Development of validated probes for the bacterial type III secretion system

开发用于细菌 III 型分泌系统的经过验证的探针

基本信息

批准号：
10405053
负责人：
Victoria Auerbuch Stone
金额：
$ 74.38万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA CRUZ
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10405053
关键词：
Affinity Affinity Chromatography Antibiotic Resistance Antibiotics Automobile Driving Bacteria Bacterial Infections Binding Biochemical Biological Assay Cell physiology Cells Centers for Disease Control and Prevention (U.S.)Cessation of life Chemicals Chlamydia Chlamydia trachomatis Development Dose Drug Targeting ESKAPE pathogens Eukaryotic Cell Evolution Fluorescence Microscopy Future Genes Genetic Growth Health Health Care Costs Healthcare Human Incidence Libraries Life Mammalian Cell Membrane Molecular Target Multi-Drug Resistance Natural Products Needles Panthera leo Pasteurella pseudotuberculosis Peptoids Periodicity Productivity Protein Secretion Proteins Pseudomonas Pseudomonas aeruginosa Pseudomonas aeruginosa infection Public Health Publishing Resistance development Salmonella Salmonella enterica Sampling Structure Structure-Activity Relationship Susceptibility Gene Synthesis Chemistry Testing Therapeutic Toxic effect Type III Secretion System Pathway Universities Virulence Yersinia Yersinia enterocolitica analog antibiotic resistant infections antimicrobial appendage base bioactive natural products biosynthetic product cheminformatics clinically relevant comparative cost counterscreen cytotoxicity enteric pathogen high throughput screening inhibitor member microbiota next generation novel novel therapeutics obligate intracellular parasite opportunistic pathogen pathogen pathogenic bacteria repository resistant strain response scaffold screening small molecule small molecule libraries synthetic peptide systems research tool transcriptome

项目摘要

With increasing incidence of antibiotic resistance, development of new therapies against bacterial pathogens is essential for global public health. The bacterial type III secretion system (T3SS) represents an excellent drug target because it is externally accessible to small molecules and enables virulence of Pseudomonas, Salmonella, Chlamydia, and numerous other important pathogens. We have developed a high throughput screening pipeline to discover T3SS inhibitors and have shown the robustness of our approach through pilot screens identifying three classes of compounds active against the T3SS. We now propose to broaden our scope and screen three unique libraries comprising ~58,000 natural product fractions developed by members of our consortium as well as two commercial synthetic chemical libraries. According to the CDC, every year over 50,000 healthcare-associated Pseudomonas aeruginosa infections occur in the U.S., >6,000 of which are caused by multidrug resistant strains. To identify Pseudomonas T3SS inhibitors and potential future therapeutics, we will carry out the following aims. In Aim 1, we will implement our primary screen and counterscreens to identify natural product fractions and synthetic compounds with specific T3SS inhibitory activity. In Aim 2, we will validate hits identified in Aim 1, using three orthogonal distinct approaches. The identity and structure of bioactive natural products will be determined and initial structure activity analysis performed on identified synthetic scaffolds. Prioritized compounds will be purified or synthesized and evaluated for off target activity, if any, as well as breadth of activity against T3SSs in multiple relevant pathogens. In Aim 3, mode of action will be determined, using parallel genetic and biochemical approaches. This rigorous strategy will provide ~10 T3SS inhibitor chemical probes with identified molecular targets active against P. aeruginosa and potentially other pathogens.

随着抗生素抗性的发生率的增加，针对细菌病原体的新疗法的发展为对于全球公共卫生至关重要。细菌III型分泌系统（T3SS）代表一种优秀的药物目标是因为它在小分子上可以外部访问，并且可以使假单胞菌的毒力具有毒力沙门氏菌，衣原体和许多其他重要病原体。我们已经开发了很高的吞吐量筛选管道以发现T3SS抑制剂，并通过飞行员显示了我们方法的鲁棒性筛选识别针对T3S的三类化合物的三类。我们现在建议扩大我们的范围和屏幕三个独特的库，包括约58,000个由成员开发的天然产品分数我们的财团以及两个商业合成化学库。根据疾病预防控制中心，每年在美国发生了50,000多种医疗保健相关的假单胞菌感染，其中6,000张是由多药抗性菌株引起。识别假单胞菌T3SS抑制剂和潜在的未来治疗剂，我们将执行以下目标。在AIM 1中，我们将实施我们的主要屏幕，尺寸以鉴定具有特定T3SS的天然产物分数和合成化合物活动。在AIM 2中，我们将使用三种正交不同的方法来验证AIM 1中确定的命中。这将确定生物活性天然产物的身份和结构，并初始结构活动分析对已识别的合成支架进行。优先的化合物将被纯化或合成和评估对于多种相关病原体中的T3SS活动（如果有的话），以及针对T3SS的活动广度。目标 3，将使用平行的遗传和生化方法确定作用方式。这个严格策略将提供约10个T3SS抑制剂化学探针，并具有针对P的分子靶标。铜绿和潜在的其他病原体。