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Annona senegalensis Pers. is a plant used in Burkina folkloric medicine for the treatment of epilepsy. The stem-bark, roots and leaves of A. senegalensis are used for an array of human disorders, including epilepsy. Previous studies have demonstrated some genuine effects of A. senegalensis fractions in vivo models of temporal lobe epilepsy. In the present work, we have compared the pharmacological profile of the most promising A. senegalensis fraction obtained up-to-date with anticonvulsant drugs targeting voltage-activated channels namely Carbamazepine (Na+ channels) and Retigabine (KCNQ Channels). In an in vitro model of temporal lobe epilepsy, we could observe a dose-dependent reduction of electrically evoked epileptiform bursts in the CA1 region of the rat hippocampus when applying carbamazepine, retigabine or the F4 fraction of A. senegalensis. The intrinsic excitability of CA1 pyramidal neurons was dose-dependently reduced by A. senegalensis. Since A. senegalensis effects could not anymore be observed when endogenous voltage-dependent sodium channels were blocked and intrinsic excitability restored with a dynamic clamp amplifier, it was concluded that A. senegalensis contains molecules that target voltage-dependent sodium channels. Key words: Hippocampus, epilepsy, Annona senegalensis, Burkina Faso plant extracts, sodium channel, KCNQ channel, M-current, retigabine, carbamazepine.

The activity of β-caryophyllene oxide as either a contact or noncontact repellent was evaluated against two laboratory strains (Aedes albopictus and Anopheles dirus) using an excito-repellency test system. N, N-Diethyl-3-methylbenzamide (DEET) was used as a standard reference baseline for comparative purposes. β-Caryophyllene oxide and DEET were tested at concentrations of 0.1, 0.25, 0.5 and 1.0% (v/v). In addition, the phototoxic and genotoxic effects of β-caryophyllene oxide were investigated on Balb/c 3T3 mouse fibroblasts (3T3-L1) and Chinese hamster ovary cell line (CHO-K1). The results demonstrated that the higher concentrations of test compounds (0.5 and 1.0%) produced greater behavioral responses. Aedes albopictus was more sensitive to β-caryophyllene oxide than An. dirus. Moderate avoidance response rates (25-56% escape) of Ae. albopictus at 0.5% and 1.0% β-caryophyllene oxide were observed in contact and noncontact trials compared with low response rates from An. dirus (26-31% escape). DEET at ≤1% displayed lower irritancy and repellency (1-38%) than β-caryophyllene oxide when tested against the two mosquito species. Knockdown responses (37%) were only observed in An. dirus exposed to 1% β-caryophyllene oxide in the contact trial. β-Caryophyllene oxide did not show any phototoxic potential (PIF= 0.38) nor was there any significant genotoxic response as indicated by no increase in micro-nucleated cells with or without metabolic activation. β-Caryophyllene oxide could be considered as a safe repellent, effective against mosquitoes.

Six limonoids [kotschyienone A and B (1, 2), 7-deacetylgedunin (3), 7-deacetyl-7-oxogedunin (4), andirobin (5) and methyl angolensate (6)] were investigated for their trypanocidal and leishmanicidal activities using bloodstream forms of Trypanosoma brucei and promastigotes of Leishmania major. Whereas all compounds showed anti-trypanosomal activity, only compounds 1–4 displayed anti-leishmanial activity. The 50% growth inhibition (GI50) values for the trypanocidal and leishmanicidal activity of the compounds ranged between 2.5 and 14.9 μM. Kotschyienone A (1) was found to be the most active compound with a minimal inhibition concentration (MIC) value of 10 μM and GI50 values between 2.5 and 2.9 μM. Only compounds 1 and 3 showed moderate cytotoxicity against HL-60 cells with MIC and GI50 values of 100 μM and 31.5–46.2 μM, respectively. Compound 1 was also found to show activity against intracellular amastigotes of L. major with a GI50 value of 1.5 μM. The results suggest that limonoids have potential as drug candidates for the development of new treatments against trypanosomiasis and leishmaniasis.

Background Many medicinal plants have been used in Vietnam but most of them have not been studied and documented. In this study, the traditional plants used by K’Ho-Cil people living in BiDoup-Nui Ba National Park, Vietnam for treatment of diarrhea and identified potential medicinal plants were documented. Methods Information of the medicinal plants used was collected through field surveys and semi-structured interviews with key informants. Plants used for anti-diarrhea were selected for further analysis in structured questionnaires. The potential medicinal plants were identified through quantitative tools: fidelity level (FL) and choice value (CV). The relationship between traditional knowledge and gender, age and geological site was also evaluated using statistical analysis. Results Of the 133 taxa reported by 12 key informants, 34 anti-diarrheal plants were confirmed by 127 non-specialist informants. Interestingly, these plants were also used for 35 other health problems. Among the medicinal plants, Asteraceae was the most predominant family. Elephantopus mollis and Chromolaena odorata belonging to this family with the highest CV were the most preferred ones in diarrheal treatment (1.23 and 1.04, respectively). Moreover, in regard to FL and literature review, several potential plants for further studies were identified, for example Hedyotis capitellata var pubescens and Syzygium ripicola (FL = 100%), or Clitoria mariana and Dacrycarpus imbricatus (FL = 80 %). Conclusion In Vietnam, folk medicine has been widely used by many people without documentation for preservation and scientific study. This study contributes a database of anti-diarrhea plants used by K’Ho-Cil people living in BiDoup-Nui Ba National Park, Vietnam.

