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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2013, Vol. 33 Issue (12): 51-56    
技术与方法     
以MERS-CoV主蛋白酶为靶点的药物筛选体系的建立及应用
张宁1,3, 潘丽1,3, 牛国君3, 王文明4, 周红刚2, 杨诚2,3
1. 天津科技大学生物工程学院 天津 300457;
2. 南开大学药学院 天津 300457;
3. 天津市国际生物医药联合研究院有限公司 天津 300457;
4. 南开大学生命科学学院 天津 300457
Establishment and Application of a System for Drug Screening Targeting MERS-CoV Main Proteinase
ZHANG Ning1,3, PAN Li1,3, NIU Guo-jun3, WANG Wen-ming4, ZHOU Hong-gang2, YANG Cheng2,3
1. College of Biological Engineering, Tianjin University of Science & Technology, Tianjin 300457, China;
2. College of Pharmacy, Nankai University, Tianjin 300457, China;
3. Tianjin International Joint Academy of Biomedicine Co.Ltd., Tianjin 300457, China;
4. College of Life Sciences, Nankai University, Tianjin 300457, China
 全文: PDF(766 KB)   HTML
摘要： 目的：以新型冠状病毒（MERS-CoV）主蛋白酶NSP5为靶点，应用荧光共振能量转移法（FRET）建立药物筛选体系，并进行抑制剂的筛选。方法：采用原核表达系统、利用基因重组以及蛋白表达纯化技术得到目的蛋白，FRET法检测蛋白酶活性，优化反应条件以及确定酶、底物的浓度，建立药物筛选体系，对700种化合物进行了检测。结果：应用建立的药物筛选体系，筛选到抑制率较高的化合物共6种，并测得IC50值，其中MDCCCL002330的IC50值最低，为（0.43±0.03）μmol/L。结论：建立的药物筛选体系较为理想，适用于NSP5抑制剂的筛选，促进先导化合物的研发。
关键词： NSP5;  抑制剂;  高通量筛选;  MERS-CoV    
Abstract: Objective: Targeting MERS-CoV main proteinase NSP5, a new system for drug screening was established for inhibitors screening by using fluorescence resonance energy transfer (FRET). Methods: Target protein was obtained using prokaryotic expression system and technologies of gene recombination, protein-expression and purification. Proteinase activity of target protein was examined by FRET assays. Finally, a system for drug screening was established and optimized before abundant compounds screening. Results: Using the drug screening system, eight compounds with high inhibition ratio were selected. Notably, the IC50 of MDCCCL002330 is minimal, namely 0.43μmol/L. Conclusion: The established system targeting MERS-CoV main proteinase NSP5 is suitable for inhibitors screening, which can be used to promote research and development of lead compounds.
Key words: NSP5    Inhibitors    High Throughput Screening    MERS-CoV
收稿日期: 2013-09-17 出版日期: 2013-12-25
ZTFLH:  Q819  
基金资助: 蛋白类生物药和疫苗发展专项—建立天津生物技术药物研发开放实验室和GMP中试服务平台资助项目
通讯作者: 杨诚，E-mail：cheng.yang@htmdc.org     E-mail: cheng.yang@htmdc.org
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引用本文:

张宁, 潘丽, 牛国君, 王文明, 周红刚, 杨诚. 以MERS-CoV主蛋白酶为靶点的药物筛选体系的建立及应用[J]. 中国生物工程杂志, 2013, 33(12): 51-56.

ZHANG Ning, PAN Li, NIU Guo-jun, WANG Wen-ming, ZHOU Hong-gang, YANG Cheng. Establishment and Application of a System for Drug Screening Targeting MERS-CoV Main Proteinase. China Biotechnology, 2013, 33(12): 51-56.

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https://manu60.magtech.com.cn/biotech/CN/    或    https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I12/51

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