{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T15:51:30Z","timestamp":1792511490432,"version":"4.5.0"},"reference-count":27,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2021,7,16]],"date-time":"2021-07-16T00:00:00Z","timestamp":1626393600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004781","name":"Institut Pengurusan dan Pemantauan Penyelidikan, Universiti Malaya","doi-asserted-by":"publisher","award":["TR001B-2014B"],"award-info":[{"award-number":["TR001B-2014B"]}],"id":[{"id":"10.13039\/501100004781","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Zika virus (ZIKV) represents a re-emerging threat to global health due to its association with congenital birth defects. ZIKV NS2B-NS3 protease is crucial for virus replication by cleaving viral polyprotein at various junctions to release viral proteins and cause cytotoxic effects in ZIKV-infected cells. This study characterized the inhibitory effects of doxycycline against ZIKV NS2B-NS3 protease and viral replication in human skin cells. The in silico data showed that doxycycline binds to the active site of ZIKV protease at a low docking energy (\u22127.8 Kcal\/mol) via four hydrogen bonds with the protease residues TYR1130, SER1135, GLY1151, and ASP83. Doxycycline efficiently inhibited viral NS2B-NS3 protease at average human temperature (37 \u00b0C) and human temperature with a high fever during virus infection (40 \u00b0C). Interestingly, doxycycline showed a higher inhibitory effect at 40 \u00b0C (IC50 = 5.3 \u00b5M) compared to 37 \u00b0C (9.9 \u00b5M). The virus replication was considerably reduced by increasing the concentration of doxycycline. An approximately 50% reduction in virus replication was observed at 20 \u00b5M of doxycycline. Treatment with 20 \u00b5M of doxycycline reduced the cytopathic effects (CPE), and the 40 \u00b5M of doxycycline almost eliminated the CPE of human skin cells. This study showed that doxycycline binds to the ZIKV protease and inhibits its catalytic activity at a low micro-molecular concentration range. Treatment of human skin fibroblast with doxycycline eliminated ZIKV infection and protected the cells against the cytopathic effects of the infection.<\/jats:p>","DOI":"10.3390\/molecules26144321","type":"journal-article","created":{"date-parts":[[2021,7,18]],"date-time":"2021-07-18T21:16:48Z","timestamp":1626643008000},"page":"4321","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Doxycycline Interferes with Zika Virus Serine Protease and Inhibits Virus Replication in Human Skin Fibroblasts"],"prefix":"10.3390","volume":"26","author":[{"given":"Teow","family":"Chong Teoh","sequence":"first","affiliation":[{"name":"Bioinformatics Programme, Institute of Biological Sciences, Faculty of Science, Kuala Lumpur 50603, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sawsam","family":"J. Al-Harbi","sequence":"additional","affiliation":[{"name":"Department of Anatomy, College of Medicine, University of Babylon, Babylon, Iraq"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ammar Yasir","family":"Abdulrahman","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7300-6388","authenticated-orcid":false,"given":"Hussin A.","family":"Rothan","sequence":"additional","affiliation":[{"name":"Department of Biology, College of Arts and Sciences, Georgia State University, Atlanta, GA 30303, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,7,16]]},"reference":[{"key":"ref_1","first-page":"294","article-title":"Zika Virus","volume":"375","author":"Petersen","year":"2016","journal-title":"N. Engl. J. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/0035-9203(52)90043-6","article-title":"Zika virus. II. Pathogenicity and physical properties","volume":"46","author":"Dick","year":"1952","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.jaut.2018.01.002","article-title":"Current concerns and perspectives on Zika virus co-infection with arboviruses and HIV","volume":"89","author":"Rothan","year":"2018","journal-title":"J. Autoimmun."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1016\/0035-9203(64)90201-9","article-title":"Zika Virus Infection in Man","volume":"58","author":"Simpson","year":"1964","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1232","DOI":"10.3201\/eid1408.080287","article-title":"Genetic and serologic properties of Zika virus associated with an epidemic, Yap State, Micronesia, 2007","volume":"14","author":"Lanciotti","year":"2008","journal-title":"Emerg. Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1007\/s00705-006-0903-z","article-title":"Full-length sequencing and genomic characterization of Bagaza, Kedougou, and Zika viruses","volume":"152","author":"Kuno","year":"2007","journal-title":"Arch. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.gene.2017.07.049","article-title":"Molecular cloning and characterization of the genes encoding the proteins of Zika virus","volume":"628","author":"Hou","year":"2017","journal-title":"Gene"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"251482","DOI":"10.1155\/2012\/251482","article-title":"Protegrin-1 inhibits dengue NS2B-NS3 serine protease and viral replication in MK2 cells","volume":"2012","author":"Rothan","year":"2012","journal-title":"J. Biomed. