Didier Stien

Antimalarial Activity of Simalikalactone E, a New Quassinoid from Quassia amara L. (Simaroubaceae)

ABSTRACT We report the isolation and identification of a new quassinoid named simalikalactone E (SkE), extracted from a widely used Amazonian antimalarial remedy made out of Quassia amara L. (Simaroubaceae) leaves. This new molecule inhibited the growth of Plasmodium falciparum cultured in vitro by 50%, in the concentration range from 24 to 68 nM, independently of the strain sensitivity to chloroquine. We also showed that this compound was able to decrease gametocytemia with a 50% inhibitory concentration sevenfold lower than that of primaquine. SkE was found to be less toxic than simalikalactone D (SkD), another antimalarial quassinoid from Q. amara, and its cytotoxicity on mammalian cells was dependent on the cell line, displaying a good selectivity index when tested on nontumorogenic cells. In vivo, SkE inhibited murine malaria growth of Plasmodium vinckei petteri by 50% at 1 and 0.5 mg/kg of body weight/day, by the oral or intraperitoneal routes, respectively. The contribution of quassinoids as a source of antimalarial molecules needs therefore to be reconsidered.

Diversity of the Volatile Organic Compounds Emitted by 55 Species of Tropical Trees: a Survey in French Guiana

Volatile organic compounds (VOCs) are produced by a broad range of organisms, from bacteria to mammals, and they represent a vast chemical diversity. In plants, one of the preeminent roles of VOCs is their repellent or cytotoxic activity, which helps the plant deter its predators. Most studies on VOCs emitted by vegetative parts have been conducted in model plant species, and little is known about patterns of VOC emissions in diverse plant communities. We conducted a survey of the VOCs released immediately after mechanical damage of the bark and the leaves of 195 individual trees belonging to 55 tropical tree species in a lowland rainforest of French Guiana. We discovered a remarkably high chemical diversity, with 264 distinct VOCs and a mean of 37 compounds per species. Two monoterpenes (α-pinene and limonene) and two sesquiterpenes (β-caryophyllene and α-copaene), which are known to have cytotoxic and deterrent effects, were the most frequent compounds in the sampled species. As has been established for floral scents, the blend of VOCs is largely species-specific and could be used to discriminate among 43 of the 55 sampled species. The species with the most diverse blends were found in the Sapindales, Laurales, and Magnoliales, indicating that VOC diversity is not uniformly distributed among tropical species. Interspecific variation in chemical diversity was caused mostly by variation in sesquiterpenes. This study emphasizes three aspects of VOC emission by tropical tree species: the species-specificity of the mixtures, the importance of sesquiterpenes, and the wide-ranging complexity of the mixtures.

Search for antifungal compounds from the wood of durable tropical trees.

Research on antifungal compounds from the durable wood from French Guiana Amazonian forest trees highlights the correlation between the activity of their extracts against wood-rotting fungi and human pathogens. The fractionation of an ethyl acetate extract of Sextonia rubra wood led to the isolation of rubrenolide (1) and rubrynolide (2). The potential of compounds 1 and 2 is described through the evaluation of their activity against 16 pathogenic fungi and their cytotoxicity toward NIH-3T3 mammalian fibroblast cells.

Antileishmanial sesquiterpene lactones from Pseudelephantopus spicatus, a traditional remedy from the Chayahuita Amerindians (Peru). Part III

ETHNOPHARMACOLOGICAL RELEVANCEnThe study of traditional remedies used by the Chayahuita, an ethnic group from the Peruvian Amazonia, has prompted us to investigate in detail the ethanolic extract of Pseudelephantopus spicatus (Juss. ex Aubl.) C.F. Baker, which has demonstrated strong biological activity towards Leishmania amazonensis. Our goal was to discover the active compound of this plant-based remedy.nnnMATERIALS AND METHODSnA bioguided fractionation of the crude extract was undertaken based on the biological activity recorded against Leishmania amazonensis axenic amastigotes in in vitro bioassays.nnnRESULTSnThree strongly to moderately active compounds were isolated: two hirsutinolides (the 8,13-diacetyl-piptocarphol and the 8-acetyl-13-O-ethyl-piptocarphol) and ursolic acid. IC(50) against Leishmania amazonensis axenic amastigotes are respectively 0.2, 0.37 and 0.99 μM (while IC(50) of amphotericin B is 0.41 μM). These compounds have never been isolated from this plant species, and germacranolides have never been identified as potential antileishmanial agents.nnnCONCLUSIONSnThe compounds isolated from Pseudelephantopus spicatus account for the antileishmanial activity of the plant, thus giving support to its use by the Chayahuita in Peru.

