Develop Effective Inhibitors of Anthrax Lethal Factor

描述(申请人提供)：该项目的目的是加速开发炭疽致死因子(LF)的抑制剂，这些抑制剂对人类来说既有效又安全。美国政府强行表示，有效治疗炭疽病是国家的关键优先事项。作为回应，PI与主要合作者密切协调，制定了一个合乎逻辑、密切相关的计划，这将导致成熟的、类似药物的化合物。这一模块化计划采用了最新和最先进的结构、酶、核磁共振和硅胶方法。开发将通过基本的、无缝交互的模块进行，这些模块在补充功能之间具有明确定义的接口。集成模块包括：通过虚拟配体筛选(灵活的LF蛋白质-配体对接)筛选已知的商业可用化学结构，以确定有限数量的潜在抑制剂；测试所选结构在酶学和核磁共振研究中的效力；通过体外和细胞测试选择细胞毒性和对人类基质金属蛋白酶(MMPs)影响最小的抑制剂；在Angstrom水平上进行抑制剂-LF复合体的共结晶和结构分析，并随后对化学结构进行优化，以实现最佳匹配。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(S)，并在将候选药物(S)转移到临床试验之前执行所需的基准。我们的五个具体目标是：(I)建立一个全面的纯化人MMPs专家组；(Ii)检测LF的底物偏好并确定其潜在的肽底物和蛋白质靶点的结构；(Iii)设计能够有效阻断LF切割功能的最有效和选择性最高的抑制剂；(Iv)测试Aim III中选定的LF抑制剂衍生库的效力；(V)检测优化、微调化合物的细胞毒性和抑制效力，并鉴定LF的类药物先导抑制剂(S)，以便继续进行药物精制和临床试验。我们相信，由此产生的LF拮抗剂(S)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.