Evaluation of macrofilaricidal and microfilaricidal activities against Onchocerca ochengi and cytotoxicity of some synthesized azo compounds containing thiophene backbone.

Control and treatment of onchocerciasis, a devastating tropical filarial disease caused by Onchocerca volvulus, rely solely on the community directed treatment with ivermectin. However, ivermectin is only microfilaricidal with evidence of resistance of the parasite among other limitations, which necessitate the search for new efficacious and safe filaricides. Ten synthetic thienylazoryl dyes were screened in vitro against adult and microfilariae worm stages of Onchocerca ochengi based on worm motility and MTT formazan assay. Cytotoxicity of active compounds was assessed on monkey kidney epithelial cells (LLC-MK2) using the MTT formazan assay. Seven (7) compounds showed both macrofilaricidal activity against adult male worms and microfilaricidal activity among which three 4a, 4c and 4e recorded the highest activity (IC50 = 4.2 to 8.8µM) against adult male worms, comparable to some standard anthelmintics. Five compounds showed rapid activity against microfilariae with 100% inhibition after 24-h incubation. The active compounds were nontoxic on monkey kidney cells (CC50> 4µg/mL), but their selectivity index values were relatively low (≤ 3). The thienylazoaryls with both macrofilaricidal and microfilaricidal activities may yield molecules which could be used for eradication of onchocerciasis following further medicinal chemistry modification of their structures to enhance their selectivity.

In vitro antifungal, antibacterial activities and nutritional value of nine Cameroonian edible vegetables.

BACKGROUND: The high medicinal potential of plants, including edible vegetables, is well documented. Vegetables can afford significant health benefits to consumers, depending on their medicinal properties and nutritional value. This study analysed the antimicrobial activity and nutrient contents of nine commonly consumed Cameroonian vegetables and spices for which such information is at present limited. METHODS: The antimicrobial activity of the methanol extracts of the vegetables was evaluated by disc diffusion and microdilution methods against three tomato fungi and two pathogenic bacteria species. RESULTS: The inhibition zones against fungi ranged from 10–21 mm; Irvingia gabonensis and Apium graveolens showed the highest zones with dose-dependent activity against Fusarium solani and F. oxysporum. The inhibition zones against bacteria ranged from 8–12 mm with Allium porrum having the highest inhibition zone (12 mm). Irvingia gabonensis seeds had the lowest minimum inhibitory concentration (MIC) of 6.25 mg/mL against F. solani and also had the lowest MIC of 2 mg/mL against S. aureus. Proximate composition and mineral analysis were carried out on the most active antimicrobial vegetables, I. gabonensis seeds and A. graveolens. I. gabonensis seeds were rich in lipids (69.90 ±0.14%) while A. graveolens leaves were rich in protein (35.35 ±0.49%). For macro minerals, phosphorous had the highest concentration in I. gabonensis seeds (359.67 ±1.89 mg/100 g) and A. graveolens leaves (622.14 ±2.69 mg/100 g). Iron content was the highest of micro minerals in I. gabonensis seeds (276.51 ±1.41 mg/100 g) while zinc concentration was   the highest in A. graveolens leaves (16.86 ±0.27 mg/100 g). CONCLUSIONS: This study has shown that three of the nine Cameroonian vegetables, I. gabonensis seeds, Apium graveolens and A. porrum, may potentially offer both antimicrobial and nutritional benefits to consumers. Consequently, further studies should be conducted to ascertain the effect of cooking and other factors in order to maximize these benefits.

Structurally simple synthetic 1, 4-disubstituted piperidines with high selectivity for resistant Plasmodium falciparum.

BACKGROUND: Emergence of resistance to artemisinins and some of their combinations in chemotherapy of clinical malaria has intensified the search for novel safe efficacious antimalarial molecules. Fourteen synthetic 1, 4-disubstituted piperidines with simple molecular structures were evaluated in this study. METHODS: Antiplasmodial activity were determined against cultured chloroquine-sensitive 3D7 and resistant Dd2 strains of P. falciparum by in vitro parasite growth inhibition. A primary screen was done to identify active compounds by fluorescence microscopy followed by a secondary screen to determine IC50 and IC90 values of active compounds by the parasite lactate dehydrogenase assay. Cytotoxicity of active compounds was assessed using the MTT/formazan assay and selectivity indices (SIs) determined. Optical densities were analysed to obtain experimental results. RESULTS: The compounds produced 56 to 93% inhibition of parasite growth at 40 µg/mL. Eight compounds (2 ketone, 5 alcohol and one amine analogues) showed high activity (IC50s between 1 and 5 µg/mL). Nine compounds were highly selective for the parasite (SIs = 15 to 182). Three promising (alcohol) analogues were identified: [1-(4-fluorobenzyl) piperidin-4-yl] [4-fluorophenyl] methanol, (7), [1-(3, 4-dichlorobenzyl) piperidin-4-yl] [4- fluorophenyl] methanol (8) and [1-(4-bromobenzyl) piperidin-4-yl] [4- fluorophenyl] methanol (11) which were more active on the resistant strain (IC50 values between 1.03 to 2.52 µg/mL), than the sensitive strain (IC50 values between 2.51 to 4.43 µg/mL). CONCLUSIONS: The alcohol analogues were the most active and most selective for the parasite with three promising hit molecules identified among them, suggesting the hydroxyl group at C-7' in these alcohol analogues is contributing greatly to their antiplasmodial activity. Further exploration of the core structure using chemistry approaches and biological screening including in vivo studies in an animal model of malaria may yield important antimalarial leads.

