PROBING MICROTUBULES WITH PACLITAXEL ANALOGS

描述：这是一个续订请求，以继续工作以识别 微管蛋白上与抗癌药物紫杉醇相互作用的部位 通过利用光亲和亲电类似物合成 在联席首席研究员S实验室，研究微管蛋白- 紫杉醇与荧光类似物的相互作用也在 S实验室联合皮埃尔。反应性基团在不同位置的类似物 紫杉醇分子周围有望允许研究人员 鉴定紫杉烷分子周围结合域中的多肽。 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。这些衍生品已经被 以放射性标记的形式合成，并用于照片标记 学习。建议合成更多的活性衍生物。 在7、-10和3‘-位置，并用这些位置标记微管蛋白。之后 标记，多肽是通过化学和蛋白质分解过程产生的 并通过组合层析进行纯化。序列和位置 将通过气相序列和质量来确定改性的 光谱分析。作为对光诱导标记实验的赞扬， 紫杉醇上含有亲电取代基的几种衍生物 将被用来标记微管蛋白上的结合域。最后，新的 准备的荧光类似物将被用来评估极性 结合区域和范围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 相互作用应有助于开发新的衍生品 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.