Pharmaceutics Division, CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, UP, India.
Parasitology Division, CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, UP, India.
Int J Biol Macromol. 2017 Dec;105(Pt 1):1220-1231. doi: 10.1016/j.ijbiomac.2017.07.161. Epub 2017 Aug 3.
The goal of study was to develop micellar formulation of Amphotericin B (AmB) to improve its antileishmanial efficacy. AmB loaded pluronic F127 (PF 127) micelles were developed and coated with chitosan (Cs-PF-AmB-M) to accord immunoadjuvant and macrophage targeting properties. Hemolysis and cytotoxicity studies demonstrated that Cs-PF-AmB-M was 7.93 fold (at 20μg/ml AmB concentration) and 9.35 fold less hemolytic and cytotoxic, respectively in comparison to AmB suspension. Flow cytometry studies indicated that Cs-PF-FITC-M was 21.97 fold higher internalized byJ774A.1 macrophage in comparison to PF-FITC-M.Cs-PF-AmB-M showed excellent in-vitro (1.82 fold in compared to AmB suspension) and in-vivo (75.84±7.91% parasitic inhibition) antileishmanial activity against macrophage resident intracellular promastigotes and Leishmania donovani infected Syrian hamsters, respectively. Chitosan coating stimulated a Th1 immune response mediating auxiliary immunotherapeutic action as judged by in-vitro and in-vivo cytokine and mRNA expression. Toxicity studies demonstrated normal blood urea nitrogen (BUN) and plasma creatinine (PC) level and no sign of abnormal histopathology upon intravenous administration of micellar formulations. Pharmacokinetic profiling and tissue distribution studies indicated that AmB was preferentially localized in macrophage harboring tissue instead of kidney, thereby circumventing the characteristic nephrotoxicity. Conclusively, Cs-PF-AmB-M could be a viable alternative for the current immuno and chemotherapy of visceral leishmaniasis (VL).
本研究旨在开发两性霉素 B(AmB)胶束制剂以提高其抗利什曼原虫的疗效。研制了两性霉素 B 载 Pluronic F127(PF 127)胶束(Cs-PF-AmB-M),并对其进行壳聚糖包被,赋予免疫佐剂和巨噬细胞靶向特性。溶血和细胞毒性研究表明,与 AmB 混悬剂相比,Cs-PF-AmB-M 的溶血率和细胞毒性分别降低了 7.93 倍(在 20μg/ml AmB 浓度下)和 9.35 倍。流式细胞术研究表明,与 PF-FITC-M 相比,Cs-PF-FITC-M 被 J774A.1 巨噬细胞内化的效率高 21.97 倍。Cs-PF-AmB-M 表现出优异的体外(与 AmB 混悬剂相比提高了 1.82 倍)和体内(对巨噬细胞内驻留前鞭毛体和感染的叙利亚仓鼠利什曼原虫的寄生虫抑制率分别为 75.84±7.91%)抗利什曼原虫活性。壳聚糖包被刺激 Th1 免疫反应,通过体外和体内细胞因子和 mRNA 表达判断具有辅助免疫治疗作用。毒性研究表明,胶束制剂静脉给药后,血尿素氮(BUN)和血浆肌酐(PC)水平正常,无组织病理学异常迹象。药代动力学特征分析和组织分布研究表明,AmB 优先定位于含有巨噬细胞的组织中,而不是肾脏,从而避免了特征性的肾毒性。总之,Cs-PF-AmB-M 可能是治疗内脏利什曼病(VL)的现有免疫和化疗的可行替代方案。