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: Matthew O Barrett, Juliana I Sesma, Christopher B Ball, P Suresh Jayasekara, Kenneth A Jacobson, Eduardo R Lazarowski, T Kendall Harden
• Journal: Molecular pharmacology (Mol Pharmacol) Vol. 84 Issue 1 Pg. 41-9 (Jul 2013) ISSN: 1521-0111 [Electronic] United States
• PMID: 23592514 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, N.I.H., Intramural)

Chemical References

• Adenylyl Cyclase Inhibitors
• P2Y14 receptor, human
• Purinergic P2 Receptor Agonists
• Purinergic P2 Receptor Antagonists
• Receptors, Purinergic P2
• Adenylyl Cyclases
• Uridine Diphosphate Glucose

Topics

• Adenylyl Cyclase Inhibitors
• Adenylyl Cyclases (metabolism)
• Animals
• CHO Cells
• Cell Line, Tumor
• Chemotaxis (drug effects)
• Cricetinae
• Glioma (metabolism)
• HL-60 Cells
• Humans
• Leukemia, Promyelocytic, Acute (metabolism)
• Neutrophils (drug effects, metabolism)
• Purinergic P2 Receptor Agonists (pharmacology)
• Purinergic P2 Receptor Antagonists (chemical synthesis, pharmacology)
• Rats
• Receptors, Purinergic P2 (metabolism)
• Uridine Diphosphate Glucose (metabolism)