文献类型：期刊文献

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

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

作者：Deng-wei WU[1];Fei MAO[2];Yan YE[3];Jian LI[2];Chuan-lian XU[1];Xiao-min LUO[3];Jing CHEN[3];Xu SHEN[3]

机构：[1]College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China;[2]Shanghai Key Laboratory of New Drug Design,School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China;[3]CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China

年份：2015

卷号：36

期号：9

起止页码：1126

中文期刊名：Acta Pharmacologica Sinica

外文期刊名：中国药理学报（英文版）

收录：CSTPCD;;Scopus;CSCD:【CSCD2015_2016】；PubMed;

基金：We gratefully acknowledge financial support from the National Natural Science Foundation of China （Grant No 81220108025, 21222211, 81373461, 91413102, 81473141, and 91213306）.

语种：英文

中文关键词：Dengue virus; antiviral drug; NS2B/NS3 protease; policresulen; molecular docking; molecular modeling

外文关键词：Dengue virus; antiviral drug; NS2B/NS3 protease; policresulen; molecular docking; molecular modeling

摘要：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-DENY drugs. Here, we report a novel inhibitor of DENY2 NS2B/NS3 protease and its antiviral action. Methods： An enzymatic inhibition assay was used for screening DENY2 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 （NIO00 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 ICso of 0.48 pg/mL. Furthermore, policresulen inhibited DENV2 replication in BHK-21 cells with ICso of 4,99 pg/mL, whereas its IC50 for cytotoxicity to BHK-21 cells was 459.45 pg/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 Gin106 and Arg133 of DENV2 NS2B/NS3 protease directly interacted with policresulen via hydrogen bonding. Conclusion： Policresulen is a potent inhibitor of DENY2 NS2B/NS3 protease that inhibits DENV2 replication in BHK-21 cells. The bind- ing mode of the protease and policresulen provides useful hints for designing new type of inhibitors against the protease.
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-DENY drugs. Here, we report a novel inhibitor of DENY2 NS2B/NS3 protease and its antiviral action. Methods： An enzymatic inhibition assay was used for screening DENY2 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 （NIO00 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 ICso of 0.48 pg/mL. Furthermore, policresulen inhibited DENV2 replication in BHK-21 cells with ICso of 4,99 pg/mL, whereas its IC50 for cytotoxicity to BHK-21 cells was 459.45 pg/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 Gin106 and Arg133 of DENV2 NS2B/NS3 protease directly interacted with policresulen via hydrogen bonding. Conclusion： Policresulen is a potent inhibitor of DENY2 NS2B/NS3 protease that inhibits DENV2 replication in BHK-21 cells. The bind- ing mode of the protease and policresulen provides useful hints for designing new type of inhibitors against the protease.