Design and Characterization of DNA Interactive Agents

描述（由申请人提供）：生物还原性aziridinylbenzoquinones是最有效的一类化疗药物之一，是与DNA共价结合的烷基化剂。人们认识到，许多抗癌药物的活性基础与其修饰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的合成。即使在寻找更有效、更有选择性、毒副作用更小的类似物的过程中，aziridinylbenzoquinones的生化机制仍在继续被揭示。aziridinylbenzoquinones的临床价值促进了对开发新的类似物的持续兴趣，这种持续的兴趣刺激了对灵敏，通用的分析方法的需求，这些分析方法可用于确定所产生的DNA加合物和DNA交联的结构，以评估这些烷基化剂的DNA序列/位点选择性和反应性。并更好地了解产生的DNA加合物与相关蛋白质相互作用的方式，从而产生抗癌活性。本提案将描述几种用于DNA加合物结构表征的创新质谱策略和用于靶向DNA加合物和蛋白质/DNA相互作用的化学探针策略的发展和应用。总体目标是开发新的光解离技术，结合电喷雾电离质谱法，以及基于选择性反应性和衍生化策略的化学探针方法，创建一个用于表征新型DNA烷基化剂的质谱方法工具包。我们的创新技术方法和结果将为烷基化剂的位点/序列选择性提供见解，提供DNA加合物的快速和敏感的结构表征，并为绘制DNA/蛋白质相互作用提供新的方法。本文将合成一系列具有生物还原性的氮基苯基醌类化合物，包括通过修饰烷基取代基来改变还原电位和调节其细胞毒性的化合物，目的是提高醌类化合物的选择性，并基于ESI-MS（电喷雾电离质谱）方法和相应的细胞毒性测定建立结构/反应性关系。公共卫生相关性：许多抗癌药物的活性取决于它们结合DNA的能力，要么通过形成链间交联，要么通过形成单加合物与单个DNA位点结合。本研究为设计新型抗癌亚氮基苯醌提供了可能，并通过创新的质谱分析策略推进了dna -药物加合物的分析。

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.