Chemical probes of vulnerable pathways in antibiotic-resistant pathogens

抗生素抗性病原体中脆弱途径的化学探针

基本信息

批准号：
8810639
负责人：
Tanya Parish
金额：
$ 74.48万
依托单位：
ACCESS TO ADVANCED HEALTH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8810639
关键词：
Affinity Chromatography Anti-Bacterial Agents Antibiotic Resistance Antibiotics Azoles Bacillus (bacterium)Binding Cell Death Cell Extracts Cell physiology Cells Cessation of life Chemicals Communicable Diseases Cytochrome P450 Detection Drug Targeting Drug resistance Drug resistance in tuberculosis Enzymes Essential Genes Expression Library Free Radicals Future Generations Genetic Genetic Engineering Growth Human Hybrids Immobilization Individual Label Lead Libraries Life Mass Spectrum Analysis Measures Metabolic Metabolism Methods Molecular Multi-Drug Resistance Mutagenesis Mutation Mycobacterium tuberculosis Outcome Pathogenesis Pathway interactions Pharmaceutical Preparations Photoaffinity Labels Process Protein Binding Proteins Reactive Oxygen Species Reagent Recombinants Reporter Genes Reporting Resistance Resources Series Single Nucleotide Polymorphism Staphylococcus aureus Structure-Activity Relationship System Techniques Testing Tuberculosis Work Yeasts analog base cellular targeting chemical property cytotoxicity drug development drug discovery genetic approach genome sequencing inhibitor/antagonist killings mutant novel pathogen research study resistant strain small molecule tuberculosis treatment

项目摘要

DESCRIPTION (provided by applicant): Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), is a pathogen of global importance; 9 million new cases of TB are reported annually, with 1-2 million deaths. There are half a million cases of drug resistant TB cases each year and the problem of antibiotic resistance is rising. TB treatment is a lengthy process (6-24 months) giving rise to an urgent need for newer, more effective antibiotics. In order to develop new drugs, we need to understand which metabolic processes are the most important for bacterial survival and pathogenesis. Chemical entities can be powerful probes for the identification of key cellular processes underpinning survival. However, most recent work has focused on genetic approaches to identifying essential genes and there has been little progress in identifying the pathways that effective anti-bacterial compounds target. Recently, a number of novel compounds with unknown targets have been identified with cidal or static activity against drug-resistant pathogens including M. tuberculosis. We propose to apply a combination of techniques in conjunction with several compound classes in order to identify vulnerable pathways, which can be thoroughly characterized using compounds as chemical probes. We will use three main approaches to characterize anti-tubercular agents. (1) We will isolate compound-resistant strains (spontaneous mutants or over-expressing recombinant strains) to identify the protein targets and mode of resistance. (2) We will use three methods (affinity chromatography, photoaffinity labeling, and yeast three hybrid system) to identify protein targets which bind to the compounds. (3) We will look at the effect of compound exposure on cell metabolites and the induction of reactive oxygen species. Together these methods will enable us to identify the pathways and specific proteins that are targeted by each compound; such targets will form the basis for future drug discovery and development.

描述（由申请人提供）：结核分枝杆菌是结核病（TB）的病原体，是一种全球重要的病原体；每年报告 900 万新发结核病例，其中 1-200 万人死亡。每年有五十万例耐药结核病病例，抗生素耐药性问题日益严重。结核病治疗是一个漫长的过程（6-24 个月），因此迫切需要更新、更有效的抗生素。为了开发新药，我们需要了解哪些代谢过程对于细菌的生存和发病机制最重要。化学实体可以成为识别支撑生存的关键细胞过程的强大探针。然而，最近的工作主要集中在识别必需基因的遗传方法上，而在识别有效抗菌化合物的靶向途径方面进展甚微。最近，许多具有未知靶点的新型化合物已被鉴定出对包括结核分枝杆菌在内的耐药病原体具有杀灭或静态活性。我们建议将多种技术与几种化合物类别结合应用，以识别脆弱的途径，可以使用化合物作为化学探针来彻底表征这些途径。我们将使用三种主要方法来表征抗结核药物。 (1)分离化合物耐药菌株（自发突变体或过表达重组菌株），鉴定其靶蛋白和耐药模式。 (2)我们将使用三种方法（亲和层析、光亲和标记和酵母三混合系统）来鉴定与化合物结合的蛋白质靶标。 (3) 我们将研究化合物暴露对细胞代谢物和活性氧诱导的影响。这些方法将使我们能够识别每种化合物的靶向途径和特定蛋白质；这些目标将构成未来药物发现和开发的基础。