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：
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
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/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

项目摘要

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）：艾尔德素天然产物的衍生物酰基化是癌症治疗的有前途的药物。它们是肿瘤细胞的高治疗指数和选择性的知名度，并且正在进行临床试验。酰基含量是烷基化剂，但是DNA和蛋白质成型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机械范式，以核算酰基化的选择性毒性，这将导致旨在特定于特定于特定于特定特定性调节酰基饱和毒性的组合和量身定制的疗法的发展。该应用的目标是建立从酰基饱和生物活化到生物分子靶标修饰和细胞毒性的联系。具体的目的是（1）表征通过酰基化的体外DNA烷基化的体外模式，并区分了直接修饰DNA与生物活化偶联反应的加合物；（2）表征了酰基化抑制细胞氧化还原蛋白的机制；（3）确定生物活化还原酶对细胞中酰基饱和烷基化反应和毒性的影响。 AIM 1的研究方法涉及化学活化的分子探针的合成，该探针将用于鉴定通过生物活化的酰基成分鉴定DNA烷基化的产物。我们将使用化学表征方法来识别酰基饱和DNA加合物的结构。在AIM 2中，我们将使用酶抑制研究，并结合基于质谱的方法来绘制结构绘制蛋白质共价修饰的方法。在AIM 3中，我们将使用过表达的细胞和稳定的肿瘤细胞系来测试加合物形成在细胞复杂环境中的影响。酰基化是一类新的药物，它们通过与细胞中的生物分子并启动导致细胞死亡的一系列生物事件的序列杀死癌细胞。控制这一过程的最初化学转化以及它们在细胞中如何差异，这些细胞对酰基化的敏感性与未知的细胞敏感。选择性靶向肿瘤细胞的能力是癌症治疗中的一个核心问题，因此了解这种机制对于设计更安全的癌症治疗策略很重要。