Polypharmacy through Phage Display: Selection of Glucagon and GLP-1 Receptor Co-agonists from a Phage-Displayed Peptide Library

Abstract

A promising emerging area for the treatment of obesity and diabetes is combinatorial hormone therapy, where single-molecule peptides are rationally designed to integrate the complementary actions of multiple endogenous metabolically-related hormones. We describe here a proof-of-concept study on developing unimolecular polypharmacy agents through the use of selection methods based on phage-displayed peptide libraries (PDL). Co-agonists of the glucagon (GCG) and GLP-1 receptors were identified from a PDL sequentially selected on GCGR- and GLP1R-overexpressing cells. After two or three rounds of selection, 7.5% of randomly picked clones were GLP1R/GCGR co-agonists, and a further 1.53% were agonists of a single receptor. The phages were sequenced and 35 corresponding peptides were synthesized. 18 peptides were potent co-agonists, 8 of whom showed EC50 ≤ 30 pM on each receptor, comparable to the best rationally designed co-agonists reported in the literature. Based on literature examples, two sequences were engineered to stabilize against dipeptidyl peptidase IV cleavage and prolong the in vivo half-life: the engineered peptides were comparably potent to the parent peptides on both receptors, highlighting the potential use of phage-derived peptides as therapeutic agents. The strategy described here appears of general value for the discovery of optimized polypharmacology paradigms across several metabolically-related hormones.

Type 2 Diabetes Mellitus Associated with Obesity (Diabesity). The Central Role of Gut Microbiota and Its Translational Applications

“…In addition, the use of phage-derived products poses an interesting area of research in the clinical management of diabesity [ 236 ]. The childhood could be an interesting period to target gut microbiota, either using probiotics, prebiotics, phages or a combination of these techniques [ 237 ], and clearly strengthening a proper lifestyle. However, currently, there are no clinical trials using phage therapy in patients with diabesity.…”

Peptide Conjugates with Small Molecules Designed to Enhance Efficacy and Safety

“…Similar to drug discovery directed at small molecules, peptide research has evolved in the direction of multimode pharmacology, [20,21,22] where single molecules activate multiple receptors in an additive and occasionally in a synergistic manner to achieve superior efficacy often at reduced dose [1,2,3,4,23,24,25]. This type of pharmacology is exemplified in purposefully integrated, dual agonism at amylin and calcitonin, GLP-1 and glucagon, or with gastric inhibitory peptide (GIP), and triple agonism at GLP-1, glucagon and GIP in treatment of the metabolic syndrome [26,27,28,29,30,31,32,33,34,35]. The sequence of these multi-action peptides largely derives from intermixing resides from each native hormone to achieve balanced, full agonism at the respective cognate receptors.…”

High-dimensionality Data Analysis of Pharmacological Systems Associated with Complex Diseases

“…The identification of the hypothalamus as a potential master controller in this process then helps target drug receptor and neurotransmitter signaling systems that may engender true systemic therapeutic effects (Phan et al, 2005;Amri and Pisani, 2016;Scarpace et al, 2016;Steculorum et al, 2017;Kim and Choe, 2019;Mravec et al, 2019). The major current hindrance to the generation of rationally designed network level therapeutics that can control such a complex system is the need to inculcate the concept of effectively engineered polypharmacology as a new route for drug development and discovery (Demartis et al, 2018;Grisoni et al, 2019;Peón et al, 2019). Given this, it will also be vital to refine the drug refinement and approval process (FDA and European Medicines Agency) to acknowledge that with an H-D appreciation of both disease drug response activities comes a need to accept molecular complexity when approving new agents for complex disorders.…”