Novel Antibacterial Drugs Targeting DNA Repair Enzymes

针对 DNA 修复酶的新型抗菌药物

• 批准号: 7876767
• 负责人: GERALD R SMITH
• 金额: $ 8.8万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-19 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7876767
• 关键词: Adverse effects; Anti-Bacterial Agents; Antibiotics; Bacteria; Bacterial DNA; Bacterial Infections; Bacteriophage T4; Bacteriophages; Biological Assay; Cells; Cessation of life; Collection; Communicable Diseases; Complement; DNA; DNA Damage; DNA Repair; DNA Repair Enzymes; DNA Repair Pathway; DNA Sequence; DNA repair protein; Drug Compounding; Drug Delivery Systems; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Escherichia coli; Eukaryota; Evolution; Exodeoxyribonuclease V; Face; Future; Genes; Genetic; Genetic Recombination; Growth; Helicobacter pylori; Homologous Gene; Human; Hydrogen Peroxide; Induced Mutation; Infection; Knowledge; Lead; Left; Mediating; Mus; Mutation; Outcome; Pharmaceutical Preparations; Phenocopy; Population; Property; Proteins; Research; Resistance; Screening procedure; Single-Stranded DNA; Specificity; Stomach; Testing; Variant; Work; bacterial resistance; base; chemical release; combat; effective therapy; fighting; helicase; high throughput screening; homologous recombination; inhibitor/antagonist; interest; mouse model; mutant; novel; nuclease; pathogen; pathogenic bacteria; prevent; public health relevance; research study; small molecule; small molecule libraries

DESCRIPTION (provided by applicant): Upon bacterial infection, host cells release chemicals that damage the bacterial DNA. Such damage, if left unrepaired, leads to bacterial death. Recombination-based DNA repair proteins, specifically the RecBCD and AddAB helicase-nucleases, are excellent targets for a new class of anti- bacterial agents because these proteins 1) are required for the major pathway of DNA repair and are widely distributed in bacteria but apparently absent from eukaryotes, 2) are specifically required during infection by several diverse pathogens, and 3) contribute to induced mutation that causes resistance to existing antibiotics. Drugs against these enzymes should self-limit evolution of bacterial resistance. We will first screen for small-molecule inhibitors of RecBCD in Escherichia coli cells, using a simple, quick, and sensitive assay of nuclease activity in an easily grown E. coli strain. Interesting small molecules will be further tested for inhibition of RecBCD-mediated DNA repair and homologous recombination in E. coli and inhibition of purified enzyme in nuclease and helicase assays. We will use this same approach to look for inhibitors of AddAB from the important gastric pathogen Helicobacter pylori. We have shown that addAB deletion mutants have impaired colonization ability; thus, inhibiting AddAB activity should limit bacterial infections. This approach should allow us to identify drugs against similar enzymes from other pathogenic bacteria. These drugs should limit evolution of bacterial resistance and allow more effective treatment of infectious diseases. PUBLIC HEALTH RELEVANCE: Our long-term objective is to discover and develop a novel class of antibacterial drugs that will provide a new means to combat bacterial infections in the face of bacterial resistance to many currently used antibiotics. Upon bacterial infection, host cells release chemicals that damage the bacterial DNA. Such damage, if left unrepaired, leads to bacterial death. Recombination-based DNA repair proteins are excellent targets for a new class of anti-bacterial agents because inhibitors of these proteins will cripple DNA repair and lead to bacterial death. Drugs against these enzymes should also limit evolution of bacterial resistance and allow more effective treatment of infectious diseases.

描述（由申请人提供）：细菌感染后，宿主细胞释放破坏细菌DNA的化学物质。这种损伤如果不修复，会导致细菌死亡。基于解旋的DNA修复蛋白，特别是RecBCD和AddAB解旋酶-核酸酶，是一类新的抗菌剂的优良靶标，因为这些蛋白1）是DNA修复的主要途径所需的，并且广泛分布在细菌中，但显然不存在于真核生物中，2）在几种不同病原体的感染期间是特别需要的，和3）有助于引起对现有抗生素的抗性的诱导突变。针对这些酶的药物应该自我限制细菌耐药性的进化。我们将首先在大肠杆菌细胞中筛选RecBCD的小分子抑制剂，使用一种简单、快速和灵敏的核酸酶活性测定法，在一种容易生长的大肠杆菌中进行。大肠杆菌菌株。将进一步测试感兴趣的小分子在大肠杆菌中对RecBCD介导的DNA修复和同源重组的抑制。大肠杆菌中，并在核酸酶和解旋酶测定中抑制纯化的酶。我们将使用相同的方法从重要的胃病原体幽门螺杆菌中寻找AddAB的抑制剂。我们已经表明，addAB缺失突变体具有受损的定殖能力;因此，抑制AddAB活性应该限制细菌感染。这种方法应该使我们能够识别针对其他病原菌类似酶的药物。这些药物应该限制细菌耐药性的演变，并允许更有效地治疗感染性疾病。公共卫生相关性：我们的长期目标是发现和开发一类新的抗菌药物，这将提供一种新的手段来对抗细菌感染，面对细菌对许多目前使用的抗生素的耐药性。在细菌感染时，宿主细胞释放破坏细菌DNA的化学物质。这种损伤如果不修复，会导致细菌死亡。基于降解的DNA修复蛋白是一类新型抗菌剂的极好靶点，因为这些蛋白的抑制剂会削弱DNA修复并导致细菌死亡。针对这些酶的药物也应该限制细菌耐药性的演变，并允许更有效地治疗感染性疾病。