title: Discovery and Characterization of Novel GPR39 Agonists Allosterically Modulated by Zinc.
abstract: In this study, we identified two previously described kinase inhibitors-3-(4-chloro-2-fluorobenzyl)-2-methyl-N-(3-methyl-1H-pyrazol-5-yl)-8-(morpholinomethyl)imidazo[1,2-b]pyridazin-6-amine (LY2784544) and 1H-benzimidazole-4-carboxylic acid, 2-methyl-1-[[2-methyl-3-(trifluoromethyl)phenyl]methyl]-6-(4-morpholinyl)- (GSK2636771)-as novel GPR39 agonists by unbiased small-molecule-based screening using a β-arrestin recruitment screening approach (PRESTO-Tango). We characterized the signaling of LY2784544 and GSK2636771 and compared their signaling patterns with a previously described "GPR39-selective" agonist N-[3-chloro-4-[[[2-(methylamino)-6-(2-pyridinyl)-4- pyrimidinyl]amino]methyl]phenyl]methanesulfonamide (GPR39-C3) at both canonical and noncanonical signaling pathways. Unexpectedly, all three compounds displayed probe-dependent and pathway-dependent allosteric modulation by concentrations of zinc reported to be physiologic. LY2784544 and GS2636771 at GPR39 in the presence of zinc were generally as potent or more potent than their reported activities against kinases in whole-cell assays. These findings reveal an unexpected role of zinc as an allosteric potentiator of small-molecule-induced activation of GPR39 and expand the list of potential kinase off-targets to include understudied G protein-coupled receptors.
authors.affiliation: Department of Pharmacology (S.S., X.-P.H., W.K.K., B.L.R.) and National Institute of Mental Health Psychoactive Drug Screening Program (X.-P.H., B.L.R.), School of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Seiji
authors.initials: S
authors.lastname: Sato
authors.affiliation: Department of Pharmacology (S.S., X.-P.H., W.K.K., B.L.R.) and National Institute of Mental Health Psychoactive Drug Screening Program (X.-P.H., B.L.R.), School of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Xi-Ping
authors.initials: XP
authors.lastname: Huang
authors.affiliation: Department of Pharmacology (S.S., X.-P.H., W.K.K., B.L.R.) and National Institute of Mental Health Psychoactive Drug Screening Program (X.-P.H., B.L.R.), School of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Wesley K
authors.initials: WK
authors.lastname: Kroeze
authors.affiliation: Department of Pharmacology (S.S., X.-P.H., W.K.K., B.L.R.) and National Institute of Mental Health Psychoactive Drug Screening Program (X.-P.H., B.L.R.), School of Medicine, University of North Carolina, Chapel Hill, North Carolina bryan_roth@med.unc.edu.
authors.forename: Bryan L
authors.initials: BL
authors.lastname: Roth
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D055808:Drug Discovery; D004353:Drug Evaluation, Preclinical; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000378:metabolism; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism; D006639:Histidine / Q000235:genetics; D006801:Humans; D006868:Hydrolysis; D009154:Mutation / Q000235:genetics; D010716:Phosphatidylinositols / Q000378:metabolism; D011743:Pyrimidines / Q000494:pharmacology; D043562:Receptors, G-Protein-Coupled / Q000819:agonists* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013449:Sulfonamides / Q000494:pharmacology; D015032:Zinc / Q000494:pharmacology*; D000071557:beta-Arrestins / Q000378:metabolism
chemical_list: C000621962:GPR39-C3; D010716:Phosphatidylinositols; D011743:Pyrimidines; D043562:Receptors, G-Protein-Coupled; D013449:Sulfonamides; D000071557:beta-Arrestins; D006639:Histidine; D000242:Cyclic AMP; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D019205:GTP-Binding Protein alpha Subunits, Gs; D015032:Zinc