{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T08:11:25Z","timestamp":1767859885672,"version":"3.49.0"},"reference-count":65,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2013,12,10]],"date-time":"2013-12-10T00:00:00Z","timestamp":1386633600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["bpspubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["British J Pharmacology"],"published-print":{"date-parts":[[2014,1]]},"abstract":"<jats:sec><jats:title>Background and Purpose<\/jats:title><jats:p>Loop diuretics are widely used to inhibit the<jats:styled-content style=\"fixed-case\">N<\/jats:styled-content>a<jats:sup>+<\/jats:sup>,<jats:styled-content style=\"fixed-case\">K<\/jats:styled-content><jats:sup>+<\/jats:sup>, 2<jats:styled-content style=\"fixed-case\">C<\/jats:styled-content>l<jats:sup>\u2212<\/jats:sup>co\u2010transporter, but they also inhibit the cystic fibrosis transmembrane conductance regulator (<jats:styled-content style=\"fixed-case\">CFTR<\/jats:styled-content>)<jats:styled-content style=\"fixed-case\">C<\/jats:styled-content>l<jats:sup>\u2212<\/jats:sup>channel. Here, we investigated the mechanism of<jats:styled-content style=\"fixed-case\">CFTR<\/jats:styled-content>inhibition by loop diuretics and explored the effects of chemical structure on channel blockade.<\/jats:p><\/jats:sec><jats:sec><jats:title>Experimental Approach<\/jats:title><jats:p>Using the patch\u2010clamp technique, we tested the effects of bumetanide, furosemide, piretanide and xipamide on recombinant wild\u2010type human<jats:styled-content style=\"fixed-case\">CFTR<\/jats:styled-content>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Results<\/jats:title><jats:p>When added to the intracellular solution, loop diuretics inhibited<jats:styled-content style=\"fixed-case\">CFTR C<\/jats:styled-content>l<jats:sup>\u2212<\/jats:sup>currents with potency approaching that of glibenclamide, a widely used<jats:styled-content style=\"fixed-case\">CFTR<\/jats:styled-content>blocker with some structural similarity to loop diuretics. To begin to study the kinetics of channel blockade, we examined the time dependence of macroscopic current inhibition following a hyperpolarizing voltage step. Like glibenclamide, piretanide blockade of<jats:styled-content style=\"fixed-case\">CFTR<\/jats:styled-content>was time and voltage dependent. By contrast, furosemide blockade was voltage dependent, but time independent. Consistent with these data, furosemide blocked individual<jats:styled-content style=\"fixed-case\">CFTR C<\/jats:styled-content>l<jats:sup>\u2212<\/jats:sup>channels with \u2018very fast\u2019 speed and drug\u2010induced blocking events overlapped brief channel closures, whereas piretanide inhibited individual channels with \u2018intermediate\u2019 speed and drug\u2010induced blocking events were distinct from channel closures.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions and Implications<\/jats:title><jats:p>Structure\u2013activity analysis of the loop diuretics suggests that the phenoxy group present in bumetanide and piretanide, but absent in furosemide and xipamide, might account for the different kinetics of channel block by locking loop diuretics within the intracellular vestibule of the<jats:styled-content style=\"fixed-case\">CFTR<\/jats:styled-content>pore. We conclude that loop diuretics are open\u2010channel blockers of<jats:styled-content style=\"fixed-case\">CFTR<\/jats:styled-content>with distinct kinetics, affected by molecular dimensions and lipophilicity.<\/jats:p><\/jats:sec>","DOI":"10.1111\/bph.12458","type":"journal-article","created":{"date-parts":[[2013,10,12]],"date-time":"2013-10-12T02:20:10Z","timestamp":1381544410000},"page":"265-278","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Loop diuretics are open\u2010channel blockers of the cystic fibrosis transmembrane conductance regulator with distinct kinetics"],"prefix":"10.1111","volume":"171","author":[{"given":"Min","family":"Ju","sequence":"first","affiliation":[{"name":"School of Physiology and Pharmacology University of Bristol Bristol UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toby S","family":"Scott\u2010Ward","sequence":"additional","affiliation":[{"name":"School of Physiology and Pharmacology University of Bristol Bristol UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jia","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Physiology and Pharmacology University of Bristol Bristol UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pissared","family":"Khuituan","sequence":"additional","affiliation":[{"name":"School of Physiology and Pharmacology University of Bristol Bristol UK"},{"name":"Center of Calcium and Bone Research Department of Physiology Faculty of Science Mahidol University Bangkok Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongyu","family":"Li","sequence":"additional","affiliation":[{"name":"School of Physiology and Pharmacology University of Bristol Bristol UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhiwei","family":"Cai","sequence":"additional","affiliation":[{"name":"School of Physiology and Pharmacology University of Bristol Bristol UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen M","family":"Husbands","sequence":"additional","affiliation":[{"name":"Department of Pharmacy and Pharmacology University of Bath Bath UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David N","family":"Sheppard","sequence":"additional","affiliation":[{"name":"School of Physiology and Pharmacology University of Bristol Bristol UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2013,12,10]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/bph.12444"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s002109900096"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.1992.262.4.C803"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M108023200"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.bjp.0702748"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.200308921"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M510576200"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.00052.2013"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.2008.164970"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.072678"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00424-011-1035-1"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1021\/ci2005884"},{"key":"e_1_2_8_14_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\u2212 