{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T14:11:05Z","timestamp":1788876665822,"version":"build-2803163510"},"reference-count":26,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T00:00:00Z","timestamp":1788825600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP26fk0210198"],"award-info":[{"award-number":["JP26fk0210198"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP25fk0210132"],"award-info":[{"award-number":["JP25fk0210132"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"award":["JP26fk0210198"],"award-info":[{"award-number":["JP26fk0210198"]}],"id":[{"id":"https:\/\/ror.org\/004rtk039","id-type":"ROR","asserted-by":"publisher"}]},{"award":["JP25fk0210132"],"award-info":[{"award-number":["JP25fk0210132"]}],"id":[{"id":"https:\/\/ror.org\/004rtk039","id-type":"ROR","asserted-by":"publisher"}]},{"name":"University of Health Sciences and Pharmacy","award":["FRIF-20-134"],"award-info":[{"award-number":["FRIF-20-134"]}]},{"award":["FRIF-20-134"],"award-info":[{"award-number":["FRIF-20-134"]}],"id":[{"id":"https:\/\/ror.org\/01btzz102","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Hepatitis A virus (HAV) infection remains a global public health concern in both developing and developed countries. In the present study, we identified anti-HAV drugs, using AutoDock Vina Modeling software, and evaluated the compounds in vitro. Following cytotoxicity for Huh7 cells, 5 out of 10 compounds were selected. We evaluated effective HAV 3C protease inhibitors with activity against both HAV genotype IB HM175\/18f and HAV genotype IIIA HA11-1299-infected human hepatoma cells. Among the five compounds, we identified only one (KCN-A-12), which had an inhibitory effect on both HAV genotype IB HM175\/18f and HAV genotype IIIA HA11-1299 replication in human hepatoma Huh7 cells. KCN-A-12 has more effective inhibitory effects on the replication of HAV genotype IB HM175\/18f than that of HAV genotype IIIA HA11-1299. This difference may be attributable to the fact that our discovery system depends on crystal structures from HAV 3C protease based on the HAV genotype IB HM175 strain. In conclusion, we observed that KCN-A-12 was able to inhibit HAV replication. In silico screening for HAV 3C protease inhibitors may be useful for further discovery of anti-HAV drugs.<\/jats:p>","DOI":"10.3390\/microorganisms14091987","type":"journal-article","created":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T13:46:33Z","timestamp":1788875193000},"page":"1987","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Targeting the 3C Protease of Hepatitis A Virus Subgenotype IB: Virtual Screening and Identification of Potent Lead Candidates"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-8135-342X","authenticated-orcid":false,"given":"Tatsuo","family":"Kanda","sequence":"first","affiliation":[{"name":"Division of Gastroenterology and Hepatology, Uonuma Institute of Community Medicine, Niigata University Medical and Dental Hospital, 4132 Urasa, Minamiuonuma 949-7302, Japan"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"given":"Reina","family":"Sasaki-Tanaka","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology and Hepatology, Niigata University Graduate School of Medicine, Dentistry and Health Sciences (Medicine), Niigata University, Niigata 951-8510, Japan"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0827-0964","authenticated-orcid":false,"given":"Hiroaki","family":"Okamoto","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, Shimotsuke 329-0498, Japan"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"supervision","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5439-635X","authenticated-orcid":false,"given":"Shuji","family":"Terai","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology and Hepatology, Niigata University Graduate School of Medicine, Dentistry and Health Sciences (Medicine), Niigata University, Niigata 951-8510, Japan"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"supervision","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Cole D.","family":"Cwiklowski","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, University of Health Sciences and Pharmacy, St. Louis, MO 63010, USA"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"resources","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7308-014X","authenticated-orcid":false,"given":"Kalyan C.","family":"Nagulapalli Venkata","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, University of Health Sciences and Pharmacy, St. Louis, MO 63010, USA"},{"name":"Department of Pharmaceutical and Administrative Sciences, Saint Louis College of Pharmacy, University of Health Sciences and Pharmacy, St. Louis, MO 63010, USA"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"103981","DOI":"10.1016\/j.langlo.2026.103981","article-title":"WHO estimates of the global, regional, and national burden of eight foodborne non-diarrhoeal enteric disease hazards, 2000\u20132021: An updated data synthesis","volume":"14","author":"Majowicz","year":"2026","journal-title":"Lancet Glob. Health"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"828","DOI":"10.1111\/j.1872-034X.2012.00988.x","article-title":"Hepatitis A outbreak associated with a revolving sushi bar in Chiba, Japan: Application of molecular epidemiology","volume":"42","author":"Tominaga","year":"2012","journal-title":"Hepatol. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1007\/s40121-025-01282-4","article-title":"Hepatitis A to E in People with HIV: A Virus-Specific Review of Prevention and Treatment","volume":"15","author":"Lin","year":"2026","journal-title":"Infect. Dis. Ther."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Honda, M., Asakura, H., Kanda, T., Somura, Y., Ishii, T., Yamana, Y., Kaneko, T., Mizutani, T., Takahashi, H., and Kumagawa, M. (2021). Male-Dominant Hepatitis A Outbreak Observed among Non-HIV-Infected Persons in the Northern Part of Tokyo, Japan. Viruses, 13.","DOI":"10.3390\/v13020207"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"taae038","DOI":"10.1093\/jtm\/taae038","article-title":"A systematic review and meta-analysis of sexually transmitted infections and blood-borne viruses in travellers","volume":"31","author":"Shiferaw","year":"2024","journal-title":"J. Travel. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1111\/hepr.13983","article-title":"Recent advances in hepatitis A virus research and clinical practice guidelines for hepatitis A virus infection in Japan","volume":"54","author":"Kanda","year":"2024","journal-title":"Hepatol. