65623 20210121114546.0 doi 10.1124/mol.114.095745 sideral 89292 ART-2015-89292 eng (orcid)0000-0001-5348-924X Oliván-Viguera, Aida A novel pan-negative-gating modulator of KCa2/3 channels, fluoro-di-benzoate, RA-2, inhibits Endothelium-derived hyperpolarization–type relaxation in coronary artery and produces bradycardia in vivo 2015 Access copy available to the general public Unrestricted Small/intermediate conductance KCa channels (KCa2/3) are Ca2+/calmodulin regulated K+ channels that produce membrane hyperpolarization and shape neurologic, epithelial, cardiovascular, and immunologic functions. Moreover, they emerged as therapeutic targets to treat cardiovascular disease, chronic inflammation, and some cancers. Here, we aimed to generate a new pharmacophore for negative-gating modulation of KCa2/3 channels. We synthesized a series of mono- and dibenzoates and identified three dibenzoates [1,3-phenylenebis(methylene) bis(3-fluoro-4-hydroxybenzoate) (RA-2), 1,2-phenylenebis(methylene) bis(3-fluoro-4-hydroxybenzoate), and 1,4-phenylenebis(methylene) bis(3-fluoro-4-hydroxybenzoate)] with inhibitory efficacy as determined by patch clamp. Among them, RA-2 was the most drug-like and inhibited human KCa3.1 with an IC50 of 17 nM and all three human KCa2 subtypes with similar potencies. RA-2 at 100 nM right-shifted the KCa3.1 concentration-response curve for Ca2+ activation. The positive-gating modulator naphtho[1,2-d]thiazol-2-ylamine (SKA-31) reversed channel inhibition at nanomolar RA-2 concentrations. RA-2 had no considerable blocking effects on distantly related large-conductance KCa1.1, Kv1.2/1.3, Kv7.4, hERG, or inwardly rectifying K+ channels. In isometric myography on porcine coronary arteries, RA-2 inhibited bradykinin-induced endothelium-derived hyperpolarization (EDH)–type relaxation in U46619-precontracted rings. Blood pressure telemetry in mice showed that intraperitoneal application of RA-2 (=100 mg/kg) did not increase blood pressure or cause gross behavioral deficits. However, RA-2 decreased heart rate by ˜145 beats per minute, which was not seen in KCa3.1-/- mice. In conclusion, we identified the KCa2/3–negative-gating modulator, RA-2, as a new pharmacophore with nanomolar potency. RA-2 may be of use to generate structurally new types of negative-gating modulators that could help to define the physiologic and pathomechanistic roles of KCa2/3 in the vasculature, central nervous system, and during inflammation in vivo. info:eu-repo/grantAgreement/ES/ISCIII/RD12-0042-0014 info:eu-repo/grantAgreement/EC/FP7/321721/EU/Identification and validation of cerebral KCa3.1/KCa2.3 potassium channels a drug tragets for the prevention and treatment of cerebral ischemia associated with diabetes and Alzheimers disease/BRAINIK info:eu-repo/grantAgreement/ES/DGA/GIPASC-B105 info:eu-repo/grantAgreement/ES/DGA/GAE-102 info:eu-repo/semantics/openAccess All rights reserved http://www.europeana.eu/rights/rr-f/ 3.931 2015 PHARMACOLOGY & PHARMACY 45 / 255 = 0.176 2015 Q1 T1 2.037 2015 Pharmacology 2015 Q1 Molecular Medicine 2015 Q1 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion (orcid)0000-0002-2231-7565 Valero, Marta Sofía Coleman, Nicole Brown, Brandon M. Laría, Celia (orcid)0000-0002-3917-4740 Murillo, María Divina Universidad de Zaragoza (orcid)0000-0002-4972-7476 Gálvez, José A. Universidad de Zaragoza (orcid)0000-0001-9033-8459 Díaz-de-Villegas, María D. Wulff, Heike (orcid)0000-0002-5554-3040 Badorrey, Ramón Universidad de Zaragoza Köhler, Ralf 1005 410 Universidad de Zaragoza Dpto. Farmacología y Fisiolog. Área Fisiología 2013 765 Universidad de Zaragoza Dpto. Química Orgánica Área Química Orgánica 87, 2 (2015), 338-348 Mol. pharmacol. Molecular Pharmacology 0026-895X 807358 http://zaguan.unizar.es/record/65623/files/texto_completo.pdf Versión publicada 141563 http://zaguan.unizar.es/record/65623/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:65623 articulos driver 2021-01-21-11:19:37 ARTICLE