{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T21:23:37Z","timestamp":1767993817526,"version":"3.49.0"},"reference-count":26,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,9,16]],"date-time":"2021-09-16T00:00:00Z","timestamp":1631750400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003130","name":"Fonds Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["1S38718N"],"award-info":[{"award-number":["1S38718N"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["21632008"],"award-info":[{"award-number":["21632008"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81620108027"],"award-info":[{"award-number":["81620108027"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Human noroviruses (HuNoVs) are the most common cause of viral gastroenteritis resulting annually in ~219,000 deaths and a societal cost of ~USD 60 billion, and no antivirals or vaccines are available. Here, we assess the anti-norovirus activity of new peptidomimetic aldehydes related to the protease inhibitor rupintrivir. The early hit compound 4 inhibited the replication of murine norovirus (MNV) and the HuNoV GI.1 replicon in vitro (EC50 ~1 \u00b5M) and swiftly cleared the HuNoV GI.1 replicon from the cells. Compound 4 still inhibits the proteolytic activity. We selected a resistant GI.1 replicon, with a mutation (I109V) in a highly conserved region of the viral protease, conferring a low yield of resistance against compound 4 and rupintrivir. After testing new derivatives, compound 10d was the most potent (EC50 nanomolar range). Molecular docking indicated that the aldehyde group of compounds 4 and 10d bind with Cys139 in the HuNoV 3CL protease by a covalent linkage. Finally, compound 10d inhibited the replication of HuNoV GII.4 in infected zebrafish larvae, and PK studies in mice showed an adequate profile.<\/jats:p>","DOI":"10.3390\/v13091852","type":"journal-article","created":{"date-parts":[[2021,9,22]],"date-time":"2021-09-22T03:47:35Z","timestamp":1632282455000},"page":"1852","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["A Novel Class of Norovirus Inhibitors Targeting the Viral Protease with Potent Antiviral Activity In Vitro and In Vivo"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1632-3499","authenticated-orcid":false,"given":"Jana","family":"Van Dycke","sequence":"first","affiliation":[{"name":"Laboratory of Virology and Chemotherapy, KU Leuven\u2013Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8508-7130","authenticated-orcid":false,"given":"Wenhao","family":"Dai","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zoe","family":"Stylianidou","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Chemotherapy, KU Leuven\u2013Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jian","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, China"},{"name":"College of Pharmacy, Nanjing University of Chinese Medicine, 138 Xianlin Avenue, Qixia District, Nanjing 210023, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8360-9510","authenticated-orcid":false,"given":"Arno","family":"Cuvry","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Chemotherapy, KU Leuven\u2013Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emma","family":"Roux","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Chemotherapy, KU Leuven\u2013Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bingqian","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3838-310X","authenticated-orcid":false,"given":"Jasper","family":"Rymenants","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Chemotherapy, KU Leuven\u2013Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lindsey","family":"Bervoets","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Chemotherapy, KU Leuven\u2013Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9989-9567","authenticated-orcid":false,"given":"Peter","family":"de Witte","sequence":"additional","affiliation":[{"name":"Laboratory for Molecular Biodiscovery, KU Leuven\u2013Department of Pharmaceutical and Pharmacological Sciences, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hong","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0033-7514","authenticated-orcid":false,"given":"Johan","family":"Neyts","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Chemotherapy, KU Leuven\u2013Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8299-3787","authenticated-orcid":false,"given":"Joana","family":"Rocha-Pereira","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Chemotherapy, KU Leuven\u2013Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Bartsch, S.M., Lopman, B.A., Ozawa, S., Hall, A.J., and Lee, B.Y. 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