TY - JOUR T1 - A Selective High-Affinity Antagonist of the P2Y<sub>14</sub> Receptor Inhibits UDP-Glucose–Stimulated Chemotaxis of Human Neutrophils JF - Molecular Pharmacology JO - Mol Pharmacol SP - 41 LP - 49 DO - 10.1124/mol.113.085654 VL - 84 IS - 1 AU - Matthew O. Barrett AU - Juliana I. Sesma AU - Christopher B. Ball AU - P. Suresh Jayasekara AU - Kenneth A. Jacobson AU - Eduardo R. Lazarowski AU - T. Kendall Harden Y1 - 2013/07/01 UR - http://molpharm.aspetjournals.org/content/84/1/41.abstract N2 - The nucleotide-sugar–activated P2Y14 receptor (P2Y14-R) is highly expressed in hematopoietic cells. Although the physiologic functions of this receptor remain undefined, it has been strongly implicated recently in immune and inflammatory responses. Lack of availability of receptor-selective high-affinity antagonists has impeded progress in studies of this and most of the eight nucleotide-activated P2Y receptors. A series of molecules 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 pharmacological 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 followed competitive kinetics, with a KB of 434 pM observed. In contrast, 1 μM 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 chemotactic 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. ER -