title: A high-throughput small-molecule ligand screen targeted to agonists and antagonists of the G-protein-coupled receptor GPR54.
abstract: Recent data have shown that the G-protein-coupled receptor GPR54 (also known as KiSS-1 receptor) regulates GnRH release from the hypothalamus. This essential role of GPR54 in controlling the hypothalamic-pituitary-gonadal axis makes it an attractive target for therapeutic intervention in reproductive and cancer medicine. Currently, there are no small-molecule modulators of GPR54 function for experimental or clinical use. To identify small-molecule compounds that modify GPR54 signal transduction, the authors have adapted a cell-based functional assay for high-throughput screening (HTS) using a commercially available homogeneous time-resolved fluorescence assay for inositol phosphate accumulation. They generated stable Chinese hamster ovary cell transfectants that express human GPR54 for use in this assay. After optimization in an automated HTS environment, they screened a library of 110,000 small-molecule compounds using 2 protocols, one to identify agonists and one to identify antagonists. Hits obtained in the primary screen were confirmed to be active in secondary in vitro assays. Compounds identified as agonists or antagonists from HTS and secondary screening will be characterized to identify agents with the potential to be developed as novel orally active agents to treat hormone-dependent disorders such as abnormal puberty, infertility, endometriosis, and sex steroid-dependent tumors.
authors.affiliation: Department of Obstetrics and Gynecology, Boston University School of Medicine, USA.
authors.forename: Wendy
authors.initials: W
authors.lastname: Kuohung
authors.forename: Maria
authors.initials: M
authors.lastname: Burnett
authors.forename: Deepa
authors.initials: D
authors.lastname: Mukhtyar
authors.forename: Eli
authors.initials: E
authors.lastname: Schuman
authors.forename: Jake
authors.initials: J
authors.lastname: Ni
authors.forename: William F
authors.initials: WF
authors.lastname: Crowley
authors.forename: Marcie A
authors.initials: MA
authors.lastname: Glicksman
authors.forename: Ursula B
authors.initials: UB
authors.lastname: Kaiser
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D008024:Ligands; D043562:Receptors, G-Protein-Coupled / Q000819:agonists* / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000074502:Receptors, Kisspeptin-1
chemical_list: D007295:Inositol Phosphates; C118688:KISS1R protein, human; D008024:Ligands; D043562:Receptors, G-Protein-Coupled; D000074502:Receptors, Kisspeptin-1; D048049:Extracellular Signal-Regulated MAP Kinases
references.citation: J Biomol Screen. 1999;4(2):67-73
references.pmid: 10838414
references.citation: N Engl J Med. 2003 Oct 23;349(17):1614-27
references.pmid: 14573733
references.citation: Biochem Biophys Res Commun. 2003 Dec 26;312(4):1357-63
references.pmid: 14652023
references.citation: Cancer Res. 2005 Nov 15;65(22):10450-6
references.pmid: 16288036
references.citation: Biochem Biophys Res Commun. 2004 Jul 23;320(2):383-8
references.pmid: 15219839
references.citation: Bioorg Med Chem Lett. 2006 Jan 1;16(1):134-7
references.pmid: 16242330
references.citation: Mol Pharmacol. 2009 Jul;76(1):58-67
references.pmid: 19389922
references.citation: Endocrinology. 2005 Jul;146(7):2976-84
references.pmid: 15831567
references.citation: Proc Natl Acad Sci U S A. 2005 Feb 8;102(6):2129-34
references.pmid: 15684075
references.citation: J Biomol Screen. 2007 Mar;12(2):285-7
references.pmid: 17289936
references.citation: Annu Rev Physiol. 2008;70:213-38
references.pmid: 17988212
references.citation: Endocrinology. 2004 Sep;145(9):4073-7
references.pmid: 15217982
references.citation: Int J Cancer. 1997 Jun 11;71(6):1035-44
references.pmid: 9185708
references.citation: J Clin Endocrinol Metab. 2005 Dec;90(12):6609-15
references.pmid: 16174713
references.citation: Nature. 2001 May 31;411(6837):613-7
references.pmid: 11385580
references.citation: Endocrinology. 2006 May;147(5):2122-6
references.pmid: 16469799
references.citation: J Neurosci. 2006 Jun 21;26(25):6687-94
references.pmid: 16793876
references.citation: Proc Natl Acad Sci U S A. 2003 Sep 16;100(19):10972-6
references.pmid: 12944565
references.citation: J Neurosci. 2005 Dec 7;25(49):11349-56
references.pmid: 16339030
references.citation: J Med Chem. 2007 Feb 8;50(3):462-71
references.pmid: 17266198
references.citation: Adv Drug Deliv Rev. 2001 Mar 1;46(1-3):3-26
references.pmid: 11259830
references.citation: Neurosci Lett. 2006 Jul 3;401(3):225-30
references.pmid: 16621281
references.citation: Biochem Biophys Res Commun. 2005 Jan 21;326(3):677-86
references.pmid: 15596153
references.citation: Endocrinology. 2005 Sep;146(9):3686-92
references.pmid: 15919741
references.citation: Endocrinology. 2004 Oct;145(10):4565-74
references.pmid: 15242985
references.citation: J Neurosci. 2009 Mar 25;29(12):3920-9
references.pmid: 19321788
references.citation: J Clin Endocrinol Metab. 2003 Feb;88(2):834-43
references.pmid: 12574221
references.citation: Endocrinology. 2005 Jan;146(1):156-63
references.pmid: 15375028
references.citation: J Natl Cancer Inst. 1996 Dec 4;88(23):1731-7
references.pmid: 8944003
references.citation: Biochem Biophys Res Commun. 2001 Sep 7;286(5):958-63
references.pmid: 11527393