APC/Asef相互作用抑制剂的设计、合成及其抗结肠癌活性研究

The truncated APC/Asef interaction, a protein-protein interaction, was identified and validated successfully inside the colon cancer cells as a novel druggable anti-cancer target. In previous research, we adopted APC/Asef complexes and developed a series of active peptides based on the complex structure. Then a molecular hit A1B3 was obtained as an effective APC/Asef inhibitor though the virtual screening based on the pharmacophore models and the biological evaluation of anti-colon cancer on the cellular and molecular level. Then compounds with high activity will be further designed and optimized with methods of the interaction between APC/Asef complex and 3D-QASR. Then the pharmacodynamic effects of compounds of anti-colon cancer in vivo and in vitro will be tested in order to obtain the desired anti-cancer compounds. All tasks will testify truncated APC/Asef interaction as a powerful target against colorectal cancer and provide a proof of principle for clinical therapy using this lead compound.

截短型 APC/Asef 蛋白蛋白相互作用在结肠癌的发生发展中起着重要作用。我们课题组在解析截短型APC/Asef蛋白共晶复合物（Cell Res， 2012）的基础上，发现一系列活性多肽，进而通过药效团模型的构建和大规模虚拟筛选获得了分子水平和细胞水平均可有效抑制APC/Asef相互作用的苗头化合物A1B3。本项目拟在此基础上，运用相互作用模式和三维定量构效关系相结合的方法，进一步设计、优化A1B3，以期得到更高活性的APC/Asef 相互作用小分子抑制剂，并通过系统的药理学实验综合评估其抗结肠癌活性。 这些工作的实施将为结肠癌的治疗提供新的先导化合物，可望为结肠癌的治疗提供全新的途径和方案。

过度激活的截短型APC（Adenomatous Polyposis Coli，腺瘤性息肉蛋白）和Asef（APC-stimulated guanine nucleotide exchange factor，APC刺激的鸟苷酸交换因子）之间的相互作用在结肠癌的发生发展过程中发挥着重要作用，可作为新型的结肠癌治疗靶标。我们在APC/Asef蛋白共晶复合物揭示其相互作用位点的基础上，通过相互作用模式和三维定量构效关系相结合的方法，设计合成了一系列APC/Asef相互作用抑制剂，最终发现了高活性的抑制APC/Asef相互作用的先导物MAI-203和MAI-400。MAI-203的Ki值为0.015 ± 0.02 µM，相比起始化合物活性提高3000多倍；MAI-400，其IC50高达0.2 µM，MAI-400通过分子内氢键的形成，增加了与APC结合时间并提高活性。MAI-203和MAI-400均可以选择性阻断APC/Asef相互作用，抑制其下游CDC42的GTPase酶活性，从而阻断结肠癌的恶性转移。这不仅为结肠癌的治疗提供新的靶点和先导化合物，也为结肠癌的治疗提供全新的途径和方案。