{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T05:01:45Z","timestamp":1787029305268,"version":"build-2736575974"},"reference-count":33,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2025,11,27]],"date-time":"2025-11-27T00:00:00Z","timestamp":1764201600000},"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":["31770181"],"award-info":[{"award-number":["31770181"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Background: Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, remains a leading cause of viral encephalitis. Current management is largely supportive, with no specific antivirals. This study evaluated the antiviral efficacy and mechanism of action of CC-90009 against JEV in vitro and in vivo. Methods: Five targeted protein degraders (TPDs) were screened for anti-JEV activity in the human neuroblastoma cell line SH-SY5Y. Time-of-addition, binding, and endocytosis assays were used to delineate the phase of action of CC-90009, a cereblon (CRBN) E3 ligase modulator (CELMoD) and molecular glue degrader. Small interfering RNA knockdown and co-immunoprecipitation (Co-IP) confirmed targets essential for its antiviral effects. The broad-spectrum activity of CC-90009 against other mosquito-borne viruses was also evaluated. In vivo efficacy was tested in a murine JEV model. Results: Of the five TPDs tested, only CC-90009 significantly inhibited JEV infection in SH-SY5Y cells, acting during both viral entry and post-entry phases without reducing adsorbed or internalised virions. CC-90009 reduced JEV RNA and non-structural protein accumulation. Knockdown of G1-to-S phase transition 1 (GSPT1), a key target of CC-90009, suppressed JEV infection and translation; Co-IP confirmed GSPT1 interaction with JEV non-structural protein 5 (NS5). CC-90009 disrupted JEV translation and replication by inducing proteasomal degradation of the GSPT1\/NS5 complex, further demonstrating its broad-spectrum antiviral activity through the effective inhibition of West Nile virus and chikungunya virus. In vivo, it protected mice from JEV-induced mortality, reducing viral load, antigen levels, and brain pathology. Conclusions: CC-90009 exerts potent anti-JEV activity both in vitro and in vivo by inducing proteasomal degradation of the GSPT1\/NS5 complex, thereby disrupting viral translation and replication. This targeted protein degradation strategy represents a novel host-directed antiviral approach with promising therapeutic potential against mosquito-borne viral encephalitis.<\/jats:p>","DOI":"10.3390\/pharmaceutics17121524","type":"journal-article","created":{"date-parts":[[2025,11,27]],"date-time":"2025-11-27T10:25:16Z","timestamp":1764239116000},"page":"1524","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["CC-90009, a Cereblon E3 Ligase Modulator, Exhibits Antiviral Efficacy Against JEV In Vitro and In Vivo via Targeted Degradation of GSPT1 and Viral NS5 Protein"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-8970-5513","authenticated-orcid":false,"given":"Zhiwei","family":"He","sequence":"first","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-5705-8973","authenticated-orcid":false,"given":"Yibo","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8771-2881","authenticated-orcid":false,"given":"Binghui","family":"Xia","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-5401-9319","authenticated-orcid":false,"given":"Zimeng","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Shanghai Key Laboratory of Medical Bioprotection, Second Military Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biological Defense, Ministry of Education, Second Military Medical University, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhongtian","family":"Qi","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Shanghai Key Laboratory of Medical Bioprotection, Second Military Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biological Defense, Ministry of Education, Second Military Medical University, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1529-6807","authenticated-orcid":false,"given":"Yongzhe","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Shanghai Key Laboratory of Medical Bioprotection, Second Military Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biological Defense, Ministry of Education, Second Military Medical University, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Zhu, Y., Chen, S., Lurong, Q., and Qi, Z. (2023). Recent Advances in Antivirals for Japanese Encephalitis Virus. Viruses, 15.","DOI":"10.3390\/v15051033"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Guo, J., Mi, Y., Guo, Y., Bai, Y., Wang, M., Wang, W., and Wang, Y. (2024). Current Advances in Japanese