{"doi":"10.3390/pharmaceutics17121524","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","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>","journal":"Pharmaceutics","year":2025,"id":648607,"datarank":0.17681582500380735,"base_score":1.0986122886681096,"endowment":1.0986122886681096,"self_citation_contribution":0.16479184330021646,"citation_network_contribution":0.012023981703590886,"self_endowment_contribution":0.16479184330021646,"citer_contribution":0.012023981703590886,"corpus_percentile":null,"corpus_rank":null,"citation_count":2,"citer_count":2,"citers_with_citation_signal":1,"citers_with_endowment":1,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":401561,"name":"Yibo Chen","orcid":"0000-0002-0178-9715","position":1,"is_corresponding":false},{"id":1690432,"name":"Binghui Xia","orcid":"0000-0002-8771-2881","position":2,"is_corresponding":false},{"id":1690435,"name":"Zimeng Cheng","orcid":"0009-0001-5401-9319","position":3,"is_corresponding":false},{"id":63825,"name":"Ping Zhao","orcid":"0000-0002-5618-0472","position":4,"is_corresponding":false},{"id":1581645,"name":"Zhongtian Qi","orcid":null,"position":5,"is_corresponding":false},{"id":1690440,"name":"Yongzhe Zhu","orcid":"0000-0002-1529-6807","position":6,"is_corresponding":false},{"id":757473,"name":"Zhiwei He","orcid":"0000-0001-7264-2019","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"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","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>","is_dataset_classified":null,"base_score":1.0986122886681096,"endowment":1.0986122886681096,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"41471039","pmcid":"PMC12736548","openalex_id":"https://openalex.org/W4416736354","authors":[],"funders":[{"funder_name":"National Natural Science Foundation of China","grant_id":"31770181","title":null}],"total_grants":1,"fwci":0.8477,"citation_percentile":0.75042423,"influential_citations":0,"citation_trend":[{"year":2026,"count":2}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://www.mdpi.com/1999-4923/17/12/1524/pdf?version=1764233884","host_type":"journal"},{"url":"https://www.mdpi.com/1999-4923/17/12/1524/pdf?version=1764233884","host_type":"publisher"},{"url":"https://www.mdpi.com/1999-4923/17/12/1524/pdf","host_type":"publisher"},{"url":"https://doi.org/10.3390/pharmaceutics17121524","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/41471039","host_type":"repository"},{"url":"https://doaj.org/article/2a0e49e76fff4677b0ce8f6755d3d1bf","host_type":"repository"},{"url":"https://pmc.ncbi.nlm.nih.gov/articles/PMC12736548/","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC12736548","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC12736548?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["Protein Degradation and Inhibitors","Mosquito-borne diseases and control","Endoplasmic Reticulum Stress and Disease"],"mesh_terms":[],"keywords":["Gene knockdown","Virus","Ubiquitin ligase","In vivo","Small interfering RNA","In vitro","Protein degradation","RNA interference","Sindbis virus","Japanese encephalitis virus (JEV)","Non-structural Proteins","Broad-spectrum Antiviral Activity","Cereblon E3 Ligase Modulators (Celmod)","G1-To-S Phase Transition 1 (Gspt1)"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"gen"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-10T02:47:17.305572Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}