic acid (98.5% ± 1.3% and hardwickiic acid (92.7% ± 4.9% at 100 mM inhibited nitric oxide production in macrophages activated by lipopolysaccharide. In this assay, the diterpenes did not inhibit tumor necrosis factor-α production. The acids inhibited the production of IL-6, 3-acetoxy-copalic (23.8% ± 8.2%, kaurenoic (11.2% ± 5.7%, kolavic-15-methyl ester (17.3% ± 4.2%, and copalic (4.2% ± 1.8%, respectively, at 25 µM. The kaurenoic, 3-acetoxy-copalic and copalic acids increased IL-10 production. This study may provide a basis for future studies on the therapeutic role of diterpenic acids in treating acute injuries such as inflammation or skin disorders.

  16. Biological activities and cytotoxicity of diterpenes from Copaifera spp. Oleoresins.

    Science.gov (United States)

    de S Vargas, Fabiano; D O de Almeida, Patrícia; Aranha, Elenn Suzany P; de A Boleti, Ana Paula; Newton, Peter; de Vasconcellos, Marne C; Junior, Valdir F Veiga; Lima, Emerson S

    2015-04-09

    Copaifera spp. are Amazonian species widely studied and whose oleoresins are used by local people for various medicinal purposes. However, a detailed study of the activity of the main phytochemical components of these oleoresins remains to be done. Here, we studied the cytotoxicity and in vitro anti-inflammatory effects of six diterpene acids: copalic, 3-hydroxy-copalic, 3-acetoxy-copalic, hardwickiic, kolavic-15-metyl ester, and kaurenoic, isolated from the oleoresins of Copaifera spp. The diterpenes did not show cytotoxicity in normal cell lines, nor did they show significant changes in viability of tumoral line cells. The 3-hydroxy-copalic was able to inhibit the enzyme tyrosinase (64% ± 1.5%) at 250 µM. The kolavic-15-metyl ester at 200 µM showed high inhibitory effect on lipoxygenase (89.5% ± 1.2%). Among the diterpenes tested, only kaurenoic and copalic acids showed significant hemolytic activities with 61.7% and 38.4% at 100 µM, respectively. In addition, it was observed that only the copalic acid (98.5% ± 1.3%) and hardwickiic acid (92.7% ± 4.9%) at 100 mM inhibited nitric oxide production in macrophages activated by lipopolysaccharide. In this assay, the diterpenes did not inhibit tumor necrosis factor-α production. The acids inhibited the production of IL-6, 3-acetoxy-copalic (23.8% ± 8.2%), kaurenoic (11.2% ± 5.7%), kolavic-15-methyl ester (17.3% ± 4.2%), and copalic (4.2% ± 1.8%), respectively, at 25 µM. The kaurenoic, 3-acetoxy-copalic and copalic acids increased IL-10 production. This study may provide a basis for future studies on the therapeutic role of diterpenic acids in treating acute injuries such as inflammation or skin disorders.

  17. Exploring diterpene metabolism in non-model species: transcriptome-enabled discovery and functional characterization of labda-7,13E-dienyl diphosphate synthase from Grindelia robusta.

    Science.gov (United States)

    Zerbe, Philipp; Rodriguez, Selina M; Mafu, Sibongile; Chiang, Angela; Sandhu, Harpreet K; O'Neil-Johnson, Mark; Starks, Courtney M; Bohlmann, Jörg

    2015-09-01

    Grindelia robusta or gumweed, is a medicinal herb of the sunflower family that forms a diverse suite of diterpenoid natural products. Its major constituents, grindelic acid and related grindelane diterpenoids accumulate in a resinous exudate covering the plants' surfaces, most prominently the unopened composite flower. Recent studies demonstrated potential pharmaceutical applications for grindelic acid and its synthetic derivatives. Mining of the previously published transcriptome of G. robusta flower tissue identified two additional diterpene synthases (diTPSs). We report the in vitro and in vivo functional characterization of an ent-kaurene synthase of general metabolism (GrTPS4) and a class II diTPS (GrTPS2) of specialized metabolism that converts geranylgeranyl diphosphate (GGPP) into labda-7,13E-dienyl diphosphate as verified by nuclear magnetic resonance (NMR) analysis. Tissue-specific transcript abundance of GrTPS2 in leaves and flowers accompanied by the presence of an endocyclic 7,13 double bond in labda-7,13E-dienyl diphosphate suggest that GrTPS2 catalyzes the first committed reaction in the biosynthesis of grindelic acid and related grindelane metabolites. With the formation of labda-7,13E-dienyl diphosphate, GrTPS2 adds an additional function to the portfolio of monofunctional class II diTPSs, which catalytically most closely resembles the bifunctional labda-7,13E-dien-15-ol synthase of the lycopod Selaginella moellendorffii. Together with a recently identified functional diTPS pair of G. robusta producing manoyl oxide, GrTPS2 lays the biosynthetic foundation of the diverse array of labdane-related diterpenoids in the genus Grindelia. Knowledge of these natural diterpenoid metabolic pathways paves the way for developing biotechnology approaches toward producing grindelic acid and related bioproducts.

  18. Expanding the landscape of diterpene structural diversity by stereochemically controlled combinatorial biosynthesis

    DEFF Research Database (Denmark)

    Andersen-Ranberg, Johan; Kongstad, Kenneth Thermann; Nielsen, Morten Thrane;

    2016-01-01

    Plant derived diterpenoids are relevant as pharmaceuticals, food additives and fragrances, yet a broader industrial utilization of these bioproducts is limited due to their low natural abundance and high structural complexity. Mimicking the modularity of diterpene biosynthesis in plants, we...... constructed 51 functional diterpene synthase combinations, 41 of which were “new-to-nature”. Here, we demonstrate stereoselective biosynthesis in Nicotiana benthamiana of 47 diterpene skeletons including natural variants and novel enantiomeric or diastereomeric counterparts. Scalable biotechnological...

  19. Gene Discovery of Modular Diterpene Metabolism in Nonmodel Systems1[W][OA

    Science.gov (United States)

    Zerbe, Philipp; Hamberger, Björn; Yuen, Macaire M.S.; Chiang, Angela; Sandhu, Harpreet K.; Madilao, Lina L.; Nguyen, Anh; Hamberger, Britta; Bach, Søren Spanner; Bohlmann, Jörg

    2013-01-01

    Plants produce over 10,000 different diterpenes of specialized (secondary) metabolism, and fewer diterpenes of general (primary) metabolism. Specialized diterpenes may have functions in ecological interactions of plants with other organisms and also benefit humanity as pharmaceuticals, fragrances, resins, and other industrial bioproducts. Examples of high-value diterpenes are taxol and forskolin pharmaceuticals or ambroxide fragrances. Yields and purity of diterpenes obtained from natural sources or by chemical synthesis are often insufficient for large-volume or high-end applications. Improvement of agricultural or biotechnological diterpene production requires knowledge of biosynthetic genes and enzymes. However, specialized diterpene pathways are extremely diverse across the plant kingdom, and most specialized diterpenes are taxonomically restricted to a few plant species, genera, or families. Consequently, there is no single reference system to guide gene discovery and rapid annotation of specialized diterpene pathways. Functional diversification of genes and plasticity of enzyme functions of these pathways further complicate correct annotation. To address this challenge, we used a set of 10 different plant species to develop a general strategy for diterpene gene discovery in nonmodel systems. The approach combines metabolite-guided transcriptome resources, custom diterpene synthase (diTPS) and cytochrome P450 reference gene databases, phylogenies, and, as shown for select diTPSs, single and coupled enzyme assays using microbial and plant expression systems. In the 10 species, we identified 46 new diTPS candidates and over 400 putatively terpenoid-related P450s in a resource of nearly 1 million predicted transcripts of diterpene-accumulating tissues. Phylogenetic patterns of lineage-specific blooms of genes guided functional characterization. PMID:23613273

  20. Synthesis and antiproliferative activity of new tonantzitlolone-derived diterpene derivatives.

    Science.gov (United States)

    Busch, Torsten; Dräger, Gerald; Kunst, Eike; Benson, Hannah; Sasse, Florenz; Siems, Karsten; Kirschning, Andreas

    2016-10-14

    The synthesis of the diterpene (+)-tonantzitlolone A and a series of derivatives is reported. The study includes the determination of their antiproliferative activities against selected cancer cell lines.

  1. Synthesis of Neoclerodane Diterpenes and Their Pharmacological Effects

    Science.gov (United States)

    Lovell, Kimberly M.; Prevatt-Smith, Katherine M.; Lozama, Anthony; Prisinzano, Thomas E.

    Salvinorin A is a neoclerodane diterpene that has been shown to be an agonist at kappa opioid receptors. Its unique structure makes it an attractive target for synthetic organic chemists due to its seven chiral centers and diterpene scaffold. This molecule is also interesting to pharmacologists because it is a non-serotonergic hallucinogen, and the first opioid ligand discovered that lacks a basic nitrogen. There have been several total synthesis approaches to salvinorin A, and these will be detailed within this chapter. Additionally, research efforts have concentrated on structure modification of the salvinorin A scaffold through semi-synthetic methods. Most modifications have focused on the manipulation of the acetate at C-2 and the furan ring. However, chemistry has also been developed to generate analogs at the C-1 ketone, the C-4 methyl ester, and the C-17 lactone. The synthetic methodologies developed for the salvinorin A scaffold will be described, as well as specific analogs with interesting biological activities.

  2. Antimicrobial Activity of Diterpenes from Viguiera arenaria against Endodontic Bacteria

    Directory of Open Access Journals (Sweden)

    Carlos H. G. Martins

    2011-01-01

    Full Text Available Six pimarane-type diterpenes isolated from Viguiera arenaria Baker and two semi-synthetic derivatives were evaluated in vitro against a panel of representative microorganisms responsible for dental root canal infections. The microdilution method was used for the determination of the minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC against Porphyromonas gingivalis, Prevotella nigrescens, Prevotella intermedia, Prevotella buccae, Fusobacterium nucleatum, Bacteroides fragilis, Actinomyces naeslundii, Actinomyces viscosus, Peptostreptococcus micros, Enterococcus faecalis and Aggregatibacter actinomycetemcomitans. The compounds ent-pimara-8(14,15-dien-19-oic acid, its sodium salt and ent-8(14,15-pimaradien-3β-ol were the most active, displaying MIC values ranging from 1 to 10 μg mL-1. The results also allow us to conclude that minor structural differences among these diterpenes significantly influence their antimicrobial activity, bringing new perspectives to the discovery of new chemicals for use as a complement to instrumental endodontic procedures.

  3. New pimarane diterpenes and other antimycobacterial metabolites from Anisochilus verticillatus.

    Science.gov (United States)

    Kulkarni, Roshan R; Shurpali, Ketaki; Khedkar, Vijay M; Puranik, Vedavati G; Sarkar, Dhiman; Joshi, Swati P

    2016-01-01

    Phytochemical investigation of the acetone extract of the aerial parts of Anisochilus verticillatus afforded a new 8,9-secopimarane diterpene (1), two new isopimarane diterpenes (2, 3) and the known ursolic acid (4), α-amyrin (5), β-amyrin (6), stigmast-5-en-3-one (7) and hydroxychavicol (8). Structures of the new compounds were elucidated with the help of 1D and 2D nuclear magnetic resonance spectroscopic data, and single crystal X-ray crystallography of compound 3. Compounds 2 and 8 inhibited Mycobacterium tuberculosis H37Ra with an IC50 of 11.3 (IC90 of 20.0 μg/mL) and 12.5 μg/mL, respectively. Correspondingly, molecular docking studies with Extra Precision Glide revealed a correlation between score and biological activity for these compounds to describe the molecular basis for the most significant SAR results.

  4. Two new diterpenes from the seeds of Caesalpinia minax Hance.

    Science.gov (United States)

    Lian, Lian; Li, Xiao-Bin; Yuan, Jiu-Zhi; Cheng, Li; Wu, Zhao-Hua; Gao, Hui-Yuan

    2015-01-01

    Molecules with diterpene skeletons often possess valuable medicinal properties. Two new diterpenes 1α,6α,7β-triacetoxy-5α-hydroxy-14β-ethyl-O-vouacapane (1) and 2α-acetoxy-14,15-cyclopimara-7β,16-diol (2) were isolated from the seeds of Caesalpinia minax Hance. Their structures were established on the basis of extensive spectroscopic analyses, including HR-ESI-MS, UV, IR, 1D, and 2D NMR (HSQC, HMBC, NOESY) methods. The stereochemical structure of 1 was confirmed via the circular dichroism spectrum and calculated ECD experiment. The inhibitory activity of nitric oxide production of RAW264.7 macrophages stimulated by lipopolysaccharide of compounds 1 and 2 was evaluated, and compound 1 was found to show significant inhibitory effect.

  5. Salvinicins A and B, new neoclerodane diterpenes from Salvia divinorum.

    Science.gov (United States)

    Harding, Wayne W; Tidgewell, Kevin; Schmidt, Matthew; Shah, Kushal; Dersch, Christina M; Snyder, John; Parrish, Damon; Deschamps, Jeffrey R; Rothman, Richard B; Prisinzano, Thomas E

    2005-07-07

    [reaction: see text] Two new neoclerodane diterpenes, salvinicins A (4) and B (5), were isolated from the dried leaves of Salvia divinorum. The structures of these compounds were elucidated by spectroscopic techniques, including (1)H and (13)C NMR, NOESY, HMQC, and HMBC. The absolute stereochemistry of these compounds was assigned on the basis of single-crystal X-ray crystallographic analysis of salvinicin A (4) and a 3,4-dichlorobenzoate derivative of salvinorin B.

  6. Antimalarial diterpene alkaloids from the seeds of Caesalpinia minax.

    Science.gov (United States)

    Ma, Guoxu; Sun, Zhaocui; Sun, Zhonghao; Yuan, Jingquan; Wei, Hua; Yang, Junshan; Wu, Haifeng; Xu, Xudong

    2014-06-01

    Two new diterpene alkaloids, caesalminines A (1) and B (2), possessing a tetracyclic cassane-type furanoditerpenoid skeleton with γ-lactam ring, were isolated from the seeds of Caesalpinia minax. Their structures were determined by different spectroscopic methods and ECD calculation. The plausible biosynthetic pathway of caesalminines A and B was proposed. The anti-malarial activity of compounds 1 and 2 is presented with IC50 values of 0.42 and 0.79 μM, respectively.

  7. Cassane diterpenes from the seed kernels of Caesalpinia sappan.

    Science.gov (United States)

    Nguyen, Hai Xuan; Nguyen, Nhan Trung; Dang, Phu Hoang; Thi Ho, Phuoc; Nguyen, Mai Thanh Thi; Van Can, Mao; Dibwe, Dya Fita; Ueda, Jun-Ya; Awale, Suresh

    2016-02-01

    Eight structurally diverse cassane diterpenes named tomocins A-H were isolated from the seed kernels of Vietnamese Caesalpinia sappan Linn. Their structures were determined by extensive NMR and CD spectroscopic analysis. Among the isolated compounds, tomocin A, phanginin A, F, and H exhibited mild preferential cytotoxicity against PANC-1 human pancreatic cancer cells under nutrition-deprived condition without causing toxicity in normal nutrient-rich conditions.

  8. Tasumatrols U-Z, taxane diterpene esters from Taxus sumatrana.

    Science.gov (United States)

    Shen, Ya-Ching; Wang, Shih-Sheng; Chien, Ching-Te; Kuo, Yao-Haur; Khalil, Ashraf Taha

    2008-04-01

    Phytochemical investigation of the leaves and twigs of Taxus sumatrana afforded six new taxane diterpene esters, tasumatrols U-Z ( 1- 6). Compounds 2 and 5 contained a rare five-membered lactone ring at C-8, C-9, C-10, and C-19. The structures were established on the basis of detailed spectroscopic analyses, particularly HRESIMS and 2D NMR techniques. Compound 5 showed cytotoxicity against a human hepatoma (Hep2) cell line.

  9. A New Cytotoxic Clerodane Diterpene from Casearia graveolens Twigs.

    Science.gov (United States)

    Meesakul, Pornphimol; Ritthiwigrom, Thunwadee; Cheenpracha, Sarot; Sripisut, Tawanun; Maneerat, Wisanu; Machan, Theeraphan; Laphookhieo, Surat

    2016-01-01

    The first phytochemical investigation of Casearia graveolens twigs led to the isolation and identification of a new clerodane diterpene, caseariagraveolin (1), together with six known compounds (2-7). Their structures were elucidated by intensive analysis of their spectroscopic data. Compound 1 showed strong cytotoxicity against oral cavity and breast cancer cell lines with IC₅₀ values of 2.48 and 6.63 µM, respectively.

  10. Trachylobane and kaurane diterpenes from Croton floribundus spreng

    Energy Technology Data Exchange (ETDEWEB)

    Uchoa, Paula Karina S.; Silva Junior, Jose Nunes da; Silveira, Edilberto Rocha; Lima, Mary Anne S., E-mail: mary@dqoi.ufc.br [Departamento de Quimica Organica e Inorganica, Centro de Ciencias, Universidade Federal do Ceara, Fortaleza, CE (Brazil); Braz-Filho, Raimundo [Laboratorio de Ciencias Quimicas, Centro de Ciencias Tecnologicas, Universidade Estadual do Norte Fluminense, Campo dos Goytacazes, RJ (Brazil); Costa-Lotufo, Leticia Veras; Araujo, Ana Jersia; Moraes, Manoel Odorico de; Pessoa, Claudia do O [Departamento de Farmacologia e Fisiologia, Centro de Ciencias da Saude, Universidade Federal do Ceara, Fortaleza, CE (Brazil)

    2013-09-01

    A new trachylobane diterpene ent-trachyloban-18,19-diol (1) was isolated from root bark of Croton floribundus, along with known diterpenes ent-trachyloban-19-oic acid (2), 15{beta}-hydroxy-ent-trachyloban-19-oic acid (3), ent-trachyloban-19-ol (4), ent-kaur-16-en-19-oic acid (5), ent-kaur-16-ene-6{alpha},19-diol (6) and ent-16{alpha}-hydroxykaur-11-en-19-oic acid (7). ent-trachyloban-18,19-diol (1) was submitted to derivatization reactions affording four new compounds (8-11). Cytotoxic activity of diterpenes 1, 3, 4, 7-11 against three human cancer cell lines was evaluated. No compounds showed cytotoxic potential with IC{sub 50} values greater than 25 {mu}g/mL. Compound 6 was evaluated against five human cancer cell lines, showing moderate effect against three cancer cell lines, MDA-MB-435, HCT-8 and HCT-116, with IC{sub 50} values of 14.32, 13.47 and 12.1 {mu}g/mL, respectively. (author)

  11. Volatile diterpene emission from dominant conifers in Japan

    Directory of Open Access Journals (Sweden)

    S. N. Matsunaga

    2011-07-01

    Full Text Available Reactive volatile organic compounds (VOCs are known to affect atmospheric chemistry, especially biogenic VOCs (BVOCs which have a significant impact on regional air quality due to their large emission rates and high reactivities. Diterpenes (Kaur-16-ene and a diterpene likely to be Hibaene were detected in all of the 205 enclosure air samples collected over multiple seasons at two different sites from Cryptomeria japonica and Chamaecyparis obtusa trees, the dominant coniferous trees in Japan. The emission rate of Kaur-16-ene, was determined to be from 0.01 to 7.1 μg dwg−1 h−1 (average: 0.61 μg dwg−1 h−1 by branch enclosure measurements and solid phase – liquid extraction techniques. The emission rate was an order of magnitude higher than that of monoterpenes which are known to comprise a major portion of total BVOC emissions. In addition, total emission of Kaur-16-ene at 30 °C was estimated to be comparable to that of total anthropogenic VOC emissions. To our knowledge, this is the first report of volatile diterpene emissions from trees.

  12. Volatile diterpene emission from dominant conifers in Japan

    Science.gov (United States)

    Matsunaga, S. N.; Chatani, S.; Nakatsuka, S.; Kusumoto, D.; Kubota, K.; Utsumi, Y.; Enoki, T.; Tani, A.; Hiura, T.

    2011-07-01

    Reactive volatile organic compounds (VOCs) are known to affect atmospheric chemistry, especially biogenic VOCs (BVOCs) which have a significant impact on regional air quality due to their large emission rates and high reactivities. Diterpenes (Kaur-16-ene and a diterpene likely to be Hibaene) were detected in all of the 205 enclosure air samples collected over multiple seasons at two different sites from Cryptomeria japonica and Chamaecyparis obtusa trees, the dominant coniferous trees in Japan. The emission rate of Kaur-16-ene, was determined to be from 0.01 to 7.1 μg dwg-1 h-1 (average: 0.61 μg dwg-1 h-1) by branch enclosure measurements and solid phase - liquid extraction techniques. The emission rate was an order of magnitude higher than that of monoterpenes which are known to comprise a major portion of total BVOC emissions. In addition, total emission of Kaur-16-ene at 30 °C was estimated to be comparable to that of total anthropogenic VOC emissions. To our knowledge, this is the first report of volatile diterpene emissions from trees.

  13. Trachylobane and kaurane diterpenes from Croton floribundus spreng

    Directory of Open Access Journals (Sweden)

    Paula Karina S. Uchôa

    2013-01-01

    Full Text Available A new trachylobane diterpene ent-trachyloban-18,19-diol (1 was isolated from root bark of Croton floribundus, along with known diterpenes ent-trachyloban-19-oic acid (2, 15b-hydroxy-ent-trachyloban-19-oic acid (3, ent-trachyloban-19-ol (4, ent-kaur-16-en-19-oic acid (5, ent-kaur-16-ene-6a,19-diol (6 and ent-16a-hydroxykaur-11-en-19-oic acid (7. ent-trachyloban-18,19-diol (1 was submitted to derivatization reactions affording four new compounds (8-11. Cytotoxic activity of diterpenes 1, 3, 4, 7-11 against three human cancer cell lines was evaluated. No compounds showed cytotoxic potential with IC50 values greater than 25 mg/mL. Compound 6 was evaluated against five human cancer cell lines, showing moderate effect against three cancer cell lines, MDA-MB-435, HCT-8 and HCT-116, with IC50 values of 14.32, 13.47 and 12.1 mg/mL, respectively.

  14. Antileishmanial activity of diterpene acids in copaiba oil

    Directory of Open Access Journals (Sweden)

    Adriana Oliveira dos Santos

    2013-02-01

    Full Text Available Leishmaniasis is a neglected tropical disease. According to the World Health Organization, there are approximately 1.5-two million new cases of cutaneous leishmaniasis each year worldwide. Chemotherapy against leishmaniasis is based on pentavalent antimonials, which were developed more than a century ago. The goals of this study were to investigate the antileishmanial activity of diterpene acids in copaiba oil, as well as some possible targets of their action against Leishmania amazonensis. Methyl copalate and agathic, hydroxycopalic, kaurenoic, pinifolic and polyaltic acids isolated from Copaifera officinales oleoresins were utilised. Ultrastructural changes and the specific organelle targets of diterpenes were investigated with electron microscopy and flow cytometry, respectively. All compounds had some level of activity against L. amazonensis. Hydroxycopalic acid and methyl copalate demonstrated the most activity against promastigotes and had 50% inhibitory concentration (IC50 values of 2.5 and 6.0 µg/mL, respectively. However, pinifolic and kaurenoic acid demonstrated the most activity against axenic amastigote and had IC50 values of 3.5 and 4.0 µg/mL, respectively. Agathic, kaurenoic and pinifolic acid caused significant increases in plasma membrane permeability and mitochondrial membrane depolarisation of the protozoan. In conclusion, copaiba oil and its diterpene acids should be explored for the development of new antileishmanial drugs.

  15. Antileishmanial activity of diterpene acids in copaiba oil

    Science.gov (United States)

    dos Santos, Adriana Oliveira; Izumi, Erika; Ueda-Nakamura, Tânia; Dias-Filho, Benedito Prado; da Veiga-Júnior, Valdir Florêncio; Nakamura, Celso Vataru

    2013-01-01

    Leishmaniasis is a neglected tropical disease. According to the World Health Organization, there are approximately 1.5-two million new cases of cutaneous leishmaniasis each year worldwide. Chemotherapy against leishmaniasis is based on pentavalent antimonials, which were developed more than a century ago. The goals of this study were to investigate the antileishmanial activity of diterpene acids in copaiba oil, as well as some possible targets of their action against Leishmania amazonensis. Methyl copalate and agathic, hydroxycopalic, kaurenoic, pinifolic and polyaltic acids isolated from Copaifera officinales oleoresins were utilised. Ultrastructural changes and the specific organelle targets of diterpenes were investigated with electron microscopy and flow cytometry, respectively. All compounds had some level of activity against L. amazonensis. Hydroxycopalic acid and methyl copalate demonstrated the most activity against promastigotes and had 50% inhibitory concentration (IC50) values of 2.5 and 6.0 µg/mL, respectively. However, pinifolic and kaurenoic acid demonstrated the most activity against axenic amastigote and had IC50 values of 3.5 and 4.0 µg/mL, respectively. Agathic, kaurenoic and pinifolic acid caused significant increases in plasma membrane permeability and mitochondrial membrane depolarisation of the protozoan. In conclusion, copaiba oil and its diterpene acids should be explored for the development of new antileishmanial drugs. PMID:23440116

  16. Cloning of Casbene Synthases from Euphorbiaceae Plants and Implications for Diterpene Biosynthesis in This Family

    Science.gov (United States)

    A large number of diterpenes have been isolated from Euphorbiaceae plants, many of which are of interest due to toxicity or potential therapeutic activity. Specific Euphorbiaceae diterpenes of medical interest include the latent HIV-1 activator prostratin (and related 12-deoxyphorbol esters), the an...

  17. Clerodane and Ent-kaurane Diterpene Glycosyl and Glycoside Derivatives from the Leaves of Casearia sylvestris

    Science.gov (United States)

    Five new clerodane diterpene glycosides caseariasides A-E (1-4) and three new ent-kaurane diterpene glucosides sylvestrisides C-E (6-8) were isolated from the leaves of Casearia sylvestris. Their structures were determined on the basis of chemical and spectroscopic analyses....

  18. New adduct of abietane-type diterpene from Salvia leriifolia Benth.

    Science.gov (United States)

    Hussain, Amjad; Adhikari, Achyut; Iqbal Choudhary, M; Ayatollahi, Syed Abdulmajid; Atta-Ur-Rahman

    2016-07-01

    A new adduct of abietane-type diterpene, salvialeriicone (1), was isolated from Salvia leriifolia Benth., along with a new chemical entity nor-abietane diterpene, 2-isopropyl-8,8-dimethyl-7,8-dihydrophenanthrene-1,4,5(6H)-trione (2). Their structures were determined using mass spectrometry, and 1D- and 2D-NMR spectroscopy.

  19. New Cembranoid Diterpenes from the Cultured Octocoral Nephthea columnaris

    Directory of Open Access Journals (Sweden)

    Ting-Hsi Hsiao

    2015-07-01

    Full Text Available Two new 15-hydroxycembranoid diterpenes, 2β-hydroxy-7β,8α-epoxynephthenol (1 and 2β-hydroxy-11α,12β-epoxynephthenol (2, were isolated from extracts of the octocoral Nephthea columnaris along with a new natural cembrane, epoxynephthenol (3 and a known sterol, nephalsterol A (4. The structures of cembranes 1–3 were elucidated by spectroscopic methods and comparison of the spectroscopic data with those of related analogues. The cytotoxicity of metabolites 1–4 against a panel of tumor cells is also described.

  20. A Diterpene Endoperoxide from Microtoena insuavis (Hance) Prain ex Dunn

    Institute of Scientific and Technical Information of China (English)

    Chong-Qian Li; Bo-Gang Li; Feng-Peng Wang; Guo-Lin Zhang

    2006-01-01

    A novel endoperoxide diterpene, 7α-hydroxy-abieta-8(14)-en-18-ol 9α,13α-endoperoxide (compound 1), was isolated from the stems of Microtoena insuavis (Hance) Prain ex Dunn, along with 4,4'-dihydroxytruxillic acid (compound 2), gallic acid (compound 3), ellagic acid (compound 4), 3-O-methylellagic acid 3'-O-α-rhamnopyranoside (compound 5), 3(''')-O-methylcrenatoside (compound 6), crenatoside (compound 7), apigenin (compound 8), succinic acid (compound 9), β-sitosterol (compound 10), and β-daucosterol (compound 11). The structures of these compounds were elucidated on the basis of spectral evidence.

  1. Norcembranoidal Diterpenes from the Cultured-Type Octocoral Sinularia numerosa

    Directory of Open Access Journals (Sweden)

    Wu-Fu Chen

    2015-02-01

    Full Text Available A known norcembranoidal diterpene, 5-episinuleptolide (1, along with a new analogue, 4α-hydroxy-5-episinuleptolide (2, were isolated from a cultured-type soft coral Sinularia numerosa. The structures of 1 and 2 were elucidated on the basis of spectroscopic methods and by comparison of the data with those of the related metabolites. Cytotoxicity of metabolites 1 and 2 against a panel of tumor cells is also described. Compound 2 exhibited moderate cytotoxicity toward CCRF-CEM cells with an IC50 value 4.21 μg/mL. Preliminary SAR (structure activity relationship information was obtained from these two compounds.

  2. Synthesis and sensory evaluation of ent-kaurane diterpene glycosides.

    Science.gov (United States)

    Prakash, Indra; Campbell, Mary; San Miguel, Rafael Ignacio; Chaturvedula, Venkata Sai Prakash

    2012-07-26

    Catalytic hydrogenation of the three ent-kaurane diterpene glycosides isolated from Stevia rebaudiana, namely rubusoside, stevioside, and rebaudioside-A has been carried out using Pd(OH)₂ and their corresponding dihydro derivatives have been isolated as the products. Synthesis of reduced steviol glycosides was performed using straightforward chemistry and their structures were characterized on the basis of 1D and 2D NMR spectral data and chemical studies. Also, we report herewith the sensory evaluation of all the reduced compounds against their corresponding original steviol glycosides and sucrose for the sweetness property of these molecules.

  3. Discovery of a Novel Diterpene in Brown Propolis from the State of Parana, Brazil.

    Science.gov (United States)

    Tazawa, Shigemi; Arai, Yasuko; Hotta, Sho; Mitsui, Taichi; Nozaki, Hiroshi; Ichihara, Kenji

    2016-02-01

    Propolis is a resinous substance collected by honeybees from certain plant sources. The components of propolis depend on the vegetation of the area in which apiculture is practiced. In Brazil, there are several types of propolis including 'green,' 'red' and 'brown'. Brazilian brown propolis from the state of Parana characteristically includes diterpenes, and we discovered a novel clerodane diterpene, rel-(5S,6S,8R,9R,10S,18R,19S)-18,19-epoxy-2-oxocleroda-3,12(E),14- triene-6,18,19-triol 18,19-diacetate 6-benzoate (3) and five known diterpenes (1, 2, 4, 5 and 6). The chemical structure of the novel diterpene 3 was determined using 1D- and 2D-NMR spectroscopic analyses. Furthermore, the activities of the isolated diterpenes on growth inhibition of several human cancer cell lines (LNCaP, MCF-7, DLD-1 and A549) were evaluated in vitro; diterpene 3 exhibited a potent inhibition of cell growth, and its activity was approximately 15 times higher than that of the other diterpenes.

  4. Sphaerococcenol A: a new rearranged bromo-diterpene from the red alga Sphaerococcus coronopifolius

    Energy Technology Data Exchange (ETDEWEB)

    Fenical, W. (Scripps Inst. of Oceanography, La Jolla, CA); Finer, J.; Clardy, J.

    1976-01-01

    Biosynthesis involving bromination mechanisms is now recognized as a characteristic of at least nine families of the red seaweeds (Rhodophyta). While bromine-containing diterpenes are not common among these algae, examples of two ring systems, concinndiol and iriediol, are elaborated only by Laurencia species. The structure of a new bromo-diterpene obtained from extracts of the red seaweed Sphaerococcus coronopifolius is reported. This metabolite appears to result from a methyl migration of a tail to tail diterpene precursor or from more substantial molecular rearrangements.

  5. Expanding the Landscape of Diterpene Structural Diversity through Stereochemically Controlled Combinatorial Biosynthesis.

    Science.gov (United States)

    Andersen-Ranberg, Johan; Kongstad, Kenneth Thermann; Nielsen, Morten Thrane; Jensen, Niels Bjerg; Pateraki, Irini; Bach, Søren Spanner; Hamberger, Britta; Zerbe, Philipp; Staerk, Dan; Bohlmann, Jörg; Møller, Birger Lindberg; Hamberger, Björn

    2016-02-01

    Plant-derived diterpenoids serve as important pharmaceuticals, food additives, and fragrances, yet their low natural abundance and high structural complexity limits their broader industrial utilization. By mimicking the modularity of diterpene biosynthesis in plants, we constructed 51 functional combinations of class I and II diterpene synthases, 41 of which are "new-to-nature". Stereoselective biosynthesis of over 50 diterpene skeletons was demonstrated, including natural variants and novel enantiomeric or diastereomeric counterparts. Scalable biotechnological production for four industrially relevant targets was accomplished in engineered strains of Saccharomyces cerevisiae.

  6. Minor diterpene glycosides from the leaves of Stevia rebaudiana.

    Science.gov (United States)

    Ibrahim, Mohamed A; Rodenburg, Douglas L; Alves, Kamilla; Fronczek, Frank R; McChesney, James D; Wu, Chongming; Nettles, Brian J; Venkataraman, Sylesh K; Jaksch, Frank

    2014-05-23

    Two new diterpene glycosides in addition to five known glycosides have been isolated from a commercial extract of the leaves of Stevia rebaudiana. Compound 1 (rebaudioside KA) was shown to be 13-[(O-β-d-glucopyranosyl)oxy]ent-kaur-16-en-19-oic acid 2-O-β-d-glucopyranosyl-β-d-glucopyranosyl ester and compound 2, 12-α-[(2-O-β-d-glucopyranosyl-β-d-glucopyranosyl)oxy]ent-kaur-16-en-19-oic acid β-d-glucopyranosyl ester. Five additional known compounds were identified, rebaudioside E, rebaudioside M, rebaudioside N, rebaudioside O, and stevioside, respectively. Enzymatic hydrolysis of stevioside afforded the known ent-kaurane aglycone 13-hydroxy-ent-kaur-16-en-19-oic acid (steviol) (3). The isolated metabolite 1 possesses the ent-kaurane aglycone steviol (3), while compound 2 represents the first example of the isomeric diterpene 12-α-hydroxy-ent-kaur-16-en-19-oic acid existing as a glycoside in S. rebaudiana. The structures of the isolated metabolites 1 and 2 were determined based on comprehensive 1D- and 2D-NMR (COSY, HSQC, and HMBC) studies. A high-quality crystal of compound 3 has formed, which allowed the acquisition of X-ray diffraction data that confirmed its structure. The structural similarities between the new metabolites and the commercially available stevioside sweeteners suggest the newly isolated metabolites should be examined for their organoleptic properties. Accordingly rebaudiosides E, M, N, O, and KA have been isolated in greater than gram quantities.

  7. Phytophthora austrocedri Elicitates Changes in Diterpene Profile of Austrocedrus chilensis.

    Science.gov (United States)

    Olate, Verónica Rachel; Vélez, María Laura; Greslebin, Alina; Schmeda-Hirschmann, Guillermo

    2015-08-18

    The populations of the Andean Cupressaceae Austrocedrus chilensis have been severely affected by a disease caused by the phytopathogenic fungus Phytophthora austrocedri. A study was undertaken to disclose changes in the resin composition of P. austrocedri-infected individuals, including naturally infected and artificially inoculated trees, compared with healthy A. chilensis trees. GC-MS and (1)H-NMR studies showed a clear differentiation among healthy and infected resins, with the diterpene isopimara-8(9),15-dien-19-ol as a relevant constituent in resins from infected trees. The effect of resin fractions from P. austrocedri infected trees on the pathogen was assessed by measuring the mycelial growth in agar plates. The most active fractions from resin obtained from infected trees inhibited fungal growth by nearly 50% at 1 mg/dish (35.37 µg/cm(2)). The main constituent in the active fractions were 18-hydroxymanool and the aldehyde torulosal. Both compounds are oxidation products of manool and can be a chemical response of the tree to the pathogen or be formed from the pathogen as a biotransformation product of manool by microbial oxidation. While the diterpene profiles from A. chilensis tree resins can easily differentiate healthy and P. austrocedri infected individuals, the possible conversion of manool to the antifungal derivatives 4 and 6 by the microorganism remains to be established.

  8. Phytophthora austrocedri Elicitates Changes in Diterpene Profile of Austrocedrus chilensis

    Directory of Open Access Journals (Sweden)

    Verónica Rachel Olate

    2015-08-01

    Full Text Available The populations of the Andean Cupressaceae Austrocedrus chilensis have been severely affected by a disease caused by the phytopathogenic fungus Phytophthora austrocedri. A study was undertaken to disclose changes in the resin composition of P. austrocedri-infected individuals, including naturally infected and artificially inoculated trees, compared with healthy A. chilensis trees. GC-MS and 1H-NMR studies showed a clear differentiation among healthy and infected resins, with the diterpene isopimara-8(9,15-dien-19-ol as a relevant constituent in resins from infected trees. The effect of resin fractions from P. austrocedri infected trees on the pathogen was assessed by measuring the mycelial growth in agar plates. The most active fractions from resin obtained from infected trees inhibited fungal growth by nearly 50% at 1 mg/dish (35.37 µg/cm2. The main constituent in the active fractions were 18-hydroxymanool and the aldehyde torulosal. Both compounds are oxidation products of manool and can be a chemical response of the tree to the pathogen or be formed from the pathogen as a biotransformation product of manool by microbial oxidation. While the diterpene profiles from A. chilensis tree resins can easily differentiate healthy and P. austrocedri infected individuals, the possible conversion of manool to the antifungal derivatives 4 and 6 by the microorganism remains to be established.

  9. Six new cassane diterpenes from the twigs and leaves of Tara (Caesalpinia spinosa Kuntze).

    Science.gov (United States)

    He, Dongyang; Li, Yuanping; Tang, Hongbo; Ma, Ruijing; Li, Xiaonian; Wang, Liqin

    2015-09-01

    Six new cassane diterpenes, isoneocaesalpin H (1), caespinosin A (2), caespinosin B (3), a cassane diterpene with unique 6/6/7 carbon rings, and caespinosins C-E (4-6) were isolated from the twigs and leaves of Tara (Caesalpinia spinosa Kuntze). The absolute configuration of isoneocaesalpin H (1) was determined by single-crystal X-ray crystallographic analysis. Compound 3 represents a class of rare natural cassane diterpene bearing unique 6/6/7 carbon rings. Their structures were identified by 1D and 2D NMR spectral data. Cassane diterpenes were firstly reported from Tara. Compounds 1-5 were evaluated for their cytotoxicity on HL-60, SMMC-7721, A-549, MCF-7 and SW-480 human cancer cell lines, but they were inactive.

  10. Structure of corylifuran: a clerodane-type diterpene from Croton corylifolius Lam

    Energy Technology Data Exchange (ETDEWEB)

    Burke, B.A. (Univ. of the West Indies, Kingston, Jamaica); Chan, W.R.; Prince, E.C.; Manchand, P.S.; Eickman, N.; Clardy, J.

    1976-01-01

    The structure and stereochemistry of corylifuran, a clerodane-type diterpene from Croton corylifolius Lam, have been determined as 2 by chemical and spectroscopic data and X-ray crystallographic analysis.

  11. The Reduction of a Natural Diterpene Containing o, β-Unsaturated Keto Group

    Institute of Scientific and Technical Information of China (English)

    Qing ZHAO; Xin HONG; Cheng ZOU; Yue Mao SHEN; Xiao Jiang HAO

    2003-01-01

    Hedychenone (1), a diterpene containing α, β-unsaturated keto group, was reduced byaluminum-mercury alloy, and a dimerized product (2) was obtained as the major product. Thecoupling occurred at β position of the keto group.

  12. Two new acyclic diterpene-y-lactones from the leaves of Salix matsudana

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Two new acyclic diterpene-(-lactones named hanliuine III (1) and hanliuine IV (2) were isolated from leaves of Salix matsudana (Chinese name "hanliu"). Their structures were deduced from spectral data.

  13. Extreme promiscuity of a bacterial and a plant diterpene synthase enables combinatorial biosynthesis.

    Science.gov (United States)

    Jia, Meirong; Potter, Kevin C; Peters, Reuben J

    2016-09-01

    Diterpenes are widely distributed across many biological kingdoms, where they serve a diverse range of physiological functions, and some have significant industrial utility. Their biosynthesis involves class I diterpene synthases (DTSs), whose activity can be preceded by that of class II diterpene cyclases (DTCs). Here, a modular metabolic engineering system was used to examine the promiscuity of DTSs. Strikingly, both a bacterial and plant DTS were found to exhibit extreme promiscuity, reacting with all available precursors with orthogonal activity, producing an olefin or hydroxyl group, respectively. Such DTS promiscuity enables combinatorial biosynthesis, with remarkably high yields for these unoptimized non-native enzymatic combinations (up to 15mg/L). Indeed, it was possible to readily characterize the 13 unknown products. Notably, 16 of the observed diterpenes were previously inaccessible, and these results provide biosynthetic routes that are further expected to enable assembly of more extended pathways to produce additionally elaborated 'non-natural' diterpenoids.

  14. A NEW PHENOLIC GLYCOSIDE AND A NEW TRANS-CLERODANE DITERPENE FROM CONYZA BLINII

    Institute of Scientific and Technical Information of China (English)

    YAN-FANG SU; DEAN GUO; YA-JUN CUI; JIAN-SHENG LIU; JUN-HUA ZHENG

    2001-01-01

    A new phenolic glycoside, 4-propionyl-2,6-dimethoxyphenyl β-D-glucopyranoside (1) and a new trans-clerodane diterpene named 19-deacetylconyzalactone (2), were isolated from the aerial parts of Conyza blinii.

  15. Synthetic studies of neoclerodane diterpenes from Salvia divinorum: preparation and opioid receptor activity of salvinicin analogues.

    Science.gov (United States)

    Simpson, Denise S; Katavic, Peter L; Lozama, Anthony; Harding, Wayne W; Parrish, Damon; Deschamps, Jeffrey R; Dersch, Christina M; Partilla, John S; Rothman, Richard B; Navarro, Hernan; Prisinzano, Thomas E

    2007-07-26

    Further modification of salvinorin A (1a), the major active component of Salvia divinorum, has resulted in the synthesis of novel neoclerodane diterpenes with opioid receptor affinity and activity. We report in this study that oxadiazole 11a and salvidivin A (12a), a photooxygenation product of 1a, have been identified as the first neoclerodane diterpenes with kappa antagonist activity. This indicates that additional structural modifications of 1a may lead to analogues with higher potency and utility as drug abuse medications.

  16. Structure and mechanism of the diterpene cyclase ent-copalyl diphosphate synthase

    Energy Technology Data Exchange (ETDEWEB)

    Köksal, Mustafa; Hu, Huayou; Coates, Robert M.; Peters, Reuben J.; Christianson, David W. (UIUC); (Iowa State); (Penn)

    2011-09-20

    The structure of ent-copalyl diphosphate synthase reveals three {alpha}-helical domains ({alpha}, {beta} and {gamma}), as also observed in the related diterpene cyclase taxadiene synthase. However, active sites are located at the interface of the {beta}{gamma} domains in ent-copalyl diphosphate synthase but exclusively in the {alpha} domain of taxadiene synthase. Modular domain architecture in plant diterpene cyclases enables the evolution of alternative active sites and chemical strategies for catalyzing isoprenoid cyclization reactions.

  17. Structure and Mechanism of the Diterpene Cyclase ent-Copalyl Diphosphate Synthase

    Science.gov (United States)

    Köksal, Mustafa; Hu, Huayou; Coates, Robert M.; Peters, Reuben J.; Christianson, David W.

    2011-01-01

    The structure of ent-copalyl diphosphate synthase (CPS) reveals three α-helical domains (α, β, γ), as also observed in the related diterpene cyclase taxadiene synthase. However, active sites are located at the interface of the βγ domains in CPS but exclusively in the α domain of taxadiene synthase. Modular domain architecture in plant diterpene cyclases enables the evolution of alternative active sites and chemical strategies for catalyzing isoprenoid cyclization reactions. PMID:21602811

  18. Kaurane diterpenes protect against apoptosis and inhibition of phagocytosis in activated macrophages.

    Science.gov (United States)

    de las Heras, B; Hortelano, S; Girón, N; Bermejo, P; Rodríguez, B; Boscá, L

    2007-09-01

    The kaurane diterpenes foliol and linearol are inhibitors of the activation of nuclear factor kappaB, a transcription factor involved in the inflammatory response. Effects of these diterpenes on apoptosis and phagocytosis have been analysed in cultured peritoneal macrophages and in the mouse macrophage cell line, RAW 264.7. Macrophages were maintained in culture and activated with pro-inflammatory stimuli in the absence or presence of diterpenes. Apoptosis and the phagocytosis in these cells under these conditions were determined. Incubation of macrophages with a mixture of bacterial lipopolysaccharide (LPS)/interferon-gamma (IFN-gamma) induced apoptosis through a NO-dependent pathway, an effect significantly inhibited by foliol and linearol in the low muM range, without cytotoxic effects. Apoptosis in macrophages induced by NO donors was also inhibited. The diterpenes prevented apoptosis through a mechanism compatible with the inhibition of caspase-3 activation, release of cytochrome c to the cytosol and p53 overexpression, as well as an alteration in the levels of proteins of the Bcl-2 family, in particular, the levels of Bax. Cleavage of poly(ADP-ribose) polymerase, a well-established caspase substrate, was reduced by these diterpenes. Treatment of cells with foliol and linearol decreased phagocytosis of zymosan bioparticles by RAW 264.7 cells and to a greater extent by peritoneal macrophages. Both diterpenes protected macrophages from apoptosis and inhibited phagocytosis, resulting in a paradoxical control of macrophage function, as viability was prolonged but inflammatory and phagocytic functions were impaired.

  19. Molecular Signaling Pathways Behind the Biological Effects of Salvia Species Diterpenes in Neuropharmacology and Cardiology.

    Science.gov (United States)

    Akaberi, M; Iranshahi, M; Mehri, S

    2016-06-01

    The genus Salvia, from the Lamiaceae family, has diverse biological properties that are primarily attributable to their diterpene contents. There is no comprehensive review on the molecular signaling pathways of these active components. In this review, we investigated the molecular targets of bioactive Salvia diterpenes responsible for the treatment of nervous and cardiovascular diseases. The effects on different pathways, including apoptosis signaling, oxidative stress phenomena, the accumulation of amyloid beta plaques, and tau phosphorylation, have all been considered to be mechanisms of the anti-Alzheimer properties of Salvia diterpenes. Additionally, effects on the benzodiazepine and kappa opioid receptors and neuroprotective effects are noted as neuropharmacological properties of Salvia diterpenes, including tanshinone IIA, salvinorin A, cryptotanshinone, and miltirone. Tanshinone IIA, as the primary diterpene of Salvia miltiorrhiza, has beneficial activities in heart diseases because of its ability to scavenge free radicals and its effects on transcription factors, such as nuclear transcription factor-kappa B (NF-κB) and the mitogen-activated protein kinases (MAPKs). Additionally, tanshinone IIA has also been proposed to have cardioprotective properties including antiarrhythmic activities and effects on myocardial infarction. With respect to the potential therapeutic effects of Salvia diterpenes, comprehensive clinical trials are warranted to evaluate these valuable molecules as lead compounds. Copyright © 2016 John Wiley & Sons, Ltd.

  20. Chemosystematic investigations of irregular diterpenes in Anisotome and related New Zealand Apiaceae.

    Science.gov (United States)

    Zidorn, Christian; Sturm, Sonja; Dawson, John W; van Klink, John W; Stuppner, Hermann; Perry, Nigel B

    2002-02-01

    A chemosystematic HPLC-UV and HPLC-MS investigation of New Zealand members of the Apiaceae was performed. Diterpenes were identified and quantified in methanolic extracts from subaerial parts of 28 taxa and 54 samples of Aciphylla, Anisotome, Apium, Gingidia, Lignocarpa, Oreomyrrhis, and Scandia. Six diterpenes (1-2, 4-7) and four polyacetylenes (8-11) were identified. The known compounds were the diterpenes anisotomenoic acid 1, anisotomene-1-ol 2, 16-acetoxyanisotomenoic acid 4 and anisotomene-1,12-diol 5; and the polyacetylenes falcarinol 8, falcarindiol 9, (+)-9(Z),17-octadecadiene-12,14-diyne-1,11,16-triol 10, and (+)-9(Z),17-octadecadiene-12,14-diyne-1,11,16-triol 1-acetate 11. New irregular diterpenes 13,14-dihydroanisotom-12E-ene-1,14-diol 6 and 14-methoxy-13,14-dihydroanisotom-12E-ene-1-ol 7 were isolated from A. haastii. Isomers of the new semi-synthetic diterpene 16-hydroxyanisotomenoic acid 3 were detected in extracts of Anisitone flexuosa. Structure elucidation was performed by HR mass spectrometry and 1D and 2D NMR spectroscopy. In crude extracts, compounds were identified by their HPLC retention times and their on-line HPLC-UV and MS spectra. Anisotomene diterpenes occurred in eight out of 16 species of the genus Anisotome, but were not detected in any of the other genera. In contrast, polyacetylenes were present in all the genera investigated.

  1. Variability of some diterpene esters in coffee beverages as influenced by brewing procedures.

    Science.gov (United States)

    Moeenfard, Marzieh; Erny, Guillaume L; Alves, Arminda

    2016-11-01

    Several coffee brews, including classical and commercial beverages, were analyzed for their diterpene esters content (cafestol and kahweol linoleate, oleate, palmitate and stearate) by high performance liquid chromatography with diode array detector (HPLC-DAD) combined with spectral deconvolution. Due to the coelution of cafestol and kahweol esters at 225 nm, HPLC-DAD did not give accurate quantification of cafestol esters. Accordingly, spectral deconvolution was used to deconvolve the co-migrating profiles. Total cafestol and kahweol esters content of classical coffee brews ranged from 5-232 to 2-1016 mg/L, respectively. Commercial blends contained 1-54 mg/L of total cafestol esters and 2-403 mg/L of total kahweol esters. Boiled coffee had the highest diterpene esters content, while filtered and instant brews showed the lowest concentrations. However, individual diterpene esters content was not affected by brewing procedure as in terms of kahweol esters, kahweol palmitate was the main compound in all samples, followed by kahweol linoleate, oleate and stearate. Higher amounts of cafestol palmitate and stearate were also observed compared to cafestol linoleate and cafestol oleate. The ratio of diterpene esters esterified with unsaturated fatty acids to total diterpene esters was considered as measure of their unsaturation in analyzed samples which varied from 47 to 52%. Providing new information regarding the diterpene esters content and their distribution in coffee brews will allow a better use of coffee as a functional beverage.

  2. Pharmacological studies on coleonol, a hypotensive diterpene from Coleus forskohlii.

    Science.gov (United States)

    Dubey, M P; Srimal, R C; Nityanand, S; Dhawan, B N

    1981-01-01

    Coleus spp. have been used in Aurvedic medicine for heart diseases, spasmodic pain, painful micturition and convulsions. The pharmacological properties of coleonol, a diterpene, isolated from Coleus forskohlii were investigated. Its predominant effect is to lower the blood pressure of anaesthetised cat and rat as well as of the spontaneously hypertensive rat due to relaxation of the vascular smooth muscle. In small doses it has a positive inotropic effect on isolated rabbit heart as well as on cat heart in vivo. Coleonol also exhibits nonspecific spasmolytic activity on smooth muscle of the gastrointestinal tract in various species but not on bronchial musculature of guinea pig. Large doses of coleonol have a depressant action on the central nervous system. These results provide the rationale for the use of this plant in Aurvedic medicine.

  3. Identification of some novel tetracyclic diterpene hydrocarbons in petroleum

    Energy Technology Data Exchange (ETDEWEB)

    Petrov, A.A.; Pehk, T.Y.; Vorobieva, N.S.; Zemskova, Z.K. (Institute of Geology and Exploitation of Combustible Minerals, Moscow (USSR))

    1988-01-01

    A new group of tetracyclic diterpene hydrocarbons of molecular formula C{sub 19}H{sub 32} has been found in the Jurassic oils and condensates of the Central Kara-Kum (Turkmenia, U.S.S.R.). The structure of the hydrocarbons has been determined by gas chromatography-mass spectrometry and {sup 1}H and {sup 13}C NMR. Of the compounds identified 4,8-dimethyl-13-isopropyltetracyclo(6.6.0.0{sup 1,11}O{sup 3,7})- and 5,14-dimethyl-10-isopropyltetracyclo = (6.4.1.1.{sup 1,9}O{sup 4,13})tetradecanes are present in the highest concentrations. Some ideas are put forward about the source and the reactions involved in the formation of the hydrocarbons under natural conditions by the enzymic C{sub 5} cyclization of aliphatic isoprenoids.

  4. Vibrational spectra and DFT calculations of sonderianin diterpene

    Science.gov (United States)

    Oliveira, I. M. M.; Santos, H. S.; Sena, D. M.; Cruz, B. G.; Teixeira, A. M. R.; Freire, P. T. C.; Braz-Filho, R.; Sousa, J. W.; Albuquerque, M. R. J. R.; Bandeira, P. N.; Bernardino, A. C. S. S.; Gusmão, G. O. M.; Bento, R. R. F.

    2015-11-01

    In the present study, the natural product sonderianin diterpene (C21H26O4), a diterpenoid isolated from Croton blanchetianus, with potential application in the drug industry, was characterized by nuclear magnetic resonance, infrared and Raman spectroscopy. Vibrational spectra were supported by Density Functional Theory calculations. Infrared and Raman spectra of sonderianin were recorded at ambient temperature in the regions from 400 cm-1 to 3600 cm-1 and from 40 cm-1 to 3500 cm-1, respectively. DFT calculations with the hybrid functional B3LYP and the basis set 6-31 G(d,p) were performed with the purpose of obtaining information on the structural and vibrational properties of this organic compound. A comparison with experimental spectra allowed us to assign all of the normal modes of the crystal. The assignment of the normal modes was carried out by means of potential energy distribution.

  5. Diterpene and other constituents from Stemodia maritima (Scrophulariaceae)

    Energy Technology Data Exchange (ETDEWEB)

    Rodrigues, Francisco E.A.; Oliveira, Maria da Conceicao F. de; Vasconcelos, Jackson N.; Mafezoli, Jair; Arriaga, Angela M.C., E-mail: angelamcarriaga@yahoo.com.b [Universidade Federal do Ceara (DQOI/UFC), Fortaleza, CE (Brazil). Dept. de Quimica Organica e Inorganica. Curso de Pos-Graducao em Quimica; Lima, Jefferson Q. [Instituto Federal do Ceara, Juazeiro do Norte, CE (Brazil). Curso de Engenharia Ambiental; Santiago, Gilvandete M.P. [Universidade Federal do Ceara (DQOI/UFC), Fortaleza, CE (Brazil). Dept. de Farmacia; Braz-Filho, Raimundo [Universidade Estadual do Norte Fluminense Darcy Ribeiro (CCT/UENF), Campos dos Goytacazes, RJ (Brazil). Centro de Ciencias e Tecnologias

    2010-07-01

    A new diterpene, (5S{sup *},8S{sup *},9R{sup *},10S{sup *})-11{beta},12{beta}-epoxy-9{alpha}-hydroxy-19(4 -> 3)abeo-abieta-3,13-diene-19,18-olide, together with the known compounds stemodin, D-mannitol, betulinic acid, a mixture of 3{beta}-O-{beta}-D-glucopyranosyl-{beta}-sitosterol and 3{beta}-O-{beta}-D-glucopyranosylstigmasterol and 5,7,4'-trihydroxy-3,8,3'-trimethoxyflavone were isolated from the leaves and stems of Stemodia maritima. Structural elucidation of all compounds was based on interpretation of spectral data, mainly NMR (1D and 2D) and MS, including comparison with values described in the literature. (author)

  6. Selective activation of liver X receptors by acanthoic acid-related diterpenes.

    Science.gov (United States)

    Traves, Paqui G; Hortelano, Sonsoles; Zeini, Miriam; Chao, Ta-Hsiang; Lam, Thanh; Neuteboom, Saskia T; Theodorakis, Emmanuel A; Palladino, Michael A; Castrillo, Antonio; Bosca, Lisardo

    2007-06-01

    Terpenoids constitute a large family of natural steroids that are widely distributed in plants and insects. We investigated the effects of a series of diterpenes structurally related to acanthoic acid in macrophage functions. We found that diterpenes with different substitutions at the C4 position in ring A are potent activators of liver X receptors (LXRalpha and LXRbeta) in both macrophage cell lines from human and mouse origin and primary murine macrophages. Activation of LXR by these diterpenes was evaluated in transient transfection assays and gene expression analysis of known LXR-target genes, including the cholesterol transporters ABCA1 and ABCG1, the sterol regulatory element-binding protein 1c, and the apoptosis inhibitor of macrophages (Spalpha). Moreover, active diterpenes greatly stimulated cholesterol efflux from macrophages. It is interesting that these diterpenes antagonize inflammatory gene expression mainly through LXR-dependent mechanisms, indicating that these compounds can activate both LXR activation and repression functions. Stimulation of macrophages with acanthoic acid diterpenes induced LXR-target gene expression and cholesterol efflux to similar levels observed with synthetic agonists 3-[3-[N-(2-chloro-3-trifluoromethylbenzyl)-(2,2-diphenylethyl)-amino]propyloxy]phenylacetic acid hydrochloride (GW3965) and N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)-ethyl]phenyl]-benzenesulfonamide [T1317 (T0901317)]. These effects observed in gene expression were deficient in macrophages lacking both LXR isoforms (LXRalpha,beta(-/-)). These results show the ability of certain acanthoic acid diterpenes to activate efficiently both LXRs and suggest that these compounds can exert beneficial effects from a cardiovascular standpoint through LXR-dependent mechanisms.

