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Here, we report a high\u2010throughput screening for anti\u2010smallpox virus drugs from a 767\u2010small\u2010molecule library, employing two vaccinia virus (VACV) strains containing firefly luciferase (VTT\u2010Fluc and VG9\u2010Fluc) as surrogate viruses. Using an eight\u2010point dose response format assay, 26 compounds of different pharmacological classes were identified with in vitro anti\u2010VACV activities. Mycophenolate mofetil (MMF) and tranilast (TRA) were detected to possess the highest anti\u2010VACV potency (selectivity index values of &gt;334 and &gt;74, respectively); they could inhibit VTT\u2010Fluc replication in nude mice at 5 days post\u2010infection by 99% (10\u2009mg\/kg, <jats:italic>P<\/jats:italic>\u2009&lt;\u2009.01) and 59% (45\u2009mg\/kg, <jats:italic>P<\/jats:italic>\u2009=\u2009.01), respectively, as indicated by bioluminescent intensity. In conclusion, MMF and TRA are promising anti\u2010smallpox virus candidates for further optimization and repurposing for use in clinical practice.<\/jats:p>","DOI":"10.1002\/jmv.25544","type":"journal-article","created":{"date-parts":[[2019,7,11]],"date-time":"2019-07-11T14:26:01Z","timestamp":1562855161000},"page":"2016-2024","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Screening and evaluation of potential inhibitors against vaccinia virus from 767 approved drugs"],"prefix":"10.1002","volume":"91","author":[{"given":"Jiajing","family":"Wu","sequence":"first","affiliation":[{"name":"National Engineering Technology Research Center of Combination Vaccines Wuhan Institute of Biological Products Wuhan Hubei China"},{"name":"Division of HIV\/AIDS and Sex\u2010transmitted Virus Vaccines National Institutes for Food and Drug Control Beijing 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