Discovering antimicrobials acting against MDR pathogens

发现针对 MDR 病原体的抗菌药物

基本信息

批准号：
10502744
负责人：
Kim Lewis
金额：
$ 113.65万
依托单位：
NORTHEASTERN UNIVERSITY
依托单位国家：
美国
项目类别：
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/10502744
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 GTP-Binding Protein alpha Subunits, Gs Gram-Negative Bacteria Growth In Situ In Vitro Incubated Industry Infection Klebsiella pneumoniae Label Mammalian Cell Membrane Proteins Methods Microfluidic Microchips Microfluidics Modality Natural Products Organism Pseudomonas aeruginosa Reporter Resistance Salmonella typhimurium Sampling Sepharose Septicemia Signal Transduction Soil Sorting - Cell Movement Source Stains Staphylococcus aureus Structure Testing Thigh structure Time Toxicity Tests Withdrawal antimicrobial base cytotoxicity experience fighting high throughput screening in vivo liquid chromatography mass spectrometry multi-drug resistant pathogen novel pathogen priority pathogen resistance frequency scaffold screening

项目摘要

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素耐药性危机 (AMR)，这是抗生素耐药性下降的直接结果世界卫生组织指定了一份优先病原体清单，其中包括耐多药革兰氏阴性菌。阴性肠杆菌科（大肠杆菌、鼠伤寒沙门氏菌、肺炎克雷伯菌、肠杆菌），铜绿假单胞菌和鲍曼不动杆菌）是“关键优先事项”。病原体是本提案的重点，旨在开发一个有效的平台新型抗菌药物的发现曾一度享受着发现的黄金时代。主要通过土壤放线菌筛选所有主要类别的广谱抗生素活性。 20 世纪 60 年代，人们发现了针对革兰氏阴性病原体的过度开采放线菌。导致了自那时以来发现的新型抗生素的崩溃。我们开发了针对更广泛的革兰氏阳性菌的方法。细菌，重点关注占生物多样性总量 99% 的未培养物种。新的化合物来自这个来源，包括代表一类新细胞的teixobactin 没有可检测到的耐药性的壁作用化合物（Teixobactin 是）。正在进行 IND 研究；我们建议它也是一种窄谱化合物。开发一个有效发现新型抗菌药物的平台。有毒化合物以及较小程度的已知化合物的巨大背景。去重复的瓶颈可以通过检测差异筛选来解决最近，我们使用这种方法在去重复之前发现了一种有希望的化合物。发现了具有新型支架并靶向重要外膜的darobactins BamA 蛋白（Imai 等人，2019）以及其他几种新型化合物。项目中，我们将开发基于封装生产的超高通量屏幕细菌与不同荧光标记的生产者一起在微滴中产生我们的初步数据表明，可以以一定的速度对液滴进行分类。每天 106 个，并导致检测出所需抗菌剂的生产商，我们将进行评估。该筛选的几种模式，旨在发现选择性和广谱对抗革兰氏阴性细菌的化合物；和抗持久性化合物。从该筛选中得出的线索可能会用于抗生素发现领域。该项目将在体外和动物感染模型中进行评估。准备好进入 IND 支持研究。