  7. Effects of diterpene acids on components of a conifer bark beetle-fungal interaction : tolerance by Ips pini and sensitivity by its associate Ophiostoma ips

    Science.gov (United States)

    Brian J. Kopper; Barbara L. Illman; Philip J. Kersten; Kier D. Klepzig; Kenneth F. Raffa

    2005-01-01

    Conifer resin and phloem tissue contain several phytochemical groups, composed primarily of monoterpenes, diterpene acids, and stilbene phenolics. The effects of monoterpenes and phenolics on stem-colonizing bark beetles and their associated microorganisms have been studied to some extent, but the roles of diterpene acids are largely unknown. Diterpene acids are known...

  8. Effects of diterpene acids on components of a conifer bark beetle–fungal interaction: tolerance by Ips pini and sensitivity by its associate Ophiostoma ips

    Science.gov (United States)

    Brian J. Kopper; Barbara L. Illman; Philip J. Kersten; Kier D. Klepzig; Kenneth F. Raffa

    2005-01-01

    Conifer resin and phloem tissue contain several phytochemical groups,composed primarily of monoterpenes,diterpene acids, and stilbene phenolics. The effects of monoterpenes and phenolics on stem-colonizing bark beetles and their associated microorganisms have been studied to some extent, but the roles of diterpene acids are largely unknown. Diterpene acids are known to...

  9. Antiplatelet and Anticoagulant Effects of Diterpenes Isolated from the Marine Alga, Dictyota menstrualis

    Directory of Open Access Journals (Sweden)

    Laura de Andrade Moura

    2014-04-01

    Full Text Available Cardiovascular diseases represent a major cause of disability and death worldwide. Therapeutics are available, but they often have unsatisfactory results and may produce side effects. Alternative treatments based on the use of natural products have been extensively investigated, because of their low toxicity and side effects. Marine organisms are prime candidates for such products, as they are sources of numerous and complex substances with ecological and pharmacological effects. In this work, we investigated, through in vitro experiments, the effects of three diterpenes (pachydictyol A, isopachydictyol A and dichotomanol from the Brazilian marine alga, Dictyota menstrualis, on platelet aggregation and plasma coagulation. Results showed that dichotomanol inhibited ADP- or collagen-induced aggregation of platelet-rich plasma (PRP, but failed to inhibit washed platelets (WP. In contrast, pachydictyol A and isopachydictyol A failed to inhibit the aggregation of PRP, but inhibited WP aggregation induced by collagen or thrombin. These diterpenes also inhibited coagulation analyzed by the prothrombin time and activated partial thromboplastin time and on commercial fibrinogen. Moreover, diterpenes inhibited the catalytic activity of thrombin. Theoretical studies using the Osiris Property Explorer software showed that diterpenes have low theoretical toxicity profiles and a drug-score similar to commercial anticoagulant drugs. In conclusion, these diterpenes are promising candidates for use in anticoagulant therapy, and this study also highlights the biotechnological potential of oceans and the importance of bioprospecting to develop medicines.

  10. 二萜合酶的研究进展%Research advances of diterpene synthase

    Institute of Scientific and Technical Information of China (English)

    何云飞; 高伟; 刘塔斯; 李文渊; 黄璐琦

    2011-01-01

    Diterpenes, an important class of natural compounds, are widely distributed in nature. As the valuable diterpenoids continue to be found, diterpene synthase in the course of diterpene synthesis get as much attention as possible. The multiformity of end-product-diterpenoids were also due to the diversity of diterpene synthase. This paper focuses on the advances in recent biosynthesis pathway of diterpene and types, cloning, catalytic mechanism, synthetic biology application.%植物二萜类化合物广泛存在于自然界中,是一类重要的天然化合物.随着一些具有重要经济价值的二萜不断被发现,其生物合成中的二萜合酶也倍受关注.二萜合酶催化活性的多样性决定了终产物(二萜化合物)的丰富性.本文重点探讨了近年来二萜生物合成途径和二萜合酶的类型、克隆、催化机制及其合成生物学应用等研究进展.

  11. Indole-Diterpene Biosynthetic Capability of Epichloë Endophytes as Predicted by ltm Gene Analysis▿

    Science.gov (United States)

    Young, Carolyn A.; Tapper, Brian A.; May, Kimberley; Moon, Christina D.; Schardl, Christopher L.; Scott, Barry

    2009-01-01

    Bioprotective alkaloids produced by Epichloë and closely related asexual Neotyphodium fungal endophytes protect their grass hosts from insect and mammalian herbivory. One class of these compounds, known for antimammalian toxicity, is the indole-diterpenes. The LTM locus of Neotyphodium lolii (Lp19) and Epichloë festuce (Fl1), required for the biosynthesis of the indole-diterpene lolitrem, consists of 10 ltm genes. We have used PCR and Southern analysis to screen a broad taxonomic range of 44 endophyte isolates to determine why indole-diterpenes are present in so few endophyte-grass associations in comparison to that of the other bioprotective alkaloids, which are more widespread among the endophtyes. All 10 ltm genes were present in only three epichloë endophytes. A predominance of the asexual Neotyphodium spp. examined contained 8 of the 10 ltm genes, with only one N. lolii containing the entire LTM locus and the ability to produce lolitrems. Liquid chromatography-tandem mass spectrometry profiles of indole-diterpenes from a subset of endophyte-infected perennial ryegrass showed that endophytes that contained functional genes present in ltm clusters 1 and 2 were capable of producing simple indole-diterpenes such as paspaline, 13-desoxypaxilline, and terpendoles, compounds predicted to be precursors of lolitrem B. Analysis of toxin biosynthesis genes by PCR now enables a diagnostic method to screen endophytes for both beneficial and detrimental alkaloids and can be used as a resource for screening isolates required for forage improvement. PMID:19181837

  12. Additional Minor Diterpene Glycosides from Stevia rebaudiana Bertoni

    Directory of Open Access Journals (Sweden)

    Indra Prakash

    2013-10-01

    Full Text Available Two additional novel minor diterpene glycosides were isolated from the commercial extract of the leaves of Stevia rebaudiana Bertoni. The structures of the new compounds were identified as 13-{β-D-glucopyranosyl-(1→2-O-[β-D-glucopyranosyl-(1→3-β-D-glucopyranosyl-oxy} ent-kaur-16-en-19-oic acid {β-D-xylopyranosyl-(1→2-O-[β-D-glucopyranosyl-(1→3]-O-β-D-glucupyranosyl-ester} (1, and 13-{β-D-6-deoxy-glucopyranosyl-(1→2-O-[β-D-glucopyranosyl-(1→3-β-D-glucopyranosyl-oxy} ent-kaur-16-en-19-oic acid {β-D-glucopyranosyl-(1→2-O-[β-D-glucopyranosyl-(1→3-β-D-gluco-pyranosyl-ester} (2, on the basis of extensive 1D (1H- and 13C- 2D NMR (COSY, HSQC and HMBC and MS spectroscopic data as well as chemical studies.

  13. Three New Clerodane Diterpenes from Polyalthia longifolia var. pendula

    Directory of Open Access Journals (Sweden)

    Tung-Ho Wu

    2014-02-01

    Full Text Available Three new clerodane diterpenes, (4→2-abeo-cleroda-2,13E-dien-2,14-dioic acid (1, (4→2-abeo-2,13-diformyl-cleroda-2,13E-dien-14-oic acid (2, and 16(R&S- methoxycleroda-4(18,13-dien-15,16-olide (3, were isolated from the unripe fruit of Polyalthia longifolia var. pendula (Annonaceae together with five known compounds (4–8. The structures of all isolates were determined by spectroscopic analysis. The anti-inflammatory activity of the isolates was evaluated by testing their inhibitory effect on NO production in LPS-stimulated RAW 264.7 macrophages. Among the isolated compounds, 16-hydroxycleroda-3,13-dien-15,16-olide (6 and 16-oxocleroda-3,13-dien-15-oic acid (7 showed promising NO inhibitory activity at 10 µg/mL, with 81.1% and 86.3%, inhibition, respectively.

  14. Three New Cytotoxic ent-Kaurane Diterpenes from Isodon excisoides.

    Science.gov (United States)

    Dai, Li-Ping; Li, Chun; Yang, Han-Ze; Lu, Yan-Qing; Yu, Hong-Yan; Gao, Hui-Min; Wang, Zhi-Min

    2015-09-22

    Three types of ent-kaurane diterpenoids were isolated from the aerial parts of Isodon excisoides, including three new diterpenoids, 1α,7α,14β-trihydroxy-20-acetoxy-ent-kaur-15-one (1); 1α,7α,14β,18-tetrahydroxy-20-acetoxy-ent-kaur-15-one (2); and 1α-acetoxy-14β-hydroxy-7α,20-epoxy-ent-kaur-16-en-15-one (3); together with six known diterpenes henryin (4); kamebanin (5); reniformin C (6); kamebacetal A (7); kamebacetal B (8); and oridonin (9). The structures of the isolated compounds were elucidated by means of nuclear magnetic resonance spectroscopy and high-resolution mass spectrometry in conjunction with published data for their analogs, as well as their fragmentation patterns. Compounds 5 and 9 were isolated from Isodon excisoides for the first time. To explore the structure-activity relationships of the isolated compounds, they were tested for their cytotoxic effects against five human cancer cell lines: HCT-116, HepG2, A2780, NCI-H1650, and BGC-823. Most of the isolated compounds showed certain cytotoxic activity against the five cancer cell lines with IC50 values ranging from 1.09-8.53 µM. Among the tested compounds, compound 4 exhibited the strongest cytotoxic activity in the tested cell lines, with IC50 values ranging from 1.31-2.07 µM. Compounds 1, 6, and 7 exhibited selective cytotoxic activity.

  15. Carnosol: A Phenolic Diterpene With Cancer Chemopreventive Potential

    Science.gov (United States)

    Chun, Kyung-Soo; Kundu, Juthika; Chae, In Gyeong; Kundu, Joydeb Kumar

    2014-01-01

    Cancer is an unbeaten health challenge for the humankind. After striving for decades to find a cancer cure, attention has now been shifted to reduce the morbidity and mortality from cancer by halting the course of tumor development. Numerous bioactive phytochemicals, especially those present in edible and non-edible plant species, have been reported to reduce the risk of many cancers. Multiple lines of evidence suggest that carnosol, a phenolic diterpene present in rosemary (Rosmarinus officinalis L.), holds the promise of preventing certain types of cancer. A remarkable progress has been made in delineating the biochemical mechanisms underlying the chemopreventive effects of carnosol. Results from in vitro cell culture studies as well as animal model experiments have revealed that carnosol inhibits experimentally induced carcinogenesis and exhibits potent anti-oxidative, anti-inflammatory, antiproliferative and apoptosis inducing properties. Moreover, carnosol enhances the sensitivity of chemoresistant cancer cells to chemotherapeutic agents. The purpose of this review is to shed light on the detailed mechanistic aspects of cancer chemoprevention with carnosol. PMID:25337578

  16. Three New Cytotoxic ent-Kaurane Diterpenes from Isodon excisoides

    Directory of Open Access Journals (Sweden)

    Li-Ping Dai

    2015-09-01

    Full Text Available Three types of ent-kaurane diterpenoids were isolated from the aerial parts of Isodon excisoides, including three new diterpenoids, 1α,7α,14β-trihydroxy-20-acetoxy-ent-kaur-15-one (1; 1α,7α,14β,18-tetrahydroxy-20-acetoxy-ent-kaur-15-one (2; and 1α-acetoxy-14β-hydroxy-7α,20-epoxy-ent-kaur-16-en-15-one (3; together with six known diterpenes henryin (4; kamebanin (5; reniformin C (6; kamebacetal A (7; kamebacetal B (8; and oridonin (9. The structures of the isolated compounds were elucidated by means of nuclear magnetic resonance spectroscopy and high-resolution mass spectrometry in conjunction with published data for their analogs, as well as their fragmentation patterns. Compounds 5 and 9 were isolated from Isodon excisoides for the first time. To explore the structure-activity relationships of the isolated compounds, they were tested for their cytotoxic effects against five human cancer cell lines: HCT-116, HepG2, A2780, NCI-H1650, and BGC-823. Most of the isolated compounds showed certain cytotoxic activity against the five cancer cell lines with IC50 values ranging from 1.09–8.53 µM. Among the tested compounds, compound 4 exhibited the strongest cytotoxic activity in the tested cell lines, with IC50 values ranging from 1.31–2.07 µM. Compounds 1, 6, and 7 exhibited selective cytotoxic activity.

  17. Identification of a new diterpene biosynthetic gene cluster that produces O-methylkolavelool in Herpetosiphon aurantiacus.

    Science.gov (United States)

    Nakano, Chiaki; Oshima, Misaki; Kurashima, Nodoka; Hoshino, Tsutomu

    2015-03-23

    Diterpenoids are usually found in plants and fungi, but are rare in bacteria. We have previously reported new diterpenes, named tuberculosinol and isotuberculosinol, which are generated from the Mycobacterium tuberculosis gene products Rv3377c and Rv3378c. No homologous gene was found at that time, but we recently found highly homologous proteins in the Herpetosiphon aurantiacus ATCC 23779 genome. Haur_2145 was a class II diterpene cyclase responsible for the conversion of geranylgeranyl diphosphate into kolavenyl diphosphate. Haur_2146, homologous to Rv3378c, synthesized (+)-kolavelool through the nucleophilic addition of a water molecule to the incipient cation formed after the diphosphate moiety was released. Haur_2147 afforded (+)-O-methylkolavelool from (+)-kolavelool, so this enzyme was an O-methyltransferase. This new diterpene was indeed detected in H. aurantiacus cells. This is the first report of the identification of a (+)-O-methylkolavelool biosynthetic gene cluster.

  18. Additional diterpenes from Physcomitrella patens synthesized by copalyl diphosphate/kaurene synthase (PpCPS/KS).

    Science.gov (United States)

    Zhan, Xin; Bach, Søren Spanner; Hansen, Nikolaj Lervad; Lunde, Christina; Simonsen, Henrik Toft

    2015-11-01

    The bifunctional diterpene synthase, copalyl diphosphate/kaurene synthase from the moss Physcomitrella patens (PpCPS/KS), catalyses the formation of at least four diterpenes, including ent-beyerene, ent-sandaracopimaradiene, ent-kaur-16-ene, and 16-hydroxy-ent-kaurene. The enzymatic activity has been confirmed through generation of a targeted PpCPS/KS knock-out mutant in P. patens via homologous recombination, through transient expression of PpCPS/KS in Nicotiana benthamiana, and expression of PpCPS/KS in E. coli. GC-MS analysis of the knock-out mutant shows that it lacks the diterpenoids, supporting that all are products of PpCPS/KS as observed in N. benthamiana and E. coli. These results provide additional knowledge of the mechanism of this bifunctional diterpene synthase, and are in line with proposed reaction mechanisms in kaurene biosynthesis.

  19. Broad tuning of the human bitter taste receptor hTAS2R46 to various sesquiterpene lactones, clerodane and labdane diterpenoids, strychnine, and denatonium.

    Science.gov (United States)

    Brockhoff, Anne; Behrens, Maik; Massarotti, Alberto; Appendino, Giovanni; Meyerhof, Wolfgang

    2007-07-25

    Sesquiterpene lactones are a major class of natural bitter compounds occurring in vegetables and culinary herbs as well as in aromatic and medicinal plants, where they often represent the main gustatory and pharmacologically active component. Investigations on sesquiterpene lactones have mainly focused on their bioactive potential rather than on their sensory properties. In the present study, we report about the stimulation of heterologously expressed human bitter taste receptors, hTAS2Rs, by the bitter sesquiterpene lactone herbolide D. A specific response to herbolide D was observed i.a. for hTAS2R46, a so far orphan bitter taste receptor without any known ligand. By further investigation of its agonist pattern, we characterized hTAS2R46 as a bitter receptor broadly tuned to sesquiterpene lactones and to clerodane and labdane diterpenoids as well as to the unrelated bitter substances strychnine and denatonium.

  20. High-level diterpene production by transient expression in Nicotiana benthamiana

    Science.gov (United States)

    2013-01-01

    Background Characterization of plant terpene synthases is typically done by production of recombinant enzymes in Escherichia coli. This is often difficult due to solubility and codon usage issues. Furthermore, plant terpene synthases which are targeted to the plastids, such as diterpene synthases, have to be shortened in a more or less empirical approach to improve expression. We report here an optimized Agrobacterium-mediated transient expression assay in Nicotiana benthamiana for plant diterpene synthase expression and product analysis. Results Agrobacterium-mediated transient expression of plant diterpene synthases in N. benthamiana led to the accumulation of diterpenes within 3 days of infiltration and with a maximum at 5 days. Over 50% of the products were exported onto the leaf surface, thus considerably facilitating the analysis by reducing the complexity of the extracts. The robustness of the method was tested by expressing three different plant enzymes, cembratrien-ol synthase from Nicotiana sylvestris, casbene synthase from Ricinus communis and levopimaradiene synthase from Gingko biloba. Furthermore, co-expression of a 1-deoxy-D-xylulose-5-phosphate synthase from tomato and a geranylgeranyl diphosphate synthase from tobacco led to a 3.5-fold increase in the amount of cembratrien-ol produced, with maximum yields reaching 2500 ng/cm2. Conclusion With this optimized method for diterpene synthase expression and product analysis, a single infiltrated leaf of N. benthamiana would be sufficient to produce quantities required for the structure elucidation of unknown diterpenes. The method will also be of general use for gene function discovery, pathway reconstitution and metabolic engineering of diterpenoid biosynthesis in plants. PMID:24330621

  1. [Effects of ginkgo diterpene lactones meglumine injection's activated carbon adsorption technology on officinal components].

    Science.gov (United States)

    Zhou, En-li; Wang, Ren-jie; Li, Miao; Wang, Wei; Xu, Dian-hong; Hu, Yang; Wang, Zhen-zhong; Bi, Yu-an; Xiao, Wei

    2015-10-01

    With the diversion rate of ginkgolide A, B, K as comprehensive evaluation indexes, the amount of activated carbon, ad- sorption time, mix rate, and adsorption temperature were selected as factors, orthogonal design which based on the evaluation method of information entropy was used to optimize activated carbon adsorption technology of ginkgo diterpene lactones meglumine injection. Opti- mized adsorption conditions were as follows: adsorbed 30 min with 0.2% activated carbon in 25 °C, 40 r ·min⁻¹, validation test re- sult display. The optimum extraction condition was stable and feasible, it will provide a basis for ginkgo diterpene lactone meglumine injection' activated carbon adsorption process.

  2. A New Diterpene Glycoside: 15α-Hydroxy-Rebaudioside M Isolated from Stevia rebaudiana.

    Science.gov (United States)

    Prakash, Indra; Ma, Gil; Bunders, Cynthia; Devkota, Krishna P; Charan, Romila D; Ramirez, Catherine; Snyder, Tara M; Priedemann, Christopher

    2015-07-01

    In a continued search for novel diterpenoid glycosides, we recently isolated and characterized a Rebaudioside M derivative with a hydroxyl group at position 15 in the central diterpene core from an extract of Stevia rebaudiana Bertoni. Here we report the complete structure elucidation of 15α-hydroxy-Rebaudioside M (2) on the basis of NMR (1H, 13C, COSY, HSQC-DEPT, HMBC, 1D TOCSY, NOESY) and mass spectral data. Steviol glycoside with a hydroxyl group at C-15 in the central diterpene core has not been previously reported.

  3. HPLC-SPE-NMR for combinatorial biosynthetic investigations – expanding the landscape of diterpene structural diversity

    DEFF Research Database (Denmark)

    Kongstad, Kenneth Thermann; Andersen-Ranberg, Johan; Hamberger, Björn Robert

    In this work, the analytical technique, HPLC-HRMS-SPE-NMR was used for the first time in combination with combinatorial biosynthetic investigations in N. benthamiana. This efficient setup allowed for identification of several diterpene synthase (diTPS) combinations responsible for stereospecific ......In this work, the analytical technique, HPLC-HRMS-SPE-NMR was used for the first time in combination with combinatorial biosynthetic investigations in N. benthamiana. This efficient setup allowed for identification of several diterpene synthase (diTPS) combinations responsible for...

  4. Effects of kaurane diterpene derivatives on germination and growth of Lactuca sativa seedlings.

    Science.gov (United States)

    Vieira, Henriete S; Takahashi, Jacqueline A; Pimenta, Lúcia P S; Boaventura, Maria Amélia D

    2005-01-01

    Kaurenoic and grandiflorenic acid, isolated from Wedelia paludosa (Asteraceae), some derivatives from these acids (alcohols, esters, amides, lactones, oximes) and other naturally occurring kaurane diterpenes were tested for their action on the growth of radical and shoot of Lactuca sativa. Gibberellic acid, GA3, a commercially available phytohormone, belonging to the same class of diterpenes, was also tested. Some of the tested substances showed a remarkable activity either in the inhibition or in stimulation of L. sativa growth. The activity, in some cases, was even higher than that of GA3.

  5. Four gene products are required for the fungal synthesis of the indole-diterpene, paspaline.

    Science.gov (United States)

    Saikia, Sanjay; Parker, Emily J; Koulman, Albert; Scott, Barry

    2006-03-06

    Paspaline belongs to a large, structurally and functionally diverse group of indole-diterpenes synthesized by filamentous fungi. However, the identity of the gene products required for the biosynthesis of paspaline, a key intermediate for the synthesis of paxilline and other indole-diterpenes, is not known. Transfer of constructs containing different pax gene combinations into a paxilline negative deletion derivative of Penicillium paxilli demonstrated that just four proteins, PaxG, a geranylgeranyl diphosphate synthase, PaxM, a FAD-dependent monooxygenase, PaxB, a putative membrane protein, and PaxC, a prenyl transferase, are required for the biosynthesis of paspaline.

  6. Cordifolide A, a Sulfur-containing Clerodane Diterpene Glycoside from Tinospora cordifolia

    Science.gov (United States)

    Pan, Li; Terrazas, Cesar; Lezama-Davila, Claudio M.; Rege, Nirmala; Gallucci, Judith C.; Satoskar, Abhay R.; Kinghorn, A. Douglas

    2012-01-01

    Cordifolide A (1), a novel unprecedented sulfur-containing clerodane diterpene glycoside, together with other two new diterpene glycosides, cordifolides B (2) and C (3), and four known analogues, was isolated from a methanol-soluble extract of the stems of Tinospora cordifolia. The structures of the new compounds were determined on the basis of spectroscopic data interpretation, with that of cordifolide A (1) confirmed by a single-crystal X-ray crystallographic analysis. All isolates were evaluated for their in vitro immunomodulatory activity using mouse bone marrow-derived dentritic cells (BMDCs). PMID:22497272

  7. Enzymatic 13C Labeling and Multidimensional NMR Analysis of Miltiradiene Synthesized by Bifunctional Diterpene Cyclase in Selaginella moellendorffii*

    Science.gov (United States)

    Sugai, Yoshinori; Ueno, Yohei; Hayashi, Ken-ichiro; Oogami, Shingo; Toyomasu, Tomonobu; Matsumoto, Sadamu; Natsume, Masahiro; Nozaki, Hiroshi; Kawaide, Hiroshi

    2011-01-01

    Diterpenes show diverse chemical structures and various physiological roles. The diversity of diterpene is primarily established by diterpene cyclases that catalyze a cyclization reaction to form the carbon skeleton of cyclic diterpene. Diterpene cyclases are divided into two types, monofunctional and bifunctional cyclases. Bifunctional diterpene cyclases (BDTCs) are involved in hormone and defense compound biosyntheses in bryophytes and gymnosperms, respectively. The BDTCs catalyze the successive two-step type-B (protonation-initiated cyclization) and type-A (ionization-initiated cyclization) reactions of geranylgeranyl diphosphate (GGDP). We found that the genome of a lycophyte, Selaginella moellendorffii, contains six BDTC genes with the majority being uncharacterized. The cDNA from S. moellendorffii encoding a BDTC-like enzyme, miltiradiene synthase (SmMDS), was cloned. The recombinant SmMDS converted GGDP to a diterpene hydrocarbon product with a molecular mass of 272 Da. Mutation in the type-B active motif of SmMDS abolished the cyclase activity, whereas (+)-copalyl diphosphate, the reaction intermediate from the conversion of GGDP to the hydrocarbon product, rescued the cyclase activity of the mutant to form a diterpene hydrocarbon. Another mutant lacking type-A activity accumulated copalyl diphosphate as the reaction intermediate. When the diterpene hydrocarbon was enzymatically synthesized from [U-13C6]mevalonate, all carbons were labeled with 13C stable isotope (>99%). The fully 13C-labeled product was subjected to 13C-13C COSY NMR spectroscopic analyses. The direct carbon-carbon connectivities observed in the multidimensional NMR spectra demonstrated that the hydrocarbon product by SmMDS is miltiradiene, a putative biosynthetic precursor of tanshinone identified from the Chinese medicinal herb Salvia miltiorrhiza. Hence, SmMDS functions as a bifunctional miltiradiene synthase in S. moellendorffii. In this study, we demonstrate that one-dimensional and

  8. Chemotaxonomy of three genera of the annonaceae family using self-organizing maps and 13C NMR data of diterpenes

    Directory of Open Access Journals (Sweden)

    Luciana Scotti

    2012-01-01

    Full Text Available The Annonaceae family is distributed throughout Neotropical regions of the world. In Brazil, it covers nearly all natural formations particularly Annona, Xylopia and Polyalthia and is characterized chemically by the production of sources of terpenoids (mainly diterpenes, alkaloids, steroids, polyphenols and, flavonoids. Studies from 13C NMR data of diterpenes related with their botanical occurrence were used to generate self-organizing maps (SOM. Results corroborate those in the literature obtained from morphological and molecular data for three genera and the model can be used to project other diterpenes. Therefore, the model produced can predict which genera are likely to contain a compound.

  9. A study of the diterpene alkaloids of Aconitum napellus ssp. neomontanum during its onthogenetic cycle.

    Science.gov (United States)

    Colombo, M L; Bravin, M; Tome, F

    1988-12-01

    Qualitative and quantitative variations in the diterpene alkaloids content in the neomontanum subspecies of Aconitum napellus, growing near lake of Como, were studied throughout its complete onthogenetic cycle. Aconitine, N-deethylaconitine and two compounds with m.w. = 688 and m.w. = 629 have been detected in the various organs of plant almost at any time during the vegetative cycle.

  10. Dolabelladienols A–C, New Diterpenes Isolated from Brazilian Brown Alga Dictyota pfaffii

    Directory of Open Access Journals (Sweden)

    Alonso Pardo-Vargas

    2014-07-01

    Full Text Available The marine brown alga Dictyota pfaffii from Atol das Rocas, in Northeast Brazil is a rich source of dolabellane diterpene, which has the potential to be used in future antiviral drugs by inhibiting reverse transcriptase (RT of HIV-1. Reexamination of the minor diterpene constituents yielded three new dolabellane diterpenes, (1R*,2E,4R*,7S,10S*,11S*,12R*10,18-diacetoxy-7-hydroxy-2,8(17-dolabelladiene (1, (1R*,2E,4R*,7R*,10S*,11S*,12R*10,18-diacetoxy-7-hydroxy-2,8(17-dolabelladiene (2, (1R*,2E,4R*,8E,10S*,11S,12R*10,18-diacetoxy-7-hydroxy-2,8-dolabelladiene (3, termed dolabelladienols A–C (1–3 respectively, in addition to the known dolabellane diterpenes (4–6. The elucidation of the compounds 1–3 was assigned by 1D and 2D NMR, MS, optical rotation and molecular modeling, along with the relative configuration of compound 4 and the absolute configuration of 5 by X-ray diffraction. The potent anti-HIV-1 activities displayed by compounds 1 and 2 (IC50 = 2.9 and 4.1 μM, which were more active than even the known dolabelladienetriol 4, and the low cytotoxic activity against MT-2 lymphocyte tumor cells indicated that these compounds are promising anti-HIV-1 agents.

  11. Dolabelladienols A–C, New Diterpenes Isolated from Brazilian Brown Alga Dictyota pfaffii

    Science.gov (United States)

    Pardo-Vargas, Alonso; Oliveira, Ingrid de Barcelos; Stephens, Paulo Roberto Soares; Cirne-Santos, Claudio Cesar; Paixão, Izabel Christina Nunes de Palmer; Ramos, Freddy Alejandro; Jiménez, Carlos; Rodríguez, Jaime; Resende, Jackson Antonio Lamounier Camargos; Teixeira, Valeria Laneuville; Castellanos, Leonardo

    2014-01-01

    The marine brown alga Dictyota pfaffii from Atol das Rocas, in Northeast Brazil is a rich source of dolabellane diterpene, which has the potential to be used in future antiviral drugs by inhibiting reverse transcriptase (RT) of HIV-1. Reexamination of the minor diterpene constituents yielded three new dolabellane diterpenes, (1R*,2E,4R*,7S,10S*,11S*,12R*)10,18-diacetoxy-7-hydroxy-2,8(17)-dolabelladiene (1), (1R*,2E,4R*,7R*,10S*,11S*,12R*)10,18-diacetoxy-7-hydroxy-2,8(17)-dolabelladiene (2), (1R*,2E,4R*,8E,10S*,11S,12R*)10,18-diacetoxy-7-hydroxy-2,8-dolabelladiene (3), termed dolabelladienols A–C (1–3) respectively, in addition to the known dolabellane diterpenes (4–6). The elucidation of the compounds 1–3 was assigned by 1D and 2D NMR, MS, optical rotation and molecular modeling, along with the relative configuration of compound 4 and the absolute configuration of 5 by X-ray diffraction. The potent anti-HIV-1 activities displayed by compounds 1 and 2 (IC50 = 2.9 and 4.1 μM), which were more active than even the known dolabelladienetriol 4, and the low cytotoxic activity against MT-2 lymphocyte tumor cells indicated that these compounds are promising anti-HIV-1 agents. PMID:25056631

  12. Dolabelladienols A-C, new diterpenes isolated from Brazilian brown alga Dictyota pfaffii.

    Science.gov (United States)

    Pardo-Vargas, Alonso; de Barcelos Oliveira, Ingrid; Stephens, Paulo Roberto Soares; Cirne-Santos, Claudio Cesar; de Palmer Paixão, Izabel Christina Nunes; Ramos, Freddy Alejandro; Jiménez, Carlos; Rodríguez, Jaime; Resende, Jackson Antonio Lamounier Camargos; Teixeira, Valeria Laneuville; Castellanos, Leonardo

    2014-07-23

    The marine brown alga Dictyota pfaffii from Atol das Rocas, in Northeast Brazil is a rich source of dolabellane diterpene, which has the potential to be used in future antiviral drugs by inhibiting reverse transcriptase (RT) of HIV-1. Reexamination of the minor diterpene constituents yielded three new dolabellane diterpenes, (1R*,2E,4R*,7S,10S*,11S*,12R*)10,18-diacetoxy-7-hydroxy-2,8(17)-dolabelladiene (1), (1R*,2E,4R*,7R*,10S*,11S*,12R*)10,18-diacetoxy-7-hydroxy-2,8(17)-dolabelladiene (2), (1R*,2E,4R*,8E,10S*,11S,12R*)10,18-diacetoxy-7-hydroxy-2,8-dolabelladiene (3), termed dolabelladienols A-C (1-3) respectively, in addition to the known dolabellane diterpenes (4-6). The elucidation of the compounds 1-3 was assigned by 1D and 2D NMR, MS, optical rotation and molecular modeling, along with the relative configuration of compound 4 and the absolute configuration of 5 by X-ray diffraction. The potent anti-HIV-1 activities displayed by compounds 1 and 2 (IC50 = 2.9 and 4.1 μM), which were more active than even the known dolabelladienetriol 4, and the low cytotoxic activity against MT-2 lymphocyte tumor cells indicated that these compounds are promising anti-HIV-1 agents.

  13. The effect of the diterpene 5-epi-icetexone on the cell cycle of Trypanosoma cruzi.

    NARCIS (Netherlands)

    Lozano, E.; Barrera, P.; Tonn, C.; Nieto, M.; Sartor, T.; Sosa, M.A.

    2012-01-01

    Numerous natural compounds have been used against Trypanosoma cruzi, the causative agent of Chagas' disease. Here, we studied the effect of the diterpene 5-epi-icetexone on growth and morphology of parasites synchronized with hydroxyurea, at different periods of time after removal of the nucleotide.

  14. Separate effects of the coffee diterpenes cafestol and kahweol on serum lipids and liver transaminases.

    NARCIS (Netherlands)

    Urgert, R.; Essed, N.; Weg, van der G.; Kosmeijer-Schuil, T.G.; Katan, M.B.

    1997-01-01

    The coffee diterpene cafestol occurs in both robusta and arabica beans. It is present in unfiltered coffee brews and raises serum concentrations of cholesterol, triacylglycerols, and alanine aminotransferase in humans. The effects are linear with the cafestol dose. Unfiltered coffee also contains

  15. The first structure of a bacterial diterpene cyclase: CotB2.

    Science.gov (United States)

    Janke, Ronja; Görner, Christian; Hirte, Max; Brück, Thomas; Loll, Bernhard

    2014-06-01

    Sesquiterpenes and diterpenes are a diverse class of secondary metabolites that are predominantly derived from plants and some prokaryotes. The properties of these natural products encompass antitumor, antibiotic and even insecticidal activities. Therefore, they are interesting commercial targets for the chemical and pharmaceutical industries. Owing to their structural complexity, these compounds are more efficiently accessed by metabolic engineering of microbial systems than by chemical synthesis. This work presents the first crystal structure of a bacterial diterpene cyclase, CotB2 from the soil bacterium Streptomyces melanosporofaciens, at 1.64 Å resolution. CotB2 is a diterpene cyclase that catalyzes the cyclization of the linear geranylgeranyl diphosphate to the tricyclic cyclooctat-9-en-7-ol. The subsequent oxidation of cyclooctat-9-en-7-ol by two cytochrome P450 monooxygenases leads to bioactive cyclooctatin. Plasticity residues that decorate the active site of CotB2 have been mutated, resulting in alternative monocyclic, dicyclic and tricyclic compounds that show bioactivity. These new compounds shed new light on diterpene cyclase reaction mechanisms. Furthermore, the product of mutant CotB2(W288G) produced the new antibiotic compound (1R,3E,7E,11S,12S)-3,7,18-dolabellatriene, which acts specifically against multidrug-resistant Staphylococcus aureus. This opens a sustainable route for the industrial-scale production of this bioactive compound.

  16. The effect of the diterpene 5-epi-icetexone on the cell cycle of Trypanosoma cruzi.

    NARCIS (Netherlands)

    Lozano, E.; Barrera, P.; Tonn, C.; Nieto, M.; Sartor, T.; Sosa, M.A.

    2012-01-01

    Numerous natural compounds have been used against Trypanosoma cruzi, the causative agent of Chagas' disease. Here, we studied the effect of the diterpene 5-epi-icetexone on growth and morphology of parasites synchronized with hydroxyurea, at different periods of time after removal of the nucleotide.

  17. Diterpenes from rosemary (Rosmarinus officinalis): Defining their potential for anti-cancer activity.

    Science.gov (United States)

    Petiwala, Sakina M; Johnson, Jeremy J

    2015-10-28

    Recently, rosemary extracts standardized to diterpenes (e.g. carnosic acid and carnosol) have been approved by the European Union (EU) and given a GRAS (Generally Recognized as Safe) status in the United States by the Food and Drug Administration (FDA). Incorporation of rosemary into our food system and through dietary selection (e.g. Mediterranean Diet) has increased the likelihood of exposure to diterpenes in rosemary. In consideration of this, a more thorough understanding of rosemary diterpenes is needed to understand its potential for a positive impact on human health. Three agents in particular have received the most attention that includes carnosic acid, carnosol, and rosmanol with promising results of anti-cancer activity. These studies have provided evidence of diterpenes to modulate deregulated signaling pathways in different solid and blood cancers. Rosemary extracts and the phytochemicals therein appear to be well tolerated in different animal models as evidenced by the extensive studies performed for approval by the EU and the FDA as an antioxidant food preservative. This mini-review reports on the pre-clinical studies performed with carnosic acid, carnosol, and rosmanol describing their mechanism of action in different cancers.

  18. Reconstitution of Biosynthetic Machinery for the Synthesis of the Highly Elaborated Indole Diterpene Penitrem

    DEFF Research Database (Denmark)

    Liu, Chengwei; Tagami, Koichi; Minami, Atsushi;

    2015-01-01

    Penitrem A is one of the most elaborated members of the fungal indole diterpenes. Two separate penitrem gene clusters were identified using genomic and RNA sequencing data, and 13 out of 17 transformations in the penitrem biosynthesis were elucidated by heterologous reconstitution of the relevant...

  19. A New Cembrane Diterpene from the Bornean Soft Coral Nephthea sp.

    Directory of Open Access Journals (Sweden)

    Takahiro Ishii

    2010-05-01

    Full Text Available A new cembrane diterpene, 6-acetoxy-7,8-epoxynephthenol acetate (1 was isolated along with a known compound, epoxynephthenol acetate (2, from the organic extract of a Bornean soft coral Nephthea sp. Their structures were elucidated on the basis of spectroscopic analyses and comparison with those previous literature data.

  20. Amitorines A and B, Nitrogenous Diterpene Metabolites of Theonella swinhoei: Isolation, Structure Elucidation, and Asymmetric Synthesis.

    Science.gov (United States)

    Ota, Koichiro; Hamamoto, Yukiko; Eda, Wakiko; Tamura, Kenta; Sawada, Akiyoshi; Hoshino, Ayako; Mitome, Hidemichi; Kamaike, Kazuo; Miyaoka, Hiroaki

    2016-04-22

    Two new nitrogenous prenylbisabolanes never before found in Lithistid sponges have been isolated from Theonella swinhoei. These new diterpenes, named amitorine A (1) and amitorine B (2), containing a prenylbisabolane skeleton have been characterized by spectroscopic analyses, and the relative and absolute configurations of 1 and 2 were determined by asymmetric synthesis of both diastereomers via the common bicyclic lactone 6 intermediate.

  1. Spiro Fused Diterpene-Indole Alkaloids from a Creek-Bottom-Derived Aspergillus terreus

    Science.gov (United States)

    Cai, Shengxin; Du, Lin; Gerea, Alexandra L.; King, Jarrod B.; You, Jianlan

    2013-01-01

    Four metabolites, teraspiridoles A–D (2–5), formed from the merger of diterpene and modified indole scaffold were obtained from an Aspergillus terreus isolate. The structures and absolute configurations of these natural products were established using NMR, mass spectrometry, Marfey’s method, VCD, and ECD data. Teraspiridole B (3) exhibited weak inhibition of planaria regeneration/survival. PMID:23924243

  2. Further New Cembrane Diterpenes from the Hainan Soft Coral Sarcophyton latum

    Institute of Scientific and Technical Information of China (English)

    YAN,Xiao-Hong; FENG,Lin-Yin; GUO,Yue-Wei

    2008-01-01

    Two new cembranoid diterpenes, diepoxysarcophytonene (1) and sarconphytonol (2), were isolated from the Hainan soft coral Sarcophyton latum. Their structures including relative stereochemistry were established by 1D and 2D NMR spectroscopic techniques. Compound 1 showed weak inhibitory activity against COX-2 in vitro at the concentration of 10-5 μmol·L-1 with the inhibitory rate of 55%.

  3. A New Cassane-type Diterpene from the Seed of Caesalpinia sappan.

    Science.gov (United States)

    Nguyen, Hai Xuan; Nguyen, Nhan Trung; Dang, Phu Hoang; Thi, Phuoc Ho; Nguyen, Mai Thanh Thi; Can, Mao Van; Dibwe, Dya Fita; Ueda, Jun-Ya; Matsumoto, Kinzo; Awale, Suresh

    2016-06-01

    Phytochemical investigation of the CH2Cl2 extract of the Vietnamese medicinal plant Caesalpinia sappan Linn resulted in the isolation of a new cassane-type diterpene named tomocin I (1). Its chemical structure was determined by NMR spectroscopic and mass spectrometric analysis.

  4. Synthesis of an Advanced Intermediate of the Jatrophane Diterpene Pl-4: A Dibromide Coupling Approach

    Science.gov (United States)

    2013-01-01

    The preparation of an advanced intermediate toward the synthesis of the jatrophane diterpene Pl-4 is described. The key step is a regioselective chelation-controlled lithiation of the (Z)-configured bromide in the corresponding vinyl dibromide precursor. The method outlined within this Article is suitable for the facile access of sterically hindered internal vinyl halides for further coupling reactions. PMID:23895274

  5. HPLC-SPE-NMR for combinatorial biosynthetic investigations – Expanding the landscape of diterpene structural diversity

    DEFF Research Database (Denmark)

    Kongstad, Kenneth Thermann; Andersen-Ranberg, Johan; Hamberger, Björn Robert;

    In this work, the analytical technique, HPLC-HRMS-SPE-NMR was used for the first time in combination with combinatorial biosynthetic investigations in N. benthamiana. This efficient setup allowed for identification of several diterpene synthase (diTPS) combinations responsible for stereospecific...

  6. Characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (Rosmarinus officinalis) glandular trichomes

    NARCIS (Netherlands)

    Brückner, K.; Bozic, D.; Manzano, D.; Papaefthimiou, D.; Pateraki, I.; Scheler, U.; Ferrer, A.; Vos, de R.C.H.; Kanellis, A.K.; Tissier, A.

    2014-01-01

    Rosemary (Rosmarinus officinalis) produces the phenolic diterpenes carnosic acid and carnosol, which, in addition to their general antioxidant activities, have recently been suggested as potential ingredients for the prevention and treatment of neurodegenerative diseases. Little is known about the b

  7. A single residue change leads to a hydroxylated product from the class II diterpene cyclization catalyzed by abietadiene synthase

    Science.gov (United States)

    Criswell, Jared; Potter, Kevin; Shephard, Freya; Beale, Michael H.; Peters, Reuben J.

    2012-01-01

    Class II diterpene cyclases catalyze bicyclization of geranylgeranyl diphosphate. While this reaction typically is terminated via methyl deprotonation to yield copalyl diphosphate, in rare cases hydroxylated bicycles are produced instead. Abietadiene synthase is a bifunctional diterpene cyclase that usually produces a copalyl diphosphate intermediate. Here it is shown that substitution of aspartate for a conserved histidine in the class II active site of abietadiene synthase leads to selective production of 8α-hydroxy-CPP instead, demonstrating striking plasticity. PMID:23167845

  8. New approaches on the analyses of thermolabile coffee diterpenes by gas chromatography and its relationship with cup quality.

    Science.gov (United States)

    Novaes, Fábio Junior Moreira; Oigman, Silvia Siag; de Souza, Rodrigo Octavio Mendonça Alves; Rezende, Claudia Moraes; de Aquino Neto, Francisco Radler

    2015-07-01

    A new gas chromatography method using pulsed split injector (PS-GC) was validated to quantify thermolabile diterpenes cafestol, kahweol and isokahweol in methanolysed Arabica coffee oils. Linearity was 0.99 from 8 to 69mgmL(-1), recovery ranged from 99% to 101% and precision of less than 4% was obtained. Besides, Soxhlet extraction time was evaluated and Tukey׳s test indicated that the mass of diterpenes obtained in 4h is equivalent to a 16h period, which represents a space-time yield four times higher. The microwave assisted methanolysis proved to be efficient to quantitatively convert the natural diterpene esters in their respective alcohols and fatty acid methyl esters, accompanied by PS-GC. Also, the intact diterpene esters were analyzed by GC for the first time by the comparison between cold on-column (COC) and PS injection techniques. In all these stages, the molecular integrity of the thermolabile furokaurane diterpenes was maintained. The methanolysed oils from 13 samples of green Brazilian Arabica coffees were analyzed by PS-GC and the diterpenes composition varied from 8 to 12% w/w in oil and 0.7-1% in coffee beans. The ratio between cafestol and kahweol was successfully used to predict the quality of coffee even before the roasting and brewing processes.

  9. Functional characterization of wheat ent-kaurene(-like) synthases indicates continuing evolution of labdane-related diterpenoid metabolism in the cereals.

    Science.gov (United States)

    Zhou, Ke; Xu, Meimei; Tiernan, Mollie; Xie, Qian; Toyomasu, Tomonobu; Sugawara, Chizu; Oku, Madoka; Usui, Masami; Mitsuhashi, Wataru; Chono, Makiko; Chandler, Peter M; Peters, Reuben J

    2012-12-01

    Wheat (Triticum aestivum) and rice (Oryza sativa) are two of the most agriculturally important cereal crop plants. Rice is known to produce numerous diterpenoid natural products that serve as phytoalexins and/or allelochemicals. Specifically, these are labdane-related diterpenoids, derived from a characteristic labdadienyl/copalyl diphosphate (CPP), whose biosynthetic relationship to gibberellin biosynthesis is evident from the relevant expanded and functionally diverse family of ent-kaurene synthase-like (KSL) genes found in rice the (OsKSLs). Herein reported is the biochemical characterization of a similarly expansive family of KSL from wheat (the TaKSLs). In particular, beyond ent-kaurene synthases (KS), wheat also contains several biochemically diversified KSLs. These react either with the ent-CPP intermediate common to gibberellin biosynthesis or with the normal stereoisomer of CPP that also is found in wheat (as demonstrated by the accompanying paper describing the wheat CPP synthases). Comparison with a barley (Hordeum vulgare) KS indicates conservation of monocot KS, with early and continued expansion and functional diversification of KSLs in at least the small grain cereals. In addition, some of the TaKSLs that utilize normal CPP also will react with syn-CPP, echoing previous findings with the OsKSL family, with such enzymatic promiscuity/elasticity providing insight into the continuing evolution of diterpenoid metabolism in the cereal crop plant family, as well as more generally, which is discussed here.

  10. Bacterial diterpene synthases: new opportunities for mechanistic enzymology and engineered biosynthesis.

    Science.gov (United States)

    Smanski, Michael J; Peterson, Ryan M; Huang, Sheng-Xiong; Shen, Ben

    2012-04-01

    Diterpenoid biosynthesis has been extensively studied in plants and fungi, yet cloning and engineering diterpenoid pathways in these organisms remain challenging. Bacteria are emerging as prolific producers of diterpenoid natural products, and bacterial diterpene synthases are poised to make significant contributions to our understanding of terpenoid biosynthesis. Here we will first survey diterpenoid natural products of bacterial origin and briefly review their biosynthesis with emphasis on diterpene synthases (DTSs) that channel geranylgeranyl diphosphate to various diterpenoid scaffolds. We will then highlight differences of DTSs of bacterial and higher organism origins and discuss the challenges in discovering novel bacterial DTSs. We will conclude by discussing new opportunities for DTS mechanistic enzymology and applications of bacterial DTS in biocatalysis and metabolic pathway engineering.

  11. Bioactive isopimarane diterpenes from the fungus, Epicoccum sp. HS-1, associated with Apostichopus japonicus.

    Science.gov (United States)

    Xia, Xuekui; Qi, Jun; Liu, Yayue; Jia, Airong; Zhang, Yonggang; Liu, Changheng; Gao, Cuiling; She, Zhigang

    2015-03-02

    One new isopimarane diterpene (1), together with two known compounds, 11-deoxydiaporthein A (2) and iso-pimara-8(14),15-diene (3) were isolated from the culture of Epicoccum sp., which was associated with Apostichopus japonicus. Their structures were determined by the analysis of 1D and 2D NMR, as well as mass spectroscopic data. The absolute configuration of Compound 1 was deduced by a single-crystal X-ray diffraction experiment using CuKα radiation. In the bioactivity assay, both Compounds 1 and 2 exhibited α-glucosidase inhibitory activity with IC50 values of 4.6 ± 0.1 and 11.9 ± 0.4 μM, respectively. This was the first report on isopimarane diterpenes with α-glucosidase inhibitory activity.

  12. Guaiane sesquiterpenes and isopimarane diterpenes from an endophytic fungus Xylaria sp.

    Science.gov (United States)

    Wu, Shao-Hua; He, Jian; Li, Xiao-Nian; Huang, Rong; Song, Fei; Chen, You-Wei; Miao, Cui-Ping

    2014-09-01

    Nine oxygenated guaiane-type sesquiterpenes and three isopimarane diterpenes were isolated from the culture broth of an endophytic fungus, Xylaria sp. YM 311647, obtained from Azadirachta indica. The structures of these compounds were elucidated by interpretation of spectroscopic data. The absolute configurations of two of these were confirmed by X-ray crystallographic analysis. All of the compounds were tested for their antifungal activities against five pathogenic fungal cells. The results showed that nine sesquiterpenes were moderately active against Candida albicans and Hormodendrum compactum with MIC values ranging from 32 to 256μg/ml, while the diterpenes were more active; One of these exhibited the most potent inhibitory activity against C. albicans and Pyricularia oryzae with MIC values of 16μg/ml.

  13. Mode of Action of Diterpene and Characterization of Related Metabolites from the Soft Coral, Xenia elongata

    Science.gov (United States)

    Andrianasolo, Eric H.; Haramaty, Liti; White, Eileen; Lutz, Richard; Falkowski, Paul

    2014-01-01

    Chemical and biological investigation of the cultured marine soft coral Xenia elongata led to the isolation of two new diterpenes (2, 3). Their structures were elucidated using a combination of NMR and mass spectrometry. Biological evaluations and assessments were determined using the specific apoptosis induction assay based on genetically engineered mammalian cell line D3 deficient in Bak and Bax and derived from a mouse epithelial cell. The diterpenes induce apoptosis in low micromolar concentrations. The results indicate that the previously isolated compound (1) affects cell in a manner similar to that of HSP90 and HDAC inhibitors and in a manner opposite of PI3 kinase/mTOR inhibitors. Compound (3) inhibits selectively HDAC6 in high micromolar concentrations. PMID:24562393

  14. Casbane Diterpene as a Promising Natural Antimicrobial Agent against Biofilm-Associated Infections

    Directory of Open Access Journals (Sweden)

    Edson Holanda Teixeira

    2010-12-01

    Full Text Available Croton nepetaefolius is a native plant from northeastern Brazil that belongs to the Euphorbiaceae family. The biological action of this plant has been extensively explored, being the secondary metabolites responsible for its properties alkaloids, diterpenes, and triterpenes. This study aimed to evaluate the ability of casbane diterpene (CD, isolated from the ethanolic extract of C. nepetaefolius, to inhibit microbial growth and biofilm formation of several clinical relevant species (bacteria and yeasts. It was found that CD possessed biocidal and biostatic activity against the majority of the species screened, with minimal active concentrations ranging between 125 and 500 µg/mL. In addition, it was observed that biofilm formation was inhibited even when the planktonic growth was not significantly affected. In conclusion, CD showed potential to be a natural tool for the treatment of diseases caused by different infectious microorganisms.

  15. Bacterial Diterpene Synthases: New Opportunities for Mechanistic Enzymology and Engineered Biosynthesis

    Science.gov (United States)

    Smanski, Michael J.; Peterson, Ryan M.; Huang, Sheng-Xiong; Shen, Ben

    2012-01-01

    Diterpenoid biosynthesis has been extensively studied in plants and fungi, yet cloning and engineering diterpenoid pathways in these organisms remain challenging. Bacteria are emerging as prolific producers of diterpenoid natural products, and bacterial diterpene synthases are poised to make significant contributions to our understanding of terpenoid biosynthesis. Here we will first survey diterpenoid natural products of bacterial origin and briefly review their biosynthesis with emphasis on diterpene synthases (DTSs) that channel geranylgeranyl diphosphate to various diterpenoid scaffolds. We will then highlight differences of DTSs of bacterial and higher organism origins and discuss the challenges in discovering novel bacterial DTSs. We will conclude by discussing new opportunities for DTS mechanistic enzymology and applications of bacterial DTS in biocatalysis and metabolic pathway engineering. PMID:22445175

  16. Mode of Action of Diterpene and Characterization of Related Metabolites from the Soft Coral, Xenia elongata

    Directory of Open Access Journals (Sweden)

    Eric H. Andrianasolo

    2014-02-01

    Full Text Available Chemical and biological investigation of the cultured marine soft coral Xenia elongata led to the isolation of two new diterpenes (2, 3. Their structures were elucidated using a combination of NMR and mass spectrometry. Biological evaluations and assessments were determined using the specific apoptosis induction assay based on genetically engineered mammalian cell line D3 deficient in Bak and Bax and derived from a mouse epithelial cell. The diterpenes induce apoptosis in low micromolar concentrations. The results indicate that the previously isolated compound (1 affects cell in a manner similar to that of HSP90 and HDAC inhibitors and in a manner opposite of PI3 kinase/mTOR inhibitors. Compound (3 inhibits selectively HDAC6 in high micromolar concentrations.

  17. A New Atisane-Type Diterpene from the Bark of the Mangrove Plant Excoecaria Agallocha

    Directory of Open Access Journals (Sweden)

    Zhan Chang Wang

    2009-01-01

    Full Text Available A new atisane-type diterpene, ent-16α-hydroxy-atisane-3,4-lactone (4 and three known diterpenes, ent-16α-hydroxy-atisane-3-one (1, ent-atisane-3β,16α-diol (2, ent-3,4-seco-16α-hydroxyatis- 4(19-en -3-oic acid (3 were isolated from the bark of the mangrove plant Excoecaria agallocha. Their structures and relative stereochemistry were elucidated by means of extensive NMR and MS analysis. Compound 3 exhibited significant anti-microfouling activity against the adherence of Pseudomonas pseudoalcaligenes, with an EC50 value of 0.54 ± 0.01 ppm.

  18. Diterpenes Synthesized from the Natural Serrulatane Leubethanol and Their in Vitro Activities against Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Ricardo Escarcena

    2015-04-01

    Full Text Available Seventeen new derivatives of the natural diterpene leubethanol, including some potential pro-drugs, with changes in the functionality of the aliphatic chain or modifications of aromatic ring and the phenolic group, were synthesized and tested in vitro by the MABA technique for their activity against the H37Rv strain of Mycobacterium tuberculosis. Some compounds showed antimycobacterial selectivity indices higher than leubethanol.

  19. Mode of antibacterial action of totarol, a diterpene from Podocarpus nagi.

    Science.gov (United States)

    Haraguchi, H; Oike, S; Muroi, H; Kubo, I

    1996-04-01

    The antimicrobial mechanism of totarol was studied using Pseudomonas aeruginosa IFO 3080. This diterpene inhibited oxygen consumption and respiratory-driven proton translocation in whole cells, and oxidation of NADH in membrane preparation. NADH-cytochrome c reductase was inhibited by totarol while cytochrome c oxidase was not. NADH-DPIP reductase and NADH-CoQ reductase were also inhibited. The site of respiratory inhibition of totarol was thought to be near CoQ in the bacterial electron transport chain.

  20. Palladium-catalyzed transformations of salvinorin A, a neoclerodane diterpene from Salvia divinorum.

    Science.gov (United States)

    Riley, Andrew P; Day, Victor W; Navarro, Hernán A; Prisinzano, Thomas E

    2013-12-06

    Transformations that selectively modify the furan ring present in a variety of naturals products would be useful in the synthesis of biological probes but remain largely underexplored. The neoclerodane diterpene salvinorin A, isolated from Salvia divinorum, is an example of a furan-containing natural product. Following selective bromination of salvinorin A, Suzuki-Miyaura and Sonogashira couplings were accomplished in moderate to good yields without hydrolyzing the labile C-2 acetate or altering the stereochemistry of the epimerizable centers.

  1. Clerodane type diterpene as a novel antifungal agent from Polyalthia longifolia var. pendula.

    Science.gov (United States)

    Bhattacharya, Asish K; Chand, Hemender R; John, Jyothis; Deshpande, Mukund V

    2015-04-13

    Bioactivity-guided chemical examination of methanolic extract of leaves of Polyalthia longifolia var. pendula led to the isolation of the active constituent, a diterpene 1 which was identified as 16α-hydroxycleroda-3,13(14)Z-dien-15,16-olide on the basis of its spectral data. Among the tested strains, diterpene 1 was found to exhibit antifungal activities having MIC90 values of 50.3, 100.6 and 201.2 μM against Candida albicans NCIM3557, Cryptococcus neoformans NCIM3542 (human pathogens) and Neurospora crassa NCIM870 (saprophyte), respectively. Initial, structure-activity-relationship (SAR) data generated by synthesizing some derivatives revealed that the double bond between C3-C4 and the free hydroxyl group at C16 are crucial for the antifungal activity of the diterpene 1. The mode of action of 1 in C. albicans is due to compromised cell membrane permeability and also probably due to disruption of cell wall structures. The red blood cell haemolysis of all the compounds (1-4) did not show any significant haemolysis and was found to be less than 15% for all the compounds when tested at highest concentration, i.e. 1200 μM. Interestingly, all the tested compounds inhibited Y-H transition in dimorphic C. albicans NCIM3557 at much lower concentration than their MIC90 values. Determination of ROS generation by diterpene 1 using DCFH-DA and DHR123 (dihydrorhodamine) staining of C. albicans NCIM3557 indicated production of intracellular ROS as a mechanism of antifungal activity.

