{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T03:02:38Z","timestamp":1789959758578,"version":"4.0.1"},"reference-count":28,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,3,22]],"date-time":"2026-03-22T00:00:00Z","timestamp":1774137600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,3,22]],"date-time":"2026-03-22T00:00:00Z","timestamp":1774137600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100000780","name":"European Commission","doi-asserted-by":"publisher","award":["101094131"],"award-info":[{"award-number":["101094131"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Magnetic Reson in Chemistry"],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Drug development is a risky endeavour with a high failure rate, often caused by the limited ability to predict the efficacy and interactions of candidate drugs in a native cellular environment. In this context, in\u2010cell NMR spectroscopy is a promising tool for assessing drug\u2010target binding directly in living cells, thereby improving the screening and development of new molecules. In this study, we used real\u2010time in\u2010cell\n                    <jats:sup>19<\/jats:sup>\n                    F NMR spectroscopy in a flow bioreactor to observe competitive binding of fluorinated benzenesulfonamide derivatives to three cytosolic isoforms of carbonic anhydrase. Quantitative measurement of the dissociation constants relative to a spy ligand allowed an accurate ranking of the compounds based on their intracellular affinities for each isoform. The use of two fluorinated ligands allowed simultaneous observation of spy ligand displacement and test ligand binding, as well as estimation of the effective ratio of free ligand concentrations under poor solubility conditions. We also show that signal saturation caused by short repetition times, which can significantly impact the analysis, can be easily corrected a posteriori. Overall, we show that real\u2010time in\u2010cell\n                    <jats:sup>19<\/jats:sup>\n                    F NMR spectroscopy can reliably quantify drug\u2013target binding in the cellular environment, paving the way for future applications in drug discovery.\n                  <\/jats:p>","DOI":"10.1002\/mrc.70098","type":"journal-article","created":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T06:24:52Z","timestamp":1774247092000},"page":"584-593","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Quantitative Competition Binding of Fluorinated Ligands by Real\u2010Time\n                    <sup>19<\/sup>\n                    F In\u2010Cell NMR to Assess Isoform Selectivity in Human Cells"],"prefix":"10.1002","volume":"64","author":[{"given":"Azzurra","family":"Costantino","sequence":"first","affiliation":[{"name":"Magnetic Resonance Center\u2014CERM University of Florence  Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7457-490X","authenticated-orcid":false,"given":"Letizia","family":"Barbieri","sequence":"additional","affiliation":[{"name":"Magnetic Resonance Center\u2014CERM University of Florence  Sesto Fiorentino Italy"},{"name":"Interuniversity Consortium for Magnetic Resonance of Metalloproteins\u2014CIRMMP  Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0669-7481","authenticated-orcid":false,"given":"Simone","family":"Giovannuzzi","sequence":"additional","affiliation":[{"name":"NEUROFARBA Department, Section of Pharmaceutical and Nutraceutical Sciences University of Florence  Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3342-702X","authenticated-orcid":false,"given":"Alessio","family":"Nocentini","sequence":"additional","affiliation":[{"name":"NEUROFARBA Department, Section of Pharmaceutical and Nutraceutical Sciences University of Florence  Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4262-0323","authenticated-orcid":false,"given":"Claudiu\u00a0T.","family":"Supuran","sequence":"additional","affiliation":[{"name":"NEUROFARBA Department, Section of Pharmaceutical and Nutraceutical Sciences University of Florence  Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4183-4311","authenticated-orcid":false,"given":"Enrico","family":"Luchinat","sequence":"additional","affiliation":[{"name":"Magnetic Resonance Center\u2014CERM University of Florence  Sesto Fiorentino Italy"},{"name":"Interuniversity Consortium for Magnetic Resonance of Metalloproteins\u2014CIRMMP  Sesto Fiorentino Italy"},{"name":"Chemistry Department University of Florence  Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,3,22]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.apsb.2022.02.002"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrd4609"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmr.2019.07.018"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2022.102374"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pnmrs.2022.04.002"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.9b03031"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201913436"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.accounts.9b00377"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1039\/D1CB00085C"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10858-020-00325-x"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202201097"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.2c12086"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/chem.202303988"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jmedchem.3c01600"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pnmrs.2023.07.001"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja020174b"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmr.2009.10.008"},{"key":"e_1_2_9_19_1","first-page":"305","volume-title":"Methods Enzymol","author":"Burz D. 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