Mass spectrometry Studies on Radical Reactions of DNA

DNA自由基反应的质谱研究

• 批准号: 7350749
• 负责人: HILKKA Inkeri KENTTAMAA
• 金额: $ 4.09万
• 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 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7350749
• 关键词: Acoustics; Address; Antineoplastic Agents; Antiviral Agents; Area; Behavior; Binding; Biological; Cell Death; Cells; Characteristics; Charge; Cleaved cell; Collection; Complement; Complex; Condition; DNA; Data; Deoxyribose; Depth; Depurination; Development; Drug Binding Site; Drug Design; Funding; Galium; Gases; Generations; Goals; Guidelines; Hydrogen; Individual; Ink; Kinetics; Knowledge; Lasers; Lead; Learning; Light; Lung; Malignant Neoplasms; Malignant neoplasm of ovary; Mass Spectrum Analysis; Methods; Modeling; Modification; Molecular; Mono-S; Oligonucleotide Probes; Oligonucleotides; Organic Chemistry; Peptides; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Phase; Positioning Attribute; Process; Prodrugs; Proteins; Range; Reaction; Research; Scientist; Site; Solutions; Specialist; Structure; System; Testing; Thinking; Work; abstracting; base; cancer cell; cellular targeting; chemical property; cytotoxicity; design; insight; instrumentation; interest; molecular orbital; novel; professor; research study; sugar

The action of many non-hydrolytic DNA cleaving agents is believed to be based on reactive phenyl radicals that abstract hydrogen atoms from deoxyribose in DNA. Various issues relating to this process, including the drug's binding site on DNA, the mechanism of radical 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 and antiviral drugs. However, the key part of the process, the reaction of the radical intermediate with DNA, remains unexplored. Although information on the factors that control this reaction is critically needed for drug design, nearly nothing is known about the radical intermediates formed from drugs because of severe experimental difficulties in studying such highly reactive species. Our goal is to learn how to control the reactivity and intrinsic selectivity of a radical warhead by structural changes. We have developed a novel experimental approach based on mass spectrometry to study biologically relevant radical reactions. The method involves attaching a charged ¿group to a radical of interest for manipulation in an FT-ICR m_;ss spectrometer wherein small neutral biomolecules are introduced by laser-induced acoustic desorption. This proposal presents a continuation of a four-year project that involved 1) purchase and set-up of the necessary instrumentation, 2) demonstration of the feasibility of the approach, 3) examination of known systems to show that the method yields data relevant to neutral radicals and solution conditions, and 4) collection of kinetic reactivity data to provide several novel structure/reactivity relationships for advancement of the design of better non-hydrolytic DNA cleavers. The most important areas of the currently proposed work include (1) d6velopment of structure/reactivity relationships for more complex systems, (2) exploration of the selectivity of different radicals toward different sites in oligonucleotides, (3) examination of the propensity of the radicals to attack peptides to model protein damage, and (4) based on the insights gained in the above studies, development of a reactivity paradigm that provides general predictive power. (5) This paradigm will be used to design target-specific radical warheads and prodrugs. Professors Rick Borch (Medicinal Chemistry and Molecular Pharmacology, Purdue), a specialist in cancer drug design and mechanisms, I ¿ and Mark Lipton (Organic Chemistry, Purdue), an expert in synthesis of biradical producing prodrugs, will guide t the research and help in findin.q the best way to incorporate knowled,qe into the desi,qn and testing of novel drugs.]

许多非水解性DNA裂解剂的作用被认为是基于活性苯基，这些苯基 从DNA中的脱氧核糖中提取氢原子。与这一过程有关的各种问题，包括药物的 DNA上的结合部位，自由基的形成机制，它与不同DNA部位的可及性，以及DNA的命运 由此产生的DNA自由基，正在被科学家们解决，以推进更好的抗肿瘤和 抗病毒药物。然而，这一过程的关键部分，即自由基中间体与DNA的反应，仍然存在 未被开发的。尽管药物设计非常需要关于控制这种反应的因素的信息， 由于严格的实验，人们对药物形成的自由基中间体几乎一无所知 研究如此高活性物种的困难。我们的目标是学习如何控制反应性和内在的 通过结构变化实现激进弹头的选择性。我们开发了一种新的实验方法，基于 关于研究生物相关自由基反应的质谱学。该方法包括将带电的 在FT-ICR m_；ss光谱仪中操作的感兴趣的基团，其中小的中性生物分子 都是通过激光诱导的声解吸引入的。这项提案是一个为期四年的项目的延续。 这包括1)购买和设置必要的仪器，2)论证 方法，3)对已知系统的检查，以表明该方法产生与中性自由基和 溶解条件，以及4)收集动力学反应性数据以提供几种新结构/反应性 设计更好的非水解型dna切割器的关系。最重要的领域 目前提议工作包括：(1)发展更复杂的结构/反应关系 系统，(2)不同基团对寡核苷酸不同位置的选择性的探索，(3) 检查自由基攻击多肽以模拟蛋白质损伤的倾向，以及(4)基于 在上述研究中获得的见解，开发提供一般预测能力的反应性范式。 (5)这一范例将被用于设计特定目标的激进型弹头和前体药物。里克·博尔奇教授 (药物化学和分子药理学，普渡大学)，抗癌药物设计和机制方面的专家，I？ 和马克·利普顿(普渡大学有机化学)，一位合成产生前体药物的双自由基的专家，将指导 研究和帮助找到将Knowleed、QE纳入新药设计、QN和测试的最佳方式。]