Development of antagonist molecular probes of the P2Y14 G protein-coupled receptor

Thursday, November 07, 2013 — Poster Session II
12:00 p.m. – 2:00 p.m.	FAES Academic Center (Upper-Level Terrace)	NIDDK	PHARM-7

Authors

E.A. Kiselev
S. Paoletta
M. Barrett
T.K. Harden
K.A. Jacobson

Abstract

The P2Y14 receptor (P2Y14R) is a member of the P2Y GPCR family. UDP and UDP-glucose act as endogenous agonists of this Gi protein-coupled receptor to inhibit cAMP production. P2Y14R mediates inflammation and hypoxic stress in numerous tissues, making it an attractive object for biological and pharmacological evaluation. Studies with P2Y14R knockout mice have demonstrated its value as a potential drug target for treatment of diabetes. We strive toward the goal of introducing methods for fluorescent characterization of the P2Y14R, which will satisfy the need for affinity probes. A novel high-throughput screening method of assessing affinities of P2Y14R ligands will be based on newly synthesized probes for fluorescence polarization or flow cytometry measurements. The template antagonist in question, 4-((piperidin-4-yl)-phenyl)-(7-(4-(trifluoromethyl)-phenyl)-2-naphthoic acid, is shown to be selective for P2Y14R with a Ki of 0.5 nM. Two homology models (A2A- and CXCR4-based) have been constructed and used in ligand design through virtual docking. The derivatives of PPTN have been furnished with alkynyl or amino groups, suitable for attachment of fluorescent moieties by Click chemistry or amide coupling. Antagonist activities of synthetic analogues are evaluated in C6 rat glioma cells stably expressing human P2Y14R. Preliminary results suggest that P2Y14R potency is preserved by chain functionalization.