IN-VITRO ANTILEISHMANIAL EVALUATION OF AMPHOTERICIN B-MILTEFOSINE NANOVESICLES

Leishmaniasis is a neglected tropical infectious disease affecting poor people, especially in the developing countries of the world. It is a systemic disease caused by a protozoan parasite leishmania donovani and is fatal if left untreated. The aim of the present study is to develop and evaluate the in-vitro antileishmanial activity of the combination of amphotericin B and mitefosinenano vesicles for targeted delivery to macrophages in the treatment of visceral leishmaniasis to increase the efficacy, decrease emergency of resistance and increase overall patient compliance. Drug-loaded nanovesicles were prepared by ethanol injection method. Physicochemical characterization and cytotoxicity, and antileishmanial activity evaluations were assessed. Nanovesicles of optimal AmB-MTF formulation was prepared from phosphatidylcholine-cholesterol-stearic acid (20:4:1 w/w) lipids, ethanol-water (1:4 v/v) dispersion medium, and AmB-MTF (1:1 w/w) drugs at drug loading of 1:8 and stirring rate of 1000 rpm. AmB-MTF 1:1 nanovesicles exhibited a mean particle size of 145.6 nm, polydispersity index (PDI) 0.19, zeta potential -27.3 mV and drug entrapment efficiency 87%. The AmB-MTF combination nanovesicles demonstrated low CC50 against J774 A.1 murine macrophage cells (18.12 μM) compared to the plain AmB (3.71μM), AmB nanovesicles (13.61μM) or AmB-MTF physical mixture (13.64  μM). In-vitro antileishmanial activity against promastigotes study indicated that IC⁵⁰ of AmB-MTF1:1 nanovesicles (0.046) was significantly decreased (p<0.05) compared to plain AmB-MTF mixture (0.063) or AmB nanovesicles (0.055). To conclude, AmB-MTF nanovesicles could be a safe and reliable therapeutic option over conventional AmB-MTF combination therapy with due consideration in-vivo evaluations to be investigated.