{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T22:00:54Z","timestamp":1790632854076,"version":"4.1.0"},"reference-count":32,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2016,2,10]],"date-time":"2016-02-10T00:00:00Z","timestamp":1455062400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Dengue virus (DENV) infection causes one of the most widespread mosquito-borne diseases in the world. Despite the great need, effective vaccines and practical antiviral therapies are still under development. Intracellular lipid levels are regulated by sterol regulatory elements-binding proteins (SREBPs), which are activated by serine protease, site 1 protease (S1P). Small compound PF-429242 is known as a S1P inhibitor and the antivirus effects have been reported in some viruses. In this study, we examined the anti-DENV effects of PF-429242 using all four serotypes of DENV by several primate-derived cell lines. Moreover, emergence of drug-resistant DENV mutants was assessed by sequential passages with the drug. DENV dependency on intracellular lipids during their infection was also evaluated by adding extracellular lipids. The addition of PF-429242 showed suppression of viral propagation in all DENV serotypes. We showed that drug-resistant DENV mutants are unlikely to emerge after five times sequential passages through treatment with  PF-429242. Although the levels of intracellular cholesterol and lipid droplets were reduced by  PF-429242, viral propagations were not recovered by addition of exogenous cholesterol or fatty acids, indicating that the reduction of LD and cholesterol caused by PF-429242 treatment is not related to its mechanism of action against DENV propagation. Our results suggest that PF-429242 is a promising candidate for an anti-DENV agent.<\/jats:p>","DOI":"10.3390\/v8020046","type":"journal-article","created":{"date-parts":[[2016,2,10]],"date-time":"2016-02-10T10:08:25Z","timestamp":1455098905000},"page":"46","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Suppressive Effects of the Site 1 Protease (S1P) Inhibitor, PF-429242, on Dengue Virus Propagation"],"prefix":"10.3390","volume":"8","author":[{"given":"Leo","family":"Uchida","sequence":"first","affiliation":[{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuzo","family":"Urata","sequence":"additional","affiliation":[{"name":"Department of Emerging Infectious Disease, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gianne","family":"Ulanday","sequence":"additional","affiliation":[{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"},{"name":"Graduate school of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuki","family":"Takamatsu","sequence":"additional","affiliation":[{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiro","family":"Yasuda","sequence":"additional","affiliation":[{"name":"Department of Emerging Infectious Disease, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"},{"name":"Leading graduate school program, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kouichi","family":"Morita","sequence":"additional","affiliation":[{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"},{"name":"Leading graduate school program, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daisuke","family":"Hayasaka","sequence":"additional","affiliation":[{"name":"Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"},{"name":"Leading graduate school program, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2016,2,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1038\/nature12060","article-title":"The global distribution and burden of dengue","volume":"496","author":"Bhatt","year":"2013","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2853","DOI":"10.4161\/21645515.2014.972131","article-title":"Assessment of bivalent and tetravalent dengue vaccine formulations in flavivirus-naive adults in mexico","volume":"10","author":"Dayan","year":"2014","journal-title":"Hum. Vaccin Immunother."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.1056\/NEJMoa1506223","article-title":"Efficacy and long-term safety of a dengue vaccine in regions of endemic disease","volume":"373","author":"Hadinegoro","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_4","first-page":"96","article-title":"Hemorrhagic fever in thailand; recent knowledge regarding etiology","volume":"20","author":"Halstead","year":"1967","journal-title":"Jpn. J. Med. Sci. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1086\/315215","article-title":"Dengue viremia titer, antibody response pattern, and virus serotype correlate with disease severity","volume":"181","author":"Vaughn","year":"2000","journal-title":"J. Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4557","DOI":"10.1128\/JVI.79.8.4557-4567.2005","article-title":"Heat shock protein 90 and heat shock protein 70 are components of dengue virus receptor complex in human cells","volume":"79","author":"Medina","year":"2005","journal-title":"J. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Samsa, M.M., Mondotte, J.A., Iglesias, N.G., Assuncao-Miranda, I., Barbosa-Lima, G., Da Poian, A.T., Bozza, P.T., and Gamarnik, A.V. (2009). Dengue virus capsid protein usurps lipid droplets for viral particle formation. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000632"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"6470","DOI":"10.1128\/JVI.00117-08","article-title":"Cholesterol effectively blocks entry of flavivirus","volume":"82","author":"Lee","year":"2008","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1159\/000321892","article-title":"Statins reduce dengue virus production via decreased virion assembly","volume":"54","author":"Castellanos","year":"2011","journal-title":"Intervirology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"4411","DOI":"10.1016\/j.bmcl.2007.06.031","article-title":"Aminopyrrolidineamide inhibitors of site-1 protease","volume":"17","author":"Hay","year":"2007","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1124\/jpet.108.139626","article-title":"Pharmacologic inhibition of site 1 protease activity inhibits sterol regulatory element-binding protein processing and reduces lipogenic enzyme gene expression and lipid synthesis in cultured cells and experimental animals","volume":"326","author":"Hawkins","year":"2008","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1125","DOI":"10.1172\/JCI0215593","article-title":"Srebps: Activators of the complete program of cholesterol and fatty acid synthesis in the liver","volume":"109","author":"Horton","year":"2002","journal-title":"J. Clin. Investig."