INTERACTIONS OF ANTITUMOR AGENTS WITH NUCLEIC ACIDS

抗肿瘤剂与核酸的相互作用

• 批准号: 3457873
• 负责人: David E Graves
• 金额: $ 7.93万
• 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 1991-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3457873
• 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、热力学和生物化学 与有效抗肿瘤相互作用相关的特性 抗生素 m-AMSA 和放线菌素 D 与核酸。 具体来说， 这些研究工作将旨在探索结构和 药物后天然和合成 DNA 的功能特性 利用配体的平衡和共价模式进行结合 相互作用并将药物结构与药物结构的关系相关联 与复杂形成相关的生物物理参数。 实验将被设计为将 m-AMSA 的结合等温线扩展到 结合药物浓度非常低，提供更准确的 其化疗药物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 通过光谱、平衡透析和相分配获得 技术。 相分配方法特别适用于 在极低的结合浓度下获得准确的结合等温线 药品。 光亲和标记技术将扩展到放线菌素 D系统。 共价连接的能力将极大地帮助 靶点特异性、协同性和/或药物聚类的研究 现象。 通过对母体放线菌素 D 进行化学修饰 光反应取代基（叠氮基部分），其功能类似物 将产生具有平衡结合特性的母体分子 与母体化合物相同（在没有光的情况下） 具有光解活化后共价连接的能力。 与放线菌素 D 的共价性质相关的特性 - 核酸加合物将通过多种生化检查进行详细检查 和生物物理技术。 将使用限制性核酸内切酶活性 监测结合位点特异性和核酸结构 加合物形成引起的扰动。 同样，几何和 放线菌素 D-DNA 加合物的分子取向将被确定 通过高场核磁共振在寡核苷酸水平 光谱学。 这些关于 m-AMSA 和放线菌素 D 的研究将增加 了解药物结合特性、药物本质 - DNA 复合物和/或加合物，并提供强大的光亲和探针 核酸的结构和功能特性的进一步定义 酸。