Patrick Valere Tsouh Fokou

Ethnopharmacological survey of Annonaceae medicinal plants used to treat malaria in four areas of Cameroon

ETHNOPHARMACOLOGICAL RELEVANCE Malaria endemic countries have vital resources that are medicinal plants on which their traditional medicines depend. In some Cameroonian settings, in addition to the commonly used potions from plants like Alstonia boonei, Zanthoxylum macrophylla and Mangifera indica, other herbal species are being increasingly used to treat malaria. So, specialized traditional healers have developed alternative reasonably priced therapies, relying on the signs and/or symptoms of malaria. Within this framework, Annonaceae plants were found to be increasingly utilized and therefore, highlighting the need to document this traditional knowledge for better malaria control. MATERIALS AND METHODS Interview approach was used to document indigenous knowledge, usage customs and practices of Annonaceae species in the treatment of malaria in four Cameroonian areas (Yaoundé and its surroundings, Ngoyang, Kon-Yambetta and Mbalmayo). RESULTS A total of 19/30 users of plants accepted to share their experiences during a semi-structured survey. Twelve of the respondents were men and seven were women. Thirty recipes based on twenty-one plants were recorded. CONCLUSION Annickia chlorantha was the only plant commonly found in the four study sites. Seven species of Annonaceae were found to be used to treat malaria, while 14 were used to treat symptoms that might be related to malaria.

Repurposing the Open Access Malaria Box To Discover Potent Inhibitors of Toxoplasma gondii and Entamoeba histolytica

ABSTRACT Toxoplasmosis and amebiasis are important public health concerns worldwide. The drugs currently available to control these diseases have proven limitations. Therefore, innovative approaches should be adopted to identify and develop new leads from novel scaffolds exhibiting novel modes of action. In this paper, we describe results from the screening of compounds in the Medicines for Malaria Venture (MMV) open access Malaria Box in a search for new anti-Toxoplasma and anti-Entamoeba agents. Standard in vitro phenotypic screening procedures were adopted to assess their biological activities. Seven anti-Toxoplasma compounds with a 50% inhibitory concentration (IC50) of <5 μM and selectivity indexes (SI) of >6 were identified. The most interesting compound was MMV007791, a piperazine acetamide, which has an IC50 of 0.19 μM and a selectivity index of >157. Also, we identified two compounds, MMV666600 and MMV006861, with modest activities against Entamoeba histolytica, with IC50s of 10.66 μM and 15.58 μM, respectively. The anti-Toxoplasma compounds identified in this study belong to scaffold types different from those of currently used drugs, underscoring their novelty and potential as starting points for the development of new antitoxoplasmosis drugs with novel modes of action.

Potent antiplasmodial extracts from Cameroonian Annonaceae

AIM OF THE STUDY In a search for new antimalarial leads, we have carried out a preliminary ethnopharmacological study with the aim of evaluating the in vitro antiplasmodial activity of extracts from thirteen Annonaceae species growing in Cameroon, and of assessing the acute toxicity of promising fractions in Swiss albino mice. MATERIALS AND METHODS Plants were selected on the basis of an ethnobotanical survey carried out in four sites in centre and south regions of Cameroon (Yaoundé neighbourhoods, Kon-Yambetta, Ngobayang and Mbalmayo) on Annonaceae plants locally used to treat malaria and related symptoms. The choice of the sites was mainly based on environmental factors enabling mosquito breeding, cosmopolitan areas regrouping people from different cultural origins, areas with limited access to health centers, and areas with people relying exclusively on traditional medical practices. Collected materials were extracted by maceration in 95% ethanol. The crude extract was partitioned using organic solvents and the fractions afforded were evaluated for antiplasmodial activity in culture against the W2 strain of Plasmodium falciparum. Promising fractions (methanol fractions) were assessed for their acute toxicity in Swiss albino mice. RESULTS From the results achieved, 37 (31.3%) out of 118 extracts tested exhibited antiplasmodial activity, with IC(50) values ranging from 1.07 μg/ml to 9.03 μg/ml. Of the active extracts, 29 (78.4%) were methanol fractions, 21 (72.4%) of which inhibited the parasites with IC(50)<5 μg/ml. The promising fractions proved to be safe through oral administration in mice. CONCLUSIONS The activities and toxicity profiles of methanol fractions indicate that they deserve to be further investigated in detail for antimalarial lead discovery.

