title: A selective cysteinyl leukotriene receptor 2 antagonist blocks myocardial ischemia/reperfusion injury and vascular permeability in mice.
abstract: Cysteinyl leukotrienes (CysLTs) are potent inflammatory mediators that predominantly exert their effects by binding to cysteinyl leukotriene receptors of the G protein-coupled receptor family. CysLT receptor 2 (CysLT(2)R), expressed in endothelial cells of some vascular beds, has been implicated in a variety of cardiovascular functions. Endothelium-specific overexpression of human CysLT(2)R in transgenic mice (hEC-CysLT(2)R) greatly increases myocardial infarction damage. Investigation of this receptor, however, has been hindered by the lack of selective pharmacological antagonists. Here, we describe the characterization of 3-(((3-carboxycyclohexyl)amino)carbonyl)-4-(3-(4-(4-phenoxybutoxy)phenyl)-propoxy)benzoic acid (BayCysLT(2)) and explore the selective effects of this compound in attenuating myocardial ischemia/reperfusion damage and vascular leakage. Using a recently developed β-galactosidase-β-arrestin complementation assay for CysLT(2)R activity (Mol Pharmacol 79:270-278, 2011), we determined BayCysLT(2) to be ∼20-fold more potent than the nonselective dual CysLT receptor 1 (CysLT(1)R)/CysLT(2)R antagonist 4-(((1R,2E,4E,6Z,9Z)-1-((1S)-4-carboxy-1-hydroxybutyl)-2,4,6,9-pentadecatetraen-1-yl)thio)benzoic acid (Bay-u9773) (IC(50) 274 nM versus 4.6 μM, respectively). Intracellular calcium mobilization in response to cysteinyl leukotriene administration showed that BayCysLT(2) was >500-fold more selective for CysLT(2)R compared with CysLT(1)R. Intraperitoneal injection of BayCysLT(2) in mice significantly attenuated leukotriene D(4)-induced Evans blue dye leakage in the murine ear vasculature. BayCysLT(2) administration either before or after ischemia/reperfusion attenuated the aforementioned increased myocardial infarction damage in hEC-CysLT(2)R mice. Finally, decreased neutrophil infiltration and leukocyte adhesion molecule mRNA expression were observed in mice treated with antagonist compared with untreated controls. In conclusion, we present the characterization of a potent and selective antagonist for CysLT(2)R that is useful for discerning biological activities of this receptor.
authors: Department of Biomedical and Molecular Sciences, Queen’s University, Kingston, Ontario, Canada.
authors: Nathan C
authors: NC
authors: Ni
authors: Dong
authors: D
authors: Yan
authors: Laurel L
authors: LL
authors: Ballantyne
authors: Alma
authors: A
authors: Barajas-Espinosa
authors: Tim
authors: T
authors: St Amand
authors: Derek A
authors: DA
authors: Pratt
authors: Colin D
authors: CD
authors: Funk
mesh_terms: D000818:Animals; D019390:Arrestins / Q000032:analysis; D002199:Capillary Permeability / Q000187:drug effects*; D003509:Cyclohexanecarboxylic Acids / Q000494:pharmacology*; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical; D004423:Ear / Q000098:blood supply; D006801:Humans; D020024:Leukotriene Antagonists / Q000494:pharmacology*; D017998:Leukotriene D4 / Q000037:antagonists & inhibitors*; D051379:Mice; D008822:Mice, Transgenic; D009203:Myocardial Infarction / Q000188:drug therapy* / Q000378:metabolism; D015428:Myocardial Reperfusion Injury / Q000188:drug therapy* / Q000503:physiopathology; D009206:Myocardium / Q000473:pathology; D032383:Myocytes, Cardiac / Q000187:drug effects; D009195:Peroxidase / Q000378:metabolism; D010795:Phthalic Acids / Q000494:pharmacology*; D018077:Receptors, Leukotriene / Q000378:metabolism*; D013189:SRS-A / Q000031:analogs & derivatives* / Q000494:pharmacology; D000071557:beta-Arrestins; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: C569223:3-(((3-carboxycyclohexyl)amino)carbonyl)-4-(3-(4-(4-phenoxybutoxy)phenyl)propoxy)benzoic acid; D019390:Arrestins; C071435:BAY u9773; D003509:Cyclohexanecarboxylic Acids; D020024:Leukotriene Antagonists; D010795:Phthalic Acids; D018077:Receptors, Leukotriene; D013189:SRS-A; D000071557:beta-Arrestins; D017998:Leukotriene D4; C412375:cysteinyl leukotriene receptor 2; D009195:Peroxidase; D001616:beta-Galactosidase