Glucagon from the phylogenetically ancient paddlefish provides a template for the design of a long-acting peptide with effective anti-diabetic and anti-obesity activities

This study has examined the in vitro and in vivo anti-diabetic properties of the peptidase-resistant analogues [D-Ser(2)]palmitoyl-paddlefish glucagon and [D-Ser(2)]palmitoyl-lamprey glucagon. The peptides stimulated insulin release from BRIN-BD11 clonal β-cells and isolated mouse pancreatic islets and also enhanced cAMP production in cells transfected with the human GLP-1 receptor and with the human glucagon receptor. The insulinotropic actions of the peptides were attenuated in INS-1 cells lacking GLP-1 and glucagon receptors. [D-Ser(2)]palmitoyl-paddlefish glucagon stimulated proliferation of BRIN-BD11 cells and protected against cytokine-mediated apoptosis as effectively as GLP-1. The analogue was more effective than the native peptide or the lamprey glucagon analogue in acutely lowering blood glucose and elevating plasma insulin in lean mice even when administered up to 4 h before a glucose load. Twice daily administration of [D-Ser(2)]palmitoyl-paddlefish glucagon to high-fat fed mice over 21 days reduced food intake, body weight, non-fasting blood glucose and plasma insulin concentrations, as well as significantly improving glucose tolerance and insulin resistance and decreasing α-cell area and pancreatic insulin content. Islet expression of the Gcgr, Glp1r, Gipr and Slc2a2 (GLUT-2) genes significantly increased. These data demonstrate that long-acting peptide [D-Ser(2)]palmitoyl-paddlefish glucagon exerts beneficial metabolic properties in diabetic mice via Ggcr- and Glp1r-activated pathways and so shows potential as a template for further development into an agent for treatment of patients with obesity-related Type 2 diabetes.