A selective high affinity antagonist of the P2Y14 receptor inhibits UDP-glucose-stimulated chemotaxis of human neutrophils

MATTHEW O. BARRETT; JULIANA I. SESMA; CHRISTOPHER B. BALL; P. SURESH JAYASEKARA; KENNETH A. JACOBSON; EDUARDO R. LAZAROWSKI; T. KENDALL HARDEN

MOLECULAR PHARMACOLOGY

AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS

Lugar: Baltimore; Año: 2013 vol. 84 p. 41 - 49

0026-895X

The nucleotide-sugar?activated P2Y14 receptor (P2Y14-R) is highly expressed in hematopoietic cells. Although the physiologic func- tions of this receptor remain undefined, it has been strongly implicated recently in immune and inflammatory responses. Lack of availability of receptor-selective high-affinity antag- onists has impeded progress in studies of this and most of the eight nucleotide-activated P2Y receptors. A series of mole- cules recently were identified by Gauthier et al. (Gauthier et al., 2011) that exhibited antagonist activity at the P2Y14-R. We synthesized one of these molecules, a 4,7-disubstituted 2- naphthoic acid derivative (PPTN), and studied its pharma- cological properties in detail. The concentration-effect curve of UDP-glucose for promoting inhibition of adenylyl cyclase in C6 glioma cells stably expressing the P2Y14-R was shifted to the right in a concentration-dependent manner by PPTN. Schild analyses revealed that PPTN-mediated inhibition fol- lowed competitive kinetics, with a KB of 434 pM observed. In contrast, 1 mM PPTN exhibited no agonist or antagonist effect at the P2Y1, P2Y2, P2Y4, P2Y6, P2Y11, P2Y12, or P2Y13 receptors. UDP-glucose?promoted chemotaxis of differentiated HL-60 human promyelocytic leukemia cells was blocked by PPTN with a concentration dependence consistent with the KB determined with recombinant P2Y14-R. In contrast, the che- motactic response evoked by the chemoattractant peptide fMetLeuPhe was unaffected by PPTN. UDP-glucose?promoted chemotaxis of freshly isolated human neutrophils also was blocked by PPTN. In summary, this work establishes PPTN as a highly selective high-affinity antagonist of the P2Y14-R that is useful for interrogating the action of this receptor in physiologic systems.