Amphotericin B-cholesterol Co-encapsulated Polymeric Nanoparticles in Protein Hydrogel-based Injectable Depot System for Long-acting Drug Delivery.

Amphotericin B exhibits dose-dependent nephrotoxicity and hepatotoxicity due to its non-selective interaction with mammalian cholesterol. The aim of the present study was to develop an Amphotericin B-cholesterol co-encapsulated albumin hydrogel-based injectable depot formulation to achieve prolonged plasma drug concentrations with reduced toxicity. Amphotericin B-loaded polymeric nanoparticles (Amphotericin B-PNs) were prepared by the nanoprecipitation method. Cholesterol co-encapsulated nanoformulation has been able to keep Amphotericin B in a less aggregated state (aggregation ratio -1.40), exhibited less hemolysis (< 10%) in rat erythrocytes, higher cytocompatibility in HEK 293 T and RAW 264.7 cell lines, as well as time-dependent increased intracellular accumulation of C6 loaded nanoparticles. Amphotericin B-PN-loaded albumin hydrogel was prepared by an ethanol-induced gelation process. Nanoparticle-loaded hydrogel exhibited sustained drug release, and at the end of one week ~ 60% of Amphotericin B was released. Hydrogel formulations have shown improved pharmacokinetic parameters with prolonged drug release till the end of the 14th day. The cholesterol co-encapsulated hydrogel has also shown reduced organ toxicity and improved safety against oxidative stress, as revealed by biochemical (creatinine, BUN, ALT, and AST) and ROS (GSH, catalase, NO, and MDA) scavenging assays. Overall, it can be concluded that the Amphotericin B-PN-loaded albumin hydrogel could be a potential delivery system for the long-acting release of Amphotericin B with reduced toxicity.