M tuberculosis Membrane Protein Pharmaceutical Targets

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，而大多数生物体基因组的25%至30%编码膜蛋白-此外，膜蛋白比水溶性蛋白更频繁地成为有效的药物靶点; 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。检测开发将能够针对小分子进行筛选，这对理解功能很重要，对结构研究也可能很重要。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。

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.