Extracts from Annona Muricata L. and Annona Reticulata L. (Annonaceae) Potently and Selectively Inhibit Plasmodium Falciparum

The aim of this work was to screen extracts from Annona muricata and Annona reticulata in vitro against Plasmodium falciparum. Crude ethanolic extracts, methylene chloride fractions, aqueous fractions, subfractions and isolated compounds (stigmasterol-3-O-β-d-glucopyranoside, lichexanthone, gallic acid and β-sitosterol-3-O-β-d-glucopyranoside) were tested for cytotoxicity on erythrocytes and Human Foreskin Fibroblasts cells and against the W2 strain of P. falciparum in culture. Results indicated that none of the extracts was cytotoxic at concentrations up to 10 µg/mL. Most of the extracts, fractions and subfractions inhibited the growth of P. falciparum with IC50 values ranging from 0.07 to 3.46 µg/mL. The most potent was the subfraction 30 from A. muricata stem bark (IC50 = 0.07 µg/mL) with a selectivity index of ˃ 142. Subfraction 3 from A. muricata root also exhibited very good activity (IC50 = 0.09 µg/mL) with a high selectivity index (SI ˃ 111). Amongst the isolated compounds, only gallic acid showed activity with IC50 of 3.32 µg/mL and SI > 10. These results support traditional claims for A. muricata and A. reticulata in the treatment of malaria. Given their limited cytotoxicity profile, their extracts qualify as promising starting points for antimalarial drug discovery.

Update on Medicinal Plants with Potency on Mycobacterium ulcerans.

Mycobacterium ulcerans disease has been a serious threat for people living in rural remote areas. Due to poverty or availability of traditional medicine these populations rely on herbal remedies. Currently, data on the anti-Mycobacterium ulcerans activity of plants, so far considered community-based knowledge, have been scientifically confirmed, concomitantly with some medicinal plants used to treat infectious diseases in general. Products derived from plants usually responsible for the biological properties may potentially control Mycobacterium ulcerans disease; numerous studies have aimed to describe the chemical composition of these plant antimicrobials. Thus, the present work provides the first compilation of medicinal plants that demonstrated inhibitory potential on Mycobacterium ulcerans. This work shows that the natural products represent potential alternatives to standard therapies for use as curative medicine for Mycobacterium ulcerans disease.

Marine Algae as Source of Novel Antileishmanial Drugs: A Review

Leishmaniasis is a vector-borne neglected tropical disease caused by protozoan parasites of the Leishmania genus and transmitted by the female Phlebotomus and Lutzomyia sand flies. The currently prescribed therapies still rely on pentavalent antimonials, pentamidine, paromomycin, liposomal amphotericin B, and miltefosine. However, their low efficacy, long-course treatment regimen, high toxicity, adverse side effects, induction of parasite resistance and high cost require the need for better drugs given that antileishmanial vaccines may not be available in the near future. Although most drugs are still derived from terrestrial sources, the interest in marine organisms as a potential source of promising novel bioactive natural agents has increased in recent years. About 28,000 compounds of marine origin have been isolated with hundreds of new chemical entities. Recent trends in drug research from natural resources indicated the high interest of aquatic eukaryotic photosynthetic organisms, marine algae in the search for new chemical entities given their broad spectrum and high bioactivities including antileishmanial potential. This current review describes prepared extracts and compounds from marine macroalgae along with their antileishmanial activity and provides prospective insights for antileishmanial drug discovery.

In Vitro Activity of Selected West African Medicinal Plants against Mycobacterium ulcerans Disease

Buruli ulcer (BU) is the third most prevalent mycobacteriosis, after tuberculosis and leprosy. The currently recommended combination of rifampicin-streptomycin suffers from side effects and poor compliance, which leads to reliance on local herbal remedies. The objective of this study was to investigate the antimycobacterial properties and toxicity of selected medicinal plants. Sixty-five extracts from 27 plant species were screened against Mycobacterium ulcerans and Mycobacterium smegmatis, using the Resazurin Microtiter Assay (REMA). The cytotoxicity of promising extracts was assayed on normal Chang liver cells by an MTT assay. Twenty five extracts showed activity with minimal inhibitory concentration (MIC) values ranging from 16 µg/mL to 250 µg/mL against M. smegmatis, while 17 showed activity against M. ulcerans with MIC values ranging from 125 µg/mL to 250 µg/mL. In most of the cases, plant extracts with antimycobacterial activity showed no cytotoxicity on normal human liver cells. Exception were Carica papaya, Cleistopholis patens, and Polyalthia suaveolens with 50% cell cytotoxic concentrations (CC50) ranging from 3.8 to 223 µg/mL. These preliminary results support the use of some West African plants in the treatment of Buruli ulcer. Meanwhile, further studies are required to isolate and characterize the active ingredients in the extracts.

In vitro antimycobacterial activity of six Cameroonian medicinal plants using microplate alamarBlue assay.