Ethnopharmacological relevance Flowers of Inula montana L. (Asteraceae), commonly known as “Arnica de Provence”, are used in the traditional medicine of Provence in France with the same indication as Arnica montana, for the relief of bruises, as an anti-inflammatory agent. Aims of the study The aim of our study is to evaluate its anti-inflammatory properties and to justify its traditional uses. Its potential valorization is evaluated in order to propose Inula montana as an alternative to Arnica montana. Materials and methods Bio-guided fractionation of ethanolic extract allowed the isolation of compounds responsible of the inhibition of NO production. The fractionation was realized using chromatographic techniques and structure elucidation was conducted by ESI-MS and NMR spectral data. Anti-inflammatory effect of ethanolic extract, different fractions and isolated pure compounds was studied in vitro on immortalized mouse macrophages RAW 264.7. An analytical UHPLC-DAD-ESI-MS/MS method was developed for the identification of these compounds in the herbal drug. This UHPLC-DAD method was validated and was used to compare the phenolic profile and content in plant material from the two collection sites: Bonnieux and Merindol. Results Eleven compounds were identified by UHPLC-MS. Chlorogenic acid (1), Luteolin (2), Nepetin (3), 3,5-O-Dicaffeoylquinic acid (4), 1,5-O-Dicaffeoylquinic acid (5), Nepitrin (6), Hispiduloside (7) and Jaceosid (8) were isolated and identified by NMR. Compounds 9, 10 and 11 were confirmed to be 6-Hydroxykaempferol 3,7-dimethyl ether, Hispidulin and Chrysosplenol C, respectively by comparing retention times and MS/MS data with those of the authentic substances. Six compounds: 1 and 4–8 are reported for the first time in Inula montana L. Compounds 2–8 showed promising anti-inflammatory activity with the release of NO with IC50 value Conclusion Flowers extracts and isolated compounds present promising anti-inflammatory activity which provides a scientific basis for the traditional use of Inula montana and may be proposed in the same indications as Arnica montana. The developed and validated simple, accurate and rapid UHPLC method can be used for the quality control of the herbal drug.

Les goudrons végétaux sont des produits résineux néoformés extraits par pyrolyse du bois ou de l’écorce (cf. bouleau), contrairement aux goudrons d’origine géologique (bitume, goudron de houille ou de schiste). Le principe d’extraction est de faire exsuder la matière première en la soumettant à un traitement thermique direct ou indirect ; l’exsudat se transforme alors en goudron. En Europe et en Méditerranée, les exsudats étaient extraits de diverses familles végétales en fonction des époques...

An ultrasound-assisted extraction (UAE) was optimized for the extraction of three bioactive alkaloids, namely, palmatine, roemerine, and tetrahydropalmatine from the tuber of Stephania cambodica . The performance of UAE and microwave-assisted extraction (MAE) was compared with that of percolation. UAE and MAE are also more environmentally friendly and use less toxic chemicals. MAE was as efficient as UAE for the extraction of tetrahydropalmatine; however, UAE significantly enhanced extraction yields of palmatine and roemerine. The influence of effective variables viz. liquid–solid ratio, percentage of ethanol, and time on UAE was then investigated using response surface methodology. The optimal UAE conditions were found to be ethanol 52%, time 9 min, and liquid–solid ratio 26.6:1 mL/g. Structural change was conducted by scanning electron microscopy. These findings suggest that UAE is a practical extractive procedure for preparation of alkaloid-rich extract for pharmacological investigation and for analytical control of S. cambodica tuber.

A structure-activity relationship study of active molecules against chloroquine-resistant Plasmodium falciparum K1 strain is reported. Structurally simplified analogues of antiplasmodial active alkaloids presented similar levels of activity as their corresponding natural products extracted from Guiera senegalensis and Mitragyna inermis with IC50 values on chloroquine-resistant P. falciparum K1 strain of up to 10.6 μM for spirooxindoles and 13.8 μM for β-carbolines. The identification of such simpler and cheaper structural analogues is crucial to efficiently study these natural products' action mode as well as developing new cures against malaria.

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Les goudrons vegetaux sont des produits resineux neoformes extraits par pyrolyse du bois ou de l’ecorce (cf. bouleau), contrairement aux goudrons d’origine geologique (bitume, goudron de houille ou de schiste). Le principe d’extraction est de faire exsuder la matiere premiere en la soumettant a un traitement thermique direct ou indirect ; l’exsudat se transforme alors en goudron. En Europe et en Mediterranee, les exsudats etaient extraits de diverses familles vegetales en fonction des epoques...

The new acylated flavonoid bioside 4′-O-methylisoscutellarein-7-O-[4′′′-O-acetyl]allopyranosyl-(1→2)-glucopyranoside (1) was isolated from the aerial parts of Stachys annua L. (Lamiaceae). Subterranean organs yielded for the first time 4′-O-methylisoscutellarein (2) and 4′-O-methylisoscutellarein-7-O-[6′′-Oacetyl] allopyranosyl-(1→2)-glucopyranoside (3). Chemical structures of the isolated compounds were elucidated using NMR spectroscopy.

