title: Pharmacological regulation of insulin secretion in MIN6 cells through the fatty acid receptor GPR40: identification of agonist and antagonist small molecules.
abstract: 1. Long chain fatty acids have recently been identified as agonists for the G protein-coupled receptors GPR40 and GPR120. Here, we present the first description of GW9508, a small-molecule agonist of the fatty acid receptors GPR40 and GPR120. In addition, we also describe the pharmacology of GW1100, a selective GPR40 antagonist. These molecules were used to further investigate the role of GPR40 in glucose-stimulated insulin secretion in the MIN6 mouse pancreatic beta-cell line. 2. GW9508 and linoleic acid both stimulated intracellular Ca2+ mobilization in human embryonic kidney (HEK)293 cells expressing GPR40 (pEC50 values of 7.32+/-0.03 and 5.65+/-0.06, respectively) or GPR120 (pEC50 values of 5.46+/-0.09 and 5.89+/-0.04, respectively), but not in the parent HEK-293 cell line. 3. GW1100 dose dependently inhibited GPR40-mediated Ca2+ elevations stimulated by GW9508 and linoleic acid (pIC50 values of 5.99+/-0.03 and 5.99+/-0.06, respectively). GW1100 had no effect on the GPR120-mediated stimulation of intracellular Ca2+ release produced by either GW9508 or linoleic acid. 4. GW9508 dose dependently potentiated glucose-stimulated insulin secretion in MIN6 cells, but not in primary rat or mouse islets. Furthermore, GW9508 was able to potentiate the KCl-mediated increase in insulin secretion in MIN6 cells. The effects of GW9508 on insulin secretion were reversed by GW1100, while linoleic acid-stimulated insulin secretion was partially attenuated by GW1100. 5. These results add further evidence to a link between GPR40 and the ability of fatty acids to acutely potentiate insulin secretion and demonstrate that small-molecule GPR40 agonists are glucose-sensitive insulin secretagogues.
authors.affiliation: Department of Metabolic Diseases, GlaxoSmithKline, 5 Moore Drive, Research Triangle Park, NC 27709, USA. cpb15172@gsk.com
authors.forename: Celia P
authors.initials: CP
authors.lastname: Briscoe
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Peat
authors.forename: Stephen C
authors.initials: SC
authors.lastname: McKeown
authors.forename: David F
authors.initials: DF
authors.lastname: Corbett
authors.forename: Aaron S
authors.initials: AS
authors.lastname: Goetz
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Littleton
authors.forename: David C
authors.initials: DC
authors.lastname: McCoy
authors.forename: Terry P
authors.initials: TP
authors.lastname: Kenakin
authors.forename: John L
authors.initials: JL
authors.lastname: Andrews
authors.forename: Carina
authors.initials: C
authors.lastname: Ammala
authors.forename: James A
authors.initials: JA
authors.lastname: Fornwald
authors.forename: Diane M
authors.initials: DM
authors.lastname: Ignar
authors.forename: Stephen
authors.initials: S
authors.lastname: Jenkinson
mesh_terms: D000818:Animals; D001565:Benzoates / Q000494:pharmacology; D016466:CHO Cells; D002460:Cell Line; D002478:Cells, Cultured; D006224:Cricetinae; D004353:Drug Evaluation, Preclinical; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology; D000960:Hypolipidemic Agents / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism*; D008744:Methylamines / Q000494:pharmacology; D051379:Mice; D008954:Models, Biological; D011189:Potassium Chloride / Q000494:pharmacology; D011422:Propionates / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D043562:Receptors, G-Protein-Coupled / Q000819:agonists* / Q000037:antagonists & inhibitors*; D018411:Spodoptera / Q000166:cytology
chemical_list: D001565:Benzoates; C473034:FFAR1 protein, human; C498007:FFAR4 protein, mouse; C515631:GW1100; C515630:GW9508; D007004:Hypoglycemic Agents; D000960:Hypolipidemic Agents; D007328:Insulin; D008744:Methylamines; D011422:Propionates; D011743:Pyrimidines; D043562:Receptors, G-Protein-Coupled; D011189:Potassium Chloride; D005947:Glucose