Treatment of leishmaniasis in the Oyapock basin (French Guiana): A K.A.P. survey and analysis of the evolution of phytotherapy knowledge amongst Wayãpi Indians

ETHNOPHARMACOLOGICAL RELEVANCEnCutaneous leishmaniasis is a neglected disease with a high incidence in French Guiana, mainly in the middle and upper Oyapock basin, where Amerindian and some Brazilian people live. The main goals of this work were (i) to assess the knowledge about leishmaniasis in the different populations of the middle and upper Oyapock basin, (ii) to study the therapeutic strategies adopted by people affected by leishmaniasis and (iii) to document the use of phytotherapeutic remedies for leishmaniasis. Knowledge, attitudes and practices (K.A.P.) related to this disease and its treatments have been studied according to cultural group and geographical settlement. Within the Wayãpi group, the evolution of the knowledge of phytoremedies over the last 20 years has been characterised by literature-based comparisons.nnnMATERIALS AND METHODSnA total of 144 questionnaires were administered in all the villages of the upper Oyapock and Camopi basins. Correspondence analyses were used for multivariate analysis. Plant species were identified at the Cayenne Herbarium (CAY).nnnRESULTSnThe biomedical concept of leishmaniasis correlates well with the Teko and Wayãpi concepts of kalasapa and kalasapau. Although the vector of this disease was not correctly identified, the most commonly cited aetiology (74.5%) was vector-borne, and related epidemiological schemes correlate well with the one encountered in French Guiana. Theoretically and practically, health centres were the most commonly used resource for diagnostic in instances of leishmaniasis infection (65.9%), independently of the patients cultural group, along with the use of pharmaceutical drugs (85.3%). Pharmaceuticals were commonly utilised despite the frequent (51.5%) use of phytotherapeutic remedies, alone or in combination with drugs. The most cited medicinal plant species for the treatment of leishmaniasis included Eleutherine bulbosa (Mill.) Urb. (Iridaceae, cited 14 times), Euterpe oleracea Mart. (Arecaceae, 9), Cecropia obtusa Trecul (Cecropiaceae, 8), Jatropha curcas L. (Euphorbiaceae, 7), Ceiba pentandra (L.) Gaertn. (Bombacaceae, 6) and Carica papaya L. (Caricaceae, 6). Multiple correspondence analyses demonstrated that the species used in leishmaniasis remedies are more prone to vary by the users place of residence than by their cultural origin, which indicates that exchange of knowledge about leishmaniasis remedies has occurred across different cultural groups. Literature-based comparisons between the remedies for leishmaniasis used by the Wayãpi during the 1980s showed a striking evolution, both in terms of diversity of species and number of plants used. The large number of species shared with other Guianese groups argues for intercultural exchange and may explain the majority (57.1%) of the newly used species highlighted in our study.nnnCONCLUSIONSnLeishmaniasis is a well-known disease in the studied area. Phytotherapeutic treatments are still in use, although they are not the main source of remedies, and should undergo pharmacological studies to evaluate their potential therapeutic value.

Quassinoid constituents of Quassia amara L. leaf herbal tea. Impact on its antimalarial activity and cytotoxicity

AIM OF THE STUDYnOur objective was to assess whether it could be contemplated to recommend Quassia amara young leaf tea for treatment against malaria, and if yes, set up a standard protocol for preparing the herbal tea.nnnMATERIALS AND METHODSnThe leaf tea was extracted with methylene chloride and the organic extract was fractionated with HPLC. Pure compounds were characterized and their in vitro cytotoxicity and antiplasmodial activity was determined.nnnRESULTS AND DISCUSSIONnWe discovered that antimalarial Quassia amara young leaf tea contains several quassinoids: simalikalactone D (SkD, 1), picrasin B (2), picrasin H (3), neoquassin (4), quassin (5), picrasin I (6) and picrasin J (7). These last two compounds are new. In addition, our experiments demonstrate that both biological activity and cytotoxicity of the remedy may be attributed solely to the presence of SkD.nnnCONCLUSIONnIn conclusion, this preparation should not be recommended for treatment of malaria until a clinical study in humans is performed with SkD.

Secondary metabolites of Bagassa guianensis Aubl. wood: A study of the chemotaxonomy of the Moraceae family

In order to explain the durability of the Moraceae plant family, phytochemistry of Bagassa guianensis was performed. Ethyl acetate extract was obtained from the heartwood and 18 secondary metabolites were isolated, including 6 moracins [6-O-methyl-moracin M, 6-O-methyl-moracin N and moracin Z; previously identified: moracin M, moracin N and moracin P], 8 stilbenoids [presently identified: (-)-epialboctalol and arachidin 4; previously identified: alboctalol, trans-resveratrol, arachidin 2, trans-oxyresveratrol and artogomezianol], 3 previously identified flavonoids, steppogenin, katuranin and dihydromorin, beta-sitosterol and resorcinol. Previous studies suggest that stilbenoids are responsible for the natural durability of wood. Our study has determined that B. guianensis is closely related to Morus sp. in phylogeny and should be included in the Moreae sensu stricto tribe of the Moraceae family.

