Miltefosine-Induced Apoptotic Cell Death onLeishmania majorandL. tropicaStrains

Abstract

The aim of this study was to assess the cytotoxic effects of various concentrations of miltefosine on Leishmania major (MRHO/IR/75/ER) and L. tropica (MHOM/IR/02/Mash10) promastigotes and to observe the programmed cell death features. The colorimetric MTT assay was used to find L. major and L. tropica viability and the obtained results were expressed as 50% inhibitory concentration (IC50). Also, 50% effective doses (ED50) for L. major and L. tropica amastigotes were also determined. Annexin-V FLUOS staining was performed to study the cell death properties of miltefosine using FACS analysis. Qualitative analysis of the total genomic DNA fragmentation was performed by agarose gel electrophoresis. Furthermore, to observe changes in cell morphology, promastigotes were examined using light microscopy. In both strains of L. major and L. tropica, miltefosine induced dose-dependent death with features of apoptosis, including cell shrinkage, DNA laddering, and externalization of phosphatidylserine. The IC50 was achieved at 22 µM and 11 µM for L. major and L. tropica after 48 hr of incubation, respectively. ED50 of L. major and L. tropica amastigotes were 5.7 µM and 4.2 µM, respectively. Our results indicate that miltefosine induces apoptosis of the causative agent of cutaneous leishmaniasis in a dose-dependent manner. Interestingly, L. major did not display any apoptotic changes when it was exposed to miltefosine in concentrations sufficient to kill L. tropica.

Leishmania Promastigotes Lack Phosphatidylserine but Bind Annexin V upon Permeabilization or Miltefosine Treatment

“…Nevertheless, several of these parasite species have been reported to bind annexin V under certain conditions [16], [30], [31], [32]. In the present report, we observed strong staining of L. donovani promastigotes by annexin V upon electroporation or miltefosine treatment despite the absence of detectable levels of PS.…”

Utility of amino acid coupled 1,2,4-triazoles in organic synthesis: synthesis of some new antileishmainal agents

“…The common form of leishmaniasis is Cutaneous leishmaniasis (CL) which widely distributed all around the world [3]. Within the last decade, the treatment is limited to a few drugs, such as amphotericin B, miltefosine and paromomycin and far from satisfactory [4], although a broad array of species can be responsible to cause leishmaniasis, affecting humans and animals [6][7][8][9][10][11]. Miltefosine-resistant Leishmania donovani promastigotes also demonstrated modification in sterol biosynthesis and lipid compositions which influences the membrane fluidity and permeability and ultimately may affect drug-membrane interactions [5].…”

Molecular docking and molecular dynamics simulation study of inositol phosphorylceramide synthase – inhibitor complex in leishmaniasis: Insight into the structure based drug design

“…Newer formulations for the treatment of this disease include the administration of miltefosine. Miltefosine (hexadecylphosphocholine), originally an anticancer drug has been reported to induce apoptosis of L. major amastigotes in the infected macrophages 3 . However the growing problem of drug resistance to the existing chemotherapeutics as well as the quick adaptability of the parasite to the host immune responses has necessitated the development of newer treatment strategies for leishmaniasis 4– 6 .…”