选择性识别端粒蛋白p65的苯并吡喃新先导：优化设计合成及作用机制

端粒酶在癌细胞中特异性高表达，是一种广谱的抗癌靶标。但由于端粒酶结构复杂，一直没有筛选出高效抑制剂。p65是合成端粒酶的主要蛋白。为了拓展靶标端粒酶的研究新思路，本申请基于调控端粒酶的合成过程，拟设计小分子靶向p65，从而靶标端粒酶的装配过程，阻断其合成，抑制癌细胞过度增殖。借助结构生物学对p65的最新认识，本申请在前期苯并吡喃抑制癌细胞增殖初步构效关系的基础上，依据p65蛋白晶体结构和关键残基, 利用计算机辅助设计，结合其它方法，开展基于靶标与机制的分子设计。定向合成新型苯并吡喃及五环三萜化合物库，然后进行细胞增殖及分子水平蛋白活性筛选。构建高效的三维定量构效关系，开展先导定向优化，最终筛选出高选择性靶标端粒酶装配过程的新先导。Co-IP与残基突变实验对靶标选择性及活性位点进行确认。培养先导化合物与p65复合物共晶，解析复合物精细结构；Western blotting探讨其初步作用机制。

Telomerase is a broad spectrum anticancer target which specific express in tumor cells. Because telomerase complex structure, no efficient inhibitors have not been designed. p65 is a major protein of synthesis telomerase, in order to terminate the abnormal function of telomerase activity, this study will design small molecules targeting p65, aims to target assembly process of telomerase, blocking its synthesis. Based on the latest understanding p65 function with structural biology and the basis of the already structure-activity relationships between compounds benzopyran derivatives and inhibition activity of tumor cell proliferation, we will carry out this plan. Furthermore, based on p65 protein crystal structure and key residues, using computer-aided design, in combination with other methods, rational molecular will be designed in this study. Then, a serials novel benzopyran compounds will be directed synthesized, and the compound library with structural diversity will be built. At last, cell proliferation of molecular levels and telomerase inhibition activity is to screened. Based on structural optimization OF 3D-QSAR, the initial active site recognition will been screened by Co-IP and residues mutated experiments, followed by incubation with highly active compounds eutectic p65 complexes, parsing the fine structure of the active site; Western blotting discuss its preliminary mechanism. I hope the novel lead compound with high selectivity targeted assembly process of telomerase will be found.