A vitamin B12 conjugate of exendin-4 improves glucose tolerance without associated nausea or hypophagia in rodents

Diabetes Obes Metab. 2018 May;20(5):1223-1234. doi: 10.1111/dom.13222. Epub 2018 Feb 20.

Abstract

Aims: While pharmacological glucagon-like peptide-1 receptor (GLP-1R) agonists are FDA-approved for treating type 2 diabetes mellitus (T2DM) and obesity, a major side effect is nausea/malaise. We recently developed a conjugate of vitamin B12 (B12) bound to the GLP-1R agonist exendin-4 (Ex4), which displays enhanced proteolytic stability and retention of GLP-1R agonism. Here, we evaluate whether the conjugate (B12-Ex4) can improve glucose tolerance without producing anorexia and malaise.

Materials and methods: We evaluated the effects of systemic B12-Ex4 and unconjugated Ex4 on food intake and body weight change, oral glucose tolerance and nausea/malaise in male rats, and on intraperitoneal glucose tolerance in mice. To evaluate whether differences in the profile of effects of B12-Ex4 vs unconjugated Ex4 are the result of altered CNS penetrance, rats received systemic injections of fluorescein-Ex4 (Flex), Cy5-B12 or Cy5-B12-Ex4 and brain penetrance was evaluated using confocal microscopy. Uptake of systemically administered Cy5-B12-Ex4 in insulin-containing pancreatic beta cells was also examined.

Results: B12-Ex4 conjugate improves glucose tolerance, but does not elicit the malaise and anorexia produced by unconjugated Ex4. While Flex robustly penetrates into the brain (dorsal vagal complex, paraventricular hypothalamus), Cy5-B12 and Cy5-B12-Ex4 fluorescence were not observed centrally, supporting an absence of CNS penetrance, in line with observed reduction in CNS-associated Ex4 side effects. Cy5-B12-Ex4 colocalizes with insulin in the pancreas, suggesting direct pancreatic action as a potential mechanism underlying the hypoglycaemic effects of B12-Ex4.

Conclusion: These novel findings highlight the potential clinical utility of B12-Ex4 conjugates as possible future T2DM therapeutics with reduced incidence of adverse effects.

Keywords: antidiabetic drug; appetite control; drug development; exenatide.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Appetite Regulation / drug effects*
  • Behavior, Animal / drug effects
  • Blood-Brain Barrier / drug effects
  • Blood-Brain Barrier / metabolism
  • Drug Stability
  • Energy Intake / drug effects
  • Energy Metabolism / drug effects
  • Exenatide / adverse effects
  • Exenatide / analogs & derivatives*
  • Exenatide / pharmacokinetics
  • Exenatide / therapeutic use
  • Female
  • Glucagon-Like Peptide-1 Receptor / agonists*
  • Glucagon-Like Peptide-1 Receptor / genetics
  • Glucagon-Like Peptide-1 Receptor / metabolism
  • Glucose Intolerance / blood
  • Glucose Intolerance / drug therapy*
  • Glucose Intolerance / metabolism
  • Glucose Intolerance / pathology
  • HEK293 Cells
  • Humans
  • Hypoglycemic Agents / adverse effects
  • Hypoglycemic Agents / chemistry
  • Hypoglycemic Agents / pharmacokinetics
  • Hypoglycemic Agents / therapeutic use*
  • Insulin-Secreting Cells / metabolism
  • Insulin-Secreting Cells / pathology
  • Male
  • Mice, Inbred C57BL
  • Nausea / chemically induced
  • Nausea / prevention & control
  • Rats, Sprague-Dawley
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Tissue Distribution
  • Vitamin B 12 / adverse effects
  • Vitamin B 12 / analogs & derivatives*
  • Vitamin B 12 / pharmacokinetics
  • Vitamin B 12 / therapeutic use

Substances

  • Glucagon-Like Peptide-1 Receptor
  • Hypoglycemic Agents
  • Recombinant Proteins
  • Exenatide
  • Vitamin B 12