Project 1 UIC Targeting Protein Degradation ClpC1 ATPase

项目 1 UIC 靶向蛋白质降解 ClpC1 ATPase

• 批准号: 10595582
• 负责人: Scott G. Franzblau
• 金额: $ 44.53万
• 依托单位: GLOBAL ALLIANCE FOR TB DRUG DEVELOPMENT
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10595582
• 关键词: ATP phosphohydrolase; Actinobacteria class; Affinity; Antitubercular Agents; Binding; Biological Assay; Biological Availability; Chemistry; Clinical; Collaborations; Consultations; Cryoelectron Microscopy; Crystallization; Cyclic Peptides; Data; Drug Design; Drug Kinetics; Drug Targeting; Drug resistant Mycobacteria Tuberculosis; Encapsulated; Ensure; Evaluation; Extreme drug resistant tuberculosis; Fermentation; Genetic Transcription; Goals; In Vitro; Lead; Leadership; Libraries; Methodology; Methods; Molecular; Mus; Mycobacterium tuberculosis; National Institute of Allergy and Infectious Disease; Natural Products; Oligopeptides; Oral; Oral Tuberculosis; Peptide Hydrolases; Peptides; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Positioning Attribute; Production; Property; Proteins; Regimen; Research; Ribosomes; Roentgen Rays; Role; Safety; Series; Sterilization; Structure; Structure-Activity Relationship; Surface Plasmon Resonance; Technology; Translations; Treatment Protocols; Tuberculosis; Universities; Validation; Vertebral column; Work; X-Ray Crystallography; analog; biophysical techniques; candidate identification; candidate selection; data integration; design; drug action; drug candidate; drug discovery; drug-sensitive; efficacy study; improved; in vivo; inhibitor; insight; lead optimization; manufacturing scale-up; mouse model; nanoencapsulated; nanoparticle; natural product inspired; novel; protein degradation; proteostasis; safety study; scale up; screening; small molecule; success; tuberculosis drugs; tuberculosis treatment

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.

项目概要/摘要 项目 1 的中心目标是确定候选 ClpC1 调节剂，用于转化为临床抗- 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将 ClpC1 确定为结核病治疗的新型蛋白质靶标。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，表明 作用于该靶点的结核病药物可用于对抗多重耐药/广泛耐药（MDR/XDR）- 结核病，有可能缩短治疗时间。已知 ClpC1 调节剂的药代动力学 (PK) 特性， ecumicin、rufomycin、cyclomarin-A 和 lassomycin，排除这些 NP 环肽直接用作口服结核病治疗 药物。因此，项目 1 寻求识别和开发新型口服生物可利用的 ClpC1 调节剂，具有潜在的 在本 CETR 设想的结核病治疗方案内使用。潜在客户识别的多种方法包括： 识别新的 NP 和新型小分子 ClpC1 调节剂，并优化已知 NP 的特性。每个 方法将利用肽化学、基于结构的药物设计 (SBDD)、NP 领域的领先技术 发现和基于片段的药物设计。先导化合物的发现和优化将利用四个正交生物物理学 了解与 ClpC1 结合的关键因素的方法：表面等离子共振 (SPR)、X 射线 晶体学、NMR 和冷冻电镜。纳米封装工作旨在优化 ecumicin 的暴露，以充分 探索其在 Mtb 感染小鼠中的功效。 UIC 项目 1 团队具有独特的优势来遵循这些 目标，开创了新的 NP 技术，在药物发现和先导化合物验证方面具有独特的用途， 导致从放线菌中发现了杜鹃素和红霉素。 UIC还开展了大量工作以获得 证明 Mtb ClpC1 蛋白酶脆弱性的机制见解，支持了拟议的 活动。 UIC 进行的研究将有助于推动 ClpC1 转化为抗 Mtb 药物靶点。这 项目 1 的具体目标在 CETR 内也有很强的合作关系。 [目标1]是优化曝光 NP ClpC1 调节剂探索其抗 Mtb 功效。这涉及与 Myongji 的合作（扩大规模 生产）和普林斯顿大学（纳米封装增强体内功效）。纯度和 NP 完整性 寡肽的分析将采用 UIC 的 qNMR 方法。将进行PK和功效研究 带核心 A+B。 [AIM 2] 将采用多种方法（大规模 NP 分离和表征；受 NP 启发的 结构/SAR 指导设计、NMR 筛选）来识别新型和口服可用的 ClpC1 调节剂。数组 方法涉及 UIC 的分子网络、通过与 Eli Lilly、ClpC1 合作进行的基于片段的筛选 项目 2 的功能测定，以及核心 A、肽和药物化学、SBDD、体外 ADME 和 鼠标PK。 [AIM 3] 旨在评估 NP、合成环肽和 ClpC1 结合亲和力的关键因素 通过使用 SPR、NMR、Cryo-EM 和共结晶 X 射线分析来分析小分子。目标3也将优化 目标 2 中出现的线索，随后通过核心 C 进行候选者选择和 IND 支持研究。