OBJECTIVE/BACKGROUND The latest incidence of tuberculosis (TB) (per 100,000 people) in Cameroon was 243.00 as of 2011. Over the past 21 years, the value for this indicator has fluctuated between 112.00 in 1990 and 320.00 in 2003. Worldwide, this incidence has also increased, bringing back TB as a reemerging disease. On the same note, resistance to anti-TB drugs has increased, urging the search for new molecules. METHODS This study was carried out to evaluate the antimycobacterial activity of six medicinal plants on the virulent strain, H37Rv, using the microplate alamarBlue assay. Mycobacterium tuberculosis (H37Rv strain) was incubated with decreased concentrations of six plant extracts, ranging from 250 μg/mL to 31.25 μg/mL. After 7 days of incubation at 37 °C, the effects of these plant extracts on the viability of the mycobacteria were evaluated. For each plant extract, the minimal inhibitory concentration was determined. RESULTS The results showed that the compounds MBC1, MBC24, MBC68, MBC81, MBC117, and MBC118 were the best candidates with minimal inhibitory concentrations of 31.25, 62.5, 125, 62.5, and 125 μg/mL, respectively. CONCLUSION These results confirm and validate the traditional use of these plants to treat respiratory diseases, which could be good sources and alternatives of plant metabolites for anti-TB-drug development.

Antimycobacterial potency and cytotoxicity study of three medicinal plants

Objective/Background: Mycobacterial infections including tuberculosis, leprosy, and buruli ulcer are among the most prevalent, debilitating, and deadly tropical diseases, especially in Sub-Saharan Africa. The development of drug resistance to the currently available drugs and the poor compliance emphasize the need for new chemotherapeutic agents. This study was designed to evaluate the in vitro activity of Cleistopholis patens, Annona reticulata, and Greenwayodendron suaveolens against Mycobacterium smegmatis. The safety on normal liver cells was also assessed. Methods: The crude extracts, fractions, and subfractions were tested against M. smegmatis and for cell cytotoxicity on WRL-68, normal human hepatocyte using microdilution resazurin-based assays. The phytochemical screening was performed using standard methods. Results: Most of the extracts, fractions, and subfractions inhibited the growth of M. smegmatis with minimum inhibitory concentration (MIC) values ranging from 6.25 μg/mL to 125 μg/mL. The subfractions P12 and P29 from G. suaveolens twig were more potent with MIC values of 6.25 μg/mL and 25 μg/mL, respectively. Fruit crude extract and root CH2Cl2 fraction from A. reticulata also showed activity with MIC values of 50 μg/mL and 25 μg/mL, respectively. Crude extracts from the twig and stem bark of C. patens displayed inhibition at MIC values of 125 μg/mL and 100 μg/mL, respectively. Majority of active extracts showed no cell cytotoxicity, except the extract from C. patens with IC50 ranging from 41.40 μg/mL to 93.78 μg/mL. The chemical investigation of the promising extracts revealed the presence of phenols, alkaloids, glycosides, triterpenes, and acetogenins. Conclusion: The results achieved from this preliminary antimycobacterial drug discovery study supported the traditional claims of C. patens, A. reticulata, and G. suaveolens in the treatment of mycobacterial infections. Meanwhile, further fractionation is required to characterize the active ingredients.

Antimycobacterial ingredients from plants used in traditional medicine to treat Buruli ulcer.

Aim and objectives: Buruli ulcer (BU) is a neglected tropical disease caused by a mycobacteria, Mycobacterium ulcerans. The WHO recommended Rifampicin-Streptomycin combination side effects and poor compliance, leaves rural populations with no choice than to patronise indigenous remedies. This study is aimed at validating medicinal plants used in traditional medicine to treat BU by investigating the in vitro efficacy and safety as well as their composition in active molecules. Methods: A short report-based survey was used to identify medicinal plants used traditionally for BU treatment. Maceration of collected plant samples in methanol, hydroethanolic, ethanol, dichloromethane, and hexane, resulted in a total of 67 extracts assessed for antimycobacteria activity against Mycobacterium smegmatis and Mycobacterium ulcerans using the Resazurin Microtiter Assay. The cytotoxicity effect of promising extracts was assessed on normal human liver cells using the MTT assay. The bio-guided fractionation of the promising extracts led to the isolation of active compounds. Results: Majority of plants prepared as infusion, decoction, poultice, and macerate were administered topically. Significant antimycobacterial activity with MIC values ranging from 16 to 250 μg/mL was recorded against M. smegmatis (25 extracts) and M. ulcerans (17 extracts).1 Most of antimycobacterial extracts showed no significant cytotoxicity against normal human hepatocytes. 1The isolation guided by the biological activity revealed nine compounds with significant in vitro anti-M. ulcerans activity (MIC = 16–128 μg/mL). Conclusions: The results completed support the use these plants in the indigenous knowledge against BU. Further analyses of active principles might lead to new drug toe fight against BU.