INTERACTIONS OF ANTITUMOR AGENTS WITH NUCLEIC ACIDS

抗肿瘤剂与核酸的相互作用

• 批准号: 3563742
• 负责人: David E Graves
• 金额: $ 4.75万
• 依托单位: UNIVERSITY OF MISSISSIPPI
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-03-01 至 1989-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3563742
• 关键词: DNA; affinity labeling; aminoacridines; antineoplastic antibiotics; antineoplastics; chemical binding; chemical structure function; chromatin; covalent bond; nuclear magnetic resonance spectroscopy; nucleic acid chemical synthesis; thermodynamics; ultraviolet spectrometry

This project will explore the molecular, thermodynamic, and biochemical properties associated with the interactions of the potent antitumor antibiotics m-AMSA and actinomycin D with nucleic acids. Specifically, these research efforts will be designed to explore the structural and functional properties of native and synthetic DNAs subsequent to drug binding utilizing both equilibrium and covalent modes of ligand interactions and correlate the relationships of drug structure with the biophysical parameters associated with complex formation. Experiments will be designed to extend the binding isotherms of m-AMSA to very low concentrations of bound drug, providing a more accurate description of the physical chemical nature of its chemotherapeutic effect. Thermodynamic, kinetic, and stoichiometric measurements will be obtained by spectroscopic, equilibrium dialysis, and phase partition techniques. The phase partition method will be particularly useful for obtaining accurate binding isotherms at very low concentrations of bound drug. The technique of photoaffinity labeling will be extended to the actinomycin D system. The capacity for covalent attachment will greatly aid in the study of target site specificities, cooperativity, and/or drug clustering phenomena. Through chemical modification of the parent actinomycin D with a photoreactive substituent (azido moiety), functional analogs of the parent molecule will be generated having equilibrium binding properties identical to those of the parent compound (in the absence of light) yet with the capacity for covalent attachment upon photolytic activation. Properties associated with the covalent nature of the actinomycin D - nucleic acid adduct will be examined in detail by a variety of biochemical and biophysical techniques. Restriction endonuclease activity will be used to monitor binding site specificities and the nucleic acid structural perturbations resulting from adduct formation. Similarly, the geometry and molecular orientation of the actinomycin D - DNA adduct will be determined at the oligonucleotide level by high field nuclear magnetic resonance spectroscopy. These studies on both m-AMSA and actinomycin D will increase the understanding of drug binding properties, the nature of the drug - DNA complex and/or adduct, and provide a powerful photoaffinity probe for further definition of the structural and functional properties of nucleic acids.

该项目将探索分子，热力学和生化 与有效抗肿瘤的相互作用相关的特性 抗生素M-AMSA和放线霉素D与核酸。 具体来说， 这些研究工作将旨在探索结构和 药物之后的天然和合成DNA的功能特性 利用配体的平衡和共价模式的结合 相互作用并将药物结构的关系与 与复杂形成相关的生物物理参数。 实验将旨在将M-AMSA的结合等温等温线扩展到 非常低浓度的结合药物，提供更准确的 其化学治疗的物理化学性质的描述 影响。 热力学，动力学和化学计量测量将是 通过光谱，平衡透析和相分解获得 技术。 相分区方法将特别有用 在非常低的结合处获得准确的结合等温线 药品。 光性标签的技术将扩展到放线霉素 D系统。 共价附件的能力将极大地帮助 研究目标场地特异性，合作和/或药物聚类的研究 现象。 通过化学修饰父母放线霉素D 光电反应性取代基（方唑部分），功能类似物的功能类似物 父分子将产生具有平衡结合特性 与父母化合物的相同（在没有光的情况下） 光解激活后具有共价附着的能力。 与放线霉素D-的共价性质相关的特性 核酸加合物将通过多种生化进行详细检查 和生物物理技术。 将使用限制性核酸内切酶活动 监测结合位点特异性和核酸结构 加合物形成产生的扰动。 同样，几何形状和 放线菌素D -DNA加合物的分子取向将确定 在高场核磁共振上，在寡核苷酸水平上 光谱法。 这些对M-AMSA和放线霉素D的研究将增加 了解药物结合特性，药物的性质 - DNA 复杂和/或加合物，并为 核的结构和功能特性的进一步定义 酸。