Encephalitis Virus Drug Development. Viruses, 16.","DOI":"10.3390\/v16020202"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1186\/s13071-025-06843-7","article-title":"Mosquito vector competence for Japanese encephalitis virus: A systematic review and meta-analysis update","volume":"18","author":"Edache","year":"2025","journal-title":"Parasites Vectors"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Srivastava, K.S., Jeswani, V., Pal, N., Bohra, B., Vishwakarma, V., Bapat, A.A., Patnaik, Y.P., Khanna, N., and Shukla, R. (2023). Japanese Encephalitis Virus: An Update on the Potential Antivirals and Vaccines. Vaccines, 11.","DOI":"10.3390\/vaccines11040742"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"199120","DOI":"10.1016\/j.virusres.2023.199120","article-title":"Virus-host Interactions in Early Japanese Encephalitis Virus Infection","volume":"331","author":"Zhu","year":"2023","journal-title":"Virus Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"106268","DOI":"10.1016\/j.antiviral.2025.106268","article-title":"Comprehensive insights into Japanese encephalitis virus: From molecular characterization to advanced detection and vaccine strategies","volume":"243","author":"Chugh","year":"2025","journal-title":"Antivir. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1227","DOI":"10.2217\/fmb-2016-0002","article-title":"Caveolin-1-mediated Japanese encephalitis virus entry requires a two-step regulation of actin reorganization","volume":"11","author":"Xu","year":"2016","journal-title":"Future Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1995","DOI":"10.1099\/jgv.0.001995","article-title":"Japanese encephalitis virus hijacks ER-associated degradation regulators for its replication","volume":"105","author":"Sarkar","year":"2024","journal-title":"J. Gen. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1739","DOI":"10.1007\/s00705-022-05481-z","article-title":"Molecular pathogenesis of Japanese encephalitis and possible therapeutic strategies","volume":"167","author":"Kumar","year":"2022","journal-title":"Arch. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Lu, G., and Gong, P. (2013). Crystal Structure of the full-length Japanese encephalitis virus NS5 reveals a conserved methyltransferase-polymerase interface. PLoS Pathog., 9.","DOI":"10.2210\/pdb4k6m\/pdb"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1038\/s41568-025-00817-8","article-title":"Targeted protein degradation for cancer therapy","volume":"25","author":"Hinterndorfer","year":"2025","journal-title":"Nat. Rev. Cancer"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6648","DOI":"10.1021\/acs.jmedchem.9b01928","article-title":"Discovery of CRBN E3 Ligase Modulator CC-92480 for the Treatment of Relapsed and Refractory Multiple Myeloma","volume":"63","author":"Hansen","year":"2020","journal-title":"J. Med. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1016\/j.chembiol.2023.02.005","article-title":"Discovery and characterization of a selective IKZF2 glue degrader for cancer immunotherapy","volume":"30","author":"Bonazzi","year":"2023","journal-title":"Cell Chem. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1182\/blood.2020008676","article-title":"CC-90009, a novel cereblon E3 ligase modulator, targets acute myeloid leukemia blasts and leukemia stem cells","volume":"137","author":"Surka","year":"2021","journal-title":"Blood"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1158\/1535-7163.MCT-23-0655","article-title":"Preclinical Evaluation of Bavdegalutamide (ARV-110), a Novel PROteolysis TArgeting Chimera Androgen Receptor Degrader","volume":"24","author":"Snyder","year":"2025","journal-title":"Mol. Cancer Ther."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3549","DOI":"10.1158\/1078-0432.CCR-23-3465","article-title":"Oral Estrogen Receptor PROTAC Vepdegestrant (ARV-471) Is Highly Efficacious as Monotherapy and in Combination with CDK4\/6 or PI3K\/mTOR Pathway Inhibitors in Preclinical ER+ Breast Cancer Models","volume":"30","author":"Gough","year":"2024","journal-title":"Clin. Cancer Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1038\/s41392-022-00966-4","article-title":"Targeted protein degradation: Mechanisms, strategies and application","volume":"7","author":"Zhao","year":"2022","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1835","DOI":"10.1021\/acs.jmedchem.0c01489","article-title":"CC-90009: A Cereblon E3 Ligase Modulating Drug That Promotes Selective Degradation of GSPT1 for the Treatment of Acute Myeloid Leukemia","volume":"64","author":"Hansen","year":"2021","journal-title":"J. Med. