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Nanonization Increases the Antileishmanial Efficacy of Amphotericin B: An Ex Vivo Approach

  • Krishna Das ManandharEmail author
  • Thakur Prasad Yadav
  • Vijay Kumar Prajapati
  • Om Basukala
  • Ram Prasad Aganja
  • Anuradha Dude
  • Onkar Nath Shrivastav
  • Shyam Sundar
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 808)

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.

Keywords

Amphotericin B Visceral leishmaniasis Leishmania donovani Nanoparticles Hamster Ex vivo 

Notes

Acknowledgments

The authors are thankful to the Nano Science and Technology Initiative, Department of Science and Technology (DST), India for financial support in the nanonization part. We gratefully acknowledge Kala-azar Medical Research Centre, Muzaffarpur, India for providing infected aspirates to culture the parasites. Prof. Manandhar is thankful to UGC, Nepal for providing fellowship to him during the study.

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Copyright information

© Springer India 2014

Authors and Affiliations

  • Krishna Das Manandhar
    • 1
    • 2
    Email author
  • Thakur Prasad Yadav
    • 3
  • Vijay Kumar Prajapati
    • 2
    • 4
  • Om Basukala
    • 1
    • 5
  • Ram Prasad Aganja
    • 1
  • Anuradha Dude
    • 6
  • Onkar Nath Shrivastav
    • 3
  • Shyam Sundar
    • 2
  1. 1.Central Department of Biotechnology, Institute of Science and TechnologyTribhuvan UniversityKathmanduNepal
  2. 2.Infectious Disease Research Laboratory, Department of Medicine, Institute of Medical ScienceBanaras Hindu UniversityVaranasiIndia
  3. 3.Department of PhysicsBanaras Hindu UniversityVaranasiIndia
  4. 4.Department of BiochemistryThe Central University of RajasthanAjmerIndia
  5. 5.Nepal Academy of Science and Technology (NAST)LalitpurNepal
  6. 6.Central Drug Research InstituteLucknowIndia

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