Glycosaminoglycan-Conjugated Insulin Derivatives Suitable for Once-Daily Formulations

Abstract

A novel, long-acting insulin conjugate was developed by modification with a glycosaminoglycan (GAG; i.e., chondroitin [CH] or heparosan [HPN]) at different positions (GlyA1, LysB29, or both) via different linkers with varying arm lengths. The in vitro glucose uptake-enhancing activity of conjugates inversely correlated with the circulatory in vivo half-life in mice and was affected primarily by the conjugation position rather than linker arm length. Conjugation at GlyA1, which provided the best balanced profile of in vitro activity and circulation period in mice, also provided the strongest glucose-lowering efficacy, exhibiting 12 h durability, which was superior to that of insulin glargine and comparable to that of insulin degludec in streptozotocin (STZ)-treated mice. The use of different GAGs did not significantly affect blood circulation or efficacy in mice; however, blood levels of CH-conjugated insulin and efficacy were lower in rats, indicating species differences in the performance of CH. Finally, optimal activity was obtained by conjugation with HPN at GlyA1 using a C3 linker. These results demonstrate the applicability of GAG modification for prolonging the efficacy of peptide drugs, similar to our previous application to glucagon-like peptide 1.

A glycosaminoglycan microarray identifies the binding of SARS‐CoV‐2 spike protein to chondroitin sulfate E

“…Biotinylated GAG was conjugated with BSA as previously described, but with some modifications [ 14 ]. Aminoalkanethiol (40 mg/mL) dissolved in deionized water was mixed with 50 mg/mL biotinylated GAG and sodium cyanoborohydride solution (Tokyo Chemical Industry Co., Ltd., Tokyo, Japan) and reacted under dark conditions at 43 °C for 16 h. The SH‐modified GAG that was generated at the reducing end was then purified by ethanol precipitation, lyophilized, and stored at −20 °C under dark conditions.…”

Glycoconjugates: Synthesis, Functional Studies, and Therapeutic Developments

“…Among the GAGs tested, conjugates containing CH and HPN provided the best-balanced profile of in vitro activity and circulation period in mice, suggesting that conjugation with GAGs is a promising strategy for improving the duration of peptide drugs. 237,238 3.2.1.3. Glycosaminoglycan−Nanoparticle Conjugates for Targeted Delivery.…”

“…As an alternative to PEG, highly hydrophilic and biodegradable GAGs such as hyaluronan and heparosan (HPN) have been explored to improve the pharmacokinetics of protein or peptide drugs (Scheme ). Recently, GAG conjugates of two antidiabetic peptide drugs, glucagon-like peptide-1 (GLP-1) and insulin, were shown to have much better half-life and blood-glucose lowering efficacy than unconjugated peptides after subcutaneous injection in mice. , Various GAGs including chondroitin (CH) and heparosan were conjugated to GLP-1 (at engineered Cys35) and insulin (at GlyA1, LysB29, or both) using a hydroxy succinimide-maleimide heterobifunctional linker and various arm lengths. Among the GAGs tested, conjugates containing CH and HPN provided the best-balanced profile of in vitro activity and circulation period in mice, suggesting that conjugation with GAGs is a promising strategy for improving the duration of peptide drugs. , …”

Conjugates for use in peptide therapeutics: A systematic review and meta-analysis

“…Details of all 16 studies are given in the supplementary material (S2 Table ), while the studies that included comparable controls were further analysed as detailed in the next section. Some of the peptides captured in the group of 16 were well-studied native peptides like insulin and glucagon-like peptide 1 (GLP-1) [32][33][34][35][36]. Others were novel, heterologous peptides like PN-2921, an interleukin-6 binding peptide derived from a helix-loop-helix scaffold identified through phage display [37].…”