Inhibition of the Bacterial LexA Repressor-Protease to Halt SOS Response-Mediated Resistance and Biofilm Formation

抑制细菌 LexA 阻遏蛋白蛋白酶以阻止 SOS 反应介导的耐药性和生物膜形成

• 批准号: 10194343
• 负责人: Ana V Cheng
• 金额: $ 3.64万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-15 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10194343
• 关键词: Address; Affinity; Agriculture; American; Antibiotic Resistance; Antibiotics; Area; Bacteria; Bacterial Infections; Binding; Binding Sites; Biological; Biological Testing; Bypass; Cell division; Cells; Cessation of life; Cleaved cell; Communities; Contracts; DNA; DNA Damage; Developed Countries; Digestion; Dimethyl Sulfoxide; Drug Targeting; Electrons; Elements; Escherichia coli; Evaluation; Exposure to; Genes; Genetic Transcription; Genotoxic Stress; Gentian Violet; Health; Health Care Costs; Horizontal Gene Transfer; Hospitals; Infection; Interdisciplinary Study; Investigation; Knowledge; Lead; Libraries; Measures; Mediating; Medical; Medical Device; Medicine; Metabolism; Microbe; Microbial Biofilms; Modification; Monitor; Mutagenesis; Nosocomial Infections; Operative Surgical Procedures; Organ; Patients; Peptide Fragments; Peptide Hydrolases; Pesticides; Pharmacologic Substance; Process; Promoter Regions; Proteins; Pseudomonas aeruginosa; Rec A Recombinases; Research; Resistance; Resistance development; SOS Response; Serial Passage; Shelter facility; Son of Sevenless Proteins; Stains; Staphylococcus aureus; Structure; Structure-Activity Relationship; Surface; Transcriptional Regulation; Work; analog; bacterial resistance; chronic infection; clinically relevant; confocal imaging; design; functional group; high throughput screening; improved; in vitro testing; infectious disease treatment; inhibitor/antagonist; interdisciplinary approach; man; multidrug tolerance; novel; novel strategies; pathogen; pressure; prevent; response; sensor; small molecule; small molecule inhibitor; targeted treatment; weapons

Project Summary/Abstract The overuse and misuse of antibiotics has put evolutionary pressure on bacteria to alter or bypass the targets of drugs or otherwise develop resistance, rendering a large percentage of our available medicines and pesticides ineffective. Novel antibiotics have afforded temporary relief due to quick development of resistance, although several pharmaceutical companies have withdrawn from this area of research. Bacterial biofilms further complicate treatment of many bacterial infections. These cell conglomerates contribute to a variety of health conditions and are known to colonize the surfaces of most medical devices. Moreover, they shelter high numbers of persister cells— “dormant” cells which are non-growing and tolerant of most antibiotics. Unfortunately, most existing therapies target metabolic processes which are suspended in these transient subpopulations of bacteria. Altogether we are facing a perfect storm of resistance and tolerance which threatens to kill millions and unravel our current approach to medicine in the process, unless we find a radical solution. To this end, we have identified a potential antibiotic target—the bacterial SOS response. This response to genotoxic stress is conserved across bacteria and has been connected to resistance and tolerance mechanisms, including horizontal gene transfer, mutagenesis, and cell division arrest. Transcription of SOS genes is suppressed by the repressor-protease LexA, which cleaves upon interaction with filamentous protein RecA* to expose the SOS promoter region. A previous high throughput screen identified a potent inhibitor of LexA cleavage. We propose a study to improve this inhibitor and better understand its action and effects. Using a preliminary structure-activity relationship (SAR) study as a guide, we have designed a library of 22-25 analogs for a more in-depth SAR campaign, including analogs specifically designed to overcome potential efflux challenges. Additionally, we have proposed peptide fragments with covalent traps to mimic the native substrate of the LexA protease and irreversibly inhibit its function. Using our most potent inhibitors, we will investigate the downstream biological effects of LexA inhibition, including acquired antibiotic resistance and biofilm formation. We also plan to use photoaffinity probes to identify the inhibitor binding site and orientation within the protein. The uniquely interdisciplinary approach of this proposal will elucidate the mechanism of these inhibitors and will lay the groundwork for a novel strategy to address the resistance and tolerance crisis.

项目摘要/摘要 过度使用和滥用抗生素给细菌带来了进化压力，以改变 或绕过药物的靶标或以其他方式产生抗药性，使很大一部分 我们可用的药物和农药无效。新颖的抗生素暂时提供了 由于阻力的快速发展，尽管几家制药公司已经 撤回了这一研究领域。细菌生物膜进一步复杂化了许多 细菌感染。这些细胞集团有助于各种健康状况，并且是 已知可以定居大多数医疗设备的表面。此外，它们占据了大量的 迫害细胞 - “休眠”细胞，这些细胞是非生长且耐受性大多数抗生素的细胞。 不幸的是，大多数现有疗法针对这些暂停的代谢过程 细菌的瞬态亚群。我们总共面临着一个完美的抵抗风暴， 宽容威胁要杀死数百万的宽容，并在我们目前的医学方法 过程，除非我们找到一种自由基解决方案。 为此，我们已经确定了潜在的抗生素靶标 - 细菌SOS反应。 这种对遗传毒性应激的反应在细菌之间保存下来，并已连接到 耐药性和耐受性机制，包括水平基因转移，诱变和细胞 师逮捕。 SOS基因的转录被复制蛋白蛋白酶lexa抑制 与丝状蛋白质相互作用时裂解以暴露SOS启动子区域。一个 先前的高吞吐量屏幕确定了Lexa裂解的潜在抑制剂。我们提出了一个 研究以改善这种抑制剂并更好地了解其作用和效果。使用初步 结构活性关系（SAR）研究作为指导，我们设计了一个22-25个类似物的库 要进行更深入的SAR运动，包括专门设计的类似物 排出挑战。此外，我们提出了具有共价陷阱的肽片段 Lexa蛋白酶的天然底物和不可逆地抑制其功能。使用我们最有力的 抑制剂，我们将研究Lexa抑制的下游生物学作用，包括 获得的抗生素耐药性和生物膜形成。我们还计划使用光性问题 识别抑制剂结合位点和蛋白质内的取向。独特的跨学科 该提案的方法将阐明这些抑制剂的机制，并将建立 解决抵制和宽容危机的新策略的基础。

项目成果

Exploration of inhibitors of the bacterial LexA repressor-protease.

• DOI: 10.1016/j.bmcl.2022.128702
• 发表时间: 2022-06-01
• 期刊: BIOORGANIC & MEDICINAL CHEMISTRY LETTERS
• 影响因子: 2.7
• 作者: Jaramillo, Ana Victoria Cheng;Cory, Michael B.;Li, Allen;Kohli, Rahul M.;Wuest, William M.
• 通讯作者: Wuest, William M.