{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T19:19:24Z","timestamp":1790191164641,"version":"4.1.0"},"reference-count":58,"publisher":"Wiley","issue":"18","license":[{"start":{"date-parts":[[2015,8,4]],"date-time":"2015-08-04T00:00:00Z","timestamp":1438646400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"crossref","award":["Lo 274\/11-2"],"award-info":[{"award-number":["Lo 274\/11-2"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["bpspubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["British J Pharmacology"],"published-print":{"date-parts":[[2015,9]]},"abstract":"<jats:sec><jats:title>Background and Purpose<\/jats:title><jats:p>The<jats:styled-content style=\"fixed-case\">N<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">K<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">C<\/jats:styled-content>l cotransporters (<jats:styled-content style=\"fixed-case\">NKCCs<\/jats:styled-content>) mediate the coupled, electroneutral movement of<jats:styled-content style=\"fixed-case\"><jats:roman>Na<jats:sup>+<\/jats:sup><\/jats:roman><\/jats:styled-content>,<jats:styled-content style=\"fixed-case\"><jats:roman>K<jats:sup>+<\/jats:sup><\/jats:roman><\/jats:styled-content>and<jats:styled-content style=\"fixed-case\"><jats:roman>Cl<jats:sup>\u2212<\/jats:sup><\/jats:roman><\/jats:styled-content>ions across cell membranes. There are two isoforms of this cation co\u2010transporter,<jats:styled-content style=\"fixed-case\">NKCC<\/jats:styled-content>1 and<jats:styled-content style=\"fixed-case\">NKCC<\/jats:styled-content>2.<jats:styled-content style=\"fixed-case\">NKCC<\/jats:styled-content>2 is expressed primarily in the kidney and is the target of diuretics such as bumetanide. Bumetanide was discovered by screening \u223c5000 3\u2010amino\u20105\u2010sulfamoylbenzoic acid derivatives, long before<jats:styled-content style=\"fixed-case\">NKCC<\/jats:styled-content>2 was identified in the kidney. Therefore, structure\u2013activity studies on effects of bumetanide derivatives on<jats:styled-content style=\"fixed-case\">NKCC<\/jats:styled-content>2 are not available.<\/jats:p><\/jats:sec><jats:sec><jats:title>Experimental Approach<\/jats:title><jats:p>In this study, the effect of a series of diuretically active bumetanide derivatives was investigated on human<jats:styled-content style=\"fixed-case\">NKCC<\/jats:styled-content>2 variant A (<jats:styled-content style=\"fixed-case\">hNKCC2A<\/jats:styled-content>) expressed in<jats:styled-content style=\"fixed-case\"><jats:italic>X<\/jats:italic><\/jats:styled-content><jats:italic>enopus laevis<\/jats:italic>oocytes.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Results<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\">B<\/jats:styled-content>umetanide blocked<jats:styled-content style=\"fixed-case\">hNKCC2A<\/jats:styled-content>transport with an<jats:styled-content style=\"fixed-case\">IC<\/jats:styled-content><jats:sub>50<\/jats:sub>of 4\u2009\u03bcM. There was good correlation between the diuretic potency of bumetanide and its derivatives in dogs and their inhibition of<jats:styled-content style=\"fixed-case\">hNKCC2A<\/jats:styled-content>(<jats:italic>r<\/jats:italic><jats:sup>2<\/jats:sup>= 0.817;<jats:italic>P<\/jats:italic>&lt; 0.01). Replacement of the carboxylic group of bumetanide by a non\u2010ionic residue, for example, an anilinomethyl group, decreased inhibition of<jats:styled-content style=\"fixed-case\">hNKCC2A<\/jats:styled-content>, indicating that an acidic group was required for transporter inhibition. Exchange of the phenoxy group of bumetanide for a 4\u2010chloroanilino group or the sulfamoyl group by a methylsulfonyl group resulted in compounds with higher potency to inhibit<jats:styled-content style=\"fixed-case\">hNKCC2A<\/jats:styled-content>than bumetanide.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions and Implications<\/jats:title><jats:p>The<jats:styled-content style=\"fixed-case\"><jats:italic>X<\/jats:italic><\/jats:styled-content><jats:italic>.\u2009laevis<\/jats:italic>oocyte expression system used in these experiments allowed analysis of the structural requirements that determine relative potency of loop diuretics on human<jats:styled-content style=\"fixed-case\">NKCC2<\/jats:styled-content>splice variants, and may lead to the discovery of novel high\u2010ceiling diuretics.