Discovering antimicrobials acting against MDR pathogens

发现针对 MDR 病原体的抗菌药物

• 批准号: 10696159
• 负责人: Kim Lewis
• 金额: $ 114.15万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-02 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10696159
• 关键词: Acinetobacter baumannii; Actinobacteria class; Animal Model; Anti-Bacterial Agents; Antibiotics; Antimicrobial Resistance; Bacteria; Binding Proteins; Biodiversity; Biological Assay; Candida albicans; Cell Wall; Cells; Chronic Disease; Cryoelectron Microscopy; Data; Detection; Diffusion; Dose; Encapsulated; Enterobacter; Enterobacteriaceae; Escherichia coli; Eukaryotic Cell; Gram-Negative Bacteria; Growth; In Situ; In Vitro; Incubated; Industry; Infection; Klebsiella pneumoniae; Label; Mammalian Cell; Membrane Proteins; Methods; Micro Electron Diffraction; Microfluidic Microchips; Microfluidics; Modality; Natural Products; Organism; Pseudomonas aeruginosa; Reporter; Resistance; Salmonella typhimurium; Sampling; Sepharose; Septicemia; Signal Transduction; Soil; Sorting; Source; Stains; Staphylococcus aureus; Structure; Testing; Thigh structure; Time; Toxicity Tests; Withdrawal; antimicrobial; cytotoxicity; experience; fighting; high throughput screening; in vivo; liquid chromatography mass spectrometry; multi-drug resistant pathogen; novel; pathogen; priority pathogen; resistance frequency; scaffold; screening

Abstract We are experiencing an antimicrobial resistance crisis (AMR), a direct result of a decline in antibiotic discovery. The WHO designated a list of priority pathogens, and of these, MDR Gram- negative Enterobacteriaceae (E. coli, S. typhimurium, Klebsiella pneumoniae, Enterobacter), Pseudomonas aeruginosa, and Acinetobacter baumannii) are of “critical priority”. These pathogens are the focus of the present proposal aimed at developing a platform for efficient discovery of novel antimicrobials. The field once enjoyed a golden era of discovery, fueled mainly by screening of soil actinomycetes. All major classes of broad-spectrum antibiotics active against Gram-negative pathogens were discovered by the 1960s. Overmining of actinomycetes resulted in the collapse of the discovery platform. Novel antibiotics discovered since then only act against Gram-positive species. We developed methods to access a broader range of bacteria, with a focus on uncultured species that make up 99% of total biodiversity. A number of novel compounds came from this source, including teixobactin, representing a new class of cell- wall acting compounds without detectable resistance (Ling et al., 2015). Teixobactin is undergoing IND-enabling studies; it is also a narrow-spectrum compound. We propose to develop a platform for efficient discovery of novel antimicrobials. The main problem is the enormous background of toxic, and to a lesser extent, known compounds. We hypothesize that the bottleneck of dereplication can be resolved by differential screening that detects the presence of a promising compound prior to dereplication. Using this approach, we recently discovered darobactins that have a novel scaffold and target the essential outer membrane protein BamA (Imai et al., 2019), and several additional novel compounds. In the proposed project, we will develop an ultra-high throughput screen based on encapsulating producing bacteria together with different fluorescently labeled reporters in microdroplets created in a microfluidics device. Our preliminary data show that sorting droplets can be performed at a rate of 106 a day, and leads to detection of producers of desirable antimicrobials. We will evaluate several modalities of this screen, aimed at discovering selective as well as broad-spectrum compounds acting against Gram-negative bacteria; and anti-persister compounds. The platform is likely to be of use to the field of antibiotic discovery. Leads that come out of this screen will be evaluated in vitro and in animal models of infection. Novel leads that come out of this project will be ready to enter IND-enabling studies.

抽象的 我们正在经历抗菌抗性危机（AMR），这是下降的直接结果 抗生素发现。世卫组织指定了优先病原体的清单，其中包括 阴性肠杆菌科（大肠杆菌，鼠伤寒S. 铜绿假单胞菌和鲍曼尼杆菌杆菌是“关键的优先级”。这些 病原体是本提案的重点，旨在开发一个高效的平台 发现新型抗菌剂。该领域曾经享受着一个发现的黄金时代 主要通过筛选土壤放线菌。所有主要类别的广谱抗生素活跃 在1960年代发现了针对革兰氏阴性病原体。放线菌的过度 导致发现平台的崩溃。从那以后发现的新颖抗生素才发现 对革兰氏阳性物种作用。我们开发了访问更广泛范围的方法 细菌，重点是占总生物多样性99％的未养殖物种。许多 新颖的化合物来自该来源，包括电脱角蛋白，代表了一类新的细胞 - 壁作用化合物无可检测的抗性（Ling等，2015）。 Teixobactin是 进行辅助研究；它也是一种狭窄的光谱化合物。我们建议 开发一个有效发现新型抗微生物的平台。主要问题是 有毒的巨大背景和较小程度的已知化合物。我们假设这一点 可以通过检测到的差分筛选来解决瓶颈的瓶颈 在删除之前存在承诺化合物。使用这种方法，我们最近 发现具有新颖脚手架并靶向必需外膜的darobactins 蛋白质巴马（Imai等，2019）和其他几种新型化合物。在提议中 项目，我们将基于封装生产开发一个超高的吞吐量屏幕 细菌以及在A 微流体设备。我们的初步数据表明，可以以速率进行排序液滴 每天106，并导致检测所需的抗菌剂的生产者。我们将评估 该屏幕的几种方式，旨在发现选择性和广谱 作用于革兰氏阴性细菌的化合物；和抗蛋白酶化合物。平台 可能对抗生素发现领域有用。从这个屏幕出来的线索将是 在体外和感染动物模型中进行了评估。从这个项目中出来的小说线索将 准备进入辅助研究。