Discovery and Characterization of BAY-805, a Potent and Selective Inhibitor of Ubiquitin-Specific Protease USP21

• Published in: Journal of medicinal chemistry Vol. 66; no. 5; pp. 3431 - 3447
• Main Authors: Göricke, Fabian, Vu, Victoria, Smith, Leanna, Scheib, Ulrike, Böhm, Raphael, Akkilic, Namik, Wohlfahrt, Gerd, Weiske, Jörg, Bömer, Ulf, Brzezinka, Krzysztof, Lindner, Niels, Lienau, Philip, Gradl, Stefan, Beck, Hartmut, Brown, Peter J., Santhakumar, Vijayaratnam, Vedadi, Masoud, Barsyte-Lovejoy, Dalia, Arrowsmith, Cheryl H., Schmees, Norbert, Petersen, Kirstin
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Chemical Society 09.03.2023; Amer Chemical Soc
• Subjects: Chemistry, Medicinal; Endopeptidases; Gene Expression Regulation; Life Sciences & Biomedicine; Pharmacology & Pharmacy; Science & Technology; Signal Transduction; Ubiquitin-Specific Proteases; STRUCTURE-GUIDED DEVELOPMENT; CELL-CYCLE; PROLIFERATION; ASSAY; DEUBIQUITINATION
• ISSN: 0022-2623; 1520-4804
• DOI: 10.1021/acs.jmedchem.2c01933

USP21 belongs to the ubiquitin-specific protease (USP) subfamily of deubiquitinating enzymes (DUBs). Due to its relevance in tumor development and growth, USP21 has been reported as a promising novel therapeutic target for cancer treatment. Herein, we present the discovery of the first highly potent and selective USP21 inhibitor. Following high-throughput screening and subsequent structure-based optimization, we identified BAY-805 to be a non-covalent inhibitor with low nanomolar affinity for USP21 and high selectivity over other DUB targets as well as kinases, proteases, and other common off-targets. Furthermore, surface plasmon resonance (SPR) and cellular thermal shift assays (CETSA) demonstrated high-affinity target engagement of BAY-805, resulting in strong NF-κB activation in a cell-based reporter assay. To the best of our knowledge, BAY-805 is the first potent and selective USP21 inhibitor and represents a valuable high-quality in vitro chemical probe to further explore the complex biology of USP21.