{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:16:17Z","timestamp":1785417377063,"version":"3.56.0"},"reference-count":43,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T00:00:00Z","timestamp":1775692800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32473092"],"award-info":[{"award-number":["32473092"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"award":["32473092"],"award-info":[{"award-number":["32473092"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Science and Technology Projects of Guangzhou City","award":["2023B03J1311"],"award-info":[{"award-number":["2023B03J1311"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Coronavirus infections pose a significant threat to both human and animal health, causing widespread morbidity, mortality, and substantial economic losses. While vaccines are crucial for prevention, their efficacy is often limited by the high mutation rate of these viruses. This underscores the urgent need for anti-coronavirus drugs, particularly broad-spectrum antiviral agents. In this study, we demonstrated for the first time that Biochanin A (BCA), a bioactive isoflavonoid found in legumes, exhibits broad-spectrum antiviral activity against coronaviruses. BCA potently inhibits porcine epidemic diarrhea virus (PEDV), as well as human coronaviruses HCoV-OC43 and HCoV-229E in vitro, with EC50 values of 6.90, 2.80 and 15.4 \u03bcM, respectively. In a lethal mouse model of HCoV-OC43-induced encephalitis, oral administration of BCA (40\u201360 mg\/kg) significantly improved animal survival and reduced cerebral viral loads. Mechanistic studies revealed that BCA upregulates the AMPK\/Nrf2 signaling pathway, thereby increasing expression of the glutamate-cysteine ligase catalytic subunit (GCLC) and enhancing glutathione (GSH) biosynthesis. Our findings identify BCA as a promising host-directed antiviral agent and highlight its therapeutic potential against coronavirus infections.<\/jats:p>","DOI":"10.3390\/microorganisms14040851","type":"journal-article","created":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T09:08:12Z","timestamp":1775812092000},"page":"851","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Biochanin A Exerts Broad-Spectrum Antiviral Activity Against Coronaviruses via Activating the AMPK\/Nrf2\/GSH Pathway"],"prefix":"10.3390","volume":"14","author":[{"given":"Qisheng","family":"Lin","sequence":"first","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fan","family":"Ji","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haiyan","family":"Shen","sequence":"additional","affiliation":[{"name":"Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiajing","family":"He","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donglan","family":"Liu","sequence":"additional","affiliation":[{"name":"Guangzhou National Laboratory, Guangzhou International Bio-Island, Guangzhou 510320, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fang","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510182, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyu","family":"Cheng","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5221-9771","authenticated-orcid":false,"given":"Weisan","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC 3086, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fengxiang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education of China), School of Chemistry and Pharmaceutical Science, Guangxi Normal University, Guilin 541004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zifeng","family":"Yang","sequence":"additional","affiliation":[{"name":"Guangzhou National Laboratory, Guangzhou International Bio-Island, Guangzhou 510320, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2978-1248","authenticated-orcid":false,"given":"Jianxin","family":"Chen","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1016\/S0140-6736(20)30183-5","article-title":"Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China","volume":"395","author":"Huang","year":"2020","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1038\/nrmicro.2016.81","article-title":"SARS and MERS: Recent insights into emerging coronaviruses","volume":"14","author":"Falzarano","year":"2016","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1350","DOI":"10.3201\/eid1803.120002","article-title":"New variants of porcine epidemic diarrhea virus, China","volume":"18","author":"Li","year":"2012","journal-title":"Emerg. Infect. Dis."},{"key":"ref_4","first-page":"994","article-title":"Remdesivir for the Treatment of Covid-19 \u2212 Preliminary Report","volume":"383","author":"Beigel","year":"2020","journal-title":"Reply. N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"n2422","DOI":"10.1136\/bmj.n2422","article-title":"Covid-19: Molnupiravir reduces risk of hospital admission or death by 50% in patients at risk, MSD reports","volume":"375","author":"Mahase","year":"2021","journal-title":"BMJ"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"n2713","DOI":"10.1136\/bmj.n2713","article-title":"Covid-19: Pfizer\u2019s paxlovid is 89% effective in patients at risk of serious illness, company reports","volume":"375","author":"Mahase","year":"2021","journal-title":"BMJ"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Huan, C., Xu, W., Ni, B., Guo, T., Pan, H., Jiang, L., Li, L., Yao, J., and Gao, S. (2021). Epigallocatechin-3-Gallate, the Main Polyphenol in Green Tea, Inhibits Porcine Epidemic Diarrhea Virus In Vitro. Front. Pharmacol., 12.","DOI":"10.3389\/fphar.2021.628526"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1467","DOI":"10.1007\/s00705-017-3259-7","article-title":"Glycyrrhizin inhibits porcine epidemic diarrhea virus infection and attenuates the proinflammatory responses by inhibition of high mobility group box-1 protein","volume":"162","author":"Huan","year":"2017","journal-title":"Arch. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Li, Z., Cao, H., Cheng, Y., Zhang, X., Zeng, W., Sun, Y., Chen, S., He, Q., and Han, H. (2020). Inhibition of Porcine Epidemic Diarrhea Virus Replication and Viral 3C-Like Protease by Quercetin. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21218095"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Kwon, E.B., Kim, B., Kim, Y.S., and Choi, J.G. (2024). Anastrozole Protects against Human Coronavirus Infection by Ameliorating the Reactive Oxygen Species-Mediated Inflammatory Response. Antioxidants, 13.","DOI":"10.3390\/antiox13010116"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1186\/s13567-023-01139-z","article-title":"Porcine epidemic diarrhea virus activates PERK-ROS axis to benefit its replication in Vero E6 cells","volume":"54","author":"Zhou","year":"2023","journal-title":"Vet. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.ceb.2017.01.005","article-title":"AMPK signalling in health and disease","volume":"45","author":"Carling","year":"2017","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.freeradbiomed.2022.07.014","article-title":"AMPK and NRF2: Interactive players in the same team for cellular homeostasis?","volume":"190","author":"Petsouki","year":"2022","journal-title":"Free Radic. Biol. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.mam.2008.08.009","article-title":"Structure, function, and post-translational regulation of the catalytic and modifier subunits of glutamate cysteine ligase","volume":"30","author":"Franklin","year":"2009","journal-title":"Mol. Asp. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2291","DOI":"10.1083\/jcb.201804161","article-title":"Glutathione metabolism in cancer progression and treatment resistance","volume":"217","author":"Bansal","year":"2018","journal-title":"J. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1093\/oxfordjournals.aje.a118408","article-title":"A simple method of estimating fifty per cent endpoints","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J. Epidemiol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Su, L., Zhang, Z., Zhang, S., He, J., Chen, Y., Wu, G., Zhang, Z., Chen, W., Zhang, M., and Chen, J. (2025). Early boosting of p38 MAPK signaling pathway by lycorine hydrochloride potently inhibits PRRSV proliferation in primary and established cells. Front. Microb., 16.","DOI":"10.3389\/fmicb.2025.1664973"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"105635","DOI":"10.1016\/j.antiviral.2023.105635","article-title":"Artesunate inhibits PDE4 leading to intracellular cAMP accumulation, reduced ERK\/MAPK signaling, and blockade of influenza A virus vRNP nuclear export","volume":"215","author":"Yang","year":"2023","journal-title":"Antivir. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e2415030","DOI":"10.1002\/adma.202415030","article-title":"Self-Sustained Biophotocatalytic Nano-Organelle Reactors with Programmable DNA Switches for Combating Tumor Metastasis","volume":"37","author":"Han","year":"2025","journal-title":"Adv. Mater."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e0148721","DOI":"10.1128\/JVI.01487-21","article-title":"The Antimalaria Drug Artesunate Inhibits Porcine Reproductive and Respiratory Syndrome Virus Replication by Activating AMPK and Nrf2\/HO-1 Signaling Pathways","volume":"96","author":"Long","year":"2022","journal-title":"J. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Lin, Y., Zang, R., Ma, Y., Wang, Z., Li, L., Ding, S., Zhang, R., Wei, Z., Yang, J., and Wang, X. (2021). Xanthohumol Is a Potent Pan-Inhibitor of Coronaviruses Targeting Main Protease. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222212134"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1038\/s42003-024-06418-9","article-title":"Dorsomorphin inhibits AMPK, upregulates Wnt and Foxo genes and promotes the activation of dormant follicles","volume":"7","author":"Madsen","year":"2024","journal-title":"Commun. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2431","DOI":"10.1056\/NEJMra032498","article-title":"The severe acute respiratory syndrome","volume":"349","author":"Peiris","year":"2003","journal-title":"N. Engl. J. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"198045","DOI":"10.1016\/j.virusres.2020.198045","article-title":"Porcine epidemic diarrhea virus (PEDV): An update on etiology, transmission, pathogenesis, and prevention and control","volume":"286","author":"Jung","year":"2020","journal-title":"Virus Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.biocel.2006.07.001","article-title":"Free radicals and antioxidants in normal physiological functions and human disease","volume":"39","author":"Valko","year":"2007","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1291","DOI":"10.1152\/physrev.00034.2023","article-title":"NADPH Oxidases: Redox Regulation of Cell Homeostasis & Disease","volume":"105","author":"Lopes","year":"2025","journal-title":"Physiol. Rev."