title: High potency inhibition of hERG potassium channels by the sodium-calcium exchange inhibitor KB-R7943.
abstract: OBJECTIVE
  KB-R7943 is an isothiourea derivative that is used widely as a pharmacological inhibitor of sodium-calcium exchange (NCX) in experiments on cardiac and other tissue types. This study investigated KB-R7943 inhibition of hERG (human ether-à-go-go-related gene) K(+) channels that underpin the cardiac rapid delayed rectifier potassium current, I(Kr) .
  
  
  METHODS
  Whole-cell patch-clamp measurements were made of hERG current (I(hERG) ) carried by wild-type or mutant hERG channels and of native rabbit ventricular I(Kr) . Docking simulations utilized a hERG homology model built on a MthK-based template.
  
  
  RESULTS
  KB-R7943 inhibited both I(hERG) and native I(Kr) rapidly on membrane depolarization with IC(50) values of ∼89 and ∼120 nM, respectively, for current tails at -40 mV following depolarizing voltage commands to +20 mV. Marked I(hERG) inhibition also occurred under ventricular action potential voltage clamp. I(hERG) inhibition by KB-R7943 exhibited both time- and voltage-dependence but showed no preference for inactivated over activated channels. Results of alanine mutagenesis and docking simulations indicate that KB-R7943 can bind to a pocket formed of the side chains of aromatic residues Y652 and F656, with the compound's nitrobenzyl group orientated towards the cytoplasmic side of the channel pore. The structurally related NCX inhibitor SN-6 also inhibited I(hERG) , but with a markedly reduced potency.
  
  
  CONCLUSIONS
  KB-R7943 inhibits I(hERG) /I(Kr) with a potency that exceeds that reported previously for acute cardiac NCX inhibition. Our results also support the feasibility of benzyloxyphenyl-containing NCX inhibitors with reduced potential, in comparison with KB-R7943, to inhibit hERG.
authors.affiliation: University Walk, Bristol, UK.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Cheng
authors.forename: Yihong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Chunyun
authors.initials: C
authors.lastname: Du
authors.forename: Christopher E
authors.initials: CE
authors.lastname: Dempsey
authors.forename: Jules C
authors.initials: JC
authors.lastname: Hancox
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D001593:Benzyl Compounds / Q000494:pharmacology; D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D066298:In Vitro Techniques; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011817:Rabbits; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics; D019831:Sodium-Calcium Exchanger / Q000037:antagonists & inhibitors*; D053778:Thiazolidines / Q000494:pharmacology; D013890:Thiourea / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: C101670:2-(2-(4-(4-nitrobenzyloxy)phenyl)ethyl)isothiourea methanesulfonate; C487681:2-(4-(4-nitrobenzyloxy)benzyl)thiazolidine-4-carboxylic acid ethyl ester; D001593:Benzyl Compounds; D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; C000606913:KCNH2 protein, human; D050505:Mutant Proteins; D011994:Recombinant Proteins; D019831:Sodium-Calcium Exchanger; D053778:Thiazolidines; D013890:Thiourea