4-aroylpiperidines and 4-(α-hydroxyphenyl)piperidines as selective sigma-1 receptor ligands: synthesis, preliminary pharmacological evaluation and computational studies.

BACKGROUND: Sigma (σ) receptors are membrane-bound proteins characterised by an unusual promiscuous ability to bind a wide variety of drugs and their high affinity for typical neuroleptic drugs, such as haloperidol, and their potential as alternative targets for antipsychotic agents. Sigma receptors display diverse biological activities and represent potential fruitful targets for therapeutic development in combating many human diseases. Therefore, they present an interesting avenue for further exploration. It was our goal to evaluate the potential of ring opened spipethiane (1) analogues as functional ligands (agonists) for σ receptors by chemical modification. RESULTS: Chemical modification of the core structure of the lead compound, (1), by replacement of the sulphur atom with a carbonyl group, hydroxyl group and 3-bromobenzylamine with the simultaneous presence of 4-fluorobenzoyl replacing the spirofusion afforded novel potent sigma-1 receptor ligands 7a-f, 8a-f and 9d-e. The sigma-1 receptor affinities of 7e, 8a and 8f were slightly lower than that of 1 and their selectivities for this receptor two to threefold greater than that of 1. CONCLUSIONS: It was found that these compounds have higher selectivities for sigma-1 receptors compared to 1. Quantitatitive structure-activity relationship studies revealed that sigma-1 binding is driven by hydrophobic interactions.Graphical abstractIdentified pharmacophore features for sigma binding.

New antimalarial hits from Dacryodes edulis (Burseraceae)--part I: isolation, in vitro activity, in silico "drug-likeness" and pharmacokinetic profiles.

The aims of the present study were to identify the compounds responsible for the anti-malarial activity of Dacryoedes edulis (Burseraceae) and to investigate their suitability as leads for the treatment of drug resistant malaria. Five compounds were isolated from ethyl acetate and hexane extracts of D. edulis stem bark and tested against 3D7 (chloroquine-susceptible) and Dd2 (multidrug-resistant) strains of Plasmodium falciparum, using the parasite lactate dehydrogenase method. Cytotoxicity studies were carried out on LLC-MK2 monkey kidney epithelial cell-line. In silico analysis was conducted by calculating molecular descriptors using the MOE software running on a Linux workstation. The "drug-likeness" of the isolated compounds was assessed using Lipinski criteria, from computed molecular properties of the geometry optimized structures. Computed descriptors often used to predict absorption, distribution, metabolism, elimination and toxicity (ADMET) were used to assess the pharmacokinetic profiles of the isolated compounds. Antiplasmodial activity was demonstrated for the first time in five major natural products previously identified in D. edulis, but not tested against malaria parasites. The most active compound identified was termed DES4. It had IC50 values of 0.37 and 0.55 µg/mL, against 3D7 and Dd2 respectively. In addition, this compound was shown to act in synergy with quinine, satisfied all criteria of "Drug-likeness" and showed considerable probability of providing an antimalarial lead. The remaining four compounds also showed antiplasmodial activity, but were less effective than DES4. None of the tested compounds was cytotoxicity against LLC-MK2 cells, suggesting their selective activities on malaria parasites. Based on the high in vitro activity, low toxicity and predicted "Drug-likeness" DES4 merits further investigation as a possible drug lead for the treatment of malaria.

Bioassay-guided discovery of antibacterial agents: in vitro screening of Peperomia vulcanica, Peperomia fernandopoioana and Scleria striatinux.

