Jain Keerti, Verma Ashwni Kumar, Mishra Prabhat Ranjan, Jain Narendra Kumar
Pharmaceutics Research Laboratory, Department of Pharmaceutical Sciences, Dr. H. S. Gour Central University, Sagar, India Pharmaceutical Nanotechnology Laboratory, ISF College of Pharmacy, Moga, India
Pharmaceutics Division, CSIR-Central Drug Research Institute, Lucknow, India.
Antimicrob Agents Chemother. 2015 May;59(5):2479-87. doi: 10.1128/AAC.04213-14. Epub 2015 Feb 2.
The present study aimed to develop an optimized dendrimeric delivery system for amphotericin B (AmB). Fifth-generation (5.0 G) poly(propylene imine) (PPI) dendrimers were synthesized, conjugated with mannose, and characterized by use of various analytical techniques, including Fourier transform infrared spectroscopy (FTIR), (1)H nuclear magnetic resonance ((1)H-NMR) spectroscopic analysis, and atomic force microscopy (AFM). Mannose-conjugated 5.0 G PPI (MPPI) dendrimers were loaded with AmB and evaluated for drug loading efficiency, in vitro drug release profile, stability, hemolytic toxicity to human erythrocytes, cytotoxicity to and cell uptake by J774A.1 macrophage cells, antiparasitic activity against intracellular Leishmania donovani amastigotes, in vivo pharmacokinetic and biodistribution profiles, drug localization index, toxicity, and antileishmanial activity. AFM showed the nanometric size of the MPPI dendrimers, with a nearly globular architecture. The conjugate showed a good entrapment efficiency for AmB, along with pH-sensitive drug release. Highly significant reductions in toxicity toward human erythrocytes and macrophage cells, without compromising the antiparasitic activity of AmB, were observed. The dendrimeric formulation of AmB showed a significant enhancement of the parasiticidal activity of AmB toward intramacrophagic L. donovani amastigotes. In the in vitro cell uptake studies, the formulation showed selectivity toward macrophages, with significant intracellular uptake. Further pharmacokinetic and organ distribution studies elucidated the controlled delivery behavior of the formulation. The drug localization index was found to increase significantly in macrophage-rich organs. In vivo studies showed a biocompatible behavior of MPPIA, with negligible toxicity even at higher doses, and promising antileishmanial activity. From the results, we concluded that surface-engineered dendrimers may serve as optimized delivery vehicles for AmB with enhanced activity and low or negligible toxicity.
本研究旨在开发一种用于两性霉素B（AmB）的优化树枝状递送系统。合成了第五代（5.0 G）聚（丙烯亚胺）（PPI）树枝状大分子，将其与甘露糖共轭，并使用各种分析技术进行表征，包括傅里叶变换红外光谱（FTIR）、氢核磁共振（¹H-NMR）光谱分析和原子力显微镜（AFM）。用AmB负载甘露糖共轭的5.0 G PPI（MPPI）树枝状大分子，并评估其载药效率、体外药物释放曲线、稳定性、对人红细胞的溶血毒性、对J774A.1巨噬细胞的细胞毒性和细胞摄取、对细胞内杜氏利什曼原虫无鞭毛体的抗寄生虫活性、体内药代动力学和生物分布曲线、药物定位指数、毒性和抗利什曼活性。AFM显示了MPPI树枝状大分子的纳米尺寸，具有近乎球状的结构。该共轭物对AmB显示出良好的包封效率，以及pH敏感的药物释放。观察到对人红细胞和巨噬细胞的毒性显著降低，同时不影响AmB的抗寄生虫活性。AmB的树枝状制剂显示出AmB对巨噬细胞内杜氏利什曼原虫无鞭毛体的杀寄生虫活性显著增强。在体外细胞摄取研究中，该制剂对巨噬细胞具有选择性，细胞内摄取显著。进一步的药代动力学和器官分布研究阐明了该制剂的控释行为。发现药物定位指数在富含巨噬细胞的器官中显著增加。体内研究显示MPPIA具有生物相容性，即使在高剂量下毒性也可忽略不计，并且具有良好的抗利什曼活性。从结果中，我们得出结论，表面工程化的树枝状大分子可作为AmB的优化递送载体，具有增强的活性和低毒性或可忽略不计的毒性。
