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Policresulen, a novel NS2B/NS3 protease inhibitor, effectively inhibits the replication of DENV2 virus in BHK-21 cells.

Wu DW, Mao F, Ye Y, Li J, Xu CL, Luo XM, Chen J, Shen X - Acta Pharmacol. Sin. (2015)

Bottom Line: In our in-house library of old drugs (~1000 compounds), a topical hemostatic and antiseptic 2-hydroxy-3,5-bis[(4-hydroxy-2-methyl-5-sulfophenyl)methyl]-4-methyl-benzene-sulfonic acid (policresulen) was found to be a potent inhibitor of DENV2 NS2B/NS3 protease with IC50 of 0.48 μg/mL.Policresulen is a potent inhibitor of DENV2 NS2B/NS3 protease that inhibits DENV2 replication in BHK-21 cells.The binding mode of the protease and policresulen provides useful hints for designing new type of inhibitors against the protease.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China.

ABSTRACT

Aim: Dengue is a severe epidemic disease caused by dengue virus (DENV) infection, for which no effective treatment is available. The protease complex, consisting of nonstructural protein 3 (NS3) and its cofactor NS2B, plays a pivotal role in the replication of DENV, thus may be a potential target for anti-DENV drugs. Here, we report a novel inhibitor of DENV2 NS2B/NS3 protease and its antiviral action.

Methods: An enzymatic inhibition assay was used for screening DENV2 NS2B/NS3 inhibitors. Cytotoxicity to BHK-21 cells was assessed with MTT assay. Antiviral activity was evaluated in BHK-21 cells transfected with Rlu-DENV-Rep. The molecular mechanisms of the antiviral action was analyzed using surface plasmon resonance, ultraviolet-visible spectral analysis and differential scanning calorimetry assays, as well as molecular docking analysis combined with site-directed mutagenesis.

Results: In our in-house library of old drugs (~1000 compounds), a topical hemostatic and antiseptic 2-hydroxy-3,5-bis[(4-hydroxy-2-methyl-5-sulfophenyl)methyl]-4-methyl-benzene-sulfonic acid (policresulen) was found to be a potent inhibitor of DENV2 NS2B/NS3 protease with IC50 of 0.48 μg/mL. Furthermore, policresulen inhibited DENV2 replication in BHK-21 cells with IC50 of 4.99 μg/mL, whereas its IC50 for cytotoxicity to BHK-21 cells was 459.45 μg/mL. Policresulen acted as a competitive inhibitor of the protease, and slightly affected the protease stability. Using biophysical technology-based assays and molecular docking analysis combined with site-directed mutagenesis, we demonstrated that the residues Gln106 and Arg133 of DENV2 NS2B/NS3 protease directly interacted with policresulen via hydrogen bonding.

Conclusion: Policresulen is a potent inhibitor of DENV2 NS2B/NS3 protease that inhibits DENV2 replication in BHK-21 cells. The binding mode of the protease and policresulen provides useful hints for designing new type of inhibitors against the protease.

No MeSH data available.


Related in: MedlinePlus

Policresulen functioned as an inhibitor of DENV2 NS2B/NS3 protease. (A) Chemical structure of policresulen. (B) The dose-dependent inhibitory effect of policresulen against DENV2 NS2B/NS3 protease. (C) The cytotoxicity of policresulen was evaluated by MTT assay. BHK-21 cells were treated with different concentrations of policresulen for 48 h.
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fig1: Policresulen functioned as an inhibitor of DENV2 NS2B/NS3 protease. (A) Chemical structure of policresulen. (B) The dose-dependent inhibitory effect of policresulen against DENV2 NS2B/NS3 protease. (C) The cytotoxicity of policresulen was evaluated by MTT assay. BHK-21 cells were treated with different concentrations of policresulen for 48 h.

Mentions: In the current work, we performed an enzymatic inhibition assay to discover inhibitors of this recombinant DENV2 NS2B/NS3 protease in vitro against our in-house library of old drugs, and 2-hydroxy-3,5-bis[(4-hydroxy-2-methyl-5-sulfo-phenyl)methyl]-4-methyl-benzenesulfonic acid (policresulen, Figure 1A), a clinical medication used for debridement and antimicrobial applications in gynecology29, was finally discovered to be a novel inhibitor of this protease. The subsequent discovery of its antiviral properties demonstrated that policresulen could efficiently reduce the replication of DENV2 in BHK-21 cells. Additionally, the molecular mechanism of policresulen against the DENV2 NS2B/NS3 protease was further investigated by binding affinity, thermal stability and molecular docking assays. Our results have demonstrated that policresulen might have potential in the treatment of DENV infection, and the expounded binding pocket of policresulen for this protease may provide useful data for designing new inhibitors against DENV.


