Project 1 UIC Targeting Protein Degradation ClpC1 ATPase

项目 1 UIC 靶向蛋白质降解 ClpC1 ATPase

• 批准号: 10388412
• 负责人: Scott G. Franzblau
• 金额: $ 85.09万
• 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/10388412
• 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; base; biophysical techniques; candidate selection; design; drug candidate; drug discovery; drug-sensitive; efficacy study; improved; in vivo; inhibitor; insight; lead optimization; mouse model; nanoencapsulated; nanoparticle; 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.

项目总结/摘要 项目1的中心目标是鉴定候选ClpC 1调节剂，用于翻译成临床抗- 结核病药物。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ClpC 1作为结核病治疗的新蛋白质靶点。已知天然产物 (NP)ClpC 1调节剂显示有利的体外抗结核分枝杆菌（Mtb）性质，表明 作用于这一靶标的结核病药物将可用于治疗多药耐药/广泛耐药（MDR/XDR）- 结核病，有可能缩短治疗时间。已知ClpC 1调节剂的药代动力学（PK）性质， ecumicin、rufomycin、cyclomarin-A和lassomycin排除了这些NP环肽作为口服TB的直接使用 毒品因此，项目1寻求鉴定和开发新的口服生物可利用的ClpC 1调节剂，其具有潜在的 在本CETR中设想的结核病治疗方案中使用。潜在客户识别的不同方法包括： 鉴定新的NP和新的小分子ClpC 1调节剂，并优化已知NP的性质。每个 该方法将利用肽化学，基于结构的药物设计（SBDD），NP 发现和基于片段的药物设计。电极导线发现和优化将利用四个正交生物物理 了解与ClpC 1结合的关键因素的方法：表面等离子体共振（SPR），X射线 晶体学核磁共振和冷冻电镜纳米胶囊化工作旨在优化ecumicin的暴露，以充分 探索其在结核分枝杆菌感染小鼠中的功效概况。UIC项目1团队的独特定位是遵循这些 目标，开创了新的NP技术，在药物发现和先导验证方面具有独特的实用性， 导致从放线菌中发现了ecumicin和rufomycin UIC还开展了广泛的工作， 机制的见解，证明Mtb ClpC 1蛋白酶的脆弱性，支持提出的 活动在UIC进行的研究将有助于推动ClpC 1作为抗结核药物靶标的翻译。的 项目1的具体目标在CETR内部也有很强的合作关系。[AIM 1]是为了优化暴露 NP ClpC 1调节剂以探索其抗Mtb功效。这涉及与Myongji的合作（扩大规模 生产）和普林斯顿大学（用于增强体内功效的纳米封装）。纯度和NP完整性 寡肽的分析将采用UIC的qNMR方法。将进行PK和疗效研究 带芯A+B。[AIM 2]将采用不同的方法（大规模NP隔离和表征; NP启发 结构-/SAR-指导设计，NMR筛选），以鉴定新的和口服可利用的ClpC 1调节剂。阵列 的方法涉及UIC的分子网络，通过与Eli Lilly合作进行的基于片段的筛选，ClpC 1 项目2的功能测定，核心A，肽和药物化学，SBDD，体外ADME，以及 小鼠PK。[AIM 3]试图评估NPs、合成环肽和合成环肽的ClpC 1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。Aim 3还将优化 目标2中出现的线索，然后通过核心C进行候选人选择和IND赋能研究。