Screen for Mycobacterium Tuberculosis (RMI)

描述(申请人提供)：据估计，世界上三分之一的人口感染了结核病，每年约有800万新病例。耐多药结核病的定义是对两种或两种以上一线抗结核药物耐药的形式，已成为一个日益严重的健康问题，特别是在感染艾滋病毒的人中。这些形式的结核病往往是致命的，也更难治疗。因此，针对新途径的分子研究工具或药物的开发变得越来越重要。泛酸的生物合成途径就是这样一个靶点。泛酸是分枝杆菌合成辅酶A和酰基载体蛋白的重要前体，而辅酶A和酰基载体蛋白参与脂肪酸合成。泛酸合成酶由panc基因编码，催化细菌中泛酸的形成。研究了结核分枝杆菌泛酸合成酶的酶学性质，并确定了其高分辨晶体结构。最近，人们发现了几种抑制这种酶的化合物。不幸的是，这些化合物都不影响结核分枝杆菌的体外生长。为了满足分子研究工具研究结核分枝杆菌泛酸途径的需要，本提案的具体目的是：1)将现有的泛酸合成酶HTS检测转移到分子文库筛选中心网络。。2)在TAACF提供的一系列验证性试验中跟踪HTS试验的结果，以评估它们在体外和体内对结核分枝杆菌的疗效。3)研究这些新型抑制剂对耻垢分枝杆菌泛酸途径的影响。4)将这些化合物与泛酸合成酶共结晶，研究它们的结合方式。

DESCRIPTION (provided by applicant): It is estimated that one third of the world's population is infected with tuberculosis (TB), with about 8 million new cases annually. Multi-drug resistant TB, defined as forms resistant to two or more of the front line anti-TB agents, has become an increasing health problem, especially in HIV infected individuals. These forms of TB are more often fatal and more difficult to treat. Consequently, development of molecular research tools or drugs targeting novel pathways has become increasingly important. The biosynthetic pathway of pantothenate is such a target. Pantothenate is an essential precursor in mycobacteria for the synthesis of co-enzyme A and acyl carrier protein which are involved in fatty acid synthesis. Pantothenate synthetase, encoded by the panC gene, catalyzes the formation of pantothenate in bacteria. M. tuberculosis pantothenate synthetase has been characterized enzymatically and a high resolution crystal structure has also been solved. More recently several compounds have been identified which inhibit this enzyme. Unfortunately, none of these compounds affect the growth of Mycobacterium tuberculosis in vitro. To address the need for molecular research tools to study the pantothenate pathway in M. tuberculosis the specific aims of this proposal are: 1) To transfer the existing HTS assay for pantothenate synthetase to the Molecular Libraries Screening Center Network. . 2) To follow up hits from the HTS assay in a series of confirmatory assays available through the TAACF that would evaluate their in vitro and in vivo efficacy against M. tuberculosis. 3) To study the effect of these novel inhibitors on the pantothenate pathway in M. smegmatis. 4) To co-crystallize these compounds with pantothenate synthetase to study their mode of binding