Expression and Characterization of a Potent Long-Acting GLP-1 Receptor Agonist, GLP-1-IgG2σ-Fc

PLoS One. 2016 May 27;11(5):e0156449. doi: 10.1371/journal.pone.0156449. eCollection 2016.

Abstract

Human GLP-1 (glucagon-like peptide-1) can produce a remarkable improvement in glycemic control in patients with type 2 diabetes. However, its clinical benefits are limited by its short half-life, which is less than 2 min because of its small size and rapid enzymatic inactivation by dipeptidyl peptidase IV. We engineered GLP-1-IgG2σ-Fc, a 68-kDa fusion protein linking a variant human GLP-1 (A8G/G26E/R36G) to a human IgG2σ constant heavy-chain. A stably transfected Chinese hamster ovary cell line was obtained using electroporation. Western blotting showed that the expressed protein was immunoreactive to both GLP-1 and IgG antibodies. GLP-1-IgG2σ-Fc stimulated insulin secretion from INS-1 cells in a dose- and glucose-dependent manner and increased insulin mRNA expression. The half-life of GLP-1-IgG2σ-Fc in cynomolgus monkeys was approximately 57.1 ± 4.5 h. In the KKAy mouse model of diabetes, one intraperitoneal injection of GLP-1-IgG2σ-Fc (1 mg/kg) reduced blood glucose levels for 5 days. A 4-week repeat-administration study identified sustained effects on blood glucose levels. Oral glucose tolerance tests conducted at the beginning and end of this 4-week period showed that GLP-1-IgG2σ-Fc produced a stable glucose lowering effect. In addition, KKAy mice treated with GLP-1-IgG2σ-Fc showed statistically significant weight loss from day 23. In conclusion, these properties of GLP-1-IgG2σ-Fc demonstrated that it represented a potential long-acting GLP-1 receptor agonist for the treatment of type 2 diabetes.

MeSH terms

  • Animals
  • Body Weight / drug effects
  • Cell Line
  • Gene Expression
  • Glucagon-Like Peptide 1 / genetics*
  • Glucagon-Like Peptide 1 / isolation & purification
  • Glucagon-Like Peptide 1 / pharmacokinetics
  • Glucagon-Like Peptide 1 / pharmacology*
  • Glucagon-Like Peptide-1 Receptor / agonists*
  • Glucose / metabolism
  • Humans
  • Immunoglobulin G / genetics
  • Macaca fascicularis
  • Male
  • Mice
  • Postprandial Period / drug effects
  • Protein Engineering*
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / pharmacokinetics
  • Recombinant Fusion Proteins / pharmacology*

Substances

  • Glucagon-Like Peptide-1 Receptor
  • Immunoglobulin G
  • Recombinant Fusion Proteins
  • Glucagon-Like Peptide 1
  • Glucose

Grants and funding

This work was supported by the Natural Science Foundation of China (Haifeng Song; 81272701). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.