High content, high throughput, diversity screen for novel anti-tubercular agents targeting intracellular bacteria

针对细胞内细菌的新型抗结核药物的高含量、高通量、多样性筛选

• 批准号: 10621242
• 负责人: Tanya Parish
• 金额: $ 24.61万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-12 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10621242
• 关键词: Address; Antibiotics; Antitubercular Agents; Bacteria; Bacterial Infections; Biological; Biological Assay; Cell Survival; Cells; Cessation of life; Chemicals; Communicable Diseases; Data; Development; Diversity Library; Drug resistance in tuberculosis; Eukaryotic Cell; Evaluation; Future; Generations; HIV; Heterogeneity; Host Defense Mechanism; Human; Infection; Lead; Location; Macrophage; Methods; Monitor; Mycobacterium tuberculosis; New Agents; Outcome; Penetration; Persons; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Physiological; Population; Property; Regimen; Running; Series; Time; Tuberculosis; Work; antibiotic tolerance; axenic culture; counterscreen; drug discovery; drug-sensitive; global health; inhibitor; interest; mycobacterial; new therapeutic target; novel; novel therapeutics; pathogen; screening; small molecule libraries; success; targeted agent; tuberculosis drugs; tuberculosis treatment

Summary Tuberculosis remains a major global health burden with 10 million new cases in 2019 and a latently-infected population of billions. Deaths from tuberculosis now exceed those from HIV (1.2 million in 2019). Mycobacterium tuberculosis, the causative agent, is a sophisticated pathogen which can persist for decades in the human host and which requires lengthy treatment for cure with multiple antibiotics. One of the features of M. tuberculosis is its ability to survive and replicate inside human cells, including macrophages, one of the normal host defense mechanisms against infection. Intracellular bacteria are a specific population which can be hard to kill, in part due to the requirement that molecules enter eukaryotic cells, and in part due to the different physiological state in which the bacteria persist. Increasing evidence points to a higher level of antibiotic tolerance in intracellular bacteria, as well as increased heterogeneity. In order to find new agents, we developed a phenotypic screening method utilizing high content screening to monitor bacterial and eukaryotic cell survival simultaneously. We ran a pilot screen to identify novel chemical inhibitors of mycobacterial intracellular replication. We found several series of interest and selected three series with attractive physicochemical properties. Based on our initial success we propose to expand our effort to run a larger diverse screening set. The overall aim of this proposal is to screen a diversity set of molecules against intracellular Mtb and to evaluate and prioritize hits for future work. We will combine biological activity, profile, mode of action and physicochemical properties to select series with the most promise. The major outcome of this exploratory proposal will be novel chemical matter ready to enter the discovery pipeline.

概括 结核病仍然是全球主要的健康负担，2019 年新增病例 1000 万，潜伏感染病例 数十亿人口。结核病死亡人数现已超过艾滋病毒死亡人数（2019 年为 120 万人）。分枝杆菌 结核病的病原体是一种复杂的病原体，可以在人类宿主体内持续数十年 并且需要使用多种抗生素进行长期治疗才能治愈。 结核分枝杆菌的特征之一是它能够在人体细胞内生存和复制，包括 巨噬细胞，正常宿主防御感染的机制之一。细胞内细菌是一种特殊的 很难杀死的种群，部分原因是分子进入真核细胞的要求，并且在 部分原因是细菌存在的不同生理状态。越来越多的证据表明更高 细胞内细菌的抗生素耐受性水平以及异质性增加。 为了寻找新的药物，我们开发了一种利用高内涵筛选的表型筛选方法 同时监测细菌和真核细胞的存活。我们进行了试点筛选来识别新型化学物质 分枝杆菌细胞内复制的抑制剂。我们找到了几个感兴趣的系列并选择了三个系列 具有有吸引力的物理化学性质。基于我们最初的成功，我们建议扩大我们的努力 更大的多样化筛选集。 该提案的总体目标是筛选一组针对细胞内 Mtb 的多样性分子并评估 并优先考虑未来工作的热门内容。我们将结合生物活性、概况、作用方式和理化 属性来选择最有前途的系列。这个探索性提案的主要成果将是新颖的 化学物质已准备好进入发现管道。