High Throughput Screening for Small Compounds to Inhibit the Expression of Strept

高通量筛选抑制链球菌表达的小化合物

• 批准号: 7515223
• 负责人: Hongmin Sun
• 金额: $ 2.5万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7515223
• 关键词: Affect; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Applications Grants; Attenuated; Bacteria; Bacterial Infections; Biological Assay; Biological Process; Cause of Death; Data; Fibrinogen; Future; Gene Expression; Genes; Growth; Human; Image; Invasive; Kanamycin Resistance; Knockout Mice; Knowledge; Lead; Modeling; Molecular Bank; Mus; Numbers; Pathogenesis; Pattern; Plasminogen; Prevalence; Public Health; Publishing; Reagent; Reporting; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Role; Screening procedure; Streptococcal Infections; Streptococcus; Streptococcus pyogenes; Streptokinase; Structure-Activity Relationship; System; Testing; Time; Transgenic Mice; United States National Institutes of Health; Virulence; Virulence Factors; Work; antimicrobial; base; design; high throughput screening; in vivo; infectious disease treatment; inhibitor/antagonist; killings; novel; pathogen; pressure; promoter; repository; small molecule

DESCRIPTION (provided by applicant): The widespread occurrence of antibiotic resistance among bacteria is causing increasing concern as a major public health threats. Current antibiotics cause death or growth arrest in the target bacteria. As a result, antibiotic use exerts strong selective pressure to favor antibiotic resistant strains. Novel antimicrobial reagents that suppress pathogen virulence without selecting for antibiotic resistance provide a promising alternative approach for treatment of infectious diseases. Group A Streptococcus (GAS) is an important human pathogen affecting millions of people globally each year. The streptokinase (SK) is a major GAS virulence factor that activates human plasminogen. In our previous studies, we have established the streptokinase/plasminogen interaction as a critical factor in GAS pathogenesis. We propose to take advantage of this observation and design novel antimicrobial reagents for the treatment of GAS infection. In the preliminary study, we have screened 55,000 small compounds for inhibitors of SK expression in GAS and 23 candidate hit compounds have been identified. An additional high throughput screen of up to 500,000 more small compounds for SK expression inhibitors is proposed by taking advantage of the NIH Molecular Libraries and Imaging roadmap initiative. A growth based screen will be optimized to use a GAS strain with kanamycin resistance gene under control of SK promoter to screen for small compounds that can inhibit kanamycin resistance expression, which will serve as lead compounds for SK expression inhibitors. In the future, global effects of candidate compounds on GAS gene expression will be studied to provide clues for identification of the targets. Two GAS two-component systems that have been demonstrated to regulate SK expression in published reports will be tested as prime candidate targets. In addition, a number of murine GAS infection models established in our previous studies will be used to elucidate the effects of candidate compounds on GAS virulence in vivo. Data collected from the above studies will further our understanding of the contribution of SK to GAS infection and identify small compounds that can inhibit GAS virulence. As a result, alternative approach to treat bacterial infection by interfering with GAS virulence without unduly introducing selection pressure for resistance can be explored to supplement antibiotic treatment.

描述（由申请人提供）：细菌中广泛存在的抗生素耐药性作为主要的公共卫生威胁引起了越来越多的关注。目前的抗生素会导致目标细菌死亡或生长停滞。因此，抗生素的使用施加了强大的选择压力，有利于抗生素抗性菌株。新型抗菌试剂可抑制病原体毒力而不选择抗生素耐药性，为治疗传染病提供了一种有前途的替代方法。 A 组链球菌 (GAS) 是一种重要的人类病原体，每年影响全球数百万人。链激酶 (SK) 是一种主要的 GAS 毒力因子，可激活人纤溶酶原。在我们之前的研究中，我们已经确定链激酶/纤溶酶原相互作用是 GAS 发病机制的关键因素。我们建议利用这一观察结果设计新型抗菌试剂来治疗 GAS 感染。在初步研究中，我们筛选了 55,000 种 GAS 中 SK 表达抑制剂的小化合物，并鉴定了 23 种候选命中化合物。通过利用 NIH 分子库和成像路线图计划，提议对多达 500,000 种用于 SK 表达抑制剂的小化合物进行额外的高通量筛选。基于生长的筛选将被优化，以使用在SK启动子控制下具有卡那霉素抗性基因的GAS菌株来筛选可以抑制卡那霉素抗性表达的小化合物，其将作为SK表达抑制剂的先导化合物。未来将研究候选化合物对GAS基因表达的整体影响，为靶点的识别提供线索。已在已发表的报告中证明可以调节 SK 表达的两种 GAS 双组分系统将作为主要候选目标进行测试。此外，我们之前的研究中建立的一些小鼠GAS感染模型将用于阐明候选化合物对体内GAS毒力的影响。从上述研究中收集的数据将进一步了解 SK 对 GAS 感染的贡献，并鉴定可以抑制 GAS 毒力的小化合物。因此，可以探索通过干扰 GAS 毒力来治疗细菌感染的替代方法，而不过度引入耐药性选择压力，以补充抗生素治疗。