{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T18:11:02Z","timestamp":1790878262260,"version":"4.1.0"},"reference-count":26,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,1,16]],"date-time":"2023-01-16T00:00:00Z","timestamp":1673827200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Recovery and Resilience Plan (NRRP)"},{"name":"European Union\u2014NextGenerationEU"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>This paper reports an investigation into the impact of pyridyl functional groups in conjunction with hydroxide-substituted benzenesulfonamides on the inhibition of human carbonic anhydrase (CA; EC 4.2.1.1) enzymes. These compounds were tested in vitro of CA II and CA IX, two physiologically important CA isoforms. The most potent inhibitory molecules against CA IX, 3g, 3h, and 3k, were studied to understand their binding modes via X-ray crystallography in adduct with CA II and CA IX-mimic. This research further adds to the field of CA inhibitors to better understand ligand selectivity between isoforms found in humans.<\/jats:p>","DOI":"10.3390\/molecules28020890","type":"journal-article","created":{"date-parts":[[2023,1,16]],"date-time":"2023-01-16T05:30:07Z","timestamp":1673847007000},"page":"890","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["New Insights into Conformationally Restricted Carbonic Anhydrase Inhibitors"],"prefix":"10.3390","volume":"28","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5657-4773","authenticated-orcid":false,"given":"Jacob","family":"Combs","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, FL 32610, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5010-0355","authenticated-orcid":false,"given":"Murat","family":"Bozdag","sequence":"additional","affiliation":[{"name":"NEUROFARBA Department, Sezione di Scienze Farmaceutiche e Nutraceutiche, University of Florence, Via Ugo Schiff 6, 50019 Florence, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lochlin D.","family":"Cravey","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, FL 32610, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anusha","family":"Kota","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, FL 32610, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2792-935X","authenticated-orcid":false,"given":"Robert","family":"McKenna","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, FL 32610, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1470-7192","authenticated-orcid":false,"given":"Andrea","family":"Angeli","sequence":"additional","affiliation":[{"name":"NEUROFARBA Department, Sezione di Scienze Farmaceutiche e Nutraceutiche, University of Florence, Via Ugo Schiff 6, 50019 Florence, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1141-6146","authenticated-orcid":false,"given":"Fabrizio","family":"Carta","sequence":"additional","affiliation":[{"name":"NEUROFARBA Department, Sezione di Scienze Farmaceutiche e Nutraceutiche, University of Florence, Via Ugo Schiff 6, 50019 Florence, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4262-0323","authenticated-orcid":false,"given":"Claudiu T.","family":"Supuran","sequence":"additional","affiliation":[{"name":"NEUROFARBA Department, Sezione di Scienze Farmaceutiche e Nutraceutiche, University of Florence, Via Ugo Schiff 6, 50019 Florence, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"6328","DOI":"10.1021\/acs.jmedchem.8b00770","article-title":"Discovery of 4-Hydroxy-3-(3-(Phenylureido)Benzenesulfonamides as SLC-0111 Analogues for the Treatment of Hypoxic Tumors Overexpressing Carbonic Anhydrase IX","volume":"61","author":"Bozdag","year":"2018","journal-title":"J. Med. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1002\/ijc.10212","article-title":"Efficient Tumor Targeting by Single-Domain Antibody Fragments of Camels","volume":"98","author":"Lauwereys","year":"2002","journal-title":"Int. J. Cancer"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1080\/14756366.2021.2004593","article-title":"First Studies on Tumor Associated Carbonic Anhydrases IX and XII Monoclonal Antibodies Conjugated to Small Molecule Inhibitors","volume":"37","author":"Testa","year":"2022","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1111\/j.1432-1033.1975.tb02449.x","article-title":"The Catalytic Mechanism of Carbonic Anhydrase","volume":"59","author":"Steiner","year":"1975","journal-title":"Eur. J. Biochem."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Frost, S.C., and McKenna, R. (2013). Carbonic Anhydrase: Mechanism, Regulation, Links to Disease, and Industrial Applications. Subcellular Biochemistry, Springer.","DOI":"10.1007\/978-94-007-7359-2"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4421","DOI":"10.1021\/cr200176r","article-title":"Multiple Binding Modes of Inhibitors to Carbonic Anhydrases: How to Design Specific Drugs Targeting 15 Different Isoforms?","volume":"112","author":"Alterio","year":"2012","journal-title":"Chem. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"8371","DOI":"10.1039\/c0cc02707c","article-title":"Selective Hydrophobic Pocket Binding Observed within the Carbonic Anhydrase II Active Site Accommodate Different 4-Substituted-Ureido-Benzenesulfonamides and Correlate to Inhibitor Potency","volume":"46","author":"Pacchiano","year":"2010","journal-title":"Chem. Commun."