一种新型PAK1抑制剂诱导结直肠癌DLD-1细胞凋亡的机制研究  

A mechanism research of novel inhibitor of PAK1 inducing colorectal cancer DLD-1 apoptosis

作　　者：王佳祺 陈姣[1,2] 孙晓艳[1,2] 卢悟广[1,2] 杨洋[1,2] 蔡雪婷[1,2] 王小宁[1,2] 曹鹏[1,2] WANG Jiaqi;CHEN Jiao;SUN Xiaoyan;LU Wuguang;YANG Yang;CAI Xueting;WANG Xiao-ning;CAO Peng(Nanjing University of Chinese Medicine,Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing 210028;Jiangsu Province Academy of Traditional Chinese Medicine,Laboratory of Cellular and Molecular Biology,Nanjing 210028,China)

机构地区：[1]南京中医药大学附属中西医结合医院,南京210028 [2]江苏省中医药研究院细胞与分子生物学实验室,南京210028

出　　处：《中国药科大学学报》2018年第2期229-237,共9页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.81573665;No.81602733)~~

摘　　要：PAK1蛋白激酶在肿瘤的发生发展中发挥着重要作用,筛选开发新的PAK1抑制剂作为肿瘤治疗靶向药物具有重要意义。传统的PAK1抑制剂筛选方法存在成本高、效率低等缺陷,而计算机虚拟筛选可降低发现活性先导化合物的成本,提高筛选效率。本实验采用计算机辅助药物虚拟筛选结合Z'lyteTM高通量激酶筛选出1种PAK1靶向候选化合物,体外酶活性检测发现化合物18(K788)具有良好的PAK1抑制活性(抑制率为42.7%)。进一步通过MTT检测发现K788具有显著的PAK1阳性肿瘤杀伤活性,抑制活性优于阳性药IPA-3(inhibitor of p21-activated kinase compound-3)。采用流式细胞分析、Western blot检测发现K788能通过抑制PAK1的表达及其活化以激活caspase凋亡通路,诱导结肠癌细胞DLD-1的凋亡。K788具有临床开发应用的潜力,可作为PAK1靶向先导分子进行深入研究。PAK1 plays an important role in the development of tumors.It is of great significance to screen and develop new PAK1 inhibitors as targeted drugs for cancer treatment.The traditional PAK1 inhibitor screening method has the problems of high cost and low efficiency.Computer virtual screening can reduce the cost of finding active lead compounds and improve the screening efficiency.In this study,a kind of PAK1 candidate compound was screened by computer assisted virtual screening combined with Z′lyte TM high flux kinase screen.In vitro enzyme activity screening showed that compound 18(K788)had good PAK1 inhibitory activity(inhibition rate was 42.7%).Furtherly by MTT detection,it was found that K788 had significant PAK1 positive tumor killing activity,which was even better than the positive drug IPA-3.Flow cytometry and Western Blot showed that K788 could activate caspase apoptosis pathway and induce apoptosis of colon cancer cell DLD-1 by inhibiting PAK1 expression and activation.K788 has great potential for clinical development and application,and can be used as a PAK1 target for further research.

关 键 词：计算机辅助药物设计 PAK1 酶活 增殖抑制 促凋亡 

分 类 号：R965[医药卫生—药理学]