{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T23:52:48Z","timestamp":1784073168722,"version":"3.55.0"},"reference-count":34,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2019,10,31]],"date-time":"2019-10-31T00:00:00Z","timestamp":1572480000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key Research and Development Program of China (2016YFD0500407), the National Science Fund for Distinguished Young Scholars of China (31825025) and National Natural Science Foundation of China 31972721), and Fundamental Research Funds for the Cent","award":["2016YFD0500407, 31825025, 31972721, 2662018QD025"],"award-info":[{"award-number":["2016YFD0500407, 31825025, 31972721, 2662018QD025"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Flaviviruses, such as Zika virus (ZIKV), Japanese encephalitis virus (JEV), Dengue virus (DENV), and West Nile virus (WNV), are important arthropod-borne pathogens that present an immense global health problem. Their unpredictable disease severity, unusual clinical features, and severe neurological manifestations underscore an urgent need for antiviral interventions. Furin, a host proprotein convertase, is a key contender in processing flavivirus prM protein to M protein, turning the inert virus to an infectious particle. For this reason, the current study was planned to evaluate the antiviral activity of decanoyl-Arg-Val-Lys-Arg-chloromethylketone, a specific furin inhibitor, against flaviviruses, including ZIKV and JEV. Analysis of viral proteins revealed a significant increase in the prM\/E index of ZIKV or JEV in dec-RVKR-cmk-treated Vero cells compared to DMSO-treated control cells, indicating dec-RVKR-cmk inhibits prM cleavage. Plaque assay, qRT-PCR, and immunofluorescence assay revealed a strong antiviral activity of dec-RVKR-cmk against ZIKV and JEV in terms of the reduction in virus progeny titer and in viral RNA and protein production in both mammalian cells and mosquito cells. Time-of-drug addition assay revealed that the maximum reduction of virus titer was observed in post-infection treatment. Furthermore, our results showed that dec-RVKR-cmk exerts its inhibitory action on the virus release and next round infectivity but not on viral RNA replication. Taken together, our study highlights an interesting antiviral activity of dec-RVKR-cmk against flaviviruses.<\/jats:p>","DOI":"10.3390\/v11111011","type":"journal-article","created":{"date-parts":[[2019,10,31]],"date-time":"2019-10-31T12:38:23Z","timestamp":1572525503000},"page":"1011","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":39,"title":["Decanoyl-Arg-Val-Lys-Arg-Chloromethylketone: An Antiviral Compound That Acts against Flaviviruses through the Inhibition of Furin-Mediated prM Cleavage"],"prefix":"10.3390","volume":"11","author":[{"given":"Muhammad","family":"Imran","sequence":"first","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, Hubei, China"},{"name":"Key Laboratory of Preventive Veterinary Medicine in Hubei Province, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430070, Hubei, China"},{"name":"Department of Pathology, Faculty of Veterinary Science, University of Agriculture, Faisalabad 38040, Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad Kashif","family":"Saleemi","sequence":"additional","affiliation":[{"name":"Department of Pathology, Faculty of Veterinary Science, University of Agriculture, Faisalabad 38040, Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zheng","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, Hubei, China"},{"name":"Key Laboratory of Preventive Veterinary Medicine in Hubei Province, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong 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Degradation","volume":"92","author":"Panayiotou","year":"2018","journal-title":"J. Virol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1016\/S0140-6736(08)60238-X","article-title":"Pathogenic flaviviruses","volume":"371","author":"Gould","year":"2008","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1598","DOI":"10.1016\/j.celrep.2019.01.036","article-title":"Flavivirus NS1 Triggers Tissue-Specific Vascular Endothelial Dysfunction Reflecting Disease Tropism","volume":"26","author":"Glasner","year":"2019","journal-title":"Cell Rep."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Kim, S., Li, B., and Linhardt, R. (2017). Pathogenesis and inhibition of flaviviruses from a carbohydrate perspective. Pharmaceuticals, 10.","DOI":"10.3390\/ph10020044"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"4301","DOI":"10.1016\/j.vaccine.2011.09.114","article-title":"Flaviviruses and flavivirus vaccines","volume":"30","author":"Heinz","year":"2012","journal-title":"Vaccine"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"17006","DOI":"10.1038\/celldisc.2017.6","article-title":"Zika virus evades interferon-mediated antiviral response through the co-operation of multiple nonstructural proteins in vitro","volume":"3","author":"Wu","year":"2017","journal-title":"Cell Discov."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1797","DOI":"10.3389\/fmicb.2018.01797","article-title":"Role of Zika Virus prM protein in viral pathogenicity and use in vaccine development","volume":"9","author":"Nambala","year":"2018","journal-title":"Front. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"4370","DOI":"10.1128\/JVI.77.7.4370-4382.2003","article-title":"Intracellular assembly and secretion of recombinant subviral particles from tick-borne encephalitis virus","volume":"77","author":"Lorenz","year":"2003","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"10787","DOI":"10.1128\/JVI.75.22.10787-10799.2001","article-title":"Assembly and maturation of the flavivirus Kunjin virus appear to occur in the rough endoplasmic reticulum and along the secretory pathway, respectively","volume":"75","author":"Mackenzie","year":"2001","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1830","DOI":"10.1126\/science.1153263","article-title":"The flavivirus precursor membrane-envelope protein complex: Structure and maturation","volume":"319","author":"Li","year":"2008","journal-title":"Science"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1834","DOI":"10.1126\/science.1153264","article-title":"Structure of the immature dengue virus at low pH primes proteolytic maturation","volume":"319","author":"Yu","year":"2008","journal-title":"Science"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"11813","DOI":"10.1128\/JVI.79.18.11813-11823.2005","article-title":"Resuscitating mutations in a furin cleavage-deficient mutant of the flavivirus tick-borne encephalitis virus","volume":"79","author":"Elshuber","year":"2005","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1099\/vir.0.035964-0","article-title":"A conserved region in the prM protein is a critical determinant in the assembly of flavivirus particles","volume":"93","author":"Yoshii","year":"2012","journal-title":"J. Gen. