Modulation of Protein production and Degradation as an integrated approach to rapid sterilization of Drug sensitive and resistant Mtb.

CETR的主要目的是调节蛋白质的产生和降解，作为一种综合方法 对药物敏感（DS）和耐药结核病（DR-TB）进行快速灭菌，最终目标是 提交2份研究性新药（IND）申请和对两种DS均有效的治疗方案 和耐药结核病，并可能导致结核病感染小鼠在2个月或更短时间内无复发治愈，同时还 抑制抗性发展。从历史上看，结核病治疗方案的开发主要是 经验的我们目前的治疗来自数十年来进行的系列临床试验。最近， 我们使用小鼠模型加速了这一经验过程，迄今为止，该模型具有良好的预测能力。 然而，这些方案的任何理由都是事后的--我们真的不明白为什么某些 组合比其他组合更好。在这里，我们将奋进从基本原则设计一个更好的养生法。我们 已知RNA聚合酶（RNAP）抑制在临床上被证明可显著缩短治疗时间， 利福平与多种药物协同作用。通过遗传学研究，我们发现蛋白质降解是一种 特别脆弱的过程，因为即使适度抑制Clp蛋白酶活性也会导致细胞死亡。我们 这是因为在从转录到翻译的“蛋白质稳态”途径中的多重损伤， 蛋白质周转将可能导致更有效的结核病治疗方案。事实上，我们的初步体内数据 表明这是真的。这个多学科的CETR联盟将汇集三个主要领域的关键专业知识 药物靶点构成维持蛋白质稳态的复杂和协调的过程网络， TB.通过这一系列高度相互关联的项目和核心，我们将能够： 通过发现ClpC 1的口服活性调节剂和小分子ClpP 1 P2， 蛋白酶抑制剂（项目1和项目2以及核心A、B和C）。使用结构引导的药物发现 方法和最新的配方技术，这些调制剂将得到优化和改进， 临床前候选人选择。我们希望至少有一个临床前候选人出现在这些不同的 接近。鉴定在非重叠位点结合并抑制酶的新型RNAP抑制剂， 因此对药物敏感性和耐药结核病都有效的利福霉素（项目3和核心A，B， 和C）。使用结构信息和计算化学将指导我们的努力， 选择.使用体外中空纤维系统和小鼠结核分枝杆菌感染模型，我们将表征 控制每种药物的抗TB活性和耐药突变体抑制的PK/PD关系 候选物（ClpC 1调节剂[项目1]、ClpP 1 P2调节剂[项目2]、RNAP抑制剂[项目3]和一种抗肿瘤药物[项目4]）。 更安全的恶唑烷酮已经确定，目前正在IND使能研究中，[项目4]），并提供最佳的 普遍有效的方案。

The main objective of the CETR is to modulate protein production and degradation as an integrated approach to rapid sterilization of drug sensitive (DS) and drug resistant tuberculosis (DR-TB) with the ultimate goal of delivering 2 investigational new drug (IND) applications and a regimen that will be effective against both DS and DR-TB and could result in relapse-free cures of TB-infected mice in 2 months or less, while also suppressing resistance development. Historically, TB treatment regimen development has been largely empiric. Our current treatment arises from serial clinical trials performed over the course of decades. Recently, we have accelerated this empiric process using a mouse model which, thus far, has excellent predictive power. However, any rationale for these regimens is ex post facto - we really do not understand why certain combinations are better than others. Here we will endeavor to devise a better regimen from first principles. We know that inhibition of RNA polymerase (RNAP) is clinically proven to shorten therapy dramatically and that rifampin synergizes with a variety of drugs. Using genetic studies, we have found that protein degradation is a particularly vulnerable process as even modest inhibition of Clp protease activity results in cell death. We reason that multiple insults in the “proteostasis” pathway that leads from transcription through translation and protein turnover will likely result in more potent TB treatment regimens. Indeed, our preliminary in vivo data suggest this is true. This multidisciplinary CETR consortium will bring together key expertise on three major drug targets that constitute the complex and coordinated network of processes that maintain proteostasis in TB. Through this highly interconnected set of projects and cores we will be able to: Identify modulators of the Clp protease complex by discovering an orally active modulator of ClpC1, and a small molecule ClpP1P2 protease inhibitor (Project 1 and Project 2 and Cores A, B and C). Using structure guided drug discovery approaches and the latest formulation technologies these modulators will be optimized and advanced to preclinical candidate selection. We expect at least one preclinical candidate to emerge from these various approaches. Identify novel RNAP inhibitors that bind and inhibit the enzyme at a non-overlapping site than rifamycins that will therefore be effective against both drug-sensitive and DR-TB (Project 3 and Cores A, B, and C). Use of structural information and computational chemistry will guide our effort to preclinical candidate selection. Using an in vitro hollow fiber system and mouse Mtb-infection models, we will characterize the PK/PD relationships that govern the anti-TB activity and suppression of drug-resistant mutants for each drug candidate (ClpC1 modulators [Project 1], ClpP1P2 modulators [Project 2], RNAP inhibitors [Project 3] and a safer oxazolidinone already identified and currently in IND enabling studies, [Project 4]) and deliver the optimal universally active regimen.