Screen for synthetic lethality in Mycobacterium tuberculosis

结核分枝杆菌综合致死率的筛选

• 批准号: 8063148
• 负责人: WILLIAM Ramses BISHAI
• 金额: $ 4.06万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-15 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8063148
• 关键词: Adjuvant; Amoxicillin; Antibiotics; Biological Assay; Cells; Clarithromycin; Clavulanate; Combined Antibiotics; Emergency Situation; Erythromycin; Evaluation; Extreme drug resistant tuberculosis; Family; Fluorescence; Goals; Imipenem; Laboratories; Lactams; Lead; Libraries; Macrolide Antibiotics; Macrolides; Microbe; Molecular Bank; Molecular Target; Monobactams; Mutation; Mycobacterium tuberculosis; Penetration; Penicillins; Pharmaceutical Preparations; Predisposition; Protocols documentation; Resources; Screening procedure; Specificity; Structure; Tetracyclines; Tuberculosis; Virulent; base; global health; high throughput screening; member; miniaturize; public health relevance; tuberculosis drugs; tuberculosis treatment

DESCRIPTION (provided by applicant): Tuberculosis has been declared a WHO global health emergency, and the recent emergence of multidrug (MDR) and extensively drug resistant (XDR) TB has underscored the urgent need for new TB drugs. We hypothesize that HTS will identify probes which chemically sensitize Mycobacterium tuberculosis (M. tb.) to the activity of existing 2-lactam and macrolide antibiotics. The identification of such probes could lead to combination antibiotics, similar to amoxicillin-clavulanate, which have potency against M. tb. This proposal seeks access to the HTS resources provided by the Molecular Libraries Screening Center Network (MLSCN) with the goal of identifying sensitizing probes, identifying their molecular targets, and finding candidate agents as potential lead structures for further study. An HTS-ready, fluorescence-based whole cell screening assay is proposed that will utilize virulent M. tb. in combination with optimized concentrations of imipenem (IMI, representing the 2-lactam class) and clarithromycin (CLA, representing the macrolide class). The following aims are proposed to conduct an efficient HTS evaluation for probes that are synthetically lethal against M. tb. in the presence of 2-lactam and macrolide antibiotics: to transfer the M. tb./adjuvant drug susceptibility protocol to the designated screening center to reproduce, miniaturize, and automate a corresponding HTS assay that will be used to screen the MLSCN compound library for sensitizing probes, to provide technical support, and to conduct secondary assays confirming the potency and specificity of identified probes. PUBLIC HEALTH RELEVANCE: Tuberculosis has been declared a WHO global health emergency, and the recent emergence of multidrug (MDR) and extensively drug resistant (XDR) TB has underscored the urgent need for new TB drugs; indeed, many antibiotics such as those from the penicillin-, erythromycin-, and tetracycline-families are not clinically useful in the treatment of TB either because of poor bacterial penetration or drug degradation by the microbe. Our preliminary studies reveal that specific mutations can sensitize Mycobacterium tuberculosis (M. tb.) to penicillin and erythromycin, thus supporting the hypothesis that new drugs may be identified that would also sensitize the microbes to the activity of these familiar classes of antibiotics. We have developed a microtiter plate assay to seek compounds which amplify the activity of key members of the penicillin- and the erythromycin-class against M. tb., and we propose to transfer this assay to an appropriate MLSCN laboratory for high-throughput screening to identify potential lead compounds.

描述（由申请人提供）：结核病已被世卫组织宣布为全球突发卫生事件，最近出现的多药（MDR）和广泛耐药（XDR）结核病强调了对新型结核病药物的迫切需求。我们假设HTS将鉴定出使结核分枝杆菌（M. tb.）对现有的2-内酰胺类和大环内酯类抗生素的活性化学致敏的探针。这种探针的鉴定可能导致类似阿莫西林-克拉维酸盐的联合抗生素的产生，这种抗生素对结核分枝杆菌具有效力。本提案寻求通过分子文库筛选中心网络（MLSCN）提供的HTS资源，以确定致敏探针，确定其分子靶点，并寻找候选药物作为进一步研究的潜在先导结构。提出了一种hts就绪、基于荧光的全细胞筛选试验，该试验将利用致病性结核分枝杆菌。与优化浓度的亚胺培南（IMI，代表2-内酰胺类）和克拉霉素（CLA，代表大环内酯类）联合使用。提出以下目标，以便对对结核分枝杆菌具有综合致死作用的探针进行有效的HTS评价。在2-内酰胺和大环内酯类抗生素存在的情况下：转移结核分枝杆菌。/辅助药敏方案到指定的筛选中心进行复制，小型化和自动化相应的HTS分析，该分析将用于筛选MLSCN化合物库的致敏探针，提供技术支持，并进行二次分析，确认鉴定探针的效力和特异性。