Targeting the outer membrane protein translocation pathways

靶向外膜蛋白易位途径

• 批准号: 8462111
• 负责人: STEPHEN LORY
• 金额: $ 20.08万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8462111
• 关键词: Animals; Anti-Bacterial Agents; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Bacteria; Biogenesis; Biological; Biological Assay; Blood; Cell physiology; Clinical; Communicable Diseases; Cytoplasm; Development; Drug Evaluation; Drug Targeting; Engineering; Environment; Exhibits; Genome; Gram-Negative Bacteria; Growth; Human; In Vitro; Infection; Lead; Libraries; Lipoproteins; Liquid substance; Luciferases; Lung; Membrane; Membrane Protein Traffic; Membrane Proteins; Microbial Biofilms; Modeling; Mucous body substance; Mus; New England; Organism; Outcome; Pathway interactions; Pharmaceutical Preparations; Phase; Predisposition; Property; Protein translocation; Proteins; Pseudomonas; Pseudomonas aeruginosa; Regulation; Relative (related person); Reporter; Resistance; Respiratory Tract Infections; Serum; Specificity; Testing; Work; antimicrobial; antimicrobial drug; bactericide; base; beta barrel; cell envelope; chemical genetics; cytotoxicity; efflux pump; experience; high throughput screening; inhibitor/antagonist; killings; medical schools; meetings; membrane activity; novel; pathogen; preclinical study; protein transport; resistance mechanism; respiratory; small molecule; small molecule libraries; trafficking

DESCRIPTION (provided by applicant): Gram-negative bacteria are the causative agents a variety of important human infectious diseases. The successful therapy of infections, caused by many of these pathogens is limited by their intrinsic resistance mechanism including the impermeable outer membrane (OM) and the activities of various efflux pumps. In this project, we propose to develop novel antibiotics that do not enter the bacterial cytoplasm, but instead, they act by interfering with the biogenesis of the OM. Two pathways, consisting of the Lol and Bam machineries, are responsible for trafficking of lipoproteins and beta-barrel non-lipidated OM proteins, respectively. These pathways are essential in Pseudomonas aeruginosa and will be targeted for disruption by small molecule inhibitors. Strains of P. aeruginosa were engineered that allow regulation of the key components of the Bam and Lol pathways and carry a luciferase reporter construct responsive to Lol and Bam depletion. These P. aeruginosa test strains will be used to screen compound libraries and inhibitors of OM protein trafficking will be identified. The compounds will be characterized to identify those with maximal killing potency potent, exhibit a broad spectrum against other Gram-negative pathogens, are active in biofilms, serum and respiratory mucus, exhibit low cytotoxicity and potentiate the bactericidal activities of other antibiotics. The protein targets of these active compounds will be indentified using genetic and chemical approaches. The efficacy of each of these compounds alone, or in combination with other antibiotics, in protecting mice against P. aeruginosa colonization will be tested in a murine respiratory infection model. This work could lead to the development of a new class of broad-spectrum inhibitors suitable for therapy of a variety of infections caused by antibiotic- resistant Gram-negative pathogens.

描述（由申请人提供）：革兰氏阴性细菌是各种重要的人类感染性疾病的病因。许多这些病原体引起的感染的成功治疗受其固有抗性机制的限制，包括不可渗透的外膜（OM）和各种外排泵的活性。在这个项目中，我们建议开发出不进入细菌细胞质的新型抗生素，而是通过干扰OM的生物发生来起作用。两种由LOL和BAM机械组成的途径分别负责脂蛋白和β-桶非蛋白质蛋白的运输。这些途径在铜绿假单胞菌中至关重要，并且将被小分子抑制剂造成的目标。对铜绿假单胞菌的菌株进行了设计，可以调节BAM和LOL途径的关键组成部分，并携带荧光素酶记者的构造，对LOL和BAM DEPTETION有反应。这些铜绿假单胞菌测试菌株将用于筛选复合文库，并确定OM蛋白运输的抑制剂。这些化合物将被表征以识别那些具有最大杀伤效力的人，对其他革兰氏阴性病原体具有广泛的范围，在生物膜，血清和呼吸道粘液中活跃，表现出低的细胞毒性，并增强了细菌活性。 其他抗生素。这些活性化合物的蛋白质靶标将使用遗传和化学方法来凹陷。每种化合物中每种化合物的疗效，或与其他抗生素结合使用，可以在鼠呼吸道感染模型中测试小鼠免受铜绿假单胞菌定植的侵害。这项工作可能导致发展新的广谱抑制剂，适合于治疗由抗生素抗生素革兰氏阴性病原体引起的各种感染。