以布氏锥虫亮氨酰tRNA合成酶为靶标的抗锥虫药物筛选系统的建立

中国生物工程杂志

2009, Vol. 29

Issue (12): 13-17

技术与方法

以布氏锥虫亮氨酰tRNA合成酶为靶标的抗锥虫药物筛选系统的建立

杲光伟,姚璎,丁大中,叶龙,周虎臣,李大伟^*

上海交通大学药学院化学生物学研究所上海 200240

The Establishment of an Anti-Trypanosoma Drug Screening System with Leucyl-tRNA Synthetase as an Inhibition Target

GAO Guang-wei,YAO Ying,DING Da-zhong,YE Long,ZHOU Hu-chen,LI Da-wei

The Institute of Chemical Biology, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China

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摘要：

锥虫是人的血液寄生虫，对热带乃至拉丁美洲贫困地区影响极大，但目前的传统治疗药物存在副作用大、有效性不断降低的问题。根据亮氨酰tRNA合成酶在微生物中可作为药物靶点的事实，在新型抗锥虫药物筛选中，通过对锥虫的亮氨酰tRNA合成酶的克隆、表达和纯化，以及该酶活性测定的优化，建立了该酶抑制物的筛选系统。筛选结果表明，这一以锥虫亮氨酰tRNA合成酶为靶标的抗锥虫药物筛选系统可以有效筛选抗锥虫化合物，选出的化合物有一定的抗锥虫特异性，并可以用于化合物的进一步优化和测定其半抑制浓度。使用这一系统筛选到了对锥虫有较好抑制作用，但对人类细胞毒性较小的一系列新型化合物，因而锥虫亮氨酰tRNA合成酶很可能成为开发有效抗锥虫药物的新靶标。

关键词： 布氏锥虫; 亮氨酰tRNA合成酶; 抑制剂筛选

Abstract:

Trypanosoma is a human parasite severely affecting poor tropical areas. However, current frontline drugs for Trypanosoma treatment have severe side-effects with decreased effectiveness. Based on the fact that aminoacyl-tRNA synthetase is a bona fide drug target for several microorganisms, including bacteria and fungi, it is plausible that it may also be effective target of Trypanosoma. In this report, the Trypanosoma brucei Leucyl-tRNA synthetase (tbLeuRS) was cloned , expressed and purified to develop an in vitro enzymatic assay system. The assay conditions were further optimized for the effective screening of tbLeuRS inhibitors thus establishing an anti-Trypanosoma drug screening system targeting tbLeuRS. The results indicated that this system can be employed for the effective screening of anti-Trypanosoma drugs with satisfactory specificity. In addition, this system can also be used for compound optimization, as well as IC50 testing. Using this system a series of compounds are identified that are effective Trypanosoma inhibitors without toxicity to human cells. Therefore, targeting tbLeuRS may represent a new venue for the development of anti-Trypanosoma drugs.

Key words: Trypanosoma brucei LeuRS Screening

收稿日期: 2009-07-09 出版日期: 2009-12-21

ZTFLH:

Q819

通讯作者: 李大伟 E-mail: daweili@sjtu.edu.cn

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引用本文:

杲光伟姚璎丁大中叶龙周虎臣李大伟. 以布氏锥虫亮氨酰tRNA合成酶为靶标的抗锥虫药物筛选系统的建立[J]. 中国生物工程杂志, 2009, 29(12): 13-17.

GAO Guang-Wei, TAO Ying, DING Da-Zhong, XIE Long, ZHOU Hu-Chen, LI Da-Wei. The Establishment of an Anti-Trypanosoma Drug Screening System with Leucyl-tRNA Synthetase as an Inhibition Target. China Biotechnology, 2009, 29(12): 13-17.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2009/V29/I12/13

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