TY - JOUR T1 - Integration of Virtual Screening with High-Throughput Flow Cytometry to Identify Novel Small Molecule Formylpeptide Receptor Antagonists JF - Molecular Pharmacology JO - Mol Pharmacol SP - 1301 LP - 1310 DO - 10.1124/mol.105.014068 VL - 68 IS - 5 AU - Bruce S. Edwards AU - Cristian Bologa AU - Susan M. Young AU - Konstantin V. Balakin AU - Eric R. Prossnitz AU - Nikolay P. Savchuck AU - Larry A. Sklar AU - Tudor I. Oprea Y1 - 2005/11/01 UR - http://molpharm.aspetjournals.org/content/68/5/1301.abstract N2 - 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. ER -