Furostanol and spirostanol glycosides 1 and 2 were isolated from the flowers of Allium leucanthum, a Caucasian endemic species that grows in Georgia. The structures were established on the base of chemical evidence and spectral analyses ( 1 H, 13 C NMR, 1 H– 1 H COSY, 1 H– 13 C COSY, HMBC, and HR-MS) data. Compound 1 (leucofuranoside A) was reported for the first time and was identified as 26-O-β-Dglucopyranosyl-(25R)-5α-furostane-3β,6β-diol-3-O-β-D-glucopyranosyl-(1→2)-O-β-D-xylopyranosyl-(1→3)-O-β-D-glucopyranosyl-(1→4)-β-D-galactopyranoside. Compound 2 was identified as (25R)-5α-spirostane-3β,6β-diol-3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl-(1→4)-β-D-galactopyranoside and described for the first time in the genus Allium.

International audience; Inula montana is a member of the family Asteraceae and is present in substantial numbers in Garrigue country (calcareous Mediterranean ecoregion). This species has traditionally been used for its anti-inflammatory properties as well as Arnica montana. In this study, three habitats within Luberon Park (southern France) were compared regarding their pedoclimatic parameters and the resulting morpho-physiological response of the plants. The data showed that I. montana grows in south-facing poor soils and tolerates large altitudinal and temperature gradients. The habitat conditions at high elevation appear to affect mostly the morphology of the plant (organ shortening). Although the leaf contents of total polyphenols and flavonoids subclass essentially followed a seasonal pattern, many sesquiterpene lactones were shown to accumulate first at the low-elevation growing sites that suffered drought stress (draining topsoil, higher temperatures and presence of a drought period during the summer). This work highlights the biological variability of I. montana related to the variation of its natural habitats which is promising for the future domestication of this plant. The manipulation of environmental factors during cultivation is of great interest due to its innovative perspective for modulating and exploiting the phytochemical production of I. montana.

International audience; Introduction: Drugs resistant Plasmodium falciparum is a recurring issue that threatens public health. New anti-plasmodial drugs are needed to overcome this problem. The aim of this study was to characterize in vitro an-tiplasmodial activity of Withania frutescens (Solanaceae) against chloroquine-resistant Plasmodium falciparum strain. Methods: The in vitro antiplasmodial activity of leaves and roots extracts from Withania frutescens was performed in 96 well plates and preliminary phytochemical analysis was performed for the active fractions. The toxicity of the plant extract against CHO (Chinese Hamster Ovary) cells was assessed using the tetrazolium salt MTT col-orimetric assay. The selectivity index (SI) was calculated as the ratio between the cytotoxic and antiparasitic activities. Results: The methanol extract of W. frutescens leaves showed a good antiplasmodial activity (CC 50 18.1 μg/ml). Furthermore, ethyl acetate and butanol fractions showed promising in vitro antiplasmodial activity with a se-lectivity index of 3.1 and 2.4, respectively. The roots of W. frutescens were found to be inactive with a CC 50 value > 80 μg/ml. Conclusion: The antiplasmodial activity of W. frutescens may in part be attributed to the presence of polyphenolic and flavonoid compounds. Based on our results, ethyl acetate and butanol leaves fractions could be considered as a promising source for the development of putative antiplasmodial drugs.

We provide in this article the catalogue of the species names currently accepted in the genus Stephania (Menispermaceae), together with information relative to their types. Sixty-nine species of Stephania are recognized and 36 lectotypes are designated here. This checklist constitutes the basis for a future taxonomical revision of the genus Stephania.

International audience; Fractionation of the ethyl acetate-soluble extract of the roots of Leplaea mayombensis afforded two new 3,4-seco-lanostane-type triterpenoids, leplaeric acids A and B (1, 2), the new lanostane-type triterpenoid leplaeric acid C (3), and six known natural products (5–10). Derivatization of the main constituent, 1, afforded the dimethyl ester 4, the monoamide 11, and diamide 12 for SAR studies. The structures of these compounds were established through spectroscopic methods, and a single-crystal X-ray diffraction analysis was used to confirm the relative configuration of compound 1. These lanostane derivatives are unique since they are the first C-21-oxygenated lanostanes isolated from plant sources. Preliminary biological assays against the MDA MB 231 breast cancer cell line showed that compounds 1, 2, 4, and 11 have modest cytotoxic activity. Compound 2 was the most active, with an IC50 of 55 ± 7 μM. From these results, the amides (11, 12) derived from triterpenoid 1 were found to be less active than the derived esters (2, 4).

