RT Journal Article
SR Electronic
T1 Integration of Virtual Screening with High-Throughput Flow Cytometry to Identify Novel Small Molecule Formylpeptide Receptor Antagonists
JF Molecular Pharmacology
JO Mol Pharmacol
FD American Society for Pharmacology and Experimental Therapeutics
SP 1301
OP 1310
DO 10.1124/mol.105.014068
VO 68
IS 5
A1 Bruce S. Edwards
A1 Cristian Bologa
A1 Susan M. Young
A1 Konstantin V. Balakin
A1 Eric R. Prossnitz
A1 Nikolay P. Savchuck
A1 Larry A. Sklar
A1 Tudor I. Oprea
YR 2005
UL http://molpharm.aspetjournals.org/content/68/5/1301.abstract
AB 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 ∼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 ∼2500 cells in 2 μl) analyzed at 40/min. From 52 confirmed hits (1.2% hit rate), we identified 30 potential lead compounds (inhibition constant, Ki = 1-32 μM) 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.