{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T18:26:25Z","timestamp":1787163985444,"version":"build-2736575974"},"reference-count":38,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T00:00:00Z","timestamp":1771286400000},"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":"publisher","award":["82574419"],"award-info":[{"award-number":["82574419"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82130101"],"award-info":[{"award-number":["82130101"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82404430"],"award-info":[{"award-number":["82404430"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Major scientific and technological projects of Guangdong Province","award":["2023B1111050008)"],"award-info":[{"award-number":["2023B1111050008)"]}]},{"name":"Post Scientist Fund awarded by Chinese Academy of Traditional Chinese Medicine","award":["ZZ13-035-02"],"award-info":[{"award-number":["ZZ13-035-02"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Influenza virus infection remains a major global health burden, with severe disease outcomes driven not only by viral replication but also by excessive host inflammatory responses. Current antiviral therapies predominantly target viral components and fail to adequately control virus-induced hyperinflammation. In this study, we report a dual-target therapeutic strategy integrating direct antiviral activity with host-directed immunomodulation. Using a molecular hybridization approach, we designed and synthesized several dual-target inhibitors simultaneously targeting the influenza virus PB2 cap-binding subunit and host JAK2 kinase. Among them, PB05 emerged as the most promising candidate and was systematically evaluated in vitro and in vivo. PB05 exhibited potent broad-spectrum antiviral activity against influenza A viruses, with nanomolar EC50 values. Mechanistic studies demonstrated that PB05 directly binds to the PB2 cap-binding domain, thereby disrupting viral cap-snatching and RNA synthesis. In parallel, PB05 inhibited JAK\u2013STAT signaling by suppressing STAT2 phosphorylation and downstream ISRE-mediated transcription, leading to a marked reduction in pro-inflammatory cytokine production, including IL-6, IL-1\u03b2, and IFN-\u03b2, in infected or stimulated immune cells. In a lethal influenza A\/PR\/8\/34 (H1N1) mouse model, oral administration of PB05 at 100 mg\/kg (twice daily) markedly decreased lung viral titers, attenuated pulmonary tissue damage and edema, and moderated excessive inflammatory responses. Collectively, these findings identify PB05 as a dual PB2\/JAK2 inhibitor that effectively couples antiviral efficacy with immunomodulatory activity, promoting a therapeutic strategy for the treatment of severe influenza and other viral diseases associated with excessive inflammation.<\/jats:p>","DOI":"10.3390\/molecules31040696","type":"journal-article","created":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T13:15:35Z","timestamp":1771334135000},"page":"696","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Dual Inhibition of PB2 and JAK2 for Influenza: A Strategy Combining Antiviral and Host-Directed Immune Modulation"],"prefix":"10.3390","volume":"31","author":[{"given":"Binhao","family":"Rong","sequence":"first","affiliation":[{"name":"Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujian","family":"Yang","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Small Molecule Drug Discovery and Synthesis, Department of Chemistry, Shenzhen Grubbs Institute and Medi-X Pingshan, Southern University of Science and Technology, Shenzhen 518000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kunyu","family":"Lu","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xingyu","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Computer Science and Engineering, Southern University of Science and Technology, Shenzhen 518000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peisen","family":"Zheng","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Small Molecule Drug Discovery and Synthesis, Department of Chemistry, Shenzhen Grubbs Institute and Medi-X Pingshan, Southern University of Science and Technology, Shenzhen 518000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinxin","family":"Lin","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanmei","family":"Wen","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Small Molecule Drug Discovery and Synthesis, Department of Chemistry, Shenzhen Grubbs Institute and Medi-X Pingshan, Southern University of Science and Technology, Shenzhen 518000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shudong","family":"Lin","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Small Molecule Drug Discovery and Synthesis, Department of Chemistry, Shenzhen Grubbs Institute and Medi-X Pingshan, Southern University of Science and Technology, Shenzhen 518000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0009-5460","authenticated-orcid":false,"given":"Xinshan","family":"Deng","sequence":"additional","affiliation":[{"name":"Hunan Provincial Key Laboratory of the Research and Development of Novel Pharmaceutical Preparations, Changsha Medical University, Changsha 410219, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5738-3511","authenticated-orcid":false,"given":"Qifan","family":"Zhou","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Small Molecule Drug Discovery and Synthesis, Department of Chemistry, Shenzhen Grubbs Institute and