摘要
目的发现结核分枝杆菌Pks13(MtPks13)的抑制剂,为后续针对以MtPks13为靶点开发抗结核药物奠定了基础。方法表达MtPks13-TE蛋白并优化其酶活测定方法,构建MtPks13抑制剂高通量筛选模型;与海分枝杆菌表型筛选方法联用,对5万个化合物进行筛选,将获得的抑制剂IMB-7142进行IC_(50)以及酶动力学性质的测定;表面等离子共振(surface plasmon resonance,SPR)实验以及分子对接模型研究阳性化合物IMB-7142与MtPks13-TE蛋白是否能发生相互作用及其作用位点;用菌液稀释法评价IMB-7142对结核标准株的抗结核活性。结果成功构建了MtPks13抑制剂筛选模型,并发现了一个对MtPks13有抑制作用,同时抑制海分枝杆菌生长的化合物IMB-7142,该抑制剂能与MtPks13-TE蛋白发生相互作用,具有较好的体外抗结核活性。结论成功构建了稳定的MtPks13抑制剂高通量筛选模型,并且应用该模型筛选得到的一个抑制剂同时具有抗结核活性,为后续开发以MtPks13为靶点抗结核药物提供了思路。
Objective To discover inhibitors targeting Mycobacterium tuberculosis Pks13,and provide a foundation for the development of novel anti-tuberculosis drugs.Methods Mycobacterium tuberculosis Pks13-TE protein was overexpressed,and then the purified MtPks13-TE was used to establish a high-throughput screening model.In combination with the phenotype screening method of marine Mycobacterium,50,000 compounds were screened,and the IC_(50) and enzyme kinetic properties of the lead candidate inhibitor,IMB-7142,were determined.Surface plasmon resonance(SPR)experiment and molecular docking model were used to study whether the positive compound IMB-7142 could interact with MtPks13-TE protein and,if so,its interaction sites.The anti-tuberculosis activity was evaluated by the broth dilution method.Results The screening model for MtPks13 inhibitors was constructed,and the compound IMB-7142 was found to inhibit MtPks13 and the growth of marine Mycobacterium.IMB-7142 could interact with MtPks13-TE and had anti-tuberculosis activity.Conclusion A stable highthroughput screening model for MtPks13 inhibitors was constructed,and the inhibitor obtained using this screening model exhibited anti-tuberculosis activity,which provided a starting point for the subsequent development of antituberculosis drugs targeting MtPks13.
作者
王潇
蒙建州
关艳
刘思含
张佳炜
李晓辉
刘忆霜
Wang Xiao;Meng Jian-zhou;Guan Yan;Liu Si-han;Zhang Jia-wei;Li Xiao-hui;Liu Yi-shuang(National Laboratory for Screening New Microbial Drugs,Institute of Medicinal Biotechnology,Peking Union Medical College and Chinese Academy of Medical Sciences,Beijing 100050;Shanghai Key Laboratory of Tuberculosis,Shanghai Pulmonary Hospital,Tongji University School of Medicine,Shanghai 200433;School of Basic Medical Science,Jiamusi University,Jiamusi 154007)
出处
《中国抗生素杂志》
CAS
CSCD
北大核心
2022年第10期1038-1044,共7页
Chinese Journal of Antibiotics