  2. Bioactivity of Diterpens from the Ethyl Acetate Extract of Kingiodendron pinnatum Rox. Hams

    Science.gov (United States)

    Javarappa, Komal Kumar; Prasad, Attemode Girijanna Devi; Mahadesh Prasad, AJ; Mane, Chetana

    2016-01-01

    Background: Kingiodendron pinnatum Rox. Hams. is an endangered medicinal plant used in gonorrhoe, catarrhal conditions of genito-urinary and respiratory tracts. The scientific and pharmacological formulation of K. pinnatum has not been established so far though it is being traditionally used by tribes of the region. Objective: P hytochemical screening and identification of the bioactive compounds from the ethyl acetate extract of Kingiodendron pinnatum Rox. Hams. Materials and Methods: Chromatographic separation was carried out by thin layer chromatography and column chromatography. Bio-autography of the column fractioned extract and TLC chromatogram were evaluated in vitro for antibacterial activity. The PTLC, HP TLC were used for crude extract and HPLC, LCMS, FTIR, 1HNMR and 13CNMR were employed for the isolated compound in the ethyl acetate extract of K. pinnatum. Results: Evaluation of solvent system for chromatographic separation revealed that ethyl acetate: petroleum ether in the ratio of 7:2.5 ml was the most appropriate one for the separation of diterpene compounds. The antibacterial bio-autography screening of TLC separated compound showed positive activity with Staphylococcus aureus and negative activity with Escherichia coli. Spectroscopic analysis of the isolated compound from the ethyl acetate extract of K. pinnatum revealed the presence of diterpene compound. Conclusion: It is evident from the present study that the ethyl acetate extract of K. pinnatum is rich in diterpene compounds and having potential antibacterial activity. SUMMARY Novel extraction method for phytochemicls from Kingidendron pinnatum at RTAntibacterial property of diterpens extracted from Kingiodendron pinnatum Rox. Hams aganist S. aureus Abbreviations Used: TLC: Thin Layer Chromatography, PTLC: Preparatory Thin Layer Chromatography, HPTLC: High perormence Thin Layer chromatography, HPLC: High Performance Liquid Chromatography, LC-MS: Liquid chromatography Mass Spectra, FTIR

  3. Four New Cembrane Diterpenes from the Hainan Soft Coral Lobophytum sp.

    Institute of Scientific and Technical Information of China (English)

    CHEN,Si-Han; HUANG,Hui; GUO,Yue-Wei

    2008-01-01

    Four new cembrane diterpenes, 19-hydroxy-sarcocrassolide (1), 18-deacetyldeepoxy lobolide (2), 13-hydroxy-sinularial A (3) and 16-hydroxy-sinulariol C (4), along with four related known compounds (5-8), were isolated from the Hainan soft coral Lobophytum sp. Their structures, including relative stereochemistry, were elucidated by detailed analyses of spectroscopic data and by comparison with the data reported in literature.

  4. 13-Epi-9-deacetoxyxenicin, a Cytotoxic Diterpene from the Soft Coral Asterospicularia laurae (Alcyonacea

    Directory of Open Access Journals (Sweden)

    Bruce F. Bowden

    2003-11-01

    Full Text Available Abstract: An investigation of the soft coral Asterospicularia laurae collected on the Great Barrier Reef, Australia, afforded the cytotoxic diterpene 13-epi-9-deacetoxyxenicin (1 in addition to the known metabolites 13-epi-9-deacetylxenicin (2 and gorgosterol. 13-Epi-9-deacetoxyxenicin readily underwent an autoxidation reaction in solution to afford a single product, the hydroperoxide 3. Structures, stereochemistry, and NMR assignments were established by high resolution NMR spectroscopy and by comparison with published data for known compounds.

  5. Palladium-catalyzed Transformations of Salvinorin A, a Neoclerodane Diterpene from Salvia divinorum

    Science.gov (United States)

    Riley, Andrew P.; Day, Victor W.; Navarro, Hernán A.

    2013-01-01

    Transformations that selectively modify the furan ring present in a variety of naturals products would be useful in the synthesis of biological probes but remain largely underexplored. The neoclerodane diterpene salvinorin A, isolated from Salvia divinorum, is an example of a furan-containing natural product. Following selective bromination of salvinorin A, Suzuki-Miyaura and Sonogashira couplings were accomplished in moderate to good yields without hydrolyzing the labile C-2 acetate or altering the stereochemistry of the epimerizable centers. PMID:24246026

  6. Hepatoprotective prenylaromadendrane-type diterpenes from the gum resin of Boswellia carterii.

    Science.gov (United States)

    Wang, Yan-gai; Ren, Jin; Wang, Ai-guo; Yang, Jian-bo; Ji, Teng-fei; Ma, Qin-Ge; Tian, Jin; Su, Ya-lun

    2013-11-22

    Chemical examination of the exuded gum resin of Boswellia carterii resulted in the isolation of nine new prenylaromadendrane-type diterpenes, boscartols A-I (1-9). The structures of these compounds were established by extensive 1D and 2D NMR spectroscopic analyses, mass spectrometric data, and circular dichroism spectra. Compounds 1-3, 5, 6, 8, and 9 (10 μM) showed moderate hepatoprotective activity against d-galactosamine-induced HL-7702 cell damage.

  7. Efficient diterpene production in yeast by engineering Erg20p into a geranylgeranyl diphosphate synthase.

    Science.gov (United States)

    Ignea, Codruta; Trikka, Fotini A; Nikolaidis, Alexandros K; Georgantea, Panagiota; Ioannou, Efstathia; Loupassaki, Sofia; Kefalas, Panagiotis; Kanellis, Angelos K; Roussis, Vassilios; Makris, Antonios M; Kampranis, Sotirios C

    2015-01-01

    Terpenes have numerous applications, ranging from pharmaceuticals to fragrances and biofuels. With increasing interest in producing terpenes sustainably and economically, there has been significant progress in recent years in developing methods for their production in microorganisms. In Saccharomyces cerevisiae, production of the 20-carbon diterpenes has so far proven to be significantly less efficient than production of their 15-carbon sesquiterpene counterparts. In this report, we identify the modular structure of geranylgeranyl diphosphate synthesis in yeast to be a major limitation in diterpene yields, and we engineer the yeast farnesyl diphosphate synthase Erg20p to produce geranylgeranyl diphosphate. Using a combination of protein and genetic engineering, we achieve significant improvements in the production of sclareol and several other isoprenoids, including cis-abienol, abietadiene and β-carotene. We also report the development of yeast strains carrying the engineered Erg20p, which support efficient isoprenoid production and can be used as a dedicated chassis for diterpene production or biosynthetic pathway elucidation. The design developed here can be applied to the production of any GGPP-derived isoprenoid and is compatible with other yeast terpene production platforms.

  8. Kaurane and pimarane-type diterpenes from the Viguiera species inhibit vascular smooth muscle contractility.

    Science.gov (United States)

    Ambrosio, Sergio R; Tirapelli, Carlos R; da Costa, Fernando B; de Oliveira, Ana M

    2006-08-01

    The research, development and use of natural products as therapeutic agents, especially those derived from plants, have been increasing in recent years. Despite the fact that plants provide a rich source of novel biologically active compounds, only a small percentage have been phytochemically investigated and studied for their medical potential. Viguiera is a genus that belongs to the family Asteraceae and to the sunflower tribe Heliantheae, which is widespread mostly in Mexico and in other areas of the Andes and upland areas of Brazil. A review on the secondary metabolites pointed out that sesquiterpene lactones and diterpenes, of the kaurane and pimarane-type, are the main compounds produced by these plants. Some reports have shown that kaurane- and pimarane-type diterpenes exert several biological activities such as anti-inflammatory action, antimicrobial and antispasmodic activities. Kaurenoic and pimaradienoic acids, which are the main secondary metabolites isolated by our research group from the roots of Viguiera robusta and V. arenaria, respectively, have been evaluated on vascular smooth muscle contractility. We showed that these diterpenoids are able to inhibit the vascular contractility mainly by blocking extracellular Ca(2+) influx. Additionally, in this review we discuss the structure-activity relationship of the diterpenes regarding their inhibitory activity on vascular contractility.

  9. Cytotoxic diterpenes from roots of Crossopetalum gaumeri, a Celastraceae species from Yucatan Peninsula.

    Science.gov (United States)

    Miron-Lopez, Gumersindo; Bazzocchi, Isabel L; Jimenez-Diaz, Ignacio A; Moujir, Laila M; Quijano-Quiñones, Ramiro; Quijano, Leovigildo; Mena-Rejon, Gonzalo J

    2014-05-01

    Four new diterpenes, crossogumerins A-D (1-4) along with six known ones (5-10) were isolated from the root bark of Crossopetalum gaumeri, an endemic medicinal plant from the Yucatan Peninsula. Their structures were elucidated on the basis of 1D and 2D NMR techniques, including HMQC, HMBC, and ROESY experiments. Compounds 1-5, 8-10 were evaluated for cytotoxicity against HeLa (carcinoma of the cervix) and Hep-2 (lung carcinoma) human tumor cells lines and against normal Vero cells (African green monkey kidney) in lag and log phase of growth. Podocarpane diterpenes, crossogumerin B (2) and nimbiol (10), exhibited the highest activity against HeLa cells (IC50 values of 3.1 and 8.1 μM, respectively), but also selectivity on Vero cells (SI 22.6 and 7.5, respectively). The preliminary SAR studies suggest that an epoxy moiety in ring B and a hydrogen bond-donor group strategically positioned in the diterpene core are important requirements for cytotoxicity and selectivity.

  10. A Novel Diterpene Glycoside with Nine Glucose Units from Stevia rebaudiana Bertoni

    Directory of Open Access Journals (Sweden)

    Indra Prakash

    2017-01-01

    Full Text Available Following our interest in new diterpene glycosides with better taste profiles than that of Rebaudioside M, we have recently isolated and characterized Rebaudioside IX—a novel steviol glycoside—from a commercially‐supplied extract of Stevia rebaudiana Bertoni. This molecule contains a hexasaccharide group attached at C‐13 of the central diterpene core, and contains three additional glucose units when compared with Rebaudioside M. Here we report the complete structure elucidation—based on extensive Nuclear Magnetic Resonance (NMR analysis (1H, 13C, Correlation Spectroscopy (COSY, Heteronuclear Single Quantum Coherence‐Distortionless Enhancement Polarization Transfer (HSQC‐DEPT, Heteronuclear Multiple Bond Correlation (HMBC, 1D Total Correlation Spectroscopy (TOCSY, Nuclear Overhauser Effect Spectroscopy (NOESY and mass spectral data—of this novel diterpene glycoside with nine sugar moieties and containing a relatively rare 16 α‐linked glycoside. A steviol glycoside bearing nine glucose units is unprecedented in the literature, and could have an impact on the natural sweetener catalog.

  11. A Novel Diterpene Glycoside with Nine Glucose Units from Stevia rebaudiana Bertoni

    Science.gov (United States)

    Prakash, Indra; Ma, Gil; Bunders, Cynthia; Charan, Romila D.; Ramirez, Catherine; Devkota, Krishna P.; Snyder, Tara M.

    2017-01-01

    Following our interest in new diterpene glycosides with better taste profiles than that of Rebaudioside M, we have recently isolated and characterized Rebaudioside IX—a novel steviol glycoside—from a commercially-supplied extract of Stevia rebaudiana Bertoni. This molecule contains a hexasaccharide group attached at C-13 of the central diterpene core, and contains three additional glucose units when compared with Rebaudioside M. Here we report the complete structure elucidation—based on extensive Nuclear Magnetic Resonance (NMR) analysis (1H, 13C, Correlation Spectroscopy (COSY), Heteronuclear Single Quantum Coherence-Distortionless Enhancement Polarization Transfer (HSQC-DEPT), Heteronuclear Multiple Bond Correlation (HMBC), 1D Total Correlation Spectroscopy (TOCSY), Nuclear Overhauser Effect Spectroscopy (NOESY)) and mass spectral data—of this novel diterpene glycoside with nine sugar moieties and containing a relatively rare 1→6 α-linked glycoside. A steviol glycoside bearing nine glucose units is unprecedented in the literature, and could have an impact on the natural sweetener catalog. PMID:28146121

  12. ent-Kaurane diterpenes from the stem bark of Annona vepretorum (Annonaceae) and cytotoxic evaluation.

    Science.gov (United States)

    Dutra, Lívia M; Bomfim, Larissa M; Rocha, Suellen L A; Nepel, Angelita; Soares, Milena B P; Barison, Andersson; Costa, Emmanoel V; Bezerra, Daniel P

    2014-08-01

    This work describes a novel ent-kaurane diterpene, ent-3β-hydroxy-kaur-16-en-19-al along with five known ent-kaurane diterpenes, ent-3β,19-dihydroxy-kaur-16-eno, ent-3β-hydroxy-kaur-16-eno, ent-3β-acetoxy-kaur-16-eno, ent-3β-hydroxy-kaurenoic acid and kaurenoic acid, as well as caryophyllene oxide, humulene epoxide II, β-sitosterol, stigmasterol and campesterol from the stem bark of Annona vepretorum Mart. (Annonaceae). Cytotoxic activities towards tumor B16-F10, HepG2, K562 and HL60 and non-tumor PBMC cell lines were evaluated for ent-kaurane diterpenes. Among them, ent-3β-hydroxy-kaur-16-en-19-al was the most active compound with higher cytotoxic effect over K562 cell line (IC50 of 2.49 μg/mL) and lower over B16-F10 cell line (IC50 of 21.02 μg/mL).

  13. Two new diterpene derivatives from Euphorbia lunulata Bge and their anti-proliferative activities.

    Science.gov (United States)

    Liu, Chao; Liao, Zhi-xin; Liu, Shi-jun; Qu, Yan-bo; Wang, Heng-shan

    2014-07-01

    A new ent-abietane-type diterpene lactone (1) and a new jatrophane-type diterpenoid (2), together with twelve known compounds including three diterpenes (3-5), five triterpenes (6-10) and four sterides (11-14) were isolated from the ethanol extract of the whole plant of Euphorbia lunulata Bge. The structure of compounds 1 and 2 was elucidated on the basis of 1D and 2D NMR spectra and the HR-ESI-MS data. The structure of compound 2 was further analyzed by an X-ray crystallographic study. The in vitro anti-proliferative activities against MCF-7 and NCI-H460 cell lines for compounds 1-5 (diterpene) were evaluated. The results showed marked activity for compound 1 against the two cell lines with the IC50 values 19.5 (NCI-H460) and 18.6 (MCF-7) μM, while for cis-platinum (a positive cytotoxic control agent) 29.7 (NCI-H460) and 27.7 (MCF-7) μM. Compounds 2-5 exhibited moderate cytotoxic activities for the two cell lines with the IC50 values ranging from 32.1 to 58.2 μM.

  14. Antiviral Activity of Diterpene Esters on Chikungunya Virus and HIV Replication.

    Science.gov (United States)

    Nothias-Scaglia, Louis-Félix; Pannecouque, Christophe; Renucci, Franck; Delang, Leen; Neyts, Johan; Roussi, Fanny; Costa, Jean; Leyssen, Pieter; Litaudon, Marc; Paolini, Julien

    2015-06-26

    Recently, new daphnane, tigliane, and jatrophane diterpenoids have been isolated from various Euphorbiaceae species, of which some have been shown to be potent inhibitors of chikungunya virus (CHIKV) replication. To further explore this type of compound, the antiviral activity of a series of 29 commercially available natural diterpenoids was evaluated. Phorbol-12,13-didecanoate (11) proved to be the most potent inhibitor, with an EC50 value of 6.0 ± 0.9 nM and a selectivity index (SI) of 686, which is in line with the previously reported anti-CHIKV potency for the structurally related 12-O-tetradecanoylphorbol-13-acetate (13). Most of the other compounds exhibited low to moderate activity, including an ingenane-type diterpene ester, compound 28, with an EC50 value of 1.2 ± 0.1 μM and SI = 6.4. Diterpene compounds are known also to inhibit HIV replication, so the antiviral activities of compounds 1-29 were evaluated also against HIV-1 and HIV-2. Tigliane- (4β-hydroxyphorbol analogues 10, 11, 13, 15, 16, and 18) and ingenane-type (27 and 28) diterpene esters were shown to inhibit HIV replication in vitro at the nanomolar level. A Pearson analysis performed with the anti-CHIKV and anti-HIV data sets demonstrated a linear relationship, which supported the hypothesis made that PKC may be an important target in CHIKV replication.

  15. An abietane diterpene from Salvia cuspidata and some new derivatives are active against Trypanosoma cruzi.

    Science.gov (United States)

    Lozano, E S; Spina, R M; Tonn, C E; Sosa, M A; Cifuente, D A

    2015-12-01

    The Plant Kingdom is an excellent source for obtaining natural compounds with antiprotozoal activity. In the present work, we studied the effect of the diterpene 12-hydroxy-11,14-diketo-6,8,12-abietatrien-19,20-olide (HABTO) obtained from the aerial parts of Salvia cuspidata on Trypanosoma cruzi epimastigotes. This compound was found to inhibit parasite growth even at low concentrations (IC50 5 μg/mL) and with low toxicity on mammalian cells. In addition, this diterpene induced an intense vacuolization within the parasites. In order to obtain analogs with greater lipophilicity, chemical modifications on the enol moiety were carried out to obtain the acetyl (AABTO), the sylil (SABTO) and the allyl (ALLABTO) derivatives. We observed that the SABTO was the most effective one on the parasites, and the effect could be attributed to a greater lipophilicity of this compound. Taking into account these data we conclude that the increase of lipophilicity by chemical modifications is an adequate strategy for improving the trypanocidal activity of this kind abietane diterpenes.

  16. Pimarane Diterpenes and a Sesquiterpene from Salzmmania nitida

    Directory of Open Access Journals (Sweden)

    Mário G. de Carvalho

    2006-03-01

    Full Text Available Two new terpenoids, (+-3-oxo-thermarol and 11-acetoxyeudesman-4alpha-methyl-5alpha-ol along with the (+- thermarol were isolated from the aerial parts of Salzmmania nitida. The structures and unambiguous ¹H and 13C chemical shift assignments were established by spectroscopic means, including homo and heteronuclear techniques.O estudo fitoquímico de Salzmmania nitida D.C. (Rubiaceae conduziu ao isolamento e identificação de dois novos terpenoides, (+-3-oxo-thermarol e 11-acetoxi-4alfa-metil-5alfa-eudesmanol além do (+-termarol. As estruturas foram estabelecidas com base na análise de espectros de IV, Massas e RMN de ¹H e 13C (1D e 2D.

  17. 15-O-Acetyl-3-O-benzoylcharaciol and helioscopinolide A, two diterpenes isolated from Euphorbia helioscopia suppress microglia activation.

    Science.gov (United States)

    Wang, Hao; Liu, Yu; Zhang, Jingling; Xu, Jing; Cui, Chun-Ai; Guo, Yuanqiang; Jin, Da-Qing

    2016-01-26

    Microglia activation plays an important role in the pathogenesis of various neurodegenerative diseases by producing neurotoxic factors. In the present study, we found that two diterpenes isolated from Euphorbia helioscopia, 15-O-Acetyl-3-O-benzoylcharaciol and helioscopinolide A suppressed NO and PGE2 production by inhibition of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression in lipopolysaccharide (LPS)-stimulated BV2 microglia cells. The diterpenes also inhibited the production of ROS and proinflammatory cytokines including interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, the mechanism involved the NF-κB but not Akt and mitogen-activated protein kinase (MAPK) pathway. Moreover, the two diterpenes also attenuate microglia activation-mediated neuronal death. These results suggest that 15-O-Acetyl-3-O-benzoylcharaciol and helioscopinolide A may provide potential therapeutic strategy for various neuroinflammatory diseases.

  18. um estudo de caso

    OpenAIRE

    Costa, Cátia Filipa Pereira da

    2011-01-01

    Dissertação apresentada à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Psicologia Jurídica As situações de abuso sexual de crianças nas quais o perpetrador pertence ao sexo feminino obtiveram ao longo dos últimos anos um acrescido reconhecimento por parte da comunidade científica, evidenciado pelo significativo incremento das investigações no âmbito desta temática consistindo na sua maioria estudos de caso. Um conjunto de particularidades encont...

  19. Characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (Rosmarinus officinalis) glandular trichomes.

    Science.gov (United States)

    Brückner, Kathleen; Božić, Dragana; Manzano, David; Papaefthimiou, Dimitra; Pateraki, Irini; Scheler, Ulschan; Ferrer, Albert; de Vos, Ric C H; Kanellis, Angelos K; Tissier, Alain

    2014-05-01

    Rosemary (Rosmarinus officinalis) produces the phenolic diterpenes carnosic acid and carnosol, which, in addition to their general antioxidant activities, have recently been suggested as potential ingredients for the prevention and treatment of neurodegenerative diseases. Little is known about the biosynthesis of these diterpenes. Here we show that the biosynthesis of phenolic diterpenes in rosemary predominantly takes place in the glandular trichomes of young leaves, and used this feature to identify the first committed steps. Thus, a copalyl diphosphate synthase (RoCPS1) and two kaurene synthase-like (RoKSL1 and RoKSL2) encoding genes were identified and characterized. Expression in yeast (Saccharomyces cerevisiae) and Nicotiana benthamiana demonstrate that RoCPS1 converts geranylgeranyl diphosphate (GGDP) to copalyl diphosphate (CDP) of normal stereochemistry and that both RoKSL1 and RoKSL2 use normal CDP to produce an abietane diterpene. Comparison to the already characterized diterpene synthase from Salvia miltiorrhiza (SmKSL) demonstrates that the product of RoKSL1 and RoKSL2 is miltiradiene. Expression analysis supports a major contributing role for RoKSL2. Like SmKSL and the sclareol synthase from Salvia sclarea, RoKSL1/2 are diterpene synthases of the TPS-e group which have lost the internal gamma-domain. Furthermore, phylogenetic analysis indicates that RoKSL1 and RoKSL2 belong to a distinct group of KSL enzymes involved in specialized metabolism which most likely emerged before the dicot-monocot split.

  20. Anti-Angiogenic Properties of Cafestol and Kahweol Palmitate Diterpene Esters.

    Science.gov (United States)

    Moeenfard, Marzieh; Cortez, Alice; Machado, Vera; Costa, Raquel; Luís, Carla; Coelho, Pedro; Soares, Raquel; Alves, Arminda; Borges, Nuno; Santos, Alejandro

    2016-12-01

    Epidemiological studies support the association of coffee-specific diterpenes, with various beneficial health effects. Although anti-antiangiogenic properties of free cafestol and kahweol have been recently described, available data regarding their esterified form, in particular palmitate esters as the main diterpene esters present in coffee, are still rare. Given that angiogenesis plays an important role in many pathological conditions, including cancer growth and metastasis, this study aimed to assess and compare the potential anti-angiogenic effects of cafestol palmitate (CP) and kahweol palmitate (KP) in an in vitro angiogenesis model. According to our findings, both compounds inhibited angiogenesis steps on human microvascular endothelial cells (HMVECs), although a more significant effect was observed for KP. Compared to control, HMVECs viability decreased in a dose-dependent manner upon incubation either with CP or KP. Concentrations of 75 and 100 μM of each compound were cytotoxic. Cell proliferation was also dramatically reduced by both diterpene esters at 50 μM, although KP had a stronger inhibitory effect. However, CP and KP did not induce apoptosis on HMVECs. Both compounds reduced cell migration, but this effect was only statistically significant after KP incubation. Inhibition of VEGFR2 expression and its downstream effector Akt, but not Erk, was also observed in CP- and KP-treated HMVECs. These findings were confirmed using ELISA assay for phosphorylated (active) VEGFR-2. Taken together, these data indicate that both CP and KP can be considered potent compounds against angiogenesis-dependent disorders. Our findings further indicate that KP exerts more potent anti-angiogenic effects than CP, in most of assays. J. Cell. Biochem. 117: 2748-2756, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  1. Two novel aconane-type diterpenes derived from C-nor-C19-diterpenoid alkaloid

    Institute of Scientific and Technical Information of China (English)

    Xiang Li Shen; Qiao Hong Chen; Dong Lin Chen; Feng Peng Wang

    2009-01-01

    Two novel C-nor-B-homo aconane-type diterpenes 4 and 5.featuring a unique eight-membered ring system,were obtained by the treatment of C-nor-C19-diterpenoid alkaloid 3 with HNO2 in 8%and 21%yields,respectively.Structures of these two compounds were established based on the combination of spectroscopic data,including HRESIMS,1D and 2D NMR data.A plausible mechanism for the formation of 4 and 5 is also presented.

  2. Synthetic studies on neoclerodane diterpenes from Salvia splendens: oxidative modifications of ring A.

    Science.gov (United States)

    Fontana, Gianfranco; Savona, Giuseppe; Rodríguez, Benjamín; Dersch, Christina M; Rothman, Richard B; Prisinzano, Thomas E

    2009-02-21

    Salvinorin A (1), a neoclerodane diterpene from the hallucinogenic mint Salvia divinorum, is the only known naturally occurring non-nitrogenous and specific κ-opioid agonist. Some oxidative modifications of the A ring in the congeners of 1 isolated from Salvia splendens salviarin, splenolide B, splendidin and in the non-natural 8-epi-salviarin gave new derivatives, some of which were tested as agonists at opioid receptors. However, none of these compounds were active. The presence of the C-18, C-19 lactone could be at the origin of the observed lack of binding affinity.

  3. New Non-Toxic Semi-Synthetic Derivatives from Natural Diterpenes Displaying Anti-Tuberculosis Activity.

    Science.gov (United States)

    Matos, Priscilla M; Mahoney, Brian; Chan, Yohan; Day, David P; Cabral, Mirela M W; Martins, Carlos H G; Santos, Raquel A; Bastos, Jairo K; Page, Philip C Bulman; Heleno, Vladimir C G

    2015-10-07

    We report herein the synthesis of six diterpene derivatives, three of which are new, generated through known organic chemistry reactions that allowed structural modification of the existing natural products kaurenoic acid (1) and copalic acid (2). The new compounds were fully characterized using high resolution mass spectrometry, infrared spectroscopy, ¹H- and (13)C-NMR experiments. We also report the evaluation of the anti-tuberculosis potential for all compounds, which showed some promising results for Micobacterium tuberculosis inhibition. Moreover, the toxicity for each of the most active compounds was also assessed.

  4. Synthesis and Sensory Evaluation of ent-Kaurane Diterpene Glycosides

    Directory of Open Access Journals (Sweden)

    Venkata Sai Prakash Chaturvedula

    2012-07-01

    Full Text Available Catalytic hydrogenation of the three ent-kaurane diterpene glycosides isolated from Stevia rebaudiana, namely rubusoside, stevioside, and rebaudioside-A has been carried out using Pd(OH2 and their corresponding dihydro derivatives have been isolated as the products. Synthesis of reduced steviol glycosides was performed using straightforward chemistry and their structures were characterized on the basis of 1D and 2D NMR spectral data and chemical studies. Also, we report herewith the sensory evaluation of all the reduced compounds against their corresponding original steviol glycosides and sucrose for the sweetness property of these molecules.

  5. Antibacterial Activities of a New Brominated Diterpene from Borneon Laurencia spp

    Science.gov (United States)

    Vairappan, Charles Santhanaraju; Ishii, Takahiro; Lee, Tan Kai; Suzuki, Minoru; Zhaoqi, Zhan

    2010-01-01

    In our continuous interest to study the diversity of halogenated metabolites of Malaysian species of the red algal genus Laurencia, we examined the chemical composition of five populations of unrecorded Laurencia sp. A new brominated diterpene, 10-acetoxyangasiol (1), and four other known metabolites, aplysidiol (2), cupalaurenol (3), 1-methyl-2,3,5-tribromoindole (4), and chamigrane epoxide (5), were isolated and identified. Isolated metabolites exhibited potent antibacterial activities against clinical bacteria, Staphylococcus aureus, Staphylococcus sp., Streptococcus pyogenes, Salmonella sp. and Vibrio cholerae. PMID:20631866

  6. Henrin A: A New Anti-HIV Ent-Kaurane Diterpene from Pteris henryi

    Directory of Open Access Journals (Sweden)

    Wan-Fei Li

    2015-11-01

    Full Text Available Henrin A (1, a new ent-kaurane diterpene, was isolated from the leaves of Pteris henryi. The chemical structure was elucidated by analysis of the spectroscopic data including one-dimensional (1D and two-dimensional (2D NMR spectra, and was further confirmed by X-ray crystallographic analysis. The compound was evaluated for its biological activities against a panel of cancer cell lines, dental bacterial biofilm formation, and HIV. It displayed anti-HIV potential with an IC50 value of 9.1 µM (SI = 12.2.

  7. Two New Phorbol-Type Diterpene Esters from Synadenium grantii Hook F. Leaves

    Directory of Open Access Journals (Sweden)

    Emad M. Hassan

    2012-03-01

    Full Text Available Two new phorbol-type diterpene esters has been isolated from the chloroform extract of the leaves of Synadenium grantii Hook F. and identified as 3,4,12,13-tetraacetylphorbol-20-phenylacetate and 4-deoxyphor bol-12,13-ditiglate for which the trivial names Synagrantol A & B respectively, were adopted. Furthermore, two known triterpenes were isolated. The structures of all isolated compounds were established by 1D and 2D NMR spectroscopy including 1H and 13C NMR, HSQC and HMBC techniques. The cytotoxicity as well as the antiparasitic activity of the chloroform extract was performed and proved to be active.

  8. Henrin A: A New Anti-HIV Ent-Kaurane Diterpene from Pteris henryi.

    Science.gov (United States)

    Li, Wan-Fei; Wang, Juan; Zhang, Jing-Jie; Song, Xun; Ku, Chuen-Fai; Zou, Juan; Li, Ji-Xin; Rong, Li-Jun; Pan, Lu-Tai; Zhang, Hong-Jie

    2015-11-24

    Henrin A (1), a new ent-kaurane diterpene, was isolated from the leaves of Pteris henryi. The chemical structure was elucidated by analysis of the spectroscopic data including one-dimensional (1D) and two-dimensional (2D) NMR spectra, and was further confirmed by X-ray crystallographic analysis. The compound was evaluated for its biological activities against a panel of cancer cell lines, dental bacterial biofilm formation, and HIV. It displayed anti-HIV potential with an IC50 value of 9.1 µM (SI = 12.2).

  9. Antibacterial Activities of a New Brominated Diterpene from Borneon Laurencia spp.

    Directory of Open Access Journals (Sweden)

    Charles Santhanaraju Vairappan

    2010-05-01

    Full Text Available In our continuous interest to study the diversity of halogenated metabolites of Malaysian species of the red algal genus Laurencia, we examined the chemical composition of five populations of unrecorded Laurencia sp. A new brominated diterpene, 10-acetoxyangasiol (1, and four other known metabolites, aplysidiol (2, cupalaurenol (3, 1-methyl-2,3,5-tribromoindole (4, and chamigrane epoxide (5, were isolated and identified. Isolated metabolites exhibited potent antibacterial activities against clinical bacteria, Staphylococcus aureus, Staphylococcus sp., Streptococcus pyogenes, Salmonella sp. and Vibrio cholerae.

  10. Chlorabietols A-C, Phloroglucinol-Diterpene Adducts from the Chloranthaceae Plant Chloranthus oldhamii.

    Science.gov (United States)

    Xiong, Juan; Hong, Zhi-Lai; Gao, Li-Xin; Shen, Jie; Liu, Shu-Ting; Yang, Guo-Xun; Li, Jia; Zeng, Huaqiang; Hu, Jin-Feng

    2015-11-01

    Three unprecedented phloroglucinol-diterpene adducts, chlorabietols A-C (1-3), were isolated from the roots of the rare Chloranthaceae plant Chloranthus oldhamii. They represent a new class of compounds, featuring an abietane-type diterpenoid coupled with different alkenyl phloroglucinol units by forming a 2,3-dihydrofuran ring. Their structures were elucidated by detailed spectroscopic analysis, molecular modeling studies, and electronic circular dichroism calculations. Compounds 1-3 showed inhibitory activity against protein tyrosine phosphatase 1B (PTP1B) with IC50 values of 12.6, 5.3, and 4.9 μM, respectively.

  11. 4α-Acetoxyamijidictyol - A New Antifeeding Dolastane Diterpene from the Brazilian Brown Alga Canistrocarpus cervicornis.

    Science.gov (United States)

    Miguel Bianco, Éverson; Martins Francisco, Thiago; Basílio Pinheiro, Carlos; Bagueira de Vasconcellos Azeredo, Rodrigo; Laneuville Teixeira, Valéria; Crespo Pereira, Renato

    2015-11-01

    Chemical investigation of the CH2 Cl2 crude extract from the brown alga Canistrocarpus cervicornis (Dictyotaceae) led to isolation of one new (1) and four previously reported dolastane diterpenes (2-5). Their structures were characterized by 1D- and 2D-NMR spectroscopic techniques, including a full single crystal X-ray diffraction analysis for 1, 2, and 4. In addition, the new structure 1 was assayed as chemical defense inhibiting the feeding by the sea urchin Lytechinus variegatus. This study constitutes an additional report broadening the known spectrum of action and defensive roles of secondary metabolites of the C. cervicornis and Dictyotales species.

  12. Two new ent-kaurane-type diterpene glycosides from zucchini (Cucurbita pepo L.) seeds.

    Science.gov (United States)

    Kikuchi, Takashi; Ando, Hiromi; Maekawa, Ken-Ichiro; Arie, Hiroki; Yamada, Takeshi; Tanaka, Reiko

    2015-12-01

    Two new ent-kaurane diterpene glycosides; 12α-(β-d-glucopyranosyloxy)-7β-hydroxykaurenolide (1) and 7β-(β-d-glucopyranosyloxy)-12α-hydroxykaurenolide (2), a new steroid; (24S)-stigmasta-7,22E,25-trien-3-one (12), and known compounds (3-11, 13-14) were isolated from zucchini (Cucurbita pepo L.) seeds. The absolute structures of 1 and 2 were determined by acid hydrolysis and application of a modified Moscher's method. Furthermore, isolated compounds (1-14), and a derivative, 1a, were evaluated for their inhibitory effects on macrophage activation by an inhibitory assay of nitric oxide (NO) production.

  13. Anti-inflammatory diterpene dimers from the root barks of Aphanamixis grandifolia.

    Science.gov (United States)

    Zhang, Hong-Jian; Zhang, Yang-Mei; Luo, Jian-Guang; Luo, Jun; Kong, Ling-Yi

    2015-07-21

    A total of 14 new diterpene dimers, aphanamenes C-P ()1-14, with four known homologous compounds were isolated from the root barks of Aphanamixis grandifolia Bl. The structures of these compounds were elucidated by spectroscopic analyses, and their absolute configurations were determined using the CD exciton chirality method. In addition, all the compounds exhibited significant inhibition of lipopolysaccharide-induced nitric oxide production in RAW264.7 macrophages, with IC50 values between 7.75 and 38.23 μM.

  14. A comparative study of two clerodane diterpenes from Baccharis trimera (Less.) DC. on the influx and mobilization of intracellular calcium in rat cardiomyocytes.

    Science.gov (United States)

    Garcia, Francisca Adilfa de Oliveira; Tanae, Mirtes Midori; Torres, Luce Maria Brandão; Lapa, Antônio José; de Lima-Landman, Maria Teresa Riggio; Souccar, Caden

    2014-01-01

    Baccharis trimera (Less.) D.C. (Asteraceae) is a medicinal species native to South America and used in Brazilian folk medicine to treat gastrointestinal and liver diseases, kidney disorders and diabetes. The aqueous extract (AE) of the aerial parts of this species presented two mainly constituents: the ent-clerodane diterpene (Fig. 1) and the neo-clerodane diterpene (Fig. 2). The objective of this work was to study their activities on the blockade of Ca(2+)-induced contractions in KCL-depolarized rat portal vein preparations, and on the influx and mobilization of cytosolic calcium in rat cardiomyocytes by fluorescence measurements. The results showed that both the neo- and the ent-clerodane diterpenes reduced the maximal contractions induced by CaCl2, in KCl depolarized rat portal vein preparations, without modifying the EC50. The data on the concentration of cytosolic calcium ([Ca(2+)]c) showed that, while the neo-clerodane diterpene stimulates the mobilization of [Ca(2+)]c in rat cardiomyocytes, this effect was not observed with the ent-clerodane diterpene. On the other hand, the influx of calcium was not altered by the neo-clerodane diterpene, but was reduced in the presence of the ent-clerodane diterpene, indicating that this compound induces a blockade of the voltage-dependent calcium channels.

  15. Anti-malarial, anti-algal, anti-tubercular, anti-bacterial, anti-photosynthetic, and anti-fouling activity of diterpene and diterpene isonitriles from the tropical marine sponge Cymbastela hooperi.

    Science.gov (United States)

    Wright, Anthony D; McCluskey, Adam; Robertson, Mark J; MacGregor, Kylie A; Gordon, Christopher P; Guenther, Jana

    2011-01-21

    In an investigation into their potential ecological role(s), a group of mainly diterpene isonitriles, nine in total, isolated from the tropical marine sponge Cymbastela hooperi, and the sesquiterpene axisonitrile-3, isolated from the tropical marine sponge Acanthella kletra, were evaluated in a series of bioassays including anti-fouling, anti-algal, anti-photosynthetic, anti-bacterial (Gram +ve and -ve), anti-fungal, and anti-tubercular. The results of these assays showed that all of the tested compounds, with the exception of diterpene 9, were active in at least two of the applied test systems, with axisonitrile-3 (10) and diterpene isonitrile 1 being the two most active compounds overall, closely followed by diterpene isonitrile 3. Based on the results of the photosynthetic study a molecular modelling investigation was undertaken with all of the compounds used in that study. The results showed a positive correlation between reduction in photosynthetic activity and the interaction of the modelled compounds with a potential enzyme active site.

  16. Characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (Rosmarinus officinalis) glandular trichomes

    DEFF Research Database (Denmark)

    Brückner, Kathleen; Božić, Dragana; Manzano, David

    2014-01-01

    normal CDP to produce an abietane diterpene. Comparison to the already characterized diterpene synthase from Salvia miltiorrhiza (SmKSL) demonstrates that the product of RoKSL1 and RoKSL2 is miltiradiene. Expression analysis supports a major contributing role for RoKSL2. Like SmKSL and the sclareol...... synthase from Salvia sclarea, RoKSL1/2 are diterpene synthases of the TPS-e group which have lost the internal gamma-domain. Furthermore, phylogenetic analysis indicates that RoKSL1 and RoKSL2 belong to a distinct group of KSL enzymes involved in specialized metabolism which most likely emerged before...

  17. Chemotaxonomy of three genera of the Annonaceae family using self-organizing maps and {sup 13}C NMR data of diterpenes

    Energy Technology Data Exchange (ETDEWEB)

    Scotti, Luciana; Tavares, Josean Fechine; Silva, Marcelo Sobral da [Universidade Federal da Paraiba (UFPB), Joao Pessoa, PB (Brazil). Dept. de Ciencias Farmaceuticas; Falcao, Emanuela Viana; Silva, Luana de Morais e; Soares, Gabriela Cristina da Silva; Scotti, Marcus Tullius, E-mail: mtscotti@ccae.ufpb.br [Universidade Federal da Paraiba (UFPB), Rio Tinto, PB (Brazil). Dept. de Engenharia e Meio Ambiente

    2012-07-01

    The Annonaceae family is distributed throughout Neotropical regions of the world. In Brazil, it covers nearly all natural formations particularly Annona, Xylopia and Polyalthia and is characterized chemically by the production of sources of terpenoids (mainly diterpenes), alkaloids, steroids, polyphenols and, flavonoids. Studies from {sup 13}C NMR data of diterpenes related with their botanical occurrence were used to generate self-organizing maps. Results corroborate those in the literature obtained from morphological and molecular data for three genera and the model can be used to project other diterpenes. Therefore, the model produced can predict which genera are likely to contain a compound. (author)

  18. Coffee diterpenes prevent benzo[a]pyrene genotoxicity in rat and human culture systems.

    Science.gov (United States)

    Cavin, C; Bezencon, C; Guignard, G; Schilter, B

    2003-06-27

    The coffee-specific diterpenes cafestol and kahweol (C+K) have been identified as two important chemoprotective agents in coffee. In the present study, the potential preventive effects of C+K against the genotoxicity of B[a]P were investigated in rat primary hepatocytes and in human bronchial Beas-2B cells. Several independent mechanisms were identified and their respective contribution to the overall protective effects was determined. A marked dose-dependent inhibition by C+K of B[a]P DNA-binding was found in cells of both origins. However, data showed that the significant induction by C+K of the detoxifying enzyme GST-Yp subunit is the key mechanism of protection against B[a]P DNA-binding in rat liver. In contrast, the phase I-mediated mechanism where C+K produce an inhibition of CYP 1A1 induction by B[a]P is of key significance for the C+K protection in human Beas-2B cells. Moreover, this effect suggests a novel mechanism of chemoprotection by the coffee diterpenes cafestol and kahweol.

  19. A new family of synthetic diterpenes that regulates cytokine synthesis by inhibiting IkappaBalpha phosphorylation.

    Science.gov (United States)

    Chao, Ta-Hsiang; Lam, Thanh; Vong, Binh G; Través, Paqui G; Hortelano, Sonsoles; Chowdhury, Chinmay; Bahjat, F Rena; Lloyd, G Kenneth; Moldawer, Lyle L; Boscá, Lisardo; Palladino, Michael A; Theodorakis, Emmanuel A

    2005-01-01

    The synthesis and the biological evaluation of a new family diterpenes are presented. The synthetic studies were inspired by the structural framework of acanthoic acid (1) and yielded a family of compounds that were evaluated as anti-inflammatory agents. Among them, compounds 2, 10, 12, and 16 exhibited a very low nonspecific cytotoxicity and inhibited the synthesis of TNF-alpha with greater than 65 % efficacy at low micromolar concentrations. Cytokine-specificity studies revealed that these compounds also inhibited the synthesis of the proinflammatory cytokines IL-1beta and IL-6, while inhibition of IL-1ra and IL-8 synthesis was marginal and only occurred at high concentrations. Further studies, through EMSA and Western blot analyses, indicated that these compounds decreased the extent of phosphorylation of IkappaBalpha; this suggests that they exert their anti-inflammatory profile by inhibiting NF-kappaB-mediated cytokine synthesis. These findings imply that these diterpenes represent promising leads for the development of novel anti-inflammatory agents.

  20. Discovery and biological evaluation of the novel naturally occurring diterpene pepluanone as antiinflammatory agent.

    Science.gov (United States)

    Corea, Gabriella; Fattorusso, Ernesto; Lanzotti, Virginia; Di Meglio, Paola; Maffia, Pasquale; Grassia, Gianluca; Ialenti, Armando; Ianaro, Angela

    2005-11-03

    From the whole plant of Euphorbia peplus L., a new diterpene based on a rare pepluane skeleton, named pepluanone (1), was isolated together with a known pepluane diterpene (2). The stereostructure of pepluanone was determined on the basis of an extensive NMR study, MS data, and chemical reaction. The ability of these compounds to act as antiinflammatory agents has been evaluated for the first time by in vivo tests on carrageenin-induced rat paw edema, an experimental model of acute inflammation. Comparison of the bioactivity of pepluanone and compound 2 in terms of chemical structure, evidenced the high efficiency of pepluanone and the absence of appreciable activity for compound 2, thus giving a first insight into the structure-activity relationship. Further in vitro experiments performed on pepluanone let us hypothesize that its activity could be explained in reducing the production of nitric oxide, prostaglandin E(2), and TNF-alpha by inhibiting the expression of inducible nitric oxide synthase, cyclooxygenase-2, and TNF-alpha mRNA through the down-regulation of NF-kappaB binding activity.

  1. Semi-synthesis of oxygenated dolabellane diterpenes with highly in vitro anti-HIV-1 activity.

    Science.gov (United States)

    Pardo-Vargas, Alonso; Ramos, Freddy A; Cirne-Santos, Claudio Cesar; Stephens, Paulo Roberto; Paixão, Izabel Christina Palmer; Teixeira, Valeria Laneuville; Castellanos, Leonardo

    2014-09-15

    Research on dolabellane diterpenes of brown algae Dictyota spp. has shown that these diterpenoids have strong anti-HIV-1 activity, but there are not data about antiviral activity of dolabellane diterpenes isolated from octocorals, which are antipodes of those isolated from the brown algae. Dolabellanes 13-keto-1(R),11(S)-dolabella-3(E),7(E),12(18)-triene (1) and β-Araneosene (2) were isolated from the Caribbean octocoral Eunicea laciniata, and both showed low anti-HIV-1 activity and low toxicity. Since it was shown that oxygenated dolabellanes from algae have better anti-HIV-1 activity, in this work some derivatives of the main dolabellane of E. laciniata1 were obtained by epoxidation (3), epoxide opening (4), and allylic oxidation (5). The derivatives showed significant improvement in the anti-HIV-1potency (100-fold), being compounds 3 and 5 the most active ones. Their high antiviral activities, along with their low cytotoxicity, make them promissory antiviral compounds; and it is worth noting that the absolute configuration at the ring junction in the dolabellane skeleton does not seem to be determinant in the antiviral potency of these diterpeneoids.

  2. Marine Diterpenes: Molecular Modeling of Thrombin Inhibitors with Potential Biotechnological Application as an Antithrombotic

    Science.gov (United States)

    Pereira, Rebeca Cristina Costa; Lourenço, André Luiz; Terra, Luciana; Abreu, Paula Alvarez; Laneuville Teixeira, Valéria; Castro, Helena Carla

    2017-01-01

    Thrombosis related diseases are among the main causes of death and incapacity in the world. Despite the existence of antithrombotic agents available for therapy, they still present adverse effects like hemorrhagic risks which justify the search for new options. Recently, pachydictyol A, isopachydictyol A, and dichotomanol, three diterpenes isolated from Brazilian marine brown alga Dictyota menstrualis were identified as potent antithrombotic molecules through inhibition of thrombin, a key enzyme of coagulation cascade and a platelet agonist. Due to the biotechnological potential of these marine metabolites, in this work we evaluated their binding mode to thrombin in silico and identified structural features related to the activity in order to characterize their molecular mechanism. According to our theoretical studies including structure-activity relationship and molecular docking analysis, the highest dipole moment, polar surface area, and lowest electronic density of dichotomanol are probably involved in its higher inhibition percentage towards thrombin catalytic activity compared to pachydictyol A and isopachydictyol A. Interestingly, the molecular docking studies also revealed a good shape complementarity of pachydictyol A and isopachydictyol A and interactions with important residues and regions (e.g., H57, S195, W215, G216, and loop-60), which probably justify their thrombin inhibitor effects demonstrated in vitro. Finally, this study explored the structural features and binding mode of these three diterpenes in thrombin which reinforced their potential to be further explored and may help in the design of new antithrombotic agents. PMID:28335516

  3. Marine Diterpenes: Molecular Modeling of Thrombin Inhibitors with Potential Biotechnological Application as an Antithrombotic

    Directory of Open Access Journals (Sweden)

    Rebeca Cristina Costa Pereira

    2017-03-01

    Full Text Available Thrombosis related diseases are among the main causes of death and incapacity in the world. Despite the existence of antithrombotic agents available for therapy, they still present adverse effects like hemorrhagic risks which justify the search for new options. Recently, pachydictyol A, isopachydictyol A, and dichotomanol, three diterpenes isolated from Brazilian marine brown alga Dictyota menstrualis were identified as potent antithrombotic molecules through inhibition of thrombin, a key enzyme of coagulation cascade and a platelet agonist. Due to the biotechnological potential of these marine metabolites, in this work we evaluated their binding mode to thrombin in silico and identified structural features related to the activity in order to characterize their molecular mechanism. According to our theoretical studies including structure-activity relationship and molecular docking analysis, the highest dipole moment, polar surface area, and lowest electronic density of dichotomanol are probably involved in its higher inhibition percentage towards thrombin catalytic activity compared to pachydictyol A and isopachydictyol A. Interestingly, the molecular docking studies also revealed a good shape complementarity of pachydictyol A and isopachydictyol A and interactions with important residues and regions (e.g., H57, S195, W215, G216, and loop-60, which probably justify their thrombin inhibitor effects demonstrated in vitro. Finally, this study explored the structural features and binding mode of these three diterpenes in thrombin which reinforced their potential to be further explored and may help in the design of new antithrombotic agents.

  4. Studies on Antibacterial QSAR of Diterpene Quinones from Sclvic Przewalskii Maxim

    Institute of Scientific and Technical Information of China (English)

    XUE Ming; SHI Yan-bin; CUI Yin; ZHOU Zong-tian; ZHANG Bin; LUO Yong-jiang; XIA Wen-jiang; ZHAO Rong-cai; WANG Han-qing

    2001-01-01

    The antibacierial (staphylococcus aureus) activities of 23 diterpene quinones from the Salvia przewalskii Maxim, the artificial synthesis ones and that of the metabolism in pigs were examined in an effort to study the quantitative structure activity relationship with the Free-Wilson method. With the application of the multipiy regression, the Free-Wilson pattern was established by the dummy (indicator) variable of the structural fragments or substituents. The results showed: (1) The diketone moiety played the fundamental role in antibacterial activities, ortho-quinones was higher active than para-quinones. (2) The single bond existed between the C15 and the C16 at D-ring led to higher activity. (3) The saturation at the A-ring led to higher activity. (4) The components adjacent to nitrogen substituents at C15 led to higher activity. (5)The hydroxylation or dehydrogenation at A-ring led to less activity. The research results can provide the theoretical basis for developing a new antibacterial drug from the Salvia diterpene quinones.

  5. Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase.

    Science.gov (United States)

    Chen, Mengbin; Chou, Wayne K W; Toyomasu, Tomonobu; Cane, David E; Christianson, David W

    2016-04-15

    Fusicoccin A is a diterpene glucoside phytotoxin generated by the fungal pathogen Phomopsis amygdali that causes the plant disease constriction canker, first discovered in New Jersey peach orchards in the 1930s. Fusicoccin A is also an emerging new lead in cancer chemotherapy. The hydrocarbon precursor of fusicoccin A is the tricyclic diterpene fusicoccadiene, which is generated by a bifunctional terpenoid synthase. Here, we report X-ray crystal structures of the individual catalytic domains of fusicoccadiene synthase: the C-terminal domain is a chain elongation enzyme that generates geranylgeranyl diphosphate, and the N-terminal domain catalyzes the cyclization of geranylgeranyl diphosphate to form fusicoccadiene. Crystal structures of each domain complexed with bisphosphonate substrate analogues suggest that three metal ions and three positively charged amino acid side chains trigger substrate ionization in each active site. While in vitro incubations reveal that the cyclase domain can utilize farnesyl diphosphate and geranyl diphosphate as surrogate substrates, these shorter isoprenoid diphosphates are mainly converted into acyclic alcohol or hydrocarbon products. Gel filtration chromatography and analytical ultracentrifugation experiments indicate that full-length fusicoccadiene synthase adopts hexameric quaternary structure, and small-angle X-ray scattering data yield a well-defined molecular envelope illustrating a plausible model for hexamer assembly.

  6. 1H and 13C NMR signal assignments of a novel Baeyer-Villiger originated diterpene lactone.

    Science.gov (United States)

    Vieira, Henriete S; Takahashi, Jacqueline A; Gunatilaka, A A Leslie; Boaventura, Maria Amélia D

    2006-02-01

    A highly rearranged novel dilactone was the single product isolated from Baeyer-Villiger oxidation of a norketone prepared from grandiflorenic acid, a natural kaurane diterpene. The complete 1H and 13C NMR assignment is presented for this novel compound that showed discrete in vitro antibacterial activity. Copyright 2005 John Wiley & Sons, Ltd.

  7. Health effects of unfiltered coffee - diterpenes in coffee and their effects on blood lipids and liver enzymes in man.

    NARCIS (Netherlands)

    Urgert, R.

    1997-01-01

    Boiled coffee raises blood levels of cholesterol and triglycerides in man. Cafestol and kahweol are responsible. These diterpenes also raise blood levels of alanine aminotransferase (ALT). The objective of the present studies was to further specify the health effects of cafestol and kahweol.Unfilter

  8. Vibrational Circular Dichroism Absolute Configuration of 9,12-Cyclomulin-13-ol, a Diterpene from Azorella and Laretia Species.

    Science.gov (United States)

    Muñoz, Marcelo A; San-Martín, Aurelio; Joseph-Nathan, Pedro

    2015-08-01

    The absolute configuration of the diterpenoid 9,12-cyclomulin-13-ol (1), a constituent of Azorella and Laretia species, has been established by vibrational circular dichroism spectroscopy in combination with density functional theory calculations. The obtained normal diterpene absolute configuration confirms that of azorellanol (2), which was determined by single crystal X-ray diffraction.

  9. Mechanistic studies on the lipid-raising coffee diterpenes cafestol and kahweol in monkeys, mice and man

    NARCIS (Netherlands)

    Roos, de B.

    2000-01-01

    Cafestol and kahweol are lipid-raising diterpenes present in unfiltered coffee. The objective of this thesis was to study their lipid-raising action in man. Unravelling this action might lead to new insights into the regulation of serum cholesterol levels.We first studied the absorption and urinary

  10. The Primary Diterpene Synthase Products of Picea abies Levopimaradiene/Abietadiene Synthase (PaLAS) Are Epimers of a Thermally Unstable Diterpenol*

    Science.gov (United States)

    Keeling, Christopher I.; Madilao, Lina L.; Zerbe, Philipp; Dullat, Harpreet K.; Bohlmann, Jörg

    2011-01-01

    The levopimaradiene/abietadiene synthase from Norway spruce (Picea abies; PaLAS) has previously been reported to produce a mixture of four diterpene hydrocarbons when incubated with geranylgeranyl diphosphate as the substrate: levopimaradiene, abietadiene, neoabietadiene, and palustradiene. However, variability in the assay products observed by GC-MS of this and orthologous conifer diterpene synthases over the past 15 years suggested that these diterpenes may not be the initial enzyme assay products but are rather the products of dehydration of an unstable alcohol. We have identified epimers of the thermally unstable allylic tertiary alcohol 13-hydroxy-8(14)-abietene as the products of PaLAS. The identification of these compounds, not previously described in conifers, as the initial products of PaLAS has considerable implications for our understanding of the complexity of the biosynthetic pathway of the structurally diverse diterpene resin acids of conifer defense. PMID:21518766

  11. Identification of UV-Induced Diterpenes Including a New Diterpene Phytoalexin, Phytocassane F, from Rice Leaves by Complementary GC/MS and LC/MS Approaches.

    Science.gov (United States)

    Horie, Kiyotaka; Inoue, Yasuno; Sakai, Miki; Yao, Qun; Tanimoto, Yosuke; Koga, Jinichiro; Toshima, Hiroaki; Hasegawa, Morifumi

    2015-04-29

    Rice phytoalexins are regarded as one of the most important weapons against pathogenic microorganisms. We attempted to identify novel phytoalexins and their derivatives using GC/MS and LC/MS analyses. Diterpene derivatives, 9β-pimara-7,15-diene-3β,6β,19-triol, 1, stemar-13-en-2α-ol, 2, and 1α,2α-dihydroxy-ent-12,15-cassadiene-3,11-dione, 3, were isolated from UV-irradiated rice leaves by chromatographic methods. These structures were confirmed by 1D- and 2D-NMR and MS analyses. Interestingly, all three compounds were accumulated following an infection by the rice blast pathogen Magnaporthe oryzae. Compounds 1 and 2 exhibited weak antifungal activity and may be the biosynthetic intermediates of rice phytoalexins momilactones and oryzalexin S, respectively. Compound 3 exhibited relatively high inhibitory activity against the fungal mycelial growth of M. oryzae to the same extent as the known phytoalexin phytocassane A. We conclude that 3 is a member of the cassane-type phytoalexin family and propose the name phytocassane F.

  12. In Vitro Antimicrobial Activity of Plant-Derived Diterpenes against Bovine Mastitis Bacteria

    Directory of Open Access Journals (Sweden)

    Rodrigo C. S. Veneziani

    2013-07-01

    Full Text Available We evaluated the antibacterial activity of three diterpenes isolated from natural sources against a panel of microorganisms responsible for bovine mastitis. ent-Copalic acid (CA was the most active metabolite, with promising MIC values (from 1.56 to 6.25 µg mL−1 against Staphylococcus aureus (ATCC and clinical isolate, Staphylococcus epidermidis, Streptococcus agalactiae, and Streptococcus dysgalactiae. We conducted time-kill assays of CA against S. aureus, a commensal organism considered to be a ubiquitous etiological agent of bovine mastitis in dairy farms worldwide. In the first 12 h, CA only inhibited the growth of the inoculums (bacteriostatic effect, but its bactericidal effect was clearly noted thereafter (between 12 and 24 h. In conclusion, CA should be considered for the control of several Gram-positive bacteria related to bovine mastitis.

  13. New C₂₀-gibberellin diterpene from the leaves of Schefflera sessiliflora De P. V.

    Science.gov (United States)

    Nguyen, Tan Phat; Tran, Thi Thao Vy; Mai, Dinh Tri; Le, Tien Dung; Phan, Nhat Minh; Bui, Trong Dat

    2015-01-01

    From the leaves of Schefflera sessiliflora De P. V., one new C20-gibberellin diterpene 2β,12β-dihydroxygibberellin (12β-hydroxy-GA110 or 2β-hydroxy-GA112) (1), together with three known compounds, trans-tiliroside (2), kaempferol 3-O-β-D-glucuronopyranoside (3), 5-p-trans-coumaroylquinic acid (4), was isolated for the first time from the genus Schefflera by various chromatography methods. Their structures were elucidated by IR, UV, HR-ESI-MS, NMR 1D and 2D experiments and comparison with previous reported data. The α-glucosidase inhibitory activity of all compounds was measured. The isolates (2, 3) showed better α-glucosidase inhibitory activity (IC50 = 134.60, 147.10 μM, respectively) than the standard drug acarbose (IC50 = 214.50 μM).

