Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
Bioorg Med Chem Lett. 2011 Jul 1;21(13):3943-6. doi: 10.1016/j.bmcl.2011.05.021. Epub 2011 May 14.
Mycobacterium tuberculosis pantothenate synthetase is a potential anti-tuberculosis target, and a high-throughput screening system was previously developed to identify its inhibitors. Using a similar system, we screened a small library of compounds and identified actinomycin D (ActD) as a weak inhibitor of pantothenate synthetase. A new method was established to discover more effective inhibitors by determining the molecular mechanism of ActD inhibition followed by structure-based virtual screening. The molecular interaction of inhibition was determined by circular dichroism and tryptophan fluorescence quenching. The structure-based search and virtual screening were performed using the Molecular Operating Environment (MOE) program and SYBYL 7.5, respectively. Two inhibitors were identified with an IC(50) for pantothenate synthetase that was at least ten times better than that of ActD.
分枝杆菌泛酸合酶是一个有潜力的抗结核药物靶点，我们先前开发了一种高通量筛选系统来识别其抑制剂。利用类似的系统，我们筛选了一个小型化合物文库，发现放线菌素 D（ActD）是一种较弱的泛酸合酶抑制剂。通过确定 ActD 抑制的分子机制，然后进行基于结构的虚拟筛选，我们建立了一种新的方法来发现更有效的抑制剂。通过圆二色性和色氨酸荧光猝灭来确定抑制的分子相互作用。使用分子操作环境（MOE）程序和 SYBYL 7.5 分别进行基于结构的搜索和虚拟筛选。鉴定出两种抑制剂，其对泛酸合酶的半数抑制浓度（IC50）至少比 ActD 好十倍。
