Selective Agonists and Antagonists of Formylpeptide Receptors: Duplex Flow Cytometry and Mixture-based Positional Scanning Libraries

Authors

Clemencia Pinilla, Torrey Pines Institute for Molecular Studies
Bruce S. Edwards, University of New Mexico
Jon R. Appel, Torrey Pines Institute for Molecular Studies
Tina Yates-Gibbins, Torrey Pines Institute for Molecular Studies
Marc Giulianotti, Torrey Pines Institute for Molecular Studies
Jose L. Medina-Franco, Torrey Pines Institute for Molecular Studies
Susan M. Young, University of New Mexico
Radleigh Santos, Torrey Pines Institute for Molecular StudiesFollow
Larry A. Sklar, University of New Mexico
Richard A. Houghten, Torrey Pines Institute for Molecular Studies

Document Type

Article

Publication Date

9-2013

Publication Title

Molecular Pharmacology

Keywords

Combinatorial chemistry, Receptor binding studies

ISSN

0026-895X

Volume

84

Issue/No.

3

First Page

314

Last Page

324

Abstract

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. Although 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 physiologic roles played by these receptors. High-throughput assays using 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. Screening 37 different mixture-based combinatorial libraries totaling more than five million small molecules (contained in 5,261 mixture samples) resulted in seven libraries that 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 an 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.

Comments

Copyright © 2013 by The American Society for Pharmacology and Experimental Therapeutics

NSUWorks Citation

Pinilla, Clemencia; Edwards, Bruce S.; Appel, Jon R.; Yates-Gibbins, Tina; Giulianotti, Marc; Medina-Franco, Jose L.; Young, Susan M.; Santos, Radleigh; Sklar, Larry A.; and Houghten, Richard A., "Selective Agonists and Antagonists of Formylpeptide Receptors: Duplex Flow Cytometry and Mixture-based Positional Scanning Libraries" (2013). Mathematics Faculty Articles. 228.
https://nsuworks.nova.edu/math_facarticles/228

DOI

10.1124/mol.113.086595