title: A selective high-affinity antagonist of the P2Y14 receptor inhibits UDP-glucose-stimulated chemotaxis of human neutrophils.
abstract: 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.
authors.affiliation: Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, NC 27599, USA.
authors.forename: Matthew O
authors.initials: MO
authors.lastname: Barrett
authors.forename: Juliana I
authors.initials: JI
authors.lastname: Sesma
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Ball
authors.forename: P Suresh
authors.initials: PS
authors.lastname: Jayasekara
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Jacobson
authors.forename: Eduardo R
authors.initials: ER
authors.lastname: Lazarowski
authors.forename: T Kendall
authors.initials: TK
authors.lastname: Harden
mesh_terms: D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D016466:CHO Cells; D045744:Cell Line, Tumor; D002633:Chemotaxis / Q000187:drug effects*; D006224:Cricetinae; D005910:Glioma / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D058910:Purinergic P2 Receptor Agonists / Q000494:pharmacology; D058919:Purinergic P2 Receptor Antagonists / Q000138:chemical synthesis / Q000494:pharmacology*; D051381:Rats; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D014532:Uridine Diphosphate Glucose / Q000378:metabolism*
chemical_list: D000067956:Adenylyl Cyclase Inhibitors; C518528:P2Y14 receptor, human; D058910:Purinergic P2 Receptor Agonists; D058919:Purinergic P2 Receptor Antagonists; D018048:Receptors, Purinergic P2; D000262:Adenylyl Cyclases; D014532:Uridine Diphosphate Glucose