BACKGROUND: The global burden of bacterial infections is high and has been further aggravated by increasing resistance to antibiotics. In the search for novel antibacterials, three medicinal plants: Peperomia vulcanica, Peperomia fernandopoioana (Piperaceae) and Scleria striatinux (Cyperaceae), were investigated for antibacterial activity and toxicity. METHODS: Crude extracts of these plants were tested by the disc diffusion method against six bacterial test organisms followed by bio-assay guided fractionation, isolation and testing of pure compounds. The minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations were measured by the microdilution method. The acute toxicity of the active extracts and cytotoxicity of the active compound were performed in mice and mammalian cells, respectively. RESULTS: The diameter of the zones of inhibition (DZI) of the extracts ranged from 7-13 mm on Escherichia coli and Staphylococcus aureus of which the methylene chloride:methanol [1:1] extract of Scleria striatinux recorded the highest activity (DZI = 13 mm). Twenty-nine pure compounds were screened and one, Okundoperoxide, isolated from S. striatinux, recorded a DZI ranging from 10-19 mm on S. aureus. The MICs and MBCs indicated that the Peperomias had broad-spectrum bacteriostatic activity. Toxicity tests showed that Okundoperoxide may have a low risk of toxicity with an LC50 of 46.88 µg/mL. CONCLUSIONS: The antibacterial activity of these plants supports their use in traditional medicine. The pure compound, Okundoperoxide, may yield new antibacterial lead compounds following medicinal chemistry exploration.

Hypericum lanceolatum (Hypericaceae) as a potential source of new anti-malarial agents: a bioassay-guided fractionation of the stem bark.

BACKGROUND: Malaria is a major public health threat in Africa, and traditional medicine continues to play a key role in its control especially in rural areas. A bioassay-guided fractionation was carried out in order to evaluate the anti-malarial potential and the safety of the methanol extract of the Hypericum lanceolatum stem bark. METHODS: The anti-plasmodial activity was assayed by the lactate dehydrogenase method (pLDH) against the multidrug-resistant W2mef laboratory strain, and a field isolate (SHF4) of Plasmodium falciparum. Cytotoxicity tests were carried out using the LLC-MK2 monkey kidney epithelial cells. RESULTS: Five compounds were isolated from the most active and least cytotoxic ethylacetate sub-extract: betulinic acid (HLT1), 2,2',5,6'-tetrahydroxybenzophenone (HLT2), 5-hydroxy-3-methoxyxanthone (HLT3), 3-hydroxy-5-methoxyxanthone (HLT4) and HLT0 (yet to be identified). Three of the tested compounds presented significant anti-plasmodial activities (with 50% inhibitory concentration, IC50 < 5 µM), with 5-hydroxy-3-methoxyxanthone exerting the highest activity, followed by HLT0 and betulinic acid. All the compounds with significant anti-plasmodial activity were non-cytotoxic, except betulinic acid which showed a 50% cytotoxic concentration, CC50 of 25 µg/mL. CONCLUSIONS: These findings justify the use of H. lanceolatum stem bark as anti-malarial by traditional healers of Western Cameroon, and could constitute a good basis for further studies towards development of new drug candidates or phytomedicines for malaria.

In vitro antiplasmodial activity and cytotoxicity of crude extracts and compounds from the stem bark of Kigelia africana (Lam.) Benth (Bignoniaceae).

In order to assess the potential of the stem bark of Kigelia africana (Lam.) Benth as source of new anti-malarial leads, n-hexane and ethyl acetate (EtOAc) extracts and four compounds isolated from the stem bark were screened in vitro against the chloroquine-resistant W-2 and two field isolates of Plasmodium falciparum using lactate dehydrogenase assay. The products were also tested for their cytotoxicity on LLC/MK2 monkey kidney cells. The EtOAc extract exhibited a significant antiplasmodial activity (IC(50) = 11.15 µg/mL on W-2; 3.91 and 4.74 µg/mL on field CAM10 and SHF4 isolates, respectively), whereas the n-hexane fraction showed a weak activity (IC(50) = 73.78 µg/mL on W-2 and 21.85 µg/mL on SHF4). Three out of the four compounds showed good activity against all the three different parasite strains (IC(50) <5 µM). Specicoside exhibited the highest activity on W-2 (IC(50) = 1.54 µM) followed by 2ß, 3ß, 19α-trihydroxy-urs-12-en-28-oic acid (IC(50) = 1.60 µM) and atranorin (IC(50) = 4.41 µM), while p-hydroxycinnamic acid was the least active (IC(50) =53.84 µM). The EtOAc extract and its isolated compounds (specicoside and p-hydroxycinnamic acid) were non-cytotoxic (CC(50) > 30 µg/mL), whereas the n-hexane extract and two of its products, atranorin and 2ß, 3ß, 19α-trihydroxy-urs-12-en-28-oic acid showed cytotoxicity at high concentrations, with the last one being the most toxic (CC(50) = 9.37 µg/mL). These findings justify the use of K. africana stem bark as antimalaria by traditional healers of Western Cameroon, and could constitute a good basis for further studies towards development of new leads or natural drugs for malaria.

In vitro antiplasmodial activity and cytotoxicity of extracts of selected medicinal plants used by traditional healers of Western cameroon.

