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.

项目摘要/摘要 项目1的核心目的是识别候选CLPC1调节剂，以转化为临床抗 - 结核病（TB）药物。正如NIAID总监最近强调的那样，迫切需要新的口服结核病药物。 我们的工作和其他工作已将CLPC1确定为结核病治疗的新型蛋白质靶标。已知的天然产品 （NP）CLPC1调节剂显示出在体外抗肌分杆菌（MTB）特性方面的有利 对该靶标作用的结核病药物将对多药/广泛抗药性（MDR/XDR）具有实用性 - 结核病，有可能缩短治疗。已知CLPC1调节剂的药代动力学（PK）特性， 海丝蛋白，rofomycin，cylomarin-A和lassomycin，排除了这些NP环状宠物的直接使用作为口服TB 毒品。那就是，项目1旨在识别和开发具有潜力的新型口服生物利用的CLPC1调节剂 在此CERTER中设想的结核病治疗方案中使用。铅识别的多种方法包括： 鉴定新的NP和新型的小分子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）的前缘技术，NP 发现和基于碎片的药物设计。铅发现和优化将利用四个正交生物物理 理解与CLPC1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，NMR和冷冻EM。纳米囊化工作旨在优化海丝蛋白的暴露，以完全 探索其在MTB感染的小鼠中的效率概况。 UIC项目1团队的独特位置可以遵循这些 目标，使新的NP技术具有独特的药物发现和铅验证的实用性 从放线菌，海丝蛋白和鲁霉素的发现中发现了这一发现。 UIC还进行了广泛的工作以获得 机械洞察证明了MTB CLPC1蛋白酶的脆弱性，并为提议的基础 活动。在UIC进行的研究将有助于推动CLPC1作为抗MTB药物靶标的翻译。这 项目1的具体目标在CERTR内也具有牢固的协作关系。 [AIM 1]是优化 NP CLPC1调节器探索其抗MTB效率。这涉及与Myongji的合作（扩展 生产）和普林斯顿大学（用于增强体内效率的纳米封装）。纯度和NP完整性 寡肽的分析将采用UIC的QNMR方法。 PK和效率研究将进行 w/cores a+b。 [AIM 2]将采用潜水员方法（大规模的NP隔离和表征；由NP启发 结构/SAR引导的设计，NMR筛选），以识别新颖和口服的CLPC1调节剂。数组 方法涉及在UIC上通过与Lilly，CLPC1合作的UIC分子网络 使用项目2的功能测定，以及在核心A，胡椒和医学化学，SBDD，体外ADME和 鼠标PK。 [AIM 3]试图评估NP，合成环状肽和 使用SPR，NMR，Cryo-EM和共结晶X射线分析的小分子。 AIM 3也将优化 从AIM 2出现的引线，然后通过核心C进行候选和辅助研究。