Inhibitor of FtsZ Polymerization in M. tuberculosis

结核分枝杆菌中 FtsZ 聚合的抑制剂

• 批准号: 6496584
• 负责人: Robert C Reynolds
• 金额: $ 55.12万
• 依托单位: SOUTHERN RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-15 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6496584
• 关键词: Mycobacterium tuberculosis; aminopyridines; antitubercular agents; bioassay; biological models; chemical kinetics; chemical structure function; chemical synthesis; cytotoxicity; diazepine; drug design /synthesis /production; drug screening /evaluation; electron microscopy; genetic strain; guanosinetriphosphatases; hydrolysis; laboratory mouse; macrophage; model design /development; polymerization; pteridines; tubulin

DESCRIPTION (Provided by the applicant): Development of new antitubercular agents is of critical importance worldwide. Our program has identified a new class of inhibitor of Mycobacterium tuberculosis(Mtb) that inhibits a novel protein not presently targeted by current antitubercular agents. The 2-alkoxy-carbonylamino-pyridines (2-ACPs) potently inhibit the growth of Mtb with an MIC99 (SRI-3072) as low as 0.15 microgram/ml (0.28 micromolar). Furthermore, SRI-3072 shows bactericidal activity, and shows significant activity in a murine-derived macrophage model with an EC90 & EC99 of 0.12 and 1.42 microgram/ml respectively. These analogs also show selective activity against Mtb versus a mammalian cell line. This program has successfully identified the target of these agents, the mycobacterial tubulin homolog FtsZ. The target protein has been cloned, expressed and isolated in quantities sufficient for development of in vitro polymerization and GTP hydrolysis assays. Three compounds, SRI-3072, SRI-76 14, and colchicine have been shown to inhibit polymerization of Mtb FtsZ in a dose dependent manner with IC50S of 50 uM, 60 uM, and 100 uM respectively. Furthermore. we have shown that SRI-7614 affects Mtb FtsZ polymerization by electron microscopy. SRI-7614 has also been shown to be active vs. a panel of single drug-resistant Mtb strains. We currently have crystal structures of Mtb FtsZ bound to citrate, GTPgS, and GDP. To date, about 200 2-ACP analogs have been screened in vitro against Mtb H37Rv. We have developed a SAR profile that will allow the preparation of more selective and more potent antitubercular agents. In this application, we propose to continue development of the 2-ACP class through preparations of new analogs of the more potent and selective subclasses, the 3-deaza-pteridines (priority), and the pyridodiazepines (backup). We will carefully evaluate these compounds for activity and selectivity in various in vitro assays including an in vitro Mtb H37Ra assay, an in vitro Mtb FtsZ polymerization and GTPase assay, an in vitro tubulin polymerization assay and a mammalian cell toxicity assay. Selected active agents will be further screened in an h in vitro macrophage model and a Mtb mouse model. The effect of inhibitors on FtsZ polymerization will be analyzed using electron microscopy. Data from the biological screening and the EM structural studies will feed back into compound design in an interactive, iterative drug design cycle that critically focuses on antibacterial potency and selectivity.

描述(申请人提供)：新型抗结核药物的开发 代理商在世界范围内具有至关重要的意义。我们的节目发现了一个新的 一类结核分枝杆菌(Mtb)抑制剂，抑制一种新的 目前未被当前抗结核药物靶向的蛋白质。这个 2-烷氧基甲酰氨基吡啶(2-ACPs)对结核分枝杆菌的生长有明显的抑制作用 MIC99(SRI-3072)低至0.15微克/毫升(0.28微摩尔)。 此外，SRI-3072具有杀菌活性，并显示出显著的杀菌活性 小鼠巨噬细胞模型的活性，其EC90和EC99分别为0.12和 1.42微克/毫升。这些类似物也显示出选择性活性。 对抗结核分枝杆菌与哺乳动物细胞系。这个项目已经成功地 确定了这些药物的靶标，分枝杆菌微管蛋白同系物FtsZ。 目的蛋白已被大量克隆、表达和分离 足以发展体外聚合和GTP水解 化验。三种化合物，SRI-3072，SRI-76 14和秋水仙碱已被证明 抑制Mtb FtsZ聚合呈剂量依赖性，IC50S为50 嗯，分别是60微米和100微米。更重要的是。我们已经证明了SRI-7614 电子显微镜观察对Mtb-FtsZ聚合的影响。SRI-7614也一直是 与一组单一耐药的结核分枝杆菌菌株相比，显示出活性。我们 目前具有与柠檬酸、GTPgS和GDP结合的Mtb FtsZ的晶体结构。 到目前为止，已有约200个2-ACP类似物在体外筛选出抗结核分枝杆菌H37Rv。 我们已经开发了一种SAR配置文件，将允许准备更多 选择性更强的抗结核药物。在此应用程序中，我们 建议通过筹备新的2-ACP课程继续发展 更有效和更有选择性的亚类的类似物，3-去氮衍生物 (优先)，以及吡哆二氮卓类药物(备用)。我们将仔细评估这些措施 化合物在各种体外试验中的活性和选择性，包括 体外Mtb H37Ra试验，体外Mtb FtsZ聚合和GTPase测定， 体外微管蛋白聚合试验和哺乳动物细胞毒性试验。 选定的活性物质将在体外巨噬细胞中进一步筛选 模型和Mtb小鼠模型。阻聚剂对FtsZ聚合的影响 将用电子显微镜进行分析。来自生物筛查的数据 EM结构研究将反馈到化合物设计中 交互、迭代的药物设计周期，关键是 抗菌效力和选择性。