Medicinal plants play a key role in malaria control in Africa, especially in remote areas where health facilities are limited. In order to assess their acclaimed potentials, eleven extracts were prepared from seven selected plants commonly used in Western Cameroon, and tested both for their antiplasmodial activity and cytotoxicity. The antiplasmodial activity was assessed using Lactate Dehydrogenase Assay (pLDH) and the cytotoxicity estimated on LLC-MK2 monkey kidney epithelial cells. Seven extracts from five different plants were significantly active, with very weak or no cytotoxicity. The Dacryodes edulis leaves showed the highest activity (IC(50) of 6.45 µg/mL on 3D7 and 8.2 µg/mL on DD2) followed by the leaves of Vernonia amygdalina (IC(50) of 8.72 and 11.27 µg/mL on 3D7 and DD2 resp.) and roots of V. amygdalina (IC(50) of 8.72 µg/mL on 3D7), Coula edulis leaves (IC(50) of 13.80 µg/mL and 5.79 µg/mL on 3D7 and DD2 resp.), Eucalyptus globulus leaves (IC(50) of 16.80 µg/mL and 26.45 µg/mL on 3D7 and DD2) and Cuviera longiflora stem bark (IC(50) of 20.24 µg/mL and 13.91 µg/mL on 3D7 and DD2). These findings justify the use of five of the seven plants in malaria treatment by traditional healers of Western Cameroon.

Selective activity of extracts of Margaritaria discoidea and Homalium africanum on Onchocerca ochengi.

BACKGROUND: The current treatment of onchocerciasis relies on the use of ivermectin which is only microfilaricidal and for which resistant parasite strains of veterinary importance are increasingly being detected. In the search for novel filaricides and alternative medicines, we investigated the selective activity of crude extracts of Margaritaria discoidea and Homalium africanum on Onchocerca ochengi, a model parasite for O. volvulus. These plants are used to treat the disease in North West Cameroon. METHODS: Sixteen crude extracts were prepared from various parts of M. discoidea and H. africanum using different organic solvents. The filaricidal activities were determined in vitro. Cytotoxicity of the active extracts was assessed on monkey kidney epithelial cells in vitro and the selectivity indices (SI) of the extracts determined. Acute toxicity of the promising extracts was investigated in mice. RESULTS: Four out of the 16 extracts showed microfilaricidal activity based on motility reduction, whereas, none showed macrofilaricidal activity based on the MTT/formazan assay. The methylene chloride extract of H. africanum leaves (HLC) recorded the lowest IC50 of 31.25 µg/mL and an IC100 of 62.5 µg/mL. The SI for the active extracts ranged from 0.5 - 2.63. No form of acute toxicity was observed in mice. Phytochemical analysis revealed the presence of anthraquinones, sterols and terpenoids in the promising extracts. CONCLUSIONS: The non-polar extracts of M. discoidea and H. africanum are potential sources of new microfilaricidal lead compounds, and the results support their use in traditional medicine.

The antimalarial potential of medicinal plants used for the treatment of malaria in Cameroonian folk medicine.

Malaria remains one of the leading public health problems in Cameroon as in other parts of Sub-Saharan Africa. In the past decades, this situation has been aggravated by the increasing spread of drug-resistant Plasmodium falciparum strains. New antimalarial drug leads are therefore urgently needed. Traditional healers have long used plants to prevent or cure infections. This article reviews the current status of botanical screening efforts in Cameroon as well as experimental studies done on antimalarial plants. Data collected from 54 references from various research groups in the literature up to June 2007 shows that 217 different species have been cited for their use as antimalarials in folk medicine in Cameroon. About a hundred phytochemicals have been isolated from 26 species some among which are potential leads for development of new antiamalarials. Crude extracts and or essential oils prepared from 54 other species showed a wide range of activity on Plasmodium spp. Moreover, some 137 plants from 48 families that are employed by traditional healers remain uninvestigated for their presumed antimalarial properties. The present study shows that Cameroonian flora represents a high potential for new antimalarial compounds. Further ethnobotanical surveys and laboratory investigations are needed to fully exploit the potential of the identified species in the control of malaria.

Antiplasmodial activities of some products from Turreanthus africanus (Meliaceae).

We investigated the antiplasmodial activity of some pure compounds of Turreanthus africanus (Meliaceae), a plant that is used in traditional medicine to treat malaria in Southwest Cameroon. A phytochemical analysis of the methylene chloride: methanol (1:1) extract of the seeds of the plant yielded seven compounds. Four of them, which were oils, were subjected to in vitro bioassays on Plasmodium falciparum F 32, chloroquine sensitive strain. Compound 1 (16-oxolabda-8 (17), 12(E)-dien-15-oic acid), showed the highest antiplasmodial activity, two others (methyl-14,15-epoxylabda-8 (17), 12(E)-diene-16-oate, and turreanin A), had moderate activity and one was inactive. These findings are consistent with the use of T. africanus in the traditional treatment of P. falciparum malaria.