Antitrypanosomal properties of extracts and their inhibitory effect on trypanosome alternative oxidase.

BACKGROUND

African trypanosomiasis is a protozoan disease with huge socio-economic burden to sub-Saharan African exceeding US$4.6 annual loss. To mitigate the incidence of trypanosomal drug resistance, efforts are geared towards discovery of molecules, especially from natural products, with potential to inhibit important molecular target (trypanosome alternative oxidase, TAO) in trypanosomes that are critical to their survival.

METHOD

Crude methanol extract of was subjected to bioassay-guided antitrypanosomal assay to identify the most active extract with trypanocidal activity. The most active extract was run on a column chromatography yielding five fractions, F1-F5. The fractions were assayed for inhibitory effect on TAO. The most promising TAO inhibitor was subjected to antitrypanosomal evaluation by trypanosome count, drug incubation infectivity test (DIIT) and in vivo studies. Gas chromatography-mass spectrometry (GC-MS) was used to identify and quantify phytochemical constituents of the potential TAO-inhibiting fraction.

RESULTS

Ethyl acetate extract (EtOAc) significantly (<0.05) produced trypanocidal effect and was the most active extract. Of the five fractions, only F4 significantly (<0.05) inhibited TAO compared to the control. F4 completely immobilised the trypanosomes up to 0.5 μg/μl, yielding an EC of 0.024 μg/μl compared to the 0.502 μg/μl of diminazene aceturate positive control group. The DIIT showed that F4 was significantly (<0.05) potent up to 0.1 μg/μl. F4 significantly (<0.05) suppressed parasite multiplication in systemic circulation of the treated rats and significantly (<0.05) maintained high PCV when compared to the 5% DMSO group. Furthermore, F4 significantly (<0.05) lowered serum concentrations of malondialdehyde. Phytoconstituents identified by the GC-MS include tetradecene; cetene; 3-(benzylthio) acrylic acid, methyl ester; 1-octadecene; 9-heptadecanone; hexadecanoic acid, methyl ester; dibutyl phthalate; eicosene; octadecenoic acid, methyl ester; oleic acid; 2-methyl-Z,Z-3,13-octadecadienol; 1-docosene; 3-phenylthiane, s-oxide; phenol, 3-methyl; phthalic acid, di(2-propylpentyl) ester and 1,4-benzenedicarboxylic acid, bis (2-ethylhexyl) ester.

CONCLUSION

F4 from EtOAc contains six carbohydrates (9.58%), two free fatty acids (6.48%), five fatty acid esters (27.73%), two aromatic compounds (50.63%) and one organosulphide (5.61%). It inhibited TAO and demonstrated antitrypanosomal effects.