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1713","DOI":"10.1021\/acs.jmedchem.0c02077","article-title":"Inhibition of Carbonic Anhydrase Using SLC-149: Support for a Noncatalytic Function of CAIX in Breast Cancer","volume":"64","author":"Mboge","year":"2021","journal-title":"J. Med. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"7422","DOI":"10.1021\/acs.jmedchem.0c00733","article-title":"Sulfonamide Inhibitors of Human Carbonic Anhydrases Designed through a Three-Tails Approach: Improving Ligand\/Isoform Matching and Selectivity of Action","volume":"63","author":"Bonardi","year":"2020","journal-title":"J. Med. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1007\/s10555-019-09799-0","article-title":"The Role of Carbonic Anhydrase IX in Cancer Development: Links to Hypoxia, Acidosis, and Beyond","volume":"38","author":"Pastorekova","year":"2019","journal-title":"Cancer Metastasis Rev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"659332","DOI":"10.3389\/fonc.2021.659332","article-title":"Tissue Expression of Carbonic Anhydrase IX Correlates to More Aggressive Phenotype of Basal Cell Carcinoma","volume":"11","author":"Russo","year":"2021","journal-title":"Front. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e202200148","DOI":"10.1002\/cmdc.202200148","article-title":"Small Molecule Alkoxy Oriented Selectiveness on Human Carbonic Anhydrase II and IX Inhibition","volume":"17","author":"Bozdag","year":"2022","journal-title":"Chem. Med. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"7392","DOI":"10.1021\/acs.jmedchem.0c00636","article-title":"Azidothymidine \u201cClicked\u201d into 1,2,3-Triazoles: First Report on Carbonic Anhydrase\u2013Telomerase Dual-Hybrid Inhibitors","volume":"63","author":"Berrino","year":"2020","journal-title":"J. Med. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2561","DOI":"10.1016\/S0021-9258(18)62326-9","article-title":"The Carbon Dioxide Hydration Activity of Carbonic Anhydrase. I. Stop-Flow Kinetic Studies on the Native Human Isoenzymes B and C","volume":"246","author":"Khalifah","year":"1971","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1322","DOI":"10.1021\/bi802035f","article-title":"Design of a Carbonic Anhydrase IX Active-Site Mimic To Screen Inhibitors for Possible Anticancer Properties","volume":"48","author":"Genis","year":"2009","journal-title":"Biochemistry"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"7210","DOI":"10.1016\/j.bmc.2013.08.033","article-title":"Structural Study of Interaction between Brinzolamide and Dorzolamide Inhibition of Human Carbonic Anhydrases","volume":"21","author":"Pinard","year":"2013","journal-title":"Bioorg. Med. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e453543","DOI":"10.1155\/2015\/453543","article-title":"Probing the Surface of Human Carbonic Anhydrase for Clues towards the Design of Isoform Specific Inhibitors","volume":"2015","author":"Pinard","year":"2015","journal-title":"BioMed Res. Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1042\/bj2170727","article-title":"Carbonic Anhydrase in Human Platelets","volume":"217","author":"Siffert","year":"1984","journal-title":"Biochem. J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1021\/ar00145a005","article-title":"The Catalytic Mechanism of Carbonic Anhydrase: Implications of a Rate-Limiting Protolysis of Water","volume":"21","author":"Silverman","year":"1988","journal-title":"Acc. Chem. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1107\/S0907444909047337","article-title":"XDS","volume":"66","author":"Kabsch","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1107\/S0907444910045749","article-title":"Overview of the CCP4 Suite and Current Developments","volume":"67","author":"Winn","year":"2011","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2930","DOI":"10.1021\/bi062066y","article-title":"Atomic Crystal and Molecular Dynamics Simulation Structures of Human Carbonic Anhydrase II:\u2009 Insights into the Proton Transfer Mechanism","volume":"46","author":"Fisher","year":"2007","journal-title":"Biochemistry"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1107\/S0907444909052925","article-title":"PHENIX: A Comprehensive Python-Based System for Macromolecular Structure Solution","volume":"66","author":"Adams","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2126","DOI":"10.1107\/S0907444904019158","article-title":"Coot: Model-Building Tools for Molecular Graphics","volume":"60","author":"Emsley","year":"2004","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_25","unstructured":"(2023, January 09). The PyMOL Molecular Graphics System, Available online: pymol.org."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2778","DOI":"10.1021\/ci200227u","article-title":"LigPlot+: Multiple Ligand\u2013Protein Interaction Diagrams for Drug Discovery","volume":"51","author":"Laskowski","year":"2011","journal-title":"J. Chem. Inf. Model."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/28\/2\/890\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:07:12Z","timestamp":1760119632000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/28\/2\/890"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,16]]},"references-count":26,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["molecules28020890"],"URL":"https:\/\/doi.org\/10.3390\/molecules28020890","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,16]]}}}