title: Antiviral activities of niclosamide and nitazoxanide against chikungunya virus entry and transmission.
abstract: Chikungunya disease results from an infection with the arbovirus, chikungunya virus (CHIKV). Symptoms of CHIKV include fever and persistent, severe arthritis. In recent years, several antiviral drugs have been evaluated in clinical trials; however, no registered antivirals have been approved for clinical therapy. In this study, we established a high-throughput screening (HTS) system based on CHIKV 26S mediated insect cell fusion inhibition assay. Our screening system was able to search potential anti-CHIKV drugs in vitro. Using this system, four compounds (niclosamide, nitazoxanide, niflumic acid, tolfenamic acid) were identified. These compounds were then further analyzed using a microneutralization assay. We determined that niclosamide and nitazoxanide exhibit ability to against CHIKV-induced CPE. The anti-CHIKV abilities of these compounds were further confirmed by RT-qPCR and IFA. Moreover, niclosamide and nitazoxanide were found to (1) limit virus entry, (2) inhibit both viral release and cell-to-cell transmission, and (3) possess broad anti-alphavius activities, including against two clinical CHIKV isolates and two alphaviruses: Sindbis virus (SINV) and Semliki forest virus (SFV). In conclusion, our findings suggested that niclosamide and nitazoxanide were able to inhibit CHIKV entry and transmission, which might provide a basis for the development of novel human drug therapies against CHIKV and other alphavirus infections.
authors.affiliation: Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, ROC.
authors.forename: Yu-Ming
authors.initials: YM
authors.lastname: Wang
authors.affiliation: Department of Clinical Laboratory Sciences and Medical Biotechnology, National Taiwan University, Taipei, Taiwan, ROC.
authors.forename: Jeng-Wei
authors.initials: JW
authors.lastname: Lu
authors.affiliation: Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, ROC; Department and Graduate Institute of Microbiology and Immunology, National Defense Medical Center, Taipei, Taiwan, ROC.
authors.forename: Chang-Chi
authors.initials: CC
authors.lastname: Lin
authors.affiliation: Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, ROC.
authors.forename: Yuan-Fan
authors.initials: YF
authors.lastname: Chin
authors.affiliation: Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li, Taiwan, ROC; Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, Taiwan, ROC.
authors.forename: Tzong-Yuan
authors.initials: TY
authors.lastname: Wu
authors.affiliation: Department of Clinical Laboratory Sciences and Medical Biotechnology, National Taiwan University, Taipei, Taiwan, ROC; Department of Laboratory Medicine, National Taiwan University Hospital, Taipei, Taiwan, ROC.
authors.forename: Liang-In
authors.initials: LI
authors.lastname: Lin
authors.affiliation: Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, ROC.
authors.forename: Zheng-Zong
authors.initials: ZZ
authors.lastname: Lai
authors.affiliation: Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, ROC; Department and Graduate Institute of Microbiology and Immunology, National Defense Medical Center, Taipei, Taiwan, ROC. Electronic address: szucheng1234@gmail.com.
authors.forename: Szu-Cheng
authors.initials: SC
authors.lastname: Kuo
authors.affiliation: Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, ROC; School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, ROC. Electronic address: ejung330@gmail.com.
authors.forename: Yi-Jung
authors.initials: YJ
authors.lastname: Ho
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D065632:Chikungunya Fever / Q000635:transmission / Q000821:virology; D002646:Chikungunya virus / Q000187:drug effects* / Q000502:physiology; D055808:Drug Discovery; D057166:High-Throughput Screening Assays; D006801:Humans; D009534:Niclosamide / Q000494:pharmacology*; D009574:Nitro Compounds; D012672:Semliki forest virus / Q000187:drug effects; D012845:Sindbis Virus / Q000187:drug effects; D013844:Thiazoles / Q000494:pharmacology*; D053586:Virus Internalization / Q000187:drug effects*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D009574:Nitro Compounds; D013844:Thiazoles; D009534:Niclosamide; C041747:nitazoxanide
keywords: Antiviral screening; Chikungunya virus; Insect cell fusion assay; Niclosamide; Nitazoxanide