BIOSYNTHESIS OF ANTITUBERCULAR NONRIBOSOMAL PEPTIDES

抗结核非核糖体肽的生物合成

• 批准号: 6746585
• 负责人: T. MARK ZABRISKIE
• 金额: $ 24.03万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6746585
• 关键词: aminoacid biosynthesis; antitubercular agents; biotechnology; drug discovery /isolation; enzyme activity; functional /structural genomics; gene expression; ligase; nucleic acid sequence; proteomics

DESCRIPTION (provided by applicant): Tuberculosis (TB) kills more young and middle-aged adults worldwide than any other disease with the exception of AIDS. Furthermore, TB is the leading cause of morbidity and mortality among the population living with HIV/AIDS. While TB is generally perceived as a disease afflicting the poorest countries, the rapid increase in cases of multidrug-resistant tuberculosis (MDR-TB) and its spread to industrialized nations makes the disease a global threat. As primary drugs are rendered ineffective, second-line agents that are more expensive, usually less effective and often more toxic must be used. Hence, the need for new anti-TB drugs has never been greater, yet it has been over 30 years since the introduction of the last novel antitubercular agent. Advances to improve potency, decrease toxicity, and shorten the duration of therapy would have an enormous impact on world health and healthcare costs. This application describes an approach to access, utilize and manipulate peptide biosynthesis genes that direct precursor construction and post-assembly tailoring to create novel antitubercular peptides. The targets are analogs of viomycin, a peptide antibiotic produced by Streptomyces vinaceus and a second-line antitubercular agent that is often effective against MDR-TB. Viomycin possesses rare and unique amino acids that are crucial for biological activity but are not available for chemical synthesis. The proposal describes studies on the biochemical transformations leading to these novel amino acids, including reactions proposed to occur on peptidyl carrier (PCP) protein-bound species. Experiments are proposed to generate new peptides through targeted gene disruption and directed biosynthesis with precursor analogs added exogenously or formed in situ. Novel compounds will also be formed by controlling enzymes that decorate and fully activate the core peptide. Successfully harnessing the genes and deciphering the biochemical mechanisms involved in viomycin precursor assembly and tailoring will yield valuable information that will be translated into novel molecular tools for generating new antitubercular agents. The knowledge and methods that arise from these studies will be directly applicable to expanding the chemical diversity in other families of bioactive peptides.

描述（由申请人提供）：全世界死于结核病（TB）的青壮年人数比除艾滋病之外的任何其他疾病都要多。此外，结核病是艾滋病毒/艾滋病感染者发病和死亡的主要原因。虽然结核病通常被认为是一种困扰最贫穷国家的疾病，但耐多药结核病 (MDR-TB) 病例的迅速增加及其向工业化国家的传播使该疾病成为全球威胁。由于主要药物变得无效，必须使用更昂贵、通常效果较差且毒性更大的二线药物。因此，对新型抗结核药物的需求从未如此之大，但距离上一种新型抗结核药物的推出已经过去了 30 多年。提高效力、降低毒性和缩短治疗持续时间的进步将对世界健康和医疗保健成本产生巨大影响。该申请描述了一种获取、利用和操纵肽生物合成基因的方法，该基因指导前体构建和组装后剪裁以创建新型抗结核肽。目标是紫霉素的类似物，紫霉素是一种由酒红色链霉菌产生的肽抗生素，也是一种二线抗结核药物，通常对耐多药结核病有效。紫霉素拥有稀有且独特的氨基酸，这些氨基酸对于生物活性至关重要，但无法用于化学合成。该提案描述了对产生这些新型氨基酸的生化转化的研究，包括提议在肽基载体（PCP）蛋白质结合物种上发生的反应。建议进行实验，通过靶向基因破坏和外源添加或原位形成的前体类似物进行定向生物合成来产生新的肽。通过控制装饰和完全激活核心肽的酶也将形成新的化合物。成功利用基因并破译涉及紫霉素前体组装和剪裁的生化机制将产生有价值的信息，这些信息将转化为用于产生新抗结核药物的新型分子工具。这些研究产生的知识和方法将直接适用于扩大其他生物活性肽家族的化学多样性。