New screening strategy and analysis for identification of allosteric modulators for glucagon-like peptide-1 receptor using GLP-1 (9-36) amide.
The glucagon-like peptide-1 receptor (GLP-1R) is an important physiologic regulator of insulin secretion and a major therapeutic target for diabetes mellitus. GLP-1 (7-36) amide (active form of GLP-1) is truncated to GLP-1 (9-36) amide, which has been described as a weak agonist of GLP-1R and the major form of GLP-1 in the circulation. New classes of positive allosteric modulators (PAMs) for GLP-1R may offer improved therapeutic profiles. To identify these new classes, we developed novel and robust primary and secondary high-throughput screening (HTS) systems in which PAMs were identified to enhance the GLP-1R signaling induced by GLP-1 (9-36) amide. Screening enabled identification of two compounds, HIT-465 and HIT-736, which possessed new patterns of modulation of GLP-1R. We investigated the ability of these compounds to modify GLP-1R signaling enhanced GLP-1 (9-36) amide- and/or GLP-1 (7-36) amide-mediated cyclic adenosine monophosphate (cAMP) accumulation. These compounds also had unique profiles with regard to allosteric modulation of multiple downstream signaling (PathHunter β-arrestin signaling, PathHunter internalization signaling, microscopy-based internalization assay). We found allosteric modulation patterns to be obviously different among HIT-465, HIT-736, and Novo Nordisk compound 2. This work may enable the design of new classes of drug candidates by targeting modulation of GLP-1 (7-36) amide and GLP-1 (9-36) amide.
Genomic Science Laboratories, Sumitomo Dainippon Pharma, Konohana-ku, Osaka 554-0022, Japan.
Atsushi
A
Nakane
Genomic Science Laboratories, Sumitomo Dainippon Pharma, Konohana-ku, Osaka 554-0022, Japan.
Yusuke
Y
Gotoh
Genomic Science Laboratories, Sumitomo Dainippon Pharma, Konohana-ku, Osaka 554-0022, Japan.
Junji
J
Ichihara
Genomic Science Laboratories, Sumitomo Dainippon Pharma, Konohana-ku, Osaka 554-0022, Japan. Electronic address: hidetaka-nagata@ds-pharma.co.jp.
Hidetaka
H
Nagata
D000494:Allosteric Regulation / Q000187:drug effects; D000577:Amides / Q000737:chemistry*; D000818:Animals; D019390:Arrestins / Q000378:metabolism; D016466:CHO Cells; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D000242:Cyclic AMP / Q000378:metabolism; D003920:Diabetes Mellitus / Q000188:drug therapy; D052216:Glucagon-Like Peptide 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000067757:Glucagon-Like Peptide-1 Receptor / Q000235:genetics / Q000378:metabolism*; D057166:High-Throughput Screening Assays; D006801:Humans; D007004:Hypoglycemic Agents / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use; D016297:Mutagenesis, Site-Directed; D015398:Signal Transduction / Q000187:drug effects; D000071557:beta-Arrestins; D000097789:Glucagon-Like Peptide-1 Receptor Agonists
D000577:Amides; D019390:Arrestins; D000067757:Glucagon-Like Peptide-1 Receptor; D007004:Hypoglycemic Agents; D000071557:beta-Arrestins; D052216:Glucagon-Like Peptide 1; D000242:Cyclic AMP; D002118:Calcium; D000097789:Glucagon-Like Peptide-1 Receptor Agonists
G-protein-coupled receptor (GPCR); Glucagon-like peptide-1 (GLP-1); High-throughput screening (HTS); Positive allosteric modulator (PAM); Receptor internalization; β-arrestin