  14. Isolation and structural characterization of a new minor diterpene glycoside from Stevia rebaudiana.

    Science.gov (United States)

    Chaturvedula, Venkata Sai Prakash; Zamora, Julian

    2014-12-01

    From the commercial extract of the leaves of the sweet plant Stevia rebaudiana Bertoni obtained from Sinochem Qingdao Co. Ltd., a new diterpene glycoside having three β-D-glucopyranosyl units of which two of them were connected in a relatively rare linkage of 3-β-D-glucobiosyl substitution at C-19 position of the aglycone steviol. The structure of the new compound has been characterized as 13-β-D-glucopyranosyloxy ent-kaur-16-en-19-oic acid-[(3-O-β-D- glucopyranosyl-β-D-glucopyranosyl) ester (1) on the basis of extensive 1D (1H and 13C) and 2D NMR (TOCSY, HMQC, and HMBC), and High Resolution mass spectroscopic data as well as hydrolysis studies.

  15. The synthesis and antibacterial activity of pyrazole-fused tricyclic diterpene derivatives.

    Science.gov (United States)

    Yu, Li-Gang; Ni, Teng-Feng; Gao, Wei; He, Yuan; Wang, Ying-Ying; Cui, Hai-Wei; Yang, Cai-Guang; Qiu, Wen-Wei

    2015-01-27

    The diterpenoid compound 5 was identified as an antibacterial lead in our screening of small synthetic natural product-like (NPL) library. A series of novel diterpene derivatives were synthesized and investigated for their activity against Staphylococcus aureus Newman strain and multidrug-resistant strains (NRS-1, NRS-70, NRS-100, NRS-108 and NRS-271). Among the compounds tested, 42 and 43 showed highest activity with a MIC of 1 μg/mL against strain Newman, 45 and 52 showed the most potent activity with MIC values of 0.71-3.12 μg/mL against five multidrug-resistant S. aureus. All high-antimicrobial active compounds showed no obvious toxicity to human fibroblast (HAF) cells at the MIC concentration.

  16. Quesnoin, a novel pentacyclic ent-diterpene from 55 million years Old Oise Amber.

    Science.gov (United States)

    Jossang, Jean; Bel-Kassaoui, Hakima; Jossang, Akino; Seuleiman, Mannan; Nel, André

    2008-01-18

    Amber, fossilized tree resin, found at the Oise River area of the Paris basin (France) was dated as being 55 million years old. Quesnoin, a novel unique pure organic compound, was isolated from Oise amber. 1H and 13C NMR spectroscopic analysis indicated an unknown diterpene skeleton, quesnane. The absolute configurations of the eight chiral centers of quesnoin were determined to be 4S, 5S, 8R, 9S, 10S, 13S, 14R, and 16S by chiral auxiliary (R)- and (S)-phenylglycine methyl ester derivatization. Quesnoin allowed us to disclose the tree producer, corresponding to modern Hymenaea oblongifolia, Fabaceae, a subfamily of Caesalpiniaceae, one of the oldest angiosperm. The presence of the Amazon rainforest tree, H. oblongifolia, indicated that the climate of the Paris basin might have been tropical in the early Eocene period, 55 million years ago.

  17. New Ent-Kaurane-Type Diterpene Glycosides and Benzophenone from Ranunculus muricatus Linn.

    Science.gov (United States)

    Wu, Bi-Ling; Zou, Hui-Liang; Qin, Fang-Min; Li, Hong-Yu; Zhou, Guang-Xiong

    2015-12-15

    Two new ent-kaurane diterpene glycosides, ranunculosides A (1) and B (2), and a new benzophenone, ranunculone C (3), were isolated from the aerial part of Ranunculus muricatus Linn. The chemical structures of compounds 1-3 were established to be (2S)-ent-kauran-2β-ol-15-en-14-O-β-d-glucopyranoside, (2S,4S)-ent-kauran-2β,18-diol-15-en-14-O-β-d-glucopyranoside, and (R)-3-[2-(3,4-dihydroxybenzoyl)-4,5-dihydroxy-phenyl]-2-hydroxylpropanoic acid, respectively, by spectroscopic data and chemical methods. The absolute configuration of 1 was determined by the combinational application of RP-HPLC analysis and Mosher's method.

  18. The natural diterpene tonantzitlolone A and its synthetic enantiomer inhibit cell proliferation and kinesin-5 function.

    Science.gov (United States)

    Pfeffer, Tobias J; Sasse, Florenz; Schmidt, Christoph F; Lakämper, Stefan; Kirschning, Andreas; Scholz, Tim

    2016-04-13

    Tonantzitlolone A, a diterpene isolated from the Mexican plant Stillingia sanguinolenta, shows cytostatic activity. Both the natural product tonantzitlolone A and its synthetic enantiomer induce monoastral spindle formation in cell experiments which indicates inhibitory activity on kinesin-5 mitotic motor molecules. These inhibitory effects on kinesin-5 could be verified in in vitro single-molecule motility assays, where both tonantzitlolones interfered with kinesin-5 binding to its cellular interaction partner microtubules in a concentration-dependent manner, yet with a larger effect of the synthetic enantiomer. In contrast to kinesin-5 inhibition, both tonantzitlolone A enantiomers did not affect conventional kinesin-1 function; hence tonantzitlolones are not unspecific kinesin inhibitors. The observed stronger inhibitory effect of the synthetic enantiomer demonstrates the possibility to enhance the overall moderate anti-proliferative effect of the lead compound tonantzitlolon A by chemical modification.

  19. Antiangiogenic properties of cafestol, a coffee diterpene, in human umbilical vein endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Shuaiyu [Food Biotechnology, University of Science and Technology, 113 Gwahangno, Yuseong-gu, Daejeon 305-333 (Korea, Republic of); Korea Food Research Institute, 516 Baekhyun-dong, Bundang-gu, Songnam, Kyungki-do 463-746 (Korea, Republic of); Yoon, Yeo Cho; Sung, Mi-Jeong; Hur, Haeng-Jeon [Korea Food Research Institute, 516 Baekhyun-dong, Bundang-gu, Songnam, Kyungki-do 463-746 (Korea, Republic of); Park, Jae-Ho, E-mail: jaehoparkmail@gmail.com [Korea Food Research Institute, 516 Baekhyun-dong, Bundang-gu, Songnam, Kyungki-do 463-746 (Korea, Republic of)

    2012-05-11

    Highlights: Black-Right-Pointing-Pointer Cafestol inhibits tube formation and migration of VEGF-stimulated HUVEC. Black-Right-Pointing-Pointer Cafestol inhibits phosphorylation of FAK and Akt. Black-Right-Pointing-Pointer Cafestol decreases NO production. -- Abstract: As angiogenesis plays important roles in tumor growth and metastasis, searching for antiangiogenic compounds is a promising tactic for treating cancers. Cafestol, a diterpene found mainly in unfiltered coffee, provides benefit through varied biological activity, including antitumorigenic, antioxidative, and anti-inflammatory effects. This study aimed to investigate the effects of cafestol on angiogenesis and to uncover the associated mechanism. We show that cafestol inhibits angiogenesis of human umbilical vascular endothelial cells. This inhibition affects the following specific steps of the angiogenic process: proliferation, migration, and tube formation. The inhibitory effects of cafestol are accompanied by decreasing phosphorylation of FAK and Akt and by a decrease in nitric oxide production. Overall, cafestol inhibits angiogenesis by affecting the angiogenic signaling pathway.

  20. A New Abietene Diterpene and Other Constituents from Kaempferia angustifolia Rosc.

    Directory of Open Access Journals (Sweden)

    Abdul Manaf Ali

    2011-04-01

    Full Text Available A new abietene diterpene, kaempfolienol (5S,6S,7S,9S,10S,11R,13S-abiet-8(14-enepenta-6,7,9,11,13-ol, 1, was isolated from a rhizome extract of Kaempferia angustifolia Rosc. along with the known compounds crotepoxide, boesenboxide, zeylenol, 2′-hydroxy-4,4′,6′-trimethoxychalcone, (24S-24-methyl-5α-lanosta-9(11,25-dien-3β-ol, β-sitosterol and β-sitosterol-3-O-β-D-glucopyranoside. The structures of all compounds were elucidated on the basis of mass spectroscopic and NMR data. Zeylenol (2, the major constituent of the plant, was derivatized into diacetate, triacetate and epoxide derivatives through standard organic reactions. The cytotoxic activity of compounds 1, 2 and the zeylenol derivatives was evaluated against the HL-60, MCF-7, HT-29 and HeLa cell lines.

  1. New Ent-Kaurane-Type Diterpene Glycosides and Benzophenone from Ranunculus muricatus Linn.

    Directory of Open Access Journals (Sweden)

    Bi-Ling Wu

    2015-12-01

    Full Text Available Two new ent-kaurane diterpene glycosides, ranunculosides A (1 and B (2, and a new benzophenone, ranunculone C (3, were isolated from the aerial part of Ranunculus muricatus Linn. The chemical structures of compounds 1–3 were established to be (2S-ent-kauran-2β-ol-15-en-14-O-β-d-glucopyranoside, (2S,4S-ent-kauran-2β,18-diol-15-en-14-O-β-d-glucopyranoside, and (R-3-[2-(3,4-dihydroxybenzoyl-4,5-dihydroxy-phenyl]-2-hydroxylpropanoic acid, respectively, by spectroscopic data and chemical methods. The absolute configuration of 1 was determined by the combinational application of RP-HPLC analysis and Mosher’s method.

  2. Antifungal Diterpene Alkaloids from the Caribbean Sponge Agelas citrina: Unified Configurational Assignments of Agelasidines and Agelasines

    Science.gov (United States)

    Stout, E. Paige; Yu, Lily C.

    2013-01-01

    Three new diterpene alkaloids – the hypotaurocyamines, (−)-agelasidines E and F (5–6), and the adeninium salt, agelasine N (9) – were isolated from the Caribbean sponge Agelas citrina along with six known natural products agelasines B–E (7, 10–12), 2-oxo-agelasine B (8), and (−)-agelasidine C (3). The chemical structures of 5, 6 and 9 were elucidated by analysis of NMR spectra and mass spectrometry. This represents the first report of natural products from the sponge A. citrina. Unified assignment of absolute configurations of the new compounds and known compounds were achieved by chemical correlation, quantitative measurements of molar rotations, and comparative analysis by van’t Hoff’s principle of optical superposition. (−)-Agelasidine C (3) exhibited potent antifungal and modest cytotoxic activity against human chronic lymphocytic leukemia (CLL) cells. PMID:24653665

  3. Discovery of Unclustered Fungal Indole Diterpene Biosynthetic Pathways through Combinatorial Pathway Reassembly in Engineered Yeast.

    Science.gov (United States)

    Tang, Man-Cheng; Lin, Hsiao-Ching; Li, Dehai; Zou, Yi; Li, Jian; Xu, Wei; Cacho, Ralph A; Hillenmeyer, Maureen E; Garg, Neil K; Tang, Yi

    2015-11-01

    The structural diversity and biological activities of fungal indole diterpenes (IDTs) are generated in large part by the IDT cyclases (IDTCs). Identifying different IDTCs from IDT biosynthetic pathways is therefore important toward understanding how these enzymes introduce chemical diversity from a common linear precursor. However, IDTCs involved in the cyclization of the well-known aflavinine subgroup of IDTs have not been discovered. Here, using Saccharomyces cerevisiae as a heterologous host and a phylogenetically guided enzyme mining approach, we combinatorially assembled IDT biosynthetic pathways using IDTCs homologues identified from different fungal hosts. We identified the genetically standalone IDTCs involved in the cyclization of aflavinine and anominine and produced new IDTs not previously isolated. The cyclization mechanisms of the new IDTCs were proposed based on the yeast reconstitution results. Our studies demonstrate heterologous pathway assembly is a useful tool in the reconstitution of unclustered biosynthetic pathways.

  4. Insights into Diterpene Cyclization from Structure of Bifunctional Abietadiene Synthase from Abies grandis

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Ke; Gao, Yang; Hoy, Julie A.; Mann, Francis M.; Honzatko, Richard B.; Peters, Reuben J. (Iowa State)

    2013-09-24

    Abietadiene synthase from Abies grandis (AgAS) is a model system for diterpene synthase activity, catalyzing class I (ionization-initiated) and class II (protonation-initiated) cyclization reactions. Reported here is the crystal structure of AgAS at 2.3 {angstrom} resolution and molecular dynamics simulations of that structure with and without active site ligands. AgAS has three domains ({alpha}, {beta}, and {gamma}). The class I active site is within the C-terminal {alpha} domain, and the class II active site is between the N-terminal {gamma} and {beta} domains. The domain organization resembles that of monofunctional diterpene synthases and is consistent with proposed evolutionary origins of terpene synthases. Molecular dynamics simulations were carried out to determine the effect of substrate binding on enzymatic structure. Although such studies of the class I active site do lead to an enclosed substrate-Mg{sup 2+} complex similar to that observed in crystal structures of related plant enzymes, it does not enforce a single substrate conformation consistent with the known product stereochemistry. Simulations of the class II active site were more informative, with observation of a well ordered external loop migration. This 'loop-in' conformation not only limits solvent access but also greatly increases the number of conformational states accessible to the substrate while destabilizing the nonproductive substrate conformation present in the 'loop-out' conformation. Moreover, these conformational changes at the class II active site drive the substrate toward the proposed transition state. Docked substrate complexes were further assessed with regard to the effects of site-directed mutations on class I and II activities.

  5. Use of dietary rosemary diterpenes to inhibit rancid volatiles in lamb meat packed under protective atmosphere.

    Science.gov (United States)

    Ortuño, J; Serrano, R; Bañón, S

    2016-08-01

    The objective of the present study was to determine the inhibitory effect of dietary rosemary diterpenes on the formation of the volatile organic compounds (VOCs) responsible for rancid flavour in raw lamb meat. The lamb diet was supplemented during the fattening stage with two levels (200 and 400 mg/kg feed) of a dietary rosemary extract (DRE) containing carnosic acid and carnosol (1 : 1, w/w). The formation of VOCs (determined by headspace solid-phase microextraction at 40°C and MS) and odour deterioration (assessed by quantitative descriptive analysis) were monitored in meat fillets (longissimus dorsi-lumborum muscle) packed in a 70/30 O2/CO2 protective atmosphere and kept at 2°C for up to 14 days. The raw meat odour deteriorated under pro-oxidizing conditions due to the development of an incipient rancidity caused by the formation of volatiles from lipid oxidation. A total of 46 volatile compounds were determined in lamb headspace: 18 aldehydes, seven alcohols, seven organic acids, six ketones, four furan compounds, two benzene compounds, one ester and one terpenoid. The use of DRE contributed to inhibit VOC formation and rancidity. Heptanal, octanal, nonanal and 2-pentyl-furan were the only VOCs affected (P0.75; Plamb meat. Principal component analysis confirmed that the differences in the VOC profile make it possible to identify whether or not samples have been reinforced with dietary rosemary diterpenes. Thus, VOC profiling can be regarded as a useful tool for assessing the dietary treatments used in sheep to improve the oxidative stability of lamb meat.

  6. Pyrrole Derivatives and Diterpene Alkaloids from the South China Sea Sponge Agelas nakamurai.

    Science.gov (United States)

    Chu, Mei-Jun; Tang, Xu-Li; Qin, Guo-Fei; Sun, Yan-Ting; Li, Lei; de Voogd, Nicole J; Li, Ping-Lin; Li, Guo-Qiang

    2017-07-01

    Two pairs of new non-brominated racematic pyrrole derivatives, (±)-nakamurine D (1) and (±)-nakamurine E (2), two new diterpene alkaloids, isoagelasine C (16) and isoagelasidine B (21), together with 13 known pyrrole derivatives ((±)-3 - 15), five known diterpene alkaloids (17 - 20, 22) were isolated from the South China Sea sponge Agelas nakamurai. The racemic mixtures, compounds 1 - 4, were resolved into four pairs of enantiomers, (+)-1 and (-)-1, (+)-2 and (-)-2, (+)-3 and (-)-3, and (+)-4 and (-)-4, by chiral HPLC. The structures and absolute configurations were elucidated on the basis of comprehensive spectroscopic analyses, quantum chemical calculations, quantitative measurements of molar rotations, application of van't Hoff's principle of optical superposition, and comparison with the literature data. The NMR and MS data of compound 3 are reported for the first time, as the structure was listed in SciFinder Scholar with no associated reference. These non-brominated pyrrole derivatives were found in this species for the first time. Compound 18 showed valuable cytotoxicities against HL-60, K562, and HCT-116 cell lines with IC50 values of 12.4, 16.0, and 19.8 μm, respectively. Compounds 16 - 19, 21, and 22 showed potent antifungal activities against Candida albicans with MIC values ranging from 0.59 to 4.69 μg/ml. Compounds 16 - 19 exhibited moderate antibacterial activities against Proteusbacillus vulgaris (MIC values ranging from 9.38 to 18.75 μg/ml). © 2017 Wiley-VHCA AG, Zurich, Switzerland.

  7. Identification and characterization of a novel diterpene gene cluster in Aspergillus nidulans.

    Directory of Open Access Journals (Sweden)

    Kirsi Bromann

    Full Text Available Fungal secondary metabolites are a rich source of medically useful compounds due to their pharmaceutical and toxic properties. Sequencing of fungal genomes has revealed numerous secondary metabolite gene clusters, yet products of many of these biosynthetic pathways are unknown since the expression of the clustered genes usually remains silent in normal laboratory conditions. Therefore, to discover new metabolites, it is important to find ways to induce the expression of genes in these otherwise silent biosynthetic clusters. We discovered a novel secondary metabolite in Aspergillus nidulans by predicting a biosynthetic gene cluster with genomic mining. A Zn(II(2Cys(6-type transcription factor, PbcR, was identified, and its role as a pathway-specific activator for the predicted gene cluster was demonstrated. Overexpression of pbcR upregulated the transcription of seven genes in the identified cluster and led to the production of a diterpene compound, which was characterized with GC/MS as ent-pimara-8(14,15-diene. A change in morphology was also observed in the strains overexpressing pbcR. The activation of a cryptic gene cluster by overexpression of its putative Zn(II(2Cys(6-type transcription factor led to discovery of a novel secondary metabolite in Aspergillus nidulans. Quantitative real-time PCR and DNA array analysis allowed us to predict the borders of the biosynthetic gene cluster. Furthermore, we identified a novel fungal pimaradiene cyclase gene as well as genes encoding 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA reductase and a geranylgeranyl pyrophosphate (GGPP synthase. None of these genes have been previously implicated in the biosynthesis of terpenes in Aspergillus nidulans. These results identify the first Aspergillus nidulans diterpene gene cluster and suggest a biosynthetic pathway for ent-pimara-8(14,15-diene.

  8. Characterization and Inhibition of a Class II Diterpene Cyclase from Mycobacterium tuberculosis

    Science.gov (United States)

    Mann, Francis M.; Prisic, Sladjana; Hu, Huayou; Xu, Meimei; Coates, Robert M.; Peters, Reuben J.

    2009-01-01

    Mycobacterium tuberculosis remains a widespread and devastating human pathogen, whose ability to infiltrate macrophage host cells from the human immune system is an active area of investigation. We have recently reported the discovery of a novel diterpene from M. tuberculosis, edaxadiene, whose ability to arrest phagosomal maturation in isolation presumably contributes to this critical process in M. tuberculosis infections. (Mann, F. M., Xu, M., Chen, X., Fulton, D. B., Russell, D. G., and Peters, R. J. (2009) J. Am. Chem. Soc., in press). Here, we present characterization of the class II diterpene cyclase that catalyzes the committed step in edaxadiene biosynthesis, i.e. the previously identified halimadienyl-diphosphate synthase (HPS; EC 5.5.1.16). Intriguingly, our kinetic analysis suggests a potential biochemical regulatory mechanism that triggers edaxadiene production upon phagosomal engulfment. Furthermore, we report characterization of potential HPS inhibitors: specifically, two related transition state analogs (15-aza-14,15-dihydrogeranylgeranyl diphosphate (7a) and 15-aza-14,15-dihydrogeranylgeranyl thiolodiphosphate (7b)) that exhibit very tight binding. Although arguably not suitable for clinical use, these nevertheless provide a basis for pharmaceutical design against this intriguing biosynthetic pathway. Finally, we provide evidence indicating that this pathway exists only in M. tuberculosis and is not functional in the closely related Mycobacterium bovis because of an inactivating frameshift in the HPS-encoding gene. Thus, we hypothesize that the inability to produce edaxadiene may be a contributing factor in the decreased infectivity and/or virulence of M. bovis relative to M. tuberculosis in humans. PMID:19574210

  9. Insights into Diterpene Cyclization from Structure of Bifunctional Abietadiene Synthase from Abies grandis*

    Science.gov (United States)

    Zhou, Ke; Gao, Yang; Hoy, Julie A.; Mann, Francis M.; Honzatko, Richard B.; Peters, Reuben J.

    2012-01-01

    Abietadiene synthase from Abies grandis (AgAS) is a model system for diterpene synthase activity, catalyzing class I (ionization-initiated) and class II (protonation-initiated) cyclization reactions. Reported here is the crystal structure of AgAS at 2.3 Å resolution and molecular dynamics simulations of that structure with and without active site ligands. AgAS has three domains (α, β, and γ). The class I active site is within the C-terminal α domain, and the class II active site is between the N-terminal γ and β domains. The domain organization resembles that of monofunctional diterpene synthases and is consistent with proposed evolutionary origins of terpene synthases. Molecular dynamics simulations were carried out to determine the effect of substrate binding on enzymatic structure. Although such studies of the class I active site do lead to an enclosed substrate-Mg2+ complex similar to that observed in crystal structures of related plant enzymes, it does not enforce a single substrate conformation consistent with the known product stereochemistry. Simulations of the class II active site were more informative, with observation of a well ordered external loop migration. This “loop-in” conformation not only limits solvent access but also greatly increases the number of conformational states accessible to the substrate while destabilizing the nonproductive substrate conformation present in the “loop-out” conformation. Moreover, these conformational changes at the class II active site drive the substrate toward the proposed transition state. Docked substrate complexes were further assessed with regard to the effects of site-directed mutations on class I and II activities. PMID:22219188

  10. Identification and Characterization of a Novel Diterpene Gene Cluster in Aspergillus nidulans

    Science.gov (United States)

    Bromann, Kirsi; Toivari, Mervi; Viljanen, Kaarina; Vuoristo, Anu; Ruohonen, Laura; Nakari-Setälä, Tiina

    2012-01-01

    Fungal secondary metabolites are a rich source of medically useful compounds due to their pharmaceutical and toxic properties. Sequencing of fungal genomes has revealed numerous secondary metabolite gene clusters, yet products of many of these biosynthetic pathways are unknown since the expression of the clustered genes usually remains silent in normal laboratory conditions. Therefore, to discover new metabolites, it is important to find ways to induce the expression of genes in these otherwise silent biosynthetic clusters. We discovered a novel secondary metabolite in Aspergillus nidulans by predicting a biosynthetic gene cluster with genomic mining. A Zn(II)2Cys6–type transcription factor, PbcR, was identified, and its role as a pathway-specific activator for the predicted gene cluster was demonstrated. Overexpression of pbcR upregulated the transcription of seven genes in the identified cluster and led to the production of a diterpene compound, which was characterized with GC/MS as ent-pimara-8(14),15-diene. A change in morphology was also observed in the strains overexpressing pbcR. The activation of a cryptic gene cluster by overexpression of its putative Zn(II)2Cys6–type transcription factor led to discovery of a novel secondary metabolite in Aspergillus nidulans. Quantitative real-time PCR and DNA array analysis allowed us to predict the borders of the biosynthetic gene cluster. Furthermore, we identified a novel fungal pimaradiene cyclase gene as well as genes encoding 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase and a geranylgeranyl pyrophosphate (GGPP) synthase. None of these genes have been previously implicated in the biosynthesis of terpenes in Aspergillus nidulans. These results identify the first Aspergillus nidulans diterpene gene cluster and suggest a biosynthetic pathway for ent-pimara-8(14),15-diene. PMID:22506079

  11. Central nervous system activities of two diterpenes isolated from Aloysia virgata.

    Science.gov (United States)

    Wasowski, Cristina; Marder, Mariel

    2011-03-15

    Using the guide of a competitive assay for the benzodiazepine binding site in the γ-aminobutyric acid type A receptor (GABA(A)), two active diterpenes were isolated from the aerial parts of Aloysia virgata (Ruíz & Pavón) A.L. Jussieu var. platyphylla (Briquet) Moldenke. These compounds, identified as (16R)-16,17,18-trihydroxyphyllocladan-3-one (1) and (16R)-16,17-dihydroxyphyllocladan-3-one (2) on the basis of spectral data, competitively inhibited the binding of [(3)H]-FNZ to the benzodiazepine binding site with K(i)±S.E.M. values of 56±19 μM and 111±13 μM, respectively. The behavioral actions of these diterpenes, intraperitoneally (i.p.) administered in mice, were examined in the plus-maze, holeboard, locomotor activity and light/dark tests. Compound 1 exhibited anxiolytic-like effects in mice evidenced by a significant increase of the parameters measured in the holeboard test (the number of head dips at 0.3 mg/kg and 3 mg/kg, the rears at 1 mg/kg and the time spent head-dipping at 3 mg/kg), in the plus-maze assay (the percentage of open arm entries at 1 mg/kg) and in the light/dark test (the time in light and the number of transitions at 1 mg/kg). Compound 2 augmented the number of rearings in the holeboard apparatus (at 0.3 mg/kg and 1 mg/kg) and the locomotor activity (at 1 mg/kg). These results reveal the presence of neuroactive compounds in Aloysia virgata.

  12. An ellagitannin, n-butyl gallate, two aryltetralin lignans, and an unprecedented diterpene ester from Pelargonium reniforme.

    Science.gov (United States)

    Latté, Klaus Peter; Kaloga, Maki; Schäfer, Andreas; Kolodziej, Herbert

    2008-02-01

    The structural diversity of the metabolic pool of Pelargonium reniforme was extended by the characterization of the 1C4-glucose based ellagitannin pelargoniin E, gallic acid n-butyl ester, (-)-4,4',9'-trihydroxy-3',5'-dimethoxy-2,7'-cyclolignan 9-O-beta-glucopyranoside and reniformin, a diterpene ester comprised of a diterpene acid with an uncommon -(CH2)(2)- bridging element linked to 2-(4-hydroxyphenyl)ethansulfonic acid. These metabolites were associated with the known (alpha,beta)-3,4-di-O-galloyl-glucopyranoside, 4,6-dihydroxy-2beta-glucopyranosyloxyacetophenone, 1-O-galloylglycerol, 6'-O-galloylsalidroside and (+)-isolariciresinol-9'-O-beta-glucopyranoside. All structures were established on the basis of spectroscopic methods.

  13. An Unusual Chimeric Diterpene Synthase from Emericella variecolor and Its Functional Conversion into a Sesterterpene Synthase by Domain Swapping.

    Science.gov (United States)

    Qin, Bin; Matsuda, Yudai; Mori, Takahiro; Okada, Masahiro; Quan, Zhiyang; Mitsuhashi, Takaaki; Wakimoto, Toshiyuki; Abe, Ikuro

    2016-01-26

    Di- and sesterterpene synthases produce C20 and C25 isoprenoid scaffolds from geranylgeranyl pyrophosphate (GGPP) and geranylfarnesyl pyrophosphate (GFPP), respectively. By genome mining of the fungus Emericella variecolor, we identified a multitasking chimeric terpene synthase, EvVS, which has terpene cyclase (TC) and prenyltransferase (PT) domains. Heterologous gene expression in Aspergillus oryzae led to the isolation of variediene (1), a novel tricyclic diterpene hydrocarbon. Intriguingly, in vitro reaction with the enzyme afforded the new macrocyclic sesterterpene 2 as a minor product from dimethylallyl pyrophosphate (DMAPP) and isopentenyl pyrophosphate (IPP). The TC domain thus produces the diterpene 1 and the sesterterpene 2 from GGPP and GFPP, respectively. Notably, a domain swap of the PT domain of EvVS with that of another chimeric sesterterpene synthase, EvSS, successfully resulted in the production of 2 in vivo as well. Cyclization mechanisms for the production of these two compounds are proposed.

  14. Dactyloditerpenol acetate, a new prenylbisabolane-type diterpene from Aplysia dactylomela with significant in vitro anti-neuroinflammatory activity

    Science.gov (United States)

    Jiménez-Romero, Carlos; Mayer, Alejandro M. S.; Rodríguez, Abimael D.

    2014-01-01

    A new regular diterpene possessing an unusual 1,6-anti-3-methylcyclohex-2-en-1-ol ring system, dactyloditerpenol acetate (1), has been extracted from the tropical sea hare Aplysia dactylomela and its stereostructure elucidated by spectroscopic methods. The absolute configuration of 1 was determined as 1S, 6S, 7R, 10S, and 11R by application of Kishi’s method for the assignment of absolute configuration of alcohols. The new diterpene potently inhibited in vitro thromboxane B2 (TXB2) (IC50 0.4 μM) and superoxide anion (O2−) (IC50 1 μM) generation from E. coli lipopolysaccharide (LPS)-activated rat neonatal microglia, with concomitant low short-term toxicity. PMID:24279991

  15. Trichocitrin, a new fusicoccane diterpene from the marine brown alga-endophytic fungus Trichoderma citrinoviride cf-27.

    Science.gov (United States)

    Liang, Xiao-Rui; Miao, Feng-Ping; Song, Yin-Ping; Guo, Zhan-Yong; Ji, Nai-Yun

    2016-07-01

    One new diterpene, trichocitrin (1), and four known secondary metabolites, nafuredin (2), 5-hydroxy-2,3-dimethyl-7-methoxychromone (3), 24-methylenecycloartanol (4) and citrostadienol (5), were isolated from the culture of marine brown alga-endophytic Trichoderma citrinoviride cf-27. Trichocitrin (1) represents the first Trichoderma-derived and furan-bearing fusicoccane diterpene, and its structure and relative configuration were identified by analysis of 1D/2D NMR and mass spectroscopic data. Compounds 1 and 2 exhibited 8.0- and 9.5-mm inhibition zones, respectively, against Escherichia coli at 20 μg/disc and 54.1 and 36.7% growth inhibition, respectively, of Prorocentrum donghaiense at 80 μg/mL.

  16. A clerodane diterpene inhibit adipogenesis by cell cycle arrest and ameliorate obesity in C57BL/6 mice.

    Science.gov (United States)

    Beg, Muheeb; Shankar, Kripa; Varshney, Salil; Rajan, Sujith; Singh, Suriya Pratap; Jagdale, Pankaj; Puri, Anju; Chaudhari, Bhushan P; Sashidhara, Koneni V; Gaikwad, Anil Nilkanth

    2015-01-01

    A clerodane diterpene, 16α-Hydroxycleroda-3, 13 (14) Z-dien-15, 16-olide (compound 1) isolated from Polyalthia longifolia had previously been reported as a new structural class of HMG-CoA reductase inhibitor apart from statins. Statins are known to be anti-adipogenic in nature. The distant structural similarity between compound 1 and lovastatin (polyketide class of compound) prompted us to investigate effects of diterpene compound 1 on adipogenesis and thereby obesity. High content microscopy proved diterpene compound 1 exhibits better anti-adipogenic activity and less toxicity in differentiating adipocytes. Moreover, it reduced expression levels of PPARγ, C/EBPα and GLUT4 during differentiation in a time and concentration dependent manner. Diterpene compound 1 during early differentiation reduced MDI induced-Akt/mTOR phosphorylation and expression of cell cycle proteins, and thereby halted mitotic clonal expansion, the decisive factor in early adipogenesis. Further, its anti-adipogenic activity was validated in murine mesenchymal cell-line C3H10T1/2 and human mesenchymal stem cell models of adipogenic differentiation. When compound 1 was administered along with HFD, for another 8 weeks in 2 month HFD fed overweight mice (with BMI > 30 and impaired glucose tolerance), it attenuated weight gain and epididymal fat accumulation. It improved body glucose tolerance, reduced HFD induced increase in total cholesterol and leptin/adiponectin ratio. All these effects were comparable with standard anti-obesity drug Orlistat with added edge of potently decreasing circulating triglyceride levels comparable with normal chow fed group. Histological analysis shows that compound 1 inhibit adipocyte hypertrophy and decreased steatosis in hepatocytes. Both in vivo and in vitro results demonstrate a potential value of compound 1 as a novel anti-adipogenic and anti-obesity agent.

  17. Determination of major phenolic acids, phenolic diterpenes and triterpenes in Rosemary (Rosmarinus Officinalis L.) by gas chromatography and mass spectrometry:

    OpenAIRE

    Vončina, Ernest; Doleček, Valter; Islamčević Razboršek, Maša; Brodnjak-Vončina, Darinka

    2007-01-01

    A gas chromatographic-mass spectrometric (GC-MS) method for the simultaneous identification and quantification of seven major phenolic and terpenic compounds in Rosmarinus officinalis L. was developed. The compounds were identified as trimethylsilyl (TMS) derivatives of phenolic acids (caffeic and rosmarinic acid), phenolic diterpene (carnosic acid), and pentacyclic triterpenes (ursolic, oleanolic, betulinic acid and betulin). These compounds have been identified by retention time and compari...

  18. Darwinolide, a New Diterpene Scaffold That Inhibits Methicillin-Resistant Staphylococcus aureus Biofilm from the Antarctic Sponge Dendrilla membranosa.

    Science.gov (United States)

    von Salm, Jacqueline L; Witowski, Christopher G; Fleeman, Renee M; McClintock, James B; Amsler, Charles D; Shaw, Lindsey N; Baker, Bill J

    2016-06-01

    A new rearranged spongian diterpene, darwinolide, has been isolated from the Antarctic Dendroceratid sponge Dendrilla membranosa. Characterized on the basis of spectroscopic and crystallographic analysis, the central seven-membered ring is hypothesized to originate from a ring-expansion of a spongian precursor. Darwinolide displays 4-fold selectivity against the biofilm phase of methicillin-resistant Staphylococcus aureus compared to the planktonic phase and may provide a scaffold for the development of therapeutics for this difficult to treat infection.

  19. Helikaurolides A-D with a Diterpene-Sesquiterpene Skeleton from Supercritical Fluid Extracts of Helianthus annuus L. var. Arianna.

    Science.gov (United States)

    Torres, Ascensión; Molinillo, José M G; Varela, Rosa M; Casas, Lourdes; Mantell, Casimiro; Martínez de la Ossa, Enrique J; Macías, Francisco A

    2015-10-02

    Four novel compounds (1-4) with an unprecedented skeleton that combines a sesquiterpene lactone and a kaurane diterpene acid were isolated from Helianthus annuus L. var. Arianna extract, which was obtained under supercritical conditions. The structures of 1-4 were elucidated by NMR and MS analyses. The biosynthetic routes involve sesquiterpene lactones and kauranic acid, both of which were previously isolated from this species.

  20. 卡萨烷型二萜的研究进展%Research Progress of Cassane Diterpene

    Institute of Scientific and Technical Information of China (English)

    唐洪波; 杨凤仙; 李元平; 王利勤

    2015-01-01

    卡萨烷型二萜主要分布在云实属植物中,本文对云实属以外的 Erythrophleum、Acacia、Myrospermum、Oryza、Chamaecrista、Swartzia、Osteospermum、Laxiflorus、Plathymenia、Bowdichia、Cordyla、Bobgunnia等属植物和Paraconiothyrium属真菌中的卡萨烷型二萜成分和生物活性研究现状进行综述.目前文献报道云实属植物以外资源中共有卡萨烷型二萜成分91个,它们的结构类型较多,有含氮(形成酰胺或胺)的卡萨烷二萜、卡萨烷型三环二萜、卡萨烷型呋喃二萜、具有内酯环的卡萨烷二萜、降卡萨烷二萜及二聚体等,活性也呈现多样性,如细胞毒性、抗菌、抗增殖等生物活性.%Cassane-type diterpenes are mainly distributed in Caesalpinia Plants. The cassane-type diterpenes from the plants of Erythrophleum, Acacia, Myrospermum, Oryza, Chamaecrista, Swartzia, Osteospermum, Laxiflorus, Plathymenia, Bowdichia, Cordyla and Bobgunnia genus, and the fungi of Paraconiothyrium genus were reviewed. These cassane diterpene showed cytotoxic, anti-bacterial, anti-proliferative and other biological activities.

  1. Modularity of Conifer Diterpene Resin Acid Biosynthesis: P450 Enzymes of Different CYP720B Clades Use Alternative Substrates and Converge on the Same Products1[OPEN

    Science.gov (United States)

    Yuen, Macaire M.S.; Bohlmann, Jörg

    2016-01-01

    Cytochrome P450 enzymes of the CYP720B subfamily play a central role in the biosynthesis of diterpene resin acids (DRAs), which are a major component of the conifer oleoresin defense system. CYP720Bs exist in families of up to a dozen different members in conifer genomes and fall into four different clades (I–IV). Only two CYP720B members, loblolly pine (Pinus taeda) PtCYP720B1 and Sitka spruce (Picea sitchensis) PsCYP720B4, have been characterized previously. Both are multisubstrate and multifunctional clade III enzymes, which catalyze consecutive three-step oxidations in the conversion of diterpene olefins to DRAs. These reactions resemble the sequential diterpene oxidations affording ent-kaurenoic acid from ent-kaurene in gibberellin biosynthesis. Here, we functionally characterized the CYP720B clade I enzymes CYP720B2 and CYP720B12 in three different conifer species, Sitka spruce, lodgepole pine (Pinus contorta), and jack pine (Pinus banksiana), and compared their activities with those of the clade III enzymes CYP720B1 and CYP720B4 of the same species. Unlike the clade III enzymes, clade I enzymes were ultimately found not to be active with diterpene olefins but converted the recently discovered, unstable diterpene synthase product 13-hydroxy-8(14)-abietene. Through alternative routes, CYP720B enzymes of both clades produce some of the same profiles of conifer oleoresin DRAs (abietic acid, neoabietic acid, levopimaric acid, and palustric acid), while clade III enzymes also function in the formation of pimaric acid, isopimaric acid, and sandaracopimaric acid. These results highlight the modularity of the specialized (i.e. secondary) diterpene metabolism, which produces conifer defense metabolites through variable combinations of different diterpene synthase and CYP720B enzymes. PMID:26936895

  2. Modularity of Conifer Diterpene Resin Acid Biosynthesis: P450 Enzymes of Different CYP720B Clades Use Alternative Substrates and Converge on the Same Products.

    Science.gov (United States)

    Geisler, Katrin; Jensen, Niels Berg; Yuen, Macaire M S; Madilao, Lina; Bohlmann, Jörg

    2016-05-01

    Cytochrome P450 enzymes of the CYP720B subfamily play a central role in the biosynthesis of diterpene resin acids (DRAs), which are a major component of the conifer oleoresin defense system. CYP720Bs exist in families of up to a dozen different members in conifer genomes and fall into four different clades (I-IV). Only two CYP720B members, loblolly pine (Pinus taeda) PtCYP720B1 and Sitka spruce (Picea sitchensis) PsCYP720B4, have been characterized previously. Both are multisubstrate and multifunctional clade III enzymes, which catalyze consecutive three-step oxidations in the conversion of diterpene olefins to DRAs. These reactions resemble the sequential diterpene oxidations affording ent-kaurenoic acid from ent-kaurene in gibberellin biosynthesis. Here, we functionally characterized the CYP720B clade I enzymes CYP720B2 and CYP720B12 in three different conifer species, Sitka spruce, lodgepole pine (Pinus contorta), and jack pine (Pinus banksiana), and compared their activities with those of the clade III enzymes CYP720B1 and CYP720B4 of the same species. Unlike the clade III enzymes, clade I enzymes were ultimately found not to be active with diterpene olefins but converted the recently discovered, unstable diterpene synthase product 13-hydroxy-8(14)-abietene. Through alternative routes, CYP720B enzymes of both clades produce some of the same profiles of conifer oleoresin DRAs (abietic acid, neoabietic acid, levopimaric acid, and palustric acid), while clade III enzymes also function in the formation of pimaric acid, isopimaric acid, and sandaracopimaric acid. These results highlight the modularity of the specialized (i.e. secondary) diterpene metabolism, which produces conifer defense metabolites through variable combinations of different diterpene synthase and CYP720B enzymes.

  3. Molecular Cloning and Functional Analysis of Gene Clusters for the Biosynthesis of Indole-Diterpenes in Penicillium crustosum and P. janthinellum

    Directory of Open Access Journals (Sweden)

    Matthew J. Nicholson

    2015-07-01

    Full Text Available The penitremane and janthitremane families of indole-diterpenes are abundant natural products synthesized by Penicillium crustosum and P. janthinellum. Using a combination of PCR, cosmid library screening, and Illumina sequencing we have identified gene clusters encoding enzymes for the synthesis of these compounds. Targeted deletion of penP in P. crustosum abolished the synthesis of penitrems A, B, D, E, and F, and led to accumulation of paspaline, a key intermediate for paxilline biosynthesis in P. paxilli. Similarly, deletion of janP and janD in P. janthinellum abolished the synthesis of prenyl-elaborated indole-diterpenes, and led to accumulation in the latter of 13-desoxypaxilline, a key intermediate for the synthesis of the structurally related aflatremanes synthesized by Aspergillus flavus. This study helps resolve the genetic basis for the complexity of indole-diterpene natural products found within the Penicillium and Aspergillus species. All indole-diterpene gene clusters identified to date have a core set of genes for the synthesis of paspaline and a suite of genes encoding multi-functional cytochrome P450 monooxygenases, FAD dependent monooxygenases, and prenyl transferases that catalyse various regio- and stereo- specific oxidations that give rise to the diversity of indole-diterpene products synthesized by this group of fungi.

  4. Effect of Diterpenes Isolated of the Marine Alga Canistrocarpus cervicornis against Some Toxic Effects of the Venom of the Bothrops jararaca Snake

    Directory of Open Access Journals (Sweden)

    Thaisa Francielle Souza Domingos

    2015-02-01

    Full Text Available Snake venoms are composed of a complex mixture of active proteins and peptides which induce a wide range of toxic effects. Envenomation by Bothrops jararaca venom results in hemorrhage, edema, pain, tissue necrosis and hemolysis. In this work, the effect of a mixture of two secodolastane diterpenes (linearol/isolinearol, previously isolated from the Brazilian marine brown alga, Canistrocarpus cervicornis, was evaluated against some of the toxic effects induced by B. jararaca venom. The mixture of diterpenes was dissolved in dimethylsulfoxide and incubated with venom for 30 min at room temperature, and then several in vivo (hemorrhage, edema and lethality and in vitro (hemolysis, plasma clotting and proteolysis assays were performed. The diterpenes inhibited hemolysis, proteolysis and hemorrhage, but failed to inhibit clotting and edema induced by B. jararaca venom. Moreover, diterpenes partially protected mice from lethality caused by B. jararaca venom. The search for natural inhibitors of B. jararaca venom in C. cervicornis algae is a relevant subject, since seaweeds are a rich and powerful source of active molecules which are as yet but poorly explored. Our results suggest that these diterpenes have the potential to be used against Bothropic envenomation accidents or to improve traditional treatments for snake bites.

  5. Molecular Cloning and Functional Analysis of Gene Clusters for the Biosynthesis of Indole-Diterpenes in Penicillium crustosum and P. janthinellum.

    Science.gov (United States)

    Nicholson, Matthew J; Eaton, Carla J; Stärkel, Cornelia; Tapper, Brian A; Cox, Murray P; Scott, Barry

    2015-07-23

    The penitremane and janthitremane families of indole-diterpenes are abundant natural products synthesized by Penicillium crustosum and P. janthinellum. Using a combination of PCR, cosmid library screening, and Illumina sequencing we have identified gene clusters encoding enzymes for the synthesis of these compounds. Targeted deletion of penP in P. crustosum abolished the synthesis of penitrems A, B, D, E, and F, and led to accumulation of paspaline, a key intermediate for paxilline biosynthesis in P. paxilli. Similarly, deletion of janP and janD in P. janthinellum abolished the synthesis of prenyl-elaborated indole-diterpenes, and led to accumulation in the latter of 13-desoxypaxilline, a key intermediate for the synthesis of the structurally related aflatremanes synthesized by Aspergillus flavus. This study helps resolve the genetic basis for the complexity of indole-diterpene natural products found within the Penicillium and Aspergillus species. All indole-diterpene gene clusters identified to date have a core set of genes for the synthesis of paspaline and a suite of genes encoding multi-functional cytochrome P450 monooxygenases, FAD dependent monooxygenases, and prenyl transferases that catalyse various regio- and stereo- specific oxidations that give rise to the diversity of indole-diterpene products synthesized by this group of fungi.

  6. Contrasting Patterns of Diterpene Acid Induction by Red Pine and White Spruce to Simulated Bark Beetle Attack, and Interspecific Differences in Sensitivity Among Fungal Associates.

    Science.gov (United States)

    Mason, Charles J; Klepzig, Kier D; Kopper, Brian J; Kersten, Philip J; Illman, Barbara L; Raffa, Kenneth F

    2015-06-01

    Conifers possess a suite of physiochemical defenses that protect their subcortical tissues from bark beetle - fungal complexes. These defenses include rapid induction of terpenoids and phenolics at the site of attack. Studies of the distribution, induction, and bioactivity of conifer terpenoids have focused heavily on monoterpenes. We assessed induction of diterpene acids in white spruce (Picea glauca) and red pine (Pinus resinosa) to fungal associates of two bark beetles, and the responses of four spruce beetle (Dendroctonus rufipennis)-associated fungi to three diterpene acids. Constitutive phloem contents differed between species, in that red pine had extremely low concentrations of diterpene acids, whereas white spruce had substantial constitutive levels. Induction differed quantitatively. Both red pine and white spruce exhibited marked increases, but red pine underwent greater increases and achieved higher concentrations than white spruce. Induction also differed qualitatively in that red pine showed lower diversity and fewer compositional changes during induction than white spruce. In red pine,fungal inoculation accompanying wounding elicited greater increases than wounding alone, but in white spruce total concentrations were higher following wounding alone. Spruce beetle fungal symbiont growth varied among species and compounds. Some diterpenes elicited both stimulatory and inhibitory effects on fungi, depending on concentration. All four fungi exhibited higher tolerances compared to those associated with pine bark beetles in previous studies. Variation in tolerances to, and potentially metabolism of, diterpene acids by symbionts may reflect differences in constitutive levels between spruce and pine, and partially explain differences in concentrations achieved during induction.

  7. Natural Diterpenes from Coffee, Cafestol, and Kahweol Induce Peripheral Antinoceception by Adrenergic System Interaction.

    Science.gov (United States)

    Guzzo, Luciana Souza; Castor, Marina Gomes Miranda E; Perez, Andrea de Castro; Duarte, Igor Dimitri Gama; Romero, Thiago Roberto Lima

    2016-01-01

    Cafestol and kahweol are diterpenes found only in the non-saponified lipid fraction of coffee. They are released during boiling and retained in the filtration process. Previous studies have shown peripheral antinociception induced by endogenous opioid peptides released by these diterpenes. Considering that the activation of the opioid system leads to a noradrenaline release, the aim of this study was to verify the participation of the noradrenergic system in the peripheral antinociception induced by cafestol and kahweol. Hyperalgesia was induced by an intraplantar injection of prostaglandin E2 (2 µg). Cafestol or kahweol (80 µg/paw) were administered locally into the right hindpaw alone, and after the agents α 2-adrenoceptor antagonist yohimbine (5, 10 and 20 µg/paw), α 2 A-adrenoceptor antagonist BRL 44 408 (40 µg/paw), α 2B-adrenoceptor antagonist imiloxan (40 µg/paw), α 2 C-adrenoceptor antagonist rauwolscine (10, 15 and 20 µg/paw), α 2D-adrenoceptor antagonist RX 821 002 (40 µg/paw), α 1-adrenoceptor antagonist prazosin (0.5, 1 and 2 µg/paw), or β-adrenoceptor antagonist propranolol (150, 300 and 600 ng/paw), respectively. Noradrenaline reuptake inhibitor reboxetine (30 µg/paw) was administered prior to cafestol or kahweol low dose (40 µg/paw) and guanetidine 3 days prior to the experiment (30 mg/kg, once a day), depleting the noradrenaline storage. Intraplantar injection of cafestol or kahweol (80 µg/paw) induced a peripheral antinociception against hyperalgesia induced by PGE2. This effect was reversed by intraplantar injections of yohimbine, rauwolscine, prazosin and propranolol. Reboxetine injection intensified the antinociceptive effect of cafestol or kahweol low-dose, and guanethidine reversed almost 70 % of the cafestol or kahweol-induced peripheral antinociception. This study gives evidence that the noradrenergic system participates in cafestol and kahweol-induced peripheral antinociception with the

  8. A new myrsinol-type diterpene polyester from Euphorbia dracunculoides Lam.

    Science.gov (United States)

    Wang, Li; Yang, Jing; Chi, Yu-Qian; Ouyang, Wen-Bin; Zang, Zhen; Huang, Sheng-Xiong; Cao, Pei; Zhao, Yong

    2015-01-01

    A new myrsinol-type diterpene polyester, 14-deoxo-3β-O-propinoyl-2α,5α,7β,15β-tetra-O-acetyl-14α-O-benzoyl-myrsinol (1), and its known analogue, 14-deoxo-3β-O-prorionyl-5α,15β-di-O-acetyl-7β-O-nicotinoyl-myrsinol-14β-acetate (2), together with a monoterpenoid, pubinernoid A (3), two indole alkaloids, neoechinulin A (4) and dihydroxyisoechinulin A (5), two benzene derivatives, siringin (6) and (3-methoxyphenyl) acetic acid (7), were isolated from the 70% acetone extract of the aerial parts of Euphorbia dracunculoides Lam. Their structures were elucidated on the basis of spectroscopic evidence and comparison with literature reports. The absolute configuration of 1 was deduced by comparing experimental and calculated ECD spectra. Among them, compounds 4 and 5 were first obtained from the plant source. In addition, the (13)C NMR data of compound 2 was reported for the first time.

  9. Modulation of inflammatory responses by diterpene acids from Helianthus annuus L.

    Science.gov (United States)

    Díaz-Viciedo, Rodrigo; Hortelano, Sonsoles; Girón, Natalia; Massó, Jose M; Rodriguez, Benjamín; Villar, Angel; de Las Heras, Beatriz

    2008-05-02

    Fractionation of a petroleum ether extract of Helianthus annuus L. led to the isolation of three diterpene acids: grandiflorolic, kaurenoic and trachylobanoic acids. These compounds were studied for potential anti-inflammatory activity on the generation of inflammatory mediators in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. At non-toxic concentrations, these compounds reduced, in a concentration-dependent manner nitric oxide (NO), prostaglandin E(2) (PGE(2)) and tumor necrosis factor (TNF-alpha) production, as well as expression of inducible nitric oxide synthase (NOS-2) and cyclooxygenase-2 (COX-2). All diterpenoids displayed significant in vivo anti-inflammatory activity and suppressed the 12-O-tetradecanoylphorbol-13-acetate (TPA)-mouse ear edema. In addition, inhibition of myeloperoxidase (MPO) activity, an index of cellular infiltration, was observed. In summary, our results suggest that the inhibition of the expression of NOS-2, COX-2 and the release of inflammatory cytokines, is responsible for the anti-inflammatory effects of the diterpenoids isolated from H. annuus L. which likely contributes to the pharmacological action of sunflower.

  10. A new acyclic diterpene acid and bioactive compounds from Knema glauca.

    Science.gov (United States)

    Rangkaew, Noppadon; Suttisri, Rutt; Moriyasu, Masataka; Kawanishi, Kazuko

    2009-05-01

    Investigation of the chemical constituents of the fruits of Knema glauca (Myristicaceae) yielded a new acyclic diterpene acid, named glaucaic acid 4, together with four acylphenols, including 1-(2,6-dihydroxyphenyl) tetradecan-1-one 1, malabaricone A 6, dodecanoylphloroglucinol 7 and 1-(2,4,6-trihydroxyphenyl)-9-phenylnonan-1-one 8, two lignans sesamin 2 and asarinin 3, and a flavan, myristinin D 5. In addition, myristinin A 9 and (+/-)-7,4'-dihydroxy-3'-methoxyflavan 10 were isolated from its leaves and stems, respectively. When tested against small-cell lung cancer (NCI-H187), epidermoid carcinoma (KB) and breast cancer (BC) cell lines, compounds 1, 6-8 and 10 displayed weak to moderate cytotoxicity. The acylphenols 6-8 displayed antituberculosis activity against the microbe Mycobacterium tuberculosis with MIC values of 25, 50 and 100 microg/mL, respectively, and antiviral activity against herpes simplex virus type 1, with 7 as the most active compound (IC(50) = 3.05 microg/mL). Malabaricone A 6 was also active against the malarial parasite Plasmodium falciparum with an IC(50) value of 2.78 microg/mL.

  11. Rebaudiosides R and S, Minor Diterpene Glycosides from the Leaves of Stevia rebaudiana.

    Science.gov (United States)

    Ibrahim, Mohamed A; Rodenburg, Douglas L; Alves, Kamilla; Perera, Wilmer H; Fronczek, Frank R; Bowling, John; McChesney, James D

    2016-05-27

    Two new diterpene glycosides have been isolated from a commercial extract of the leaves of Stevia rebaudiana. Compound 1 was shown to be 13-[(2-O-β-d-glucopyranosyl-3-O-β-d-glucopyranosyl-β-d-xylopyranosyl)oxy]ent-kaur-16-en-19-oic acid β-d-glucopyranosyl ester (rebaudioside R), while compound 2 was determined to be 13-[(2-O-α-d-glucopyranosyl-β-d-glucopyranosyl)oxy]ent-kaur-16-en-19-oic acid 2-O-α-l-rhamnopyranosyl-β-d-glucopyranosyl ester (rebaudioside S). Six additional known compounds were identified, dulcoside B, 13-[(2-O-β-d-xylopyranosyl-β-d-glucopyranosyl)oxy]ent-kaur-16-en-19-oic acid β-d-glucopyranosyl ester, eugenol diglucoside, rebaudioside G, 13-[(2-O-6-deoxy-β-d-glucopyranosyl-3-O-β-d-glucopyranosyl-β-d-glucopyranosyl)oxy]ent-kaur-16-en-19-oic acid β-d-glucopyranosyl ester, and rebaudioside D (3), respectively. The structures of 1 and 2 were determined based on comprehensive 1D and 2D NMR (COSY, HSQC, and HMBC) studies. A high-quality crystal of compound 3 allowed confirmation of its structure by X-ray diffraction.

  12. Furano diterpenes from Pterodon pubescens Benth with selective in vitro anticancer activity for prostate cell line

    Energy Technology Data Exchange (ETDEWEB)

    Spindola, Humberto M.; Carvalho, Joao E. de; Ruiz, Ana Lucia T.G.; Rodrigues, Rodney A. F.; Denny, Carina; Sousa, Ilza M. de Oliveira; Foglio, Mary Ann [Universidade Estadual de Campinas (UNICAMP), SP (Brazil). Centro Pluridisciplinar de Pesquisas Quimicas, Biologicas e Agricolas (CPQBA)]. E-mail: foglioma@cpqba.unicamp.br; Tamashiro, Jorge Y. [Universidade Estadual de Campinas (UNICAMP), SP (Brazil). Inst. de Biologia

    2009-07-01

    Activity guided fractionation of Pterodon pubescens Benth. methylene chloride-soluble fraction afforded novel 6{alpha}-acetoxi 7{beta}-hydroxy-vouacapan 1 and four known diterpene furans 2, 3, 4, 5. The compounds were evaluated for in vitro cytotoxic activities against human normal cells and tumour cell lines UACC-62 (melanoma), MCF-7 (breast), NCI-H460 (lung, non-small cells), OVCAR-03 (ovarian), PC-3 (prostate), HT-29 (colon), 786-0 (renal), K562 (leukemia) and NCI-ADR/RES (ovarian expressing phenotype multiple drugs resistance). Results were expressed by three concentration dependent parameters GI{sub 50} (concentration that produces 50% growth inhibition), TGI (concentration that produces total growth inhibition or cytostatic effect) and LC{sub 50} (concentration that produces .50% growth, a cytotoxicity parameter). Also, in vitro cytotoxicity was evaluated against 3T3 cell line (mouse embryonic fibroblasts). Antiproliferative properties of compounds 1, 4 and 5 are herein reported for the first time. These compounds showed selectivity in a concentration-dependent way against human PC-3. Compound 1 demonstrated selectivity 26 fold more potent than the positive control, doxorubicin, for PC-3 (prostrate) cell line based on GI{sub 50} values, causing cytostatic effect (TGI value) at a concentration fifteen times less than positive control. Moreover comparison of 50% lethal concentration (LC{sub 50} value) with positive control (doxorubicin) suggested that compound 1 was less toxic. (author)

  13. Manool, a Salvia officinalis diterpene, induces selective cytotoxicity in cancer cells.

    Science.gov (United States)

    de Oliveira, Pollyanna Francielli; Munari, Carla Carolina; Nicolella, Heloiza Diniz; Veneziani, Rodrigo Cassio Sola; Tavares, Denise Crispim

    2016-10-01

    Manool, a diterpene isolated from Salvia officinalis, was evaluated by the XTT colorimetric assay for cytotoxicity and selectivity against different cancer cell lines: B16F10 (murine melanoma), MCF-7 (human breast adenocarcinoma), HeLa (human cervical adenocarcinoma), HepG2 (human hepatocellular carcinoma), and MO59J, U343 and U251 (human glioblastoma). A normal cell line (V79, Chinese hamster lung fibroblasts) was used to compare the selectivity of the test substance. Manool exhibited higher cytotoxic activity against HeLa (IC50 = 6.7 ± 1.1 µg/mL) and U343 (IC50 = 6.7 ± 1.2 µg/mL) cells. In addition, in the used experimental protocols, the treatment with manool was significantly more cytotoxic for different tumor cell lines than for the normal cell line V79 (IC50 = 49.3 ± 3.3 µg/mL), and showed high selectivity. These results suggest that manool may be used to treat cancer without affecting normal cells.

