title: Exploring a 2-naphthoic acid template for the structure-based design of P2Y14 receptor antagonist molecular probes.
abstract: The P2Y14 receptor (P2Y14R), one of eight P2Y G protein-coupled receptors (GPCR), is involved in inflammatory, endocrine, and hypoxic processes and is an attractive pharmaceutical target. The goal of this research is to develop high-affinity P2Y14R fluorescent probes based on the potent and highly selective antagonist 4-(4-(piperidin-4-yl)-phenyl)-7-(4-(trifluoromethyl)-phenyl)-2-naphthoic acid (6, PPTN). A model of hP2Y14R based on recent hP2Y12R X-ray structures together with simulated antagonist docking suggested that the piperidine ring is suitable for fluorophore conjugation while preserving affinity. Chain-elongated alkynyl or amino derivatives of 6 for click or amide coupling were synthesized, and their antagonist activities were measured in hP2Y14R-expressing CHO cells. Moreover, a new Alexa Fluor 488 (AF488) containing derivative 30 (MRS4174, Ki = 80 pM) exhibited exceptionally high affinity, as compared to 13 nM for the alkyne precursor 22. A flow cytometry assay employing 30 as a fluorescent probe was used to quantify specific binding to P2Y14R. Known P2Y receptor ligands inhibited binding of 30 with properties consistent with their previously established receptor selectivities and affinities. These results illustrate that potency in this series of 2-naphthoic acid derivatives can be preserved by chain functionalization, leading to highly potent fluorescent molecular probes for P2Y14R. Such conjugates will be useful tools in expanding the SAR of this receptor, which still lacks chemical diversity in its collective ligands. This approach demonstrates the predictive power of GPCR homology modeling and the relevance of newly determined X-ray structures to GPCR medicinal chemistry.
authors.affiliation: Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health , Bethesda, Maryland 20892, United States.
authors.forename: Evgeny
authors.initials: E
authors.lastname: Kiselev
authors.forename: Matthew O
authors.initials: MO
authors.lastname: Barrett
authors.forename: Vsevolod
authors.initials: V
authors.lastname: Katritch
authors.forename: Silvia
authors.initials: S
authors.lastname: Paoletta
authors.forename: Clarissa D
authors.initials: CD
authors.lastname: Weitzer
authors.forename: Kyle A
authors.initials: KA
authors.lastname: Brown
authors.forename: Eva
authors.initials: E
authors.lastname: Hammes
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Yin
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhao
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Stevens
authors.forename: T Kendall
authors.initials: TK
authors.lastname: Harden
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Jacobson
mesh_terms: D000818:Animals; D016466:CHO Cells; D057930:Click Chemistry; D003374:Coumarins / Q000737:chemistry; D003412:Cricetulus; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D006801:Humans; D008024:Ligands; D062105:Molecular Docking Simulation; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry*; D009281:Naphthalenes / Q000737:chemistry*; D010880:Piperidines / Q000737:chemistry; D058919:Purinergic P2 Receptor Antagonists / Q000138:chemical synthesis / Q000737:chemistry*; D018048:Receptors, Purinergic P2 / Q000737:chemistry*; D012235:Rhodamines / Q000737:chemistry; D040681:Structural Homology, Protein; D013329:Structure-Activity Relationship; D014966:Xanthenes / Q000737:chemistry
chemical_list: D003374:Coumarins; D005456:Fluorescent Dyes; D008024:Ligands; D015335:Molecular Probes; D009281:Naphthalenes; D010880:Piperidines; D058919:Purinergic P2 Receptor Antagonists; D018048:Receptors, Purinergic P2; D012235:Rhodamines; D014966:Xanthenes; C030037:2-naphthoic acid