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: 10526217
• 负责人: Tanya Parish
• 金额: $ 29.54万
• 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 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10526217
• 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; 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; base; counterscreen; drug discovery; drug-sensitive; global health; inhibitor; interest; macrophage; 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.

总结 结核病仍然是全球主要的健康负担，2019年新增病例1000万， 数十亿人口。结核病的死亡人数现在超过了艾滋病毒（2019年为120万人）。分枝杆菌 结核病是一种复杂的病原体，可在人体内持续数十年 并且需要使用多种抗生素进行长期治疗。 M.结核病是它在人体细胞内生存和复制的能力，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 这可能是很难杀死的，部分原因是分子进入真核细胞的要求， 部分原因是细菌持续存在的不同生理状态。越来越多的证据表明， 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含量筛选的表型筛选方法， 同时监测细菌和真核细胞的存活。我们做了一个初步筛选来鉴定 分枝杆菌细胞内复制的抑制剂。我们发现了几个感兴趣的系列，并选择了三个系列 具有吸引人的物理化学性质。基于我们最初的成功，我们建议扩大我们的努力， 一个更大的多样化筛选集。 该提案的总体目标是筛选针对细胞内Mtb的分子的多样性集，并评估其对细胞内Mtb的抑制作用。 并为未来的工作划分优先级。我们将结合联合收割机的生物活性，概况，作用模式和物理化学 属性来选择最有希望的系列。这一探索性建议的主要成果将是新颖的 化学物质准备进入发现管道。