{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T01:47:38Z","timestamp":1789523258269,"version":"build-2803163510"},"reference-count":40,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2023,9,27]],"date-time":"2023-09-27T00:00:00Z","timestamp":1695772800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Mahidol University","award":["NDFR 33\/2564"],"award-info":[{"award-number":["NDFR 33\/2564"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Every year, dengue virus (DENV) affects millions of people. Currently, there are no approved drugs for the treatment of DENV infection. Autophagy is a conserved degradation process that was shown to be induced by DENV infection and required for optimal DENV replication. The modulation of autophagy is, therefore, considered an attractive target to treat DENV infection. This study carried out a high-content image screen analysis using Crispr-Cas9 GFP-LC3 knocked-in HeLa cells of a compound library synthesized from or inspired by natural products and their biocongener precursors to discover novel autophagy inhibitors. The screen identified Ka-003 as the most effective compound for decreasing the number of autophagic vacuoles inside cells upon autophagy induction. Ka-003 could inhibit autophagy in a dose-dependent manner at low micromolar concentrations. More importantly, Ka-003 demonstrated the concentration-dependent inhibition of DENV production in Crispr-Cas9 GFP-LC3 knocked-in THP-1 monocytes. The core structure of Ka-003, which is a methyl cyclohexene derivative, resembles those found in mulberry plants, and could be synthetically prepared in a bioinspired fashion. Taken together, data indicate that Ka-003 hampered autophagy and limited DENV replication. The low cytotoxicity of Ka-003 suggests its therapeutic potential, which warrants further studies for the lead optimization of the compound for dengue treatment.<\/jats:p>","DOI":"10.3390\/v15102012","type":"journal-article","created":{"date-parts":[[2023,9,28]],"date-time":"2023-09-28T01:42:12Z","timestamp":1695865332000},"page":"2012","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Novel Potent Autophagy Inhibitor Ka-003 Inhibits Dengue Virus Replication"],"prefix":"10.3390","volume":"15","author":[{"given":"Jitra","family":"Limthongkul","sequence":"first","affiliation":[{"name":"Department of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kornkamon","family":"Akkarasereenon","sequence":"additional","affiliation":[{"name":"Laboratory of Medicinal Chemistry, Chulabhorn Research Institute, Bangkok 10210, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanpitcha","family":"Yodweerapong","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7907-1925","authenticated-orcid":false,"given":"Poramate","family":"Songthammawat","sequence":"additional","affiliation":[{"name":"Laboratory of Medicinal Chemistry, Chulabhorn Research Institute, Bangkok 10210, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pirut","family":"Tong-Ngam","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom 73170, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alisa","family":"Tubsuwan","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom 73170, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nawapol","family":"Kunkaew","sequence":"additional","affiliation":[{"name":"Mahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1046-7152","authenticated-orcid":false,"given":"Phongthon","family":"Kanjanasirirat","sequence":"additional","affiliation":[{"name":"Excellent Center for Drug Discovery, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"},{"name":"Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanawadee","family":"Khumpanied","sequence":"additional","affiliation":[{"name":"Excellent Center for Drug Discovery, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Warawuth","family":"Wannalo","sequence":"additional","affiliation":[{"name":"Excellent Center for Drug Discovery, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sukathida","family":"Ubol","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suparerk","family":"Borwornpinyo","sequence":"additional","affiliation":[{"name":"Excellent Center for Drug Discovery, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"},{"name":"Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2893-1598","authenticated-orcid":false,"given":"Poonsakdi","family":"Ploypradith","sequence":"additional","affiliation":[{"name":"Laboratory of Medicinal Chemistry, Chulabhorn Research Institute, Bangkok 10210, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marisa","family":"Ponpuak","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,9,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"16055","DOI":"10.1038\/nrdp.2016.55","article-title":"Dengue infection","volume":"2","author":"Guzman","year":"2016","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_2","unstructured":"WHO (2023). Dengue and Severe Dengue Fact Sheets, WHO."