Species selective dipeptide inhibitors for Mtb proteasome

Mtb 蛋白酶体的物种选择性二肽抑制剂

基本信息

批准号：
8510791
负责人：
Gang Lin
金额：
$ 25.35万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-02-01 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8510791
关键词：
ATP phosphohydrolase Abbreviations Anti-Infective Agents Antibiotics Bacteria Bioavailable Biochemical Biological Assay Biological Availability Cells Cessation of life Chemicals Client Collaborations Complement Complex Core Facility Crystallization Degradation Pathway Development Dipeptides Drug Kinetics Enzymes Genus Mycobacterium Goals Growth Hereditary Disease Heterogeneity Human In Vitro Inhibitory Concentration 50 Lead Libraries Ligase Mammals Metabolic Microsomes Modeling Mus Mycobacterium tuberculosis Nitric Oxide Outcome Oxygen Peptide Hydrolases Pharmaceutical Chemistry Pharmaceutical Preparations Play Population Predisposition Proteasome Inhibition Proteins Research Institute Resistance Rifampin Role Serum Starvation Sterilization Structure System Testing Therapeutic Translating Tuberculosis Ubiquitin Like Proteins bactericide base design feeding in vitro activity in vivo inhibitor/antagonist isoniazid killings multicatalytic endopeptidase complex mycobacterial nitrosative stress peptidomimetics protein degradation public health relevance public health research scale up screening stability testing structural biology tuberculosis drugs

项目摘要

DESCRIPTION (provided by applicant): In vitro activities do not always translate into in vivo activities, and this is true in Mycobacterium tuberculosis (Mtb) therapeutics. For example, the two first line anti-Mtb drugs isoniazid and rifampicin kill Mtb rapidly in vitro, but their steriliation abilities are slowed and reduced in vivo. Heterogeneity of Mtb populations imparts varied susceptibility to the drugs. In particular, slowly-replicating or non-replicating (collectively "NR) Mtb is non-heritably resistant to most first line anti-TB drugs. Our long-term goal is to develop anti-Mtb drugs that kill NR Mtb populations to complement drugs that kill replicating Mtb populations. During the past eight years, components of a prokaryotic ubiquitin-like protein (Pup)-proteasome system have been discovered. Although the Mtb proteasome is dispensable under standard growth conditions, genetic evidence demonstrates its essentiality for Mtb to survive in mice. We also established the concept that despite the essential role of the proteasome in mammals, small chemical molecules can be discovered with extensive (>1000-fold) species selectivity for inhibiting the Mtb proteasome over the human proteasome, and that such inhibition leads to killing of NR Mtb. In other bacteria, disruption of another regulated protein degradation machine, Clp, has led to killing. Thus, either inhibition or forced activation f chambered proteases may represent a new anti-infective strategy. In our recent screening of 1600 capped dipeptides, we identified a lead compound that potently and species-selectively inhibited the Mtb proteasome over the human proteasome. The inhibitors were bactericidal for non-replicating Mtb in synergy with nitric oxide. Using a competition assay, I confirmed that the dipeptides penetrate the mycobacteria and inhibit the proteasome within them. In this application, we will expand our lead compounds into a small focused compound library whose design and synthesis are guided by substrate-profiling, structural analysis, mycobactericidal activity, and metabolic stability.

描述（由申请人提供）：体外活性并不总是转化为体内活性，这在结核分枝杆菌（Mtb）治疗中确实如此。例如，异烟肼和利福平这两种一线抗Mtb药物在体外能迅速杀灭Mtb，但在体内杀菌能力却减慢和降低。结核分枝杆菌群体的异质性导致对药物的敏感性不同。特别是，缓慢复制或非复制（统称为“NR”）Mtb 对大多数一线抗结核药物具有非遗传性耐药性。我们的长期目标是开发杀死 NR Mtb 群体的抗 Mtb 药物以补充药物在过去的八年里，虽然 Mtb 蛋白酶体已被发现，但原核泛素样蛋白 (Pup)-蛋白酶体系统的成分已被发现。在标准生长条件下是可有可无的，遗传证据证明了 Mtb 在小鼠体内生存的重要性。我们还建立了这样的概念：尽管蛋白酶体在哺乳动物中发挥着重要作用，但小化学分子可以以广泛（> 1000 倍）的物种选择性来发现。抑制人类蛋白酶体上的 Mtb 蛋白酶体，并且这种抑制会导致 NR Mtb 的死亡。在其他细菌中，另一种受调节的蛋白质降解机器 Clp 的破坏会导致杀死。或强制激活有室蛋白酶可能代表一种新的抗感染策略。在我们最近对 1600 个加帽二肽的筛选中，我们鉴定了一种先导化合物，它能够有效地、物种选择性地抑制 Mtb 蛋白酶体，而不是人类蛋白酶体。这些抑制剂与一氧化氮协同作用，对非复制型结核分枝杆菌具有杀菌作用。通过竞争测定，我证实二肽可以渗透分枝杆菌并抑制其中的蛋白酶体。在此应用中，我们将把我们的先导化合物扩展为一个小型的集中化合物库，其设计和合成以底物分析、结构分析、分枝杆菌活性和代谢稳定性为指导。