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）：金黄色葡萄球菌菌株对现有抗生素耐药的持续传播给治疗金黄色葡萄球菌造成了严重的问题。金黄色葡萄球菌感染开发有效对抗这些致命MRSA菌株的药物的关键障碍之一是缺乏新的抗生素靶点。解决停滞核糖体的转译途径是药物开发的潜在目标，因为它存在于所有细菌中，并且是MRSA在培养物中生长所必需的。本项目的长期目标是确定特异性的反式翻译抑制剂。金黄色葡萄球菌，可用作药物开发的先导化合物。该提案的总体目标是验证反式翻译作为S中可行的抗生素靶标。金黄色葡萄球菌和开发一个初级检测兼容的高通量筛选（HTS）和二级检测，可用于识别抑制剂的反式翻译在S。金黄色。提出的研究的基本原理是，MRSA中的反式翻译作为药物靶标的验证和HTS测定的开发将导致快速鉴定新的抗生素，这将大大改善MRSA的治疗。该建议的具体目的是证实tmRNA和SmpB对于S.金黄色葡萄球菌的生长在培养和感染过程中，开发初级和二级筛选测定，以确定抑制剂的反式翻译在S。金黄色葡萄球菌，进行tmRNA-SmpB抑制剂的中试筛选，并使用已建立的离体和体内测定来表征筛选的命中。消耗菌株和小分子抑制剂将用于在培养物生长期间和感染期间去除反式翻译活性。这些实验结果将为理解S.金黄色葡萄球菌以及确定该途径是否可以用于药物开发。已建立的HTS试验和tmRNA-SmpB活性试验将适用于S.金黄色组分。这些研究的结果将允许未来的HTS和候选药物的高水平优化。通过靶向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疗法联合使用的新化合物。