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"532","DOI":"10.1038\/nri3014","article-title":"Immunity to dengue virus: A tale of original antigenic sin and tropical cytokine storms","volume":"11","author":"Rothman","year":"2011","journal-title":"Nat. Reviews. Immunol."},{"key":"ref_4","first-page":"19839","article-title":"Dengue viruses\u2014An overview","volume":"3","author":"Back","year":"2013","journal-title":"Infect. Ecol. Epidemiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1445","DOI":"10.1007\/s00705-013-1645-3","article-title":"Secondary infection as a risk factor for dengue hemorrhagic fever\/dengue shock syndrome: An historical perspective and role of antibody-dependent enhancement of infection","volume":"158","author":"Guzman","year":"2013","journal-title":"Arch. Virol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1099\/vir.0.007914-0","article-title":"A role for autophagolysosomes in dengue virus 3 production in HepG2 cells","volume":"90","author":"Khakpoor","year":"2009","journal-title":"J. Gen. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1016\/j.virol.2008.02.016","article-title":"Autophagic machinery activated by dengue virus enhances virus replication","volume":"374","author":"Lee","year":"2008","journal-title":"Virology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1128\/JVI.02177-12","article-title":"Inhibition of cellular autophagy deranges dengue virion maturation","volume":"87","author":"Mateo","year":"2013","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"8026","DOI":"10.1128\/JVI.00787-15","article-title":"Dengue Virus Inhibition of Autophagic Flux and Dependency of Viral Replication on Proteasomal Degradation of the Autophagy Receptor p62","volume":"89","author":"Metz","year":"2015","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1099\/vir.0.005355-0","article-title":"Co-localization of constituents of the dengue virus translation and replication machinery with amphisomes","volume":"90","author":"Panyasrivanit","year":"2009","journal-title":"J. Gen. Virol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1016\/j.immuni.2021.01.018","article-title":"Autophagy in inflammation, infection, and immunometabolism","volume":"54","author":"Deretic","year":"2021","journal-title":"Immunity"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e108863","DOI":"10.15252\/embj.2021108863","article-title":"Autophagy in major human diseases","volume":"40","author":"Klionsky","year":"2021","journal-title":"EMBO J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1146\/annurev-pathol-020712-163918","article-title":"Autophagy as a stress-response and quality-control mechanism: Implications for cell injury and human disease","volume":"8","author":"Murrow","year":"2013","journal-title":"Annu. Rev. Pathol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1038\/nri3532","article-title":"Autophagy in infection, inflammation and immunity","volume":"13","author":"Deretic","year":"2013","journal-title":"Nat. Rev. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1038\/s41556-021-00669-y","article-title":"Autophagosome biogenesis comes out of the black box","volume":"23","author":"Chang","year":"2021","journal-title":"Nat. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1038\/cr.2013.168","article-title":"The machinery of macroautophagy","volume":"24","author":"Feng","year":"2014","journal-title":"Cell Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/j.ceb.2009.11.014","article-title":"Mammalian autophagy: Core molecular machinery and signaling regulation","volume":"22","author":"Yang","year":"2010","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Lee, Y.-R., Hu, H.-Y., Kuo, S.-H., Lei, H.-Y., Lin, Y.-S., Yeh, T.-M., Liu, C.-C., and Liu, H.-S. (2013). Dengue virus infection induces autophagy: An in vivo study. J. Biomed. Sci., 20.","DOI":"10.1186\/1423-0127-20-65"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1016\/j.chom.2010.10.006","article-title":"Dengue virus-induced autophagy regulates lipid metabolism","volume":"8","author":"Heaton","year":"2010","journal-title":"Cell Host Microbe"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"32243","DOI":"10.1038\/srep32243","article-title":"Autophagy-associated dengue vesicles promote viral transmission avoiding antibody neutralization","volume":"6","author":"Wu","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2281","DOI":"10.1038\/nprot.2013.143","article-title":"Genome engineering using the CRISPR-Cas9 system","volume":"8","author":"Ran","year":"2013","journal-title":"Nat. Protoc."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"13410","DOI":"10.1021\/acs.joc.9b01596","article-title":"Total Synthesis of Palodesangren B Trimethyl Ether and D Dimethyl Ether via a Late-Stage Formation of 2H-Pyran-2-one of the Tetrahydrobenzo[c]pyranochromenone Core","volume":"84","author":"Tangdenpaisal","year":"2019","journal-title":"J. Org. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"9177","DOI":"10.1038\/s41598-019-45641-x","article-title":"A novel potent autophagy inhibitor ECDD-S27 targets vacuolar ATPase and inhibits cancer cell survival","volume":"9","author":"Paha","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.antiviral.2014.03.014","article-title":"Amodiaquine, an antimalarial drug, inhibits dengue virus type 2 replication and infectivity","volume":"106","author":"Boonyasuppayakorn","year":"2014","journal-title":"Antivir. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"19963","DOI":"10.1038\/s41598-020-77003-3","article-title":"High-content screening of Thai medicinal plants reveals Boesenbergia rotunda extract and its component Panduratin A as anti-SARS-CoV-2 agents","volume":"10","author":"Kanjanasirirat","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/15548627.2020.1797280","article-title":"Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)","volume":"17","author":"Klionsky","year":"2021","journal-title":"Autophagy"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"5720","DOI":"10.1093\/emboj\/19.21.5720","article-title":"LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing","volume":"19","author":"Kabeya","year":"2000","journal-title":"EMBO J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"780","DOI":"10.4161\/auto.19385","article-title":"A role for Atg8-PE deconjugation in autophagosome biogenesis","volume":"8","author":"Nair","year":"2012","journal-title":"Autophagy"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"883","DOI":"10.4161\/auto.19652","article-title":"Dual roles of Atg8-PE deconjugation by Atg4 in autophagy","volume":"8","author":"Yu","year":"2012","journal-title":"Autophagy"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1083\/jcb.200507002","article-title":"p62\/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death","volume":"171","author":"Lamark","year":"2005","journal-title":"J. Cell Biol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Guo, H., Xu, Y., Huang, W., Zhou, H., Zheng, Z., Zhao, Y., He, B., Zhu, T., Tang, S., and Zhu, Q. (2016). Kuwanon G Preserves LPS-Induced Disruption of Gut Epithelial Barrier In Vitro. Molecules, 21.","DOI":"10.3390\/molecules21111597"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"425","DOI":"10.3892\/etm.2021.9869","article-title":"Kuwanon G protects HT22 cells from advanced glycation end product-induced damage","volume":"21","author":"Gan","year":"2021","journal-title":"Exp. Ther. Med."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Thabti, I., Albert, Q., Philippot, S., Dupire, F., Westerhuis, B., Fontanay, S., Risler, A., Kassab, T., Elfalleh, W., and Aferchichi, A. (2020). Advances on Antiviral Activity of Morus spp. Plant Extracts: Human Coronavirus and Virus-Related Respiratory Tract Infections in the Spotlight. Molecules, 25.","DOI":"10.3390\/molecules25081876"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Hsu, J.H., Yang, C.S., and Chen, J.J. (2022). Antioxidant, Anti-alpha-Glucosidase, Antityrosinase, and Anti-Inflammatory Activities of Bioactive Components from Morus alba. Antioxidants, 11.","DOI":"10.3390\/antiox11112222"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1667","DOI":"10.1248\/bpb.b16-00401","article-title":"Anti-HSV Activity of Kuwanon X from Mulberry Leaves with Genes Expression Inhibitory and HSV-1 Induced NF-kappaB Deactivated Properties","volume":"39","author":"Ma","year":"2016","journal-title":"Biol. Pharm. Bull."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"783","DOI":"10.4269\/ajtmh.18-0761","article-title":"Significance of Autophagy in Dengue Virus Infection: A Brief Review","volume":"100","author":"Acharya","year":"2019","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e2127","DOI":"10.1038\/cddis.2015.409","article-title":"Dengue-induced autophagy, virus replication and protection from cell death require ER stress (PERK) pathway activation","volume":"7","author":"Datan","year":"2016","journal-title":"Cell Death Dis."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"4153","DOI":"10.1073\/pnas.0913860107","article-title":"ATM signals to TSC2 in the cytoplasm to regulate mTORC1 in response to ROS","volume":"107","author":"Alexander","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e02020-16","DOI":"10.1128\/JVI.02020-16","article-title":"Dengue Virus Activates the AMP Kinase-mTOR Axis To Stimulate a Proviral Lipophagy","volume":"91","author":"Jordan","year":"2017","journal-title":"J. Virol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1749","DOI":"10.1038\/sj.emboj.7601623","article-title":"Reactive oxygen species are essential for autophagy and specifically regulate the activity of Atg4","volume":"26","author":"Shvets","year":"2007","journal-title":"EMBO J."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/10\/2012\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:59:54Z","timestamp":1760129994000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/10\/2012"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,27]]},"references-count":40,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2023,10]]}},"alternative-id":["v15102012"],"URL":"https:\/\/doi.org\/10.3390\/v15102012","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,27]]}}}