Medi-X Pingshan, Southern University of Science and Technology, Shenzhen 518000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6346-5006","authenticated-orcid":false,"given":"Shuwen","family":"Liu","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China"},{"name":"NMPA Key Laboratory for Research and Evaluation of Drug Metabolism, Department of Pharmacy, Pingshan Hospital, Southern Medical University, Shenzhen 518100, China"},{"name":"State Key Laboratory of Multi-Organ Injury Prevention and Treatment, MOE Key Laboratory of Infectious Diseases Research in South China, MOE Innovation Center for Medical Basic Research on Inflammation and Immune Related Diseases, Southern Medical University, Guangzhou 510515, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e15352","DOI":"10.15252\/emmm.202115352","article-title":"Seasonality of influenza and other respiratory viruses","volume":"14","author":"Neumann","year":"2022","journal-title":"EMBO Mol. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1007\/s40121-021-00534-3","article-title":"A systematic review of influenza epidemiology and surveillance in the Eastern Mediterranean and North African region","volume":"11","author":"Soudani","year":"2022","journal-title":"Infect. Dis. Ther."},{"key":"ref_3","first-page":"621","article-title":"Severe influenza patients who received ventilator management during the 2024 to 2025 major influenza-endemic season in a tertiary hospital in Japan","volume":"18","author":"Seki","year":"2025","journal-title":"Int. Med. Case Rep. J."},{"key":"ref_4","first-page":"153","article-title":"Influenza A: Understanding the viral life cycle","volume":"82","author":"Samji","year":"2009","journal-title":"Yale J. Biol. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1038\/s41572-018-0002-y","article-title":"Influenza","volume":"4","author":"Krammer","year":"2018","journal-title":"Nat. Rev. Dis. Primer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"22039","DOI":"10.1021\/acs.jmedchem.4c01979","article-title":"Discovery of novel thiophene-based baloxavir derivatives as potent cap-dependent endonuclease inhibitors for influenza treatment","volume":"67","author":"Chen","year":"2024","journal-title":"J. Med. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1111\/nyas.12462","article-title":"Continuing challenges in influenza","volume":"1323","author":"Webster","year":"2014","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1007\/978-981-10-8456-0_5","article-title":"Structure and function of influenza virus ribonucleoprotein","volume":"88","author":"Lo","year":"2018","journal-title":"Subcell. Biochem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"913","DOI":"10.1056\/NEJMoa1716197","article-title":"Baloxavir marboxil for uncomplicated influenza in adults and adolescents","volume":"379","author":"Hayden","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"107512","DOI":"10.1016\/j.pharmthera.2020.107512","article-title":"Favipiravir, an anti-influenza drug against life-threatening RNA virus infections","volume":"209","author":"Shiraki","year":"2020","journal-title":"Pharmacol. Ther."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1609","DOI":"10.1093\/nar\/gkaa1294","article-title":"Structural insights into the substrate specificity of the endonuclease activity of the influenza virus cap-snatching mechanism","volume":"49","author":"Kumar","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"104545","DOI":"10.1016\/j.antiviral.2019.104545","article-title":"Antivirals targeting the polymerase complex of influenza viruses","volume":"169","author":"Mifsud","year":"2019","journal-title":"Antivir. Res."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Gregor, J., Radilov\u00e1, K., Brynda, J., Fanfrl\u00edk, J., Konvalinka, J., and Ko\u017e\u00ed\u0161ek, M. (2021). Structural and thermodynamic analysis of the resistance development to pimodivir (VX-787), the clinical inhibitor of cap binding to PB2 subunit of influenza A polymerase. Molecules, 26.","DOI":"10.3390\/molecules26041007"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1016\/S1473-3099(23)00743-0","article-title":"Safety and efficacy of onradivir in adults with acute uncomplicated influenza A infection: A multicentre, double-llind, randomised, placebo-controlled, phase 2 trial","volume":"24","author":"Yang","year":"2024","journal-title":"Lancet Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1016\/S2213-2600(25)00046-3","article-title":"Efficacy and safety of onradivir in adults with acute uncomplicated influenza A infection in China: A multicentre, double-blind, randomised, placebo-controlled and oseltamivir-controlled, phase 3 trial","volume":"13","author":"Yang","year":"2025","journal-title":"Lancet Respir. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1038\/s41392-025-02178-y","article-title":"Deep insight into cytokine storm: From pathogenesis to treatment","volume":"10","author":"Nie","year":"2025","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1186\/s13054-019-2539-x","article-title":"Influenza virus-related critical illness: Pathophysiology and epidemiology","volume":"23","author":"Kalil","year":"2019","journal-title":"Crit. Care Lond. Engl."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Chang, D., Dela Cruz, C., and Sharma, L. (2024). Beneficial and detrimental effects of cytokines during influenza and COVID-19. Viruses, 16.","DOI":"10.3390\/v16020308"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Liao, G., Xia, M., Jiang, Y., Chen, H., Liao, W., Peng, J., and Cai, S. (2023). Prospective observational study of baloxavir marboxil in adults and adolescents with uncomplicated influenza from China. Front. Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1292735"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1038\/nri3665","article-title":"Innate immunity to influenza virus infection","volume":"14","author":"Iwasaki","year":"2014","journal-title":"Nat. Rev. Immunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1038\/nature12862","article-title":"Pan-viral specificity of IFN-induced genes reveals new roles for cGAS in innate immunity","volume":"505","author":"Schoggins","year":"2014","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Malik, G., and Zhou, Y. (2020). Innate immune sensing of influenza A virus. Viruses, 12.","DOI":"10.3390\/v12070755"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1038\/s41577-024-01029-1","article-title":"Defining the balance between optimal immunity and immunopathology in influenza virus infection","volume":"24","author":"Nguyen","year":"2024","journal-title":"Nat. Rev. Immunol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Gui, R., and Chen, Q. (2021). Molecular events involved in influenza A virus-induced cell death. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.797789"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Hartshorn, K.L. (2020). Innate immunity and influenza A virus pathogenesis: Lessons for COVID-19. Front. Cell. Infect. Microbiol., 10.","DOI":"10.3389\/fcimb.2020.563850"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1159\/000508247","article-title":"JAK inhibition as a new treatment strategy for patients with COVID-19","volume":"181","author":"Seif","year":"2020","journal-title":"Int. Arch. Allergy Immunol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"An, W., Lakhina, S., Leong, J., Rawat, K., and Husain, M. (2024). Host innate antiviral response to influenza A virus infection: From viral sensing to antagonism and escape. Pathogens, 13.","DOI":"10.3390\/pathogens13070561"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1038\/s41392-023-01468-7","article-title":"Evolving cognition of the JAK-STAT signaling pathway: Autoimmune disorders and cancer","volume":"8","author":"Xue","year":"2023","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1111\/j.1600-065X.2008.00754.x","article-title":"Janus kinases in immune cell signaling","volume":"228","author":"Ghoreschi","year":"2009","journal-title":"Immunol. Rev."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Yin, Q., Wang, L., Yu, H., Chen, D., Zhu, W., and Sun, C. (2021). Pharmacological effects of polyphenol phytochemicals on the JAK-STAT signaling pathway. Front. Pharmacol., 12.","DOI":"10.3389\/fphar.2021.716672"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Wang, J., Prinz, R.A., Liu, X., and Xu, X. (2020). In vitro and In vivo antiviral activity of Gingerenone A on influenza A virus is mediated by targeting Janus kinase 2. Viruses, 12.","DOI":"10.3390\/v12101141"},{"key":"ref_32","first-page":"10132","article-title":"A77 1726, the active metabolite of the anti-rheumatoid arthritis drug leflunomide, inhibits influenza A virus replication in vitro and in vivo by inhibiting the activity of Janus kinases","volume":"34","author":"Wang","year":"2020","journal-title":"FASEB J. Off. Publ. Fed. Am. Soc. Exp. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1124\/jpet.114.221176","article-title":"VX-509 (Decernotinib) is a potent and selective Janus kinase 3 inhibitor that attenuates inflammation in animal models of autoimmune disease","volume":"353","author":"Mahajan","year":"2015","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1128\/MMBR.05015-11","article-title":"Into the eye of the cytokine storm","volume":"76","author":"Tisoncik","year":"2012","journal-title":"Microbiol. Mol. Biol. Rev. MMBR"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1099","DOI":"10.15252\/emmm.201606413","article-title":"IFN\u03bb is a potent anti-influenza therapeutic without the inflammatory side effects of IFN\u03b1 treatment","volume":"8","author":"Davidson","year":"2016","journal-title":"EMBO Mol. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"316","DOI":"10.7150\/thno.49713","article-title":"Immunopathogenesis and treatment of cytokine storm in COVID-19","volume":"11","author":"Kim","year":"2021","journal-title":"Theranostics"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1056\/NEJMoa2031994","article-title":"Baricitinib plus remdesivir for hospitalized adults with COVID-19","volume":"384","author":"Kalil","year":"2021","journal-title":"N. Engl. J. Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2386","DOI":"10.1182\/blood.2023021046","article-title":"Differential effects of itacitinib, fedratinib, and ruxolitinib in mouse models of hemophagocytic lymphohistiocytosis","volume":"143","author":"Keenan","year":"2024","journal-title":"Blood"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/4\/696\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T05:33:27Z","timestamp":1772084007000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/4\/696"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,17]]},"references-count":38,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["molecules31040696"],"URL":"https:\/\/doi.org\/10.3390\/molecules31040696","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,17]]}}}