Validation and development of trans-translation as a drug target for MRSA

反式翻译作为 MRSA 药物靶标的验证和开发

• 批准号: 8704009
• 负责人: KENNETH C KEILER
• 金额: $ 20.86万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8704009
• 关键词: Adverse effects; Antibiotics; Bacteria; Binding; Biological Assay; Cells; Collaborations; Complex; Data; Development; Disease; Drug Targeting; Drug resistance; Eukaryota; Future; Genetic; Goals; Growth; Human; Infection; Institutes; Lead; Luciferases; Modeling; Mutagenesis; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Phase; Reagent; Reporter; Research; Resistance; Ribosomes; Role; Scientist; Staphylococcus aureus; Testing; Toxicity Tests; Translations; Validation; Virulence; World Health; base; cell growth; drug candidate; drug development; high throughput screening; improved; in vivo; in vivo Model; inhibitor/antagonist; methicillin resistant Staphylococcus aureus; novel; prevent; public health relevance; research study; resistant strain; screening; small molecule; tmRNA

DESCRIPTION (provided by applicant): The continued spread of Staphylococcus aureus strains that are resistant to available antibiotics has created a severe problem for treating S. aureus infections. One of the critical impediments to developing drugs that are effective against these deadly MRSA strains is the lack of new antibiotic targets. The trans-translation pathway for resolving stalled ribosomes is a potential target for drug development because it is found in all bacteria and is required for growth of MRSA in culture. The long-term goal of this project is t identify specific inhibitors of trans-translation in S. aureus that can be used as lead compounds for drug development. The overall objective of this proposal is to validate trans-translation as a viable antibiotic target in S. aureus and to develop a primary assay compatible with high-throughput screening (HTS) and secondary assays that can be used to identify inhibitors of trans-translation in S. aureus. The rationale that underlies the proposed research is that validation of trans-translation in MRSA as a drug target and development of assays for HTS will lead to rapid identification of new antibiotics that will dramatically improve treatment for MRSA. The specific aims of this proposal are to confirm that tmRNA and SmpB are essential for S. aureus growth in culture and during infection, to develop primary and secondary screening assays to identify inhibitors of trans-translation in S. aureus, to conduct a pilot screen for tmRNA-SmpB inhibitors, and to use established ex vivo and in vivo assays to characterize hits from screening. Depletion strains and small molecule inhibitors will be used to remove trans-translation activity in during growth in culture and during infection. The results of these experiments will provide the basis for understanding the importance of trans-translation in S. aureus as well as determining if this pathway can be targeted for drug development. An established HTS assay and tmRNA-SmpB activity assays will be adapted for use with S. aureus components. Results from these studies will allow future HTS and high-level optimization of drug candidates. By targeting a pathway that has not been used for antibiotic development, this project will yield new compounds that can be used individually or in combination with existing MRSA therapies.

描述（由申请人提供）：对可用抗生素具有抗药性的金黄色葡萄球菌菌株的持续扩散为治疗金黄色葡萄球菌感染带来了严重的问题。针对这些致命的MRSA菌株有效开发药物的关键障碍之一是缺乏新的抗生素靶标。解决停滞核糖体的跨翻译途径是药物发育的潜在靶标，因为它是在所有细菌中发现的，并且是MRSA在培养中的生长所必需的。该项目的长期目标是确定金黄色葡萄球菌中跨翻译的特定抑制剂，可以用作药物开发的铅化合物。该提案的总体目的是在金黄色葡萄球菌中验证跨翻译为可行的抗生素靶标，并开发与高通量筛选（HTS）兼容的主要测定法（HTS）和次要测定，可用于识别跨跨翻译的抑制剂在金黄色葡萄球菌中。拟议研究基础的基本原理是，将MRSA作为药物靶向的跨翻译验证和HTS测定法的开发将导致对新抗生素的快速鉴定，这些抗生素将大大改善MRSA的治疗方法。该提案的具体目的是确认TMRNA和SMPB对于金黄色葡萄球菌在培养和感染过程中至关重要，以开发主要和次要筛选测定法，以鉴定在金黄色葡萄球菌中进行跨跨性的抑制剂，以进行型号，以进行试验筛查用于TMRNA-SMPB抑制剂，并使用已建立的离体和体内测定来表征筛选的命中。耗竭菌株和小分子抑制剂将用于去除培养和感染期间生长过程中的跨翻译活性。这些实验的结果将为理解金黄色葡萄球菌中反式翻译的重要性提供基础，并确定该途径是否可以针对药物开发。已建立的HTS分析和TMRNA-SMPB活性测定法将适用于金黄色葡萄球菌组件。这些研究的结果将使未来的HT和候选药物的高级优化。通过针对尚未用于抗生素开发的途径，该项目将产生可单独使用或与现有MRSA疗法结合使用的新化合物。