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1016\/0300-9084(94)90149-X","article-title":"Processing of viral glycoproteins by the subtilisin-like endoprotease furin and its inhibition by specific peptidylchloroalkylketones","volume":"76","author":"Garten","year":"1994","journal-title":"Biochimie"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1067","DOI":"10.1021\/jm9012455","article-title":"Potent inhibitors of furin and furin-like proprotein convertases containing decarboxylated P1 arginine mimetics","volume":"53","author":"Becker","year":"2010","journal-title":"J. Med. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1230","DOI":"10.1111\/liv.12185","article-title":"Therapeutic potential of furin inhibitors for the chronic infection of hepatitis B virus","volume":"33","author":"Pang","year":"2013","journal-title":"Liver Int."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1038\/360358a0","article-title":"Inhibition of furin-mediated cleavage activation of HIV-1 glycoprotein gpl60","volume":"360","author":"Hallenberger","year":"1992","journal-title":"Nature"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"21899","DOI":"10.1074\/jbc.M802444200","article-title":"Inhibition of Chikungunya virus infection in cultured human muscle cells by furin inhibitors impairment of the maturation of the E2 surface glycoprotein","volume":"283","author":"Ozden","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4569","DOI":"10.1128\/JVI.02281-09","article-title":"Initiation of duck hepatitis B virus infection requires cleavage by a furin-like protease","volume":"84","author":"Tong","year":"2010","journal-title":"J. Virol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.2217\/fmb.09.86","article-title":"The role of furin in papillomavirus infection","volume":"4","author":"Day","year":"2009","journal-title":"Future Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1517\/13543776.2014.1000303","article-title":"Therapeutic uses of furin and its inhibitors: A patent review","volume":"25","author":"Couture","year":"2015","journal-title":"Expert Opin. Ther. Pat."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4960","DOI":"10.2741\/3055","article-title":"Knock-out mouse models of proprotein convertases: Unique functions or redundancy?","volume":"13","author":"Creemers","year":"2008","journal-title":"Front. Biosci. J. Virtual Libr."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1515\/BMC.2011.034","article-title":"On the cutting edge of proprotein convertase pharmacology: From molecular concepts to clinical applications","volume":"2","author":"Couture","year":"2011","journal-title":"Biomol. Concepts"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"53442","DOI":"10.1074\/jbc.M407152200","article-title":"Limited redundancy of the proprotein convertase furin in mouse liver","volume":"279","author":"Roebroek","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3722","DOI":"10.1128\/JVI.03242-15","article-title":"MicroRNA-33a-5p modulates Japanese Encephalitis Virus replication by targeting eukaryotic translation elongation factor 1A1","volume":"90","author":"Chen","year":"2016","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"12067","DOI":"10.1128\/JVI.05559-11","article-title":"The infectivity of prM-containing partially mature West Nile virus does not require the activity of cellular furin-like proteases","volume":"85","author":"Mukherjee","year":"2011","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Kovanich, D., Saisawang, C., Sittipaisankul, P., Ramphan, S., Kalpongnukul, N., Somparn, P., Pisitkun, T., and Smith, D.R. (2019). Analysis of the Zika and Japanese encephalitis virus NS5 interactomes. J. Proteome Res.","DOI":"10.1021\/acs.jproteome.9b00318"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"21992","DOI":"10.1074\/jbc.M111.332643","article-title":"Highly potent inhibitors of proprotein convertase furin as potential drugs for treatment of infectious diseases","volume":"287","author":"Becker","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1016\/j.antiviral.2017.03.026","article-title":"Luteolin restricts dengue virus replication through inhibition of the proprotein convertase furin","volume":"143","author":"Peng","year":"2017","journal-title":"Antivir. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1038\/nrm934","article-title":"Furin at the cutting edge: From protein traffic to embryogenesis and disease","volume":"3","author":"Thomas","year":"2002","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"12240","DOI":"10.1016\/S0021-9258(17)32707-2","article-title":"The convertases furin and PC1 can both cleave the human immunodeficiency virus (HIV)-1 envelope glycoprotein gp160 into gp120 (HIV-1 SU) and gp41 (HIV-I TM)","volume":"269","author":"Decroly","year":"1994","journal-title":"J. Biol. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/S0168-1702(00)00149-0","article-title":"Cleavage of the respiratory syncytial virus fusion protein is required for its surface expression: Role of furin","volume":"68","author":"Bolt","year":"2000","journal-title":"Virus Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/S0962-8924(98)01382-8","article-title":"Bi-cycling the furin pathway: From TGN localization to pathogen activation and embryogenesis","volume":"9","author":"Molloy","year":"1999","journal-title":"Trends Cell Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1016\/j.virol.2010.12.051","article-title":"Hepatitis C virus-induced furin and thrombospondin-1 activate TGF-beta1: Role of TGF-beta1 in HCV replication","volume":"412","author":"Presser","year":"2011","journal-title":"Virology"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/11\/11\/1011\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:30:59Z","timestamp":1760189459000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/11\/11\/1011"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,10,31]]},"references-count":34,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2019,11]]}},"alternative-id":["v11111011"],"URL":"https:\/\/doi.org\/10.3390\/v11111011","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,10,31]]}}}