Leishmanicidal activity of edelfosine, miltefosine and ilmofosine

Abstract

The anti-proliferative action of three alkyl-lysophospholipid derivatives, edelfosine (ET-OCH), miltefosine (Hexadecylphosphocholine), and ilmofosine (BM 14.440) has been studied on the promastigotes and amastigotes of Leishmania donovani. The effect of the three drugs has previously been studied, but the action mode was not clearly elucidated. In this study the effect on the intracellular amastigote forms was evaluated by two different methods: the traditional method, counting the amastigotes within the macrophages stained with Giemsa; and by a new method, staining the nuclear macrophages and amastigotes with ethidium bromide and counting the different population by flow cytometry. This new method, based on the flow cytometry, shows an advantage for evaluating the anti-proliferative effects in intracellular parasites. The ED50 were calculated for the drug activity after 72 hr, and for the three alkyl-lysophospholipid derivatives it were in the range of 26.73-33.31 microM against promastigotes and in the range of 16.46-23.16 against amastigotes. Also, studying the effect against macrophages J774A1, the ED50 were in the range of 24.28-26.38 microM. The effect of the alkyl-lysophospholipids in the macromolecular biosynthesis of the Leishmania donovani, was studied comparing the incorporation of labelled analogues ([3H] thymidine, [3H] uridine and [3H] leucine), respectively, in the DNA, RNA, and proteins of the flagellates treated. Miltefosine was the most active of the alkyl-lysophospholipids, especially in the inhibition of the RNA synthesis. The three compounds studied show high in vitro activity against L. donovani promastigotes and amastigotes.