title: Nanonization increases the antileishmanial efficacy of amphotericin B: an ex vivo approach.
abstract: With widespread resistance to pentavalent antimonial in the endemic eastern terai belt of Nepal and Bihar, India, Amphotericin B deoxycholate is now the first-line antileishmanial drug for the treatment of visceral leishmaniasis (VL). However, universal occurrence of infusion-related fever and rigors with amphotericin B (AmB), occasional serious life-threatening toxicities like cardiotoxicity, anaphylaxis, hypokalemia, and nephrotoxicity are major barriers to its use in areas with limited medical facilities. Liposomal amphotericins, however, are devoid of adverse effects, high cost makes it unaffordable. We had formulated nanoparticles (10-20 nm) from amphotericin B deoxycholate (1-2 μm) applying high pressure (150 atm) milling homogenization in argon atmosphere and tested its ex vivo efficacy in Leishmania infected J774A cell line and peritoneal macrophage. The ex vivo ED50 for intracellular amastigotes in peritoneal macrophage by nano-amphotericin was 0.0027 ± 0.001 μg/mL which was significantly less (p = 0.0029) than the required dose of amphotericin B (0.0426 ± 0.003 μg/mL). Similarly, in J774A cell line, 50 % of intracellular amastigotes were cleared by 0.0038 ± 0.001 μg/mL of nano-amphotericin while the dose was a bit more for AmB (0.0196 ± 0.001 μg/mL) illustrating the significant difference (p value, 0.0122). The nanoformulation has also shown high efficacy (ED50, 0.0028-0.0035 μg/mL) in inhibition of infected macrophage count. The new formulation accumulated to spleen, the targeted organ, 7 days after inoculation of drug to the infected hamster as traced in vivo by TEM convincing it as potential drug. Given a favorable safety profile and very low cost of production contemplated, it may prove to be a feasible alternative for conventional amphotericin B.
authors.affiliation: Central Department of Biotechnology, Institute of Science and Technology, Tribhuvan University, Kathmandu, Nepal, krishna.manandhar@gmail.com.
authors.forename: Krishna Das
authors.initials: KD
authors.lastname: Manandhar
authors.forename: Thakur Prasad
authors.initials: TP
authors.lastname: Yadav
authors.forename: Vijay Kumar
authors.initials: VK
authors.lastname: Prajapati
authors.forename: Om
authors.initials: O
authors.lastname: Basukala
authors.forename: Ram Prasad
authors.initials: RP
authors.lastname: Aganja
authors.forename: Anuradha
authors.initials: A
authors.lastname: Dude
authors.forename: Onkar Nath
authors.initials: ON
authors.lastname: Shrivastav
authors.forename: Shyam
authors.initials: S
authors.lastname: Sundar
mesh_terms: D000666:Amphotericin B / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D000981:Antiprotozoal Agents / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D006224:Cricetinae; D003840:Deoxycholic Acid / Q000737:chemistry / Q000494:pharmacology*; D004338:Drug Combinations; D054796:Drug Dosage Calculations; D006801:Humans; D020128:Inhibitory Concentration 50; D007893:Leishmania donovani / Q000187:drug effects* / Q000254:growth & development / Q000472:pathogenicity; D007898:Leishmaniasis, Visceral / Q000188:drug therapy* / Q000469:parasitology / Q000662:veterinary*; D008018:Life Cycle Stages / Q000187:drug effects / Q000502:physiology; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000469:parasitology; D051379:Mice; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry* / Q000648:ultrastructure; D013154:Spleen / Q000187:drug effects / Q000469:parasitology
chemical_list: D000981:Antiprotozoal Agents; D004338:Drug Combinations; D003840:Deoxycholic Acid; D000666:Amphotericin B; C059765:amphotericin B, deoxycholate drug combination