title: Inhibition and binding studies of carbonic anhydrase isozymes I, II and IX with benzimidazo[1,2-c][1,2,3]thiadiazole-7-sulphonamides.
abstract: The binding and inhibition strength of a series of benzimidazo[1,2-c][1,2,3]thiadiazole-7-sulphonamides were determined for recombinant human carbonic anhydrase isoforms I, II, and IX. The inhibition strength was determined by a stop-flow method to measure carbon dioxide hydration. Inhibitor-enzyme binding was determined by two biophysical techniques--isothermal titration calorimetry and thermal shift assay. The co-crystal structure was determined by X-ray crystallography. Comparing the results obtained using three different inhibition and binding methods increased the accuracy of compound affinity ranking and the ability to determine compound inhibitory specificity towards a particular carbonic anhydrase isoform. In most cases, all three methods yielded the same results despite using very different approaches to measure the binding and inhibition reactions. Some of the compounds studied are submicromolar inhibitors of the isoform IX, a prominent cancer target.
authors.affiliation: Laboratory of Biothermodynamics and Drug Design, Institute of Biotechnology, Graičiūno 8, Vilnius, Lithuania.
authors.forename: Lina
authors.initials: L
authors.lastname: Baranauskienė
authors.forename: Mika
authors.initials: M
authors.lastname: Hilvo
authors.forename: Jurgita
authors.initials: J
authors.lastname: Matulienė
authors.forename: Dmitrij
authors.initials: D
authors.lastname: Golovenko
authors.forename: Elena
authors.initials: E
authors.lastname: Manakova
authors.forename: Virginija
authors.initials: V
authors.lastname: Dudutienė
authors.forename: Vilma
authors.initials: V
authors.lastname: Michailovienė
authors.forename: Jolanta
authors.initials: J
authors.lastname: Torresan
authors.forename: Jelena
authors.initials: J
authors.lastname: Jachno
authors.forename: Seppo
authors.initials: S
authors.lastname: Parkkila
authors.forename: Alfonso
authors.initials: A
authors.lastname: Maresca
authors.forename: Claudiu T
authors.initials: CT
authors.lastname: Supuran
authors.forename: Saulius
authors.initials: S
authors.lastname: Gražulis
authors.forename: Daumantas
authors.initials: D
authors.lastname: Matulis
mesh_terms: D000465:Algorithms; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism; D000970:Antineoplastic Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D001562:Benzimidazoles / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D002151:Calorimetry / Q000379:methods; D024401:Carbonic Anhydrase I / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D024402:Carbonic Anhydrase II / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000071231:Carbonic Anhydrase IX; D002257:Carbonic Anhydrase Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D002256:Carbonic Anhydrases / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain / Q000187:drug effects; D018360:Crystallography, X-Ray; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008968:Molecular Conformation; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D013449:Sulfonamides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013830:Thiadiazoles / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D001562:Benzimidazoles; D002257:Carbonic Anhydrase Inhibitors; D008024:Ligands; D011994:Recombinant Proteins; D013449:Sulfonamides; D013830:Thiadiazoles; D024401:Carbonic Anhydrase I; D024402:Carbonic Anhydrase II; C089226:CA9 protein, human; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases