Long-term glycemic control and prevention of diabetes complications in vivo using oleic acid-grafted-chitosan‑zinc-insulin complexes incorporated in thermosensitive copolymer

Abstract

Daily injections for basal insulin therapy are far from ideal resulting in hypo/hyperglycemic episodes associated with fatal complications in type-1 diabetes patients. Here we report a delivery system that provides controlled release of insulin closely mimicking physiological basal insulin requirement for an extended period following a single subcutaneous injection. Stability of insulin was significantly improved by formation of zinc-insulin hexamers, further stabilized by electrostatic complex formation with chitosan polymer. Insulin complexes were homogenously incorporated into PLA-PEG-PLA, a biodegradable thermogel copolymer, that instantaneously forms a subcutaneous gel-depot following injection. Chitosan polymer was hydrophobically modified using oleic acid prior to complex formation with insulin to enable distribution of oleic acid-grafted-chitosan‑zinc-insulin complexes into the hydrophobic core of PLA-PEG-PLA thermogel-copolymer micelles. In vivo, daily administration of marketed long-acting insulin, glargine, resulted in fluctuating blood glucose levels between 91 and 443 mg/dL in type 1 diabetic rats. However, single administration of thermogel copolymeric formulation successfully demonstrated slow diffusion of insulin complexes maintaining peak-free basal insulin level of 21 mU/L for 91 days. Sustained release of basal insulin also correlated with efficient glycemic control (blood glucose <120 mg/dL), prevention of diabetic ketoacidosis and absence of cataract development, unlike other treatment groups. Moreover, there was no sign of inflammation, tissue damage, or collagen deposition around depot site, suggesting exceptional biocompatibility of the formulation for long-term use.