Mass Spectrometry Studies on Radical Reactions of DNA

DNA 自由基反应的质谱研究

• 批准号: 7651536
• 负责人: HILKKA Inkeri KENTTAMAA
• 金额: $ 2.45万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7651536
• 关键词: Acoustics; Address; Aging; Alzheimer' s Disease; Amino Acids; Antineoplastic Agents; Binding Sites; Biological; Biopolymers; Cell Death; Charge; Chemistry; Classification; Cleaved cell; Clinical; Complex; DNA; DNA-Protein Interaction; Deoxyribose; Development; Disease; Drug Design; Family; Funding; Gases; Generations; Goals; Hydrogen; Ions; Knowledge; Lasers; Lung; Malignant Neoplasms; Malignant neoplasm of ovary; Mass Spectrum Analysis; Methodology; Methods; Molecular; Mono-S; Nerve Degeneration; Nucleic Acids; Oral cavity; Peptides; Pharmaceutical Preparations; Phase; Photochemistry; Positioning Attribute; Process; Prodrugs; Proteins; Radiation therapy; Reaction; Research; Role; Scientist; Site; Solutions; Structure; Superhelical DNA; Surface; System; Technology; Testing; Thinking; Toxic effect; United States National Institutes of Health; Water; abstracting; anticancer research; antitumor drug; base; cancer site; chemical property; clinical application; computer studies; cytotoxicity; design; drug development; ds-DNA; gel electrophoresis; improved; instrument; interest; irradiation; mass spectrometer; novel; oxidative DNA damage; photolysis; professor; protein aminoacid sequence

DESCRIPTION (provided by applicant): The action of the most potent family of anticancer agents discovered thus far, the enediynes, is believed to be based on reactive ???-biradicals that abstract a hydrogen atom from each strand of double-stranded DNA. Unfortunately, the high toxicity of these agents hinders their clinical use. Various issues relating to their action, including their binding sites in DNA, the mechanism of biradical formation, its accessibility to various DNA sites, and the fate of the resulting DNA radicals, are being addressed by scientists to advance the rational design of better antitumor drugs. However, the key part of the process, the reaction of the biradical intermediate with DNA and other cellular components, remains almost entirely unexplored. Although information on the factors that control these reactions is critically needed for rational drug design, nearly nothing is known about the biradicals formed from drugs because of severe experimental difficulties in studying such highly reactive species. The goal of this research is to provide information on the role of these biradicals in cellular damage, and to advance approaches of controlling their reactivity and selectivity for drug development. A unique mass spectrometric method (developed with NIH support) for the study of biological radical reactions in the gas phase will be employed. The method involves attaching a charged group to a biradical of interest for manipulation in an FT-ICR mass spectrometer into which neutral biomolecules are introduced by laser-induced acoustic desorption. In the past, this method allowed the delineation of novel structure/reactivity relationships for reactions of mono- and biradicals with small DNA components, amino acids and small peptides. The proposed research will include 1) further studies on "tuning" of the reactivity and selectivity of biradicals, and inclusion of polyradicals, 2) examination of more complex biopolymers in order to advance a reactivity paradigm that provides predictive power for DNA and proteins, 3) development of methodology for the generation of biradicals in solution and testing of their DNA cleaving ability, and finally, 4) the mass spectrometry technology will be implemented in commercial instruments to make it accessible to other scientists, and the potential for peptide sequencing of radical reactions will be pursued. Furthermore, by improving the fundamental knowledge on oxidative DNA damage, this research advances the general understanding of aging and various diseases, including cancer. This research will drastically improve the fundamental knowledge on oxidative DNA and protein damage. Systematic examination of drug intermediate/DNA and protein interactions will enable the design of better synthetic antitumor drugs for the treatment of, for example, lung, mouth and ovarian cancers. Further, this research advances the general understanding of aging and various diseases, including cancer.

说明书(申请人提供)：到目前为止发现的最有效的抗癌药物家族，烯二炔类，被认为是基于反应性的？？-双自由基，从双链DNA的每条链中提取一个氢原子。不幸的是，这些药物的高毒性阻碍了它们的临床应用。与它们的作用有关的各种问题，包括它们在DNA中的结合部位，双自由基的形成机制，它与不同DNA位点的可及性，以及产生的DNA自由基的去向，都是科学家们正在研究的问题，以促进更好的抗肿瘤药物的合理设计。然而，这一过程的关键部分，即双自由基中间体与DNA和其他细胞成分的反应，几乎完全没有被探索过。尽管控制这些反应的因素的信息对于合理的药物设计是至关重要的，但由于研究这种高活性物种的严重实验困难，人们对药物形成的双自由基几乎一无所知。这项研究的目的是提供关于这些双自由基在细胞损伤中的作用的信息，并促进控制它们的反应性和药物开发的选择性的方法。将使用一种独特的质谱学方法(在NIH支持下开发)来研究气相中的生物自由基反应。该方法包括将带电基团连接到感兴趣的双自由基上，以便在FT-ICR质谱仪中进行操作，其中中性生物分子通过激光诱导的声解吸引入其中。在过去，这种方法可以描述单自由基和双自由基与小DNA组分、氨基酸和小肽反应的新的结构/反应关系。拟议的研究将包括1)进一步研究双自由基的反应性和选择性以及多自由基的包含性，2)检查更复杂的生物聚合物，以推进为DNA和蛋白质提供预测能力的反应性范例，3)开发在溶液中生成双自由基的方法并测试其DNA切割能力，最后，4)将在商业仪器中实施质谱学技术，使其他科学家能够使用它，并探索对自由基反应进行肽序列的可能性。此外，通过提高氧化DNA损伤的基础知识，这项研究促进了对衰老和包括癌症在内的各种疾病的总体理解。 这项研究将极大地提高对DNA氧化和蛋白质损伤的基础知识。对药物中间体/DNA和蛋白质相互作用的系统研究将能够设计出更好的合成抗肿瘤药物，用于治疗例如肺癌、口腔癌和卵巢癌。此外，这项研究还促进了对衰老和包括癌症在内的各种疾病的总体理解。