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Rothan, H.A., Han, H.C., Ramasamy, T.S., Othman, S., Rahman, N.A., and Yusof, R. (2012). Inhibition of dengue NS2B-NS3 protease and viral replication in Vero cells by recombinant retrocyclin-1. BMC Infect. Dis., 12.","DOI":"10.1186\/1471-2334-12-314"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.virusres.2006.01.021","article-title":"Functional characterization of the NS2B\/NS3 protease complex from seven viruses belonging to different groups inside the genus Flavivirus","volume":"120","author":"Bessaud","year":"2006","journal-title":"Virus Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1007\/s00705-013-1880-7","article-title":"Inhibitory effect of doxycycline against dengue virus replication in vitro","volume":"159","author":"Rothan","year":"2014","journal-title":"Arch. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"8880","DOI":"10.1128\/JVI.00354-15","article-title":"Biology of Zika Virus Infection in Human Skin Cells","volume":"89","author":"Hamel","year":"2015","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1913","DOI":"10.1099\/0022-1317-69-8-1913","article-title":"Detection of virus-specific antigen in the nuclei or nucleoli of cells infected with Zika or Langat virus","volume":"69","author":"Buckley","year":"1988","journal-title":"J. Gen. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Rothan, H.A., and Kumar, M. (2019). Role of Endoplasmic Reticulum-Associated Proteins in Flavivirus Replication and Assembly Complexes. Pathogens, 8.","DOI":"10.3390\/pathogens8030148"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1016\/j.neuron.2019.01.010","article-title":"Zika Virus Protease Cleavage of Host Protein Septin-2 Mediates Mitotic Defects in Neural Progenitors","volume":"101","author":"Li","year":"2019","journal-title":"Neuron."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"45762","DOI":"10.1074\/jbc.M107360200","article-title":"Activity of recombinant dengue 2 virus NS3 protease in the presence of a truncated NS2B co-factor, small peptide substrates, and inhibitors","volume":"276","author":"Leung","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_17","first-page":"681","article-title":"Study the antiviral activity of some derivatives of tetracycline and non-steroid anti-inflammatory drugs towards dengue virus","volume":"30","author":"Rothan","year":"2013","journal-title":"Trop. Biomed."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/j.apjtm.2016.01.033","article-title":"Pharmacodynamics of aminoglycosides and tetracycline derivatives against Japanese encephalitis virus","volume":"9","author":"Topno","year":"2016","journal-title":"Asian Pac. J. Trop. Med."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Rothan, H.A., Bahrani, H., Mohamed, Z., and Teoh, T.C. (2015). A combination of doxycycline and ribavirin alleviated chikungunya infection. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0126360"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1358\/mf.1998.20.8.487489","article-title":"Antimurine retroviral effect of doxycycline","volume":"20","author":"Sturtz","year":"1998","journal-title":"Methods Find. Exp. Clin. Pharmacol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Wu, Z.C., Wang, X., Wei, J.-C., Li, B.-B., Shao, D.-H., Li, Y.-M., Liu, K., Shi, Y.-Y., Zhou, B., and Qiu, Y.-F. (2015). Antiviral activity of doxycycline against vesicular stomatitis virus in vitro. FEMS Microbiol. Lett., 362.","DOI":"10.1093\/femsle\/fnv195"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1002\/prot.10104","article-title":"Increasing the precision of comparative models with YASARA NOVA--a self-parameterizing force field","volume":"47","author":"Krieger","year":"2002","journal-title":"Proteins."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1038\/nprot.2010.32","article-title":"The HADDOCK web server for data-driven biomolecular docking","volume":"5","author":"Bonvin","year":"2010","journal-title":"Nat. Protoc."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1784","DOI":"10.1074\/mcp.M000051-MCP201","article-title":"Building macromolecular assemblies by information-driven docking: Introducing the HADDOCK multibody docking server","volume":"9","author":"Karaca","year":"2010","journal-title":"Mol. Cell Proteomics."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1659","DOI":"10.1007\/s10989-019-09808-4","article-title":"Novel Peptides Inhibit Zika NS2B-NS3 Serine Protease and Virus Replication in Human Hepatic Cell Line","volume":"25","author":"Abdulrahman","year":"2019","journal-title":"Int. J. Pept. Res. Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"36","DOI":"10.47665\/tb.38.1.007","article-title":"Evaluation of neutralizing antibodies produced by papaya mosaic virus nanoparticles fused to the E2EP3 peptide epitope of Chikungunya envelope","volume":"38","author":"Teoh","year":"2021","journal-title":"Trop. Biomed."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"713","DOI":"10.47665\/tb.37.3.713","article-title":"Antiviral and virucidal activities of sulphated polysaccharides against Japanese encephalitis virus","volume":"37","author":"Yusof","year":"2020","journal-title":"Trop. Biomed."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/14\/4321\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:31:03Z","timestamp":1760164263000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/14\/4321"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,16]]},"references-count":27,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2021,7]]}},"alternative-id":["molecules26144321"],"URL":"https:\/\/doi.org\/10.3390\/molecules26144321","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,16]]}}}