International audience; A phytochemical investigation of the ethanol extract of leaves and flowers of Inula montana L. led to the isolation of one new sesquiterpene acid called Eldarin (1) and four new inositol derivatives, Myoinositol,1,5-diangelate-4,6-diacetate (2), Myoinositol,1,6-diangelate-4,5-diacetate (3), Myoinositol-1-angelate-4,5-diacetate-6-(2-methylbutirate) (4), Myoinositol-1-angelate-4,5-diacetate-6-isovalerate (5) isolated for the first time, along with eleven known compounds described for the first time in Inula montana, 1β-Hydroxyarbusculin A (6), Artemorin (7), Santamarin (8), Chrysosplenol C (9), 6-Hydroxykaempferol 3,7-dimethyl ether (10), Reynosin (11), 2 Calenduladiol-3-palmitate (12), Costunolide (13), 4-Hydroxy-3,5-dimethoxybenzenemethanol (14), 9β-Hydroxycostunolide (15) and Hispidulin (16). Structural elucidation has been carried out by spectral methods, such as 1D and 2D NMR, IR, UV and HR-ESI-MS. These compounds have been tested in vitro for anti-inflammatory and cytotoxic activity on macrophages RAW 264.7. As a result, compounds 2, 3, 7, 13, 14, 15 and 16 showed a release of NO with IC 50 value < 30 µM on macrophages.

Ethnopharmacological relevance Stephania rotunda Lour. (Menispermaceae) is an important traditional medicinal plant that is grown in Southeast Asia. The stems, leaves, and tubers have been used in the Cambodian, Lao, Indian and Vietnamese folk medicine systems for years to treat a wide range of ailments, including asthma, headache, fever, and diarrhoea. Aim of the review: To provide an up-to-date, comprehensive overview and analysis of the ethnobotany, phytochemistry, and pharmacology of Stephania rotunda for its potential benefits in human health, as well as to assess the scientific evidence of traditional use and provide a basis for future research directions. Material and methods Peer-reviewed articles on Stephania rotunda were acquired via an electronic search of the major scientific databases (Pubmed, Google Scholar, and ScienceDirect). Data were collected from scientific journals, theses, and books. Results The traditional uses of Stephania rotunda were recorded in countries throughout Southeast Asia (Cambodia, Vietnam, Laos, and India). Different parts of Stephania rotunda were used in traditional medicine to treat about twenty health disorders. Phytochemical analyses identified forty alkaloids. The roots primarily contain l-tetrahydropalmatine (l-THP), whereas the tubers contain cepharanthine and xylopinine. Furthermore, the chemical composition differs from one region to another and according to the harvest period. The alkaloids exhibited approximately ten different pharmacological activities. The main pharmacological activities of Stephania rotunda alkaloids are antiplasmodial, anticancer, and immunomodulatory effects. Sinomenine, cepharanthine, and l-stepholidine are the most promising components and have been tested in humans. The pharmacokinetic parameters have been studied for seven compounds, including the three most promising compounds. The toxicity has been evaluated for liriodenine, roemerine, cycleanine, l-tetrahydropalmatine, and oxostephanine. Conclusion Stephania rotunda is traditionally used for the treatment of a wide range of ailments. Pharmacological investigations have validated different uses of Stephania rotunda in folk medicine. The present review highlights the three most promising compounds of Stephania rotunda , which could constitute potential leads in various medicinal fields, including malaria and cancer.

From the stem barks of Jacaranda mimosifolia benzoic acid (1), 1-naphthaleneacetic acid, 5-carboxy- 1,2,3,4,4a,7,8,8a-octahydro-1,2,4a-trimethyl-[1S-(1α,2β,4aβ,8aα)] ( 2), betulinic acid ( 3), lupeol (4) and ursolic acid (5) were isolated. Similarly, lapachol (6), dehydro-α-lapachone (7), 2- acetylfuro-1, 4-naphthoquinone (8), p-coumaric acid (9), caffeic acid (10), nonacosanoic acid, 2-(4-hydroxyphenyl)ethyl ester (11), β-sitosterol (12), kigelinol (13), oleanolic acid (14), β-friedelinol (15), pomolic acid (16), and kojic acid (17) were isolated from the stem barks of Kigelia africana. All the isolated compounds were characterized by using spectroscopic methods especially 1D and 2D NMR and ESI mass spectrometry and comparison with literature data. To the best of our knowledge, compounds 1, 2, 3 and 5, and compounds 11, 14, 15 and 16 were isolated for the first time from Jacaranda mimosifolia and Kigelia africana, respectively. All these compounds were screened for anticandidal activity by agar diffusion method and microbroth dilution technique on four Candida albicans strains (ATCCL26, ATCC12C, ATCCP37039, and ATCCP37037). Among them, compounds 9, 10, and 17 exhibited the highest anticandidal activity that varied between the microbial species (MIC= 0.01 ± 0.00 − 0.03 ± 0.00 mg/mL) on C. albicans ATCCL26, ATCCP37037, ATCCP37039 and ATCC12C strains. Compound 17 was likely the most active against the four Candida albicans strains (MIC= 0.01 ± 0.00 − 0.02 ± 0.00 mg/mL).

The tuber of Stephania rotunda is used to treat malaria in traditional medicine in many Asian countries. Cepharanthine is a bioactive bisbenzylisoquinoline alkaloid of the plant, in particular it presented antiplasmodial activity. Two modern and ecologic technologies were proposed to extract cepharanthine from S. rotunda tuber: ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE). Various parameters of these processes were studied to optimize the extraction of cepharanthine from the tuber. Quantification of cepharanthine was performed by a validated HPLC method. MAE in closed system, for 15 min, with the ratio solid–liquid 1/30, the solvent ethanol–water (50:50, v/v), at 80 °C and 100 W, was the most effective conditions on the extraction yield. UAE was the most ecologic because it did not need any heating and was shorter than MAE (10 min). Statistical analyses showed that these two methods exhibited the same efficiency for the extraction of cepharanthine and both methods were more efficient than percolation. MAE and UAE permitted the best extraction of cepharanthine from the S. rotunda tuber when the solvent used was ethanol–water in comparison with dichloromethane. Both eco-extractions, using ethanol–water, were found to be promising alternative methods for the extraction of cepharanthine from S. rotunda.

