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 数十亿人口。目前，结核病死亡超过了艾滋病毒（2019年120万）。分枝杆菌 结核病是一种病原体，是一种复杂的病原体，可以在人类宿主中持续数十年 并且需要长期治疗多种抗生素。 结核分枝杆菌的特征之一是它在人类细胞内生存和复制的能力，包括 巨噬细胞是针对感染的正常宿主防御机制之一。细胞内细菌是特定的 可能很难杀死的人口，部分原因是分子进入真核细胞，而在 部分由于细菌持续存在的生理状态不同。增加证据指向更高的证据 细胞内细菌中抗生素耐受性水平以及异质性增加。 为了找到新代理，我们开发了一种利用高内容筛选的表型筛选方法 同时监测细菌和真核细胞的存活。我们运行了一个试点屏幕以识别新型化学物质 分枝杆菌细胞内复制的抑制剂。我们发现了一些系列兴趣，并选择了三个系列 具有吸引人的理化特性。根据我们的最初成功，我们建议扩大我们的努力 更大的不同筛选集。 该提案的总体目的是筛选针对细胞内MTB的一组分子，并评估 并优先考虑未来工作的热门歌曲。我们将结合生物学活性，轮廓，作用方式和物理化学 选择最有前途的系列的属性。该探索性建议的主要结果将是新颖的 化学物质准备进入发现管道。