Develop Effective Inhibitors of Anthrax Lethal Factor

描述（由申请人提供）：本项目的目的是开发，加快时间表，这些炭疽致死因子（LF）的抑制剂，这是有效和安全的人类使用。美国政府已经强有力地表示，有效治疗炭疽是国家的一项关键优先事项。作为回应，PI与主要合作者密切合作，开发了一个逻辑性强、密切相关的计划，该计划将导致成熟的药物样化合物。该模块化程序采用最新和最先进的结构，酶学，NMR和计算机方法。开发将通过基本的、无缝交互的模块进行，互补功能之间具有定义良好的接口。整体模块包括：通过虚拟配体筛选来筛选已知的市售化学结构（柔性LF蛋白-配体对接）以鉴定有限数量的潜在抑制剂，在酶学和NMR研究中测试所选结构的效力，通过体外和细胞测试选择具有最小细胞毒性和对人基质金属蛋白酶（MMP）的影响的抑制剂，在埃水平上对所述聚烯烃-LF复合物进行共结晶和结构分析，并随后对化学结构进行细化以实现最佳拟合。在涉及上述每个模块的两到三次迭代之后，团队将重新评估先导化合物，并在将候选药物转入临床试验之前执行所需的基准。我们的五个具体目标是：（I）建立纯化的人MMPs的综合组;（II）检查LF的底物偏好并鉴定其潜在肽底物和蛋白质靶标的结构;（III）设计能够有效阻断LF的切割功能的最有效和选择性的抑制剂;（IV）测试目标III中选择的LF抑制剂的衍生文库的效力;（V）检测优化的、微调的化合物的细胞毒性和抑制效力，并鉴定LF的药物样先导抑制剂以用于继续的药物精制和临床试验。我们相信，由此产生的LF拮抗剂将有效和安全地提供如此迫切需要的治疗。

DESCRIPTION (provided by applicant): The purpose of this project is to develop, on an expedited schedule, those inhibitors of anthrax lethal factor (LF) which are both effective and safe for use by humans. The US government has forcibly stated that an effective treatment for anthrax is a key national priority. In response, the PI, in close coordination with the key collaborators, has developed a logical, closely interrelated program, which will lead to mature, drug-like compounds. This modular program employs the most current and advanced structural, enzymological, NMR and in silico methodologies. The development will be conducted through essential, seamlessly interacting modules with well-defined interfaces between complementary functions. The integral modules are: sieving the known, commercially available chemical structures by virtual ligand screening (flexible LF protein-ligand docking) to identify the limited number of potential inhibitors, testing the potency of the selected structures in enzymological and NMR studies, selecting via the in vitro and cell tests the inhibitors with minimal cytotoxicity and effect on human matrix metalloproteinases (MMPs), performing co-crystallization and structural analyses at the angstrom level of the inhibitor-LF complexes and the subsequent refinement of the chemical structures to accomplish the best fit. Following two or three iterations involving each of the above modules, the team will re-evaluate the lead compound(s) and perform the required benchmarks prior to moving the drug candidate(s) into clinical trials. Our five specific aims are: (I) establish a comprehensive panel of purified human MMPs; (II) examine the substrate preferences of LF and identify the structure of its potential peptide substrates and protein targets; (III) design the most potent and selective inhibitors capable of efficiently blocking the cleavage function of LF; (IV) test the potency of the derivative library of LF inhibitors selected in Aim III; (V) examine cytotoxicity and inhibitory potency of optimized, fine-tuned compounds, and to identify the drug-like lead inhibitor(s) of LF for continued drug refinement and clinical trials. We are confident that the resulting LF antagonist(s) will efficiently and safely provide the therapy that is so urgently required.