International audience; Contact irritant and non-contact repellent activities of β-caryophyllene oxide were evaluated against laboratory strains of female Aedes aegypti (USDA strain), a major arbovirus vector and Anopheles minimus (KU strain), a major malaria parasite vector, compared with the synthetic repellent DEET, using an excito-repellency test system. β-caryophyllene oxide and DEET were tested at concentrations of 0.1, 0.25, 0.5 and 1.0% (v/v). Anopheles minimus was found to be more sensitive to β-caryophyllene oxide than that of Ae. aegypti and exhibited high avoidance response rates (86–96% escape) at 0.5% and 1.0% concentrations in contact and non-contact trials compared with Ae. aegypti (22–59% escape). However, at the same concentrations, DEET displayed lower irritancy and repellency capacities against these two mosquito species (range 0–54% escape) compared to β-caryophyllene oxide. The analysis of escape responses showed significant differences between mosquito species at all concentrations (P

Malaria remains a major public health problem due to the emergence and spread of Plasmodium falciparum drug resistance. There is an urgent need to investigate new sources of antimalarial drugs which are more effective against Plasmodium falciparum. One of the potential sources of antimalarial drugs is traditional medicinal plants. In this work, we studied the in vitro antiplasmodial activity of chloromethylenic, methanolic, and MeOH/H2O (1/1) crude extracts and decoction obtained from eight medicinal plants collected in Burkina Faso and of total alkaloids for five plants. Extracts were evaluated in vitro for efficacy against Plasmodium falciparum strain K1, which is resistant to chloroquine, pyrimethamine and proguanil using the fluorescence-based SYBR Green I assay. The antiproliferative activity on human-derived hepatoma cell line HepG2 and Chinese hamster ovary (CHO) cells was evaluated using the 3-[4,5-dimethylthyazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) test in order to determine the selectivity index. Among the plant extracts tested for in vitro antiplasmodial activity, 16 were considered to be inactive (with IC50 > 10 μg/ml), six showed a moderate activity (5 < IC50 ≤ 10 μg/ml), and six were found to have a good in vitro activity with IC50 value ≤ 5 μg/ml. The highest antiplasmodial activity was found for extracts from: the alkaloid leaf extract and the chloromethylenic extracts of Combretum fragrans (IC50 = 3 μg/ml, IC50 = 5 μg/ml), the total alkaloids and the chloromethylenic leaf extracts of Combretum collinum (IC50 = 4 μg/ml), the MeOH/H2O leaf extract of Terminalia avicennioides (IC50 = 3.5 μg/ml), and the alkaloid leaf extract of Pavetta crassipes (IC50 = 5 μg/ml). Three other extracts showed moderate antiplasmodial activity (5 < IC50 ≤ 10 μg/ml): Terminalia avicennioides and Combretum fragrans methanolic extracts and Acacia kirkii alkaloid leaf extract (IC50 = 6.5, 9 and 10 μg/ml respectively). The Terminalia avicennioides crude MeOH/H2O (80:20 v/v) extract of the leaves was submitted to a successive liquid/liquid extraction with ethylacetate and n-butanol respectively. The extracts were investigated for in vitro antiplasmodial activity and antioxidant properties using DPPH(·), ABTS(+) and FRAP methods. The ethylacetate extract showed the best antiplasmodial activity (7 μg/ml) and the active constituent was isolated as ellagic acid by bioguided fractionation with an IC50 = 0.2 μM on Plasmodium falciparum and SI = 152. Besides, Terminalia avicennioides leaf extract and ellagic acid showed a good antioxidant activity. Our finding confirms the importance of investigating the antimalarial activity of plant species used in traditional medicine. Overall, two plants belonging to the Combretaceae family, Combretum fragrans and Combretum collinum appeared to be the best candidates and will be further investigated for their antiplasmodial properties, in order to isolate the molecules responsible for the antiplasmodial activity.

Strychnos nux-vomica L. (Loganiaceae) is famous for its monomeric alkaloid content, such as strychnine, a convulsant poison. The stem bark of the tree is traditionally used to treat intermittent fever in South East Asia. In various studies, it appeared that dimeric indolo-monoterpenic alkaloids possess a promising activity on Plasmodium falciparum. Three bisindolomonoterpenic alkaloids together with strychnochrysine, previously identified in the root bark of S. nux-vomica, were isolated from the stem bark. The structures of these compounds were established using NMR spectroscopy and mass spectrometry. Stereochemistry of the compounds was confirmed by molecular modelling. This then allowed the structural determination of strychnoflavine, a coloured bisindole alkaloid previously isolated from the root bark of the tree. Moreover, the conformational inversion in alkaloids possessing an ether bond in the strychnane moiety could be easily predicted by specific δ (13)C NMR values. These longicaudatine-type alkaloids were found to display in vitro antiplasmodial activity against a chloroquine resistant strain and a chloroquine sensitive strain. The most interesting was strychnochrysine showing an IC(50) value at around 10 μM.