<\/jats:p><\/jats:sec>","DOI":"10.1111\/bph.13231","type":"journal-article","created":{"date-parts":[[2015,6,23]],"date-time":"2015-06-23T11:21:52Z","timestamp":1435058512000},"page":"4469-4480","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Structure\u2013activity relationships of bumetanide derivatives: correlation between diuretic activity in dogs and inhibition of the human<scp>NKCC<\/scp>2<scp>A<\/scp>transporter"],"prefix":"10.1111","volume":"172","author":[{"given":"Kasper","family":"Lykke","sequence":"first","affiliation":[{"name":"Department of Cellular and Molecular Medicine University of Copenhagen Copenhagen Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathrin","family":"T\u00f6llner","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Toxicology, and Pharmacy University of Veterinary Medicine Hannover Hannover Germany"},{"name":"Center for Systems Neuroscience Hannover Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kerstin","family":"R\u00f6mermann","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Toxicology, and Pharmacy University of Veterinary Medicine Hannover Hannover Germany"},{"name":"Center for Systems Neuroscience Hannover Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter W","family":"Feit","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Toxicology, and Pharmacy University of Veterinary Medicine Hannover Hannover Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Erker","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry University of Vienna Vienna Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nanna","family":"MacAulay","sequence":"additional","affiliation":[{"name":"Department of Cellular and Molecular Medicine University of Copenhagen Copenhagen Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wolfgang","family":"L\u00f6scher","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Toxicology, and Pharmacy University of Veterinary Medicine Hannover Hannover Germany"},{"name":"Center for Systems Neuroscience Hannover Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2015,8,4]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/bph.12450"},{"key":"e_1_2_10_3_1","first-page":"221","volume-title":"Cell physiology Sourcebook. Fourth Edition. Essentials of Membrane Biophysics","author":"Alvarez\u2010Leefmans FJ","year":"2012"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1528-1167.2011.03378.x"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pharmthera.2012.07.010"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1748-1716.2010.02099.x"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajprenal.00106.2008"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/j.1552-4604.1981.tb05662.x"},{"key":"e_1_2_10_9_1","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1016\/S0022-3565(25)30556-2","article-title":"A comparative diuretic and tissue distribution study of bumetanide and furosemide in the dog","volume":"197","author":"Cohen MR","year":"1976","journal-title":"J Pharmacol Exp Ther"},{"key":"e_1_2_10_10_1","doi-asserted-by":"crossref","first-page":"25677","DOI":"10.1016\/S0021-9258(18)47302-4","article-title":"Molecular cloning and chromosome localization of a putative basolateral Na(+)\u2010K(+)\u20102Cl\u2010 cotransporter from mouse inner medullary collecting duct (mIMCD\u20103) cells","volume":"269","author":"Delpire E","year":"1994","journal-title":"J Biol Chem"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/9713"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1021\/jm00287a014"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/j.1552-4604.1981.tb05661.x"},{"key":"e_1_2_10_14_1","first-page":"1","volume-title":"Bumetanide","author":"Feit PW","year":"1990"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1021\/jm00225a013"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/jm00300a012"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1021\/jm00252a002"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-009-0218-9"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-7793.1999.0699s.x"},{"issue":"6","key":"e_1_2_10_20_1","first-page":"14","article-title":"Pharmacology of bumetanide","volume":"51","author":"Frey HH","year":"1975","journal-title":"Postgrad Med J"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.1979.236.1.C1"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(17)32499-7"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pathophys.2007.09.006"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1020521308985"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00210-001-0521-y"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.3171\/FOC\/2008\/25\/9\/E22"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncpneuro0883"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrn3819"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1476-5381.2010.00872.x"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuropharm.2012.05.045"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00424-013-1370-5"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2005.06.001"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejps.2004.05.009"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1476-5381.2010.00873.x"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.1093\/med\/9780199746545.003.0045"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.1021\/bk-1978-0083.ch002"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0773.1977.tb03209.x"},{"key":"e_1_2_10_38_1","first-page":"11","article-title":"A