{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T13:01:39Z","timestamp":1777640499934,"version":"3.51.4"},"reference-count":43,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2026,1,14]],"date-time":"2026-01-14T00:00:00Z","timestamp":1768348800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Recovery and Resilience Facility","award":["40\/OSI\/ZG"],"award-info":[{"award-number":["40\/OSI\/ZG"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background: Among the 15 human (h) carbonic anhydrase (CA; EC 4.2.1.1) isoforms, hCA IX and XII are particularly important due to their roles in tumor cell growth and survival, identifying them as promising targets for anticancer therapy. As a result, considerable effort has been directed toward the development of novel inhibitors that are highly selective for these isoforms. Methods: A library of twelve novel N-aryl-2-(4-sulfamoylphenyl)hydrazine-1-carbothioamides 3 along with two new N-aryl-2-(4-sulfamoylphenyl)hydrazine-1-carboxamide derivatives 5 were synthesized and their inhibition abilities were tested against four human carbonic anhydrase isozymes (hCA I, II, IX and XII) related to some global diseases including glaucoma, cancer and osteoporosis. Results: All compounds exhibited potent inhibition of the tested isoforms in the nanomolar range. Compound 3i showed the highest inhibition of hCA I activity but demonstrated poor selectivity toward the other isoforms. Compound 3h displayed superior selectivity for hCA II over hCA I (hCA I\/II = 37) and exhibited 2.5-fold higher inhibitory activity compared to acetazolamide (AAZ). Among the tested compounds, 3l (Ki = 32.1 nM) demonstrated markedly improved selectivity for hCA IX over hCA I, II, and XII relative to the standard drug. Notably, compound 3a showed the most potent inhibition against hCA XII (Ki = 6.8 nM), comparable to AAZ, while exhibiting significantly greater selectivity over off-target isoforms and the other tumor-associated isozyme (hCA IX\/XII = 20 versus hCA IX\/XII = 4.5 for AAZ). Conclusions: The present study suggests potent lead compounds as selective hCA IX and XII inhibitors with anticancer activity.<\/jats:p>","DOI":"10.3390\/ph19010151","type":"journal-article","created":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T08:26:36Z","timestamp":1768465596000},"page":"151","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Investigation on N-Aryl-2-(4-sulfamoylphenyl)hydrazine-1-carbothioamide as Human Carbonic Anhydrases Inhibitors"],"prefix":"10.3390","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1647-4262","authenticated-orcid":false,"given":"Morteza","family":"Abdoli","sequence":"first","affiliation":[{"name":"Latvian Institute of Organic Synthesis, LV-1006 Riga, Latvia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1470-7192","authenticated-orcid":false,"given":"Andrea","family":"Angeli","sequence":"additional","affiliation":[{"name":"Department of Neurofarba, Universit\u00e0 degli Studi di Firenze, 50019 Florence, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5520-740X","authenticated-orcid":false,"given":"Alessandro","family":"Bonardi","sequence":"additional","affiliation":[{"name":"Department of Neurofarba, Universit\u00e0 degli Studi di Firenze, 50019 Florence, Italy"},{"name":"Pharmaceutical and Nutraceutical Section and Laboratory of Molecular Modeling Cheminformatics & QSAR, Department of NEUROFARBA, University of Florence, Via U. Schiff 6, Sesto Fiorentino, 50019 Florence, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9137-2509","authenticated-orcid":false,"given":"Paola","family":"Gratteri","sequence":"additional","affiliation":[{"name":"Pharmaceutical and Nutraceutical Section and Laboratory of Molecular Modeling Cheminformatics & QSAR, Department of NEUROFARBA, University of Florence, Via U. Schiff 6, Sesto Fiorentino, 50019 Florence, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ludmila","family":"Jackevica","sequence":"additional","affiliation":[{"name":"Latvian Institute of Organic Synthesis, LV-1006 Riga, Latvia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6495-6317","authenticated-orcid":false,"given":"Antons","family":"Sizovs","sequence":"additional","affiliation":[{"name":"Latvian Institute of Organic Synthesis, LV-1006 