Medical ethnobotany of the Chayahuita of the Paranapura basin (Peruvian Amazon)

ETHNOPHARMACOLOGICAL RELEVANCEnUp until now, the plant pharmacopoeia of the Chayahuita, an ethnic group from the Peruvian Amazon, has been poorly defined. This paper details the uses of medicinal plants within this community, as recorded in two villages of the Paranapura basin, Soledad and Atahualpa de Conchiyacu. This study aimed to describe the basis of the Chayahuita traditional medical system, to document part of the medicinal plant corpus, and to compare it with data from other Amazonian ethnic groups.nnnMATERIAL AND METHODSnMethodology was based (i) on field prospection with 26 informants (ethnobotanical walks methodology), (ii) semi-structured interviews including 93 people (49 men and 44 women) focused on the most recent health problem experienced and on the therapeutic options chosen, (iii) individual or group thematic discussions relating to disease and treatments, (iv) 6-months of participants observations between May 2007 and May 2008. At the end of the project in May 2008 a workshop was organized to cross-check the data with the help of 12 of the most interested informants.nnnRESULTSnSix hundred and seventeen voucher specimens were collected, corresponding to 303 different species, from which 274 (belonging to 83 families) are documented here. Altogether 492 recipes were recorded, corresponding to a global figure of 541 therapeutic uses and a total of 664 use reports. The main therapeutic uses are related to dermatological problems (103 uses; 19%), gastro-intestinal complaints (69 uses; 13%) and malaria/fevers (52 uses; 10%). Diseases are analysed according to Chayahuita concepts, and for each disease the species having a high frequency of citation are listed, and the most frequently used remedies are described. Whenever possible, comparisons with other Amazonian groups have been drawn.nnnCONCLUSIONnChayahuita nosology and medical ethnobotany appear to draw their inspiration from a common panamazonian root. Despite the fact that a certain number of medicinal plants are shared with other nearby groups, there seem to be specific uses for some species, thus highlighting the originality of the Chayahuita pharmacopoeia. Presently there is a certain disinterest in the most traditional area of the Chayahuita medical ways, and the role of the penutu (shaman) seems to be less highly-valued than in the past. Nonetheless, the use of medicinal plants in phytotherapeutic treatment is very much a living, shared knowledge.

Ta'ta', Huayani: perception of leishmaniasis and evaluation of medicinal plants used by the Chayahuita in Peru. Part II.

AIM OF THE STUDYnA knowledge attitude and practice study centred on leishmaniasis and its treatment was performed among the Chayahuita, an Amazonian Peruvian ethnic group living in an endemic area. This study documents traditional Chayahuita plants use and disease concepts. Also, activity of some medicinal plants used by the Chayahuita is highlighted and discussed.nnnMATERIALS AND METHODSnNinety-three Chayahuita people were interviewed, following a semi-structured questionnaire focussed on disease knowledge and perception, personal attitude and healing practices. Simultaneously, a collection of plants was performed in different ecotopes, in order to make an extensive inventory of the pharmacopoeia.nnnRESULTSnFor the Chayahuita, cutaneous (CL) and muco-cutaneous leishmaniasis (MCL) are considered as diseases of their own, with specific names, aetiologies, mode of transmission. Regarding CL, Chayahuita people consider that the humid characteristic of the skin ulcer is a discriminative fact orienting the diagnostic forTata (leishmaniasis). Forty-six different species were designated useful against LC and /or MCL (29 species by means of the questionnaire and 27 species when collecting in different ecotopes). Thirty-seven extracts corresponding to 31 species used medicinally were screened in vitro against Leishmania amazonensis axenic amastigotes, assessing their viability by the reduction of tetrazolium salt (MTT). Six species displayed a good activity (10 microg/ml<IC50<20 microg/ml): an undetermined hemi-epiphytic Clusiaceae, Cybianthus anthuriophyllus Pipoly (Myrsinaceae), two Piper, Piper sanguineispicum Trel., and Piper loretoanum Trel. (Piperaceae), Desmodium axillare Sw. DC. (Fabaceae), and Clibadium sylvestre (Aubl.) Baill. (Asteraceae).nnnCONCLUSIONnPerception of leishmaniasis, attitude, treatments and diet prohibitions still largely reflects traditional Chayahuita cosmovision, even if some tentative of bio-medical re-interpretation is arising.

Secondary metabolites from Spirotropis longifolia (DC) Baill and their antifungal activity against human pathogenic fungi.

A phytochemical study of the ethyl acetate extract of the roots and adventitious roots of Spirotropis longifolia, a monodominant tree species of the Guianan rainforest, has allowed the isolation of three compounds: 2-hydroxy-8,9-methylenedioxy-2,2-dimethylpyrano-[5,6:4,3]-6a-prenyl-[6aS,11aS]-pterocarpan (spirotropin A), 2-hydroxy-8,9-methylenedioxy-2,2-dimethyl-3,4-dihydropyrano-[5,6:4,3]-6a-prenyl-[6aS,11aS]-pterocarpan (spirotropin B), and 5,7-dihydroxy-6,8-diprenyl-2,2-dimethylpyrano[5,6: 3,4]-isoflavone (spirotropone). In addition, 10 known compounds, trans-oxyresveratrol, trans-resveratrol, piceatannol, daidzein, genistein, isoprunetin, lupeol, latifolol, gnetin D and gnetin E, were also isolated. These compounds were evaluated for their antifungal activity and their cytotoxicity, and their structures were established by 1D and 2D NMR, HRMS, CD and optical rotation measurements.