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Herein, we show that ligand screening can be performed in living human cells by \u201cintracellular protein\u2010observed\u201d NMR spectroscopy, without requiring enzymatic activity measurements or other cellular assays. Quantitative binding information is obtained by fast, inexpensive <jats:sup>1<\/jats:sup>H\u2005NMR experiments, providing intracellular dose\u2010 and time\u2010dependent ligand binding curves, from which kinetic and thermodynamic parameters linked to cell permeability and binding affinity and selectivity are obtained. The approach was applied to carbonic anhydrase and, in principle, can be extended to any NMR\u2010observable intracellular target. The results obtained are directly related to the potency of candidate drugs, that is, the required dose. The application of this approach at an early stage of the drug design pipeline could greatly increase the low success rate of modern drug development.<\/jats:p>","DOI":"10.1002\/anie.201913436","type":"journal-article","created":{"date-parts":[[2020,2,5]],"date-time":"2020-02-05T10:16:49Z","timestamp":1580897809000},"page":"6535-6539","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":66,"title":["Drug Screening in Human Cells by NMR Spectroscopy Allows the Early Assessment of Drug Potency"],"prefix":"10.1002","volume":"59","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4183-4311","authenticated-orcid":false,"given":"Enrico","family":"Luchinat","sequence":"first","affiliation":[{"name":"CERM\u2014Magnetic Resonance Center Universit\u00e0 degli Studi di Firenze  via Luigi Sacconi 6 50019 Sesto Fiorentino Italy"},{"name":"Dipartimento di Scienze Biomediche Sperimentali e Cliniche \u201cMario Serio\u201d Universit\u00e0 degli 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