Enhancing Safety and Efficacy by Altering the Toxic Aggregated State of Amphotericin B in Lipidic Nanoformulations.

Abstract

The toxicity of Amphotericin B (AmB) is contributed by the small, water-soluble aggregates of the drug. Hence, AmB lipid polymer hybrid nanoparticles (LIPOMER), comprising stearate lipids with a hydrophilic polymer Gantrez (GZ), and solid lipid nanoparticles (SLN), comprising only stearates, were prepared with the objective of monomerizing AmB. While intercalation of stearates with the hydrophobic polyene chain could hinder AmB-AmB interactions, enabling monomerization, it was hypothesized that GZ could aid in the stabilization of the monomers through hydrophilic interactions. AmB LIPOMERs and SLNs, prepared by nanoprecipitation, exhibited an average size of 350-500 nm with negative ζ potential. Polyglyceryl-6-distearate (PGDS) SLN exhibited maximum monomerization, with the highest peak IV (410 nm) to peak I (350 nm) ratio in the UV-visible spectrum. In total contrast, LIPOMERs and GZ nanoparticles revealed a hypsochromic shifted peak I between 321 and 324 nm, indicative of AmB super-aggregateformation. Super-aggregates, which result due to condensation of multiple aggregates with monomers, were attributed to extensive GZ-AmB and GZ-GZ interactions and could provide advantages of enhanced thermodynamic stability, with safety and efficacy similar to the monomeric form. Safety was confirmed by low and comparable erythrocyte toxicity exhibited by the LIPOMERs and SLNs. Anin vitro efficacy study of PGDS LIPOMER and SLN against intracellular amastigotes revealed significantly lower IC50 values, which translated to a 7.1- and 6.1-fold enhancement in efficacy compared to commercial nanoformulations Amfocare (micellar AmB) and 1.79- and 1.54-fold enhancement in efficacy compared to Fungisome (liposomal AmB). High efficacy coupled with a higher selectivity index indicated the superiority of the developed AmB nanoformulations and substantiated that altering the toxic aggregated state of AmB can offer a promising approach for the design of safe and efficacious AmB lipidic nanoformulations.