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Chemical tools to facilitate inhibitor discovery as well as detection of target engagement by hit or lead compounds from high\u2010throughput screens are therefore in urgent need. We here report novel, selective activity\u2010based probes that enable detection of the SARS\u2010CoV\u20102 main protease. The probes are based on acyloxymethyl ketone reactive electrophiles combined with a peptide sequence including unnatural amino acids that targets the nonprimed site of the main protease substrate binding cleft. They are the first activity\u2010based probes for the main protease of coronaviruses and display target labeling within a human proteome without background. We expect that these reagents will be useful in the drug\u2010development pipeline, not only for the current SARS\u2010CoV\u20102, but also for other coronaviruses.<\/jats:p>","DOI":"10.1002\/cbic.202000371","type":"journal-article","created":{"date-parts":[[2020,7,27]],"date-time":"2020-07-27T14:35:24Z","timestamp":1595860524000},"page":"3383-3388","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":35,"title":["Peptidyl Acyloxymethyl Ketones as Activity\u2010Based Probes for the Main Protease of SARS\u2010CoV\u20102**"],"prefix":"10.1002","volume":"21","author":[{"given":"Merel A. 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