  14. Three New Myrsinol Diterpenes from Euphorbia prolifera and Their Neuroprotective Activities

    Directory of Open Access Journals (Sweden)

    Yuanqiang Guo

    2012-08-01

    Full Text Available Three new myrsinol diterpenes were isolated from the roots of Euphorbia prolifera. Their structures were elucidated as 2α-O-isobutyryl-3β,5α,7β,10,15β-penta-O-acetyl-14α-O-benzoyl-10,18-dihydromyrsinol (1, 2α-O-isobutyryl-3β-O-propion-yl-5α,7β,10,15β-tetra-O-acetyl-10,18-dihydromyrsinol (2, and 2α,14α-di-O-benzoyl-3β,5α,7β,10,15β-penta-O-acetyl-10,18-dihydromyrsinol (3 on the basis of spectroscopic data analyses (IR, ESI-MS, HR-ESI-MS, and 1D and 2D NMR. Their neuroprotective activities were evaluated and compounds 1 and 2 showed neuroprotective effects against MPP+-induced neuronal cell death in SH-SY5Y cells.

  15. [Metabolic pathway and metabolites of total diterpene acid isolated from Pseudolarix kaempferi].

    Science.gov (United States)

    Liu, Peng; Guo, Hong-Zhu; Sun, Jiang-Hao; Xu, Man; Guo, Hui; Sun, Shi-Feng; Guo, De-An

    2014-08-01

    The preliminary metabolic profile of total diterpene acid (TDA) isolated from Pseudolarix kaempferi was investigated by using in vivo and in vitro tests. Pseudolaric acid C2 (PC2) was identified as the predominant metabolite in plasma, urine, bile and feces after both oral and intravenous administrations to rats using HPLC-UV and HPLC-ESI/MS(n), and demethoxydeacetoxypseudolaric acid B (DDPB), a metabolite proposed to be the glucoside of PC2 (PC2G), as well as pseudolaric acid C (PC), pseudolaric acid A (PA), pseudolaric acid A O-beta-D glucopyranoside (PAG), pseudolaric acid B O-beta-D glucopyranoside (PBG) and deacetylpseudolaric acid A (DPA) originated from TDA could also be detected. It was demonstrated by tests that the metabolism of TDA is independent of intestinal microflora, and neither of pepsin and trypsin is in charge of metabolism of TDA, TDA is also stable in both pH environments of gastric tract and intestinal tract. The metabolites of TDA in whole blood in vitro incubation were found to be PC2, DDPB and PC2G, which demonstrated that the metabolic reaction of TDA in vivo is mainly occurred in blood and contributed to be the hydrolysis of plasma esterase to ester bond, as well as the glucosylation reaction. These results clarified the metabolic pathway of TDA for the first time, which is of great significance to the in vivo active form and acting mechanism research of P. kaempferi.

  16. Structures of Some Novel α-Glucosyl Diterpene Glycosides from the Glycosylation of Steviol Glycosides

    Directory of Open Access Journals (Sweden)

    Indra Prakash

    2014-12-01

    Full Text Available Four new minor diterpene glycosides with a rare α-glucosyl linkage were isolated from a cyclodextrin glycosyltransferase glucosylated stevia extract containing more than 98% steviol glycosides. The new compounds were identified as 13-[(2-O-β-D-glucopyranosyl-3-O-(4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-β-D-glucopyranosyloxy] ent-kaur-16-en-19-oic acid-[(4-O-α-D-glucopyranosyl-β-D-glucopyranosyl ester] (1, 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyloxy] ent-kaur-16-en-19-oic acid-[(4-O-(4-O-(4-O-α-D-glucopyranosyl-α-D-glucopyranosyl-α-D-glucopyranosyl-β-D-glucopyranosyl ester] (2, 13-[(2-O-β-D-glucopyranosyl-3-O-(4-O-(4-O-(4-O-α-D-glucopyranosyl-α-D-glucopyranosyl-α-D-glucopyranosyl-β-D-glucopyranosyl-β-D-glucopyranosyloxy] ent-kaur-16-en-19-oic acid β-D-glucopyranosyl ester (3, and 13-[(2-O-β-D-glucopyranosyl-3-O-(4-O-(4-O-(4-O-α-D-glucopyranosyl-α-D-glucopyranosyl-α-D-glucopyranosyl-β-D-glucopyranosyl- β-D-glucopyranosyloxy] ent-kaur-16-en-19-oic acid-[(4-O-α-D-glucopyranosyl-β-D-glucopyranosyl ester] (4 on the basis of extensive NMR and mass spectral (MS data as well as hydrolysis studies.

  17. Overcoming the plasticity of plant specialized metabolism for selective diterpene production in yeast.

    Science.gov (United States)

    Ignea, Codruta; Athanasakoglou, Anastasia; Andreadelli, Aggeliki; Apostolaki, Maria; Iakovides, Minas; Stephanou, Euripides G; Makris, Antonios M; Kampranis, Sotirios C

    2017-08-18

    Plants synthesize numerous specialized metabolites (also termed natural products) to mediate dynamic interactions with their surroundings. The complexity of plant specialized metabolism is the result of an inherent biosynthetic plasticity rooted in the substrate and product promiscuity of the enzymes involved. The pathway of carnosic acid-related diterpenes in rosemary and sage involves promiscuous cytochrome P450s whose combined activity results in a multitude of structurally related compounds. Some of these minor products, such as pisiferic acid and salviol, have established bioactivity, but their limited availability prevents further evaluation. Reconstructing carnosic acid biosynthesis in yeast achieved significant titers of the main compound but could not specifically yield the minor products. Specific production of pisiferic acid and salviol was achieved by restricting the promiscuity of a key enzyme, CYP76AH24, through a single-residue substitution (F112L). Coupled with additional metabolic engineering interventions, overall improvements of 24 and 14-fold for pisiferic acid and salviol, respectively, were obtained. These results provide an example of how synthetic biology can help navigating the complex landscape of plant natural product biosynthesis to achieve heterologous production of useful minor metabolites. In the context of plant adaptation, these findings also suggest a molecular basis for the rapid evolution of terpene biosynthetic pathways.

  18. Formamido-Diterpenes from the South China Sea Sponge Acanthella cavernosa

    Directory of Open Access Journals (Sweden)

    Bao-Hua Zhang

    2012-07-01

    Full Text Available Seven new formamido-diterpenes, cavernenes A–D (14, kalihinenes E and F (56, and kalihipyran C (7, together with five known compounds (812, were isolated from the South China Sea sponge Acanthella cavernosa. Structures were established using IR, HRESIMS, 1D and 2D NMR, and single X-ray diffraction techniques. The isolated compounds were assessed for their cytotoxicity against a small panel of human cancer cell lines (HCT-116, A549, HeLa, QGY-7701, and MDA-MB-231 with IC50 values in the range of 6–18 μM. In addition, compound 9 showed weak antifungal activity against Trichophyton rubrum and Microsporum gypseum with MIC values of 8 and 32 μg/mL, respectively, compound 10 displayed weak antifungal activity against fungi Candida albicans, Cryptococcus neoformans, T. rubrum, and M. gypseum with MIC values of 8, 8, 4, and 8 μg/mL, respectively.

  19. Advanced Structural Determination of Diterpene Esters Using Molecular Modeling and NMR Spectroscopy.

    Science.gov (United States)

    Nothias-Scaglia, Louis-Félix; Gallard, Jean-François; Dumontet, Vincent; Roussi, Fanny; Costa, Jean; Iorga, Bogdan I; Paolini, Julien; Litaudon, Marc

    2015-10-23

    Three new jatrophane esters (1-3) were isolated from Euphorbia amygdaloides ssp. semiperfoliata, including an unprecedented macrocyclic jatrophane ester bearing a hemiketal substructure, named jatrohemiketal (3). The chemical structures of compounds 1-3 and their relative configurations were determined by spectroscopic analysis. The absolute configuration of compound 3 was determined unambiguously through an original strategy combining NMR spectroscopy and molecular modeling. Conformational search calculations were performed for the four possible diastereomers 3a-3d differing in their C-6 and C-9 stereocenters, and the lowest energy conformer was used as input structure for geometry optimization. The prediction of NMR parameters ((1)H and (13)C chemical shifts and (1)H-(1)H coupling constants) by density functional theory (DFT) calculations allowed identifying the most plausible diastereomer. Finally, the stereostructure of 3 was solved by comparison of the structural features obtained by molecular modeling for 3a-3d with NMR-derived data (the values of dihedral angles deduced from the vicinal proton-proton coupling constants ((3)JHH) and interproton distances determined by ROESY). The methodology described herein provides an efficient way to solve or confirm structural elucidation of new macrocyclic diterpene esters, in particular when no crystal structure is available.

  20. Antioxidant activity and mechanism of the abietane-type diterpene ferruginol.

    Science.gov (United States)

    Saijo, H; Kofujita, H; Takahashi, K; Ashitani, T

    2015-01-01

    The antioxidant activity of the abietane-type diterpene ferruginol was evaluated by comparison with that of carnosic acid, ( ± )-α-tocopherol and dibutylhydroxytoluene using 2,2-diphenyl-1-picrylhydrazyl, β-carotene bleaching and linoleic acid assays. Ferruginol had the lowest antioxidant activity of this group using the 2,2-diphenyl-1-picrylhydrazyl and β-carotene methods in polar solvent buffer. However, ferruginol exhibited stronger activity than carnosic acid and α-tocopherol for linoleic acid oxidation under non-solvent conditions. Five peaks corresponding to ferruginol derivatives were detected through GC-MS analysis of the reaction between ferruginol and methyl linoleate. The three reaction products were identified as dehydroferruginol, 7β-hydroxyferruginol and sugiol, and the other two peaks were assumed to be 7α-hydroxyferruginol and the quinone methide derivative of ferruginol. The time course of the reaction suggests that the quinone methide was produced early in the reaction and reacted further to produce dehydroferruginol, 7-hydroxyferruginol and sugiol. Thus, we inferred that quinone methide formation was a key step in the antioxidant reaction of ferruginol.

  1. Determination and potential importance of diterpene (kaur-16-ene) emitted from dominant coniferous trees in Japan.

    Science.gov (United States)

    Matsunaga, Sou N; Chatani, Satoru; Nakatsuka, Seiji; Kusumoto, Dai; Kubota, Katsuyoshi; Utsumi, Yasuhiro; Enoki, Tsutomu; Tani, Akira; Hiura, Tsutom

    2012-05-01

    Reactive volatile organic compounds (VOCs) are known to affect atmospheric chemistry. Biogenic VOCs (BVOCs) have a significant impact on regional air quality due to their large emission rates and high reactivities. Diterpenes (most particularly, kaur-16-ene) were detected in all of the 205 enclosure air samples collected over multiple seasons at two different sites from Cryptomeria japonica and Chamaecyparis obtusa trees, the dominant coniferous trees in Japan,. The emission rate of kaur-16-ene, was determined to be from 0.01 to 7.1 μg dwg(-1) h(-1) (average: 0.61 μg dwg(-1) h(-1)) employing branch enclosure measurements using adsorbent sampling followed by solid phase-liquid extraction techniques. The emission rate was comparable to that of monoterpenes, which is known major BVOC emissions, collected from the same branches. In addition, total emission of kaur-16-ene at 30°C was estimated to exceed that of total anthropogenic VOC emissions. Copyright © 2012 Elsevier Ltd. All rights reserved.

  2. Structures of some novel α-glucosyl diterpene glycosides from the glycosylation of steviol glycosides.

    Science.gov (United States)

    Prakash, Indra; Chaturvedula, Venkata Sai Prakash

    2014-12-04

    Four new minor diterpene glycosides with a rare α-glucosyl linkage were isolated from a cyclodextrin glycosyltransferase glucosylated stevia extract containing more than 98% steviol glycosides. The new compounds were identified as 13-[(2-O-β-D-glucopyranosyl-3-O-(4-O-α-D-glucopyranosyl)-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy] ent-kaur-16-en-19-oic acid-[(4-O-α-D-glucopyranosyl-β-D-glucopyranosyl) ester] (1), 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy] ent-kaur-16-en-19-oic acid-[(4-O-(4-O-(4-O-α-D-glucopyranosyl)-α-D-glucopyranosyl)-α-D-glucopyranosyl)-β-D-glucopyranosyl ester] (2), 13-[(2-O-β-D-glucopyranosyl-3-O-(4-O-(4-O-(4-O-α-D-glucopyranosyl)-α-D-glucopyranosyl)-α-D-glucopyranosyl)-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy] ent-kaur-16-en-19-oic acid β-D-glucopyranosyl ester (3), and 13-[(2-O-β-D-glucopyranosyl-3-O-(4-O-(4-O-(4-O-α-D-glucopyranosyl)-α-D-glucopyranosyl)-α-D-glucopyranosyl)-β-D-glucopyranosyl- β-D-glucopyranosyl)oxy] ent-kaur-16-en-19-oic acid-[(4-O-α-D-glucopyranosyl-β-D-glucopyranosyl) ester] (4) on the basis of extensive NMR and mass spectral (MS) data as well as hydrolysis studies.

  3. Intracellular Generation of a Diterpene-Peptide Conjugate that Inhibits 14-3-3-Mediated Interactions.

    Science.gov (United States)

    Parvatkar, Prakash; Kato, Nobuo; Uesugi, Motonari; Sato, Shin-Ichi; Ohkanda, Junko

    2015-12-23

    Synthetic agents that disrupt intracellular protein-protein interactions (PPIs) are highly desirable for elucidating signaling networks and developing new therapeutics. However, designing cell-penetrating large molecules equipped with the many functional groups necessary for binding to large interfaces remains challenging. Here, we describe a rational strategy for the intracellular oxime ligation-mediated generation of an amphipathic bivalent inhibitor composed of a peptide and diterpene natural product, fusicoccin, which binds 14-3-3 protein with submicromolar affinity. Our results demonstrate that co-treatment of cells with small module molecules, the aldehyde-containing fusicoccin 1 and the aminooxy-containing peptide 2, generates the corresponding conjugate 3 in cells, resulting in significant cytotoxicity. In contrast, chemically synthesized 3 is not cytotoxic, likely due to its inability to penetrate cells. Compound 3, but not 1 or 2, disrupts endogenous 14-3-3/cRaf interactions, suggesting that cell death is caused by inhibition of 14-3-3 activity. These results suggest that intracellular generation of large-sized molecules may serve as a new approach for modulating PPIs.

  4. The in vivo trypanocidal effect of the diterpene 5-epi-icetexone obtained from Salvia gilliesii.

    Science.gov (United States)

    Lozano, E; Strauss, M; Spina, R; Cifuente, D; Tonn, C; Rivarola, H W; Sosa, M A

    2016-02-01

    The search for new compounds with trypanocidal activity is crucial for the treatment of Chagas' disease. Previous in vitro studies have shown that the diterpene 5-epi-icetexone (ICTX) is active against Trypanosoma cruzi. The aim of this work was to evaluate the effect of ICTX on the parasites in infected mice, in an experimental model that mimics the acute phase of the disease. Swiss albino mice were infected with T. cruzi and treated daily with 10mg/kg/day ICTX (i.p.). Infected mice and mice injected with either saline or the vehicle DMSO were used as controls. Animals' survival and parasitemia were monitored once a week and histological studies were made at necropsy by the 5th week after infection. It was observed that the administration of ICTX increased the survival of mice infected, and induced a significant decrease in the parasitemia, as compared to controls. A similar protective effect was observed when animals were treated orally with benznidazole (BZN, used as a control of antiparasitic effect). By the 5th week post-infection, the presence of amastigote nests was observed within the fibers of the cardiac and skeletal muscle in controls, but not in animals treated with either ICTX or BZN. In addition, inflammatory infiltrates were observed in the tissues of controls, but not in animals treated with the drugs. We conclude that ICTX has an antiparasitic effect against T. cruzi, thus constituting an interesting option for the treatment of Chagas' disease, alone or combined with other drugs.

  5. Induction of Mitochondrial Dysfunction and Oxidative Stress in Leishmania donovani by Orally Active Clerodane Diterpene

    Science.gov (United States)

    Kathuria, Manoj; Bhattacharjee, Arindam; Sashidhara, Koneni V.; Singh, Suriya Pratap

    2014-01-01

    This study was performed to investigate the mechanistic aspects of cell death induced by a clerodane diterpene (K-09) in Leishmania donovani promastigotes that was previously demonstrated to be safe and orally active against visceral leishmaniasis (VL). K-09 caused depolarization of the mitochondrion and the generation of reactive oxygen species, triggering an apoptotic response in L. donovani promastigotes. Mitochondrial dysfunction subsequently resulted in the release of cytochrome c into the cytosol, impairing ATP production. Oxidative stress caused the depletion of reduced glutathione, while pretreatment with antioxidant N-acetyl cysteine (NAC) was able to abrogate oxidative stress. However, NAC failed to restore the mitochondrial membrane potential or intracellular calcium homeostasis after K-09 treatment, suggesting that the generation of oxidative stress is a downstream event relative to the other events. Caspase-3/-7-like protease activity and genomic DNA fragmentation were observed. Electron microscopy studies revealed gross morphological alterations typical of apoptosis, including severe mitochondrial damage, pyknosis of the nucleus, structural disruption of the mitochondrion-kinetoplast complex, flagellar pocket alterations, and the displacement of organelles. Moreover, an increased number of lipid droplets was detected after K-09 treatment, which is suggestive of altered lipid metabolism. Our results indicate that K-09 induces mitochondrial dysfunction and oxidative stress-mediated apoptotic cell death in L. donovani promastigotes, sharing many features with metazoan apoptosis. These mechanistic insights provide a basis for further investigation toward the development of K-09 as a potential drug candidate for VL. PMID:25070112

  6. Synthesis, Gastroprotective Effect and Cytotoxicity of New Amino Acid Diterpene Monoamides and Diamides

    Directory of Open Access Journals (Sweden)

    Daniel Droguett

    2010-10-01

    Full Text Available Following our studies on the gastroprotective effect and cytotoxicity of terpene derivatives, new amides were prepared from the diterpene 8(17-labden-15,19-dioic acid (junicedric acid and its 8(9-en isomer with C-protected amino acids (amino acid esters. The new compounds were evaluated for their gastroprotective effect in the ethanol/HCl-induced gastric lesions model in mice, as well as for cytotoxicity using the following human cell lines: normal lung fibroblasts (MRC-5, gastric adenocarcinoma cells (AGS and liver hepatocellular carcinoma (Hep G2. A dose-response experiment showed that at 25 mg/kg the C-15 leucyl and C-15,19-dileucylester amides of junicedric acid reduced gastric lesions by about 65.6 and 49.6%, respectively, with an effect comparable to lansoprazole at 20 mg/kg (79.3% lesion reduction. The comparison of the gastroprotective effect of 18 new amino acid ester amides was carried out at a single oral dose of 25 mg/kg. Several compounds presented a strong gastroprotective effect, reducing gastric lesions in the 70.9-87.8% range. The diprolyl derivative of junicedric acid, the most active product of this study (87.8% lesion reduction at 25 mg/kg presented a cytotoxicity value comparable with that of the reference compound lansoprazole. The structure-activity relationships are discussed.

  7. (+)-Dehydroabietic Acid, an Abietane-Type Diterpene, Inhibits Staphylococcus aureus Biofilms in Vitro

    Science.gov (United States)

    Fallarero, Adyary; Skogman, Malena; Kujala, Janni; Rajaratnam, Mohanathas; Moreira, Vânia M.; Yli-Kauhaluoma, Jari; Vuorela, Pia

    2013-01-01

    Potent drugs are desperately needed to counteract bacterial biofilm infections, especially those caused by gram-positive organisms, such as Staphylococcus aureus. Moreover, anti-biofilm compounds/agents that can be used as chemical tools are also needed for basic in vitro or in vivo studies aimed at exploring biofilms behavior and functionability. In this contribution, a collection of naturally-occurring abietane-type diterpenes and their derivatives was tested against S. aureus biofilms using a platform consisting of two phenotypic assays that have been previously published by our group. Three active compounds were identified: nordehydroabietylamine (1), (+)-dehydroabietic acid (2) and (+)-dehydroabietylamine (3) that prevented biofilm formation in the low micromolar range, and unlike typical antibiotics, only 2 to 4-fold higher concentrations were needed to significantly reduce viability and biomass of existing biofilms. Compound 2, (+)-dehydroabietic acid, was the most selective towards biofilm bacteria, achieving high killing efficacy (based on log Reduction values) and it was best tolerated by three different mammalian cell lines. Since (+)-dehydroabietic acid is an easily available compound, it holds great potential to be used as a molecular probe in biofilms-related studies as well as to serve as inspirational chemical model for the development of potent drug candidates. PMID:23739682

  8. (+-Dehydroabietic Acid, an Abietane-Type Diterpene, Inhibits Staphylococcus aureus Biofilms in Vitro

    Directory of Open Access Journals (Sweden)

    Pia Vuorela

    2013-06-01

    Full Text Available Potent drugs are desperately needed to counteract bacterial biofilm infections, especially those caused by gram-positive organisms, such as Staphylococcus aureus. Moreover, anti-biofilm compounds/agents that can be used as chemical tools are also needed for basic in vitro or in vivo studies aimed at exploring biofilms behavior and functionability. In this contribution, a collection of naturally-occurring abietane-type diterpenes and their derivatives was tested against S. aureus biofilms using a platform consisting of two phenotypic assays that have been previously published by our group. Three active compounds were identified: nordehydroabietylamine (1, (+-dehydroabietic acid (2 and (+-dehydroabietylamine (3 that prevented biofilm formation in the low micromolar range, and unlike typical antibiotics, only 2 to 4-fold higher concentrations were needed to significantly reduce viability and biomass of existing biofilms. Compound 2, (+-dehydroabietic acid, was the most selective towards biofilm bacteria, achieving high killing efficacy (based on log Reduction values and it was best tolerated by three different mammalian cell lines. Since (+-dehydroabietic acid is an easily available compound, it holds great potential to be used as a molecular probe in biofilms-related studies as well as to serve as inspirational chemical model for the development of potent drug candidates.

  9. [Study on limit detection of flavones in diterpene ginkgolides meglumine injection materials by LC-MS and HPLC-DAD].

    Science.gov (United States)

    Bi, Sen; Li, Yan-jing; Huang, Wen-zhe; Kang, Dan-yu; Ding, Gang; Xiao, Wei

    2015-08-01

    Limit test of flavones in diterpene ginkgolides meglumine injection materials by UV-Vis and HPLC-DAD method was studied in this essay. The HPLC-DAD method has lower LOD (about 1% of the UV-Vis), that is, the sensitivity is higher than UV-Vis method. Through the analysis of the kinds of flavonoids ingredients in the samples by LC-MS, the three compounds with highest contents are kaempferol, quercetin and isorhamnetin. Kaempferol, quercetin and isorhamnetin were chosen as reference compounds for HPLC analysis, and the HPLC separation analysis was carried on an Agilent Eclipse plus C18 column (4.6 mm x 250 mm, 5 μm) with methanol and water containing 0.4% phosphoric acid (50: 50) as mobile phase, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. This method has good specificity, precision and reproducibility. The LODs of quercetin, kaempferide and isorhamnetin were 27.6, 22.3, 29.5 μg x L(-1). The average recovery was 87.9% (RSD 3.3%), 91.7% (RSD 3.1%), 88.3 (RSD 1.3%) for quercetin, kaempferide and isorhamnetin, respectively. Based on the 10 batches of sample results and sensitivity of different HPLC, the content of total flavonoids ingredients of diterpene ginkgolides meglumine injection materials was limited no more than 2 x 10(-5). This method is simple, quick and has good maneuverability, and could be used to the limit test of flavonoids in the diterpene ginkgolides meglumine injection materials.

  10. Synthetic studies of neoclerodane diterpenes from Salvia divinorum: role of the furan in affinity for opioid receptors.

    Science.gov (United States)

    Simpson, Denise S; Lovell, Kimberly M; Lozama, Anthony; Han, Nina; Day, Victor W; Dersch, Christina M; Rothman, Richard B; Prisinzano, Thomas E

    2009-09-21

    Further synthetic modification of the furan ring of salvinorin A (1), the major active component of Salvia divinorum, has resulted in novel neoclerodane diterpenes with opioid receptor affinity and activity. A computational study has predicted 1 to be a reproductive toxicant in mammals and is suggestive that use of 1 may be associated with adverse effects. We report in this study that piperidine 21 and thiomorpholine 23 have been identified as selective partial agonists at kappa opioid receptors. This indicates that additional structural modifications of 1 may provide ligands with good selectivity for opioid receptors but with reduced potential for toxicity.

  11. A Synthetic Butenolide Diterpene is now a Natural Product Isolated from Metaporana sericosepala, a Plant from the Madagascar Dry Forest.

    Science.gov (United States)

    Presley, Christopher C; Rakotondraibe, L Harinantenaina; Brodie, Peggy J; Callmander, Martin W; Randrianaivo, Richard; Rasamison, Vincent E; Rakotobe, Etienne; Kingston, David G I

    2015-09-01

    Antiproliferative bioassay-guided fractionation of the ethanolic extract of the endemic Madagascan plant Metaporana sericosepala led to the first natural product isolation of a butenolide diterpene, which was synthesized during an anti-inflammatory study in 1988. The structure of the compound was elucidated as 3-homofarnesyl-4-hydroxybutenolide (1) by analysis of its spectroscopic data, including 1D- and 2D-NMR data and chemical evidence. The once synthetic compound can now also be considered as a natural product. Compound 1 had modest antiproliferative activity towards the A2780 ovarian cancer cell line,with an IC50 value of 8 µM.

  12. Diterpene alkaloids from roots of Aconitum ouvrardianum%德钦乌头中的二萜生物碱

    Institute of Scientific and Technical Information of China (English)

    丁祥; 侯怡铃

    2013-01-01

    目的 研究乌头属植物德钦乌头Aconitum ouvrardianum根的二萜生物碱类化学成分.方法 采用反复硅胶柱色谱进行分离纯化,根据谱学数据和对照TLC分析鉴定化合物的结构.结果 从德钦乌头根的醋酸乙酯提取物中分离得到12个化合物,分别鉴定为塔拉乌头胺(1)、彭乌碱乙(2)、卡马考宁(3)、黄草乌碱亭(4)、粗茎乌头碱甲(5)、贡乌生(6)、14-乙酰黄草乌碱亭(7)、展花乌头碱(8)、黄乌宁(9)、膝乌亭丁(10)、异塔拉乌头定(11)、翠雀它灵(12).结论 12个化合物均为C-3位或C-6位无含氧取代的C19-二萜生物碱,且均为首次从该植物中分离得到.乌头碱型C19-二萜生物碱为德钦乌头药材的主要药用化学成分.%Objective To study the diterpene alkaloids from the roots ofAconitum ouvrardianum.Methods The silica gel column was used for the isolation and purification of the compounds from the roots ofA.ouvrardianum.Their structures were identified on the bases of spectral data,such as NMR,TLC,and references.Results Twelve diterpene alkaloids:talatizamine (1),pengshenine B (2),cammaconine (3),sachaconitine (4),crassicauline (5),liljestrandisine (6),14-acetylsachaconitine (7),chasmaconitine (8),vilmorinine (9),geniculatine D (10),isotalatizidine (11),and detaline (12) were isolated and identified in the acetic ether extract from the roots of A.ouvrardianum.Conclusion All these compounds are C19-diterpene alkaloids without C-3 or C-6 oxygen substitution and are found in this plant for the first time.The C19-diterpene alkaloids are the main chemical constituents in A.ouvrardianum.

  13. The hallucinogenic diterpene salvinorin A inhibits leukotriene synthesis in experimental models of inflammation.

    Science.gov (United States)

    Rossi, Antonietta; Pace, Simona; Tedesco, Federica; Pagano, Ester; Guerra, Germano; Troisi, Fabiana; Werner, Markus; Roviezzo, Fiorentina; Zjawiony, Jordan K; Werz, Oliver; Izzo, Angelo A; Capasso, Raffaele

    2016-04-01

    Leukotrienes (LTs) are lipid mediators derived from arachidonic acid (AA) involved in a number of autoimmune/inflammatory disorders including asthma, allergic rhinitis and cardiovascular diseases. Salvinorin A (SA), a diterpene isolated from the hallucinogenic plant Salvia divinorum, is a well-established analgesic compound, but its anti-inflammatory properties are under-researched and its effects on LT production is unknown to date. Here, we studied the possible effect of SA on LT production and verified its actions on experimental models of inflammation in which LTs play a prominent role. Peritoneal macrophages (PM) stimulated by calcium ionophore A23187 were chosen as in vitro system to evaluate the effect of SA on LT production. Zymosan-induced peritonitis in mice and carrageenan-induced pleurisy in rats were selected as LT-related models to evaluate the effect of SA on inflammation as well as on LT biosynthesis. SA inhibited, in a concentration-dependent manner, A23187-induced LTB4 biosynthesis in isolated PM. In zymosan-induced peritonitis, SA inhibited cell infiltration, myeloperoxidase activity, vascular permeability and LTC4 production in the peritoneal cavity without decreasing the production of prostaglandin E2. In carrageenan-induced pleurisy in rats, a more sophisticated model of acute inflammation related to LTs, SA significantly inhibited LTB4 production in the inflammatory exudates, along with reducing the phlogistic process in the lung. In conclusion, SA inhibited LT production and it was effective in experimental models of inflammation in which LTs play a pivotal role. SA might be considered as a lead compound for the development of drugs useful in LTs-related diseases.

  14. Phytol, a diterpene alcohol from chlorophyll, as a drug against neglected tropical disease Schistosomiasis mansoni.

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    Josué de Moraes

    Full Text Available BACKGROUND: Schistosomiasis is a major endemic disease that affects hundreds of millions worldwide. Since the treatment and control of this parasitic disease rely on a single drug, praziquantel, it is imperative that new effective drugs are developed. Here, we report that phytol, a diterpene alcohol from chlorophyll widely used as a food additive and in medicinal fields, possesses promising antischistosomal properties in vitro and in a mouse model of schistosomiasis mansoni. METHODS AND FINDINGS: In vitro, phytol reduced the motor activity of worms, caused their death and confocal laser scanning microscopy analysis showed extensive tegumental alterations in a concentration-dependent manner (50 to 100 µg/mL. Additionally, phytol at sublethal doses (25 µg/mL reduced the number of Schistosoma mansoni eggs. In vivo, a single dose of phytol (40 mg/kg administered orally to mice infected with adult S. mansoni resulted in total and female worm burden reductions of 51.2% and 70.3%, respectively. Moreover, phytol reduced the number of eggs in faeces (76.6% and the frequency of immature eggs (oogram pattern was significantly reduced. The oogram also showed increases in the proportion of dead eggs. Confocal microcopy studies revealed tegumental damage in adult S. mansoni recovered from mice, especially in female worms. CONCLUSIONS: The significant reduction in parasite burden by this chlorophyll molecule validates phytol as a promising drug and offers the potential of a new direction for chemotherapy of human schistosomiasis. Phytol is a common food additive and nonmutagenic, with satisfactory safety. Thus, phytol has potential as a safe and cost-effective addition to antischistosomal therapy.

  15. Anti-angiogenic and anti-inflammatory properties of kahweol, a coffee diterpene.

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    Casimiro Cárdenas

    Full Text Available BACKGROUND: Epidemiological studies have shown that unfiltered coffee consumption is associated with a low incidence of cancer. This study aims to identify the effects of kahweol, an antioxidant diterpene contained in unfiltered coffee, on angiogenesis and key inflammatory molecules. METHODOLOGY/PRINCIPAL FINDINGS: The experimental procedures included in vivo angiogenesis assays (both the chicken and quail choriallantoic membrane assay and the angiogenesis assay with fluorescent zebrafish, the ex vivo mouse aortic ring assay and the in vitro analysis of the effects of treatment of human endothelial cells with kahweol in cell growth, cell viability, cell migration and zymographic assays, as well as the tube formation assay on Matrigel. Additionally, two inflammation markers were determined, namely, the expression levels of cyclooxygenase 2 and the levels of secreted monocyte chemoattractant protein-1. We show for the first time that kahweol is an anti-angiogenic compound with inhibitory effects in two in vivo and one ex vivo angiogenesis models, with effects on specific steps of the angiogenic process: endothelial cell proliferation, migration, invasion and tube formation on Matrigel. We also demonstrate the inhibitory effect of kahweol on the endothelial cell potential to remodel extracellular matrix by targeting two key molecules involved in the process, MMP-2 and uPA. Finally, the anti-inflammatory potential of this compound is demonstrated by its inhibition of both COX-2 expression and MCP-1 secretion in endothelial cells. CONCLUSION/SIGNIFICANCE: Taken together, our data indicate that, indeed, kahweol behaves as an anti-inflammatory and anti-angiogenic compound with potential use in antitumoral therapies. These data may contribute to the explanation of the reported antitumoral effects of kahweol, including the recent epidemiological meta-analysis showing that drinking coffee could decrease the risk of certain cancers.

  16. Anti-Angiogenic and Anti-Inflammatory Properties of Kahweol, a Coffee Diterpene

    Science.gov (United States)

    Cárdenas, Casimiro; Quesada, Ana R.; Medina, Miguel A.

    2011-01-01

    Background Epidemiological studies have shown that unfiltered coffee consumption is associated with a low incidence of cancer. This study aims to identify the effects of kahweol, an antioxidant diterpene contained in unfiltered coffee, on angiogenesis and key inflammatory molecules. Methodology/Principal Findings The experimental procedures included in vivo angiogenesis assays (both the chicken and quail choriallantoic membrane assay and the angiogenesis assay with fluorescent zebrafish), the ex vivo mouse aortic ring assay and the in vitro analysis of the effects of treatment of human endothelial cells with kahweol in cell growth, cell viability, cell migration and zymographic assays, as well as the tube formation assay on Matrigel. Additionally, two inflammation markers were determined, namely, the expression levels of cyclooxygenase 2 and the levels of secreted monocyte chemoattractant protein-1. We show for the first time that kahweol is an anti-angiogenic compound with inhibitory effects in two in vivo and one ex vivo angiogenesis models, with effects on specific steps of the angiogenic process: endothelial cell proliferation, migration, invasion and tube formation on Matrigel. We also demonstrate the inhibitory effect of kahweol on the endothelial cell potential to remodel extracellular matrix by targeting two key molecules involved in the process, MMP-2 and uPA. Finally, the anti-inflammatory potential of this compound is demonstrated by its inhibition of both COX-2 expression and MCP-1 secretion in endothelial cells. Conclusion/Significance Taken together, our data indicate that, indeed, kahweol behaves as an anti-inflammatory and anti-angiogenic compound with potential use in antitumoral therapies. These data may contribute to the explanation of the reported antitumoral effects of kahweol, including the recent epidemiological meta-analysis showing that drinking coffee could decrease the risk of certain cancers. PMID:21858104

  17. Phytol, a Diterpene Alcohol from Chlorophyll, as a Drug against Neglected Tropical Disease Schistosomiasis Mansoni

    Science.gov (United States)

    de Moraes, Josué; de Oliveira, Rosimeire N.; Costa, Jéssica P.; Junior, Antonio L. G.; de Sousa, Damião P.; Freitas, Rivelilson M.; Allegretti, Silmara M.; Pinto, Pedro L. S.

    2014-01-01

    Background Schistosomiasis is a major endemic disease that affects hundreds of millions worldwide. Since the treatment and control of this parasitic disease rely on a single drug, praziquantel, it is imperative that new effective drugs are developed. Here, we report that phytol, a diterpene alcohol from chlorophyll widely used as a food additive and in medicinal fields, possesses promising antischistosomal properties in vitro and in a mouse model of schistosomiasis mansoni. Methods and findings In vitro, phytol reduced the motor activity of worms, caused their death and confocal laser scanning microscopy analysis showed extensive tegumental alterations in a concentration-dependent manner (50 to 100 µg/mL). Additionally, phytol at sublethal doses (25 µg/mL) reduced the number of Schistosoma mansoni eggs. In vivo, a single dose of phytol (40 mg/kg) administered orally to mice infected with adult S. mansoni resulted in total and female worm burden reductions of 51.2% and 70.3%, respectively. Moreover, phytol reduced the number of eggs in faeces (76.6%) and the frequency of immature eggs (oogram pattern) was significantly reduced. The oogram also showed increases in the proportion of dead eggs. Confocal microcopy studies revealed tegumental damage in adult S. mansoni recovered from mice, especially in female worms. Conclusions The significant reduction in parasite burden by this chlorophyll molecule validates phytol as a promising drug and offers the potential of a new direction for chemotherapy of human schistosomiasis. Phytol is a common food additive and nonmutagenic, with satisfactory safety. Thus, phytol has potential as a safe and cost-effective addition to antischistosomal therapy. PMID:24392173

  18. Identification of diterpene alkaloids from Aconitum napellus subsp. firmum and GIRK channel activities of some Aconitum alkaloids.

    Science.gov (United States)

    Kiss, Tivadar; Orvos, Péter; Bánsághi, Száva; Forgo, Peter; Jedlinszki, Nikoletta; Tálosi, László; Hohmann, Judit; Csupor, Dezső

    2013-10-01

    Diterpene alkaloids neoline (1), napelline (2), isotalatizidine (3), karakoline (4), senbusine A (5), senbusine C (6), aconitine (7) and taurenine (8) were identified from Aconitum napellus L. subsp. firmum, four (2-4, 6) of which are reported for the first time from this plant. The structures were determined by means of LC-MS, 1D and 2D NMR spectroscopy, including (1)H-(1)H COSY, NOESY, HSQC and HMBC experiments. Electrophysiological effects of the isolated compounds, together with nine diterpene alkaloids previously obtained from Aconitum toxicum and Consolida orientalis were investigated on stable transfected HEK-hERG (Kv11.1) and HEK-GIRK1/4 (Kir3.1 and Kir3.4) cell lines using automated patch clamp equipment. Significant blocking activity on GIRK channel was exerted by aconitine (7) (45% at 10 μM), but no blocking activities of the other investigated compounds were detected. The tested compounds were inactive on hERG channel in the tested concentration. The comparison of the previously reported metabolites of A. napellus subsp. firmum and compounds identified in our experiment reveals substantial variability of the alkaloid profile of this taxon.

  19. In vitro cytotoxic activity of abietane diterpenes from Peltodon longipes as well as Salvia miltiorrhiza and Salvia sahendica.

    Science.gov (United States)

    Fronza, M; Murillo, R; Ślusarczyk, S; Adams, M; Hamburger, M; Heinzmann, B; Laufer, S; Merfort, I

    2011-08-15

    Phytochemical investigations of the n-hexane extract from the roots of Peltodon longipes (Lamiaceae) resulted in the isolation of 12 known abietane diterpenes (1-12). Structures were established on the basis of one and two dimensional nuclear magnetic resonance spectroscopic data ((1)H and (13)C, COSY, HSQC and HMBC), electron ionization mass spectrometric analysis (EIMS) as well as comparison with data from literature. These compounds, as well as eight known diterpenes (13-19) from Salvia miltiorrhiza, and two from Salvia sahendica (20 and 21) were evaluated for their cytotoxic effects in human pancreatic (MIAPaCa-2) and melanoma (MV-3) tumor cell lines using the MTT assay. Tanshinone IIa (13), 7α-acetoxyroyleanone (1), 1,2-dihydrotanshinone (16) and cryptotanshinone (14) had the highest cytotoxic effects in MIAPaCa-2, displaying IC(50) of 1.9, 4.7, 5.6, and 5.8 μM, respectively. Structure-activity relationships of abietane diterpenoid quinones are discussed.

  20. Direct and comprehensive analysis of ginsenosides and diterpene alkaloids in Shenfu injection by combinatory liquid chromatography-mass spectrometric techniques.

    Science.gov (United States)

    Yang, Hua; Liu, Lei; Gao, Wen; Liu, Ke; Qi, Lian-Wen; Li, Ping

    2014-04-01

    Shenfu injection (SFI) is a widely used Chinese herbal formulation for cardiac diseases prepared from red ginseng and processed aconite root. Clinical observations and pharmacological effects on SFI have been well investigated. Chemical analysis and quality control studies of this formulation, however, are relatively limited, especially regarding toxic aconite alkaloids. In this work, a high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-QTOF MS) method was applied to comprehensive analysis of constituents in SFI. Highly sensitive MS allows direct analysis of injections without additional sample pretreatment required. Using diagnostic ions and fragmentation rules, we identified 23 trace diterpene alkaloids, nineteen ginseng saponins, one panaxytriol, and one 5-hydroxymethylfurfural in SFI. A LC-MS method with selected ion monitoring was then used to quantify 24 major alkaloids and ginsenosides. The method was validated in terms of linearity, accuracy and precision. Especially, the limits of quantification were low to 0.4-18ng/mL for diterpene alkaloids. The total concentrations of saponins and alkaloids were about 676-742μg/mL and 3-7μg/mL in five batches of SFI samples, respectively. Finally, cosine ratio and euclidean distance were introduced to evaluate the batch-to-batch reproducibility of SFI samples, and the results demonstrated high quality consistency. Global identification and quantification of complex constituents based on LC-MS promises wide applications in quality control and batch monitoring for herbal products.

  1. Erythroivorensin: A novel anti-inflammatory diterpene from the root-bark of Erythrophleum ivorense (A Chev.).

    Science.gov (United States)

    Armah, Francis A; Annan, Kofi; Mensah, Abraham Y; Amponsah, Isaac K; Tocher, Derek A; Habtemariam, Solomon

    2015-09-01

    The stem- and root-bark of Erythrophleum ivorense (A Chev., family, Fabaceae) are routinely employed in the West African traditional medicine to treat inflammation and a variety of other disease conditions. Although the chemistry and pharmacology of cassaine-type diterpene alkaloids isolated from the stem-bark of the plant are fairly established, the root-bark has not yet been investigated. In the present study, the crude aqueous-alcohol extract of the root-bark was demonstrated to display a time- and dose (30-300 mg/kg p.o.)-dependent anti-inflammatory effect in chicks. Comprehensive chromatographic analysis coupled with spectroscopic and X-ray study further allowed the assignment of one of the major anti-inflammatory constituents as a novel cassaine-type diterpene, erythroivorensin. The other major constituents were known anti-inflammatory compounds: a triterpene, betulinic acid and a flavonoid, eriodictyol. The dose (10-100mg/kg p.o.)-dependent anti-inflammatory effects of the three compounds were either comparable or more significant than the positive control, diclofenac.

  2. A Pimarane Diterpene and Cytotoxic Angucyclines from a Marine-Derived Micromonospora sp. in Vietnam’s East Sea

    Science.gov (United States)

    Mullowney, Michael W.; Ó hAinmhire, Eoghainín; Tanouye, Urszula; Burdette, Joanna E.; Pham, Van Cuong; Murphy, Brian T.

    2015-01-01

    A screening of our actinomycete fraction library against the NCI-60 SKOV3 human tumor cell line led to the isolation of isopimara-2-one-3-ol-8,15-diene (1), lagumycin B (2), dehydrorabelomycin (3), phenanthroviridone (4), and WS-5995 A (5). These secondary metabolites were produced by a Micromonospora sp. isolated from sediment collected off the Cát Bà peninsula in the East Sea of Vietnam. Compound 1 is a novel Δ8,9-pimarane diterpene, representing one of approximately 20 actinomycete-produced diterpenes reported to date, while compound 2 is an angucycline antibiotic that has yet to receive formal characterization. The structures of 1 and 2 were elucidated by combined NMR and MS analysis and the absolute configuration of 1 was assigned by analysis of NOESY NMR and CD spectroscopic data. Compounds 2–5 exhibited varying degrees of cytotoxicity against a panel of cancerous and non-cancerous cell lines. Overall, this study highlights our collaborative efforts to discover novel biologically active molecules from the large, underexplored, and biodiversity-rich waters of Vietnam’s East Sea. PMID:26389922

  3. Indole Diterpene Alkaloids as Novel Inhibitors of the Wnt/β-catenin Pathway in Breast Cancer Cells

    Science.gov (United States)

    Sallam, Asmaa A.; Ayoub, Nehad M.; Foudah, Ahmed I.; Gissendanner, Chris R.; Meyer, Sharon A.; El Sayed, Khalid A.

    2013-01-01

    Penitrems are indole diterpene alkaloids best known for their BK channel inhibition and tremorgenic effects in mammals. In a previous study, penitrems A–F (1–5), their biosynthetic precursors, paspaline (6) and emindole SB (7), and two brominated penitrem analogs 8 and 9 demonstrated promising in vitro antiproliferative, antimigratory, and anti-invasive effects in the MTT (MCF-7 and MDA-MB-231), wound-healing, and Cultrex® BME cell invasion (MDA-MB-231) assays, respectively. The study herein reports the novel ability of penitrem A to suppress total β-catenin levels in MDA-MB-231 mammary cancer cells. Nine new penitrem analogs (10–18) were semisynthetically prepared, in an attempt to identify pharmacophores correlated with BK channel inhibition and tremorgenicity of penitrems and decrease their toxicity. The degree of BK channel inhibition was assessed using the nematode Caenorhabditis elegans, and in vivo tremorgenic EC50 was calculated using CD-1 male mice following an Up-and-Down-Procedure (UDP). Although new analogs were generally less active than parent compound 1, some showed no BK channel inhibition or tremorgenicity and retained the ability of penitrem A (1) to suppress total β-catenin levels in MDA-MB-231 cells. Paspaline (6) and emindole SB (7), both lacking BK channel inhibition and tremorgenicity, represent the simplest indole diterpene skeleton that retains the antiproliferative, antimigratory and total β-catenin suppressing effects shown by the more complex penitrem A (1). PMID:24211635

  4. A Pimarane Diterpene and Cytotoxic Angucyclines from a Marine-Derived Micromonospora sp. in Vietnam's East Sea.

    Science.gov (United States)

    Mullowney, Michael W; Ó hAinmhire, Eoghainín; Tanouye, Urszula; Burdette, Joanna E; Pham, Van Cuong; Murphy, Brian T

    2015-09-15

    A screening of our actinomycete fraction library against the NCI-60 SKOV3 human tumor cell line led to the isolation of isopimara-2-one-3-ol-8,15-diene (1), lagumycin B (2), dehydrorabelomycin (3), phenanthroviridone (4), and WS-5995 A (5). These secondary metabolites were produced by a Micromonospora sp. isolated from sediment collected off the Cát Bà peninsula in the East Sea of Vietnam. Compound 1 is a novel Δ(8,9)-pimarane diterpene, representing one of approximately 20 actinomycete-produced diterpenes reported to date, while compound 2 is an angucycline antibiotic that has yet to receive formal characterization. The structures of 1 and 2 were elucidated by combined NMR and MS analysis and the absolute configuration of 1 was assigned by analysis of NOESY NMR and CD spectroscopic data. Compounds 2-5 exhibited varying degrees of cytotoxicity against a panel of cancerous and non-cancerous cell lines. Overall, this study highlights our collaborative efforts to discover novel biologically active molecules from the large, underexplored, and biodiversity-rich waters of Vietnam's East Sea.

  5. A Pimarane Diterpene and Cytotoxic Angucyclines from a Marine-Derived Micromonospora sp. in Vietnam’s East Sea

    Directory of Open Access Journals (Sweden)

    Michael W. Mullowney

    2015-09-01

    Full Text Available A screening of our actinomycete fraction library against the NCI-60 SKOV3 human tumor cell line led to the isolation of isopimara-2-one-3-ol-8,15-diene (1, lagumycin B (2, dehydrorabelomycin (3, phenanthroviridone (4, and WS-5995 A (5. These secondary metabolites were produced by a Micromonospora sp. isolated from sediment collected off the Cát Bà peninsula in the East Sea of Vietnam. Compound 1 is a novel Δ8,9-pimarane diterpene, representing one of approximately 20 actinomycete-produced diterpenes reported to date, while compound 2 is an angucycline antibiotic that has yet to receive formal characterization. The structures of 1 and 2 were elucidated by combined NMR and MS analysis and the absolute configuration of 1 was assigned by analysis of NOESY NMR and CD spectroscopic data. Compounds 2–5 exhibited varying degrees of cytotoxicity against a panel of cancerous and non-cancerous cell lines. Overall, this study highlights our collaborative efforts to discover novel biologically active molecules from the large, underexplored, and biodiversity-rich waters of Vietnam’s East Sea.

  6. Design, synthesis and biological evaluation of tricyclic diterpene derivatives as novel neuroprotective agents against ischemic brain injury.

    Science.gov (United States)

    Wang, Ying-Ying; Gao, Yuan-Xue; Gao, Wei; Xu, Yuan; Xu, Ya-Zhou; Wang, Yun-Jie; Chang, Sai; Yu, Li-Gang; Zhang, Lu-Yong; Liao, Hong; Yang, Lian-Fang; Pang, Tao; Qiu, Wen-Wei

    2015-10-20

    Lead compound 7 has neuroprotective effects, and it was discovered by screening a small synthetic natural product-like (NPL) library. Based on the lead, a series of tricyclic diterpene derivatives was designed and synthesized, and their neuroprotective effects were further evaluated against glutamate-, oxygen and glucose deprivation (OGD)- and nutrient deprivation-induced neuronal injury using cell-based assays. To our delight, most of these synthetic compounds exhibited increased neuroprotective effects and blood-brain barrier (BBB) permeability without cellular toxicity. The most potent compound, compound 30, showed significantly improved neuroprotection against neuronal injury in primary neurons. Furthermore, compound 30 exhibited remarkable neuroprotection in transient middle cerebral artery occlusion (tMCAO) rats by reducing their infarct sizes and neurological deficit scores. A mechanistic exploration using in vitro and in vivo experiments showed that the neuroprotection of these compounds was at least partly mediated by improving the levels of glutathione (GSH), superoxide dismutase (SOD) and heme oxygenase-1 (HO-1) protein. Therefore, these tricyclic diterpene derivatives could be used as promising leads for the development of a new type of neuroprotective agents against ischemic brain injury.

  7. The coffee diterpene cafestol increases plasma triacylglycerol by increasing the production rate of large VLDL apolipoprotein B in healthy normolipidemic subjects

    NARCIS (Netherlands)

    Roos, de B.; Caslake, M.J.; Stalenhoef, A.F.H.; Bedford, D.; Demacker, P.N.; Katan, M.B.; Packard, C.J.

    2001-01-01

    Background: Cafestol is a diterpene in unfiltered coffee that raises plasma triacylglycerol in humans. Objective: We studied whether cafestol increases plasma triacylglycerol by increasing the production rate or by decreasing the fractional catabolic rate of VLDL1 [Svedberg flotation unit (Sf) 60-40

  8. Estudo multicasos sobre atividades inovativas

    Directory of Open Access Journals (Sweden)

    Sonia Regina Hierro Parolin

    2013-09-01

    Full Text Available Em estudos sobre competitividade argumenta-se que as empresas necessitam de estratégias baseadas em inovações e de capacidades internas em constantes e intensas transformações, não somente nos padrões tecnológicos, mas também no encadeamento do processo de gestão das atividades inovativas, como processos dinâmicos, não lineares e diversificados, em função de múltiplos fatores dos ambientes interno e externo. Para contribuir com essas discussões, neste artigo apresenta-se um estudo multicasos em quatro indústrias de médio e grande portes, de segmentos, densidades tecnológicas e históricos com inovações diferentes entre si. Como principais resultados, salientam-se o encadeamento das atividades inovativas como parte de uma estratégia organizacional para obter resultados com inovação e a afluência de todas as pessoas, e não somente as alocadas em pesquisa e desenvolvimento, para o cumprimento dessa estratégia.

  9. Evolution of Diterpene Metabolism: Sitka Spruce CYP720B4 Catalyzes Multiple Oxidations in Resin Acid Biosynthesis of Conifer Defense against Insects1[C][W][OA

    Science.gov (United States)

    Hamberger, Björn; Ohnishi, Toshiyuki; Hamberger, Britta; Séguin, Armand; Bohlmann, Jörg

    2011-01-01

    Diterpene resin acids (DRAs) are specialized (secondary) metabolites of the oleoresin defense of conifers produced by diterpene synthases and cytochrome P450s of the CYP720B family. The evolution of DRA metabolism shares common origins with the biosynthesis of ent-kaurenoic acid, which is highly conserved in general (primary) metabolism of gibberellin biosynthesis. Transcriptome mining in species of spruce (Picea) and pine (Pinus) revealed CYP720Bs of four distinct clades. We cloned a comprehensive set of 12 different Sitka spruce (Picea sitchensis) CYP720Bs as full-length cDNAs. Spatial expression profiles, methyl jasmonate induction, and transcript enrichment in terpenoid-producing resin ducts suggested a role of CYP720B4 in DRA biosynthesis. CYP720B4 was characterized as a multisubstrate, multifunctional enzyme by the formation of oxygenated diterpenoids in metabolically engineered yeast, yeast in vivo transformation of diterpene substrates, in vitro assays with CYP720B4 protein produced in Escherichia coli, and alteration of DRA profiles in RNA interference-suppressed spruce seedlings. CYP720B4 was active with 24 different diterpenoid substrates, catalyzing consecutive C-18 oxidations in the biosynthesis of an array of diterpene alcohols, aldehydes, and acids. CYP720B4 was most active in the formation of dehydroabietic acid, a compound associated with insect resistance of Sitka spruce. We identified patterns of convergent evolution of CYP720B4 in DRA metabolism and ent-kaurene oxidase CYP701 in gibberellin metabolism and revealed differences in the evolution of specialized and general diterpene metabolism in a gymnosperm. The genomic and functional characterization of the gymnosperm CYP720B family highlights that the evolution of specialized metabolism involves substantial diversification relative to conserved, general metabolism. PMID:21994349

  10. CYP99A3: Functional identification of a diterpene oxidase from the momilactone biosynthetic gene cluster in rice

    Science.gov (United States)

    Wang, Qiang; Hillwig, Matthew L.; Peters, Reuben J.

    2013-01-01

    SUMMARY Rice (Oryza sativa) produces momilactone diterpenoids as both phytoalexins and allelochemicals. Strikingly, the rice genome contains a biosynthetic gene cluster for momilactone production, located on rice chromosome 4, which contains two cytochromes P450 mono-oxygenases, CYP99A2 and CYP99A3, with undefined roles; although it has been previously shown that RNAi double knock-down of this pair of closely related CYP reduced momilactone accumulation. Here we attempted biochemical characterization of CYP99A2 and CYP99A3, which ultimately was achieved by complete gene recoding, enabling functional recombinant expression in bacteria. With these synthetic gene constructs it was possible to demonstrate that, while CYP99A2 does not exhibit significant activity with diterpene substrates, CYP99A3 catalyzes consecutive oxidations of the C19 methyl group of the momilactone precursor syn-pimara-7,15-diene to form, sequentially, syn-pimaradien-19-ol, syn-pimaradien-19-al and syn-pimaradien-19-oic acid. These are presumably intermediates in momilactone biosynthesis, as a C19 carboxylic acid moiety is required for formation of the core 19,6-γ-lactone ring structure. We further were able to detect syn-pimaradien-19-oic acid in rice plants, which indicates physiological relevance for the observed activity of CYP99A3. In addition, we found that CYP99A3 also oxidized syn-stemod-13(17)-ene at C19 to produce, sequentially, syn-stemoden-19-ol, syn-stemoden-19-al and syn-stemoden-19-oic acid, albeit with lower catalytic efficiency than with syn-pimaradiene. Although the CYP99A3 syn-stemodene derived products were not detected in planta, these results nevertheless provide a hint at the currently unknown metabolic fate of this diterpene in rice. Regardless of any wider role, our results strongly indicate that CYP99A3 acts as a multifunctional diterpene oxidase in momilactone biosynthesis. PMID:21175892

  11. Gastroprotective effect of epoxy clerodane diterpene isolated from Tinospora cordifolia Miers (Guduchi) on indomethacin-induced gastric ulcer in rats.

    Science.gov (United States)

    Antonisamy, Paulrayer; Dhanasekaran, Muniyappan; Ignacimuthu, Savarimuthu; Duraipandiyan, Veeramuthu; Balthazar, Joseph Devadass; Agastian, Paul; Kim, Jong-Hoon

    2014-06-15

    The present study evaluated the gastroprotective effect of epoxy clerodane diterpene (ECD), isolated from Tinospora cordifolia on indomethacin-induced gastric ulcer in rats. Administration of indomethacin exhibits extreme levels of ulcer index (UI) and myeloperoxidase (MPO) activity. Indomethacin down regulated PGE2, anti-inflammatory cytokines (IL-4, IL-10) and pro-angiogenic factors (VEGF and EGF). The ECD pretreatment considerably increased the levels of PGE2, anti-inflammatory cytokines and pro-angiogenic factors. The ulcer-healing activity of ECD was inhibited by pre-administration of the specific COX-1 inhibitor (SC560) and nonspecific NOS inhibitor (L-NAME), which indicates the involvement of PGE2 and NOS in ECD induced ulcer healing activity. These findings suggest that ECD exerts its antiulcer activity by reinforcement of defensive elements and diminishing the offensive elements.

  12. Chenopodolin: a phytotoxic unrearranged ent-pimaradiene diterpene produced by Phoma chenopodicola, a fungal pathogen for Chenopodium album biocontrol.

