title: Antiviral Potential of a Novel Compound CW-33 against Enterovirus A71 via Inhibition of Viral 2A Protease.
abstract: Enterovirus A71 (EV-A71) in the Picornaviridae family causes hand-foot-and-mouth disease, aseptic meningitis, severe central nervous system disease, even death. EV-A71 2A protease cleaves Type I interferon (IFN)-α/β receptor 1 (IFNAR1) to block IFN-induced Jak/STAT signaling. This study investigated anti-EV-A7l activity and synergistic mechanism(s) of a novel furoquinoline alkaloid compound CW-33 alone and in combination with IFN-β Anti-EV-A71 activities of CW-33 alone and in combination with IFN-β were evaluated by inhibitory assays of virus-induced apoptosis, plaque formation, and virus yield. CW-33 showed antiviral activities with an IC50 of near 200 µM in EV-A71 plaque reduction and virus yield inhibition assays. While, anti-EV-A71 activities of CW-33 combined with 100 U/mL IFN-β exhibited a synergistic potency with an IC50 of approximate 1 µM in plaque reduction and virus yield inhibition assays. Molecular docking revealed CW-33 binding to EV-A71 2A protease active sites, correlating with an inhibitory effect of CW33 on in vitro enzymatic activity of recombinant 2A protease IC50 = 53.1 µM). Western blotting demonstrated CW-33 specifically inhibiting 2A protease-mediated cleavage of IFNAR1. CW-33 also recovered Type I IFN-induced Tyk2 and STAT1 phosphorylation as well as 2\',5\'-OAS upregulation in EV-A71 infected cells. The results demonstrated CW-33 inhibiting viral 2A protease activity to reduce Type I IFN antagonism of EV-A71. Therefore, CW-33 combined with a low-dose of Type I IFN could be applied in developing alternative approaches to treat EV-A71 infection.
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 40402, Taiwan. yschang@mail.cmu.edu.tw.|School of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, Taichung 40402, Taiwan. yschang@mail.cmu.edu.tw.
authors.forename: Ching-Ying
authors.initials: CY
authors.lastname: Wang
authors.affiliation: Department of Nursing, St. Mary\'s Junior College of Medicine, Nursing and Management, Yilan County 266, Taiwan. haj@smc.edu.tw.
authors.forename: An-Cheng
authors.initials: AC
authors.lastname: Huang
authors.affiliation: School of Pharmacy, China Medical University, Taichung 40402, Taiwan. mjhou@mail.cmu.edu.tw.
authors.forename: Mann-Jen
authors.initials: MJ
authors.lastname: Hour
authors.affiliation: Department of Biotechnology, Asia University, Wufeng, Taichung 41354, Taiwan. shhuang@asia.edu.tw.
authors.forename: Su-Hua
authors.initials: SH
authors.lastname: Huang
authors.affiliation: Department of Biotechnology and Laboratory Science in Medicine, National Yang Ming University, Taipei 11221, Taiwan. szkung@ym.edu.tw.
authors.forename: Szu-Hao
authors.initials: SH
authors.lastname: Kung
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 40402, Taiwan. chchen@mail.cmu.edu.tw.
authors.forename: Chao-Hsien
authors.initials: CH
authors.lastname: Chen
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 40402, Taiwan. hiichieh@gmail.com.
authors.forename: I-Chieh
authors.initials: IC
authors.lastname: Chen
authors.affiliation: School of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, Taichung 40402, Taiwan. yschang@mail.cmu.edu.tw.
authors.forename: Yuan-Shiun
authors.initials: YS
authors.lastname: Chang
authors.affiliation: School of Pharmacy, China Medical University, Taichung 40402, Taiwan. jclien@mail.cmu.edu.tw.
authors.forename: Jin-Cherng
authors.initials: JC
authors.lastname: Lien
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 40402, Taiwan. cwlin@mail.cmu.edu.tw.|Department of Biotechnology, Asia University, Wufeng, Taichung 41354, Taiwan. cwlin@mail.cmu.edu.tw.
authors.forename: Cheng-Wen
authors.initials: CW
authors.lastname: Lin
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D015153:Blotting, Western; D003546:Cysteine Endopeptidases; D004357:Drug Synergism; D029821:Enterovirus A, Human / Q000187:drug effects* / Q000201:enzymology*; D006801:Humans; D020128:Inhibitory Concentration 50; D016899:Interferon-beta / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D062105:Molecular Docking Simulation; D011480:Protease Inhibitors / Q000494:pharmacology*; D011485:Protein Binding; D059748:Proteolysis / Q000187:drug effects; D015363:Quinolones / Q000494:pharmacology; D053633:Receptor, Interferon alpha-beta / Q000378:metabolism; D019562:Viral Load; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000037:antagonists & inhibitors*
chemical_list: D000998:Antiviral Agents; C507964:IFNAR1 protein, human; D011480:Protease Inhibitors; D015363:Quinolones; D014764:Viral Proteins; D053633:Receptor, Interferon alpha-beta; D016899:Interferon-beta; D003546:Cysteine Endopeptidases; C057400:picornain 2A, Picornavirus
keywords: 2A protease; antagonism; enterovirus A71; inhibitor; type I interferon