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e2201208119","DOI":"10.1073\/pnas.2201208119","article-title":"Proximity interactome analysis of Lassa polymerase reveals eRF3a\/GSPT1 as a druggable target for host-directed antivirals","volume":"119","author":"Fang","year":"2022","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"110544","DOI":"10.1016\/j.celrep.2022.110544","article-title":"Functional interactomes of the Ebola virus polymerase identified by proximity proteomics in the context of viral replication","volume":"38","author":"Fang","year":"2022","journal-title":"Cell Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e70369","DOI":"10.1002\/jmv.70369","article-title":"Clathrin-Independent Carriers\/Glycosylphosphatidylinositol-Anchored-Protein-Enriched Endosomal Compartment Endocytic Pathway Is Critical for Enterovirus A71 Entry Into Human Oral Epidermoid Carcinoma KB Cells","volume":"97","author":"He","year":"2025","journal-title":"J. Med. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e28915","DOI":"10.1002\/jmv.28915","article-title":"Enterovirus 71 enters human brain microvascular endothelial cells through an ARF6-mediated endocytic pathway","volume":"95","author":"Zhu","year":"2023","journal-title":"J. Med. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1330","DOI":"10.1080\/22221751.2020.1757388","article-title":"Ezrin is essential for the entry of Japanese encephalitis virus into the human brain microvascular endothelial cells","volume":"9","author":"Liu","year":"2020","journal-title":"Emerg. Microbes Infect."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Hellen, C.U.T. (2018). Translation Termination and Ribosome Recycling in Eukaryotes. Cold Spring Harb. Perspect. Biol., 10.","DOI":"10.1101\/cshperspect.a032656"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Goh, J.Z.H., De Hayr, L., Khromykh, A.A., and Slonchak, A. (2024). The Flavivirus Non-Structural Protein 5 (NS5): Structure, Functions, and Targeting for Development of Vaccines and Therapeutics. Vaccines, 12.","DOI":"10.3390\/vaccines12080865"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1002\/jcb.30352","article-title":"Drug repurposing toward the inhibition of RNA-dependent RNA polymerase of various flaviviruses through computational study","volume":"124","author":"Murali","year":"2023","journal-title":"J. Cell. Biochem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.antiviral.2008.06.007","article-title":"The flavivirus polymerase as a target for drug discovery","volume":"80","author":"Malet","year":"2008","journal-title":"Antivir. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1000","DOI":"10.1093\/nar\/gkab1302","article-title":"Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold","volume":"50","author":"Tan","year":"2022","journal-title":"Nucleic Acids Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1016\/j.tig.2024.04.003","article-title":"Positive-strand RNA virus genome replication organelles: Structure, assembly, control","volume":"40","author":"Nishikiori","year":"2024","journal-title":"Trends Genet."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"860","DOI":"10.1038\/nrmicro2655","article-title":"Viral subversion of the host protein synthesis machinery","volume":"9","author":"Walsh","year":"2011","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Liu, C., Peng, H., Chen, P., Li, Y., Deng, Z., Li, S., Yang, T., Liu, K., Wang, Z., and Liu, L. (2025). GSPT1 degraders: Research progress, development strategies and challenges. Bioorg. Med. Chem., 131.","DOI":"10.1016\/j.bmc.2025.118390"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"13169","DOI":"10.1021\/acs.jmedchem.5c00783","article-title":"Research Progress in Targeting GSPT1: Molecular Glues, Bifunctional Degraders, and Antibody-Enabled Molecular Glues for Cancer Therapy","volume":"68","author":"Cheng","year":"2025","journal-title":"J. Med. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1154","DOI":"10.1016\/j.chom.2023.05.030","article-title":"Generation of host-directed and virus-specific antivirals using targeted protein degradation promoted by small molecules and viral RNA mimics","volume":"31","author":"Zhao","year":"2023","journal-title":"Cell Host Microbe"}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/17\/12\/1524\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,27]],"date-time":"2025-11-27T10:33:32Z","timestamp":1764239612000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/17\/12\/1524"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,27]]},"references-count":33,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["pharmaceutics17121524"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics17121524","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,27]]}}}