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e230","DOI":"10.3346\/jkms.2019.34.e230","article-title":"Perspectives on Acute Hepatitis A Control in Korea","volume":"34","author":"Kang","year":"2019","journal-title":"J. Korean Med. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Sasaki-Tanaka, R., Nagulapalli Venkata, K.C., Okamoto, H., Moriyama, M., and Kanda, T. (2022). Evaluation of Potential Anti-Hepatitis A Virus 3C Protease Inhibitors Using Molecular Docking. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23116044"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.35712\/aig.v2.i1.1","article-title":"Artificial intelligence and machine learning could support drug development for hepatitis A virus internal ribosomal entry sites","volume":"2","author":"Kanda","year":"2021","journal-title":"Artif. Intell. Gastroenterol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1166","DOI":"10.1021\/acs.jcim.2c01253","article-title":"ZINC-22A Free Multi-Billion-Scale Database of Tangible Compounds for Ligand Discovery","volume":"63","author":"Tingle","year":"2023","journal-title":"J. Chem. Inf. Model."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3891","DOI":"10.1021\/acs.jcim.1c00203","article-title":"AutoDock Vina 1.2.0: New Docking Methods, Expanded Force Field, and Python Bindings","volume":"61","author":"Eberhardt","year":"2021","journal-title":"J. Chem. Inf. Model."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1016\/j.jmb.2005.09.074","article-title":"Dual modes of modification of hepatitis A virus 3C protease by a serine-derived beta-lactone: Selective crystallization and formation of a functional catalytic triad in the active site","volume":"354","author":"Yin","year":"2005","journal-title":"J. Mol. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1038\/nrd4163","article-title":"The role of ligand efficiency metrics in drug discovery","volume":"13","author":"Hopkins","year":"2014","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1126\/science.285.5424.110","article-title":"Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line","volume":"285","author":"Lohmann","year":"1999","journal-title":"Science"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Kanda, T., Sasaki-Tanaka, R., Abe, H., Yokoo, T., Sakamaki, A., Hayashi, K., Kamimura, H., Tsuchiya, A., Masuzaki, R., and Kogure, H. (2025). Zinc Acetate Inhibits Hepatitis A Virus Replication: Possible Treatment for Patients with Type A Acute-on-Chronic Liver Failure. Pathogens, 14.","DOI":"10.3390\/pathogens14090882"},{"key":"ref_16","first-page":"A3.B.1","article-title":"Trypan Blue Exclusion Test of Cell Viability","volume":"111","author":"Strober","year":"2015","journal-title":"Curr. Protoc. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Sasaki-Tanaka, R., Shibata, T., Okamoto, H., Moriyama, M., and Kanda, T. (2022). Favipiravir Inhibits Hepatitis A Virus Infection in Human Hepatocytes. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23052631"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1038\/369072a0","article-title":"Picornaviral 3C cysteine proteinases have a fold similar to chymotrypsin-like serine proteinases","volume":"369","author":"Allaire","year":"1994","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2436","DOI":"10.1128\/jvi.71.3.2436-2448.1997","article-title":"The refined crystal structure of the 3C gene product from hepatitis A virus: Specific proteinase activity and RNA recognition","volume":"71","author":"Bergmann","year":"1997","journal-title":"J. Virol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1006\/viro.1999.9968","article-title":"Crystal structure of an inhibitor complex of the 3C proteinase from hepatitis A virus (HAV) and implications for the polyprotein processing in HAV","volume":"265","author":"Bergmann","year":"1999","journal-title":"Virology"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1128\/jvi.61.1.50-59.1987","article-title":"Complete nucleotide sequence of wild-type hepatitis A virus: Comparison with different strains of hepatitis A virus and other picornaviruses","volume":"61","author":"Cohen","year":"1987","journal-title":"J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2056","DOI":"10.1128\/jvi.65.4.2056-2065.1991","article-title":"Antigenic and genetic variation in cytopathic hepatitis A virus variants arising during persistent infection: Evidence for genetic recombination","volume":"65","author":"Lemon","year":"1991","journal-title":"J. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1136\/sextrans-2022-055634","article-title":"Hepatitis A and B vaccine uptake and immunisation among men who have sex with men seeking PrEP: A substudy of the ANRS IPERGAY trial","volume":"99","author":"Charreau","year":"2023","journal-title":"Sex. Transm. Infect."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1216","DOI":"10.15585\/mmwr.mm6743a5","article-title":"Update: Recommendations of the Advisory Committee on Immunization Practices for Use of Hepatitis A Vaccine for Postexposure Prophylaxis and for Preexposure Prophylaxis for International Travel","volume":"67","author":"Nelson","year":"2018","journal-title":"MMWR Morb. Mortal. Wkly. Rep."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Al Khzem, A.H., Shoaib, T.H., Mukhtar, R.M., Alturki, M.S., Gomaa, M.S., Hussein, D., Mostafa, A., Alrumaihi, L.A., Alansari, F.A., and Laabei, M. (2025). In Silico Lead Identification of Staphylococcus aureus LtaS Inhibitors: A High-Throughput Computational Pipeline Towards Prototype Development. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms262412038"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"5761","DOI":"10.1016\/j.bmc.2008.03.059","article-title":"Heteroaromatic ester inhibitors of hepatitis A virus 3C proteinase: Evaluation of mode of action","volume":"16","author":"Huitema","year":"2008","journal-title":"Bioorganic Med. Chem."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/9\/1987\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T13:51:50Z","timestamp":1788875510000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/9\/1987"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,8]]},"references-count":26,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["microorganisms14091987"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14091987","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,8]]}}}