{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T18:53:53Z","timestamp":1785437633877,"version":"3.56.0"},"posted":{"date-parts":[[2025,7,4]]},"group-title":"Pharmacology and Toxicology","reference-count":35,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,7,4]],"date-time":"2025-07-04T00:00:00Z","timestamp":1751587200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["1U19AI171292-01"],"award-info":[{"award-number":["1U19AI171292-01"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,7,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The emergence of mosquito-borne alphaviruses that cause chronic arthritis or encephalitis underscores the urgent need for broad-spectrum antiviral therapeutics. The viral nsP2 cysteine protease, which is essential for alphavirus replication, is a promising antiviral target. Vinyl sulfone-based inhibitors, such as RA-2034, potently inhibit nsP2 protease but suffer from glutathione reactivity and species-dependent systemic clearance catalyzed by glutathione\n                  <jats:italic>S<\/jats:italic>\n                  -transferase. To address these liabilities, we explored alternative electrophilic warheads and identified reverse amide inhibitors bearing\n                  <jats:italic>N<\/jats:italic>\n                  -alkyl sulfamate warheads with improved biochemical and antiviral profiles.\n                  <jats:italic>N<\/jats:italic>\n                  -methyl sulfamate acetamide\n                  <jats:bold>5<\/jats:bold>\n                  emerged as a lead compound with potency against both New and Old World alphaviruses, low GSH reactivity, and high proteome-wide selectivity. Despite its promising antialphaviral activity,\n                  <jats:bold>5<\/jats:bold>\n                  exhibited rapid clearance due to hepatic glucuronidation. Structure\u2013activity studies revealed modifications that improve metabolic stability while retaining antiviral activity. These findings introduce sulfamate acetamides as a new class of covalent nsP2 protease inhibitors and advance the discovery of direct acting pan-alphavirus drugs.\n                <\/jats:p>","DOI":"10.1101\/2025.06.30.662352","type":"posted-content","created":{"date-parts":[[2025,7,4]],"date-time":"2025-07-04T17:41:25Z","timestamp":1751650885000},"source":"Crossref","is-referenced-by-count":1,"title":["<i>N<\/i>\n                  -Alkyl Sulfamates as a New Class of nsP2 Cysteine Protease Inhibitors with Broad Spectrum Antialphaviral Activity"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9359-6074","authenticated-orcid":false,"given":"Anirban","family":"Ghoshal","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4713-2241","authenticated-orcid":false,"given":"John D.","family":"Sears","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8684-8755","authenticated-orcid":false,"given":"Mohammad Anwar","family":"Hossain","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9133-9274","authenticated-orcid":false,"given":"Edwin G.","family":"Tse","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3392-2848","authenticated-orcid":false,"given":"Stefanie","family":"Howell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-9719-8302","authenticated-orcid":false,"given":"Jane E.","family":"Burdick","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-2965-477X","authenticated-orcid":false,"given":"Noah L.","family":"Morales","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1333-5083","authenticated-orcid":false,"given":"Sabian A.","family":"Martinez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-6845-1284","authenticated-orcid":false,"given":"Isabella","family":"Law","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-3338-4867","authenticated-orcid":false,"given":"Zachary J.","family":"Streblow","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6828-2492","authenticated-orcid":false,"given":"Daniel N.","family":"Streblow","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1847-5090","authenticated-orcid":false,"given":"Rafael Miguez","family":"Cou\u00f1ago","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-6391-3268","authenticated-orcid":false,"given":"Nathaniel J.","family":"Moorman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5829-8477","authenticated-orcid":false,"given":"Mark T.","family":"Heise","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4181-8223","authenticated-orcid":false,"given":"Timothy M.","family":"Willson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025070714441841000_2025.06.30.662352v1.1","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms8081167"},{"key":"2025070714441841000_2025.06.30.662352v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.lana.2023.100673"},{"key":"2025070714441841000_2025.06.30.662352v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2013.07.012"},{"key":"2025070714441841000_2025.06.30.662352v1.4","doi-asserted-by":"publisher","DOI":"10.1186\/s12941-020-00360-4"},{"key":"2025070714441841000_2025.06.30.662352v1.5","doi-asserted-by":"crossref","unstructured":"Bassetto, M. ; Brancale, A. 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