M tuberculosis Membrane Protein Pharmaceutical Targets

结核分枝杆菌膜蛋白药物靶点

• 批准号: 7561796
• 负责人: TIMOTHY A CROSS
• 金额: $ 176.58万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-20 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7561796
• 关键词: tuberculosis; Membrane; Protein; Pharmaceutical; Targets

DESCRIPTION (provided by applicant): This Proposal is designed to address: 1) an important disease that needs novel drugs - no new drugs for Mycobacterium tuberculosis in 40 years and multi-drug resistant strains as well as extreme drug resistant strains are becoming more common; 2) the lack of structural information for an entire class of drug targets, the membrane proteins - less than 1% of known protein structures are membrane proteins, while 25 to 30% of the genome of most organisms code for membrane proteins - in addition, membrane proteins are more frequently effective drug targets than water-soluble proteins; 3) biological, functional and structural characterization of validated targets - we will characterize only those proteins that are essential for Mtb growth and by targeting membrane proteins, especially the outer membrane proteins, access to the drug targets will not require transport across the bacterial membranes. 4) a gap in screening technology - new small molecule screening technologies based on solution and solid state NMR spectroscopy will be developed specifically for membrane proteins. We have developed an Initial Target List from preliminary results and from literature on essential genes, virulence factors, identification of outer membrane proteins and numerous individual studies on specific potential targets. Some of these proteins are already validated as high potential pharmaceutical targets, these form a Prioritized Target List that will allow all of the Projects and Cores to initiate their efforts on the first day of funding. From biological function (Project 1) to assay development (Project 2) to structural characterization (Project 3) these activities will work closely together. Assays coupled with molecular structure will help establish structure-activity-relationships. Assay development will enable screening against small molecules important for understanding function and potentially important for structural studies. The assays we develop and the ligands we identify will fuel biological experiments designed to understand the life and infection cycle of Mtb. These ligands will be useful as lead compounds in drug discovery, and while this is beyond the scope of this Program, this team will protect the intellectual property for those who may want to pursue the development of drugs for these membrane protein targets. To accomplish these goals a unique team of investigators has been brought together with extensive knowledge of: Mycobacterium tuberculosis, essential Mtb genes, membrane protein physiology, molecular biology, biophysics, and structural characterization. The Program offers access to two of the premier NMR facilities in the world along with their expertise in methodology and technology development, in addition, the Team brings with it unique expression libraries of Mtb membrane proteins and the superb screening facilities and expertise of the Burnham Institute. PROJECT 1: TARGET IDENTIFICATION (NIEDERWEIS, M) PROJECT 1 DESCRIPTION (provided by applicant): More than 60% of all drug targets are membrane proteins. However, only one of the current antituberculous drugs, ethambutol, targets membrane proteins of M. tuberculosis, indicating that this vast potential is largely untapped for M. tuberculosis. The recent discovery of a new diarylquinoline drug that targets the ATP synthase of M. tuberculosis and exceeds the activities of the two main TB drugs isoniazid and rifampin provides proof of principle that this assumption is correct. Therefore, we have selected 11 out of approximately 800 putative inner membrane proteins of M. tuberculosis, which are essential for growth of M. tuberculosis in vitro or in mice. These proteins were selected to cover a broad variety of functions, such as transport of essential nutrients, cell division, export of proteins and cell wall and lipid biosynthesis. Furthermore, they are sufficiently small to be amenable to current solid-state NMR methods. Outer membrane proteins offer the tremendous advantage as drug targets that inhibitors may not need to cross the outer membrane which is an extremely efficient permeability barrier in mycobacteria. In addition, they are likely to represent novel drug targets because they do not appear to show any similarity to other proteins. Therefore, we included seven putative outer membrane proteins in our list of initial targets, six of which have no known function. The goal of this project is to validate the potential of the 18 selected membrane proteins as drug targets, to elucidate the physiological function of the six proteins with no known homologs and to identify putative interaction partners for some of these proteins.

描述（申请人提供）：本建议书旨在解决：1）需要新药的重要疾病-结核分枝杆菌40年无新药，多重耐药菌株和极端耐药菌株越来越常见; 2）缺乏整个一类药物靶标的结构信息，膜蛋白-不到1%的已知蛋白质结构是膜蛋白，而大多数生物体的基因组的25%至30%编码膜蛋白-此外，膜蛋白是比水溶性蛋白更常见的有效药物靶标; 3）经验证的靶标的生物学、功能和结构表征-我们将仅表征对于Mtb生长必需的那些蛋白质，并且通过靶向膜蛋白，特别是外膜蛋白，到达药物靶点将不需要穿过细菌膜的运输。4)筛选技术方面的一个空白-将专门针对膜蛋白开发基于溶液和固态NMR光谱的新的小分子筛选技术。我们根据初步结果和关于必需基因、毒力因子、外膜蛋白鉴定以及关于特定潜在靶点的众多单独研究的文献，制定了一份初始靶点清单。这些蛋白质中的一些已经被验证为高潜力的药物靶点，这些形成了一个优先目标列表，将允许所有项目和核心在资助的第一天就开始工作。从生物学功能（项目1）到检测开发（项目2）再到结构表征（项目3），这些活动将密切合作。结合分子结构的分析将有助于建立结构-活性-关系。检测开发将能够筛选对理解功能重要的小分子，并可能对结构研究重要。我们开发的检测方法和我们鉴定的配体将为旨在了解结核分枝杆菌生命和感染周期的生物实验提供动力。这些配体将作为药物发现中的先导化合物，虽然这超出了本计划的范围，但该团队将保护那些可能希望开发这些膜蛋白靶点药物的人的知识产权。为了实现这些目标，一个独特的研究团队聚集在一起，拥有广泛的知识：结核分枝杆菌，必要的Mtb基因，膜蛋白生理学，分子生物学，生物物理学和结构表征。该计划提供了访问两个首屈一指的核磁共振设施在世界上沿着与他们的专业知识在方法和技术开发，此外，该小组带来了独特的表达文库结核分枝杆菌膜蛋白和一流的筛选设施和专业知识的伯纳姆研究所。 项目1：目标识别（尼德尔维斯，密歇根州） 项目1描述（由申请人提供）：超过60%的药物靶标是膜蛋白。然而，目前的抗结核药物中只有乙胺丁醇靶向于结核分枝杆菌膜蛋白。结核病，表明这一巨大的潜力在很大程度上是未开发的M。结核最近发现的一种新的二芳基喹啉类药物靶向的ATP合酶的M。结核病和超过活动的两个主要结核病药物异烟肼和利福平提供了原则证明，这一假设是正确的。因此，我们从大约800种推定的M内膜蛋白中选择了11种。结核分枝杆菌生长所必需的蛋白质。在体外或在小鼠中的结核病。选择这些蛋白质以涵盖广泛的功能，例如必需营养素的运输、细胞分裂、蛋白质和细胞壁的输出以及脂质生物合成。此外，它们足够小，可以适用于当前的固态NMR方法。外膜蛋白作为药物靶点提供了巨大的优势，抑制剂可能不需要穿过外膜，外膜是分枝杆菌中极其有效的渗透屏障。此外，它们可能代表新的药物靶点，因为它们似乎与其他蛋白质没有任何相似性。因此，我们在我们的初始靶点列表中包括了七种假定的外膜蛋白，其中六种没有已知的功能。该项目的目标是验证18个选定的膜蛋白作为药物靶点的潜力，阐明没有已知同源物的六种蛋白质的生理功能，并确定其中一些蛋白质的假定相互作用伴侣。