PT  - JOURNAL ARTICLE
AU  - Edwards, Bruce S.
AU  - Bologa, Cristian
AU  - Young, Susan M.
AU  - Balakin, Konstantin V.
AU  - Prossnitz, Eric R.
AU  - Savchuck, Nikolay P.
AU  - Sklar, Larry A.
AU  - Oprea, Tudor I.
TI  - Integration of Virtual Screening with High-Throughput Flow Cytometry to Identify Novel Small Molecule Formylpeptide Receptor Antagonists
AID  - 10.1124/mol.105.014068
DP  - 2005 Nov 01
TA  - Molecular Pharmacology
PG  - 1301--1310
VI  - 68
IP  - 5
4099  - http://molpharm.aspetjournals.org/content/68/5/1301.short
4100  - http://molpharm.aspetjournals.org/content/68/5/1301.full
SO  - Mol Pharmacol2005 Nov 01; 68
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.