    Science.gov (United States)

    Cimmino, Alessio; Andolfi, Anna; Zonno, Maria C; Avolio, Fabiana; Santini, Antonello; Tuzi, Angela; Berestetskyi, Alexander; Vurro, Maurizio; Evidente, Antonio

    2013-07-26

    A new phytotoxic unrearranged ent-pimaradiene diterpene, named chenopodolin, was isolated from the liquid culture of Phoma chenopodicola, a fungal pathogen proposed for the biological control of Chenopodium album, a common worldwide weed of arable crops such as sugar beet and maize. The structure of chenopodolin was established by spectroscopic, X-ray, and chemical methods as (1S,2S,3S,4S,5S,9R,10S,12S,13S)-1,12-acetoxy-2,3-hydroxy-6-oxopimara-7(8),15-dien-18-oic acid 2,18-lactone. At a concentration of 2 mg/mL, the toxin caused necrotic lesions on Mercurialis annua, Cirsium arvense, and Setaria viride. Five derivatives were prepared by chemical modification of chenopodolin functionalities, and some structure-activity relationships are discussed.

  13. Andrographolide: A Novel Antimalarial Diterpene Lactone Compound from Andrographis paniculata and Its Interaction with Curcumin and Artesunate

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    Kirti Mishra

    2011-01-01

    Full Text Available Andrographolide (AND, the diterpene lactone compound, was purified by HPLC from the methanolic fraction of the plant Andrographis paniculata. The compound was found to have potent antiplasmodial activity when tested in isolation and in combination with curcumin and artesunate against the erythrocytic stages of Plasmodium falciparum in vitro and Plasmodium berghei ANKA in vivo. IC50s for artesunate (AS, andrographolide (AND, and curcumin (CUR were found to be 0.05, 9.1 and 17.4 μM, respectively. The compound (AND was found synergistic with curcumin (CUR and addictively interactive with artesunate (AS. In vivo, andrographolide-curcumin exhibited better antimalarial activity, not only by reducing parasitemia (29%, compared to the control (81%, but also by extending the life span by 2-3 folds. Being nontoxic to the in vivo system this agent can be used as template molecule for designing new derivatives with improved antimalarial properties.

  14. Andrographolide: A Novel Antimalarial Diterpene Lactone Compound from Andrographis paniculata and Its Interaction with Curcumin and Artesunate

    Science.gov (United States)

    Mishra, Kirti; Dash, Aditya P.; Dey, Nrisingha

    2011-01-01

    Andrographolide (AND), the diterpene lactone compound, was purified by HPLC from the methanolic fraction of the plant Andrographis paniculata. The compound was found to have potent antiplasmodial activity when tested in isolation and in combination with curcumin and artesunate against the erythrocytic stages of Plasmodium falciparum in vitro and Plasmodium berghei ANKA in vivo. IC50s for artesunate (AS), andrographolide (AND), and curcumin (CUR) were found to be 0.05, 9.1 and 17.4 μM, respectively. The compound (AND) was found synergistic with curcumin (CUR) and addictively interactive with artesunate (AS). In vivo, andrographolide-curcumin exhibited better antimalarial activity, not only by reducing parasitemia (29%), compared to the control (81%), but also by extending the life span by 2-3 folds. Being nontoxic to the in vivo system this agent can be used as template molecule for designing new derivatives with improved antimalarial properties. PMID:21760808

  15. Crystallization and preliminary X-ray diffraction analysis of the diterpene cyclooctatin synthase (CYC) from Streptomyces sp. LZ35

    Science.gov (United States)

    Zhang, Xiulei; Shang, Guijun; Gu, Lichuan; Shen, Yuemao

    2014-01-01

    Terpenoids are a large and highly diverse group of natural products, with the most chemically diverse pool of structures. Terpene synthase is the key enzyme in the process of terpenoid synthesis. In this paper, the first diterpene synthase (CYC) of bacterial origin was successfully crystallized. Native and SeMet-derivative crystals diffracted to 1.75 and 2.6 Å resolution, respectively. The native crystal belonged to space group P212121, with unit-cell parameters a = 59.10, b = 101.73, c = 108.93 Å, and contained two molecules per asymmetric unit. The SeMet-derivative crystal belonged to space group P21, with unit-cell parameters a = 58.64, b = 109.47, c = 58.73 Å, β = 119.35°, and had two molecules per asymmetric unit. PMID:24598929

  16. Simulations of circular dichroism spectra of a pair of diterpene enantiomers by time-dependent density functional theory

    Science.gov (United States)

    Liaw, Chih-Chuang; Chang, Jia-Lin; Chen, Shou-Fong; Huang, Jhih-Hong; Sie, Jyun-Fu; Cheng, Yung-Yi

    2011-11-01

    We present the first theoretical study on a pair of diterpene enantiomers of formula C 21H 34O 5, which were newly isolated from plants and identified as 3β,5β-dihydroxy-16 α/ β-methoxyhalima-13(14)-en-15,16-olide. The equilibrium geometries and harmonic vibrational frequencies of their low-lying conformers were obtained by using the AM1 and B3LYP/6-31+G(d) methods. At the optimized geometries, rotatory strengths of six excited states of each conformer were computed by the time-dependent density functional theory. The electronic circular dichroism spectra were simulated by taking Boltzmann averaging and considering the solvent effect, from which the absolute configurations of the enantiomers were determined. Their vibrational circular dichroism spectra were also predicted.

  17. A New Diterpene from Litsea cubeba Fruits: Structure Elucidation and Capability to Induce Apoptosis in HeLa Cells

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    Piyapat Trisonthi

    2014-05-01

    Full Text Available A new diterpene, identified as (+-6-(4-hydroxy-4-methyl-2-pentenoyl-4,6-dimethyl-5-(3-methyl-2-butenyl-1,3-cyclohexadienecarbaldehyde (1, cubelin, was isolated from a methanol extract of Litsea cubeba fruits by normal phase column chromatography and purified by preparative HPLC. The structure elucidation was conducted by spectroscopic methods (UV, IR, ESI-TOF-MS, 1-D and 2-D NMR. Cubelin exhibited activity against HeLa cell viability and proliferation. The cells also exhibited changes in nuclear morphology which are hallmarks of apoptotic cell death. The presence of cleaved caspase-3/-7, caspase-8 and caspase-9 in the cubelin treated population indicated the potential of the compound to induce apoptosis in HeLa cells via both intrinsic and extrinsic pathways.

  18. Indole diterpene alkaloids as novel inhibitors of the Wnt/β-catenin pathway in breast cancer cells.

    Science.gov (United States)

    Sallam, Asmaa A; Ayoub, Nehad M; Foudah, Ahmed I; Gissendanner, Chris R; Meyer, Sharon A; El Sayed, Khalid A

    2013-01-01

    Penitrems are indole diterpene alkaloids best known for their BK channel inhibition and tremorgenic effects in mammals. In a previous study, penitrems A-F (1-5), their biosynthetic precursors, paspaline (6) and emindole SB (7), and two brominated penitrem analogs 8 and 9 demonstrated promising in vitro antiproliferative, antimigratory, and anti-invasive effects in the MTT (MCF-7 and MDA-MB-231), wound-healing, and Cultrex BME cell invasion (MDA-MB-231) assays, respectively. The study herein reports the novel ability of penitrem A to suppress total β-catenin levels in MDA-MB-231 mammary cancer cells. Nine new penitrem analogs (10-18) were semisynthetically prepared, in an attempt to identify pharmacophores correlated with BK channel inhibition and tremorgenicity of penitrems and decrease their toxicity. The degree of BK channel inhibition was assessed using the nematode Caenorhabditis elegans, and in vivo tremorgenic EC₅₀ was calculated using CD-1 male mice following an Up-and-Down Procedure (UDP). Although new analogs were generally less active than parent compound 1, some showed no BK channel inhibition or tremorgenicity and retained the ability of penitrem A (1) to suppress total β-catenin levels in MDA-MB-231 cells. Paspaline (6) and emindole SB (7), both lacking BK channel inhibition and tremorgenicity, represent the simplest indole diterpene skeleton that retains the antiproliferative, antimigratory and total β-catenin suppressing effects shown by the more complex penitrem A (1). Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  19. Influence of ionic interactions on essential oil and phenolic diterpene composition of Dalmatian sage (Salvia officinalis L.).

    Science.gov (United States)

    Tounekti, Taieb; Munné-Bosch, Sergi; Vadel, A M; Chtara, Chaker; Khemira, Habib

    2010-01-01

    The potential of four essential cations (K(+), Ca(2+), Mg(2+) and Fe(2+)) to alleviate salt toxicity was studied in sage (Salvia officinalis L.) plants grown in pots. Two concentrations of the following chloride salts: KCl, CaCl₂, MgCl₂ and FeCl₃, were used together with 100mM NaCl to study the effects of these nutrients on plant growth, leaf essential oils (EOs) and phenolic diterpenes composition. The sage plants accumulated Na(+) in their leaves (includers); this has affected secondary metabolites' biosynthesis. Treatment with 100mM NaCl slightly decreased borneol and viridiflorol, while increased manool concentrations. Addition of KCl, CaCl₂ and MgCl₂ increased considerably in a dose-dependent manner the oxygen-containing monoterpenes (1.8-cineole, camphor, β-thujone and borneol) in 100mM NaCl-treated sage. Whereas, the contents of viridiflorol decreased further with the addition of KCl in 100mM NaCl-treated sage. Our results suggest that the changes in EOs composition were more related to K(+) and Ca(2+) availability than to Na(+) toxicity. Furthermore, treatment with NaCl decreased by 50% carnosic acid (CA), a potent antioxidant, content in the leaves. K(+) and Ca(2+) promoted the accumulation of CA and its methoxylated form (MCA) in the leaves. The concentration of CA was positively correlated with leaf K(+) (r=0.56, P=0.01) and Ca(2+) (r=0.44, P=0.05) contents. It appears that different salt applications in combination with NaCl treatments had a profound effect on EOs and phenolic diterpene composition in sage. Therefore, ionic interactions may be carefully considered in the cultivation of this species to get the desired concentrations of these secondary metabolites in leaf extracts.

  20. Improving the sensory and oxidative stability of cooked and chill-stored lamb using dietary rosemary diterpenes.

    Science.gov (United States)

    Serrano, Rafael; Ortuño, Jordi; Bañón, Sancho

    2014-09-01

    Two dietary rosemary extracts (DREs) containing diterpenes (carnosic acid and carnosol at 1:1 and 2:1 w:w) were tested in fattening lambs to stabilize the sensory quality of cooked and chill-stored patties. A total of 63 lambs were fed freely for 80 ± 5 d with a basal diet supplemented or not with DRE. Minced leg meat from each lamb was used to make patty batches. The patties were cooked at 72 ºC for 2 min, aerobically packed, kept at 2 ºC for up to 4 d and then reheated. Sensory traits (color, odor, flavor, and texture), CIELab color, and lipid oxidation (assessed as TBARS) were determined. In a first experiment, the lamb diet was supplemented with 600 mg of 1:1-DRE or 2:1-DRE kg(-1) feed. The 1:1-DRE diet delayed discoloration, flavor deterioration, and rancidity, while the 2:1-DRE diet was ineffective in this respect. In a second experiment, 4 supplementation levels of 1:1-DRE (0, 200, 400, and 600 mg kg(-1) feed) were compared. Flavor deterioration was delayed when the lamb diet was supplemented with at least 400 mg 1:1-DRE kg(-1) feed. The effects of the diet on the odor, flavor, and color were corroborated by differences in TBARS and CIELab. The results obtained suggest that rosemary diterpenes and/or their active secondary compounds deposited in muscle can act as endogenous antioxidants in cooked lamb. The carnosol intake seems crucial in the antioxidant actions achieved through DRE. The use of rosemary antioxidants in animal feeding would allow meat-based dishes to be preserved longer without adding preservatives.

  1. GC-MS-based metabolomic study on the antidepressant-like effects of diterpene ginkgolides in mouse hippocampus.

    Science.gov (United States)

    Liang, Zihong; Bai, Shunjie; Shen, Peng; Hu, Qingchuan; Wang, Xingfa; Dong, Meixue; Wang, Wei; Li, Juan; Cheng, Ke; Zhang, Shuxiao; Zou, Dezhi; Han, Yu; Wang, Haiyang; Xie, Peng

    2016-11-01

    Ginkgo biloba extract (GBE), including EGb-761, have been suggested to have antidepressant activity based on previous behavioral and biochemical analyses. However, because GBE contain many constituents, the mechanisms underlying this suggested antidepressant activity are unclear. Here, we investigated the antidepressant-like effects of diterpene ginkgolides (DG), an important class of constituents in GBE, and studied their effects in the mouse hippocampus using a GC-MS-based metabolomics approach. Mice were randomly divided into five groups and injected daily until testing with 0.9% NaCl solution, one of three doses of DG (4.06, 12.18, and 36.54mg/kg), or venlafaxine. Sucrose preference (SPT) and tail suspension (TST) tests were then performed to evaluate depressive-like behaviors in mice. DG (12.18 and 36.54mg/kg) and venlafaxine (VLX) administration significantly increased hedonic behavior in mice in the SPT. DG (12.18mg/kg) treatment also shortened immobility time in the TST, suggestive of antidepressant-like effects. Significant differences in the metabolic profile in the DG (12.18mg/kg) compared with the control or VLX group indicative of an antidepressant-like effect were observed using multivariate analysis. Eighteen differential hippocampal metabolites were identified that discriminated the DG (12.18mg/kg) and control groups. These biochemical changes involved neurotransmitter metabolism, oxidative stress, glutathione metabolism, lipid metabolism, energy metabolism, and kynurenic acid, providing clues to the therapeutic mechanisms of DG. Thus, this study showed that DG has antidepressant-like activities in mice and shed light on the biological mechanisms underlying the effects of diterpene ginkgolides on behavior, providing an important drug candidate for the treatment of depression.

  2. Determination of Total Diterpenes of Caesalpinia minax%苦石莲中总二萜的含量测定

    Institute of Scientific and Technical Information of China (English)

    韩瑞亭; 万国盛; 孙博航; 霍晓灵; 高慧媛

    2013-01-01

    目的:建立苦石莲中总二萜的含量测定方法,优化总二萜的提取工艺,并进行含量测定.方法:采用可见分光光度法,以5%香草醛/冰乙酸-高氯酸为显色剂,化合物caesalpinins K为对照品,在530 nm处测定其吸光度值,利用回归方程计算苦石莲中总二萜含量.结果:Caesalpinins K对照品溶液质量浓度在2.0~6.1 mg·L-1,线性关系良好,r =0.999 5(n=5),平均回收率为101.8% (RSD 2.3%).结论:该方法简便、准确、重复性好,可以为苦石莲的质量控制提供依据.%Objective: To establish quantitative methods of total diterpene in Caesalpinia minax and optimization of extraction process. Method; The content of total diterpene was determined at 530 nm by vis-spectrophotometry using caesalpinins K as standard and using the color reaction with 5% vanillina-glacial acetic acid and perchloric acid. Result; Linear relationship of caesalpinins K reference solution was good in the range of 2. 0-6.1 mg·L-1 (r = 0. 999 5) , the average recovery was 101. 8% ( RSD 2. 3% ). Conclusion: The present methods are simple, accurate with good reproducibility, and could be used to control the quality of Caesalpinia minax.

  3. Comparison of essential oils from Cistus species growing in Sardinia.

    Science.gov (United States)

    Mastino, Patrizia Monica; Marchetti, Mauro; Costa, Jean; Usai, Marianna

    2017-02-01

    Cistus genus is present in Sardinia with large populations of C. monspeliensis, C. salvifolius, C. creticus subsp. eriocephalus and few stations of C. albidus, C. creticus subsp. creticus and C. creticus subsp. corsicus. No chemical studies are currently being carried on Cistus species of Sardinia. The essential oils have shown six different profiles. C. creticus subsp. eriocephalus showed a high amount of manoyl oxide and its isomer (70%). C. salvifolius has pointed out the group of labdans, (20%); another consistent percentage is made of perfumed molecules as ionone and its derivate. Several linear hydrocarbons were produced by C. monspeliensis, and the heneicosane was the most represented element. In C. albidus no labdane-type diterpenes were identified. Analysis of C. creticus subsp creticus revealed several oxygenated sesquiterpenes and labdane-type diterpenes, especially manoyl oxide. C. creticus subsp. corsicus was qualitatively very similar to C. creticus subsp. creticus, notably concerning the labdane-type compounds.

  4. Isolation and HPLC quantitation of kaurane-type diterpenes and cinnamic acid derivatives of long-term stored leaves of Mikania laevigata and Mikania glomerata

    Directory of Open Access Journals (Sweden)

    SUZAN K.V. BERTOLUCCI

    2013-06-01

    Full Text Available The leaves of Mikania laevigata and Mikania glomerata are used in Brazil to treat respiratory affections, being kaurane-type diterpenes and coumarin considered as the bioactive compounds. The present study reports an investigation on the HPLC-DAD profiles and contents of coumarin (1, trans-o-coumaric (2, kaurenoic (3, benzoylgrandifloric (4 and cinnamoylgrandifloric (5 acids in dried leaves of Mikania species stored in dark room under controlled conditions. Excepting 2, the constituents were isolated and purified to be employed as reference compounds. The samples were analyzed at three monthly intervals up to 18 months for M. laevigata and 12 months for M. glomerata. trans-o-Coumaric was not detected in both, whereas 1 occurred only in M. laevigata. The concentrations of the assayed constituents did not vary significantly within the evaluated period (p < 0.05, for both species. In contrast, changes in the chromatographic profiles and spectral purity of peaks from 3, 4 and 5 were detected in samples of both Mikania stored for three months, while the coumarin profile in M. laevigata modified after six months of storage. The evaluation of chromatographic profiles based on spectral purity analyses of selected peaks was shown to be a more robust tool to access chemical stability of Mikania samples than the quantitation of chemical markers' contents.As folhas de Mikania laevigata e Mikania glomerata são usadas no Brasil para tratar afecções respiratórias, sendo diterpernos caur ânicos e cumarina considerados os constituintes bioativos. O presente estudo relata a investigação dos perfis por CLAE-DAD e os teores de cumarina (1, ácidos trans-o-coumárico (2, caurenóico (3, benzoilgrandiflórico (4 e cinamoilgrandiflórico (5 em folhas secas de espécies de Mikania armazenadas em sala escura, sob condições controladas. Com exceção de 2, os constituintes foram isolados e purificados para serem empregados como subst âncias de referência. As

  5. Jatrophane diterpenes as modulators of multidrug resistance. Advances of structure-activity relationships and discovery of the potent lead pepluanin A.

    Science.gov (United States)

    Corea, Gabriella; Fattorusso, Ernesto; Lanzotti, Virginia; Motti, Riccardo; Simon, Pierre-Noël; Dumontet, Charles; Di Pietro, Attilio

    2004-02-12

    From the whole plant of Euphorbia peplus L., five new diterpenes based on a jatrophane skeleton (pepluanins A-E, 1-5) were isolated, together with two known analogues (6 and 7), which served to divulge in detail the structure-activity relationships within this class of P-glycoprotein inhibitors. The results revealed the importance of substitutions on the medium-sized ring (carbons 8, 9, 14, and 15). In particular, the activity is collapsed by the presence of a free hydroxyl at C-8, while it increases with a carbonyl at C-14, an acetoxyl at C-9, and a free hydroxyl at C-15. The most potent compound of the series, pepluanin A, showed a very high activity for a jatrophane diterpene, outperforming cyclosporin A by a factor of at least 2 in the inhibition of Pgp-mediated daunomycin transport.

  6. Synthetic studies of neoclerodane diterpenes from Salvia divinorum: semisynthesis of salvinicins A and B and other chemical transformations of salvinorin A.

    Science.gov (United States)

    Harding, Wayne W; Schmidt, Matthew; Tidgewell, Kevin; Kannan, Pavitra; Holden, Kenneth G; Gilmour, Brian; Navarro, Hernan; Rothman, Richard B; Prisinzano, Thomas E

    2006-01-01

    Salvinorin A (1) is a hallucinogenic neoclerodane diterpene isolated from the widely available psychoactive plant Salvia divinorum and is the first example of a non-nitrogenous opioid receptor ligand. At present, there is little information available as to why this compound is selective for kappa opioid receptors. One approach to better understanding the mode of binding of 1 at kappa receptors is to systematically alter the structure of 1 and examine the effects on opioid receptor affinity and activity. Currently, there is a paucity of methods described for the preparation of analogues derived from 1. Here, we report the investigation of several chemical transformations of 1 isolated from S. divinorum. In particular, this work provides a semisynthesis of salvinicins A (2) and B (3) and has identified 10a as the first neoclerodane diterpene with delta opioid antagonist activity.

  7. 3-Acetylmaoecrystal S, a New Diterpene from Isodon eriocalyx%毛萼香茶菜中的一个新二萜

    Institute of Scientific and Technical Information of China (English)

    徐亮; 季红; 陈巧鸿; 陈东林; 简锡贤; 王锋鹏

    2006-01-01

    New diterpene 3-acetylmacoecrystal S (1) was isolated from Isodon eriocalyx (Dunn) Kudo, together with seven known diterpenes maoecrystals A, B, C and D, trichrokaurin, edonicin, and eriocalyxin B. The structure of 1 was elucidated on the basis of spectral evidence.%从毛萼香茶菜中分得8个二萜,3-乙酰毛萼结晶S、毛萼结晶甲、毛萼结晶乙、毛萼结晶丙、毛萼结晶丁、trichrokaurin、edonicin和eriocalyxin B.其中3-乙酰毛萼结晶S为新的对映-贝壳杉烷型二萜.由波谱法确定其结构.

  8. Antifeedant activity of anticopalic acid isolated from Vitex hemsleyi.

    Science.gov (United States)

    Villegas Gómez, Clarisa; Martínez-Vázquez, Mariano; Esquivel, Baldomero

    2009-01-01

    The known labdane-type diterpenoids anticopalic acid (1) and 3 beta-hydroxyanticopalic acid (2) were isolated from extracts of the aerial parts of Vitex hemsleyi Briq. (Labiatae). The acid 1 showed an antifeedant, dose-dependent activity against Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae). To our knowledge this is the first report on the antifeedant activity of a labdane-type diterpene against S. frugiperda.

  9. Disturbios da olfacao: estudo retrospectivo

    Directory of Open Access Journals (Sweden)

    Luciano Lobato Gregorio

    2014-01-01

    Full Text Available Introdução: O olfato, fenômeno subjetivo de grande importância, é pouco compreendido e estudado no ser humano. Médicos com maior conhecimento sobre os distúrbios desse sentido tendem a considerar a doença mais importante e manejar melhor o diagnóstico e o tratamento. Objetivo: Descrever a amostra dos pacientes com queixa principal de distúrbios do olfato e mostrar a experiência do serviço no manejo e tratamento. Delineamento: Estudo retrospectivo de coorte histórica com corte transversal. Materiais e métodos: Descrição da amostra e avaliação de resposta ao tratamento de pacientes com queixa principal de hiposmia ou anosmia atendidos no ambulatório de Rinologia no período de janeiro de 2005 a outubro de 2011. Resultados: Dos 38 pacientes com distúrbio da olfação, 68,4% dos pacientes apresentaram queixa de hiposmia e 31,5% de anosmia, com duração média de 30,8 meses. Os diagnósticos etiológicos principais foram idiopática (31,5%, rinopatia alérgica (28,9% e RSC com pólipos (10,5%. As respostas ao tratamento com corticosteroide tópico e ácido alfa-lipoico foram variáveis, assim como na literatura. Conclusão: Maior importância deve ser dada aos distúrbios do olfato na prática do otorrinolaringologista, uma vez que o diagnóstico diferencial é amplo e pode trazer grande morbidade ao paciente, com impacto na sua qualidade de vida.

  10. NaCl stress-induced changes in the essential oil quality and abietane diterpene yield and composition in common sage

    Directory of Open Access Journals (Sweden)

    Taieb Tounekti

    2015-09-01

    Full Text Available Aim: The purpose of this study was to evaluate how increasing NaCl salinity in the medium can affects the essential oils (EOs composition and phenolic diterpene content and yield in leaves of Salvia officinalis L. The protective role of such compounds against NaCl stress was also argued with regard to some physiological characteristics of the plant (water and ionic relations as well as the leaf gas exchanges. Materials and Methods: Potted plants were exposed to increasing NaCl concentrations (0, 50, 75 and 100 mM for 4 weeks during July 2012. Replicates from each treatment were harvested after 0, 2, 3 and 4 weeks of adding salt to perform physiological measurements and biochemical analysis. Results: Sage EOs were rich in manool, viridiflorol, camphor, and borneol. Irrigation with a solution containing 100 mM NaCl for 4 weeks increased considerably 1.8-cineole, camphor and beta-thujone concentrations, whereas lower concentrations (50 and 75 mM had no effects. On the contrary, borneol and viridiflorol concentrations decreased significantly under the former treatment, while manool and total fatty acid concentrations were not affected. Leaf extracts contained also several diterpenes such as carnosic acid (CA, carnosol (CAR and 12- and #1054;-methoxy carnosic acid (MCA. The concentrations and total contents of CA and MCA increased after 3 weeks of irrigation with 75 or 100 mM NaCl. The 50 mM NaCl had no effect on these diterpenes. Our results suggest a protective role for CA against salinity stress. Conclusion: This study may provide ways to manipulate the concentration and yield of some phenolic diterpenes and EOs in sage. In fact soil salinity may favour a directional production of particular components of interest. [J Intercult Ethnopharmacol 2015; 4(3.000: 208-216

  11. Teotihuacanin, a Diterpene with an Unusual Spiro-10/6 System from Salvia amarissima with Potent Modulatory Activity of Multidrug Resistance in Cancer Cells.

    Science.gov (United States)

    Bautista, Elihú; Fragoso-Serrano, Mabel; Toscano, Rubén A; García-Peña, María del Rosario; Ortega, Alfredo

    2015-07-01

    Teotihuacanin (1), an unusual rearranged clerodane diterpene with a new carbon skeleton containing a spiro-10/6 bicyclic system, was isolated from the leaves and flowers of Salvia amarissima. Its structure was determined through spectroscopic analyses. Its absolute configuration was established by single-crystal X-ray diffraction. Compound 1 showed potent modulatory activity of multidrug resistance in vinblastine-resistant MCF-7 cancer cell line (reversal fold, RFMCF-7/Vin+ > 10703) at 25 μg/mL.

  12. Nasute termite soldier frontal gland secretions. 1. Structure of Trinervi-2. beta. ,3. cap alpha. ,9. cap alpha. -triol 9-O acetate, a novel diterpene from Trinervitermes soldiers

    Energy Technology Data Exchange (ETDEWEB)

    Prestwich, G.D. (International Center of Insect Physiology and Ecology, Nairobi, Kenya); Tanis, S.P.; Springer, J.P.; Clardy, J.

    1976-09-15

    One of the major constituents, ''TG-2,'' of the soldier secretions of Trinervitermes gratiosus Sjostedt were studied by x-ray diffraction. The structure was determined to be a novel diterpene skeleton with a bridgehead double bond in an 11-membered ring. Bond angles and lengths were discussed, and a computer generated perspective drawing of the compound was presented. (DDA)

  13. Insights on profiling of phorbol, deoxyphorbol, ingenol and jatrophane diterpene esters by high performance liquid chromatography coupled to multiple stage mass spectrometry.

    Science.gov (United States)

    Nothias-Scaglia, Louis-Félix; Schmitz-Afonso, Isabelle; Renucci, Franck; Roussi, Fanny; Touboul, David; Costa, Jean; Litaudon, Marc; Paolini, Julien

    2015-11-27

    This paper reports our effort to develop a comprehensive HPLC-MS(n)-based dereplication strategy for phorbol ester (PE), deoxyphorbol ester (dPE) and ingenol ester (IE) profiling in plant extracts. This strategy is composed of two sequential analysis exploiting specific hybrid triple quadrupole/linear ion trap instrument modes. A first run was performed using a multiple reaction monitoring (MRM) mode targeting fragmentation of PE and dPE/IE coupled with the acquisition of MS(2) spectrum for the ions at m/z 311 and m/z 313, respectively. A second run was then completed based on precursor ion scan mode (PIS) and automatic MS(2) acquisition for each quasimolecular ion. The developed approach was used to investigate ten Euphorbia extracts showing bioactivity against chikungunya virus replication. Experiments allowed partial annotation of three dPE/IE but no PE was detected. Results suggested that other types of diterpene esters displayed PE- and dPE/IE-like fragmentations. The study of jatrophane ester (JE) standards by CID fragmentation using low and high resolution mass spectrometry confirmed this hypothesis, highlighting challenges and difficulties of diterpene esters profiling within plant extracts. Nonetheless, the present LC-MS(n) method can be easily adapted to profile other types of diterpene esters.

  14. Methylerythritol and mevalonate pathway contributions to biosynthesis of mono-, sesqui-, and diterpenes in glandular trichomes and leaves of Stevia rebaudiana Bertoni.

    Science.gov (United States)

    Wölwer-Rieck, Ursula; May, Bianca; Lankes, Christa; Wüst, Matthias

    2014-03-19

    The biosynthesis of the diterpenoid steviol glycosides rebaudioside A and stevioside in nonrooted cuttings of Stevia rebaudiana was investigated by feeding experiments using the labeled key precursors [5,5-(2)H2]-mevalonic acid lactone (d2-MVL) and [5,5-(2)H2]-1-deoxy-d-xylulose (d2-DOX). Labeled glycosides were extracted from the leaves and stems and were directly analyzed by LC-(-ESI)-MS/MS and by GC-MS after hydrolysis and derivatization of the resulting isosteviol to the corresponding TMS-ester. Additionally, the incorporation of the proffered d2-MVL and d2-DOX into volatile monoterpenes, sesquiterpenes, and diterpenes in glandular trichomes on leaves and stems was investigated by headspace-solid phase microextraction-GC-MS (HS-SPME-GC-MS). Incorporation of the labeled precursors indicated that diterpenes in leaves and monoterpenes and diterpenes in glandular trichomes are predominately biosynthesized via the methylerythritol phosphate (MEP) pathway, whereas both the MEP and mevalonate (MVA) pathways contribute to the biosynthesis of sesquiterpenes at equal rates in glandular trichomes. These findings give evidence for a transport of MEP pathway derived farnesyl diphosphate precursors from plastids to the cytosol. Contrarily, the transport of MVA pathway derived geranyl diphosphate and geranylgeranyl diphosphate precursors from the cytosol to the plastid is limited.

  15. Identification of a new cyathane diterpene that induces mitochondrial and autophagy-dependent apoptosis and shows a potent in vivo anti-colorectal cancer activity.

    Science.gov (United States)

    He, Luwei; Han, Junjie; Li, Baowei; Huang, Li; Ma, Ke; Chen, Quan; Liu, Xinzhong; Bao, Li; Liu, Hongwei

    2016-03-23

    Diterpenes has been reported to possess multiple bioactivities consisting of anti-microbial and anti-inflammatory properties. This study reveals a new cyathane-type diterpene (cyathin Q) from the culture of the fungus Cyathus africanus by bioactivity-guided separation. The structure of cyathin Q was determined based on spectroscopic measurements (NMR and MS). The bioactivity evaluation shows that cyathin Q has a strong anticancer activity against HCT116 cells and Bax-deficient HCT116 in vitro and in vivo. This compound induced hallmarks of apoptotic events in HCT116 cells, including caspase activation, cytochrome c release, poly (ADP-ribose) polymerase (PARP) cleavage, and depolarization of the mitochondrial inner transmembrane potential. This process is accompanied with the increased mitochondrial ROS, down-regulation of Bcl-2 protein, and up-regulation of Bim protein. We also observed the cleavage of autophagy-related protein ATG5 in cyathin Q-induced apoptosis. Taken together, our study identified a new fungal diterpene that exhibited anticancer activity via induction of mitochondria and autophagy-dependent apoptosis in HCT116 cells.

  16. Monoester-Diterpene Aconitum Alkaloid Metabolism in Human Liver Microsomes: Predominant Role of CYP3A4 and CYP3A5

    Science.gov (United States)

    Ye, Ling; Yang, Xiao-Shan; Lu, Lin-lin; Chen, Wei-Ying; Zeng, Shan; Yan, Tong-Meng; Dong, Ling-Na; Peng, Xiao-Juan; Shi, Jian; Liu, Zhong-Qiu

    2013-01-01

    Aconitum, widely used to treat rheumatoid arthritis for thousands of years, is a toxic herb that can frequently cause fatal cardiac poisoning. Aconitum toxicity could be decreased by properly hydrolyzing diester-diterpene alkaloids into monoester-diterpene alkaloids. Monoester-diterpene alkaloids, including benzoylaconine (BAC), benzoylmesaconine (BMA), and benzoylhypaconine (BHA), are the primary active and toxic constituents of processed Aconitum. Cytochrome P450 (CYP) enzymes protect the human body by functioning as the defense line that limits the invasion of toxicants. Our purposes were to identify the CYP metabolites of BAC, BMA, and BHA in human liver microsomes and to distinguish which isozymes are responsible for their metabolism through the use of chemical inhibitors, monoclonal antibodies, and cDNA-expressed CYP enzyme. High-resolution mass spectrometry was used to characterize the metabolites. A total of 7, 8, and 9 metabolites were detected for BAC, BMA, and BHA, respectively. The main metabolic pathways were demethylation, dehydrogenation, demethylation-dehydrogenation, hydroxylation and didemethylation, which produced less toxic metabolites by decomposing the group responsible for the toxicity of the parent compound. Taken together, the results of the chemical inhibitors, monoclonal antibodies, and cDNA-expressed CYP enzymes experiments demonstrated that CYP3A4 and CYP3A5 have essential functions in the metabolism of BAC, BMA, and BHA. PMID:23864901

  17. Monoester-Diterpene Aconitum Alkaloid Metabolism in Human Liver Microsomes: Predominant Role of CYP3A4 and CYP3A5

    Directory of Open Access Journals (Sweden)

    Ling Ye

    2013-01-01

    Full Text Available Aconitum, widely used to treat rheumatoid arthritis for thousands of years, is a toxic herb that can frequently cause fatal cardiac poisoning. Aconitum toxicity could be decreased by properly hydrolyzing diester-diterpene alkaloids into monoester-diterpene alkaloids. Monoester-diterpene alkaloids, including benzoylaconine (BAC, benzoylmesaconine (BMA, and benzoylhypaconine (BHA, are the primary active and toxic constituents of processed Aconitum. Cytochrome P450 (CYP enzymes protect the human body by functioning as the defense line that limits the invasion of toxicants. Our purposes were to identify the CYP metabolites of BAC, BMA, and BHA in human liver microsomes and to distinguish which isozymes are responsible for their metabolism through the use of chemical inhibitors, monoclonal antibodies, and cDNA-expressed CYP enzyme. High-resolution mass spectrometry was used to characterize the metabolites. A total of 7, 8, and 9 metabolites were detected for BAC, BMA, and BHA, respectively. The main metabolic pathways were demethylation, dehydrogenation, demethylation-dehydrogenation, hydroxylation and didemethylation, which produced less toxic metabolites by decomposing the group responsible for the toxicity of the parent compound. Taken together, the results of the chemical inhibitors, monoclonal antibodies, and cDNA-expressed CYP enzymes experiments demonstrated that CYP3A4 and CYP3A5 have essential functions in the metabolism of BAC, BMA, and BHA.

  18. Lolitrem B and Indole Diterpene Alkaloids Produced by Endophytic Fungi of the Genus Epichloë and Their Toxic Effects in Livestock.

    Science.gov (United States)

    Philippe, Guerre

    2016-02-15

    Different group of alkaloids are produced during the symbiotic development of fungal endophytes of the genus Epichloë in grass. The structure and toxicity of the compounds vary considerably in mammalian herbivores and in crop pests. Alkaloids of the indole-diterpene group, of which lolitrem B is the most toxic, were first characterized in endophyte-infected perennial ryegrass, and are responsible for "ryegrass staggers." Ergot alkaloids, of which ergovaline is the most abundant ergopeptide alkaloid produced, are also found in ryegrass, but generally at a lower rate than lolitrem B. Other alkaloids such as lolines and peramine are toxic for crop pests but have weak toxicological properties in mammals. The purpose of this review is to present indole-diterpene alkaloids produced in endophyte infected ryegrass from the first characterization of ryegrass staggers to the determination of the toxicokinetics of lolitrem B and of their mechanism of action in mammals, focusing on the different factors that could explain the worldwide distribution of the disease. Other indole diterpene alkaloids than lolitrem B that can be found in Epichloë infected ryegrass, and their tremorgenic properties, are presented in the last section of this review.

  19. Lolitrem B and Indole Diterpene Alkaloids Produced by Endophytic Fungi of the Genus Epichloë and Their Toxic Effects in Livestock

    Directory of Open Access Journals (Sweden)

    Guerre Philippe

    2016-02-01

    Full Text Available Different group of alkaloids are produced during the symbiotic development of fungal endophytes of the genus Epichloë in grass. The structure and toxicity of the compounds vary considerably in mammalian herbivores and in crop pests. Alkaloids of the indole-diterpene group, of which lolitrem B is the most toxic, were first characterized in endophyte-infected perennial ryegrass, and are responsible for “ryegrass staggers.” Ergot alkaloids, of which ergovaline is the most abundant ergopeptide alkaloid produced, are also found in ryegrass, but generally at a lower rate than lolitrem B. Other alkaloids such as lolines and peramine are toxic for crop pests but have weak toxicological properties in mammals. The purpose of this review is to present indole-diterpene alkaloids produced in endophyte infected ryegrass from the first characterization of ryegrass staggers to the determination of the toxicokinetics of lolitrem B and of their mechanism of action in mammals, focusing on the different factors that could explain the worldwide distribution of the disease. Other indole diterpene alkaloids than lolitrem B that can be found in Epichloë infected ryegrass, and their tremorgenic properties, are presented in the last section of this review.

  20. Nephrotoxicity evaluation of a new cembrane diterpene from Euphorbiae pekinensis Radix with HEK 293T cells and the toxicokinetics study in rats using a sensitive and reliable UFLC-MS/MS.

    Science.gov (United States)

    Zhang, Yuanyuan; Liu, Ziying; Zhang, Ruowen; Hou, Pengyi; Bi, Kaishun; Chen, Xiaohui

    2016-02-01

    (-)-(1S)-15-Hydroxy-18-carboxycembrene, the first cembrane-type diterpenoid found in the family Euphorbiaceae, isolated from Euphorbiae pekinensis Radix, was identified to be nephrotoxic using HEK 293T cells. Tests on cell morphology, cell viability and biochemical markers about oxidation stress were carried out using inverted microscope, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and commercial kits respectively, which proved the diterpene time- and dose-dependently decreased cells proliferation. Besides, a sensitive and robust UFLC-MS/MS method was developed and fully validated to investigate the in vivo behavior in rats of the diterpene after oral administration of Euphorbiae pekinensis Radix extracts at a dosage of 9g/kg. The method showed a good linearity in tested range (3-1500ng/mL) with acceptable accuracy and precision. The recovery of the diterpene was more than 85% and the matrix effect was within ±20%. The toxicokinetics parameters indicate the diterpene reached Cmax quickly and slowly eliminate. The study proved the newly found diterpene was one of the nephrotoxic substances of Euphorbiae pekinensis Radix and revealed its toxicokinetics behavior.

  1. Stevia rebaudiana ethanolic extract exerts better antioxidant properties and antiproliferative effects in tumour cells than its diterpene glycoside stevioside.

    Science.gov (United States)

    López, Víctor; Pérez, Sergio; Vinuesa, Arturo; Zorzetto, Christian; Abian, Olga

    2016-04-01

    Steviol glycosides are currently being used as natural sweeteners by the food industry and Stevia rebaudiana has long been used as a sweet plant in South America for patients suffering from diabetes. In this study, a Stevia rebaudiana ethanolic extract (SREE) was prepared, analysed and tested for antioxidant activity in terms of free radical scavenging properties and antiproliferative effects in cervix (HeLa), pancreatic (MiaPaCa-2) and colonic (HCT116) cancer cells. The antiproliferative mechanism was confirmed by testing the effects on cyclin D1-CDK4. Bioassays were also performed for the diterpene glycoside stevioside. Our results demonstrate that the extract acts as an antioxidant being able to scavenge free radicals, but this activity was not due to stevioside. The extract also induced cell death in the three cell lines, being more active against cervix cancer cells (HeLa); however, the concentration of stevioside needed to produce antiproliferative effects was higher than the amount of steviol glycosides found in a lower dose of extract inducing cell death. In addition, the extract clearly inhibited CDK4 whereas stevioside did not, concluding that the antiproliferative activity of stevia may be due to inhibition of cyclin-dependent kinases performed by other compounds of the extract.

  2. Quantitative Variation of Flavonoids and Diterpenes in Leaves and Stems of Cistus ladanifer L. at Different Ages

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    Cristina Valares Masa

    2016-02-01

    Full Text Available The compounds derived from secondary metabolism in plants perform a variety of ecological functions, providing the plant with resistance to biotic and abiotic factors. The basal levels of these metabolites for each organ, tissue or cell type depend on the development stage of the plant and they may be modified as a response to biotic and/or abiotic stress. As a consequence, the resistance state of a plant may vary in space and time. The secondary metabolites of Cistus ladanifer have been quantified in leaves and stems throughout autumn, winter, spring and summer, and at different ages of the plant. This study shows that there are significant differences between young leaves, mature leaves and stems, and between individuals of different ages. Young leaves show significantly greater synthesis of flavonoids and diterpenes than mature leaves and stems, with a clear seasonal variation, and the differences between leaves at different growth stages and stems is maintained during the quantified seasons. With respect to age, specimens under one year of age secreted significantly lower amounts of compounds. The variation in the composition of secondary metabolites between different parts of the plant, the season and the variations in age may determine the interactions of Cistus ladanifer with the biotic and abiotic factors to which it is exposed.

  3. Quantitative Variation of Flavonoids and Diterpenes in Leaves and Stems of Cistus ladanifer L. at Different Ages.

    Science.gov (United States)

    Valares Masa, Cristina; Sosa Díaz, Teresa; Alías Gallego, Juan Carlos; Chaves Lobón, Natividad

    2016-02-27

    The compounds derived from secondary metabolism in plants perform a variety of ecological functions, providing the plant with resistance to biotic and abiotic factors. The basal levels of these metabolites for each organ, tissue or cell type depend on the development stage of the plant and they may be modified as a response to biotic and/or abiotic stress. As a consequence, the resistance state of a plant may vary in space and time. The secondary metabolites of Cistus ladanifer have been quantified in leaves and stems throughout autumn, winter, spring and summer, and at different ages of the plant. This study shows that there are significant differences between young leaves, mature leaves and stems, and between individuals of different ages. Young leaves show significantly greater synthesis of flavonoids and diterpenes than mature leaves and stems, with a clear seasonal variation, and the differences between leaves at different growth stages and stems is maintained during the quantified seasons. With respect to age, specimens under one year of age secreted significantly lower amounts of compounds. The variation in the composition of secondary metabolites between different parts of the plant, the season and the variations in age may determine the interactions of Cistus ladanifer with the biotic and abiotic factors to which it is exposed.

  4. 紫珠的松香烷型二萜成分研究%Abietane diterpenes from Callicarpa pedunculata

    Institute of Scientific and Technical Information of China (English)

    芦毅; 华燕

    2011-01-01

    Five abietane diterpenes were isolated from CHCl3-solved portion of EtOH extraction from Callicarpa pedunculara R. Brown. On the basis of spectroscopic analysis,these compounds were identified as dehydroabietic acid (1) ,7β-hydroxydehydroabietic acid (2) ,7α-hydroxydehydroabietic acid (3) ,7-oxodehydroabietic acid (4), and 6,8,11,13-abietatrien-18-oic acid (5). These compounds were isolated from this plant for the first time.%从紫殊全草的乙醇提取物的氯仿部分中分得5个松香烷型二萜,经光谱数据分别鉴定为脱氢枞酸(1)、7β-羟基-脱氢枞酸(2)、7α-羟基-脱氢枞酸(3)、7-羰基-脱氢枞酸(4)和18-羧基-6,8,11,13-松香烷酸.以上化台物均为首次从该植物中分得.

  5. Proliferation inhibition, cell cycle arrest and apoptosis induced in HL-60 cells by a natural diterpene ester from Daphne mucronata

    Science.gov (United States)

    Nouri, K.; Yazdanparast, R.

    2011-01-01

    Background and the purpose of the study Gnidilatimonoein (Gn), a new diterpene ester from Daphne mucronata, possesses strong anti-metastasis and anti-tumor activities. In this study, its apoptosis and differentiation capabilities were evaluated by using the leukemia HL-60 cell line. Material and methods Cell prolifaration inhibition was estimated by MTT assay. The occurrence of apoptosis was evaluated by EtBr/AO double staining technique, cell cycle analyses and detection of apoptotic cells by Annexin V-FITC and propodium iodide (PI). Differentiation of the cells was determined by NBT reduction assay and the expression of specific cell surface markers such as CD14 and CD11b, were analyzed by flow cytometry. Results The drug decreased the growth of the cells dose- and time-dependently and the IC50 was found to be 1.3 µM. Our data suggested that Gn induced both monocytic differentiation and apoptosis among HL-60 cells. In addition, cell cycle analyses showed an increase in G1 phase population by 24 hrs, which was gradually replaced by Sub-G1 cell population (apoptotic cells) by 72 hrs. Conclusion Based on these data, the Gn-treated HL-60 cells displayed differentiation-dependent apoptosis. Thus, Gn might be a good candidate for differentiation therapy of leukemia, pending full biological evaluation of the compound among the wide array of leukemia cells. PMID:22615651

  6. Molecular Breeding of a Fungus Producing a Precursor Diterpene Suitable for Semi-Synthesis by Dissection of the Biosynthetic Machinery

    Science.gov (United States)

    Noike, Motoyoshi; Ono, Yusuke; Araki, Yuji; Tanio, Ryo; Higuchi, Yusuke; Nitta, Hajime; Hamano, Yoshimitsu; Toyomasu, Tomonobu; Sassa, Takeshi; Kato, Nobuo; Dairi, Tohru

    2012-01-01

    Many clinically useful pharmaceuticals are semi-synthesized from natural products produced by actinobacteria and fungi. The synthetic protocols usually contain many complicated reaction steps and thereby result in low yields and high costs. It is therefore important to breed microorganisms that produce a compound most suitable for chemical synthesis. For a long time, desirable mutants have been obtained by random mutagenesis and mass screening. However, these mutants sometimes show unfavorable phenotypes such as low viability and low productivity of the desired compound. Fusicoccin (FC) A is a diterpene glucoside produced by the fungus Phomopsis amygdali. Both FC and the structurally-related cotylenin A (CN) have phytohormone-like activity. However, only CN exhibits anti-cancer activity. Since the CN producer lost its ability to proliferate during preservation, a study on the relationship between structure and activity was carried out, and elimination of the hydroxyl group at position 12 of FC was essential to mimic the CN-like activity. Based on detailed dissection of the biosynthetic machinery, we constructed a mutant producing a compound without a hydroxyl group at position 12 by gene-disruption. The mutant produced this compound as a sole metabolite, which can be easily and efficiently converted into an anti-cancer drug, and its productivity was equivalent to the sum of FC-related compounds produced by the parental strain. Our strategy would be applicable to development of pharmaceuticals that are semi-synthesized from fungal metabolites. PMID:22870285

  7. Molecular breeding of a fungus producing a precursor diterpene suitable for semi-synthesis by dissection of the biosynthetic machinery.

    Directory of Open Access Journals (Sweden)

    Motoyoshi Noike

    Full Text Available Many clinically useful pharmaceuticals are semi-synthesized from natural products produced by actinobacteria and fungi. The synthetic protocols usually contain many complicated reaction steps and thereby result in low yields and high costs. It is therefore important to breed microorganisms that produce a compound most suitable for chemical synthesis. For a long time, desirable mutants have been obtained by random mutagenesis and mass screening. However, these mutants sometimes show unfavorable phenotypes such as low viability and low productivity of the desired compound. Fusicoccin (FC A is a diterpene glucoside produced by the fungus Phomopsis amygdali. Both FC and the structurally-related cotylenin A (CN have phytohormone-like activity. However, only CN exhibits anti-cancer activity. Since the CN producer lost its ability to proliferate during preservation, a study on the relationship between structure and activity was carried out, and elimination of the hydroxyl group at position 12 of FC was essential to mimic the CN-like activity. Based on detailed dissection of the biosynthetic machinery, we constructed a mutant producing a compound without a hydroxyl group at position 12 by gene-disruption. The mutant produced this compound as a sole metabolite, which can be easily and efficiently converted into an anti-cancer drug, and its productivity was equivalent to the sum of FC-related compounds produced by the parental strain. Our strategy would be applicable to development of pharmaceuticals that are semi-synthesized from fungal metabolites.

  8. Biflora-4,10(19),15-triene: a new diterpene from a termite soldier (isoptera termitidae termitinae)

    Energy Technology Data Exchange (ETDEWEB)

    Wiemer, D.F. (Cornell Univ., Ithaca, NY); Meinwald, J.; Prestwich, G.D.; Solheim, B.A.; Clardy, J.

    1980-01-04

    This communication describes spectrometric, chemical, and x-ray diffraction studies leading to the assignment of the structure to this diterpene. The name assigned is biflora-4,10(19),15-triene on the basis of its skeletal relationship to the violet-red, o-quinonoid antibiotic biflorin, a constituent of the Brazilian medicinal plant Capraria biflora L. The triene was isolated from a hexane extract. The mass spectra and NMR spectra were measured. The compound was esterified with p-bromobenzoyl chloride. The p-bromobenzoate crystallized in the monoclinic crystal system. Lattice parameters were measured. Intensity data were collected on a fully automated four-circle diffractometer. 1666 unique reflections were judged observed after correction for Lorentz, polarization, and background effects. The bromine position was deduced from the Patterson synthesis with complications. Careful selection of atoms finally yielded the correct structure. Full-matrix least-square refinements converged to the current minimum crystallographic residual of 0.090. A computer-generated perspective drawing of the final x-ray model is shown. 1 figure.

  9. Bioguided discovery and pharmacophore modeling of the mycotoxic indole diterpene alkaloids penitrems as breast cancer proliferation, migration, and invasion inhibitors

    Science.gov (United States)

    Sallam, Asmaa A.; Houssen, Wael E.; Gissendanner, Chris R.; Orabi, Khaled Y.; Foudah, Ahmed I.; El Sayed, Khalid A.

    2013-01-01

    Marine-derived fungi have proven to be important sources of bioactive natural organohalides. The genus Penicillium is recognized as a rich source of chemically diverse bioactive secondary metabolites. This study reports the fermentation, isolation and identification of a marine-derived Penicillium species. Bioassay-guided fractionation afforded the indole diterpene alkaloids penitrems A, B, D, E and F as well as paspaline and emnidole SB (1–7). Supplementing the fermentation broth of the growing fungus with KBr afforded the new 6-bromopenitrem B (8) and the known 6-bromopenitrem E (9). These compounds showed good antiproliferative, antimigratory and anti-invasive properties against human breast cancer cells. Penitrem B also showed a good activity profile in the NCI-60 DTP human tumor cell line screen. The nematode Caenorhabditis elegans was used to assess the BK channel inhibitory activity and toxicity of select compounds. A pharmacophore model was generated to explain the structural relationships of 1–9 with respect to their antiproliferative activity against the breast cancer MCF-7 cells. The structurally less complex biosynthetic precursors, paspaline (6) and emindole SB (7), were identified as potential hits suitable for future studies. PMID:24273638

  10. Structure-activity relationships for the irreversible blockade of nicotinic receptor agonist sites by lophotoxin and congeneric diterpene lactones

    Energy Technology Data Exchange (ETDEWEB)

    Culver, P.; Burch, M.; Potenza, C.; Wasserman, L.; Fenical, W.; Taylor, P.

    1985-11-01

    Lophotoxin, a diterpene lactone paralytic toxin from gorgonian corals of the genus Lophogorgia, inhibits ( SVI)-alpha-toxin binding to surface nicotinic receptors of BC3H-1 cells by irreversible occupation of the primary agonist sites. In contrast, receptor-bearing membrane fragments or detergent-solubilized receptors prepared from BC3H-1 cells are not susceptible to lophotoxin block. Thus, lophotoxin inhibition requires intact cells. However, when intact cells were incubated with lophotoxin, subsequent membrane-fragment preparation or detergent solubilization of the receptors did not diminish lophotoxin occupation of ( SVI)-alpha-toxin-binding sites, indicating that lophotoxin binds very tightly to nicotinic receptors. These studies further demonstrate that both surface and nonsurface nicotinic receptors of BC3H-1 cells are susceptible to irreversible occupation by lophotoxin, indicating that the lipophilic toxin freely permeates intact cells. The authors also examined several structural analogs of lophotoxin, one of which was equipotent with lophotoxin for inhibition of ( SVI)-alpha-toxin binding to intact cells and, notably, also blocked alpha-toxin binding to detergent-extracted receptor.

  11. Structure-based drug design studies of the interactions of ent-kaurane diterpenes derived from Wedelia paludosa with the Plasmodium falciparum sarco/endoplasmic reticulum Ca2+-ATPase PfATP6

    Directory of Open Access Journals (Sweden)

    Daniel Silqueira Martins Guimarães

    2015-04-01

    Full Text Available Malaria is responsible for more deaths around the world than any other parasitic disease. Due to the emergence of strains that are resistant to the current chemotherapeutic antimalarial arsenal, the search for new antimalarial drugs remains urgent though hampered by a lack of knowledge regarding the molecular mechanisms of artemisinin resistance. Semisynthetic compounds derived from diterpenes from the medicinal plant Wedelia paludosa were tested in silico against the Plasmodium falciparum Ca2+-ATPase, PfATP6. This protein was constructed by comparative modelling using the three-dimensional structure of a homologous protein, 1IWO, as a scaffold. Compound 21 showed the best docking scores, indicating a better interaction with PfATP6 than that of thapsigargin, the natural inhibitor. Inhibition of PfATP6 by diterpene compounds could promote a change in calcium homeostasis, leading to parasite death. These data suggest PfATP6 as a potential target for the antimalarial ent-kaurane diterpenes.

  12. O desamparo aprendido revisitado: estudos com animais

    Directory of Open Access Journals (Sweden)

    Maria Helena Leite Hunziker

    Full Text Available O desamparo aprendido tem sido definido como a dificuldade de aprendizagem apresentada por indivíduos que tiveram experiência prévia com estímulos aversivos incontroláveis. O objetivo deste trabalho é fazer uma revisão crítica dos estudos sobre o desamparo aprendido, com animais. Nessa análise, são considerados aspectos conceituais e metodológicos dos estudos em questão e as interpretações teóricas sobre esse efeito comportamental. Aborda-se a evolução histórica desses estudos, bem como alguns aspectos controversos das publicações que se acumularam ao longo de quatro décadas de pesquisa. A associação do desamparo aprendido com a depressão clínica é analisada criticamente, destacando-se a necessidade de maior rigor metodológico e conceitual nos estudos da área.

  13. Gastroprotective effect of barbatusin and 3-beta-hydroxy-3-deoxibarbatusin, quinonoid diterpenes isolated from Plectranthus grandis, in ethanol-induced gastric lesions in mice.

    Science.gov (United States)

    Rodrigues, Patrícia de Araújo; de Morais, Selene Maia; de Souza, Carolina Melo; Silva, Ana Raquel Araújo; de Andrade, Geanne Matos; Silva, Maria Goretti Vasconcelos; Albuquerque, Roberto Lima; Rao, Vietla Satyanarayana; Santos, Flávia Almeida

    2010-02-17

    Validate the popular use of Plectranthus grandis in gastric disorders through the active components. Isolation of barbatusin (BB) and 3beta-hydroxy-3-deoxibarbatusin (BBOH), diterpenes from Plectranthus grandis, and evaluation of their gastroprotective effect and possible mechanisms of action. Isolation and chemical characterization of diterpenes from Plectranthus grandis by chromatographic and spectroscopic methods and evaluation of gastroprotective action of the diterpenes through ethanol-induced gastric injury in mice model. It was evaluated the effect of capsazepine, indomethacin and the role of nitric oxide and K(ATP-) channels on the gastroprotective effect of BBOH and BB. Additionally it was measured the concentrations of gastric mucus, non-proteic-sulfhydryl groups and total thiobarbituric acid-reactive substances. Orally administered BBOH and BB at doses of 5 and 10mg/kg, markedly reduced the gastric lesions by 59 and 96%, and 32 and 76%, respectively, with superior results as compared to N-acetylcysteine (150 mg/kg, i.p.), reference compound that caused 85% lesion suppression. Although BBOH presented a higher gastroprotection than BB they act by similar mechanisms in relation to N-acetylcysteine, and prevent the depletion of gastric mucus, gastric mucosal non-proteic-sulfhydryl groups as well as the increase in thiobarbituric acid-reactive species. Moreover, the gastroprotective effect of BB was effectively blocked in mice pretreated with TRPV1 antagonist capsazepine, by the non-selective cyclooxygenase inhibitor indomethacin, or by the nitric oxide synthase inhibitor L-NAME but not by K(+)(ATP) channel inhibitor glibenclamide. In contrast, the gastroprotective effect of BBOH was blocked only by indomethacin and glibenclamide pretreatments. The protective role for BBOH and BB affording gastroprotection against gastric damage induced by ethanol indicates that these compounds contribute for the activity of Plectranthus species. The different modes of action

  14. Pharmacokinetics and tissue distribution of ginkgolide A, ginkgolide B, and ginkgolide K after intravenous infusion of ginkgo diterpene lactones in a rat model.

