title: Antiviral Activity of a Novel Compound CW-33 against Japanese Encephalitis Virus through Inhibiting Intracellular Calcium Overload.
abstract: Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, has five genotypes (I, II, III, IV, and V). JEV genotype I circulates widely in some Asian countries. However, current JEV vaccines based on genotype III strains show low neutralizing capacities against genotype I variants. In addition, JE has no specific treatment, except a few supportive treatments. Compound CW-33, an intermediate synthesized derivative of furoquinolines, was investigated for its antiviral activities against JEV in this study. CW-33 exhibited the less cytotoxicity to Syrian baby hamster kidney (BHK-21) and human medulloblastoma (TE761) cells. CW-33 dose-dependently reduced the cytopathic effect and apoptosis of JEV-infected cells. Supernatant virus yield assay pinpointed CW-33 as having potential anti-JEV activity with IC50 values ranging from 12.7 to 38.5 μM. Time-of-addition assay with CW-33 indicated that simultaneous and post-treatment had no plaque reduction activity, but continuous and simultaneous treatments proved to have highly effective antiviral activity, with IC50 values of 32.7 and 48.5 μM, respectively. CW-33 significantly moderated JEV-triggered Ca(2+) overload, which correlated with the recovery of mitochondria membrane potential as well as the activation of Akt/mTOR and Jak/STAT1 signals in treated infected cells. Phosphopeptide profiling by LC-MS/MS revealed that CW-33 upregulated proteins from the enzyme modulator category, such as protein phosphatase inhibitor 2 (I-2), Rho GTPase-activating protein 35, ARF GTPase-activating protein GIT2, and putative 3-phosphoinositide-dependent protein kinase 2. These enzyme modulators identified were associated with the activation of Akt/mTOR and Jak/STAT1 signals. Meanwhile, I-2 treatment substantially inhibited the apoptosis of JEV-infected cells. The results demonstrated that CW-33 exhibited a significant potential in the development of anti-JEV agents.
authors: Department of Biotechnology, Asia University, Wufeng, Taichung 413, Taiwan. shhuang@asia.edu.tw.
authors: Su-Hua
authors: SH
authors: Huang
authors: School of Pharmacy, China Medical University, Taichung 404, Taiwan. jclien@mail.cmu.edu.tw.
authors: Jin-Cherng
authors: JC
authors: Lien
authors: Proteomics Core Laboratory, Department of Medical Research, China Medical University Hospital, Taichung 404, Taiwan. cjchen@mail.cmu.edu.tw.
authors: Chao-Jung
authors: CJ
authors: Chen
authors: Proteomics Core Laboratory, Department of Medical Research, China Medical University Hospital, Taichung 404, Taiwan. yuhchyan1104@yahoo.com.tw.
authors: Yu-Ching
authors: YC
authors: Liu
authors: Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 404, Taiwan. spirit1126@hotmail.com.
authors: Ching-Ying
authors: CY
authors: Wang
authors: Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 404, Taiwan. anniepyng@hotmail.com.
authors: Chia-Fong
authors: CF
authors: Ping
authors: Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 404, Taiwan. wnedy55@yahoo.com.tw.
authors: Yu-Fong
authors: YF
authors: Lin
authors: Department of Nursing, St. Mary's Junior College of Medicine, Nursing and Management, Yilan County 266, Taiwan. haj@smc.edu.tw.
authors: An-Cheng
authors: AC
authors: Huang
authors: Department of Biotechnology, Asia University, Wufeng, Taichung 413, Taiwan. cwlin@mail.cmu.edu.tw.|Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 404, Taiwan. cwlin@mail.cmu.edu.tw.
authors: Cheng-Wen
authors: CW
authors: Lin
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D045744:Cell Line, Tumor; D006224:Cricetinae; D004664:Encephalitis Virus, Japanese / Q000187:drug effects* / Q000378:metabolism*; D020690:GTPase-Activating Proteins; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008647:Mesocricetus; D011804:Quinolines / Q000737:chemistry / Q000494:pharmacology; D050794:STAT1 Transcription Factor / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D020690:GTPase-Activating Proteins; D011804:Quinolines; D050794:STAT1 Transcription Factor; C085379:rho GTPase-activating protein; D002118:Calcium
keywords: Akt/mTOR; Ca2+ overload; Jak/STAT1; Japanese encephalitis virus; furoquinolines