metal complexing property of furosemide and bumetanide: determination of pK and stability constant","volume":"26","author":"Orita Y","year":"1976","journal-title":"Arzneimittelforschung"},{"key":"e_1_2_10_39_1","first-page":"66","article-title":"Pharmacological properties of 3\u2010n\u2010butylamino\u20104\u2010phenoxy\u20105\u2010sulfamylbenzoic acid (Bumetanide), a new potent diuretic","volume":"22","author":"Ostergaard EH","year":"1972","journal-title":"Arzneimittelforschung"},{"key":"e_1_2_10_40_1","doi-asserted-by":"crossref","first-page":"C139","DOI":"10.1152\/ajpcell.1980.238.3.C139","article-title":"cAMP\u2010stimulated cation cotransport in avian erythrocytes: inhibition by \u2018loop\u2019 diuretics","volume":"238","author":"Palfrey HC","year":"1980","journal-title":"Am J Physiol"},{"key":"e_1_2_10_41_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt1143"},{"key":"e_1_2_10_42_1","first-page":"G942","article-title":"Interaction of bumetanide derivatives with hepatocellular bile acid uptake","volume":"265","author":"Petzinger E","year":"1993","journal-title":"Am J Physiol"},{"issue":"1","key":"e_1_2_10_43_1","first-page":"42","article-title":"What we have learned about bumetanide and the concept of multispecific bile acid\/drug transporters from the liver","volume":"24","author":"Petzinger E","year":"1996","journal-title":"J Hepatol"},{"key":"e_1_2_10_44_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110442200"},{"key":"e_1_2_10_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.siny.2013.04.005"},{"key":"e_1_2_10_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(14)70303-5"},{"key":"e_1_2_10_47_1","doi-asserted-by":"publisher","DOI":"10.1111\/epi.12620"},{"key":"e_1_2_10_48_1","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1016\/S0030-6665(20)30770-2","article-title":"Ototoxicity of loop diuretics","volume":"26","author":"Rybak LP","year":"1993","journal-title":"Otolaryngol Clin North Am"},{"key":"e_1_2_10_49_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.ki.5002434"},{"key":"e_1_2_10_50_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)53136-7"},{"key":"e_1_2_10_51_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymeth.2009.12.001"},{"key":"e_1_2_10_52_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.356014"},{"key":"e_1_2_10_53_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.1994.266.1.C284"},{"key":"e_1_2_10_54_1","doi-asserted-by":"publisher","DOI":"10.1002\/ana.24124"},{"key":"e_1_2_10_55_1","doi-asserted-by":"publisher","DOI":"10.1111\/ejn.12424"},{"key":"e_1_2_10_56_1","doi-asserted-by":"publisher","DOI":"10.1002\/bdd.693"},{"key":"e_1_2_10_57_1","doi-asserted-by":"publisher","DOI":"10.2165\/00003495-198428050-00003"},{"key":"e_1_2_10_58_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.6.2201"},{"key":"e_1_2_10_59_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.2011.226316"}],"container-title":["British Journal of Pharmacology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fbph.13231","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/bpspubs.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/bph.13231","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,5,28]],"date-time":"2025-05-28T20:27:38Z","timestamp":1748464058000},"score":1,"resource":{"primary":{"URL":"https:\/\/bpspubs.onlinelibrary.wiley.com\/doi\/10.1111\/bph.13231"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,8,4]]},"references-count":58,"journal-issue":{"issue":"18","published-print":{"date-parts":[[2015,9]]}},"alternative-id":["10.1111\/bph.13231"],"URL":"https:\/\/doi.org\/10.1111\/bph.13231","archive":["Portico"],"relation":{},"ISSN":["0007-1188","1476-5381"],"issn-type":[{"value":"0007-1188","type":"print"},{"value":"1476-5381","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,8,4]]},"assertion":[{"value":"2015-01-15","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2015-06-12","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2015-08-04","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}