Ethnopharmacological relevance Stephania rotunda Lour. ( Menispermaceae ) is a creeper growing in many countries of Asia and commonly found in the mountainous areas of Cambodia. As a folk medicine, it has been mainly used for the treatment of fever and malaria. The pharmacological activity is mostly due to alkaloids. Thus the aim of this study is to isolate new bioactive alkaloids from Stephania rotunda and to evaluate their in vitro antiplasmodial activity. Materials and methods Alkaloids were isolated and identified from dichloromethane and aqueous extracts using a combination of flash chromatography, high performance liquid chromatography, mass spectrometry and nuclear magnetic resonance. The purified compounds were tested for in vitro antiplasmodial activity on chloroquine-resistant W2 strain of Plasmodium falciparum . Results A new aporphine alkaloid named vireakine ( 2 ) along with two known alkaloids stephanine ( 1 ) and pseudopalmatine ( 8 ), described for the first time in Stephania rotunda , and together five known alkaloids tetrahydropalmatine ( 3 ), xylopinine ( 4 ), roemerine ( 5 ) , cepharanthine ( 6 ) and palmatine ( 7 ) were isolated and identified. The structure of the new alkaloid was established on the basis of 1D and 2D NMR experiments and mass spectrometry. The compounds were evaluated for their in vitro antiplasmodial and cytotoxic activities. All tested compounds showed significant antiplasmodial activities with IC 50 ranged from 1.2 μM to 52.3 μM with a good selectivity index for pseudopalmatine with IC 50 of 2.8 μM against W2 strain of Plasmodium falciparum and IC 50 >25 μM on K562S cells. Conclusions This study provides evidence to support the use of Stephania rotunda for the treatment of malaria and/or fever by the healers. Alkaloids of the tuber exhibited antiplasmodial activity and particularly cepharanthine and pseudopalmatine.

International audience; A study of the behavioral responses of Aedes aegypti and Anopheles minimus to 3 Cambodian plant extracts at 3 different concentrations (1%, 2.5%, and 5%) was performed using an excito-repellency test system. These 3 plants were Strophanthus scandens, Capparis micracantha, and Dioscorea hispida, selected according to traditional healer's knowledge, bibliographic studies and market surveys. Results showed that S. scandens leaves' hexane extract was the only one to exert repellency against Ae. aegypti with 23.3% of escaped mosquitoes at a concentration of 5%. Capparis micracantha was responsible for an irritant activity against An. minimus with 20.2% of escaped mosquitoes at a concentration of 2.5% and 22.8% escaping at a concentration of 5%. Dioscorea hispida showed an irritant activity on both mosquito species with 23.2% of escaped Ae. aegypti at a concentration of 5% and about 20% of escaped An. minimus at 2.5% and 5%. This is the first report on the irritant and repellent activities of S. scandens , D. hispida , and C. micracantha against mosquito species.

This research paper presents a quick and ecofriendly technique for the extraction of harpagoside (HS), the active marker of Harpagophytum procumbens (HP), along with a comparison with conventional methods so as to propose an efficient HPLC method. HP is widely used as an anti-inflammatory in phytotherapy. The quality control of the herbal drug and extract calls for a time consuming method of conventional extraction, which involves a high consumption of solvents. In this study, HP has been extracted using conventional ultrasound (UAE) and microwave (MAE)-assisted methods. The effects have been examined based on several parameters of HS extraction efficiencies. An HPLC method with a core-shell column was developed in order to calculate the HS in HP. The flow rate was reduced by 4. The method of validation used is specific, linear, precise and accurate. MAE and UAE saved solvent consumption, time and energy. It has, therefore, been found that the combined UAE-HPLC process is convenient and appropriate for the quality control of HP.

International audience

International audience

Stephania rotunda (Menispermaceae), a creeper commonly found in the mountainous areas of Cambodia, has been mainly used for the treatment of fever and malaria. Thus, the aim of this study is to investigate the chemical composition and antiplasmodial activity of different samples of S. rotunda and compare their antiplasmodial activity with their alkaloid content. Sixteen samples from different parts (roots, stem, and tuber) of S. rotunda were collected from four regions of Cambodia (Battambang, Pailin, Siem Reap, and Kampot). Reversed-phase HPLC was used to determine the content of three bioactive alkaloids (cepharanthine, tetrahydropalmatine, and xylopinine). These three alkaloids have been found in all samples from Battambang and Pailin (samples I–IX), whereas only tetrahydropalmatine was present in samples from Siem Reap and Kampot (samples X–XVI). The analyzed extracts were evaluated for their antiplasmodial activity on W2 strain of Plasmodium falciparum. Among them, 13 extracts were significantly active with inhibitory concentration 50 (IC50) from 1.2 to 3.7 µg/mL and 2 extracts were moderately active (IC50 = 6.1 and 10 µg/mL, respectively), whereas sample XI was not active (IC50 = 19.6 µg/mL). A comparison between antiplasmodial activity and concentration of the three bioactive alkaloids in S. rotunda extracts has been realized. Copyright © 2012 John Wiley & Sons, Ltd.

