Antimicrob Agents Chemother 48(8): 3010-3015

Possible Mechanism of Miltefosine-Mediated Death of <em>Leishmania donovani</em><sup><a href="#fn1" rid="fn1" class=" fn">†</a></sup>

Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Punjab, India¹

^{Corresponding author. Mailing address: Department of Biotechnology, NIPER, Sector 67, S.A.S. Nagar, Punjab 160 062, India. Phone: 91(0)172-214682. Fax: 91(0)172-214682. E-mail:}ni.ca.repin@yedsc.

Received 2004 Feb 10; Revised 2004 Mar 29; Accepted 2004 Apr 5.

Abstract

Miltefosine causes leishmanial death, but the possible mechanism(s) of action is not known. The mode of action of miltefosine was investigated in vitro in Leishmania donovani promastigotes as well as in extra- and intracellular amastigotes. Here, we demonstrate that miltefosine induces apoptosis-like death in L. donovani based on observed phenomena such as nuclear DNA condensation, DNA fragmentation with accompanying ladder formation, and in situ labeling of DNA fragments by the terminal deoxyribonucleotidyltransferase-mediated dUTP-biotin nick end labeling method. Understanding of miltefosine-mediated death will facilitate the design of new therapeutic strategies against Leishmania parasites.

Abstract

Miltefosine (1-O-hexadecylphosphocholine), an alkylphosphocholine and a membrane-active synthetic ether-lipid analogue originally developed for the treatment of cutaneous metastasis from mammary carcinomas (21, 32), has proved to be an effective treatment for human visceral leishmaniasis (25, 38, 52-55). It has been hailed as potentially the first oral treatment of human leishmaniasis (8, 15, 16, 20, 25, 31, 47, 50). The leishmaniacidal activities of miltefosine have been associated with perturbation of the alkyl-phospholipid metabolism and the biosynthesis of alkyl-anchored glycolipids and glycoproteins (33, 34). Although potential antitumor cell mechanisms of action of miltefosine have been elaborated in mammalian cells (13, 44), its exact mode(s) of cytotoxicity has not been determined in Leishmania spp. (9, 25, 39, 46, 48, 55). It has been known to induce apoptotic death in various cancer cell lines (13, 28, 32, 43, 60). However, it is not yet established whether miltefosine can bring about apoptosis-like death in all the forms of Leishmania parasite.

There are now increasing numbers of reports regarding unicellular organisms undergoing apoptosis-like death, whose induction is not obligatory but activated under threatening circumstances (1, 36, 49). Cell death resembling metazoan apoptosis has been reported in several parasitic protozoans (4, 10, 30, 36, 49, 59, 61). Apoptosis greatly affects the host-parasite relationship, since the survival of the parasite inside the vector as well as in the macrophage requires strict control of the population of the parasite (12, 58). Apoptosis could be a useful mechanism to avoid killing of the entire population (36) and thus influence the chemotherapeutic strategies to limit the parasite (11).

In the present study we sought to determine the mode of action of miltefosine in Leishmania donovani promastigotes as well as extra- and intracellular amastigotes. We have demonstrated that miltefosine causes apoptosis-like death in L. donovani. Our data set the stage for future development of this class of drug for better treatment of leishmaniasis.

Acknowledgments

We thank C. L. Kaul, NIPER, for his keen interest in this study. We thank R. Mahajan for providing us with the Leishmania donovani strain MHOM/80/IN/Dd8 and S. S. Sharma for providing the peritoneal macrophages and amastigotes used in this study. We acknowledge K. G. Jayanarayan and A. Khurana for their support in executing some experiments and helpful discussions. R. Singh is acknowledged for his assistance in the laboratory.

N.K.V. is recipient the of a Research Associateship from NIPER.

Acknowledgments

Footnotes

^{NIPER communication no. 288.}

Footnotes

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