Riga, Latvia"},{"name":"Baltic Biomaterials Centre of Excellence, Headquarters at Riga Technical University, LV-1048 Riga, Latvia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4262-0323","authenticated-orcid":false,"given":"Claudiu T.","family":"Supuran","sequence":"additional","affiliation":[{"name":"Department of Neurofarba, Universit\u00e0 degli Studi di Firenze, 50019 Florence, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9471-1342","authenticated-orcid":false,"given":"Raivis","family":"\u017dalubovskis","sequence":"additional","affiliation":[{"name":"Latvian Institute of Organic Synthesis, LV-1006 Riga, Latvia"},{"name":"Institute of Chemistry and Chemical Technology, Faculty of Natural Sciences and Technology, Riga Technical University, LV-1048 Riga, Latvia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.plantsci.2017.12.002","article-title":"The many types of carbonic anhydrases in photosynthetic organisms","volume":"268","author":"DiMario","year":"2018","journal-title":"Plant Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"453543","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_3","doi-asserted-by":"crossref","first-page":"1570","DOI":"10.1016\/j.bmc.2012.04.044","article-title":"Structure, function and applications of carbonic anhydrase isozymes","volume":"21","author":"Hassan","year":"2013","journal-title":"Bioorg. Med. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1327","DOI":"10.1152\/physrev.00018.2021","article-title":"Carbonic anhydrases in metazoan model organisms: Molecules, mechanisms, and physiology","volume":"102","author":"Aspatwar","year":"2022","journal-title":"Physiol. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1080\/13543776.2019.1629419","article-title":"Carbonic anhydrases as disease markers","volume":"29","author":"Zamanova","year":"2019","journal-title":"Expert Opin. Ther. Pat."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1038\/nrd2467","article-title":"Carbonic anhydrases: Novel therapeutic applications for inhibitors and activators","volume":"7","author":"Supuran","year":"2008","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"McDonald, P.C., Chafe, S.C., Supuran, C.T., and Dedhar, S. (2022). Cancer Therapeutic Targeting of Hypoxia Induced Carbonic Anhydrase IX: From Bench to Bedside. Cancers, 14.","DOI":"10.3390\/cancers14143297"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1080\/13543776.2022.2083502","article-title":"Anticancer carbonic anhydrase inhibitors: A patent and literature update 2018\u20132022","volume":"32","author":"Nerella","year":"2022","journal-title":"Expert Opin. Ther. Pat."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1149","DOI":"10.4155\/fmc.14.68","article-title":"Sulfonamides and their isosters as carbonic anhydrase inhibitors","volume":"6","author":"Carta","year":"2014","journal-title":"Future Med. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1007\/s11906-014-0467-3","article-title":"New insights into carbonic anhydrase inhibition, vasodilation, and treatment of hypertensive-related diseases","volume":"16","author":"Swenson","year":"2014","journal-title":"Curr. Hypertens. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e202200658","DOI":"10.1002\/cmdc.202200658","article-title":"Inhibition Studies on Carbonic Anhydrase Isoforms I, II, IX, and XII with a Series of Sulfaguanidines","volume":"18","author":"Abdoli","year":"2023","journal-title":"ChemMedChem"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"976","DOI":"10.1016\/j.bmc.2016.01.019","article-title":"Novel 3,4-diarylpyrazolines as potent cannabinoid CB1 receptor antagonists with lower lipophilicity","volume":"24","author":"Lomelino","year":"2016","journal-title":"Bioorg. Med. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"103123","DOI":"10.1016\/j.bioorg.2019.103123","article-title":"Ureidobenzenesulfonamides as efficient inhibitors of carbonic anhydrase II","volume":"91","author":"Serbian","year":"2019","journal-title":"Bioorg. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Abdoli, M., Bonardi, A., Paoletti, N., Aspatwar, A., Parkkila, S., Gratteri, P., Supuran, C.T., and \u017dalubovskis, R. (2023). Inhibition Studies on Human and Mycobacterial Carbonic Anhydrases with N-((4-Sulfamoylphenyl)carbamothioyl) Amides. Molecules, 28.","DOI":"10.3390\/molecules28104020"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2152811","DOI":"10.1080\/14756366.2022.2152811","article-title":"Investigation of novel alkyl\/benzyl (4-sulphamoylphenyl)carbamimidothioates as carbonic anhydrase inhibitors","volume":"38","author":"Abdoli","year":"2023","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5466","DOI":"10.1016\/j.bmcl.2008.09.030","article-title":"Carbonic anhydrase inhibitors: Inhibition of Plasmodium falciparum carbonic anhydrase with aromatic\/heterocyclic sulfonamides\u2014In vitro and in vivo studies","volume":"18","author":"Krungkrai","year":"2008","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"4834","DOI":"10.1021\/jm0501073","article-title":"Carbonic anhydrase inhibitors. Design of fluorescent sulfonamides as probes of tumor-associated carbonic anhydrase IX that inhibit isozyme IX-mediated acidification of hypoxic tumors","volume":"48","author":"Cecchi","year":"2005","journal-title":"J. Med. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1080\/14756366.2018.1532419","article-title":"The carbonic anhydrase IX inhibitor SLC-0111 sensitises cancer cells to conventional chemotherapy","volume":"34","author":"Andreucci","year":"2019","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1097\/COC.0000000000000691","article-title":"A Phase 1 Study of SLC-0111, a Novel Inhibitor of Carbonic Anhydrase IX, in Patients With Advanced Solid Tumors","volume":"43","author":"McDonald","year":"2020","journal-title":"Am. J. Clin. Oncol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Eloranta, K., Pihlajoki, M., Liljestr\u00f6m, E., Nousiainen, R., Soini, T., Lohi, J., Cairo, S., Wilson, D.B., Parkkila, S., and Heikinheimo, M. (2023). SLC-0111, an inhibitor of carbonic anhydrase IX, attenuates hepatoblastoma cell viability and migration. Front. Oncol., 13.","DOI":"10.3389\/fonc.2023.1118268"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Abdoli, M., De Luca, V., Capasso, C., Supuran, C.T., and \u017dalubovskis, R. (2022). Benzenesulfonamides Incorporating Hydantoin Moieties Effectively Inhibit Eukaryoticand Human Carbonic Anhydrases. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232214115"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1568","DOI":"10.1080\/14756366.2022.2080816","article-title":"4-(3-Alkyl\/benzyl-guanidino)benzenesulfonamides as selective carbonic anhydrase VII inhibitors","volume":"37","author":"Abdoli","year":"2022","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1080\/14756366.2022.2138367","article-title":"4-Cyanamidobenzenesulfonamide derivatives: A novel class of human and bacterial carbonic anhydrase inhibitors","volume":"38","author":"Abdoli","year":"2023","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2163243","DOI":"10.1080\/14756366.2022.2163243","article-title":"Novel thiazolone-benzenesulphonamide inhibitors of human and bacterial carbonic anhydrases","volume":"38","author":"Abdoli","year":"2023","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"106725","DOI":"10.1016\/j.bioorg.2023.106725","article-title":"4-Cyanamido-substituted benzenesulfonamides act as dual carbonic anhydrase and cathepsin inhibitors","volume":"139","author":"Abdoli","year":"2023","journal-title":"Bioorg. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1067","DOI":"10.1021\/acsmedchemlett.3c00200","article-title":"Inhibition Profiles of Some Novel Sulfonamide-Incorporated \u03b1-Aminophosphonates on Human Carbonic Anhydrases","volume":"14","author":"Sobati","year":"2023","journal-title":"ACS Med. Chem. Lett."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"615","DOI":"10.4155\/fmc-2022-0287","article-title":"Investigation of carbonic anhydrase inhibitory potency of (Z\/E)-alkyl N \u2032-benzyl-N-(arylsulfonyl)-carbamimidothioates","volume":"15","author":"Abdoli","year":"2023","journal-title":"Future Med. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e2400038","DOI":"10.1002\/ardp.202400038","article-title":"Sulfonamide-incorporated bis(\u03b1-aminophosphonates) as promising carbonic anhydrase inhibitors: Design, synthesis, biological evaluation, and X-ray crystallographic studies","volume":"357","author":"Sobati","year":"2024","journal-title":"Arch. Pharm."