结核分枝杆菌β-内酰胺酶BlaC抑制剂筛选模型的建立及应用

doi:10.5246/jcps.2016.03.022

中国药学(英文版) ›› 2016, Vol. 25 ›› Issue (3): 189-195.DOI: 10.5246/jcps.2016.03.022

结核分枝杆菌β-内酰胺酶BlaC抑制剂筛选模型的建立及应用

刘忆霜¹, 郑佳音², 黄树超¹, 关艳¹, 肖春玲^1*

1. 中国医学科学院北京协和医学院医药生物技术研究所, 国家新药(微生物)筛选实验室, 北京 100050
2. 首都医科大学, 神经生物学系, 北京 100069

收稿日期:2016-01-18 修回日期:2016-02-17 出版日期:2016-03-29 发布日期:2016-02-24
通讯作者: Tel.: 861063020226, E-mail: xiaocl318@163.com
基金资助:
Fundamental Research Funds for Central Public Welfare Research Institutes (Grant No. 2015PT350001) and National Major Scientific and Technological Special Project for “Significant New Drugs Development” (Grant No. 2015ZX09102007-009).

Establishment and application of the screening model of the Mycobacterium tuberculosis β-lactamase BlaC inhibitors

Yishuang Liu¹, Jiayin Zheng², Shuchao Huang¹, Yan Guan¹, Chunling Xiao^1*

1. National Key Laboratory for Screening New Microbial Drugs, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China
2. Department of Neurobiology, Capital Medical University, Beijing 100069, China

Received:2016-01-18 Revised:2016-02-17 Online:2016-03-29 Published:2016-02-24
Contact: Tel.: 861063020226, E-mail: xiaocl318@163.com
Supported by:
Fundamental Research Funds for Central Public Welfare Research Institutes (Grant No. 2015PT350001) and National Major Scientific and Technological Special Project for “Significant New Drugs Development” (Grant No. 2015ZX09102007-009).

摘要/Abstract

摘要：

随着多药耐药和广泛耐药结核的迅速蔓延,迫切需要寻求对抗结核的新手段。结核分枝杆菌对β-内酰胺类抗菌药物的固有耐药性限制了β-内酰胺类抗菌药物在结核治疗上的应用, 而A类(Ambler分类)β-内酰胺酶BlaC的表达是结核分枝杆菌对β-内酰胺类抗菌药物耐药的最主要原因。抑制BlaC可以恢复结核分枝杆菌对于β-内酰胺类抗菌药物的敏感性, 因此BlaC抑制剂可能成为新型的抗结核药物增效剂, 并成为治疗结核的新途径。在本研究中, 表达并纯化了结核分枝杆菌BlaC, 并以此为基础构建了BlaC抑制剂的筛选模型, 通过条件优化使该模型适用于高通量筛选。通过对26 400个化合物样品的筛选, 得到了22个BlaC抑制剂, 阳性率为0.083%。这一研究为可用于临床的新型抗结核药物增效剂的发现奠定了基础。

关键词: 结核分枝杆菌, β-内酰胺酶, BlaC, 高通量筛选模型, 抗结核药物增效剂

Abstract:

With the continuous emergence and rapid spread of multidrug-resistant and extensively-drug-resistant Mycobacterium tuberculosis strains, it is imperative to develop novel therapies against this bacterium. The intrinsic β-lactam resistance ofM. tuberculosis is primarily due to the production of an Ambler class-A β-lactamase BlaC, which limits the application of β-lactam antibiotics in the treatment of tuberculosis. Therefore, the inhibitors of BlaC could be novel anti-tuberculosis drug synergistic agents to recover the sensibility of M. Tuberculosis to the β-lactam antibiotics. In the present study, BlaC of M. tuberculosis was expressed and purified to establish a screening model of the BlaC inhibitors. The screening conditions were determined, and the screening model was evaluated to fit for the high throughput screening. A total of 22 BlaC inhibitors were screened out from 26 400 compound samples with a positive rate of 0.083%. Taken together, our findings lay the foundation for the discovery of novel anti-tuberculosis drug synergistic agents in clinic.

Key words: Mycobacterium tuberculosis, β-Lactamase, BlaC, High-through screening model, Anti-tuberculosis drug synergistic agents

中图分类号:

R965

Supporting:

刘忆霜, 郑佳音, 黄树超, 关艳, 肖春玲. 结核分枝杆菌β-内酰胺酶BlaC抑制剂筛选模型的建立及应用[J]. 中国药学(英文版), 2016, 25(3): 189-195.

Yishuang Liu, Jiayin Zheng, Shuchao Huang, Yan Guan, Chunling Xiao. Establishment and application of the screening model of the Mycobacterium tuberculosis β-lactamase BlaC inhibitors[J]. Journal of Chinese Pharmaceutical Sciences, 2016, 25(3): 189-195.

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结核分枝杆菌β-内酰胺酶BlaC抑制剂筛选模型的建立及应用

Establishment and application of the screening model of the Mycobacterium tuberculosis β-lactamase BlaC inhibitors

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