Design, synthesis, pharmacological characterization of a fluorescent agonist of the P2Y14 receptor. Issue 21 (1st November 2015)

• Record Type: Journal Article
• Authors: Kiselev, Evgeny; Balasubramanian, Ramachandran; Uliassi, Elisa; Brown, Kyle A.; Trujillo, Kevin; Katritch, Vsevolod; Hammes, Eva; Stevens, Raymond C.; Harden, T. Kendall; Jacobson, Kenneth A.
• Is Part Of: Bioorganic & medicinal chemistry letters. Volume 25:Issue 21(2015)
• Page Start: 4733
• Page End: 4739
• Publication Date: 2015-11-01
• Subjects: DXQJLXWZLNCEJX-VBCGWZPHSA-N; BODIPY boron-dipyrromethene -- cAMP 3′, 5′-cyclic adenosine monophosphate -- CHO Chinese hamster ovary -- COMU [[(Z)-(1-cyano-2-ethoxy-2-oxoethylidene)amino]oxy-morpholin-4-ylmethylidene]-dimethylazanium hexafluorophosphate -- DMEM Dulbecco's modified Eagle's medium -- DMF dimethylformamide -- ECL extracellular loop -- FCM flow cytometry -- HATU 1-[bis(dimethylamino)methylene]-1H-1, 2, 3-triazolo[4, 5-b]pyridinium 3-oxid hexafluorophosphate -- IBMX 3-isobutyl-1-methylxanthine -- 2-MeSADP 2-methylthio-adenosine-5′-diphosphate -- MESF molecules of equivalent soluble fluorochrome -- MRS2578 N, N″-1, 4-butanediylbis[N'-(3-isothiocyanatophenyl)]thiourea] -- MRS2957 P1-(uridine 5′-)-P3-(N4-methoxycytidine 5′-)triphosphate -- PPTN 4-(4-(piperidin-4-yl)-phenyl)-7-(4-(trifluoromethyl)-phenyl)-2-naphthoic acid -- SAR structure activity relationship -- UDPG uridine-5′-diphosphoglucose -- TM transmembrane helix; G protein-coupled receptor -- Nucleotides -- Pyrimidines -- Homology modeling -- Docking -- P2Y receptor; Bioorganic chemistry -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1016/j.bmcl.2015.08.021
• Languages: English
• ISSNs: 0960-894X

Abstract

Graphical abstract: Abstract: The P2Y14 R is a Gi/o -coupled receptor of the P2Y family of purinergic receptors that is activated by extracellular UDP and UDP-glucose (UDPG). In an earlier report we described a P2Y14 R fluorescent probe, MRS4174, based on the potent and selective antagonist PPTN, a naphthoic acid derivative. Here, we report the design, preparation, and activity of an agonist-based fluorescent probe MRS4183 (11 ) and a shorter P2Y14 R agonist congener, which contain a UDP-glucuronic acid pharmacophore and BODIPY fluorophores conjugated through diaminoalkyl linkers. The design relied on both docking in a P2Y14 R homology model and established structure activity relationship (SAR) of nucleotide analogs.11 retained P2Y14 R potency with EC50 value of 0.96 nM (inhibition of adenylyl cyclase), compared to parent UDPG (EC50 47 nM) and served as a tracer for microscopy and flow cytometry, displaying minimal nonspecific binding. Binding saturation analysis gave an apparent binding constant for11 in whole cells of 21.4 ± 1.1 nM, with a t 1/2 of association at 50 nM11 of 23.9 min. Known P2Y14 R agonists and PPTN inhibited cell binding of11 with the expected rank order of potency. The success in the identification of a new P2Y14 R fluorescent agonist with low nonspecific binding illustrates the advantages of rational design based on recently determined GPCR X-ray structures. Such conjugates will be useful tools in expanding the SAR of this receptor, which still lacks chemical diversity in its collective ligands.