Policresulen, a novel NS2B/NS3 protease inhibitor, effectively inhibits the replication of DENV2 virus in BHK-21 cells.

Wu DW, Mao F, Ye Y, Li J, Xu CL, Luo XM, Chen J, Shen X - Acta Pharmacol. Sin. (2015)

Policresulen functioned as an inhibitor of DENV2 NS2B/NS3 protease. (A) Chemical structure of policresulen. (B) The dose-dependent inhibitory effect of policresulen against DENV2 NS2B/NS3 protease. (C) The cytotoxicity of policresulen was evaluated by MTT assay. BHK-21 cells were treated with different concentrations of policresulen for 48 h.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4561973&req=5

fig1: Policresulen functioned as an inhibitor of DENV2 NS2B/NS3 protease. (A) Chemical structure of policresulen. (B) The dose-dependent inhibitory effect of policresulen against DENV2 NS2B/NS3 protease. (C) The cytotoxicity of policresulen was evaluated by MTT assay. BHK-21 cells were treated with different concentrations of policresulen for 48 h.
Mentions: In the current work, we performed an enzymatic inhibition assay to discover inhibitors of this recombinant DENV2 NS2B/NS3 protease in vitro against our in-house library of old drugs, and 2-hydroxy-3,5-bis[(4-hydroxy-2-methyl-5-sulfo-phenyl)methyl]-4-methyl-benzenesulfonic acid (policresulen, Figure 1A), a clinical medication used for debridement and antimicrobial applications in gynecology29, was finally discovered to be a novel inhibitor of this protease. The subsequent discovery of its antiviral properties demonstrated that policresulen could efficiently reduce the replication of DENV2 in BHK-21 cells. Additionally, the molecular mechanism of policresulen against the DENV2 NS2B/NS3 protease was further investigated by binding affinity, thermal stability and molecular docking assays. Our results have demonstrated that policresulen might have potential in the treatment of DENV infection, and the expounded binding pocket of policresulen for this protease may provide useful data for designing new inhibitors against DENV.

Bottom Line: In our in-house library of old drugs (~1000 compounds), a topical hemostatic and antiseptic 2-hydroxy-3,5-bis[(4-hydroxy-2-methyl-5-sulfophenyl)methyl]-4-methyl-benzene-sulfonic acid (policresulen) was found to be a potent inhibitor of DENV2 NS2B/NS3 protease with IC50 of 0.48 μg/mL.Policresulen is a potent inhibitor of DENV2 NS2B/NS3 protease that inhibits DENV2 replication in BHK-21 cells.The binding mode of the protease and policresulen provides useful hints for designing new type of inhibitors against the protease.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China.

ABSTRACT

Aim: Dengue is a severe epidemic disease caused by dengue virus (DENV) infection, for which no effective treatment is available. The protease complex, consisting of nonstructural protein 3 (NS3) and its cofactor NS2B, plays a pivotal role in the replication of DENV, thus may be a potential target for anti-DENV drugs. Here, we report a novel inhibitor of DENV2 NS2B/NS3 protease and its antiviral action.

Methods: An enzymatic inhibition assay was used for screening DENV2 NS2B/NS3 inhibitors. Cytotoxicity to BHK-21 cells was assessed with MTT assay. Antiviral activity was evaluated in BHK-21 cells transfected with Rlu-DENV-Rep. The molecular mechanisms of the antiviral action was analyzed using surface plasmon resonance, ultraviolet-visible spectral analysis and differential scanning calorimetry assays, as well as molecular docking analysis combined with site-directed mutagenesis.

Results: In our in-house library of old drugs (~1000 compounds), a topical hemostatic and antiseptic 2-hydroxy-3,5-bis[(4-hydroxy-2-methyl-5-sulfophenyl)methyl]-4-methyl-benzene-sulfonic acid (policresulen) was found to be a potent inhibitor of DENV2 NS2B/NS3 protease with IC50 of 0.48 μg/mL. Furthermore, policresulen inhibited DENV2 replication in BHK-21 cells with IC50 of 4.99 μg/mL, whereas its IC50 for cytotoxicity to BHK-21 cells was 459.45 μg/mL. Policresulen acted as a competitive inhibitor of the protease, and slightly affected the protease stability. Using biophysical technology-based assays and molecular docking analysis combined with site-directed mutagenesis, we demonstrated that the residues Gln106 and Arg133 of DENV2 NS2B/NS3 protease directly interacted with policresulen via hydrogen bonding.

Conclusion: Policresulen is a potent inhibitor of DENV2 NS2B/NS3 protease that inhibits DENV2 replication in BHK-21 cells. The binding mode of the protease and policresulen provides useful hints for designing new type of inhibitors against the protease.

No MeSH data available.


Related in: MedlinePlus