PROBING MICROTUBULES WITH PACLITAXEL ANALOGS

描述：这是一个续订请求，旨在继续工作，以确定 微管蛋白上与抗肿瘤药物紫杉醇相互作用的位点 通过使用光亲和和亲电子类似物合成 在共同主要研究者的实验室，研究微管蛋白- 紫杉醇相互作用使用荧光类似物也制备在 共同研究者的实验室。 在不同位置具有反应性基团的类似物 紫杉醇分子周围的蛋白质， 鉴定紫杉烷分子周围结合域中的肽。 合成了2-和3 ′-N-苯甲酰基的叠氮衍生物 并发现具有良好的生物活性。 这些衍生品已经 以放射性标记的形式合成，并用于光标记 问题研究 建议合成额外的反应性衍生物 在7、10和3 '-位上，并使用它们来标记微管蛋白。 后 标记，肽是通过化学和蛋白水解过程产生的 并通过组合色谱法纯化。 序列和位点 将通过气相测序和质量测定来确定改性的量。 光谱法 作为对光诱导标记实验的补充， 紫杉醇上含有亲电取代基的几种衍生物 将用于标记微管蛋白上的结合结构域。 最后，新 制备的荧光类似物将用于评价极性 结合结构域和结合的紫杉醇暴露于 溶剂基质 对紫杉醇-微管蛋白的进一步了解 相互作用应该有助于开发新的衍生物 具有更好的活性和更低的毒性。

DESCRIPTION: This is a renewal request to continue work to identify the sites on tubulin which interact with the antitumor drug paclitaxel through the use of photo affinity and electrophilic analogs synthesized in the Co-Principal Investigator s laboratory and to study tubulin- paclitaxel interactions using fluorescent analogs also prepared in the Co-PI s laboratory. Analogs with reactive groups at different positions around the paclitaxel molecule are expected to allow the researchers to identify peptides in the binding domain around the taxane molecule. Azido derivatives of the 2-and 3'-N-benzoyl groups have been prepared and found to have good biological activity. These derivatives have been synthesized in radio-labeled form and are being used in photo labeling studies. It is proposed to synthesize additional reactive derivatives at the 7,-10-and 3'-positions and use those to label tubulin. After labeling, peptides are produced by chemical and proteolytic procedures and purified by a combination of chromatographies. Sequences and sites of modification will be determined by gas phase sequencing and mass spectrometry. As a compliment to the photo-induced labeling experiments, several derivatives containing electrophilic substituents on paclitaxel will be utilized to label the binding domain on tubulin. Finally, newly prepared fluorescent analogs will be utilized to evaluate the polarity of the binding domain and the extent bound paclitaxel is exposed to the solvent matrix. A greater understanding of paclitaxel-tubulin interactions should be helpful for the development of new derivatives with better activity and less toxicity.