Virulence inhibitors as candidate therapeutics in CF

毒力抑制剂作为 CF 的候选治疗方法

• 批准号: 6859662
• 负责人: STEPHEN LORY
• 金额: $ 21.19万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-20 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6859662
• 关键词: CHO cells; Pseudomonas aeruginosa; antibacterial agents; bacterial cytopathogenic effect; bacterial genetics; bacterial proteins; chemical genetics; chemical registry /resource; cystic fibrosis; cytotoxicity; drug discovery /isolation; gene expression; high throughput technology; pharmacokinetics; protein structure function; recombinant proteins; small molecule; transcription factor; virulence

DESCRIPTION (provided by applicant): The major cause of morbidity and mortality of patients with cystic fibrosis is the chronic respiratory infection caused primarily by an opportunistic pathogen Pseudomonas aeruginosa. The bacteria that colonize the respiratory tract of CF patients utilize an armament of virulence factors to overcome the host defense mechanisms directed primarily at polymorphonucelar leukocytes. One such mechanism is production of cytotoxic and anti-phagocytic factors ("effectors"), targeted into the host cells by the type III secretion system (TTSS). The objective of this application is to utilize the new field of chemical genetics to develop small molecules as probes of function of the TTSS. The aims of this project are to utilize a sensitive high throughput bioassay (TTSS dependent killing of cultured CHO cells) to identify small molecule inhibitors of cytotoxicity, and further define the mechanism of inhibition of the specific stages of TTSS expression, effector secretion from P. aeruginosa and their translocation into the mammalian cell. The active compounds that interfere with the action of TTTS will be screened for activity in P. aeruginosa from a variety of clinical sources, with emphasis on mucoid and non-mucoid CF isolates. Furthermore, for each compound or family of compounds, we will identify the specific component of the TTSS, which is the target of inhibition. There is little doubt that there is an urgent need to develop new classes of antimicrobials active against P. aeruginosa during all stages of respiratory track colonization of CF patients. An additional outcome of this project will be the identification of groups of small molecules and the establishment of structure activity relationships among them such that they may serve as lead compounds for the development of therapeutic agents against P. aeruginosa by inhibiting a key virulence mechanism. This project therefore can lay the foundation for a new research direction in the field of TTSS, leading to a better understanding of this important mechanism of effector targeting, which is currently not possible through the more traditional use of mutants in the TTSS components.

描述（由申请人提供）：囊性纤维化患者发病率和死亡率的主要原因是慢性呼吸道感染主要是由铜绿假单胞菌的机会性病原体引起的。 CF患者呼吸道定居的细菌利用毒力因子的武器克服主要针对多形核白细胞的宿主防御机制。一种这样的机制是通过III型分泌系统（TTSS）靶向宿主细胞的细胞毒性和抗吞噬因子（“效应子”）的产生。该应用的目的是利用新的化学遗传学领域发展小分子作为TTSS功能的探针。该项目的目的是利用敏感的高通量生物测定（TTSS依赖性杀害培养的CHO细胞）来识别细胞毒性的小分子抑制剂，并进一步定义了TTSS表达的特定阶段的抑制机制，铜绿假单胞菌及其转运量的效果分泌及其转运量。干扰TTTS作用的活性化合物将从多种临床来源中筛选在铜绿假单胞菌中的活性，重点是粘液和非粘液CF分离株。此外，对于每种化合物或化合物家族，我们将确定TTSS的特定成分，这是抑制的靶标。毫无疑问，迫切需要在CF患者的呼吸道定植的各个阶段开发针对铜绿假单胞菌的新型抗菌剂。该项目的另一个结果将是鉴定小分子组以及建立它们之间的结构活性关系，从而可以通过抑制关键的毒力机制来作为针对铜绿假单胞菌开发治疗剂的铅化合物。因此，该项目可以为TTSS领域的新研究方向奠定基础，从而更好地理解这种重要的效应器靶向机理，这是通过在TTSS组件中更传统地使用突变体的目前不可能的。