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）：对现有抗生素具有抗药性的金黄色葡萄球菌菌株的持续传播给治疗金黄色葡萄球菌感染造成了严重的问题。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。用于解决停滞核糖体问题的反式翻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。该项目的长期目标是确定金黄色葡萄球菌中可用作药物开发先导化合物的特异性反式翻译抑制剂。该提案的总体目标是验证反式翻译作为金黄色葡萄球菌中可行的抗生素靶点，并开发与高通量筛选（HTS）兼容的初级测定和可用于鉴定金黄色葡萄球菌中反式翻译抑制剂的二级测定。拟议研究的基本原理是，验证 MRSA 中的反式翻译作为药物靶标以及开发 HTS 检测方法将导致快速鉴定新抗生素，从而显着改善 MRSA 的治疗。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的生长至关重要，开发初级和二级筛选测定法来鉴定金黄色葡萄球菌中的反式翻译抑制剂，对 tmRNA-SmpB 抑制剂进行初步筛选，并使用已建立的离体和体内测定法来表征筛选的命中结果。耗竭菌株和小分子抑制剂将用于消除培养物生长期间和感染期间的转译活性。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以及确定该途径是否可以作为药物开发的目标提供基础。已建立的 HTS 测定和 tmRNA-SmpB 活性测定将适用于金黄色葡萄球菌成分。这些研究的结果将为未来的 HTS 和候选药物的高水平优化提供支持。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 疗法联合使用的新化合物。