TUBULIN ASSOCIATIONS AND EFFECTS OF ANTICANCER DRUGS

拟议研究的目的是为了详细了解 在分子（和亚分子）水平上， 抗有丝分裂药物与微管蛋白结合并诱导其聚合 在每种情况下对特定结构的反应。 待研究药物 是秋水仙碱、长春碱和紫杉醇，以及它们的类似物， 衍生物和对应于复合物部分的较小分子 药物结构。 具体而言，药物结合和 微管蛋白的自缔合或构象变化将被表征 通过严格的热力学和动力学方法。 之间的联系 同一药物分子的不同部分与微管蛋白的相互作用 使用小分子进行检查，如文多灵和cathetrin 长春碱的例子 的约束力的相互影响， 单独的药物相互作用之后将进行热力学和 构象技术来表征协同性（正，如 在秋水仙碱-长春碱中，或在秋水仙碱-紫杉醇中为阴性），其 存在于它们的绑定中。 药物结合位点的相对位置 对微管蛋白分子和它们的位移诱导的结合其他 药物将通过测量药物和药物之间的距离来确定。 微管蛋白上的参考点，如可交换位点GTP，以及 药物分子和这些分子的一部分之间的联系。 这些距离将 可通过NMR、EPR和荧光能量转移方法测量。 为此 目的、适当的氟化、自旋标记和荧光标记 基团将被引入药物的结构中， 类似物 所使用的方法将是那些高分子物理 生物化学，包括沉降，荧光分光光度法，密度测定法， 定量凝胶色谱，光散射，电泳，傅立叶 变换红外光谱、圆二色谱、核磁共振和电子顺磁共振，以及 如化学修饰和合成。 这项研究旨在 了解某些抗癌药物的作用方式 他们的功能，希望确定结构特征， 在设计新的此类药物中是有用的。 （左）

The aims of the proposed research are to arrive at a detailed understanding of the mechanism on the molecular (and submolecular) level of how certain anti-mitotic drugs bind to tubulin and induce it to undergo polymerization reactions to particular structures in each case. The drugs to be studied are colchicine, vinblastine and taxol, as well as their analogs and derivatives, and smaller molecules which correspond to parts of the complex drug structures. Specifically, the linkages between drug binding and self-association or conformational changes of tubulin will be characterized by rigorous thermodynamic and kinetic approaches. The linkage between the interactions of different parts of the same drug molecule with tubulin will be examined by using small molecules, such as vindoline and catharantine in the case of vinblastine. The mutual effects of the binding of the individual drugs on each other will be followed by thermodynamic and conformational techniques to characterize the cooperativity (positive, as in colchicine-vinblastine, or negative as in colchincine-taxol) which exists in their bindings. The relative locations of the drug binding sites on the tubulin molecule and their shifts induced by the binding of other drugs will be determined by measuring distances between the drugs and a reference point on tubulin, such as the exchangeable site GTP, as well as between drug molecules and parts of these molecules. These distances will be measured by nmr, epr and fluorescence energy transfer methods. For this purpose, appropriate fluorinated, spin-labelled and fluorescent marker groups will be introduced into the structures of the drugs and their analogs. The methods used will be those of macromolecular physical biochemistry, including sedimentation, spectrofluorimetry, densimetry, quantitative gel chromatography, light scattering, electrophoresis, Fourier transform infra-red spectroscopy, circular dichroism, nmr and epr, as well as chemical modification and synthesis. This research is aimed at an understanding of the manner in which certain anti-cancer drugs perform their function with the hope of identifying structural features which will be useful in the design of new such drugs. (L)