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e202400197","DOI":"10.1002\/cmdc.202400197","article-title":"Synthesis and Carbonic Anhydrase I, II, IX, and XII Inhibition Studies with a Series of Cyclic Sulfonyl Guanidines","volume":"19","author":"Abdoli","year":"2024","journal-title":"ChemMedChem"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2335927","DOI":"10.1080\/14756366.2024.2335927","article-title":"Synthesis, carbonic anhydrase inhibition studies and modelling investigations of phthalimide-hydantoin hybrids","volume":"39","author":"Abdoli","year":"2024","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_31","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_32","doi-asserted-by":"crossref","first-page":"5528","DOI":"10.1021\/ja068359w","article-title":"Structural analysis of charge discrimination in the binding of inhibitors to human carbonic anhydrases I and II","volume":"129","author":"Srivastava","year":"2007","journal-title":"J. Am. Chem. Soc."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1107\/S0907444910006554","article-title":"Atomic resolution studies of carbonic anhydrase II","volume":"66","author":"Behnke","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"9004","DOI":"10.1021\/acs.jmedchem.5b01343","article-title":"Efficient Expression and Crystallization System of Cancer-Associated Carbonic Anhydrase Isoform IX","volume":"58","author":"Leitans","year":"2015","journal-title":"J. Med. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"9545","DOI":"10.1073\/pnas.161301298","article-title":"Crystal structure of the dimeric extracellular domain of human carbonic anhydrase XII, a bitopic membrane protein overexpressed in certain cancer tumor cells","volume":"98","author":"Whittington","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1080\/14756366.2017.1356295","article-title":"Synthesis and carbonic anhydrase I, II, VII, and IX inhibition studies with a series of benzo[d]thiazole-5- and 6-sulfonamides","volume":"32","author":"Abdoli","year":"2017","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Abdoli, M., Bozdag, M., Angeli, A., and Supuran, C.T. (2018). Benzamide-4-Sulfonamides Are Effective Human Carbonic Anhydrase I, II, VII, and IX Inhibitors. Metabolites, 8.","DOI":"10.3390\/metabo8020037"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1093\/nar\/28.1.235","article-title":"The Protein Data Bank","volume":"28","author":"Berman","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref_39","unstructured":"(Schr\u00f6dinger Suite Release, 2024). Schr\u00f6dinger Suite Release, version 2024-3; (a) Maestro v.14.1; (b) Epik, v.6.9; (c) Macromodel v.14.5; (d) Glide, v.10.4; (e) Desmond, v.7.9; (f) Prime, v.5.5; (g) Impact, v.10.4; (h) Jaguar, v.12.5."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4291","DOI":"10.1021\/acs.jctc.1c00302","article-title":"OPLS4: Improving Force Field Accuracy on Challenging Regimes of Chemical Space","volume":"17","author":"Lu","year":"2021","journal-title":"J. Chem. Theory Comput."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"6474","DOI":"10.1021\/jp003919d","article-title":"Evaluation and Reparametrization of the OPLS-AA Force Field for Proteins via Comparison with Accurate Quantum Chemical Calculations on Peptides","volume":"105","author":"Kaminski","year":"2001","journal-title":"J. Phys. Chem. B"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Angeli, A., Paoletti, N., and Supuran, C.T. (2023). Five-Membered Heterocyclic Sulfonamides as Carbonic Anhydrase Inhibitors. Molecules, 28.","DOI":"10.3390\/molecules28073220"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1605","DOI":"10.1002\/jcc.20084","article-title":"UCSF Chimera\u2014A visualization system for exploratory research and analysis","volume":"25","author":"Pettersen","year":"2004","journal-title":"J. Comput. Chem."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/1\/151\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T11:38:16Z","timestamp":1768477096000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/1\/151"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,14]]},"references-count":43,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["ph19010151"],"URL":"https:\/\/doi.org\/10.3390\/ph19010151","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,14]]}}}