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_8_15_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.113332"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj20011717"},{"key":"e_1_2_8_17_1","doi-asserted-by":"crossref","first-page":"11787","DOI":"10.1016\/S0021-9258(17)44299-2","article-title":"[3H]Bumetanide binding to membranes isolated from dog kidney outer medulla: relationship to the Na,K,Cl co\u2010transport system","volume":"258","author":"Forbush B","year":"1983","journal-title":"J Biol Chem"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M708079200"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature04712"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.bjp.0704932"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.physiol.62.1.515"},{"key":"e_1_2_8_22_1","doi-asserted-by":"crossref","first-page":"C235","DOI":"10.1152\/ajpcell.1983.245.3.C235","article-title":"Bumetanide inhibits (Na + K + 2Cl) co\u2010transport at a chloride site","volume":"245","author":"Haas M","year":"1983","journal-title":"Am J Physiol Cell Physiol"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00210-001-0521-y"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00424-003-1066-3"},{"key":"e_1_2_8_25_1","volume-title":"ABC Proteins: From Bacteria to Man","author":"Holland IB","year":"2003"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.270.5239.1166"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.39.24556"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.112.5.549"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.12.7179"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.201010518"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.2165\/11313570-000000000-00000"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1523-1755.2004.00967.x"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M414354200"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1002\/jez.a.10311"},{"key":"e_1_2_8_35_1","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/S0022-3565(25)33286-6","article-title":"Nature of the inhibition of Cl\u2212 by furosemide: evidence for competitive inhibition of active transport in toad cornea","volume":"223","author":"Ludens JH","year":"1982","journal-title":"J Pharmacol Exp Ther"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI0216112"},{"key":"e_1_2_8_37_1","doi-asserted-by":"crossref","first-page":"2987","DOI":"10.1016\/S0021-9258(17)42037-0","article-title":"Stoichiometry of recombinant cystic fibrosis transmembrane conductance regulator in epithelial cells and its functional reconstitution into cells in vitro","volume":"269","author":"Marshall J","year":"1994","journal-title":"J Biol Chem"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.200409059"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi201888a"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1124\/mol.112.080267"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajprenal.00071.2005"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1185\/03007997509113658"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.1999.276.1.C231"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.2475911"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chroma.2003.09.061"},{"key":"e_1_2_8_46_1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.2000.80.1.211"},{"key":"e_1_2_8_47_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00587857"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.bjp.0707663"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajplung.1996.271.2.L192"},{"key":"e_1_2_8_50_1","doi-asserted-by":"publisher","DOI":"10.1080\/09687680310001597758"},{"key":"e_1_2_8_51_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.200409135"},{"key":"e_1_2_8_52_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-7793.1997.333bh.x"},{"key":"e_1_2_8_53_1","doi-asserted-by":"crossref","first-page":"L405","DOI":"10.1152\/ajplung.1994.266.4.L405","article-title":"Expression of cystic fibrosis transmembrane conductance regulator in a model epithelium","volume":"266","author":"Sheppard DN","year":"1994","journal-title":"Am J Physiol Lung Cell Mol Physiol"},{"key":"e_1_2_8_54_1","doi-asserted-by":"publisher","DOI":"10.1124\/mol.106.031732"},{"key":"e_1_2_8_55_1","first-page":"230","article-title":"Furosemide and bumetanide block CFTR Cl\u2212 channels","volume":"14","author":"Venglarik CJ","year":"1997","journal-title":"Pediatr Pulmonol Suppl"},{"key":"e_1_2_8_56_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(96)79839-9"},{"key":"e_1_2_8_57_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrd2780"},{"key":"e_1_2_8_58_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M503118200"},{"key":"e_1_2_8_59_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(94)80930-0"},{"key":"e_1_2_8_60_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.61.6.687"},{"key":"e_1_2_8_61_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.bjp.0704882"},{"key":"e_1_2_8_62_1","doi-asserted-by":"publisher","DOI":"10.1007\/s002320001053"},{"key":"e_1_2_8_63_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00232-004-0712-9"},{"key":"e_1_2_8_64_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00232-004-0672-0"},{"key":"e_1_2_8_65_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-7793.2001.0435f.x"},{"key":"e_1_2_8_66_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.20028685"}],"container-title":["British Journal of Pharmacology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fbph.12458","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/bpspubs.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/bph.12458","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,30]],"date-time":"2025-04-30T16:46:46Z","timestamp":1746031606000},"score":1,"resource":{"primary":{"URL":"https:\/\/bpspubs.onlinelibrary.wiley.com\/doi\/10.1111\/bph.12458"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,12,10]]},"references-count":65,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2014,1]]}},"alternative-id":["10.1111\/bph.12458"],"URL":"https:\/\/doi.org\/10.1111\/bph.12458","archive":["Portico"],"relation":{},"ISSN":["0007-1188","1476-5381"],"issn-type":[{"value":"0007-1188","type":"print"},{"value":"1476-5381","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,12,10]]},"assertion":[{"value":"2013-05-11","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2013-09-26","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2013-12-10","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}