RT Journal Article
SR Electronic
T1 Selective Agonists and Antagonists of Formylpeptide Receptors: Duplex Flow Cytometry and Mixture-based Positional Scanning Libraries
JF Molecular Pharmacology
JO Mol Pharmacol
FD American Society for Pharmacology and Experimental Therapeutics
SP mol.113.086595
DO 10.1124/mol.113.086595
A1 Clemencia Pinilla
A1 Bruce S. Edwards
A1 Jon R. Appel
A1 Tina Yates-Gibbins
A1 Marc A. Giulianotti
A1 Jose L. Medina-Franco
A1 Susan M. Young
A1 Radleigh G. Santos
A1 Larry A. Sklar
A1 Richard A. Houghten
YR 2013
UL http://molpharm.aspetjournals.org/content/early/2013/06/20/mol.113.086595.abstract
AB The formylpeptide receptor (FPR1) and formylpeptide-like 1 receptor (FPR2) are G protein coupled receptors that are linked to acute inflammatory responses, malignant glioma stem cell metastasis and chronic inflammation. While several N-formyl peptides are known to bind to these receptors, more selective small molecule high-affinity ligands are needed for a better understanding of the physiological roles played by these receptors. High throughput assays utilizing mixture-based combinatorial libraries represent a unique, highly efficient approach for rapid data acquisition and ligand identification. We report the superiority of this approach in the context of the simultaneous screening of a diverse set of mixture-based small molecule libraries. We used a single cross-reactive peptide ligand for a duplex flow cytometric screen of FPR1 and FPR2 in color-coded cell lines. Upon screening 37 different mixture-based combinatorial libraries totaling more than 5 million small molecules (contained in 5,261 mixture samples), seven libraries significantly inhibited activity at the receptors. Using positional scanning deconvolution, selective high affinity (low nM Ki) individual compounds were identified from two separate libraries, namely pyrrolidine bis-diketopiperazine and polyphenyl urea. The most active individual compounds were characterized for their functional activities as agonists or antagonists with the most potent FPR1 agonist and FPR2 antagonist identified to date with an EC50 of 131 nM (4 nM Ki) and IC50 of 81 nM (1 nM Ki), respectively, in intracellular Ca2+ response determinations. Comparative analyses of other previous screening approaches clearly illustrate the efficiency of identifying receptor selective, individual compounds from mixture-based combinatorial libraries.