Peptide Half-Life Extension: Divalent, Small-Molecule Albumin Interactions Direct the Systemic Properties of Glucagon-Like Peptide 1 (GLP-1) Analogues

Abstract

Noncovalent binding of biopharmaceuticals to human serum albumin protects against enzymatic degradation and renal clearance. Herein, we investigated the effect of mono- or divalent small-molecule albumin binders for half-life extension of peptides. For proof-of-principle, the clinically relevant glucagon-like peptide 1 (GLP-1) was functionalized with diflunisal, indomethacin, or both. In vitro, all GLP-1 analogues had subnanomolar GLP-1 receptor potency. Surface plasmon resonance revealed that both small molecules were able to confer albumin affinity to GLP-1 and indicated that affinity is increased for divalent analogues. In lean mice, the divalent GLP-1 analogues were superior to monovalent analogues with respect to control of glucose homeostasis and suppression of food intake. Importantly, divalent GLP-1 analogues showed efficacy comparable to liraglutide, an antidiabetic GLP-1 analogue that carries a long-chain fatty acid. Finally, pharmacokinetic investigations of a divalent GLP-1 analogue demonstrated a promising gain in circulatory half-life and absorption time compared to its monovalent equivalent.

Molecular design opportunities presented by solvent‐exposed regions of target proteins

“…Especially in vivo noncovalent endogenous HSA targeting has received widespread attention. For example, the clinically relevant glucagon‐like peptide 1 (GLP‐1) was linked with diflunisal ( 196 , albumin binder) and indomethacin ( 197 , albumin binder) to generate a divalent GLP‐1 derivative 198 with a big boost in circulatory half‐life and absorption time relative to its monovalent equivalent in PK studies (Figure ) . Again, the key was finding a suitable tethering site for the divalent HAS‐conjugate.…”

“…Furthermore, 174 shows low nanomolar activity, affording 4.3-fold and 4300-fold improvement of polymerization inhibition in vitro compared with the parent TMC derivative and d4T, respectively. 147 The bifunctional chimeric HIV inhibitor 177 was obtained by covalently combining a small-molecular CCR5 antagonist, LGC240 (176), and a gp120 peptide triazole antagonist, UM15 (175) (Figure 25). Interestingly, 177…”

“…For example, the clinically relevant glucagon-like peptide 1 (GLP-1) was linked with diflunisal (196, albumin binder) and indomethacin (197, albumin binder) to generate a divalent GLP-1 derivative 198 with a big boost in circulatory half-life and absorption time relative to its monovalent equivalent in PK studies ( Figure 29). 175 Again, the key was finding a suitable tethering site for the divalent HAS-conjugate. Similar insights have proved useful in the structure-based exploitation of other drug delivery systems, including human serum amyloid P component, polyamines, and antibody-drug conjugates (including ARMs).…”

Discovery of lixisenatide analogues as long-acting hypoglycemic agents using novel peptide half-life extension technology based on mycophenolic acid

“…Thus, alternative strategies for mediating serum albumin interactions with enhanced solubility, HSA affinity or additional therapeutic property continues to be attractive. 24,25 Herein, we explored an alternative methodology to conjugate of novel small molecule albumin binders to peptides. Some anionic aromatic small molecules have high albumin binding affinities and can be used as an alternative to lipidation.…”

The chronic administration of two novel long‐acting Xenopus glucagon‐like peptide‐1 analogs xGLP159 and XGLP296 potently improved systemic metabolism and glycemic control in rodent models

“…The main obstacle for the therapeutic use of GLP-1 is its short circulating half-life (t 1/2 ) (7). Hence, considerable research interests have focused on the development of degradation-resistant long-acting GLP-1 analogs (8,9). To date, 6 GLP-1 receptor agonists in Europe and the United States have been approved for treating type 2 diabetes mellitus, and many others are in clinical development.…”