Tags

Type your tag names separated by a space and hit enter

Policresulen, a novel NS2B/NS3 protease inhibitor, effectively inhibits the replication of DENV2 virus in BHK-21 cells.
Acta Pharmacol Sin. 2015 Sep; 36(9):1126-36.AP

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.

Links

PMC Free PDF

Authors+Show Affiliations

Wu DW
College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China.
Mao F
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
Ye Y
CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Li J
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
Xu CL
College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China.
Luo XM
CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Chen J
CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Shen X
CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.

MeSH

AnimalsAntiviral AgentsCell LineCresolsCricetinaeDengueDengue VirusDrug CombinationsFormaldehydeHumansMolecular Docking SimulationProtease InhibitorsViral Nonstructural ProteinsVirus Replication

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

26279156

Citation

Wu, Deng-wei, et al. "Policresulen, a Novel NS2B/NS3 Protease Inhibitor, Effectively Inhibits the Replication of DENV2 Virus in BHK-21 Cells." Acta Pharmacologica Sinica, vol. 36, no. 9, 2015, pp. 1126-36.
Wu DW, Mao F, Ye Y, et al. Policresulen, a novel NS2B/NS3 protease inhibitor, effectively inhibits the replication of DENV2 virus in BHK-21 cells. Acta Pharmacol Sin. 2015;36(9):1126-36.
Wu, D. W., Mao, F., Ye, Y., Li, J., Xu, C. L., Luo, X. M., Chen, J., & Shen, X. (2015). Policresulen, a novel NS2B/NS3 protease inhibitor, effectively inhibits the replication of DENV2 virus in BHK-21 cells. Acta Pharmacologica Sinica, 36(9), 1126-36. https://doi.org/10.1038/aps.2015.56
Wu DW, et al. Policresulen, a Novel NS2B/NS3 Protease Inhibitor, Effectively Inhibits the Replication of DENV2 Virus in BHK-21 Cells. Acta Pharmacol Sin. 2015;36(9):1126-36. PubMed PMID: 26279156.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Policresulen, a novel NS2B/NS3 protease inhibitor, effectively inhibits the replication of DENV2 virus in BHK-21 cells. AU - Wu,Deng-wei, AU - Mao,Fei, AU - Ye,Yan, AU - Li,Jian, AU - Xu,Chuan-lian, AU - Luo,Xiao-min, AU - Chen,Jing, AU - Shen,Xu, Y1 - 2015/08/17/ PY - 2015/02/02/received PY - 2015/05/08/accepted PY - 2015/8/18/entrez PY - 2015/8/19/pubmed PY - 2016/7/23/medline SP - 1126 EP - 36 JF - Acta pharmacologica Sinica JO - Acta Pharmacol Sin VL - 36 IS - 9 N2 - 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. SN - 1745-7254 UR - https://www.unboundmedicine.com/medline/citation/26279156/Policresulen_a_novel_NS2B/NS3_protease_inhibitor_effectively_inhibits_the_replication_of_DENV2_virus_in_BHK_21_cells_ DB - PRIME DP - Unbound Medicine ER -