Project 1 UIC Targeting Protein Degradation ClpC1 ATPase

项目摘要/摘要 项目1的中心目标是确定候选ClpC1调节剂，以转化为临床抗肿瘤药物。 结核病(TB)药物。正如NIAID主任最近强调的那样，迫切需要新的口服结核病药物。 我们和其他人的工作已经确定ClpC1是治疗结核病的一个新的蛋白质靶点。已知天然产物 (NP)ClpC1调节剂在体外表现出良好的抗结核分枝杆菌(Mtb)特性，提示 作用于这一目标的结核病药物将对多药耐药/广泛耐药(MDR/XDR)有效- 结核病，有可能缩短治疗时间。已知的ClpC1调节剂的药代动力学(PK)特性， 优胜霉素、红霉素、环素A和拉霉素A排除了将这些NP环肽直接用于口服结核病的可能性 毒品。因此，项目1试图确定和开发具有潜在治疗作用的新型口服生物可用ClpC1调节剂 在本《经济、社会和文化权利国际公约》设想的结核病治疗方案内使用。识别潜在客户的不同方法包括： 发现新的NPs和新的小分子ClpC1调节剂，并优化已知NPs的性质。每个人 该方法将利用多肽化学、基于结构的药物设计(SBDD)、NP 发现和基于片段的药物设计。Lead发现和优化将利用四个正交生物物理 了解与ClpC1结合的关键因素的方法：表面等离子激元共振(SPR)、X射线 结晶学、核磁共振和低温电子显微镜。纳米胶囊化工作旨在优化优生素的暴露，以充分 探索其在结核分枝杆菌感染小鼠中的疗效。UIC项目1团队处于独特的地位，可以遵循这些 目标：开创新的NP技术，在药物发现和领导验证中具有独特的实用价值 导致了从放线菌中发现了优生素和红霉素。UIC还进行了广泛的工作，以获得 证明了Mtb ClpC1蛋白酶的脆弱性的机械论见解，支持了建议的 活动。UIC进行的研究将有助于推动ClpC1作为抗结核药物靶点的翻译。这个 项目1的具体目标在《经济、社会和文化权利国际公约》内也有很强的协作关系。[目标1]的目的是优化 NP ClpC1调节剂以探讨其抗结核杆菌的效果。这涉及与Myongji(扩大规模)的合作 生产)和普林斯顿大学(增强体内药效的纳米胶囊)。纯净度和NP完整性 寡肽的分析将采用UIC的q核磁共振方法。将进行PK和疗效研究 W/内核A/B。[目标2]将采用不同的方法(大规模NP分离和表征；NP启发 结构/合成孔径雷达引导设计，核磁共振筛选)，以确定新的和口服可用的ClpC1调节剂。该阵列 方法包括UIC的分子网络，通过与礼来公司合作进行基于片段的筛查，ClpC1 功能分析与项目2，核心A，肽和药物化学，SBDD，体外ADME，和 鼠标主键。[目的3]探讨影响NPs、人工合成环肽与ClpC1亲和力的关键因素 通过SPR、核磁共振、低温电子显微镜和共晶X射线分析等手段对小分子进行了分析。AIM 3也将优化 从AIM 2中产生的线索，然后是候选人选择和通过核心C的IND使能研究。

Project Summary/Abstract The central objective of Project 1 is to identify candidate ClpC1 modulators for translation into clinical anti- tuberculosis (TB) drugs. As recently emphasized by the NIAID Director, new oral TB drugs are sorely needed. Our work and that of others has identified ClpC1 as a novel protein target for TB treatment. Known natural product (NP) ClpC1 modulators display favorable in vitro anti-Mycobacterium tuberculosis (Mtb) properties, suggesting that TB drugs acting on this target would have utility against multi-drug/extensively drug-resistant (MDR/XDR)- TB, with potential to shorten treatment. The pharmacokinetic (PK) properties of the known ClpC1 modulators, ecumicin, rufomycin, cyclomarin-A and lassomycin, preclude direct use of these NP cyclic peptides as oral TB drugs. Thus, Project 1 seeks to identify and develop novel orally bioavailable ClpC1 modulators with potential for use within the TB treatment regimens envisioned in this CETR. Diverse approaches to lead identification include: identifying new NPs and novel small molecule ClpC1 modulators, and optimizing properties of known NPs. Each approach will leverage leading edge technologies in peptide chemistry, structure-based drug design (SBDD), NP discovery, and fragment-based drug design. Lead finding and optimization will utilize four orthogonal biophysical approaches to understand key factors in binding to ClpC1: surface plasmon resonance (SPR), X-ray crystallography, NMR, and Cryo-EM. Nanoencapsulation work aims at optimizing exposure of ecumicin, to fully explore its efficacy profile in Mtb-infected mice. The UIC Project 1 team is uniquely positioned to follow these objectives, having pioneered new NP technologies with unique utility in drug discovery and lead validation that led to the discovery, from actinomycetes, of ecumicin and rufomycin. UIC also conducted extensive work to gain mechanistic insights that demonstrate the vulnerability of Mtb ClpC1 protease, underpinning the proposed activities. Research conducted at UIC will help drive the translation of ClpC1 as an anti-Mtb drug target. The Specific Aims of Project 1also have strong collaborative ties within the CETR. [AIM 1] is to optimize exposure of NP ClpC1 modulators to explore their anti-Mtb efficacy. This involves collaborations with Myongji (scale-up production) and Princeton University (nano-encapsulation for enhanced in vivo efficacy). Purity and NP integrity analysis of the oligopeptides will employ UIC’s qNMR methodology. PK and efficacy studies will be conducted w/Cores A+B. [AIM 2] will employ diverse approaches (large-scale NP isolation and characterization; NP-inspired structure-/SAR-guided design, NMR screening) to identify novel and orally available ClpC1 modulators. The array of methods involves Molecular Networking at UIC, fragment-based screening via collaboration with Eli Lilly, ClpC1 functional assays with Project 2, and at Core A, peptide and medicinal chemistry, SBDD, in vitro ADME, and mouse PK. [AIM 3] seeks to evaluate key factors in ClpC1 binding affinity of NPs, synthetic cyclic peptides and small molecules by using SPR, NMR, Cryo-EM, and co-crystallization X-ray analysis. Aim 3 will also optimize leads emerging from Aim 2, followed by candidate selection and IND-enabling studies through Core C.