title: Integration of virtual screening with high-throughput flow cytometry to identify novel small molecule formylpeptide receptor antagonists.
abstract: The formylpeptide receptor (FPR) family of G-protein-coupled receptors contributes to the localization and activation of tissue-damaging leukocytes at sites of chronic inflammation. We developed a FPR homology model and pharmacophore (based on the bovine rhodopsin crystal structure and known FPR ligands, respectively) for in silico screening of approximately 480,000 drug-like small molecules. A subset of 4324 compounds that matched the pharmacophore was then physically screened with the HyperCyt flow cytometry platform in high-throughput, no-wash assays that directly measure human FPR binding, with samples (each approximately 2500 cells in 2 microl) analyzed at 40/min. From 52 confirmed hits (1.2% hit rate), we identified 30 potential lead compounds (inhibition constant, Ki= 1-32 microM) representing nine distinct chemical families. Four compounds in one family were weak partial agonists. All others were antagonists. This virtual screening approach improved the physical screening hit rate by 12-fold (versus 0.1% hit-rate in a random compound collection), providing an efficient process for identifying small molecule antagonists.
authors.affiliation: Department of Cytometry, University of New Mexico, Albuquerque, NM 87131-0001, USA.
authors.forename: Bruce S
authors.initials: BS
authors.lastname: Edwards
authors.forename: Cristian
authors.initials: C
authors.lastname: Bologa
authors.forename: Susan M
authors.initials: SM
authors.lastname: Young
authors.forename: Konstantin V
authors.initials: KV
authors.lastname: Balakin
authors.forename: Eric R
authors.initials: ER
authors.lastname: Prossnitz
authors.forename: Nikolay P
authors.initials: NP
authors.lastname: Savchuck
authors.forename: Larry A
authors.initials: LA
authors.lastname: Sklar
authors.forename: Tudor I
authors.initials: TI
authors.lastname: Oprea
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004353:Drug Evaluation, Preclinical; D005434:Flow Cytometry / Q000379:methods*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D044042:Receptors, Formyl Peptide / Q000819:agonists / Q000037:antagonists & inhibitors* / Q000737:chemistry
chemical_list: D044042:Receptors, Formyl Peptide