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1016\/j.freeradbiomed.2021.06.018","article-title":"Upregulation of oxidative stress gene markers during SARS-CoV-2 viral infection","volume":"172","author":"Mdkhana","year":"2021","journal-title":"Free Radic. Biol. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"8159562","DOI":"10.1155\/2019\/8159562","article-title":"Oxidative Stress in Microbial Diseases: Pathogen, Host, and Therapeutics","volume":"2019","author":"Novaes","year":"2019","journal-title":"Oxid. Med. Cell Longev."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1016\/j.bcp.2012.11.016","article-title":"The Nrf2 cell defence pathway: Keap1-dependent and -independent mechanisms of regulation","volume":"85","author":"Bryan","year":"2013","journal-title":"Biochem. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1016\/j.neuron.2024.12.016","article-title":"CRISPRi-based screens in iAssembloids to elucidate neuron-glia interactions","volume":"113","author":"Li","year":"2025","journal-title":"Neuron"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"618","DOI":"10.1016\/j.molcel.2013.08.003","article-title":"Phosphorylation of p62 Activates the Keap1-Nrf2 Pathway during Selective Autophagy","volume":"51","author":"Ichimura","year":"2013","journal-title":"Mol. Cell"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1038\/ncb2021","article-title":"The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1","volume":"12","author":"Komatsu","year":"2010","journal-title":"Nat. Cell Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1186\/1475-4924-2-28","article-title":"Complexes between the LKB1 tumor suppressor, STRAD alpha\/beta and MO25 alpha\/beta are upstream kinases in the AMP-activated protein kinase cascade","volume":"2","author":"Hawley","year":"2003","journal-title":"J. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.cmet.2005.06.005","article-title":"Ca2+\/calmodulin-dependent protein kinase kinase-beta acts upstream of AMP-activated protein kinase in mammalian cells","volume":"2","author":"Woods","year":"2005","journal-title":"Cell Metab."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1038\/nrm.2017.95","article-title":"AMPK: Guardian of metabolism and mitochondrial homeostasis","volume":"19","author":"Herzig","year":"2018","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"9943","DOI":"10.1073\/pnas.87.24.9943","article-title":"Intracellular thiols regulate activation of nuclear factor kappa B and transcription of human immunodeficiency virus","volume":"87","author":"Staal","year":"1990","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"928","DOI":"10.1016\/S0891-5849(03)00023-6","article-title":"Inhibition of influenza infection by glutathione","volume":"34","author":"Cai","year":"2003","journal-title":"Free Radic. Biol. Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.mam.2008.09.001","article-title":"GSH and analogs in antiviral therapy","volume":"30","author":"Fraternale","year":"2009","journal-title":"Mol. Asp. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1016\/j.jare.2025.04.021","article-title":"Design, synthesis and activity evaluation of dual-target inhibitors against papain-like and main proteases of porcine epidemic diarrhea virus","volume":"80","author":"Chen","year":"2025","journal-title":"J. Adv. Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"156699","DOI":"10.1016\/j.phymed.2025.156699","article-title":"Biochanin A mitigates colitis by inhibiting ferroptosis-mediated intestinal barrier dysfunction, oxidative stress, and inflammation via the JAK2\/STAT3 signaling pathway","volume":"141","author":"Deng","year":"2025","journal-title":"Phytomedicine"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e41980","DOI":"10.1016\/j.heliyon.2025.e41980","article-title":"SARS-CoV-2 drug resistance and therapeutic approaches","volume":"11","author":"Batool","year":"2025","journal-title":"Heliyon"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"104559","DOI":"10.1016\/j.ebiom.2023.104559","article-title":"Global prevalence of SARS-CoV-2 3CL protease mutations associated with nirmatrelvir or ensitrelvir resistance","volume":"91","author":"Ip","year":"2023","journal-title":"EBioMedicine"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Szemiel, A.M., Merits, A., Orton, R.J., MacLean, O.A., Pinto, R.M., Wickenhagen, A., Lieber, G., Turnbull, M.L., Wang, S., and Furnon, W. (2021). In vitro selection of Remdesivir resistance suggests evolutionary predictability of SARS-CoV-2. PLoS Pathog., 17.","DOI":"10.1371\/journal.ppat.1009929"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/4\/851\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T04:28:19Z","timestamp":1775881699000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/4\/851"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,9]]},"references-count":43,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["microorganisms14040851"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14040851","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,9]]}}}