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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

CW-33 chemical synthesis and NMR characterization (Ethyl 2-(3′,5′-dimethylanilino)-4-oxo-4,5-dihydrofuran-3-carboxylate). Flow diagram of CW-33 synthesis appears in (A). 1H-NMR and 13C-NMR spectra of CW-33 are shown in (B,C), respectively.
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viruses-07-02764-f001: CW-33 chemical synthesis and NMR characterization (Ethyl 2-(3′,5′-dimethylanilino)-4-oxo-4,5-dihydrofuran-3-carboxylate). Flow diagram of CW-33 synthesis appears in (A). 1H-NMR and 13C-NMR spectra of CW-33 are shown in (B,C), respectively.

Mentions: Furoquinoline alkaloids are bioactive compounds in many plants in the Rutaceae family, such as Hortia oreadica, H. apiculata, Teclea afzelii, Oricia suaveolens, and Balfourodendron riedelianum [18]. Most furoquinoline alkaloids possess many biological activities: e.g., antifungal [19], antimicrobial [20], antioxidant [21], and anticancer activities [22]. Several synthesized compounds based on furoquinoline skeleton, such as n-alkyl-2,3,4,9-tetrahydrofuro [2,3-b] quinoline-3,4-diones and N-benzyl-7-methoxy-2,3,4,9-tetrahydrofuro[2,3-b]quinoline-3,4-dione (HA-7), exhibit anti-inflammatory, antiallergic and antiarrhythmic activities [23,24]. Compound CW-33 is a novel synthetic derivative of furoquinoline alkaloid; flow diagram of CW-33 synthesis appears in Figure 1A. This study rates anti-EV-A7l activity of a novel furoquinoline alkaloid compound CW-33 alone and in combination with IFNβ. CW-33 shows an inhibitory effect on EV-A71 replication in vitro. Combined treatment with CW-33 and IFN-β exhibits a synergistic antiviral effect on reducing EV-A71-induced cytopathy (apoptosis), virus yield, and plaque formation. Molecular docking analysis indicated the binding of CW-33 to EV-A71 2A protease active sites, confirmed by the inhibitory effect of CW-33 on recombinant 2A enzymatic activity as well as recovering the protein levels of IFNAR1 in EV-A71-infected cells.


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)

CW-33 chemical synthesis and NMR characterization (Ethyl 2-(3′,5′-dimethylanilino)-4-oxo-4,5-dihydrofuran-3-carboxylate). Flow diagram of CW-33 synthesis appears in (A). 1H-NMR and 13C-NMR spectra of CW-33 are shown in (B,C), respectively.
© Copyright Policy
Related In: Results  -  Collection

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

viruses-07-02764-f001: CW-33 chemical synthesis and NMR characterization (Ethyl 2-(3′,5′-dimethylanilino)-4-oxo-4,5-dihydrofuran-3-carboxylate). Flow diagram of CW-33 synthesis appears in (A). 1H-NMR and 13C-NMR spectra of CW-33 are shown in (B,C), respectively.
Mentions: Furoquinoline alkaloids are bioactive compounds in many plants in the Rutaceae family, such as Hortia oreadica, H. apiculata, Teclea afzelii, Oricia suaveolens, and Balfourodendron riedelianum [18]. Most furoquinoline alkaloids possess many biological activities: e.g., antifungal [19], antimicrobial [20], antioxidant [21], and anticancer activities [22]. Several synthesized compounds based on furoquinoline skeleton, such as n-alkyl-2,3,4,9-tetrahydrofuro [2,3-b] quinoline-3,4-diones and N-benzyl-7-methoxy-2,3,4,9-tetrahydrofuro[2,3-b]quinoline-3,4-dione (HA-7), exhibit anti-inflammatory, antiallergic and antiarrhythmic activities [23,24]. Compound CW-33 is a novel synthetic derivative of furoquinoline alkaloid; flow diagram of CW-33 synthesis appears in Figure 1A. This study rates anti-EV-A7l activity of a novel furoquinoline alkaloid compound CW-33 alone and in combination with IFNβ. CW-33 shows an inhibitory effect on EV-A71 replication in vitro. Combined treatment with CW-33 and IFN-β exhibits a synergistic antiviral effect on reducing EV-A71-induced cytopathy (apoptosis), virus yield, and plaque formation. Molecular docking analysis indicated the binding of CW-33 to EV-A71 2A protease active sites, confirmed by the inhibitory effect of CW-33 on recombinant 2A enzymatic activity as well as recovering the protein levels of IFNAR1 in EV-A71-infected cells.

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