Covalent Modification of DNA and Protein by Bioactivated Antitumor Acylfulvenes

生物活性抗肿瘤酰基富烯对 DNA 和蛋白质的共价修饰

• 批准号: 7259793
• 负责人: SHANA J STURLA
• 金额: $ 24.29万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-23 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7259793
• 关键词: Abbreviations; Accounting; Address; Affect; Alkylating Agents; Alkylation; Antineoplastic Agents; Apoptosis; Binding; Biochemical Process; Biological; Biological Factors; Biological Process; Buffers; Cell Death; Cells; Chemical Structure; Chemicals; Class; Clinical; Clinical Research; Clinical Trials; Combined Modality Therapy; Complex; Coupled; DNA; DNA Adducts; DNA Alkylation; DNA Damage; DNA Modification Process; DNA Repair; DNA biosynthesis; DNA chemical synthesis; Data; Development; Disruption; Dose-Limiting; Engineering; Environment; Enzyme Inhibition; Enzymes; Event; Future; Genetic Transcription; Goals; In Vitro; Investigation; Knowledge; Lead; Link; Maps; Mass Spectrum Analysis; Mediating; Metabolic; Metabolism; Methodology; Methods; Modeling; Modification; Molecular Probes; Nature; Nucleosides; Oxidation-Reduction; Oxidoreductase; Pathway interactions; Pattern; Pharmaceutical Preparations; Principal Investigator; Process; Protein Overexpression; Proteins; Publishing; Reaction; Reactive Oxygen Species; Reagent; Reporting; Research; Research Design; Resistance; Role; Signal Pathway; Signal Transduction; Site; Site-Directed Mutagenesis; Structure; System; TP53 gene; Testing; Therapeutic Index; Thioredoxin; Toxic effect; Tumor Cell Line; Work; acylfulvene; adduct; analog; analytical tool; base; cancer cell; cancer therapy; cellular engineering; cellular targeting; chemical carcinogenesis; chemical synthesis; chemotherapeutic agent; cytotoxicity; design; diene; drug sensitivity; enzyme activity; illudin M; improved; innovation; irofulven; killings; neoplastic cell; nucleobase; small molecule; therapy design; thioredoxin reductase; tumor

DESCRIPTION (provided by applicant): Acylfulvenes, derivatives of illudin natural products, are a promising class of drugs for cancer treatment. They are notable for high therapeutic indices and selectivities for tumor cells, and clinical trials are underway. Acylfulvenes are alkylating agents, but the detailed chemical mechanisms of DNA and protein-adduct formation, and how this relates to tumor-cell selectivity is not known. The long-term goals of this project are to define a mechanistic paradigm accounting for the selective toxicities of acylfulvenes that will lead to the development of combined-agent and tailored therapies designed to site-specifically tune acylfulvene toxicity. The objectives of this application are to establish a link from acylfulvene bioactivation to biomolecular target modification and cytotoxicity. The specific aims are to (1) Characterize in vitro patterns of DNA alkylation by acylfulvenes and distinguish adducts that result from direct modification of DNA from those of bioactivation-coupled reactions; (2) Characterize the mechanism of inhibition of cellular redox proteins by acylfulvenes; (3) Determine the influence of a bioactivating reductase on acylfulvene alkylation reactions and toxicity in cells. The research approach for aim 1 involves the synthesis of chemically activated molecular probes that will be used to identify products of DNA alkylation by bioactivated acylfulvenes. We will use chemical characterization methods to identify the structures of acylfulvene DNA adducts. In aim 2, we will use enzyme-inhibition studies combined with a mass-spectrometry-based approach to structurally map protein covalent modification. In aim 3, we will use reductase-overexpressing cells and stable tumor cell lines to test the impact of adduct formation in the complex environment of the cell. Acylfulvenes are a new class of drugs that kill cancer cells by becoming covalently linked to biomolecules in the cell and initiating a sequence of biological events that result in cell death. The initial chemical transformations that govern this process and how they differ in cells that are sensitive to acylfulvenes versus those that are not are unknown. The ability to selectively target tumor cells is a central problem in cancer therapy and consequently understanding this mechanism is important for designing safer cancer therapy strategies.

描述（由申请人提供）：酰基富烯，隐花素天然产物的衍生物，是一类有前途的癌症治疗药物。它们以高治疗指数和对肿瘤细胞的选择性而闻名，临床试验正在进行中。酰基富烯是烷化剂，但DNA和蛋白质加合物形成的详细化学机制，以及这与肿瘤细胞选择性的关系尚不清楚。该项目的长期目标是定义一种机制范式，解释酰基富烯的选择性毒性，这将导致开发联合药物和定制的治疗方法，旨在对酰基富烯毒性进行位点特异性调节。本申请的目的是建立从酰基富烯生物活化到生物分子靶向修饰和细胞毒性的联系。具体目标是：（1）表征酰基富烯对DNA烷基化的体外模式，并区分直接修饰DNA的加合物与生物活化偶联反应的加合物;（2）表征酰基富烯抑制细胞氧化还原蛋白的机制;（3）确定生物活化还原酶对酰基富烯烷基化反应的影响和细胞毒性。目标1的研究方法涉及化学活化的分子探针的合成，这些探针将用于鉴定生物活化的酰基富烯的DNA烷基化产物。我们将使用化学表征方法来鉴定酰基富烯DNA加合物的结构。在目标2中，我们将使用酶抑制研究结合质谱法为基础的方法来结构映射蛋白质共价修饰。在目标3中，我们将使用过表达还原酶的细胞和稳定的肿瘤细胞系来测试在细胞的复杂环境中加合物形成的影响。酰基富烯是一类新型药物，通过与细胞中的生物分子共价连接并引发导致细胞死亡的一系列生物事件来杀死癌细胞。控制这一过程的初始化学转化以及它们在对酰基富烯敏感的细胞与对酰基富烯不敏感的细胞中的差异尚不清楚。选择性靶向肿瘤细胞的能力是癌症治疗中的核心问题，因此了解这种机制对于设计更安全的癌症治疗策略非常重要。