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/S0092-8674(00)80213-5","article-title":"The srebp pathway: Regulation of cholesterol metabolism by proteolysis of a membrane-bound transcription factor","volume":"89","author":"Brown","year":"1997","journal-title":"Cell."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"11041","DOI":"10.1073\/pnas.96.20.11041","article-title":"A proteolytic pathway that controls the cholesterol content of membranes, cells, and blood","volume":"96","author":"Brown","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Olmstead, A.D., Knecht, W., Lazarov, I., Dixit, S.B., and Jean, F. (2012). Human subtilase ski-1\/s1p is a master regulator of the hcv lifecycle and a potential host cell target for developing indirect-acting antiviral agents. PLoS Pathog., 8.","DOI":"10.1371\/journal.ppat.1002468"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.antiviral.2012.05.006","article-title":"Ski-1\/s1p inhibition: A promising surrogate to statins to block hepatitis c virus replication","volume":"95","author":"Blanchet","year":"2012","journal-title":"Antivir. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.antiviral.2014.12.017","article-title":"Ski-1\/s1p inhibitor pf-429242 impairs the onset of hcv infection","volume":"115","author":"Blanchet","year":"2015","journal-title":"Antivir. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.virol.2011.11.008","article-title":"Evaluation of the anti-arenaviral activity of the subtilisin kexin isozyme-1\/site-1 protease inhibitor pf-429242","volume":"423","author":"Pasquato","year":"2012","journal-title":"Virology"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1128\/JVI.02019-10","article-title":"Antiviral activity of a small-molecule inhibitor of arenavirus glycoprotein processing by the cellular site 1 protease","volume":"85","author":"Urata","year":"2011","journal-title":"J. Virol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1006\/viro.1997.8500","article-title":"Construction of infectious cDNA clones for dengue 2 virus: Strain 16681 and its attenuated vaccine derivative, strain PDK-53","volume":"230","author":"Kinney","year":"1997","journal-title":"Virology"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1089\/adt.2009.0196","article-title":"Quantification of lipid droplets and associated proteins in cellular models of obesity via high-content\/high-throughput microscopy and automated image analysis","volume":"7","author":"McDonough","year":"2009","journal-title":"Assay Drug Dev. Technol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Uchida, L., Espada-Murao, L.A., Takamatsu, Y., Okamoto, K., Hayasaka, D., Yu, F., Nabeshima, T., Buerano, C.C., and Morita, K. (2014). The dengue virus conceals double-stranded rna in the intracellular membrane to escape from an interferon response. Sci. Rep., 4.","DOI":"10.1038\/srep07395"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"343","DOI":"10.7883\/yoken.JJID.2009.343","article-title":"Isolation and characterization of two phenotypically distinct dengue type-2 virus isolates from the same dengue hemorrhagic fever patient","volume":"62","author":"Kinoshita","year":"2009","journal-title":"Jpn. J. Infect. Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1099\/vir.0.037853-0","article-title":"Dengue virus strain den2 16681 utilizes a specific glycochain of syndecan-2 proteoglycan as a receptor","volume":"93","author":"Okamoto","year":"2012","journal-title":"J. Gen. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2096","DOI":"10.1128\/JVI.06796-11","article-title":"Dengue virus capsid protein binding to hepatic lipid droplets (ld) is potassium ion dependent and is mediated by ld surface proteins","volume":"86","author":"Carvalho","year":"2012","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1038\/ncb1631","article-title":"The lipid droplet is an important organelle for hepatitis c virus production","volume":"9","author":"Miyanari","year":"2007","journal-title":"Nat. Cell. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"37158","DOI":"10.1074\/jbc.M707329200","article-title":"The lipid droplet binding domain of hepatitis C virus core protein is a major determinant for efficient virus assembly","volume":"282","author":"Shavinskaya","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1042\/BJ20112219","article-title":"The disordered n-terminal region of dengue virus capsid protein contains a lipid-droplet-binding motif","volume":"444","author":"Martins","year":"2012","journal-title":"Biochem. J."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.virol.2009.03.025","article-title":"Cholesterol biosynthesis modulation regulates dengue viral replication","volume":"389","author":"Rothwell","year":"2009","journal-title":"Virology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"6782","DOI":"10.1128\/JVI.01757-09","article-title":"Rotaviruses associate with cellular lipid droplet components to replicate in viroplasms, and compounds disrupting or blocking lipid droplets inhibit viroplasm formation and viral replication","volume":"84","author":"Cheung","year":"2010","journal-title":"J. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"8422","DOI":"10.1128\/JVI.02601-05","article-title":"Reovirus outer capsid protein micro1 induces apoptosis and associates with lipid droplets, endoplasmic reticulum, and mitochondria","volume":"80","author":"Coffey","year":"2006","journal-title":"J. Virol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Vogt, D.A., Camus, G., Herker, E., Webster, B.R., Tsou, C.L., Greene, W.C., Yen, T.S., and Ott, M. (2013). Lipid droplet-binding protein tip47 regulates hepatitis c virus rna replication through interaction with the viral ns5a protein. PLoS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003302"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/8\/2\/46\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T19:19:04Z","timestamp":1760210344000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/8\/2\/46"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,2,10]]},"references-count":32,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2016,2]]}},"alternative-id":["v8020046"],"URL":"https:\/\/doi.org\/10.3390\/v8020046","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,2,10]]}}}