biostudies-literatureAltincekic NHessisches Ministerium f?r Wissenschaft und KunstIntramural NIH HHSAgenzia Nazionale di Valutazione del Sistema Universitario e della RicercaSwiss National Science FoundationDeutsche ForschungsgemeinschaftNational Institute of Diabetes and Digestive and Kidney DiseasesH2020 Research InfrastructuresKnut och Alice Wallenbergs StiftelseGoethe-Universit?t Frankfurt am MainKrebsliga Schweiz563-574https://www.ebi.ac.uk/biostudies/studies/S-EPMC10877576biostudies-literatureUnknown19(2)The main protease M^pro, nsp5, of SARS-CoV-2 (SCoV2) is one of its most attractive drug targets. Here, we report primary screening data using nuclear magnetic resonance spectroscopy (NMR) of four different libraries and detailed follow-up synthesis on the promising uracil-containing fragment Z604 derived from these libraries. Z604 shows time-dependent binding. Its inhibitory effect is sensitive to reducing conditions. Starting with Z604, we synthesized and characterized 13 compounds designed by fragment growth strategies. Each compound was characterized by NMR and/or activity assays to investigate their interaction with M^pro. These investigations resulted in the four-armed compound <b>35b</b> that binds directly to M^pro. <b>35b</b> could be cocrystallized wiACS chemical biologyTargeting the Main Protease (M&lt;sup&gt;pro&lt;/sup&gt;, nsp5) by Growth of Fragment Scaffolds Exploiting Structure-Based Methodologies.PMC10877576NIH DK029046452632086277478796310030KFS-4903-08-201987103770653611CRC902101094131IWB-EFRE- 20007375192646310030_1926463926823092020.0182277479031Z01 DK029046Langer JAltincekic NLinhard VPerrakis AFragai MRichter CKarlsson BGCantini FBax AHedenstrom MSauer UHBarthel TWeiss MSBanci LAbi Saad MJButikofer MKaderli JJores NBerg HOrts JGrobner GBrath UBlommers MJJLohr FGande SLWollenhaupt JRobertson AOzturk SSchwalbe HSreeramulu SGrifagni DEhrhardt CfalseTargeting the Main Protease (M&lt;sup&gt;pro&lt;/sup&gt;, nsp5) by Growth of Fragment Scaffolds Exploiting Structure-Based Methodologies.The main protease M^pro, nsp5, of SARS-CoV-2 (SCoV2) is one of its most attractive drug targets. Here, we report primary screening data using nuclear magnetic resonance spectroscopy (NMR) of four different libraries and detailed follow-up synthesis on the promising uracil-containing fragment Z604 derived from these libraries. Z604 shows time-dependent binding. Its inhibitory effect is sensitive to reducing conditions. Starting with Z604, we synthesized and characterized 13 compounds designed by fragment growth strategies. Each compound was characterized by NMR and/or activity assays to investigate their interaction with M^pro. These investigations resulted in the four-armed compound <b>35b</b> that binds directly to M^pro. <b>35b</b> could be cocrystallized wi2024-01-01T00:00:00Z2024 Feb2026-06-01T23:46:05.626Z2025-04-19T07:35:42.022ZS-EPMC108775763823296010.1021/acschembio.3c00720