title: Synthesis, characterization, and in vitro evaluation of the selective P2Y2receptor antagonist AR-C118925.
abstract: The Gqprotein-coupled, ATP- and UTP-activated P2Y2receptor is a potential drug target for a range of different disorders, including tumor metastasis, inflammation, atherosclerosis, kidney disorders, and osteoporosis, but pharmacological studies are impeded by the limited availability of suitable antagonists. One of the most potent and selective antagonists is the thiouracil derivative AR-C118925. However, this compound was until recently not commercially available and little is known about its properties. We therefore developed an improved procedure for the synthesis of AR-C118925 and two derivatives to allow up-scaling and assessed their potency in calcium mobilization assays on the human and rat P2Y2receptors recombinantly expressed in 1321N1 astrocytoma cells. The compound was further evaluated for inhibition of P2Y2receptor-induced β-arrestin translocation. AR-C118925 behaved as a competitive antagonist with pA2values of 37.2 nM (calcium assay) and 51.3 nM (β-arrestin assay). Selectivity was assessed vs. related receptors including P2X, P2Y, and adenosine receptor subtypes, as well as ectonucleotidases. AR-C118925 showed at least 50-fold selectivity against the other investigated targets, except for the P2X1 and P2X3 receptors which were blocked by AR-C118925 at concentrations of about 1 μM. AR-C118925 is soluble in buffer at pH 7.4 (124 μM) and was found to be metabolically highly stable in human and mouse liver microsomes. In Caco2 cell experiments, the compound displayed moderate permeability indicating that it may show limited peroral bioavailability. AR-C118925 appears to be a useful pharmacological tool for in vitro and in vivo studies.
authors.affiliation: PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical Sciences Bonn (PSB), Pharmaceutical Chemistry I, University of Bonn, Bonn, Germany.|Pharmazeutisches Institut, Pharmazeutische Chemie I, An der Immenburg 4, D-53121, Bonn, Germany.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Rafehi
authors.affiliation: PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical Sciences Bonn (PSB), Pharmaceutical Chemistry I, University of Bonn, Bonn, Germany.
authors.forename: Joachim C
authors.initials: JC
authors.lastname: Burbiel
authors.affiliation: PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical Sciences Bonn (PSB), Pharmaceutical Chemistry I, University of Bonn, Bonn, Germany.|Pharmazeutisches Institut, Pharmazeutische Chemie I, An der Immenburg 4, D-53121, Bonn, Germany.
authors.forename: Isaac Y
authors.initials: IY
authors.lastname: Attah
authors.affiliation: PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical Sciences Bonn (PSB), Pharmaceutical Chemistry I, University of Bonn, Bonn, Germany.|Pharmazeutisches Institut, Pharmazeutische Chemie I, An der Immenburg 4, D-53121, Bonn, Germany.
authors.forename: Aliaa
authors.initials: A
authors.lastname: Abdelrahman
authors.affiliation: PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical Sciences Bonn (PSB), Pharmaceutical Chemistry I, University of Bonn, Bonn, Germany. christa.mueller@uni-bonn.de.|Pharmazeutisches Institut, Pharmazeutische Chemie I, An der Immenburg 4, D-53121, Bonn, Germany. christa.mueller@uni-bonn.de.
authors.forename: Christa E
authors.initials: CE
authors.lastname: Müller
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D003986:Dibenzocycloheptenes / Q000138:chemical synthesis* / Q000494:pharmacology*; D006801:Humans; D021381:Protein Transport; D058921:Purinergic P2Y Receptor Antagonists / Q000138:chemical synthesis* / Q000494:pharmacology*; D011744:Pyrimidinones / Q000138:chemical synthesis* / Q000494:pharmacology*; D051381:Rats; D058473:Receptors, Purinergic P2Y2 / Q000378:metabolism; D000071557:beta-Arrestins / Q000378:metabolism
chemical_list: C000624532:AR-C118925; D003986:Dibenzocycloheptenes; D058921:Purinergic P2Y Receptor Antagonists; D011744:Pyrimidinones; D058473:Receptors, Purinergic P2Y2; D000071557:beta-Arrestins
keywords: 1321N1 astrocytoma cells; AR-C118925; Antagonist; G protein-coupled receptor; P2 receptor; P2Y2 receptor; Synthesis; Thiouracil derivative