MECHANISM OF DRUG-INDUCED DNA DEGRADATION

药物引起的 DNA 降解机制

• 批准号: 2177434
• 负责人: JOANNE STUBBE
• 金额: $ 29.64万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1997-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2177434
• 关键词: DNA damage; Escherichia coli; bleomycin; chemical structure function; conformation; drug interactions; free radicals; nuclear magnetic resonance spectroscopy; nucleic acid structure; stable isotope; stop flow technique

This application seeks continued support for mechanistic studies on the elucidation of the chemistry and biology of DNA cleavage by radical- generating drugs. Efforts in the past few years funded by this proposal have focused on designing new general methods to investigate the mechanism of these DNA cleavers and have been highly successful in unraveling the mechanism of the prototypical DNA cleaver bleomycin (BLM). The BLMs are a class of antitumor antibiotics isolated from S. verticillus that are used clinically against head and neck cancer and testicular cancer. Thus, a thorough understanding of its mechanism of action is of considerable relevance to the development of new chemotherapeutic agents. Work during the past funding period has laid the foundation for the comprehensive studies planned. In the next funding period, the following specific aims will be pursued: 1) The parameters responsible for the sequence, base and conformational specificity of BLM-mediated DNA cleavage will be examined. Methods employed will include isotope effect analysis and alternate nucleic acid structures such as DNA-RNA hybrids. 2) The structure and properties of single and double stranded lesions in DNA resulting from chemistry initiated by hydrogen atom abstraction at C-4' will be established using a variety of methods including NMR spectroscopy. 3) The mutagenicity and genotoxicity of these lesions in E. coli will be investigated. 4) Isotope effects will be used as a probe to examine the mechanism of double stranded cleavage of DNA by BLM. 5) Physical biochemical methods will be used to examine the structure of activated BLM and to reexamine the kinetics of its formation and destruction. 6) The mechanism of ene diyne antibiotics will be examined using isotope effect methods in conjunction with physical biochemical methods including stopped flow visible spectroscopy. These studies are designed to expand the detailed mechanistic insight that has been achieved through this research project into the broader issues of molecular recognition and biological effects of these important chemotherapeutic agents.

该应用寻求继续支持有关机械研究的 通过自由基阐明DNA裂解的化学和生物学 产生药物。 在过去的几年中，该提案资助 专注于设计新的一般方法来研究 这些DNA裂动物的机制，并且在 阐明原型DNA切动物博来霉素（BLM）的机制。 BLM是从S.分离的一类抗肿瘤抗生素。 在临床上针对头颈癌和 睾丸癌。 因此，对其机制的彻底理解 行动与新的发展相关 化学治疗剂。 在过去的资金期间工作已经奠定 综合研究的基础计划。 在下一个资金期间，将追求以下具体目标： 1）负责序列，碱基和构象的参数 将检查BLM介导的DNA裂解的特异性。 方法 使用的将包括同位素效应分析和交替的核酸 诸如DNA-RNA杂种之类的结构。 2）的结构和特性 由化学导致的DNA中的单链和双链病变 将使用C-4的氢原子抽象引发 包括NMR光谱在内的多种方法。 3）诱变性和 将研究大肠杆菌中这些病变的遗传毒性。 4） 同位素效应将用作检查的探针来检查 BLM对DNA的双链裂解。 5）物理生化方法 将用于检查激活的BLM的结构并重新检查 其形成和破坏的动力学。 6）ENE的机制 将使用同位素效应方法检查DIYNE抗生素 与物理生化方法（包括停止流量）的结合 可见光谱。 这些研究旨在扩大详细的机械洞察力 这是通过该研究项目实现的 这些重要的分子识别和生物学作用的问题 化学治疗剂。