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

基本信息

批准号：
7611449
负责人：
SHANA J STURLA
金额：
$ 4.5万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
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/7611449
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 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 method 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 tool tumor

项目摘要

Project Summary: Acylfulvenes, derivatives of illudin natural products, are a promising class of drugs for cancer treatment. They are notable for high therapeutic indices and selectivitiesfor 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 selectivetoxicities of acylfulvenes that will lead to the development of combined-agent and tailored therapies designed to site-specifically tune acylfulvene toxicity. The objectivesof 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 bioactivatedacylfulvenes. 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-overexpressingcells and stable tumor cell lines to test the impact of adduct formation in the complex environment of the cell. Relevance: 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 selectivelytarget tumor cells is a central problem in cancer therapy and consequently understanding this mechanism is important for designing safer cancer therapy strategies.

项目概要：酰基富烯是伞菌素天然产物的衍生物，是一类很有前景的癌症治疗药物。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 和蛋白质加合物的详细化学机制形成，以及这与肿瘤细胞选择性有何关系尚不清楚。该项目的长期目标是定义一个解释酰基富烯选择性毒性的机械范式，这将导致开发旨在特异性调节酰基富烯毒性的组合药物和定制疗法。本申请的目的是建立从酰富烯生物活化到生物分子靶标的联系修饰和细胞毒性。具体目标是 (1) 通过以下方式表征 DNA 烷基化的体外模式：酰基富烯，并将 DNA 直接修饰产生的加合物与生物活化的加合物区分开来耦合反应； (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和细胞毒性的影响。目标 1 的研究方法涉及化学活化分子探针的合成，这些探针将被用于鉴定生物活性酰基富烯的 DNA 烷基化产物。我们将使用化学品鉴定酰基富烯 DNA 加合物结构的表征方法。在目标 2 中，我们将使用酶抑制研究与基于质谱的方法相结合来绘制蛋白质结构图共价修饰。在目标3中，我们将使用还原酶过表达细胞和稳定的肿瘤细胞系来测试细胞复杂环境中加合物形成的影响。关联：酰富烯是一类新型药物，通过与生物分子共价连接来杀死癌细胞细胞并启动一系列导致细胞死亡的生物事件。最初的化学物质控制这一过程的转化以及它们在对酰基富烯敏感的细胞和对酰基富烯敏感的细胞中有何不同那些不是的则未知。选择性靶向肿瘤细胞的能力是癌症的核心问题治疗并因此了解这一机制对于设计更安全的癌症治疗非常重要策略。