Limits...
Antiviral Potential of a Novel Compound CW-33 against Enterovirus A71 via Inhibition of Viral 2A Protease.

Wang CY, Huang AC, Hour MJ, Huang SH, Kung SH, Chen CH, Chen IC, Chang YS, Lien JC, Lin CW - Viruses (2015)

Bottom Line: 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).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.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 40402, Taiwan. yschang@mail.cmu.edu.tw.

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.

No MeSH data available.


Related in: MedlinePlus

Inhibition of EV-A71 plaque formation by CW-33 or IFN-β alone. Monolayer of RD cells in 6-well plates were infected with EV-A71 (100 pfu), and then immediately treated with indicated concentrations of CW-33 (A) or IFN-β (C). After 1-h absorption, cell monolayer was washed with PBS, then overlaid with medium containing 1.5% agar. After 48-h incubation, plaque number was counted after staining by 0.1% Crystal Violet, Inhibitory activities of CW-33 (B) or IFN-β (D) were calculated from ratio of experimental data to mock control. *** p value < 0.001 by Scheffe test.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4488731&req=5

viruses-07-02764-f003: Inhibition of EV-A71 plaque formation by CW-33 or IFN-β alone. Monolayer of RD cells in 6-well plates were infected with EV-A71 (100 pfu), and then immediately treated with indicated concentrations of CW-33 (A) or IFN-β (C). After 1-h absorption, cell monolayer was washed with PBS, then overlaid with medium containing 1.5% agar. After 48-h incubation, plaque number was counted after staining by 0.1% Crystal Violet, Inhibitory activities of CW-33 (B) or IFN-β (D) were calculated from ratio of experimental data to mock control. *** p value < 0.001 by Scheffe test.

Mentions: Cytotoxicity of CW-33 to RD cells was initially assessed using MTT assay (Figure 2A). Survival rate exceeded 50% when cells treated with high concentration of CW-33 at 1000 μM, proving CW-33 definitely less cytotoxic. Antiviral activity of CW-33 against EV-A71 was later tested by cytopathic effect inhibition and plaque reduction assay (Figure 2B,C and Figure 3A,B). CW-33 concentration-dependently suppressed EV-A71-induced cytopathic effect in RD cells (Figure 2B), as well as reducing apoptotic rate of virus-infected cells (Figure 2C) (p < 0.001). CW-33 likewise showed plaque reduction activity with IC50 of 193.8 μM. The results indicated CW-33 displaying a moderate antiviral activity against EV-A71.


Antiviral Potential of a Novel Compound CW-33 against Enterovirus A71 via Inhibition of Viral 2A Protease.

Wang CY, Huang AC, Hour MJ, Huang SH, Kung SH, Chen CH, Chen IC, Chang YS, Lien JC, Lin CW - Viruses (2015)

Inhibition of EV-A71 plaque formation by CW-33 or IFN-β alone. Monolayer of RD cells in 6-well plates were infected with EV-A71 (100 pfu), and then immediately treated with indicated concentrations of CW-33 (A) or IFN-β (C). After 1-h absorption, cell monolayer was washed with PBS, then overlaid with medium containing 1.5% agar. After 48-h incubation, plaque number was counted after staining by 0.1% Crystal Violet, Inhibitory activities of CW-33 (B) or IFN-β (D) were calculated from ratio of experimental data to mock control. *** p value < 0.001 by Scheffe test.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488731&req=5

viruses-07-02764-f003: Inhibition of EV-A71 plaque formation by CW-33 or IFN-β alone. Monolayer of RD cells in 6-well plates were infected with EV-A71 (100 pfu), and then immediately treated with indicated concentrations of CW-33 (A) or IFN-β (C). After 1-h absorption, cell monolayer was washed with PBS, then overlaid with medium containing 1.5% agar. After 48-h incubation, plaque number was counted after staining by 0.1% Crystal Violet, Inhibitory activities of CW-33 (B) or IFN-β (D) were calculated from ratio of experimental data to mock control. *** p value < 0.001 by Scheffe test.
Mentions: Cytotoxicity of CW-33 to RD cells was initially assessed using MTT assay (Figure 2A). Survival rate exceeded 50% when cells treated with high concentration of CW-33 at 1000 μM, proving CW-33 definitely less cytotoxic. Antiviral activity of CW-33 against EV-A71 was later tested by cytopathic effect inhibition and plaque reduction assay (Figure 2B,C and Figure 3A,B). CW-33 concentration-dependently suppressed EV-A71-induced cytopathic effect in RD cells (Figure 2B), as well as reducing apoptotic rate of virus-infected cells (Figure 2C) (p < 0.001). CW-33 likewise showed plaque reduction activity with IC50 of 193.8 μM. The results indicated CW-33 displaying a moderate antiviral activity against EV-A71.

Bottom Line: 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).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.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 40402, Taiwan. yschang@mail.cmu.edu.tw.

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.

No MeSH data available.


Related in: MedlinePlus