    Science.gov (United States)

    Wang, Shuyao; Ouyang, Bingchen; Aa, Jiye; Geng, Jianliang; Fei, Fei; Wang, Pei; Wang, Jiankun; Peng, Ying; Geng, Ting; Li, Yanjing; Huang, Wenzhe; Wang, Zhenzhong; Xiao, Wei; Wang, Guangji

    2016-07-15

    Ginkgo diterpene lactones are compounds that are extracted from the Ginkgo biloba leaf and possess pharmacologic activities with neuroprotective effects. To address the poor bioavailability of ginkgo diterpene lactones, ginkgo diterpene lactone meglumine injection (GDLI) was formulated and is commercially available. In this study, a simple, sensitive and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for assessing the total amount and the amount of the prototype forms of ginkgolides A (GA), B (GB) and K (GK) in rat plasma and tissues. This method was used to calculate the concentrations of the hydrolysed carboxylic forms and assess the pharmacokinetics of the ginkgolides after intravenous (i.v.) GDLI administration in rats. Generally, all three ginkgolide forms showed dose-dependent plasma concentrations, and no obvious differences in pharmacokinetic parameters, i.e., area under the curve (AUC) of plasma concentration versus time and half-life, were observed after GDLI administration on 7 consecutive days. These ginkgolides primarily existed in the carboxylic form in the plasma, and the systemic concentrations of the carboxylic forms of GA and GB were 11- to 17- and 3- to 4-fold higher than those of their prototype forms, respectively. In contrast, dramatically increased levels of the GA and GB prototype lactones were detected in the liver and heart. GA, GB, and GK were extensively distributed in various organs/tissues; the highest levels were found in the kidneys, liver, and intestine, and the lowest levels were found in the brain. These data suggest that ginkgolides have difficulty crossing the blood-brain barrier and that their targets for protecting against cerebral ischaemia are located outside the central system.

  15. Two Novel Diterpenes Isolated from the Flower Buds of Daphne genkwa%芫花花蕾中的两个新二萜

    Institute of Scientific and Technical Information of China (English)

    夏素霞; 李玲芝; 李菲菲; 彭缨; 宋少江; 高品一; 唐思

    2011-01-01

    瑞香烷型二萜类化合物具有良好的抗肿瘤生理活性.为了进一步研究该类抗肿瘤活性成分,对富含瑞香烷型二萜的芫花花蕾进行化学成分研究,从中分离得到一个巴豆烷型二萜1和两个瑞香烷型二萜原酸酯2和3.应用多种波谱学方法(1H NMR,13C NMR,HSQC,HMBC,NOESY,ESI-MS,HR-ESI-MS等),确定了3个化合物的结构,化合物1和2为新化合物,化合物1被鉴定为12-O-(2'E,4'E-decadienoyl)-4-hydroxyphorbol-13-acetyl.化合物2被鉴定为化合物3(芫花酯乙)的12位酰基烯烃链的顺式异构体,命名为异芫花酯乙.%Daphnane-type diterpenes showed potent anti-tumour activities,the flower buds of Daphne genkwa which was rich in daphnane-type diterpenes were subject to phytochemical investigation resulting in the isolation of two novel diterpenes(1,2) and a known diterpene 3,they were elucidated as 12-O-(2'E,4'E-decadienoyl)-4-hydroxyphorbol-13-acetyl(1),isoyuanhuadine(2) and yuanhuadine(3) by spectroscopic data,including 1H NMR,13C NMR,HSQC,HMBC,NOESY,ESI-MS and HR-ESI-MS.The new compound isoyanhuadine is yuanhuadine’s cis-isomer.

  16. A Synthetic Butenolide Diterpene is now a Natural Product Isolated from Metaporana sericosepala, a Plant from the Madagascar Dry Forest[1a

    Science.gov (United States)

    Presley, Christopher C.; Rakotondraibe, L. Harinantenaina; Brodie, Peggy J.; Callmander, Martin W.; Randrianaivo, Richard; Rasamison, Vincent E.; Rakotobe, Etienne; Kingston, David G. I.

    2015-01-01

    Antiproliferative bioassay-guided fractionation of the ethanolic extract of the endemic Madagascan plant Metaporana sericosepala led to the first natural product isolation of a butenolide diterpene which was synthesized during an anti-inflammatory study in 1988. The structure of the compound was elucidated as 3-homofarnesyl-4-hydroxybutenolide (1) by analysis of its spectroscopic data, including 1D- and 2D-NMR data and chemical evidence. The once synthetic compound can now also be considered as a natural product. Compound 1 had modest antiproliferative activity towards the A2780 ovarian cancer cell line with an IC50 value of 8 μM. PMID:26435765

  17. Biosynthesis of the psychotropic plant diterpene salvinorin A: Discovery and characterization of the Salvia divinorum clerodienyl diphosphate synthase.

    Science.gov (United States)

    Pelot, Kyle A; Mitchell, Rod; Kwon, Moonhyuk; Hagelthorn, David M; Wardman, Jacob F; Chiang, Angela; Bohlmann, Jörg; Ro, Dae-Kyun; Zerbe, Philipp

    2017-03-01

    Salvia divinorum commonly known as diviner's sage, is an ethnomedicinal plant of the mint family (Lamiaceae). Salvia divinorum is rich in clerodane-type diterpenoids, which accumulate predominantly in leaf glandular trichomes. The main bioactive metabolite, salvinorin A, is the first non-nitrogenous natural compound known to function as an opioid-receptor agonist, and is undergoing clinical trials for potential use in treating neuropsychiatric diseases and drug addictions. We report here the discovery and functional characterization of two S. divinorum diterpene synthases (diTPSs), the ent-copalyl diphosphate (ent-CPP) synthase SdCPS1, and the clerodienyl diphosphate (CLPP) synthase SdCPS2. Mining of leaf- and trichome-specific transcriptomes revealed five diTPSs, two of which are class II diTPSs (SdCPS1-2) and three are class I enzymes (SdKSL1-3). Of the class II diTPSs, transient expression in Nicotiana benthamiana identified SdCPS1 as an ent-CPP synthase, which is prevalent in roots and, together with SdKSL1, exhibits a possible dual role in general and specialized metabolism. In vivo co-expression and in vitro assays combined with nuclear magnetic resonance (NMR) analysis identified SdCPS2 as a CLPP synthase. A role of SdCPS2 in catalyzing the committed step in salvinorin A biosynthesis is supported by its biochemical function, trichome-specific expression and absence of additional class II diTPSs in S. divinorum. Structure-guided mutagenesis revealed four catalytic residues that enabled the re-programming of SdCPS2 activity to afford four distinct products, thus advancing our understanding of how neo-functionalization events have shaped the array of different class II diTPS functions in plants, and may promote synthetic biology platforms for a broader spectrum of diterpenoid bioproducts.

  18. Disruption of androgen and estrogen receptor activity in prostate cancer by a novel dietary diterpene carnosol: implications for chemoprevention

    Science.gov (United States)

    Johnson, Jeremy J.; Syed, Deeba N.; Suh, Yewseok; Heren, Chenelle R.; Saleem, Mohammad; Siddiqui, Imtiaz A.; Mukhtar, Hasan

    2010-01-01

    Emerging data is suggesting that estrogens, in addition to androgens, may also be contributing to the development of prostate cancer (PCa). In view of this notion agents that target estrogens, in addition to androgens, may be a novel approach for PCa chemoprevention and treatment. Thus, the identification and development of non-toxic dietary agents capable of disrupting androgen receptor (AR) in addition to estrogen receptor (ER) could be extremely useful in the management of PCa. Through molecular modeling we found carnosol, a dietary diterpene fits within the ligand binding domain of both AR and ER-α. Using a TR-FRET assay we found that carnosol interacts with both AR and ER-α and additional experiments confirmed that it functions as a receptor antagonist with no agonist effects. LNCaP, 22Rv1, and MCF7 cells treated with carnosol (20–40 µM) showed decreased protein expression of AR and ER-α. Oral administration of carnosol at 30 mg/kg five days weekly for 28 days to 22Rv1 PCa xenografted mice suppressed tumor growth by 36% (p = 0.028) and was associated with a decrease in serum PSA by 26% (p=0.0042). These properties make carnosol unique to any known anti-androgen or anti-estrogen investigated so far for the simultaneous disruption of AR and ER-α. We suggest that carnosol may be developed or chemically modified through more rigorous structure activity relationship studies for a new class of investigational agents - a dual AR/ER modulator. PMID:20736335

  19. Regiospecificities and Prenylation Mode Specificities of the Fungal Indole Diterpene Prenyltransferases AtmD and PaxD

    Science.gov (United States)

    Liu, Chengwei; Minami, Atsushi; Noike, Motoyoshi; Toshima, Hiroaki; Oikawa, Hideaki

    2013-01-01

    We recently reported the function of paxD, which is involved in the paxilline (compound 1) biosynthetic gene cluster in Penicillium paxilli. Recombinant PaxD catalyzed a stepwise regular-type diprenylation at the 21 and 22 positions of compound 1 with dimethylallyl diphosphate (DMAPP) as the prenyl donor. In this study, atmD, which is located in the aflatrem (compound 2) biosynthetic gene cluster in Aspergillus flavus and encodes an enzyme with 32% amino acid identity to PaxD, was characterized using recombinant enzyme. When compound 1 and DMAPP were used as substrates, two major products and a trace of minor product were formed. The structures of the two major products were determined to be reversely monoprenylated compound 1 at either the 20 or 21 position. Because compound 2 and β-aflatrem (compound 3), both of which are compound 1-related compounds produced by A. flavus, have the same prenyl moiety at the 20 and 21 position, respectively, AtmD should catalyze the prenylation in compound 2 and 3 biosynthesis. More importantly and surprisingly, AtmD accepted paspaline (compound 4), which is an intermediate of compound 1 biosynthesis that has a structure similar to that of compound 1, and catalyzed a regular monoprenylation of compound 4 at either the 21 or 22 position, though the reverse prenylation was observed with compound 1. This suggests that fungal indole diterpene prenyltransferases have the potential to alter their position and regular/reverse specificities for prenylation and could be applicable for the synthesis of industrially useful compounds. PMID:24038699

  20. Evident and latent plasticity across the rice diterpene synthase family with potential implications for the evolution of diterpenoid metabolism in the cereals

    Science.gov (United States)

    Morrone, Dana; Hillwig, Matthew L.; Mead, Matthew E.; Lowry, Luke; Fulton, D. Bruce; Peters, Reuben J.

    2013-01-01

    SYNOPSIS The evolution of natural products biosynthetic pathways can be envisioned to occur via a number of mechanisms. Here we provide evidence that latent plasticity plays a role in such metabolic evolution. In particular, rice (Oryza sativa) produces both ent- and syn-copalyl diphosphate (CPP), which are substrates for downstream diterpene synthases. Here we report that several members of this enzymatic family exhibit dual reactivity with some pairing of ent-, syn-, or normal CPP stereochemistry. Evident plasticity was observed, as a previously reported ent-sandaracopimaradiene synthase also converts syn-CPP to syn-labda-8(17),12E,14-triene, which can be found in planta. Notably, normal CPP is not naturally found in rice. Thus, the presence of diterpene synthases that react with this non-native metabolite reveals latent enzymatic/metabolic plasticity, providing biochemical capacity for utilization of such a novel substrate (i.e., normal CPP) that may arise during evolution, the implications of which are discussed. PMID:21323642

  1. Verwaarloosbare hoeveelheden cholesterolverhogende diterpenen in koffie gezet met de koffiepadzetmethode bij vergelijking met ongefilterde kookkoffie=Negligible amounts of cholesterol-raising diterpenes in coffee made with coffee pads in comparison with unfiltered coffee

    NARCIS (Netherlands)

    Boekschoten, M.V.; Cruchten, van S.T.J.; Kosmeijer-Schuil, J.G.; Katan, M.B.

    2006-01-01

    To determine the amounts of the serum-cholesterol raising diterpenes cafestol and kahweol in coffee made with coffee pads and the Senseo coffee machine as opposed to filtered and unfiltered coffee. DESIGN: Observational. METHOD: In five cities in the Netherlands coffee was purchased in three major

  2. Verwaarloosbare hoeveelheden cholesterolverhogende diterpenen in koffie gezet met de koffiepadzetmethode bij vergelijking met ongefilterde kookkoffie=Negligible amounts of cholesterol-raising diterpenes in coffee made with coffee pads in comparison with unfiltered coffee

    NARCIS (Netherlands)

    Boekschoten, M.V.; Cruchten, van S.T.J.; Kosmeijer-Schuil, J.G.; Katan, M.B.

    2006-01-01

    To determine the amounts of the serum-cholesterol raising diterpenes cafestol and kahweol in coffee made with coffee pads and the Senseo coffee machine as opposed to filtered and unfiltered coffee. DESIGN: Observational. METHOD: In five cities in the Netherlands coffee was purchased in three major s

  3. Effects of diterpene phenol extract of Rosmarinus Officinalis on TGFβ1 and mRNA expressions of its signaling pathway molecules in the lung tissue of pulmonary fibrosis rats

    Institute of Scientific and Technical Information of China (English)

    杨礼腾

    2013-01-01

    Objective To investigate the regulative mechanism of the diterpene phenol extract of Rosmarinus Officinalis (DERO) on the imbalance of collagen metabolism of the lung tissue in pulmonary fibrosis rats.Methods Fifty healthy Sprague-Dawley rats were randomly divided into normal saline group (NS) ,bleomycin-induced lung

  4. Formation of the Unusual Semivolatile Diterpene Rhizathalene by the Arabidopsis Class I Terpene Synthase TPS08 in the Root Stele Is Involved in Defense against Belowground Herbivory[W

    Science.gov (United States)

    Vaughan, Martha M.; Wang, Qiang; Webster, Francis X.; Kiemle, Dave; Hong, Young J.; Tantillo, Dean J.; Coates, Robert M.; Wray, Austin T.; Askew, Whitnee; O’Donnell, Christopher; Tokuhisa, James G.; Tholl, Dorothea

    2013-01-01

    Secondary metabolites are major constituents of plant defense against herbivore attack. Relatively little is known about the cell type–specific formation and antiherbivore activities of secondary compounds in roots despite the substantial impact of root herbivory on plant performance and fitness. Here, we describe the constitutive formation of semivolatile diterpenes called rhizathalenes by the class I terpene synthase (TPS) 08 in roots of Arabidopsis thaliana. The primary enzymatic product of TPS08, rhizathalene A, which is produced from the substrate all-trans geranylgeranyl diphosphate, represents a so far unidentified class of tricyclic diterpene carbon skeletons with an unusual tricyclic spiro-hydrindane structure. Protein targeting and administration of stable isotope precursors indicate that rhizathalenes are biosynthesized in root leucoplasts. TPS08 expression is largely localized to the root stele, suggesting a centric and gradual release of its diterpene products into the peripheral root cell layers. We demonstrate that roots of Arabidopsis tps08 mutant plants, grown aeroponically and in potting substrate, are more susceptible to herbivory by the opportunistic root herbivore fungus gnat (Bradysia spp) and suffer substantial removal of peripheral tissue at larval feeding sites. Our work provides evidence for the in vivo role of semivolatile diterpene metabolites as local antifeedants in belowground direct defense against root-feeding insects. PMID:23512856

  5. Comparison of the Pharmacological Effects of ShuXuening and Ginkgo Diterpene Lactone Meglumine Injection%舒血宁和银杏二萜内酯葡胺注射液药理作用比较研究

    Institute of Scientific and Technical Information of China (English)

    房利勤

    2015-01-01

    目的 比较舒血宁注射液和银杏二萜内酯葡胺注射液药理作用的区别,为临床使用提供参考.方法 选择性状相似的小鼠,给予不同剂量的舒血宁注射液和银杏二萜内酯葡胺注射液.结果 高剂量的舒血宁组和银杏二萜内酯葡胺组小鼠的单核-巨噬细胞碳粒廓清指数(K)和吞噬指数(α)均显著升高,均较对照组小鼠有显著性差异( P<0. 05),但银杏二萜内酯葡胺注射液的免疫效果更明显;高剂量的舒血宁和银杏二萜内酯葡胺注射液均显著降低小鼠的收缩压和舒张压,均较对照组具有显著性差异( P<0. 05),但银杏二萜内酯葡胺注射液的降压效果更明显;高剂量的舒血宁注射液和银杏二萜内酯葡胺注射液均能显著促进小鼠的睡眠,入睡比例和入睡时间均较对照组小鼠有显著性差异( P<0. 05),但银杏二萜内酯葡胺注射液组镇静效果更显著.结论 舒血宁注射液和银杏二萜内酯葡胺注射液均有较好的免疫、降血压、中枢镇静作用,但后者的药理活性更强.%Objective To compare the different pharmacological effects of Shuxuening injection and Ginkgo diterpene lactone meglumine Injection to provide suitable clinical reference. Methods Mice with similar traits were chosen and treated with different doses of Shux-uening injection and Ginkgo Diterpene Lactone Meglumine Injection. Results The high dose of Shuxuening Injection group and Ginkgo Diterpene Lactone Meglumine Injection group were significantly increased in mice monocyte-macrophage carbon particle clearance index K and index alpha of phagocytosis, and were lower than those in the control group with significant difference ( P < 0. 05 ) , but the im-mune effects of Ginkgo Diterpene Lactone Meglumine Injection group was more obvious; high dose of Shuxuening Injection group and Ginkgo Diterpene Lactone Meglumine Injection group were significantly decreased in mice systolic and diastolic blood pressure

  6. ESTUDO SOBRE O IMPACTO DOS PROCESSADORES HOSPEDEIROS

    Directory of Open Access Journals (Sweden)

    Alba S. B. Lopes

    2008-05-01

    Full Text Available Arquiteturas reconfiguráveis são dispositivos de hardware capazes de modificar sua estrutura de hardware de acordo com a aplicação a ser executada. Tais dispositivos surgiram como uma alternativa às soluções de hardware tradicionais na tentativa de equilibrar flexibilidade e desempenho. Uma das principais formas de utilização de arquiteturas reconfiguráveis corresponde a um bloco reconfigurável anexado a um processador hospedeiro. Esse processador é responsável por executar diversas tarefas críticas, incluindo o particionamento hardware/software e indicar o momento do bloco reconfigurável executar. Apesar de existirem diversas propostas de arquiteturas reconfiguráveis na literatura, pouco se estudou sobre o impacto do processador hospedeiro na arquitetura. Nesse contexto, esse artigo apresenta um estudo sobre o impacto do processador hospedeiro no desempenho da arquitetura reconfigurável híbrida. Como estudo de caso foi realizada uma avaliação sobre o uso dos processadores SPARC V8 da Sun Microsystems e Nios II da Altera como processadores hospedeiros da arquitetura reconfigurável híbrida RoSA. O objetivo deste estudo é realizar a comparação entre o desempenho desses processadores e identificar qual deles é o mais adequado para anexar a arquitetura em questão. Os resultados mostraram que foi possível alcançar até 47% de ganho de desempenho com o uso do processador Nios II em comparação ao SPARC V8, indicando o primeiro como mais adequado para a arquitetura RoSA. PALAVRAS-CHAVE: arquiteturas reconfiguráveis, processador hospedeiro, desempenho.

  7. 8,9-Dehydrohispanolone-15,16-lactol diterpene prevents LPS-triggered inflammatory responses by inhibiting endothelial activation.

    Science.gov (United States)

    Jiménez-García, Lidia; Través, Paqui G; López-Fontal, Raquel; Herranz, Sandra; Higueras, María Angeles; de Las Heras, Beatriz; Hortelano, Sonsoles; Luque, Alfonso

    2016-07-15

    Endothelial activation contributes to lung inflammatory disorders by inducing leucocyte recruitment to pulmonary parenchyma. Consequently, vascular-targeted therapies constitute promising strategies for the treatment of inflammatory pathologies. In the present study, we evaluated the effect of 8,9-dehydrohispanolone-15,16-lactol diterpene (DT) on lung endothelium during inflammation. Lung endothelial cells pre-treated with DT and activated with lipopolysaccharide (LPS) or tumour necrosis factor-α (TNF-α) exhibited reduced expression of the pro-inflammatory cytokines Cxcl10, Ccl5 and Cxcl1, whereas the anti-inflammatory molecules IL1r2 and IL-10 were induced. Consistent with this result, DT pre-treatment inhibited nuclear factor κB (NF-κB) nuclear translocation, by interfering with IκBα phosphorylation, and consequently NF-κB transcriptional activity in endothelium activated by LPS or TNF-α. Furthermore, DT, probably through p38 signalling, induced transcriptional activation of genes containing activator protein 1 (AP-1)-binding elements. Inhibition of p38 prevented IL1r2 mRNA expression in endothelium incubated with DT alone or in combination with LPS or TNF-α. Accordingly, conditioned medium (CM) from these cells failed to stimulate leucocytes as measured by a reduction in adhesive ability of the leucocyte cell line J774 to fibronectin (FN). Additionally, DT reduced the expression of the endothelial adhesion molecules E-selectin, vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) after activation. Similarly, expression of VCAM-1 and ICAM-1 molecules on the lung endothelial layer of C57/BL6 mice pre-treated with DT and challenged with LPS were unchanged. Finally, inhibition of vascular adhesion molecule expression by DT decreased the interaction of J774 cells with lung endothelial cells in an inflammatory environment. Our findings establish DT as a novel endothelial inhibitor for the treatment of inflammatory

  8. The vasorelaxant activity of marrubenol and marrubiin from Marrubium vulgare.

    Science.gov (United States)

    El Bardai, Sanae; Morel, Nicole; Wibo, Maurice; Fabre, Nicolas; Llabres, Gabriel; Lyoussi, Badiaa; Quetin-Leclercq, Joëlle

    2003-01-01

    Crude extracts of the aerial parts of Marrubium vulgare show a potent in vitro inhibition of KCl-induced contraction of rat aorta. Bio-guided fractionations, spectroscopic analysis and chemical derivatization revealed the furanic labdane diterpenes marrubenol and marrubiin as the most active compounds.

  9. A C-methylated resacetophenone from Cistus monspeliensis L.

    Science.gov (United States)

    Venditti, Alessandro; Bianco, Armandodoriano; Tomassini, Lamberto; Nicoletti, Marcello

    2014-06-01

    A new methylresacetophenone glucoside, named monspelioside, was isolated from Cistus monspeliensis, together with several diterpenes of labdane and clerodane types, a simple flavone and two methyl-inositols, one of them isolated for the first time from Cistus spp. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Phytochemistry and in vitro pharmacological activities of South African Vitex (Verbenaceae) species

    CSIR Research Space (South Africa)

    Nyiligira, E

    2008-10-01

    Full Text Available was purified to give a labdane diterpene as an inseparable epimeric mixture of 12S,16S/R-dihydroxy-ent-labda-7,13-dien-15,16-olide. Cirsimaritin was also isolated and identified from V. rehmannii. All the species, apart from V. zeyheri, exhibited scavenging...

  11. Antiparasitic, Nematicidal and Antifouling Constituents from Juniperus Berries

    Science.gov (United States)

    A bioassay-guided fractionation of Juniperus procera berries yielded antiparasitic, nematicidal and antifouling constituents, including a wide range of known abietane, pimarane and labdane diterpenes. Among these, abieta-7,13-diene (1) demonstrated in vitro antimalarial activity against Plasmodium f...

  12. Two New Diterpenoids from Hedychium forrestii

    Institute of Scientific and Technical Information of China (English)

    Qing ZHAO; Xin HONG; Yun Song WANG; Cheng ZOU; Xiao Jiang HAO

    2003-01-01

    Two new labdane diterpenes isolated from the rhizomes of Hedychium forrestii were determined by spectroscopic evidence to be labda-8(17), 11, 13-trien-7β-hydroxyl-15(16)-olide (1, hedyforrestin B) and labda-8(17), 11, 13-trien-7β,16-dihydroxyl-16(15)-olide (2, hedyforrestin C).

  13. Pine needle abortion in cattle update: Metabolite detection in sera and fetal fluids from abortion case samples

    Science.gov (United States)

    Cattle abortions associated with consumption of pine needles during late gestation are a serious poisonous plant problem in the Western US. Most cases of abortion have been associated with consumption of ponderosa pine (Pinus ponderosa) and the causative agent was identified as the labdane diterpen...

  14. Traditional preparation of Phaleria nisidai, a Palauan tea, reduces exposure to toxic daphnane-type diterpene esters while maintaining immunomodulatory activity.

    Science.gov (United States)

    Kulakowski, Daniel; Kitalong, Christopher; Negrin, Adam; Tadao, Van-Ray; Balick, Michael J; Kennelly, Edward J

    2015-09-15

    The leaves of Phaleria nisidai Kaneh. (Thymelaeaceae) are brewed into a tea commonly used as a tonic, strengthening beverage and immune enhancer in Palau, Micronesia. Recently, the leaves of P. nisidai have been shown to contain toxic daphnane diterpene esters which may pose a public health threat to Palauans. This project documents the use frequency, preparation and side effects of P. nisidai. The content of daphnane diterpene esters in aqueous and methanol extracts and infusions prepared by healers in Palau is compared to assess the risk of daphnane ingestion associated with traditional consumption. Quantitative results are correlated with an in vitro assessment of the immunomodulating activity of the extracts. Research participants, comprising traditional healers and laypeople, were interviewed concerning use patterns and side effects of P. nisidai. Several traditional healers prepared and provided boiled tea samples for chemical analysis. Leaves were collected and methanolic and aqueous extractions were prepared in the laboratory. Peripheral blood mononuclear cells (PBMCs) were cultured with various concentrations of methanol and aqueous leaf extracts and their output of IFNγ was measured using ELISA. Cell proliferation was also assessed using the MTT assay. The concentration of selected daphnane diterpene esters in healer-prepared infusions, lab methanol and lab aqueous extracts was quantified using ultraperformance liquid chromatography-mass spectrometry-triple quadrupole detection (UPLC-MS-TQD). Through structured interviews it was determined that P. nisidai tea was used frequently, with many participants drinking it daily. The reported side effects were mild, and with the exception of diarrhea (n=2), no side effect was mentioned more than once. Methanol extracts contained 4.0μg simplexin, 17.6μg acetoxyhuratoxin and 2.3μg huratoxin per g dry leaf material. In traditional water infusions provided by healers and in standardized lab-prepared aqueous

  15. Cholesterol-raising diterpenes in types of coffee commonly consumed in Singapore, Indonesia and India and associations with blood lipids: A survey and cross sectional study

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    Buchmann Sandra

    2011-05-01

    Full Text Available Abstract Background To measure the content of cholesterol-raising diterpenes in coffee sold at the retailer level in Singapore, Indonesia and India and to determine the relationship of coffee consumption with lipid levels in a population-based study in Singapore. Methods Survey and cross-sectional study in local coffee shops in Singapore, Indonesia and India to measure the diterpene content in coffee, and a population-based study in Singapore to examine the relationship of coffee consumption and blood lipid levels. Interviews and coffee samples (n = 27 were collected from coffee shops in Singapore, Indonesia and India. In addition, 3000 men and women who were Chinese, Malay, and Indian residents of Singapore participated in a cross-sectional study. Results and Discussion The traditional 'sock' method of coffee preparation used in Singapore resulted in cafestol concentrations comparable to European paper drip filtered coffee (mean 0.09 ± SD 0.064 mg/cup. This amount would result in negligible predicted increases in serum cholesterol and triglyceride concentrations. Similarly low amounts of cafestol were found in Indian 'filter' coffee that used a metal mesh filter (0.05 ± 0.05 mg/cup. Coffee samples from Indonesia using the 'sock' method (0.85 ± 0.41 mg/cup or a metal mesh filter (0.98 mg/cup contained higher amounts of cafestol comparable to espresso coffee. Unfiltered coffee from Indonesia contained an amount of cafestol (4.43 mg/cup similar to Scandinavian boiled, Turkish and French press coffee with substantial predicted increases in serum cholesterol (0.33 mmol/l and triglycerides (0.20 mmol/l concentrations for consumption of 5 cups per day. In the Singaporean population, higher coffee consumption was not substantially associated with serum lipid concentrations after adjustment for potential confounders [LDL-cholesterol: 3.07 (95% confidence interval 2.97-3.18 for Conclusions Based on the low levels of diterpenes found in traditionally

  16. Hydrogenation of the Exocyclic Olefinic Bond at C-16/C-17 Position of ent-Kaurane Diterpene Glycosides of Stevia rebaudiana Using Various Catalysts

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    Indra Prakash

    2013-07-01

    Full Text Available Catalytic hydrogenation of the exocyclic double bond present between C16 and C17 carbons of the four ent-kaurane diterpene glycosides namely rebaudioside A, rebaudioside B, rebaudioside C, and rebaudioside D isolated from Stevia rebaudiana has been carried out using Pt/C, Pd(OH2, Rh/C, Raney Ni, PtO2, and 5% Pd/BaCO3 to their corresponding dihydro derivatives with 17α and 17β methyl group isomers. Reactions were performed using the above-mentioned catalysts with the solvents methanol, water, and ethanol/water (8:2 under various conditions. Synthesis of reduced steviol glycosides was performed using straightforward chemistry and their structures were characterized on the basis of 1D and 2D NMR spectral data, including a comparison with reported spectral data.

  17. Hydrogenation of the Exocyclic Olefinic Bond at C-16/C-17 Position of ent-Kaurane Diterpene Glycosides of Stevia rebaudiana Using Various Catalysts

    Science.gov (United States)

    Chaturvedula, Venkata Sai Prakash; Prakash, Indra

    2013-01-01

    Catalytic hydrogenation of the exocyclic double bond present between C16 and C17 carbons of the four ent-kaurane diterpene glycosides namely rebaudioside A, rebaudioside B, rebaudioside C, and rebaudioside D isolated from Stevia rebaudiana has been carried out using Pt/C, Pd(OH)2, Rh/C, Raney Ni, PtO2, and 5% Pd/BaCO3 to their corresponding dihydro derivatives with 17α and 17β methyl group isomers. Reactions were performed using the above-mentioned catalysts with the solvents methanol, water, and ethanol/water (8:2) under various conditions. Synthesis of reduced steviol glycosides was performed using straightforward chemistry and their structures were characterized on the basis of 1D and 2D NMR spectral data, including a comparison with reported spectral data. PMID:23896597

  18. Structure activity relationship of phenolic diterpenes from Salvia officinalis as activators of the nuclear factor E2-related factor 2 pathway.

    Science.gov (United States)

    Fischedick, Justin T; Standiford, Miranda; Johnson, Delinda A; Johnson, Jeffrey A

    2013-05-01

    Nuclear factor E2-related factor 2 (Nrf2) is a transcription factor known to activate cytoprotective genes which may be useful in the treatment of neurodegenerative disease. In order to better understand the structure activity relationship of phenolic diterpenes from Salvia officinalis L., we isolated carnosic acid, carnosol, epirosmanol, rosmanol, 12-methoxy-carnosic acid, sageone, and carnosaldehyde using polyamide column, centrifugal partition chromatography, and semi-preparative high performance liquid chromatography. Isolated compounds were screened in vitro for their ability to active the Nrf2 and general cellular toxicity using mouse primary cortical cultures. All compounds except 12-methoxy-carnosic acid were able to activate the antioxidant response element. Furthermore both carnosol and carnoasldehyde were able to induce Nrf2-dependent gene expression as well as protect mouse primary cortical neuronal cultures from H(2)O(2) induced cell death.

  19. Structure-activity relationships for euphocharacins A-L, a new series of jatrophane diterpenes, as inhibitors of cancer cell P-glycoprotein.

    Science.gov (United States)

    Corea, Gabriella; Fattorusso, Ernesto; Lanzotti, Virginia; Motti, Riccardo; Simon, Pierre-Noël; Dumontet, Charles; Di Pietro, Attilio

    2004-07-01

    The Mediterranean spurge Euphorbia characias L. afforded twelve new diterpenes based on a jatrophane skeleton named euphocharacins A-L. Their chemical structures were elucidated by extensive nuclear magnetic resonance and mass spectrometry methods. Euphocharacins A-L were tested as inhibitors of the daunomycin-efflux activity of P-glycoprotein from cancer cells. The results were used to extend the structure-activity relationship established for this class of compounds, highlighting the positive effects of propyl and benzoyl groups at positions 3 and 9, respectively, and evidencing the negative effect of a free hydroxyl group at position 2. Among the tested compounds, euphocharacins C and I showed an activity higher than cyclosporin to inhibit Pgp-mediated daunomycin transport.

  20. Lathyrol diterpenes as modulators of P-glycoprotein dependent multidrug resistance: structure-activity relationship studies on Euphorbia factor L3 derivatives.

    Science.gov (United States)

    Jiao, Wei; Wan, Zhongmin; Chen, Shuang; Lu, Runhua; Chen, Xiaozhen; Fang, Dongmei; Wang, Jiufeng; Pu, Shengcai; Huang, Xin; Gao, Haixiang; Shao, Huawu

    2015-05-14

    Five series of 37 new acylate and epoxide derivatives (3-39) of Euphorbia factor L3, a lathyrol diterpene isolated from Euphorbia lathyris, were designed by modifying the hydroxyl moiety of C-3, C-5, or C-15. Chemoreversal effects of the acylates on multidrug resistance (MDR) were evaluated in breast cancer multidrug-resistant MCF-7/ADR cells that overexpress P-glycoprotein (P-gp). Eight derivatives exhibited greater chemoreversal ability than verapamil (VRP) against adriamycin (ADR) resistance. Compounds 19 and 25 exhibited 4.8 and 4.0 times, respectively, more effective reversal ability than VRP against ADR resistance. To determine the key characteristics of Euphorbia factor L3 derivatives that contribute to MDR reversal, we conducted a structure-activity relationship study of these compounds. The simulation studies indicated different possible mechanisms and revealed the important influence of hydrophobic interactions and hydrogen bonds in the flexible cavity of P-gp.

  1. Hydrogenation of the exocyclic olefinic bond at C-16/C-17 position of ent-kaurane diterpene glycosides of Stevia rebaudiana using various catalysts.

    Science.gov (United States)

    Chaturvedula, Venkata Sai Prakash; Prakash, Indra

    2013-07-26

    Catalytic hydrogenation of the exocyclic double bond present between C16 and C17 carbons of the four ent-kaurane diterpene glycosides namely rebaudioside A, rebaudioside B, rebaudioside C, and rebaudioside D isolated from Stevia rebaudiana has been carried out using Pt/C, Pd(OH)2, Rh/C, Raney Ni, PtO2, and 5% Pd/BaCO3 to their corresponding dihydro derivatives with 17α and 17β methyl group isomers. Reactions were performed using the above-mentioned catalysts with the solvents methanol, water, and ethanol/water (8:2) under various conditions. Synthesis of reduced steviol glycosides was performed using straightforward chemistry and their structures were characterized on the basis of 1D and 2D NMR spectral data, including a comparison with reported spectral data.

  2. Composition of Xylopia aethiopica (Dunal A. Rich essential oils from Cameroon and identification of a minor diterpene: ent-13-epi manoyl oxide

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    François Malaisse

    2007-01-01

    Full Text Available Xylopia aethiopica (Annonaceae essential oil was extracted from fruits collected in four localities in Cameroon, and analysed by GC/MS and GC/FID. More than sixty compounds were identified with 47.5–84.0% of monoterpenes hydrocarbon, mainly b-pinene and b-phellandrene+1,8-cineole, 6.5–12.9% of oxygenated monoterpenes, 13.8–30.4% of sesquiterpenes, and 0.4–0.6 % of a minor unidentified diterpene. Trials of purification by column chromatography, followed by GC/MS and NMR analysis led to the identification of ent-13-epi manoyl oxide which is reported for the first time as a minor component in X. aethiopica essential oil.

  3. 5-Episinuleptolide Acetate, a Norcembranoidal Diterpene from the Formosan Soft Coral Sinularia sp., Induces Leukemia Cell Apoptosis through Hsp90 Inhibition

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    Yao-Tsung Yeh

    2013-03-01

    Full Text Available 5-Episinuleptolide acetate (5EPA, a cytotoxic norcembranoidal diterpene recently identified from the Formosan soft coral Sinularia sp., exhibited potent activity against the K562, Molt 4 and HL 60 cancer cell lines. The antiproliferative assay, as well as the annexin V-FITC/propidium iodide (PI apoptotic assay, indicated that the HL 60 cell line is the most sensitive one towards 5EPA. This diterpenoid led to caspases -3, -8, and -9 activation as well as PARP cleavage. It also induced ROS generation, calcium accumulation and disruption of mitochondrial membrane potential. Additionally, the expression levels of Hsp90 protein and several client proteins were downregulated in response to 5EPA treatment. These results suggest that 5EPA’s cytotoxic effect on HL 60 cells may be attributed to the inhibition of Hsp90 as well as the induction of mitochondrial stress which finally results in apoptotic cell death.

  4. Estudo do teor de alicina em alho

    OpenAIRE

    Mendes, Patrícia Alexandra Pinto

    2008-01-01

    A alicina (dialil-tiosulfinato) é o componente biológico mais activo no alho com inúmeras aplicações a nível da saúde, já conhecida desde décadas. Existem actualmente diversos métodos publicados para a determinação do teor de alicina no alho por HPLC e espectrofotometria. No entanto, os resultados mostram discordância no teor de alicina medido pelos vários métodos. Este estudo tem por objectivo a determinação do teor de alicina por métodos de HPLC e espectrofotometria; e com...

  5. O estudo do comportamento verbal no Brasil

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    Adriana Pineiro Fidalgo

    Full Text Available O presente trabalho realizou uma revisão histórica de dissertações de mestrado e teses de doutorado brasileiras sobre comportamento verbal, com base na proposta Skinneriana (1957/1992, produzidas entre 1968 e 2012. Foram investigados: a tipo de trabalho (dissertação ou tese, b universidades em que os trabalhos foram defendidos, c orientadores, d linha de pesquisa (básica, aplicada ou histórico-conceitual, e metodologia (descritiva ou experimental, e f temas de investigação. No total, 177 dissertações e 53 teses sobre comportamento verbal foram identificadas. Os resultados indicam que o estudo do comportamento verbal, no Brasil, estabeleceu-se como programa de pesquisa e cresceu ao longo dos anos.

  6. Estudo forense do sémen

    OpenAIRE

    Vaz, Josiana A.; Chelas, S.; S Santos; Queirós, B.; Gonçalves, A.; Pereira, M.; Alves, Maria José

    2009-01-01

    O estudo dos fluidos seminais na cena do crime está directamente ligado a crimes de índole sexual, sendo este de importância vital aquando da reconstrução do acto do crime e na identificação do agressor. O sémen é segregado pelos órgãos reprodutores masculinos, sendo o suporte líquido dos espermatozóides. Segundo Pinheiro (2008), este vestígio pode ser encontrado em manchas no vestuário, lençóis, almofadas, móveis, chão, veículos, tapetes, entre outros. O sémen, antes de secar, possui um o...

  7. DADOS ESPECTROSCÓPICOS DE DITERPENOS LABDÂNICOS: UMA ANÁLISE TEÓRICA VIA RMN E DFT

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    Fabrine S. de Souza

    2015-05-01

    Full Text Available Labdane diterpenes exhibit important bioactivities such as cardiovascular effects in rats as well as effects in the treatment of autoimmune diseases and Alzheimer syndrome. Recently, the labdane diterpenes ent-13-epi-manoil oxide, ribenone and ribenol were isolated from Croton palanostigma. The computational method DFT/B3LYP/cc-pVDZ was used to optimize the structures of these diterpenes and to calculate infrared data. Chemical shifts (δH and δC of the minimum energy structures (local minimum were calculated and compared with the experimental data. Comparison of the NMR data by simple linear regression (SLR showed satisfactory statistical results with a correlation coefficient (R2 and predictive ability (Q2 of over 98%. The predicted NMR data were used to confirm the δH values that have not been published.

  8. Asymmetric Synthesis of Natural Macrocyclic Diterpene(+)-Methyl Cembra-1,3,7,11-tetraene-16-carboxynate%(+)-Methyl Cembra-1,3,7,11-tetraene-16-carboxynate的不对称合成研究

    Institute of Scientific and Technical Information of China (English)

    孙彬; 梅天胜; 刘佐胜; 李裕林; 李瀛; 彭立增

    2005-01-01

    The first asymmetric synthesis of (+)-methyl cembra-1,3,7,11-tetraene-16-carboxynate, a naturally occurring cembrane-type macrocyclic diterpene isolated from Sinularia mayi, was achieved via general approach by employing an intramolecular McMurry coupling and Sharpless asymmetric epoxidation as the key steps from readily available starting materials. The synthesis presented here verifies that the absolute configuration of compound 1 was assumed as 15R.

  9. Inhibition of the nuclear factor kappa B (NF-kappa B) pathway by tetracyclic kaurene diterpenes in macrophages. Specific effects on NF-kappa B-inducing kinase activity and on the coordinate activation of ERK and p38 MAPK.

    Science.gov (United States)

    Castrillo, A; de Las Heras, B; Hortelano, S; Rodriguez, B; Villar, A; Bosca, L

    2001-05-11

    The anti-inflammatory action of most terpenes has been explained in terms of the inhibition of nuclear factor kappaB (NF-kappaB) activity. Ent-kaurene diterpenes are intermediates of the synthesis of gibberellins and inhibit the expression of NO synthase-2 and the release of tumor necrosis factor-alpha in J774 macrophages challenged with lipopolysaccharide. These diterpenes inhibit NF-kappaB and IkappaB kinase (IKK) activation in vivo but failed to affect in vitro the function of NF-kappaB, the phosphorylation and targeting of IkappaBalpha, and the activity of IKK-2. Transient expression of NF-kappaB-inducing kinase (NIK) activated the IKK complex and NF-kappaB, a process that was inhibited by kaurenes, indicating that the inhibition of NIK was one of the targets of these diterpenes. These results show that kaurenes impair the inflammatory signaling by inhibiting NIK, a member of the MAPK kinase superfamily that interacts with tumor necrosis factor receptor-associated factors, and mediate the activation of NF-kappaB by these receptors. Moreover, kaurenes delayed the phosphorylation of p38, ERK1, and ERK2 MAPKs, but not that of JNK, in response to lipopolysaccharide treatment of J774 cells. The absence of a coordinate activation of MAPK and IKK might contribute to a deficient activation of NF-kappaB that is involved in the anti-inflammatory activity of these molecules.

  10. Structural elucidation of a new cembranoid diterpene from the Chinese soft coral Sarcophyton sp.%软珊瑚Sarcophyton sp.中一个新西松烷型二萜的结构解析

    Institute of Scientific and Technical Information of China (English)

    别玮; 邓志威; 徐岷涓; 林文翰

    2008-01-01

    对南海软珊瑚Sarcophyton sp.中的西松烷二萜类成份进行研究,从发现具有新颖结构的次生代谢产物.采用多种色谱手段进行分离纯化,通过理化性质和光谱分析确定化合物结构.分离得到1个新的西松烷二萜类化合物,命名为Sarcophyolide A(1),和一个已知西松烷二萜类化合物7α,8β-dihydroxydeepoxysarcophine(2).利用现代波谱确定新化合物的结构.%To investigate the eembranoid diterpenes from the soft coral Sarcophyton sp. collected in the South China Sea. Re-peatedly column chromatography was performed for the isolation and purification. The structures were elucidated on the basis of extensive spectral data (IR, MS, ID- and 2D-NMR) analysis by comparing with literature data. A new eembranoid diterpene namely sareophyolide A (1), together with 7a, 8β-dihydroxydeepoxysareophine (2) were isolated from this species. The cembra-noid diterpenes with γ-lactone could be chemotaxonomic markers ofSarcophyton sp.

  11. Blocking the L-type Ca2+ channel (Cav 1.2) is the key mechanism for the vascular relaxing effect of Pterodon spp. and its isolated diterpene methyl-6α-acetoxy-7β-hydroxyvouacapan-17β-oate.

    Science.gov (United States)

    de Fátima Reis, Carolina; de Andrade, Daniela Medeiros Lobo; Junior Neves, Bruno; de Almeida Ribeiro Oliveira, Leandra; Pinho, José Felippe; da Silva, Leidiane Pinha; Dos Santos Cruz, Jader; Bara, Maria Teresa Freitas; Andrade, Carolina Horta; Rocha, Matheus Lavorenti

    2015-10-01

    Pterodon spp. Vogel (Fabaceae), popularly known as "sucupira", has ethnopharmacological application which is described as having antispasmodic and relaxant effects. Hence, it was hypothesized that sucupira oil-resin (SOR) could induce smooth muscle relaxation. So, this study investigated the mechanisms involved in the vasorelaxant effect of SOR and its isolated diterpene (methyl-6α-acetoxy-7β-hydroxyvouacapan-17β-oate). Vascular reactivity experiments were performed using rat aortic rings (n=5-8) with (E+) or without endothelium (E-) in an isolated bath organ. The SOR (0-56 μg/mL) relaxed phenylephrine (E+: 86.7±7.1%; E-: 92.3±4.7%) and KCl contracted rings (E-: 97.1±2.8%). In the same way, diterpene (0-48 μg/mL) also relaxed phenylephrine (E+: 94.5±3.6%; E-: 92.2±3.4%) and KCl contracted rings (E-: 99.7±0.2%). The pre-incubation of arterial rings with cyclopiazonic acid (reticular Ca2+-ATPase inhibitor), tetraethylammonium (K+ channels blocker) or MDL-12,330A (adenylyl cyclesinhibitor) did not modify either SOR- or diterpeneinduced vasorelaxation. However, ODQ (guanylyl cyclase inhibitor) impaired only diterpene-induced vasorelaxation. SOR and diterpene significantly reduced CaCl2-induced contraction stimulated by Bay K8644 (1 μM), phenylephrine (0.1 μM) or KCl solution (40 mM). Computational molecular docking studies demonstrated that the vasodilator effect of diterpene relies on blocking the Cav 1.2 channel, and patch clamp results showed that diterpene substantially decreased the ionic current through Cav 1.2 in freshly dissociated vascular smooth muscle cells. These findings suggest that SOR and its isolated diterpene induce endothelium-independent vascular relaxation by blocking the L-type Ca2+ channel (Cav 1.2).

  12. ESTUDO PROSPECTIVO SOBRE ENCAPSULAMENTO DE COMPOSTOS BIOATIVOS

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    Larissa Assunção

    2014-12-01

    Full Text Available Com o advento da nanotecnologia, houve o desenvolvimento de técnicas envolvendo processamento, fabricação e aplicação de estruturas, com forma e tamanho controlados, sendo que na indústria de alimentos, estas formulações, tanto na faixa micrométrica como nanométrica, vêm sendo utilizadas com o objetivo de favorecer o produto final. Tendo em vista a importância tecnológica dessa técnica e do grande interesse nacional e internacional, principalmente no ramo da indústria alimentícia, sobre os benefícios do encapsulamento, o presente estudo prospectivo teve como objetivo avaliar o panorama mundial relativo ao tema em questão, correlacionando-o com os documentos de patentes depositados. Para tanto, realizou-se uma pesquisa das patentes na base de dados europeia Espacenet®, utilizando os descritores do tema em estudo associados aos códigos de classificação. As informações encontradas foram compiladas em gráficos e discutidas. Pôde-se observar que o maior número de patentes referiu-se ao código A23L1/00 - alimentos ou produtos alimentícios: a sua preparação ou tratamento e houve um crescimento exponencial em relação ao depósito de patentes nos últimos 10 anos, sendo os Estados Unidos o país líder em número de patentes depositadas (60%. A Firmenich e CIE lidera o ranking com mais patentes depositadas. Em relação à área de atuação, predomina a indústria alimentícia (69%. No que diz respeito ao Brasil, o país aparece com apenas 1 patente detentora desta tecnologia, sendo um número bastante reduzido o que mostra a falta de incentivo neste setor. Diante dos resultados obtidos, concluiu-se que, apesar de ser uma tecnologia recente, observa-se uma tendência ao crescimento de depósito de patentes.

  13. RUDOLF LABAN, A COREOLOGIA E OS ESTUDOS COREOLÓGICOS

    OpenAIRE

    Mota, Julio

    2012-01-01

    Este trabalho visa examinar como a necessidade de Laban conhecer as leis que regem o movimento humano o levou ao desenvolvimento de um método científico de estudo a que ele chamou de Coreologia. E de como a necessidade de aplicar o método coreológico, de maneira específica à Dança, conduziu ao surgimento de uma área específica de estudos, denominada Estudos Coreológicos. Este processo é aqui analisado através de um breve resumo biográfico de Laban. Essa biografia mostra como as in...

  14. Estudo retrospectivo de latrodectismo na Bahia, Brasil

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    Rejâne Maria Lira-da-Silva

    1995-09-01

    Full Text Available O trabalho apresenta um estudo retrospectivo de setenta e sete casos de latrodectismo no Estado da Bahia, Brasil, de agosto de 1980 a julho de 1990. Os dados foram levantados nos livros de registro e arquivo de fichas do CIAVE. O agente etiológico em 28% dos acidentes aracnídeos foi a espécie L. curacaviensis e a maior incidência foi registrada no meio urbano (57%, em indivíduos do sexo masculino (70% e faixa etária de 10 a 29 anos (58%. Os principais sinais locais foram dor (56%, pápula eritematosa (21% e edema discreto (17%, e os sistêmicos foram dor em membros inferiores (29%, tremores e contraturas (29%, sudorese (28% parestesia em membros (21% e dor abdominal (17%. O tratamento foi sintomático em 67% dos casos e específico em 21%. O tempo de permanência hospitalar após o uso do soro antilatrodectus foi menor que 24 horas em 64% dos casos.

  15. Habilidades Sociais em Alcoolistas: Um Estudo Comparativo

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    Flaviane Bevilaqua Felicissimo

    Full Text Available RESUMO O presente estudo objetivou comparar o repertório de habilidades sociais (HS de dependentes e não dependentes de álcool a fim de verificar se um menor repertório de HS constitui uma característica dessa população. Foram entrevistados 123 alcoolistas e 114 usuários de serviços de saúde com baixo ou nenhum consumo de álcool, utilizando questionário sociodemográfico, Mini International Neuropsychiatric Interview e o Inventário de Habilidades Sociais. Os dados foram analisados quantitativamente e indicaram uma diferença significativa no repertório de HS, especificamente no fator de autocontrole da agressividade, indicando um pior desempenho dessa habilidade em alcoolistas. Os resultados sugerem que a avaliação dessa característica deva ser considerada no tratamento do alcoolismo e associada ao plano terapêutico.

  16. Paralisia periodica familiar: estudo de oito casos

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    José Lamartine De Assis

    1980-12-01

    Full Text Available Foram estudados oito pacientes com Paralisia Periódica Familiar sob os aspectos clínico, hidro-eletrolítico, eletrofisiológico, histológico e terapêutico. Houve predomínio significante em pacientes do sexo masculino. A idade média do início da moléstia foi de 15 anos. As manifestações clínicas foram concor-dantes com as referidas na literatura. Todos os pacientes tinham a forma hipocalêmica exceto um que evoluiu com hipercalemia. Alguns pacientes (21,4% tinham natremias no limite superior da normalidade enquanto em um número igual de pacientes as natremias estavam elevadas, o que sugeria liberação de aldosterona. Os achados eletrofisiológicos foram concordantes com os da literatura. O estudo histológico de músculos realizado em cinco pacientes foi normal em 30% deles e, naqueles com crises repetidas e freqüentes, foram encontradas atrofias de fibras musculares. Foram feitos tratamentos preventivos, e nos pacientes com hipocalemia foram administradas doses suplementares de potássio em solução aquosa de 10-25% diariamente.

  17. Cholesterol-raising diterpenes in types of coffee commonly consumed in Singapore, Indonesia and India and associations with blood lipids: A survey and cross sectional study

    Science.gov (United States)

    2011-01-01

    Background To measure the content of cholesterol-raising diterpenes in coffee sold at the retailer level in Singapore, Indonesia and India and to determine the relationship of coffee consumption with lipid levels in a population-based study in Singapore. Methods Survey and cross-sectional study in local coffee shops in Singapore, Indonesia and India to measure the diterpene content in coffee, and a population-based study in Singapore to examine the relationship of coffee consumption and blood lipid levels. Interviews and coffee samples (n = 27) were collected from coffee shops in Singapore, Indonesia and India. In addition, 3000 men and women who were Chinese, Malay, and Indian residents of Singapore participated in a cross-sectional study. Results and Discussion The traditional 'sock' method of coffee preparation used in Singapore resulted in cafestol concentrations comparable to European paper drip filtered coffee (mean 0.09 ± SD 0.064 mg/cup). This amount would result in negligible predicted increases in serum cholesterol and triglyceride concentrations. Similarly low amounts of cafestol were found in Indian 'filter' coffee that used a metal mesh filter (0.05 ± 0.05 mg/cup). Coffee samples from Indonesia using the 'sock' method (0.85 ± 0.41 mg/cup) or a metal mesh filter (0.98 mg/cup) contained higher amounts of cafestol comparable to espresso coffee. Unfiltered coffee from Indonesia contained an amount of cafestol (4.43 mg/cup) similar to Scandinavian boiled, Turkish and French press coffee with substantial predicted increases in serum cholesterol (0.33 mmol/l) and triglycerides (0.20 mmol/l) concentrations for consumption of 5 cups per day. In the Singaporean population, higher coffee consumption was not substantially associated with serum lipid concentrations after adjustment for potential confounders [LDL-cholesterol: 3.07 (95% confidence interval 2.97-3.18) for coffee consumed in Singapore and India, coffee consumption in these countries does not appear

  18. Estudo retrospectivo de latrodectismo na Bahia, Brasil

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    Rejâne Maria Lira-da-Silva

    1995-09-01

    Full Text Available O trabalho apresenta um estudo retrospectivo de setenta e sete casos de latrodectismo no Estado da Bahia, Brasil, de agosto de 1980 a julho de 1990. Os dados foram levantados nos livros de registro e arquivo de fichas do CIAVE. O agente etiológico em 28% dos acidentes aracnídeos foi a espécie L. curacaviensis e a maior incidência foi registrada no meio urbano (57%, em indivíduos do sexo masculino (70% e faixa etária de 10 a 29 anos (58%. Os principais sinais locais foram dor (56%, pápula eritematosa (21% e edema discreto (17%, e os sistêmicos foram dor em membros inferiores (29%, tremores e contraturas (29%, sudorese (28% parestesia em membros (21% e dor abdominal (17%. O tratamento foi sintomático em 67% dos casos e específico em 21%. O tempo de permanência hospitalar após o uso do soro antilatrodectus foi menor que 24 horas em 64% dos casos.This work is a retrospective study of latrodectism in the State of Bahia, Brazil, from August 1980 to July 1990. The data concerning the accidents were obtained from file cards at the Antivenom Information Center of Bahia (AVICB. Latrodectus curacavienis was the ethiologic agent identified in 28% of the arachnid accidents. The major incidence was registered in urban area (57% affecting men (70% more than women, with 10 to 29year-old age group (58%. Local pain (56%, erythematous papula (29% and light oedema (17% were the principal local symptoms. Pain in the limbs (29%, tremor and rigidities (29%, sweating (28%, limbs and arms paresthesia (21% and abdominal pain (17% were systemic ones. The treatment was mainly symptomatic (67% and antivenin serum was used in 21% of the cases. After serotherapy, 64% of the patients left the hospital within less than 24 hours.

  19. Valor do estudo citogenetico no transexualismo

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    P. H. Saldanha

    1976-09-01

    Full Text Available O transexualismo é caracterizado como uma entidade psiquiátrica, distinta do homossexualismo e travestismo, revendo-se suas manifestações sindrômicas. Discutem-se as duas principais causas etiológicas plausíveis do transexualismo, a saber: a hipótese psicoanalítica fundamentada na regressão psicossexual com estampagem da figura materna e o modelo neuro-endócrino que pressupõe alterações nos centros de identidade sexual do hipotámo. Com base nesta última explicação propõe-se, a exemplo do que parece ocorrer na síndrome de Morris, cujas células (XY não respondem ao efeito masculinizante da testosterona plasmática, que os transexuais devem possuir mosaicismo detectável ou críptico, quanto aos cromossomos sexuais, nos centros hipotalâmicos de identidade sexual que não respondem à secreção androgênica produzida pela gônada primitiva. Esta possibilidade explicaria a excessiva prevalência da síndrome entre homens, bem como a sua manifestação com feições típicas no sexo masculino e ainda a ocorrência esporádica da síndrome. O estudo citogenético revelou que a frequência (32% de mosaicismo quanto aos cromossomos sexuais em 25 transexuais é estatisticamente superior aos valores observados em 14 homossexuais e 40 controles normais, nos quais a proporção de mosaicismo é praticamente nula. Considera-se a possibilidade do critério cariotípico constituir valioso subsídio na diagnose da síndrome.

  20. Histerectomias: estudo retrospectivo de 554 casos

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    Eddie Fernando Cândido Murta

    Full Text Available OBJETIVO: A histerectomia é uma operação muito realizada, entretanto há poucos trabalhos na literatura nacional sobre suas indicações, técnica e complicações. O objetivo deste trabalho é avaliar estes procedimentos realizados na Disciplina de Ginecologia e Obstetrícia da Faculdade de Medicina do Triângulo Mineiro. MÉTODO: Estudo retrospectivo de 470 histerectomias abdominais e 84 vaginais foi conduzido analisando as indicações, tempo de cirurgia e internação, tipo de incisão e morbidez. RESULTADOS: As principais indicações foram o mioma uterino e o prolapso uterino para as histerectomias abdominais e vaginais, respectivamente. As complicações intra-operatórias aconteceram em 3,4% e as pós-operatórias em 2,4% do total de casos. Nenhuma diferença estatística foi encontrada no número de complicações em relação ao tipo de incisão (vertical ou transversal. O tempo de cirurgia e o de hospitalização foram estatisticamente maiores nas incisões verticais. A hemorragia foi a mais freqüente complicação intra-operatória e a infecção da incisão operatória foi a mais freqüente no pós-operatório. CONCLUSÕES: A histerectomia é um procedimento de baixo risco, no entanto, a realização de revisões sobre indicações e complicações, e a pesquisa de melhores técnicas cirúrgicas são necessárias para torná-la cada vez mais segura.

