Thiazolyl peptide analogs that provide insight into antibiotic targeting

噻唑基肽类似物可深入了解抗生素靶向

• 批准号: 8264573
• 负责人: Travis Scott Young
• 金额: $ 3.37万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8264573
• 关键词: Anabolism; Antibiotics; Bacteria; Cyclic Peptides; Data; Development; Diversity Library; Drug resistance; Gene Cluster; Gene Library; Genus staphylococcus; Goals; Gram-Positive Bacteria; Gray unit of radiation dose; Growth; Left; Libraries; Methicillin Resistance; Methods; Organism; Peptides; Plasmodium falciparum; Post-Translational Protein Processing; Production; Protein Biosynthesis; Public Health; Rattus; Research; Resistance; Streptomyces; Streptomyces lividans; Structure; Structure-Activity Relationship; Therapeutic; Wit; Work; analog; bacterial resistance; combat; genetic manipulation; insight; member; novel; novel therapeutics; pathogen; pathogenic bacteria; peptide analog; public health relevance; scaffold

DESCRIPTION (provided by applicant): Thiazolyl peptides possess novel scaffolds and proven activity against dangerous drug resistant pathogens such as methicillin-resistant Staphylococcus auerus (MRSA) and Plasmodium falciparum that make them ideal candidates for antibiotic development. Despite their potency and the pressing need for new antibiotics to combat emerging resistance, thiazolyl peptides have eluded development as therapeutics due to insufficient methods for their diversification. This deficiency has also hindered the study of structure-activity relationships to define how their novel scaffolds direct the antibiotic mode of action. The recent discovery that these molecules arise from ribosomal precursor peptides which undergo extensive post-translational modifications opens the door to rapid diversification through genetic manipulation. Towards this end, this research aims to create a novel thiazolyl peptide expression platform to enhance the understanding of how thiazolyl peptide scaffolds provide exquisite targeting of ribosomal protein synthesis in bacteria. Our specific hypothesis is that modified thiazolyl peptide ring structures will have alternate or multiple targets that inhibit the growth of gram positive bacteria. Preliminary data indicates the thiazolyl peptide biosynthetic machinery should readily accept modest changes in ring size. We intend to accomplish these goals through the following specific aims: 1. Identify the gene cluster responsible for the thiazolyl peptide GE37468 production in Streptomyces sp. ATCC 55365 and transfer it to the genetically tractable Streptomyces lividans to create a stable thiazolyl peptide expression platform. This will establish the first gene cluster from a publically accessible organism which produces a thiazolyl peptide with a 29-member ring and give insights into the biosynthesis of the unique GE37468 structure. 2. Create rational and high-diversity libraries of GE37468 analogs with altered ring sizes using the novel S. lividans expression platform. 3. Evaluate the mode of antibiotic activity for GE37468 analogs in a high through-put manner to create a better understanding of how structure provides targeting. The research proposed herein will take an unprecedented leap in the ability to create, understand, and screen libraries of cyclic peptides. This work has the potential to create thiazolyl peptides with novel spectrums of activity through the construction of high diversity genetic libraries. This is vital to the long-term goal of creating thiazolyl peptide therapeutics that can combat emerging resistance in pathogenic bacteria. PUBLIC HEALTH RELEVANCE: This project is relevant to public health because it proposes to develop greater understanding of how thiazolyl peptides inhibit protein synthesis in pathogenic bacteria. This understanding is vital to the development of new therapeutics to combat emerging bacterial resistance.

描述（由申请人提供）：噻唑基肽具有新型支架和经证实的对危险的耐药病原体如耐甲氧西林金黄色葡萄球菌（MRSA）和恶性疟原虫的活性，使其成为抗生素开发的理想候选物。尽管它们的效力和对新抗生素的迫切需求以对抗新出现的耐药性，但噻唑基肽由于其多样化的方法不足而未能作为治疗剂开发。这种缺陷也阻碍了结构-活性关系的研究，以确定其新型支架如何指导抗生素的作用模式。最近发现，这些分子来自核糖体前体肽，经过广泛的翻译后修饰打开了大门，通过遗传操作快速多样化。 为此，本研究旨在创建一个新的噻唑基肽表达平台，以增强对噻唑基肽支架如何提供细菌中核糖体蛋白合成的精确靶向的理解。我们的具体假设是，经修饰的噻唑基肽环结构将具有抑制革兰氏阳性菌生长的替代或多个靶点。初步数据表明，噻唑基肽生物合成机制应该很容易接受环大小的适度变化。我们打算通过以下具体目标来实现这些目标：1.鉴定链霉菌ATCC 55365中负责噻唑基肽GE 37468产生的基因簇，并将其转移到遗传上易处理的变铅青链霉菌中，以创建稳定的噻唑基肽表达平台。这将建立第一个基因簇，从一个生物可访问的生物体，产生一个噻唑肽与29个成员的环，并提供洞察生物合成的独特的GE 37468结构。2.使用新的S. lividans表达平台。3.以高通量方式评价GE 37468类似物的抗生素活性模式，以更好地了解结构如何提供靶向。 本文提出的研究将在创建，理解和筛选环肽库的能力方面实现前所未有的飞跃。这项工作有可能通过构建高多样性基因文库来创造具有新活性谱的噻唑基肽。这对于创造噻唑基肽治疗剂的长期目标至关重要，该治疗剂可以对抗病原菌中出现的耐药性。 公共卫生关系：该项目与公共卫生有关，因为它建议更好地了解噻唑基肽如何抑制病原菌中的蛋白质合成。这种理解对于开发新的治疗方法以对抗新出现的细菌耐药性至关重要。