Ethnopharmacological relevance Bark extracts of Nauclea latifolia, Nauclea diderrichii, Nauclea pobeguinii and Nauclea vandergutchii are used in traditional medicine in West and South Africa for the treatment of fevers, diarrhea and malaria. Aim of the study To estimate the possible long-term toxicity and genotoxicity of plant extracts (dichloromethane, methanol, water/methanol, water) and saponins. Materials and methods The clastogenicity of plant extracts and saponins was assessed by the micronucleus assay performed on Chinese Hamster Ovary cells. The DNA-damaging activity of saponin mixture was assessed by the comet assay on Chinese Hamster ovary cells. Results Hydromethanolic extracts from Nauclea latifolia, Nauclea diderrichii and Nauclea pobeguinii exhibited a significant clastogenic/aneugenic activity without S9 mix. The hydromethanolic extract from Nauclea diderrichii was the most clastogenic/aneugenic fraction with a Minimal Active Concentration (MAC) of 23.1 μg mL−1. It was submitted to a separation step leading to six main saponins identified as quinovic acid glycosides (saponins A, D, E, G, J, K). None of the isolated saponins exerted a significant clastogenic/aneugenic activity by the micronucleus assay, however a mixture made with equal quantities of each of the six saponins exhibited a direct genotoxic/clastogenic activity as assessed by both the micronucleus assay and the comet assay on Chinese Hamster Ovary cells. Conclusion Saponins present in the hydromethanolic extracts of Nauclea induced synergistic in vitro DNA-damage and chromosome mutations in mammalian cells. This genotoxic activity was probably due to the capacity of Nauclea saponins to reduce cell defense against oxidative stress through the inhibition of glutathione-S-transferase activity.

Three bisbenzylisoquinoline alkaloids were isolated for the first time from Stephania rotunda tuber. Their structures were elucidated by spectroscopic methods and their antiplasmodial activity was investigated in vitro on chloroquine resistant Plasmodium falciparum strain W2. These alkaloids were identified as 2-norcepharanthine (1), cepharanoline (2) and fangchinoline (3). In vitro, they displayed significant antiplasmodial activity with inhibitory concentration 50 values of 0.3, 0.2 and 0.3 µM.

Three new triterpene saponins, leonticins I (1), J (2) and L (3) were isolated from the tubers of Leontice smirnowii. On the basis of spectroscopic methods, including 2D NMR experiments (DEPT, gs-COSY, gs-HMQC, gs-HMBC and gs-HSQC-TOCSY), mass spectrometry (HR-ESI-MS) and chemical degradation, the structures of the new compounds were elucidated as 3-O-β-D-glucopyranosyl-(1 → 3)-[β-D-xylopyranosyl-1 → 2)]-α-L-arabinopyranosyl-28-O-[α-L-rhamnopyranosyl-(1 → 4)-β-D-glucopyranosyl-(1 → 6)-β-D-glucopyranosyl]-3β-hydroxy-30-norolean-12,20(29)-dien-28-oic acid (1), 3-O-[β-D-xylopyranosyl-(1 → 3)-β-D-galactopyranosyl-(1 → 4)-β-D-glucopyranosyl-(1 → 3)-α-L-arabinopyranosyl]-28-O-[α-L-rhamnopyranosyl-(1 → 4)-β-D-glucopyranosyl-(1 → 6)-β-D-glucopyranosyl]-3β-hydroxy-30-norolean-12,20(29)-dien-28-oic acid (2) and 3-O-[β-D-xylopyranosyl-(1 → 3)-β-D-galactopyranosyl-(1 → 4)-β-D-glucopyranosyl-(1 → 3)]-[β-D-xylopyranosyl-(1 → 2)]-α-L-arabinopyranosyl]-28-O-[α-L-rhamnopyranosyl-(1 → 4)-β-D-glucopyranosyl-(1 → 6)-β-D-glucopyranosyl]-3β-hydroxy-30-norolean-12,20(29)-dien-28-oic acid (3), respectively. The aglycone 3β-hydroxy-30-norolean-12,20(29)-dien-28-oic acid was observed for the first time in Leontice species.

Together with apigenine dimethylether and piperchabamide A, a new amide alkaloid, Kaousine and the Z form of antiepilepsirine were isolated from the aerial part of Piper capense L.f (Piperaceae). Their structures were elucidated by spectrometric methods and their in vitro antiparasitic activities were evaluated on Plasmodium falciparum.

Gentiopicroside (1) is the major secoiridoid glucoside constituent of Cephalaria kotschyi roots. The mutagenicity, DNA-damaging capacities, and clastogenicity of this molecule were evaluated by the Salmonella typhimurium mutagenicity assay (Ames test) on tester strains TA97a, TA98, TA100, and TA102, the alkaline comet assay, and the micronucleus assay on CHO cells. All tests were performed with and without the metabolization mixture, S9 mix. In the Ames test, the mutagenicity of 1 was limited to TA102 without S9 mix (2.3 rev microg(-1)). The genotoxicity was more evident without S9 mix (0.78 OTMchi(2) units microg(-1) mL) than with the metabolic mixture (0.16 OTMchi(2) units microg(-1) mL) with the comet assay. Similarly, the clastogenicity without S9 mix was 0.99 MNC microg(-1) mL and 0.38 MNC microg(-1) mL with S9 mix in the micronucleus assay. The interaction of 1 with DNA is probably through the involvement of oxidative DNA lesions.