  1. Reticulossarcomas epidurais intraespinais: estudo de 34 casos

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    Lígia M. B. Coutinho

    1974-12-01

    Full Text Available Foi realizado um estudo de 34 casos de reticulossarcomas localizados no tecido epidural intraespinal. Assinala-se sua maior freqüência entre homens com mais de 50 anos, embora pacientes mais jovens possam ser acometidos, principalmente entre 15 e 29 anos. A distribuição segmentar dos tumores permite um diagnóstico diferencial, sendo a região torácica a sede mais freqüentes de sarcomas. O principal sintoma encontrado nos pacientes com reticulossarcoma foi a dor, do tipo radicular, que serve para diferenciar os tumores intra e extra-medulares, tendo sido de aparecimento precoce em 15 dos pacientes estudados, precedendo em muitos meses e até anos os demais sintomas. Classificou-se os tumores reticulohistiocitários epidurais do canal vertebral em três grupos, de acordo com seu aspecto tecidual e celular e, principalmente, com a presença e distribuição da reticulina. Relacionou-se o tempo de evolução desses tumores com seu aspecto histopatológico, chegando-se à conclusão que os tumores classificados no grupo III apresentam um período evolutivo maior que os demais, o que nos levou a pensar que crescem mais lentamente, apresentando, portanto, melhor prognóstico que os demais tipos, muito embora todos os tipos de reticulossarcomas possam apresentar um prognóstico mais favorável quando precocemente diagnosticados e tratados adequadamente.

  2. Estudos sobre a Esquistosomose em Pernambuco, Brasil

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    Aggeu Magalhães

    1940-01-01

    Full Text Available Os autores referem os resultados dos estudos sobre esquistosomose realisados, em Pernambuco, de Novembro de 1938 a Dezembro de 1939. Não se extendem em discussão, nem comentarios, tampouco, relacionam suas verificações com o que já foi relatado sobre o assunto em trabalhos nacionais e estrangeiros, pelo fato de que, não se trata de um resultado definitivo e sim de atividades que irão proseguir. Os estudos sistematicos em torno da endemia esquistosomica foram executados em duas localidades diversas pela população e pela situação geografica e ecologica. Pontezinha é uma povoação de 1200 habitantes localizada proximo a um conjunto de charcos e lagõas de agua dôce onde pululam caramujos do tipo olivaceus de Spix, e apresenta uma taxa de infestação de 21,4% para individuos do sexo masculino, e de 14,7% para os do sexo feminino. A distribuição por grupos de idade mostra que a incidencia cresce até o grupo de 16 a 20 anos de idade, quando atinge a percentagem de 37,8%, para decrescer em seguida. Em Vitoria a incidencia foi maior nos indivíduos do sexo feminino, atingindo a taxa de 41,4%, dando o sexo masculino a taxa de 36,9%. Por idade a percentagem maxima é atingida pelo grupo de 11 a 15 anos. Vitoria é uma cidade de 1500 habitantes, construida á margem do rio Tapacurá, onde se encontram numerosos caramujos do tipo centimetralis Lutz. A divergencia de incidencia corre por conta dos habitos da população e pela utilização que ela faz do rio, sendo maior em Vitoria porque a proximidade do rio facilita o uso de suas aguas para banho e serviços domesticos, condicionando este ultimo fato, a maior infestação das mulheres. O estudo da frequencia da infestação dos caramujos por cercarias de diversos trematodios, permite suspeitar uma relação inversa entre a taxa de infestação e o diametro maximo atingido pelos caramujos, parecendo este fato confirmar os trabalhos de Vianna Martins sobre a identidade dos hospedeiros

  3. Estudos organizacionais, (descolonialidade e estudos da dependência: as contribuições da Cepal

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    Sergio Wanderley

    Full Text Available O objetivo deste artigo é (revisitar o conceito de dependência como categoria de investigação da (e a partir da América Latina, por meio de uma perspectiva histórica crítica descolonial, a fim de ampliar o espaço de debates em estudos organizacionais e promover alternativas à ordem neoliberal. Este ensaio terá como foco os estudos da dependência realizados pela Comissão Econômica para a América Latina e o Caribe (Cepal durante a década de 1950 e o início dos anos 1960. A perspectiva descolonial é um constructo teórico de autores latino-americanos que se consideram herdeiros da longa tradição do pensamento social crítico da região, na qual a teoria da dependência está inserida. As propostas dos autores da dependência confrontaram as principais teorias ortodoxas do Norte produzidas à época. A partir do conceito de centro-periferia, da denúncia de assimetrias nas relações entre essas regiões, do reconhecimento da interdependência entre desenvolvimento e subdesenvolvimento, o conceito de dependência foi sendo (reelaborado nas décadas de 1950 e 1960 por diversos autores latino-americanos como uma categoria de investigação da realidade da (e a partir da região. Essas investigações tinham por objetivo não somente elaborar constructos teóricos, mas, também, transformar a realidade por meio da criação de diversas organizações e instituições que deveriam servir ao propósito de superar o subdesenvolvimento. A denúncia da historicidade da situação de subdesenvolvimento desnudou o caráter de neutralidade do economicismo das teorias produzidas no Norte e promoveu um encontro teórico entre economia e política que muito tem a contribuir com a área dos estudos organizacionais.

  4. Microbial transformations of isocupressic acid.

    Science.gov (United States)

    Lin, S J; Rosazza, J P

    1998-07-01

    Microbial transformations of the labdane-diterpene isocupressic acid (1) with different microorganisms yielded several oxygenated metabolites that were isolated and characterized by MS and NMR spectroscopic analyses. Nocardia aurantia (ATCC 12674) catalyzed the cleavage of the 13,14-double bond to yield a new nor-labdane metabolite, 2. Cunninghamella elegans (-) (NRRL 1393) gave 7beta-hydroxyisocupressic acid (3) and labda-7,13(E)-diene-6beta,15, 17-triol-19-oic acid (4), and Mucor mucedo (ATCC 20094) gave 2alpha-hydroxyisocupressic acid (5) and labda-8(17),14-diene-2alpha, 13-diol-19-oic acid (6).

  5. ESTUDOS NUTRICIONAIS COM ARROZ (Oryza sativa, L.

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    Alfredo LAM-SÁNCHEZ

    2009-07-01

    Full Text Available

    RESUMO: Estudos com arroz (Oryza sativa, L. foram realizados a fim de determinar a variabilidade do teor de proteína em coleções de cultivares e a possibilidade de utilizar as camadas periféricas do grão na alimentação humana. Foram analisadas 626 linhagens e cultivares de arroz que mostraram uma variação quanto ao teor de proteína de 4.00 a 14.31 g/100 g de amostra. Para se extrair as camadas periféricas do grão, foi desenvolvida uma máquina experimental, que removia as camadas superficiais dos grãos de arroz integral (inteiros somente com a casca removida. Verificou-se que a quantidade de proteína do grão diminui de fora para dentro. O resíduo, que era constituído de grande parte de restos de endosperma com um teor protéico menor mas incluía o embrião, que é a primeira estrutura a se deslocar de grão no beneficiamento e contém proteínas, vitaminas, sais minerais e lípides, tinha um bom valor nutritivo. Desenvolveu-se também uma fórmula para alimentação infantil a partir da mistura de leite de vaca com arroz, com composição semelhante à do leite materno, a ser utilizada sob a forma de bebida ou mingau. Formulações com 40%-20% arroz + 60%-80% leite de vaca proporcionaram valores de Coeficiente de Eficiência Protéica (2,39 e 2,38, respectivamente que não diferiam estatisticamente do valor de 2,43 obtido na fórmula com 100% de leite de vaca. Seus preços foram 59% e 65% do obtido para o leite de vaca integral. PALAVRAS-CHAVE: Arroz; qualidade do grão; proteína; camadas periféricas; misturas arroz-leite; nutrição humana.

  6. Estudo de caso: disfonia psicogênica

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    Marcela Bergamini

    2015-02-01

    Full Text Available Disfonia funcional psicogênica possui relação com aspectos psicológicos. O tipo de voz, a articulação e a fluência são sensíveis às oscilações psicológicas. Uma voz em ajuste de falsete paralítico, caracterizada como uma voz de pitch extremamente agudo pode ser encontrada em casos de disfonias psicogênicas. Em qualquer quadro psicogênico é essencial um diagnóstico diferencial, a fim de viabilizar um tratamento fonoaudiológico e médico adequados e efetivos. O prognóstico de uma alteração psicogênica geralmente é bom. O quadro de disfonia psicogênica é mais comum no sexo feminino. O objetivo desse estudo é apresentar a evolução terapêutica de um indivíduo com disfonia psicogênica. A.J.O., 45 anos, sexo masculino, compareceu ao Ambulatório de Avaliação e Terapia de Voz encaminhado por médico otorrinolaringologista. Trouxe a queixa vocal "pela manhã a voz está perfeita, conforme o dia passa, a voz fica fina". Realizou avaliação fonoaudiológica em 18 de abril de 2013, com laudo médico de "mobilidade reduzida de prega vocal esquerda". Tratamento com técnica de sons disparadores, com modificação direta nos sintomas e evolução rápida. Demonstrou-se satisfeito com sua qualidade vocal. Em casos de suspeita de disfonias psicogênicas de conversão, devem-se descartar alterações orgânicas, sendo essencial uma avaliação fonoaudiológica que auxilie no diagnóstico diferencial e provas terapêuticas que incentivem o paciente.

  7. HpDTC1, a Stress-Inducible Bifunctional Diterpene Cyclase Involved in Momilactone Biosynthesis, Functions in Chemical Defence in the Moss Hypnum plumaeforme

    Science.gov (United States)

    Okada, Kazunori; Kawaide, Hiroshi; Miyamoto, Koji; Miyazaki, Sho; Kainuma, Ryosuke; Kimura, Honoka; Fujiwara, Kaoru; Natsume, Masahiro; Nojiri, Hideaki; Nakajima, Masatoshi; Yamane, Hisakazu; Hatano, Yuki; Nozaki, Hiroshi; Hayashi, Ken-ichiro

    2016-01-01

    Momilactones, which are diterpenoid phytoalexins with antimicrobial and allelopathic functions, have been found only in rice and the moss Hypnum plumaeforme. Although these two evolutionarily distinct plant species are thought to produce momilactones as a chemical defence, the momilactone biosynthetic pathway in H. plumaeforme has been unclear. Here, we identified a gene encoding syn-pimara-7,15-diene synthase (HpDTC1) responsible for the first step of momilactone biosynthesis in the moss. HpDTC1 is a bifunctional diterpene cyclase that catalyses a two-step cyclization reaction of geranylgeranyl diphosphate to syn-pimara-7,15-diene. HpDTC1 transcription was up-regulated in response to abiotic and biotic stress treatments. HpDTC1 promoter-GUS analysis in transgenic Physcomitrella patens showed similar transcriptional responses as H. plumaeforme to the stresses, suggesting that a common response system to stress exists in mosses. Jasmonic acid (JA), a potent signalling molecule for inducing plant defences, could not activate HpDTC1 expression. In contrast, 12-oxo-phytodienoic acid, an oxylipin precursor of JA in vascular plants, enhanced HpDTC1 expression and momilactone accumulation, implying that as-yet-unknown oxylipins could regulate momilactone biosynthesis in H. plumaeforme. These results demonstrate the existence of an evolutionarily conserved chemical defence system utilizing momilactones and suggest the molecular basis of the regulation for inductive production of momilactones in H. plumaeforme. PMID:27137939

  8. Antioxidant and antimicrobial effects of dietary supplementation with rosemary diterpenes (carnosic acid and carnosol) vs vitamin E on lamb meat packed under protective atmosphere.

    Science.gov (United States)

    Ortuño, Jordi; Serrano, Rafael; Bañón, Sancho

    2015-12-01

    The antioxidant and antimicrobial effects on lamb meat of the dietary use of rosemary diterpenes and vitamin E were compared. Thirty fattening lambs were assigned to three diets: (C) control; (R) C plus 600 mg kg(-1) carnosic acid and carnosol at 1:1 w:w; or (E) C plus 600 mg kg(-1) α-tocopherol. The deposition of the dietary supplements in the muscle was determined. Microbial quality (total viable counts, Lactic Acid Bacteria, Enterobacteriaceae, Escherichia coli and Salmonella spp), oxidative stability (CIELab color, malondialdehyde and total carbonyls) and sensory attributes (appearance and odor) were determined in loin stored at 2°C under 70% O2/30% CO2 atmosphere. Microbial quality was ensured by packaging and chilling. The E-diet was more effective (P ≤ 0.05) than the R-diet in preventing meat oxidation, although the latter had antimicrobial effects on meat. The shelf life of lamb (assessed as the loss of freshness) could be increased by 5 (R-diet) or 10 (E-diet) days.

  9. Abietane diterpenes from the cones of Abies numidica de Lannoy ex Carrière (Pinaceae) and in vitro evaluation of their antimicrobial properties.

    Science.gov (United States)

    Belhadj Mostefa, Maya; Abedini, Amin; Voutquenne-Nazabadioko, Laurence; Gangloff, Sophie C; Kabouche, Ahmed; Kabouche, Zahia

    2017-03-01

    Eight known abietane diterpenes (1-8) were isolated for the first time from Abies numidica cones (Pinaceae). The structures of all compounds were established by means of 1D and 2D NMR, and UV spectral analyses. The hydromethanolic extract of A. numidica cones was tested for its antimicrobial activity against 17 gram-positive and gram-negative bacteria and against five yeasts by the use of liquid and solid medium and bioautography methods. The best antimicrobial activity was found against gram-positive bacteria (MIC ≤ 0.3 mg/mL) Bacillus subtilis, Enterococcus faecalis ATCC 1034, Staphylococcus aureus 8325.4, S. aureus CIP 53.154, Micrococcus luteus and Listeria innocua and against Candida yeasts. The determination of MIC's of isolated products showed a high activity of compounds 4 and 6 against S. aureus, L. innocua (MIC = 62.5 μg/mL) and E. faecalis (MIC = 125 μg/mL).

  10. A pseudopterane diterpene isolated from the octocoral Pseudopterogorgia acerosa inhibits the inflammatory response mediated by TLR-ligands and TNF-alpha in macrophages.

    Science.gov (United States)

    González, Yisett; Doens, Deborah; Santamaría, Ricardo; Ramos, Marla; Restrepo, Carlos M; Barros de Arruda, Luciana; Lleonart, Ricardo; Gutiérrez, Marcelino; Fernández, Patricia L

    2013-01-01

    Several diterpenoids isolated from terrestrial and marine environments have been identified as important anti-inflammatory agents. Although considerable progress has been made in the area of anti-inflammatory treatment, the search for more effective and safer compounds is a very active field of research. In this study we investigated the anti-inflammatory effects of a known pseudopterane diterpene (referred here as compound 1) isolated from the octocoral Pseudopterogorgia acerosa on the tumor necrosis factor- alpha (TNF-α) and TLRs- induced response in macrophages. Compound 1 inhibited the expression and secretion of the inflammatory mediators TNF-α, interleukin (IL)-6, IL-1β, nitric oxide (NO), interferon gamma-induced protein 10 (IP-10), ciclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) and monocyte chemoattractant protein-1 (MCP-1) induced by LPS in primary murine macrophages. This effect was associated with the inhibition of IκBα degradation and subsequent activation of NFκB. Compound 1 also inhibited the expression of the co-stimulatory molecules CD80 and CD86, which is a hallmark of macrophage activation and consequent initiation of an adaptive immune response. The anti-inflammatory effect was not exclusive to LPS because compound 1 also inhibited the response of macrophages to TNF-α and TLR2 and TLR3 ligands. Taken together, these results indicate that compound 1 is an anti-inflammatory molecule, which modulates a variety of processes occurring in macrophage activation.

  11. Abietane diterpene acids from the bark of lllicium jiadifengpi%假地枫皮中二萜酸类化合物研究

    Institute of Scientific and Technical Information of China (English)

    黄平; Gloria Karagianis; Peter G Waterman

    2005-01-01

    Five diterpene acids were isolated from the bark of Illicium jiadifengpi (Illiciaceae). On the basis of spectral data(1H NMR and 13C NMR, 1H-1H COSY and NOESY, HSQC and HMBC), their structures were elucidated as 4-epi-dehydroabietic acid (1), 4-epi-sandaracopinaric acid (2), 4- epi-abietic acid ( 3 ), 4- epi-isopimaric acid ( 4 ) and 8,11,13, 15-abietatetraen19-oic acid (5).%从八角属植物假地枫皮的石油醚提取物中分离出5个二萜酸类化合物,经波谱数据分析(1H NMR、13C NMR、1H-1H COSY、NOESY、HSQC和HMBC),分别鉴定为4-epi-dehydroabietic acid(1)、4-epi-sandaracopinaric acid (2)、4-epi-abietic acid(3)、4-epi-isopimaric acid(4)和8,11,13,15-abietatetraen-19-oic acid(4).

  12. A pseudopterane diterpene isolated from the octocoral Pseudopterogorgia acerosa inhibits the inflammatory response mediated by TLR-ligands and TNF-alpha in macrophages.

    Directory of Open Access Journals (Sweden)

    Yisett González

    Full Text Available Several diterpenoids isolated from terrestrial and marine environments have been identified as important anti-inflammatory agents. Although considerable progress has been made in the area of anti-inflammatory treatment, the search for more effective and safer compounds is a very active field of research. In this study we investigated the anti-inflammatory effects of a known pseudopterane diterpene (referred here as compound 1 isolated from the octocoral Pseudopterogorgia acerosa on the tumor necrosis factor- alpha (TNF-α and TLRs- induced response in macrophages. Compound 1 inhibited the expression and secretion of the inflammatory mediators TNF-α, interleukin (IL-6, IL-1β, nitric oxide (NO, interferon gamma-induced protein 10 (IP-10, ciclooxygenase (COX-2, inducible nitric oxide synthase (iNOS and monocyte chemoattractant protein-1 (MCP-1 induced by LPS in primary murine macrophages. This effect was associated with the inhibition of IκBα degradation and subsequent activation of NFκB. Compound 1 also inhibited the expression of the co-stimulatory molecules CD80 and CD86, which is a hallmark of macrophage activation and consequent initiation of an adaptive immune response. The anti-inflammatory effect was not exclusive to LPS because compound 1 also inhibited the response of macrophages to TNF-α and TLR2 and TLR3 ligands. Taken together, these results indicate that compound 1 is an anti-inflammatory molecule, which modulates a variety of processes occurring in macrophage activation.

  13. HpDTC1, a Stress-Inducible Bifunctional Diterpene Cyclase Involved in Momilactone Biosynthesis, Functions in Chemical Defence in the Moss Hypnum plumaeforme.

    Science.gov (United States)

    Okada, Kazunori; Kawaide, Hiroshi; Miyamoto, Koji; Miyazaki, Sho; Kainuma, Ryosuke; Kimura, Honoka; Fujiwara, Kaoru; Natsume, Masahiro; Nojiri, Hideaki; Nakajima, Masatoshi; Yamane, Hisakazu; Hatano, Yuki; Nozaki, Hiroshi; Hayashi, Ken-Ichiro

    2016-05-03

    Momilactones, which are diterpenoid phytoalexins with antimicrobial and allelopathic functions, have been found only in rice and the moss Hypnum plumaeforme. Although these two evolutionarily distinct plant species are thought to produce momilactones as a chemical defence, the momilactone biosynthetic pathway in H. plumaeforme has been unclear. Here, we identified a gene encoding syn-pimara-7,15-diene synthase (HpDTC1) responsible for the first step of momilactone biosynthesis in the moss. HpDTC1 is a bifunctional diterpene cyclase that catalyses a two-step cyclization reaction of geranylgeranyl diphosphate to syn-pimara-7,15-diene. HpDTC1 transcription was up-regulated in response to abiotic and biotic stress treatments. HpDTC1 promoter-GUS analysis in transgenic Physcomitrella patens showed similar transcriptional responses as H. plumaeforme to the stresses, suggesting that a common response system to stress exists in mosses. Jasmonic acid (JA), a potent signalling molecule for inducing plant defences, could not activate HpDTC1 expression. In contrast, 12-oxo-phytodienoic acid, an oxylipin precursor of JA in vascular plants, enhanced HpDTC1 expression and momilactone accumulation, implying that as-yet-unknown oxylipins could regulate momilactone biosynthesis in H. plumaeforme. These results demonstrate the existence of an evolutionarily conserved chemical defence system utilizing momilactones and suggest the molecular basis of the regulation for inductive production of momilactones in H. plumaeforme.

  14. A Pseudopterane Diterpene Isolated From the Octocoral Pseudopterogorgia acerosa Inhibits the Inflammatory Response Mediated by TLR-Ligands and TNF-Alpha in Macrophages

    Science.gov (United States)

    González, Yisett; Doens, Deborah; Santamaría, Ricardo; Ramos, Marla; Restrepo, Carlos M.; Barros de Arruda, Luciana; Lleonart, Ricardo; Gutiérrez, Marcelino; Fernández, Patricia L.

    2013-01-01

    Several diterpenoids isolated from terrestrial and marine environments have been identified as important anti-inflammatory agents. Although considerable progress has been made in the area of anti-inflammatory treatment, the search for more effective and safer compounds is a very active field of research. In this study we investigated the anti-inflammatory effects of a known pseudopterane diterpene (referred here as compound 1) isolated from the octocoral Pseudopterogorgia acerosa on the tumor necrosis factor- alpha (TNF-α) and TLRs- induced response in macrophages. Compound 1 inhibited the expression and secretion of the inflammatory mediators TNF-α, interleukin (IL)-6, IL-1β, nitric oxide (NO), interferon gamma-induced protein 10 (IP-10), ciclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) and monocyte chemoattractant protein-1 (MCP-1) induced by LPS in primary murine macrophages. This effect was associated with the inhibition of IκBα degradation and subsequent activation of NFκB. Compound 1 also inhibited the expression of the co-stimulatory molecules CD80 and CD86, which is a hallmark of macrophage activation and consequent initiation of an adaptive immune response. The anti-inflammatory effect was not exclusive to LPS because compound 1 also inhibited the response of macrophages to TNF-α and TLR2 and TLR3 ligands. Taken together, these results indicate that compound 1 is an anti-inflammatory molecule, which modulates a variety of processes occurring in macrophage activation. PMID:24358331

  15. Part 1. Approaches to the total synthesis of the diterpene marrubin. Part II. Ligand assisted nucleophilic additions. Part III. Coal processing in a non-dissolving medium

    Energy Technology Data Exchange (ETDEWEB)

    Lipscombe, J.; Charles, W. II

    1987-01-01

    The total synthesis of the diterpene marrubiin was attempted using methodology previously developed in the Liotta laboratories. Ligand Assisted Nucleophilic Additions (LANA) comprise a new class of synthetic reactions involving the stereoselective addition of some group, usually in an intramolecular conjugate fashion. This group is itself derived from an earlier nucleophilic addition to a carbonyl moiety. Exchange of the original metal of the alkoxide with a second metal (which must be at least divalent) having a transferrable group demands that the stereochemistry of the first addition control the second. The general features of these types of processes are discussed. Initial work with substituted quinones is presented, and some mechanistic rationales are given. A novel method of coal processing at modest temperatures and hydrogen pressures (250-300/sup 0/C, 8.0 MPa) is discussed. Coals reacted with small amounts of tetrahydroquinoline in the presence of fluorocarbons or other non-dissolving media underwent astounding increases in pyridine solubility, a common standard in coal liquefaction studies. Several additives were employed, although only tetrahydroquinoline was found to effectively increase coal solubility. Coals having vastly different individual properties behaved similarly under these conditions. Kinetic and model studies were used in an attempt to clarify the reactions occurring under these novel conditions.

  16. The clerodane diterpene casearin J induces apoptosis of T-ALL cells through SERCA inhibition, oxidative stress, and interference with Notch1 signaling.

    Science.gov (United States)

    De Ford, C; Heidersdorf, B; Haun, F; Murillo, R; Friedrich, T; Borner, C; Merfort, I

    2016-01-28

    T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematologic malignancy that preferentially affects children and adolescents. Over 50% of human T-ALLs possess activating mutations of Notch1. The clerodane diterpene casearin J (CJ) is a natural product that inhibits the sarcoendoplasmatic reticulum calcium ATPase (SERCA) pump and induces cell death in leukemia cells, but the molecular mechanism of cytotoxicity remains poorly understood. Here we show that owing to SERCA pump inhibition, CJ induces depletion of the endoplasmic reticulum calcium pools, oxidative stress, and apoptosis via the intrinsic signaling pathway. Moreover, Notch1 signaling is reduced in T-ALL cells with auto-activating mutations in the HD-domain of Notch1, but not in cells that do not depend on Notch1 signaling. CJ also provoked a slight activation of NF-κB, and consistent with this notion a combined treatment of CJ and the NF-κB inhibitor parthenolide (Pt) led to a remarkable synergistic cell death in T-ALL cells. Altogether, our data support the concept that inhibition of the SERCA pump may be a novel strategy for the treatment of T-ALL with HD-domain-mutant Notch1 receptors and that additional treatment with the NF-κB inhibitor parthenolide may have further therapeutic benefits.

  17. Redox-dependent induction of antioxidant defenses by phenolic diterpenes confers stress tolerance in normal human skin fibroblasts: Insights on replicative senescence.

    Science.gov (United States)

    Carvalho, Ana C; Gomes, Andreia C; Pereira-Wilson, Cristina; Lima, Cristovao F

    2015-06-01

    Mild stress-induced hormesis represents a promising strategy for targeting the age-related accumulation of molecular damage and, therefore, for preventing diseases and achieving healthy aging. Fruits, vegetables, and spices contain a wide variety of hormetic phytochemicals, which may explain the beneficial health effects associated with the consumption of these dietary components. In the present study, the induction of cellular antioxidant defenses by the phenolic diterpenes carnosic acid (CA) and carnosol (CS) were studied in normal human skin fibroblasts, and insights into the aging process at the cellular level investigated. We observed that CA and CS induced several cytoprotective enzymes and antioxidant defenses in human fibroblasts, whose induction was dependent on the cellular redox state for CS and associated with Nrf2 signaling for both compounds. The stress response elicited by preincubation with CS conferred a cytoprotective action against a following oxidant challenge with tert-butyl hydroperoxide, confirming its hormetic effect. Preincubation of normal fibroblasts with CS also protected against hydrogen peroxide-induced premature senescence. Furthermore, cultivation of middle passage normal human skin fibroblasts in the presence of CS ameliorated the physiological state of cells during replicative senescence. Our results support the view that mild stress-induced antioxidant defenses by CS can confer stress tolerance in normal cells and may have important implications in the promotion of healthy aging.

  18. Uma ferramenta para planejamento de estudos para concursos

    Directory of Open Access Journals (Sweden)

    Leonardo Pimentel

    2015-11-01

    Full Text Available De acordo com a constituição brasileira, a única forma de se tornar um empregado do governo é passar em um concurso público. O exame de conhecimento do cargo é um dos componentes mais importantes de um concurso. Estudar para esse exame é uma tarefa que requer planejamento. Um meio de obter um bom planejamento é construir uma grade de horários que relacione as matérias a serem estudadas com os respectivos horários disponíveis para o estudo. Esse tipo de problema se assemelha bastante à gerência de projetos, em que se procura estabelecer a distribuição das atividades ao longo do tempo disponível. Assim, torna-se interessante explorar a dinâmica de estudos para concursos como um projeto. O presente trabalho apresenta a ferramenta de apoio ao estudos - Passe-me, baseada no ciclo PDCA (do inglês, Plan-Do-Check-Act. Por se tratar de uma ferramenta web, o tempo de resposta para produzir uma grade de estudos é fundamental. A ferramenta faz uso de algoritmo genético para calcular a grade de horários priorizando a velocidade de resposta. A ferramenta foi avaliada em diferentes cenários de uso e obteve desempenho satisfatório durante a produção da grade de estudos.

  19. Desenvolvimento da Tanatologia: estudos sobre a morte e o morrer

    Directory of Open Access Journals (Sweden)

    Maria Julia Kovács

    2008-12-01

    Full Text Available Este artigo discute os principais temas e pesquisas na área da Tanatologia, estudos sobre a morte e o morrer. São apresentados os autores pioneiros que escreveram as primeiras obras de sistematização da Tanatologia entre os quais: Herman Feifel, Robert Kastenbaum e Elizabeth Kübler-Ross, e os principais temas de estudo: luto, violência e guerra, a morte e a TV, cuidados a pacientes gravemente enfermos, além da formação de profissionais da área de saúde e educação para lidar com pessoas vivendo situações de perdas e morte. São feitas propostas de estudos para o futuro desenvolvimento da Tanatologia em nosso país.

  20. BASES DO COMPROMETIMENTO ORGANIZACIONAL: UM ESTUDO COMPARATIVO EM EMPRESAS JUNIORES

    Directory of Open Access Journals (Sweden)

    Maria José da Silva Feitosa

    2013-04-01

    Full Text Available Nos últimos anos intensificaram-se os estudos referentes às bases do comprometimento organizacional, contempladas no modelo Meyer e Allen. Nessa perfectiva, o presente estudo objetiva verificar as bases afetiva, normativa e instrumental em duas empresas de consultoria. Para tanto, foi realizada uma pesquisa descritiva, de abordagem quantitativa. Os resultados apontam que a base afetiva se destacou em ambas as empresas, tanto em uma análise por gênero, quanto em uma perspectiva geral. Esse resultado pode estar relacionado à possibilidade de que os estudantes que ingressam em empresas juniores se entusiasmem com a oportunidade de aplicar seus conhecimentos e se comprometam de forma afetiva com a organização. No entanto, para testar essa relação sugere-se a realização de novos estudos.

  1. Desenvolvimento da Tanatologia: estudos sobre a morte e o morrer

    OpenAIRE

    Maria Julia Kovács

    2008-01-01

    Este artigo discute os principais temas e pesquisas na área da Tanatologia, estudos sobre a morte e o morrer. São apresentados os autores pioneiros que escreveram as primeiras obras de sistematização da Tanatologia entre os quais: Herman Feifel, Robert Kastenbaum e Elizabeth Kübler-Ross, e os principais temas de estudo: luto, violência e guerra, a morte e a TV, cuidados a pacientes gravemente enfermos, além da formação de profissionais da área de saúde e educação para lidar com pessoas vivend...

  2. Estudo patográfico de Fernando Pessoa

    OpenAIRE

    2009-01-01

    Fernando Pessoa, incontestavelmente um dos maiores gênios da literatura universal, é objeto deste estudo patográfico. Através da análise de sua biografia e obra, os autores buscam delinear seus perfis psicológico e psicopatológico e caracterizar uma associação entre sua evidente bipolaridade e seu padrão criativo. Os dados do estudo revelam claramente um componente bipolar e sugerem haver influência de seu humor de base sobre a atividade literária, quanto ao conteúdo, número de po...

  3. Desenvolvimento da Tanatologia: estudos sobre a morte e o morrer

    OpenAIRE

    Maria Julia Kovács

    2008-01-01

    Este artigo discute os principais temas e pesquisas na área da Tanatologia, estudos sobre a morte e o morrer. São apresentados os autores pioneiros que escreveram as primeiras obras de sistematização da Tanatologia entre os quais: Herman Feifel, Robert Kastenbaum e Elizabeth Kübler-Ross, e os principais temas de estudo: luto, violência e guerra, a morte e a TV, cuidados a pacientes gravemente enfermos, além da formação de profissionais da área de saúde e educação para lidar com pessoas vivend...

  4. 比色法测定昆明山海棠片中总二萜内酯的含量%Determination the Content of Total Diterpene Lactones in Kunming Shan-haitang Tablets by Colorimetry

    Institute of Scientific and Technical Information of China (English)

    江银蒜; 刘岩庭; 吴春敏

    2015-01-01

    目的 建立测定昆明山海棠片中总二萜内酯含量的方法. 方法 采用比色法,在545nm波长处进行总二萜内酯的含量测定. 结果比色法测定昆明山海棠片总二萜内酯(以雷公藤甲素计)在8.17μg· mL-1 ~49.1μg· mL -1范围内线性关系良好(r=0.9999),平均加样回收率为99.2%,RSD为1.94%(n=6). 结论 本法简便、准确,适用于昆明山海棠片中总二萜内酯的含量测定,不同企业的昆明山海棠片中总二萜内酯的含量差异较大,在临床用药时应引起重视.%OBJECTIVE To determine the content of total diterpene lactones in Kunming Shanhaitang tab-lets.METHODS A colorimetric method was performed to determine the content of total diterpene lactones at 545nm.RESULTS The calibration curves were linear in the range from 8.17μg· mL-1 to 49.1μg· mL-1 (r=0.9999 ) for total diterpene lactones ( calculated as triptolide ) with colorimetric method.The average recovery rate (n=6) was 99.2%with RSD as 1.94%.CONCLUSION The method is simple and accurate,and can be used in the determination of Kunming Shanhaitang tablets.The content of total diterpene lactones in Kunming Shanhaitang tablets from different factories were compraratively largely disparity,which should be taken into account in clinic.

  5. HPLC法测定千金子不同极性部位中二萜酯类化合物含量%Comparison of Content of Diterpene Esters from Different Polar Fractions in Euphorbiae Semen by HPLC Method

    Institute of Scientific and Technical Information of China (English)

    张超; 朱娟娟; 王英姿; 孙秀梅

    2014-01-01

    目的:考察千金子素L8、千金子素L1、千金子素L2、千金子素L3四种化合物在千金子不同极性部位的分布情况。方法:用HPLC测定,色谱柱为Inertsil ODS-3(250 mm×4.6 mm,5μm),流动相为甲醇-水(75∶25),流速1.0 mL·min-1,柱温25℃;检测波长275 nm。结果:4个二萜酯类化合物在石油醚部位中含量最高,在乙酸乙酯和正丁醇部位中较少,在剩余水部位基本检测不到二萜酯类化合物。结论:该方法准确简便,重现性好,可用于比较千金子不同极性部位二萜酯类化合物的分布情况。%Objective:To study the distribution of euphorbia factor L8,euphorbia factor L1,euphorbia factor L2 and euphorbia factor L3 in the different polar fractions of Euphorbiae semen. Methods:Using HPLC method determine the content of 4 ingredients. The column is performed on Inertsil ODS-3(250 mm×4.6 mm,5μm), with mobile phase for methanol and water(75∶25),velocity 1.0 mL·min-1,column temperature 25℃,275 nm wavelength detection. Results:The content of diterpene esters was the highest in the petroleum ether fraction of Euphorbiae semen. The content of diterpene esters was lower in the fractions of acetic ether and n-bu-tanol. In the remaining part of water,the diterpene esters were undetected. Conclusion:The method is sim-ple,accurate and well reproducibile,so it can be used to compare the content of diterpene esters from dif-ferent polar fractions in Euphorbiae semen.

  6. Kaurene diterpene induces apoptosis in U87 human malignant glioblastoma cells by suppression of anti-apoptotic signals and activation of cysteine proteases

    Energy Technology Data Exchange (ETDEWEB)

    Lizarte, F.S. Neto; Tirapelli, D.P.C. [Universidade de São Paulo, Departamento de Cirurgia e Anatomia, Faculdade de Medicina de Ribeirão Preto, Ribeirão Preto, SP (Brazil); Ambrosio, S.R. [Universidade de Franca, Núcleo de Pesquisa em Ciências e Tecnologia, Franca, SP (Brazil); Tirapelli, C.R. [Universidade de São Paulo, Laboratório de Farmacologia, Departamento de Enfermagem Psiquiátrica e Ciências Humanas, Escola de Enfermagem de Ribeirão Preto, Ribeirão Preto, SP (Brazil); Oliveira, F.M. [Universidade de São Paulo, Departamento de Clínica Médica, Faculdade de Medicina de Ribeirão Preto, Ribeirão Preto, SP (Brazil); Novais, P.C. [Universidade de São Paulo, Departamento de Cirurgia e Anatomia, Faculdade de Medicina de Ribeirão Preto, Ribeirão Preto, SP (Brazil); Peria, F.M.; Oliveira, H.F. [Universidade de São Paulo, Departamento de Clínica Médica, Faculdade de Medicina de Ribeirão Preto, Ribeirão Preto, SP (Brazil); Carlotti, C.G. Junior; Tirapelli, L.F. [Universidade de São Paulo, Departamento de Cirurgia e Anatomia, Faculdade de Medicina de Ribeirão Preto, Ribeirão Preto, SP (Brazil)

    2013-01-11

    Gliomas are the most common and malignant primary brain tumors in humans. Studies have shown that classes of kaurene diterpene have anti-tumor activity related to their ability to induce apoptosis. We investigated the response of the human glioblastoma cell line U87 to treatment with ent-kaur-16-en-19-oic acid (kaurenoic acid, KA). We analyzed cell survival and the induction of apoptosis using flow cytometry and annexin V staining. Additionally, the expression of anti-apoptotic (c-FLIP and miR-21) and apoptotic (Fas, caspase-3 and caspase-8) genes was analyzed by relative quantification (real-time PCR) of mRNA levels in U87 cells that were either untreated or treated with KA (30, 50, or 70 µM) for 24, 48, and 72 h. U87 cells treated with KA demonstrated reduced viability, and an increase in annexin V- and annexin V/PI-positive cells was observed. The percentage of apoptotic cells was 9% for control cells, 26% for cells submitted to 48 h of treatment with 50 µM KA, and 31% for cells submitted to 48 h of treatment with 70 µM KA. Similarly, in U87 cells treated with KA for 48 h, we observed an increase in the expression of apoptotic genes (caspase-8, -3) and a decrease in the expression of anti-apoptotic genes (miR-21 and c-FLIP). KA possesses several interesting properties and induces apoptosis through a unique mechanism. Further experiments will be necessary to determine if KA may be used as a lead compound for the development of new chemotherapeutic drugs for the treatment of primary brain tumors.

  7. Lobolide, a diterpene, blockades the NF-κB pathway and p38 and ERK MAPK activity in macrophages in vitro

    Institute of Scientific and Technical Information of China (English)

    Xiao-fen LV; Si-han CHEN; Jie LI; Jian-ping FANG; Yue-wei GUO; Kan DING

    2012-01-01

    Aim:Recent studies have shown that constitutive activation of the nuclear factor κB (NF-κB) plays a key role in chronic inflammation and cancers.The aim of this study was to characterize lobolide,a cembrane diterpene,as a drug candidate targeting the NF-κBsignaling pathway.Methods:A HEK 293/NF-KB-Luc stable cell line was constructed to evaluate the effect of lobolide on NF-κB activation.THP-1 human monocytes and peripheral blood mononuclear cells (PBMCs) from healthy volunteers were tested.Lipopolysaccharide (LPS)-induced TNFα and IL-1β production and activation of the TAK1-IKK-NF-κB pathway were studied using ELISA and Western blot analysis.Results:In HEK 293/NF-kB-Luc stable cells,Iobolide (0.19-50 μmol/L) inhibited NF-kB activation in a concentration-dependent manner with an IC50 value of 4.2±0.3 μmol/L.Treatment with Iobolide (2.5-10 μmol/L) significantly suppressed LPS-induced production of TNFα and IL-1β in both THP-1 cells and PBMCs.In THP-1 cells,the suppression was partially caused by blockade of the translocation of NF-κB from the cytoplasm to the nucleus via affecting the TAK1-IKK-NF-κB pathway and p38 and ERK MAPK activity.Conclusion:Lobolide is a potential inhibitor of the NF-kB pathway,which blocks the translocation of NF-kB from the cytoplasm to the nucleus.Lobolide inhibits LPS-stimulated TNFα and IL-1β release,suggesting that the compound might be an anti-inflammatory compound.

  8. Kaurene diterpene induces apoptosis in U87 human malignant glioblastoma cells by suppression of anti-apoptotic signals and activation of cysteine proteases

    Directory of Open Access Journals (Sweden)

    F.S. Lizarte Neto

    2013-01-01

    Full Text Available Gliomas are the most common and malignant primary brain tumors in humans. Studies have shown that classes of kaurene diterpene have anti-tumor activity related to their ability to induce apoptosis. We investigated the response of the human glioblastoma cell line U87 to treatment with ent-kaur-16-en-19-oic acid (kaurenoic acid, KA. We analyzed cell survival and the induction of apoptosis using flow cytometry and annexin V staining. Additionally, the expression of anti-apoptotic (c-FLIP and miR-21 and apoptotic (Fas, caspase-3 and caspase-8 genes was analyzed by relative quantification (real-time PCR of mRNA levels in U87 cells that were either untreated or treated with KA (30, 50, or 70 µM for 24, 48, and 72 h. U87 cells treated with KA demonstrated reduced viability, and an increase in annexin V- and annexin V/PI-positive cells was observed. The percentage of apoptotic cells was 9% for control cells, 26% for cells submitted to 48 h of treatment with 50 µM KA, and 31% for cells submitted to 48 h of treatment with 70 µM KA. Similarly, in U87 cells treated with KA for 48 h, we observed an increase in the expression of apoptotic genes (caspase-8, -3 and a decrease in the expression of anti-apoptotic genes (miR-21 and c-FLIP. KA possesses several interesting properties and induces apoptosis through a unique mechanism. Further experiments will be necessary to determine if KA may be used as a lead compound for the development of new chemotherapeutic drugs for the treatment of primary brain tumors.

  9. Using the knowns to discover the unknowns: MS-based dereplication uncovers structural diversity in 17-hydroxygeranyllinalool diterpene glycoside production in the Solanaceae.

    Science.gov (United States)

    Heiling, Sven; Khanal, Santosh; Barsch, Aiko; Zurek, Gabriela; Baldwin, Ian T; Gaquerel, Emmanuel

    2016-02-01

    Exploring the diversity of plant secondary metabolism requires efficient methods to obtain sufficient structural insights to discriminate previously known from unknown metabolites. De novo structure elucidation and confirmation of known metabolites (dereplication) remain a major bottleneck for mass spectrometry-based metabolomic workflows, and few systematic dereplication strategies have been developed for the analysis of entire compound classes across plant families, partly due to the complexity of plant metabolic profiles that complicates cross-species comparisons. 17-hydroxygeranyllinalool diterpene glycosides (HGL-DTGs) are abundant defensive secondary metabolites whose malonyl and glycosyl decorations are induced by jasmonate signaling in the ecological model plant Nicotiana attenuata. The multiple labile glycosidic bonds of HGL-DTGs result in extensive in-source fragmentation (IS-CID) during ionization. To reconstruct these IS-CID clusters from profiling data and identify precursor ions, we applied a deconvolution algorithm and created an MS/MS library from positive-ion spectra of purified HGL-DTGs. From this library, 251 non-redundant fragments were annotated, and a workflow to characterize leaf, flower and fruit extracts of 35 solanaceous species was established. These analyses predicted 105 novel HGL-DTGs that were restricted to Nicotiana, Capsicum and Lycium species. Interestingly, malonylation is a highly conserved step in HGL-DTG metabolism, but is differentially affected by jasmonate signaling among Nicotiana species. This MS-based workflow is readily applicable for cross-species re-identification/annotation of other compound classes with sufficient fragmentation knowledge, and therefore has the potential to support hypotheses regarding secondary metabolism diversification.

  10. Lobolide, a diterpene, blockades the NF-κB pathway and p38 and ERK MAPK activity in macrophages in vitro

    Science.gov (United States)

    Lv, Xiao-fen; Chen, Si-han; Li, Jie; Fang, Jian-ping; Guo, Yue-wei; Ding, Kan

    2012-01-01

    Aim: Recent studies have shown that constitutive activation of the nuclear factor κB (NF-κB) plays a key role in chronic inflammation and cancers. The aim of this study was to characterize lobolide, a cembrane diterpene, as a drug candidate targeting the NF-κB signaling pathway. Methods: A HEK 293/NF-κB-Luc stable cell line was constructed to evaluate the effect of lobolide on NF-κB activation. THP-1 human monocytes and peripheral blood mononuclear cells (PBMCs) from healthy volunteers were tested. Lipopolysaccharide (LPS)-induced TNFα and IL-1β production and activation of the TAK1-IKK-NF-κB pathway were studied using ELISA and Western blot analysis. Results: In HEK 293/NF-κB-Luc stable cells, lobolide (0.19–50 μmol/L) inhibited NF-κB activation in a concentration-dependent manner with an IC50 value of 4.2±0.3 μmol/L. Treatment with lobolide (2.5–10 μmol/L) significantly suppressed LPS-induced production of TNFα and IL-1β in both THP-1 cells and PBMCs. In THP-1 cells, the suppression was partially caused by blockade of the translocation of NF-κB from the cytoplasm to the nucleus via affecting the TAK1-IKK-NF-κB pathway and p38 and ERK MAPK activity. Conclusion: Lobolide is a potential inhibitor of the NF-κB pathway, which blocks the translocation of NF-κB from the cytoplasm to the nucleus. Lobolide inhibits LPS-stimulated TNFα and IL-1β release, suggesting that the compound might be an anti-inflammatory compound. PMID:22922340

  11. Cubitene: an irregular twelve-membered-ring diterpene from a termite soldier. [1,5-dimethyl-8,10-bis(isopropenyl)cyclododeca-1,5-diene

    Energy Technology Data Exchange (ETDEWEB)

    Prestwich, G.D. (International Centre of Insect Physiology and Ecology, Nairobi, Kenya); Wiemer, D.F.; Meinwald, J.; Clardy, J.

    1978-04-12

    Cubitene was isolated from the hexane extract of C. umbratus soldier heads by chromatography over Florisil followed by preparative GLC. High resolution mass spectroscopy indicated the formula of cubitene to be C/sub 20/H/sub 32/. Mass spectra of catalytically hydrogenated cubitene products showed peaks at m/rho 279, consistent with their formulation as substituted cycloalkanes of the composition C/sub 20/H/sub 40/. Cubitene is therefore a monocyclic hydrocarbon with four centers of unsaturation. the /sup 1/H NMR spectral data revealed the presence of two 1,1-disubstituted double bonds, and two additional olefinic protons. Four methyl groups attached to double bonds are also observed. The /sup 13/C NMR spectrum indicated the presence of two tribsubstituted double bonds. Fortunately, cubitene could be obtained in crystalline form (mp 34.5-35/sup 0/C) from cold methanol, allowing a single-crystal x-ray diffraction analysis. It crystallized in the monoclinic crystal class. Lattice parameters were: a = 15.963 (5), b = 6.799 (2), c = 17.038 (5) A; theta = 96.99 (2)/sup 0/. Evidence shows cubitene to be 1,5-dimethyl-8,10-bis(isopropenyl)cyclododeca-1,5-diene, a structure in which one isoprene unit is irregularly joined to three others. It appears to be the first example of a diterpene hydrocarbon based on a twelve-membered carbocyclic ring. Its biosynthesis poses interesting problems. Two possible biosynthetic routes are suggested. An even more basic question remains....whether termite soldiers synthesize cubitene at all, or whether they simply sequester it (or a closely related precursor) from their food.

  12. Kaurene diterpene induces apoptosis in U87 human malignant glioblastoma cells by suppression of anti-apoptotic signals and activation of cysteine proteases.

    Science.gov (United States)

    Lizarte Neto, F S; Tirapelli, D P C; Ambrosio, S R; Tirapelli, C R; Oliveira, F M; Novais, P C; Peria, F M; Oliveira, H F; Carlotti Junior, C G; Tirapelli, L F

    2013-01-01

    Gliomas are the most common and malignant primary brain tumors in humans. Studies have shown that classes of kaurene diterpene have anti-tumor activity related to their ability to induce apoptosis. We investigated the response of the human glioblastoma cell line U87 to treatment with ent-kaur-16-en-19-oic acid (kaurenoic acid, KA). We analyzed cell survival and the induction of apoptosis using flow cytometry and annexin V staining. Additionally, the expression of anti-apoptotic (c-FLIP and miR-21) and apoptotic (Fas, caspase-3 and caspase-8) genes was analyzed by relative quantification (real-time PCR) of mRNA levels in U87 cells that were either untreated or treated with KA (30, 50, or 70 µM) for 24, 48, and 72 h. U87 cells treated with KA demonstrated reduced viability, and an increase in annexin V- and annexin V/PI-positive cells was observed. The percentage of apoptotic cells was 9% for control cells, 26% for cells submitted to 48 h of treatment with 50 µM KA, and 31% for cells submitted to 48 h of treatment with 70 µM KA. Similarly, in U87 cells treated with KA for 48 h, we observed an increase in the expression of apoptotic genes (caspase-8, -3) and a decrease in the expression of anti-apoptotic genes (miR-21 and c-FLIP). KA possesses several interesting properties and induces apoptosis through a unique mechanism. Further experiments will be necessary to determine if KA may be used as a lead compound for the development of new chemotherapeutic drugs for the treatment of primary brain tumors.

  13. Structure of the ent-Copalyl Diphosphate Synthase PtmT2 from Streptomyces platensis CB00739, a Bacterial Type II Diterpene Synthase.

    Science.gov (United States)

    Rudolf, Jeffrey D; Dong, Liao-Bin; Cao, Hongnan; Hatzos-Skintges, Catherine; Osipiuk, Jerzy; Endres, Michael; Chang, Chin-Yuan; Ma, Ming; Babnigg, Gyorgy; Joachimiak, Andrzej; Phillips, George N; Shen, Ben

    2016-08-31

    Terpenoids are the largest and most structurally diverse family of natural products found in nature, yet their presence in bacteria is underappreciated. The carbon skeletons of terpenoids are generated through carbocation-dependent cyclization cascades catalyzed by terpene synthases (TSs). Type I and type II TSs initiate cyclization via diphosphate ionization and protonation, respectively, and protein structures of both types are known. Most plant diterpene synthases (DTSs) possess three α-helical domains (αβγ), which are thought to have arisen from the fusion of discrete, ancestral bacterial type I TSs (α) and type II TSs (βγ). Type II DTSs of bacterial origin, of which there are no structurally characterized members, are a missing piece in the structural evolution of TSs. Here, we report the first crystal structure of a type II DTS from bacteria. PtmT2 from Streptomyces platensis CB00739 was verified as an ent-copalyl diphosphate synthase involved in the biosynthesis of platensimycin and platencin. The crystal structure of PtmT2 was solved at a resolution of 1.80 Å, and docking studies suggest the catalytically active conformation of geranylgeranyl diphosphate (GGPP). Site-directed mutagenesis confirmed residues involved in binding the diphosphate moiety of GGPP and identified DxxxxE as a potential Mg(2+)-binding motif for type II DTSs of bacterial origin. Finally, both the shape and physicochemical properties of the active sites are responsible for determining specific catalytic outcomes of TSs. The structure of PtmT2 fundamentally advances the knowledge of bacterial TSs, their mechanisms, and their role in the evolution of TSs.

  14. Proliferation inhibition, cell cycle arrest and apoptosis induced in HL-60 HL-60HL-60 HL-60 cells by a natural diterpene ester from Daphne mucronata

    Directory of Open Access Journals (Sweden)

    R Yazdanparast

    2011-05-01

    Full Text Available "n  "nBackground and the purpose of the study: Gnidilatimonoein (Gn, a new diterpene ester from Daphne mucronata, possesses strong anti-metastasis and anti-tumor activities. In this study, its apoptosis and differentiation capabilities were evaluated by using the leukemia HL-60 cell line. "nMaterial and methods: Cell prolifaration inhibition was estimated by MTT assay. The occurrence of apoptosis was evaluated by EtBr/AO double staining technique, cell cycle analyses and detection of apoptotic cells by Annexin V-FITC and propodium iodide (PI. Differentiation of the cells was determined by NBT reduction assay and the expression of specific cell surface markers such as CD14 and CD11b, were analyzed by flow cytometry.   "nResults: The drug decreased the growth of the cells dose- and time- dependently and the IC50 was found to be 1.3 µM. Our data suggested that Gn induced both monocytic differentiation  and apoptosis among HL-60 cells. In addition, cell cycle analyses showed an increase in G1 phase population by 24 hrs, which was gradually replaced by Sub-G1 cell population (apoptotic cells by 72 hrs. "nConclusion: Based on these data, the Gn-treated HL-60 cells displayed differentiation-dependent apoptosis. Thus, Gn might be a good candidate for differentiation therapy of leukemia, pending full biological evaluation of the compound among the wide array of leukemia cells. "nevaluation of the compound among the wide array of leukemia cells.

  15. Propostas para o estudo dos protestantismos ibéricos

    OpenAIRE

    Afonso, José António

    2004-01-01

    A propósito da edição da obra “Las minorias religiosas en España y Portugal: passado y presente”, número monográfico da revista “Anales de Historia Contemporanea” (Universidade de Múrcia), o Autor elenca algumas propostas para o estudo dos protestantismos ibéricos.

  16. ACREDITAÇÃO HOSPITALAR: UM ESTUDO DE CASO

    Directory of Open Access Journals (Sweden)

    Felipe Antunes

    2005-06-01

    Full Text Available Este artigo aborda a qualidade e acreditação hospitalar através do Manual das Organizações Prestadoras de Serviços Hospitalares, desenvolvido pela Organização Nacional de Acreditação (ONA. O objetivo geral deste trabalho é avaliar o processo de acreditação, identificando dificuldades e benefícios advindos da implantação deste  Manual. Após a realização de uma revisão bibliográfica sobre qualidade em hospitais e normas da qualidade aplicáveis a hospitais, é feita uma análise crítica teórica do Manual das Organizações Prestadoras de Serviços Hospitalares. Tal análise crítica inclui um estudo comparativo entre os requisitos do Manual e da norma NBR ISO 9001/2000. A seguir, a análise teórica é complementada através de um estudo de caso contemplando a implantação do Manual junto a um hospital de grande porte. Finalmente, são apresentadas as conclusões, baseadas tanto no estudo teórico como no estudo prático.

  17. Pragmatismo e os pragmáticos nos estudos organizacionais

    Directory of Open Access Journals (Sweden)

    Francis Kanashiro Meneghetti

    Full Text Available O objetivo central deste ensaio teórico é avaliar as contribuições do pragmatismo nos Estudos Organizacionais e, os específicos, avaliar se as contribuições teóricas de Charles Peirce e John Dewey possibilitam afirmar o pragmatismo como senso comum, ciência ou teoria do conhecimento; analisar as contribuições - possibilidadese limites - do pragmatismo como conhecimento científico; compreender os percursos do pragmatismo nos Estudos Organizacionais. Chaga-se a conclusão de que ser pragmático é diferente de conceber estudos articulados no campo epistemológico do pragmatismo. O sujeito pragmático vê na sua utilidade e na necessidade deação, o ponto de tensão que coloca em dúvida a veracidade dos pressupostos do pragmatismo como ciência. O que ocorre nos Estudos Organizacionais éa conversão do pragmatismo como conhecimento científico emsubordinação irracional à utilidade e à ação prática, sobretudo, no campo da economia, da política, da ciência, da cultura, da educação, e assim por diante.

  18. Research progress on metabolic mechanism of cembranoid diterpenes and its regulation%类西柏烷二萜代谢机理及调控研究进展

    Institute of Scientific and Technical Information of China (English)

    王冬; 张小全; 杨铁钊; 薛刚; 李丽华

    2014-01-01

    综述了类西柏烷二萜合成的生化途径及其调控因素,并阐述了其在改善作物香味和植物抗虫方面的作用及研究利用前景。类西柏烷二萜主要存在于烟草属植物的腺毛分泌物中,在西柏三烯醇合酶(CBTS)和细胞色素P450加氧酶(CYP450)的催化下经脱氧木酮糖磷酸代谢途径(DXP)合成,这一过程受到基因和多种外界因素的调控。%Biosynthetic and metabolic pathway and regulatory factors of cembranoid diterpenes were introduced, and its application prospective on aroma-improving and insect-resistant were also discussed. Cembranoid diterpenes mainly exists in glandular trichome secretion of Nicotiana plant. It was produced through plastidic deoxyxylulose phosphate (DXP) pathway with cembratrien-ol synthase (CBTS) and cytochrome P450 oxygenase (CYP450) as catalyzer. This process was regulated by gene and various external factors.

  19. Contribuição para o estudo da versão portuguesa da Positive and Negative Affect Schedule (PANAS): II – Estudo psicométrico

    OpenAIRE

    2005-01-01

    Estudo da adaptação da Positive and Negative Affect Schedule (PANAS; Watson, Clark & Tellegen, 1988) para a população portuguesa. A metodologia do estudo procurou replicar o estudo de desenvolvimento da escala original. Tal como o estudo original, pediu-se a 348 estudantes universitários que respondessem em que medida experienciaram as 60 emoções (traduzidas para Português) propostas por Zevon e Tellegen (1982) “durante os últimos dias” e “durante as últimas semanas”, numa escala de 5 pontos....

  20. Estudos da formação de planetas terrestres

    OpenAIRE

    Costa, André Izidoro Ferreira da [UNESP

    2013-01-01

    O estudo da formação de planetas terrestres no Sistema Solar, é crucial para compre- endermos como outros sistemas planetários formam e também inferir as condições que poderiam ter influenciado a origem e evolução de vida na Terra. Esta Tese de douto- rado apresenta um estudo numérico da formação de planetas terrestres. Nosso objetivo principal é analisar o último estágio da formação desses planetas no Sistema Solar, em particular, a formação de Marte e a origem da água da Terra. Esses dois p...