Design and Characterization of DNA Interactive Agents

DNA 交互代理的设计和表征

• 批准号: 7922956
• 负责人: Jennifer S. Brodbelt
• 金额: $ 10万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7922956
• 关键词: Adverse effects; Alkylating Agents; Anthracyclines; Antineoplastic Agents; Arts; Binding; Binding Sites; Biochemical; Biological; Biological Assay; Chemicals; Clinical; Clinical Trials; Companions; Complex; DNA; DNA Adducts; DNA Binding; DNA Damage; DNA Fingerprinting; DNA Interstrand Crosslinking; DNA Repair; DNA Sequence; DNA Structure; DNA analysis; DNA lesion; DNA-Protein Interaction; Development; Diagnostic; Dissociation; Drug Binding Site; Enzymes; Evaluation; Ficusin; Gel; Goals; Histones; Ions; Ligase; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Metabolic; Methods; Molecular; Outcome; Pharmaceutical Preparations; Prodrugs; Proteins; Psoralens; Quinones; RNA chemical synthesis; Reaction; Recording of previous events; Research; Screening procedure; Site; Solutions; Spectrometry, Mass, Electrospray Ionization; Structure; Techniques; Transcription Process; adduct; analog; analytical method; anticancer activity; base; cancer cell; crosslink; cytotoxicity; design; diaziquone; drug candidate; drug mechanism; helicase; innovation; insight; interest; mass spectrometer; novel; novel strategies; pointed protein; public health relevance; tool; transcription factor

DESCRIPTION (provided by applicant): Bioreductive aziridinylbenzoquinones, one of the most potent classes of chemotherapeutic drugs, are alkylating agents that bind covalently to DNA. It is recognized that the basis for the activity of many anti-cancer agents is related to their ability to modify the structure of DNA, thus inhibiting DNA or RNA synthesis during DNA repair, replication, or transcription processes. The biochemical mechanisms of aziridinylbenzoquinones continue to be unraveled even as the search for more potent, more selective analogues with fewer toxic side effects moves forward. The clinical value of aziridinylbenzoquinones has promoted ongoing interest in developing new analogues, and this continuing interest has spurred the need for sensitive, versatile analytical methods that can be used to determine the structures of the resulting DNA adducts and DNA crosslinks, to evaluate the DNA sequence/site selectivities and reactivities of these alkylating agents, and to better understand the ways that the resulting DNA adducts interact with relevant proteins in a way that leads to anticancer activities. This proposal will describe the development and application of several innovative mass spectrometric strategies for the structural characterization of DNA adducts and chemical probe strategies for targeting DNA adducts and protein/DNA interactions. The overall goal is to develop new photodissociation techniques in conjunction with electrospray ionization mass spectrometry, as well as chemical probe methods based on selective reactivities and derivatization strategies, to create a tool-kit of mass spectrometric methods for characterizing new DNA alkylating agents. Our innovative technological approaches and the resulting outcomes will provide insights into site/sequence selectivity of alkylating agents, offer rapid and sensitive structural characterization of DNA adducts, and offer a new approach for mapping DNA/protein interactions. An array of bioreductive aziridinylbenzoquinones will be synthesized, including ones in which the alkyl substituents are modified to alter the reduction potentials and modulate their cytotoxicities, with the aim of increasing the selectivity of the quinones and constructing structure/reactivity relationships based on the ESI-MS (electrospray ionization mass spectrometry) methods and companion cytoxicity assays. PUBLIC HEALTH RELEVANCE: The activities of many anticancer agents are dependent on their ability to bind DNA, either via formation of interstrand cross-links or formation of monoadducts which involve binding to a single DNA site. This research enables the design of new anticancer aziridinylbenzoquinones and advances the analysis of DNA-drug adducts by innovative mass spectrometric strategies.

描述（由申请人提供）：生物还原性氮丙啶基苯醌是一类最有效的化疗药物，是与DNA共价结合的烷化剂。人们认识到，许多抗癌剂的活性基础与它们修饰DNA结构的能力有关，从而在DNA修复、复制或转录过程中抑制DNA或RNA合成。氮丙啶基苯醌的生化机制继续被解开，即使寻找更有效的，更有选择性的类似物，更少的毒副作用向前迈进。氮丙啶基苯醌的临床价值促进了对开发新的类似物的持续兴趣，并且这种持续的兴趣刺激了对灵敏的、通用的分析方法的需求，所述分析方法可用于确定所得DNA加合物和DNA交联的结构，以评估这些烷化剂的DNA序列/位点选择性和反应性，并更好地了解所产生的DNA加合物与相关蛋白质相互作用的方式，从而导致抗癌活性。该提案将描述几种创新的质谱策略的发展和应用，用于DNA加合物的结构表征和针对DNA加合物和蛋白质/DNA相互作用的化学探针策略。总体目标是开发新的光解离技术与电喷雾电离质谱，以及化学探针方法的基础上选择性反应性和衍生化策略，以创建一个工具包的质谱方法表征新的DNA烷化剂。我们的创新技术方法和由此产生的结果将提供对烷化剂的位点/序列选择性的深入了解，提供DNA加合物的快速和灵敏的结构表征，并提供绘制DNA/蛋白质相互作用的新方法。将合成一系列生物还原性氮丙啶基苯醌，包括其中烷基取代基被修饰以改变还原电位并调节其细胞毒性的那些，目的是增加醌的选择性并基于ESI-MS（电喷雾电离质谱）方法和伴随细胞毒性测定构建结构/反应性关系。公共卫生相关性：许多抗癌剂的活性取决于它们结合DNA的能力，通过形成链间交联或形成涉及结合单个DNA位点的单加合物。这项研究使新的抗癌aziridinylbenzoquinones的设计和先进的分析DNA药物加合物的创新质谱策略。