In order to prevent the destruction of the ecology and to sustain the flora mainly for medicinal plants, we investigated on alternative parts taken from four plants already known to display antiplasmodial activities and largely used by traditional healers in sub-Saharan Africa. The evaluated parts are bark of trunk for Zanthoxylum zanthoxyloides and leaves for Sarcocephalus latifolius instead of roots, and leaves for Combretum molle and Anogeissus leiocarpus instead of stem bark. The antiplasmodial activity of extracts of these plants was evaluated in vitro using the multi-resistant strain (W2) of Plasmodium falciparum. Antiproliferative activity was also assessed, using K562S human monocyte cell lines, along with calculation of the selectivity index (SI) of each extract. The highest in vitro antiplasmodial activity was found in the alkaloid extract of trunk bark from Z. zanthoxyloides and from the MeOH extract of A. leiocarpus leaves (IC(50) = 1.2 microg/mL and 4.9 microg/mL, respectively) with good selectivity index. Moderate activity was found in the MeOH extract (IC(50) = 5.7 microg/mL) and MeOH/H2O extract (IC(50) = 7.9 microg/mL) of C. molle leaves. Moderate activity was also found in the MeOH/H20 extract (IC(50) = 5.2 microg/mL) and the decoction (IC(50) = 8.2 microg/mL) from leaves of A. leiocarpus. No good activity was found with extracts from roots of S. latifolius. All extracts tested displayed low levels of cytotoxicity against K562S cells. The data generated clearly show that the trunk bark for Z. zanthoxyloides and the leaves for A. leiocarpus and C. molle could be used for the treatment of malaria instead of roots and stem bark.

Three major alkaloids: cepharanthine (1), tetrahydropalmatine (2) and xylopinine (3) isolated from Stephania rotunda tuber were investigated for their cytotoxic activity in a panel of human cancer cells (HT29, LS174T, SW620 and HepG2) using MTT assay. In the present study, cepharanthine (1) exerted potent cytotoxicity against colon and hepatoma cancer cell lines with IC50 values between 2.4 and 5.3 µM while tetrahydropalmatine (2) and xylopinine (3) displayed weak cytotoxicity. In addition, the mutagenic activity of cepharanthine (1) was investigated using a modified liquid incubation technique of the Salmonella/microsomal assay. This alkaloid (1) was found to be non-mutagenic for doses up to 8.2 µM. Copyright © 2008 John Wiley & Sons, Ltd.

The aim of this study was to investigate the ability of α-hederin to improve the efficacy of widely prescribed 5-fluorouracil (5-FU) in a human colon adenocarcinoma model. Drug combinations of α-hederin and 5-FU using both fixed-concentration and combination index methods were performed in vitro in HT-29 cells. The results showed that α-hederin at sub-IC50 cytotoxic concentrations enhanced 5-FU cytotoxicity about 3.3-fold (p < 0.001). Simultaneous combination of α-hederin and 5-FU at their IC50 ratio showed either a synergistic effect at a moderate cytotoxic range (25% of cell growth inhibition) or an antagonistic effect at a high level of growth inhibition. The data indicate therefore that it is possible to optimize colorectal cancer cell sensitivity to 5-FU with α-hederin. Copyright © 2008 John Wiley & Sons, Ltd.

Three new oleanane-type saponins, giganteosides L (1), M (2) and N (3) along with eight known ones were isolated from the roots of Cephalaria gigantea. Their structures were established as 3-O-[beta-D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl]-28-O-[beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl]-oleanolic acid, 3-O-[beta-D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl]-28-O-[beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl]-hederagenin, 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucuronopyranosyl]-28-O-[beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl]-hederagenin, respectively, by means of spectroscopic methods (1D and 2D NMR, HR-ESI-MS). Cytotoxic activity of monodesmosides was investigated in vitro using three cancer cell lines, namely, human non pigmented melanoma MEL-5 and human leukemia HL-60. Giganteosides D (4) and E (5) showed antiproliferative effect on human cell lines with IC(50) values in the range 3.15-7.5 microM.

International audience; DIG, a liquid herbal preparation made from a mixture of diluted mother tinctures of Berberis vulgaris, Taraxacum officinale and Arctium lappa, was assessed for its antimutagenic properties against mitomycin C. The micronucleus assay on Chinese hamster ovary (CHO)-K1 cells was used to evaluate the in vitro anticlastogenic activity of DIG compared to those of separately diluted mother tinctures. The micronucleus assay was performed on mouse erythrocytes and the comet assay was performed on mouse liver, kidney, lung, brain and testicles to assess the protective effects of DIG (0.2 and 2 % at libitum) against an intraperitoneal injection of mitomycin C (1 mg Kg(-1)) in mice. DIG exerted a powerful anticlastogenic activity, under both pretreatment and simultaneous treatment conditions as assessed by the micronucleus assay in CHO-K1 cells. Its protective activity was greater than that observed for each mother tincture. DIG reduced micronuclei levels in mouse erythrocytes and suppressed \textgreater 80 % of DNA strand breaks in the liver, kidney, lung, brain and testicles of mice exposed to mitomycin C.