新型CDK9小分子抑制剂的设计和发现  

Design and discovery of novel small-molecule inhibitor of CDK9

作　　者：秦立[1] 纪庆[1] 高瀛岱[1] 刘娟妮[1] 杨铭[1] 邵晓枫[1] 熊冬生[1] 

机构地区：[1]中国医学科学院北京协和医学院血液学研究所,天津300020

出　　处：《中国药理学通报》2009年第10期1299-1303,共5页Chinese Pharmacological Bulletin

基　　金：天津市科技计划资助项目(No08ZCKFSH04100)

摘　　要：目的通过同源模建、计算机虚拟筛选和生物活性筛选,寻找新型的CDK9小分子抑制剂。方法采用同源模建方法得到CDK9的三维晶体构象,用DOCK(分子对接)对小分子三维数据库进行虚拟筛选。采用MTT法对挑选出的化合物进行生物活性测定,然后挑选高活性的化合物进行CDK9与小分子之间相互作用的研究,验证化合物对CDK9激酶活性的抑制作用。结果用DOCK程序对三维化合物库虚拟筛选后挑选出得分高的前1000个化合物,按结构差异化分类,最终选择并购买了27个代表性化合物进行进一步生物学活性测定。27个化合物中12个化合物明显抑制肿瘤细胞的增殖,其中1个化合物C-21的半数抑制浓度(IC50)在20μmol.L-1以下。选择C-21类化合物进一步进行研究,结果显示此化合物能够有效抑制各类肿瘤细胞系的增殖。酶学水平研究表明此类化合物能够有效抑制CDK9激酶活性,并在较低浓度范围内呈明显的剂量依赖关系。结论通过同源模建、计算机虚拟筛选、生物活性实验和分子水平研究,得到了一类靶向CDK9的全新的先导化合物。Aim To look for novel small-molecule inhibitors of CDK9 through structure-based virtual screening and biological activity determination. Methods Homology modeling of CDK9 was based on the 3- D structure of other cyclin-dependent kinase family members, and then virtual screening by DOCK （molecular docking） of database of small molecule was carried on. MTT method was used in inhibition of tumor cell growth in vitro, while Western blot was used for further study of molecular mechanisms. Results From the top 1000 compounds with the best DOCK energy score,27 compounds were selected for biological assay based on the diversity of chemical structure and functional group. 12 of 27 selected compounds showed significantly inhibition activity on tumor cell proliferation,and only one compound in 12 with half-maximum inhi- bition concentration （ IC50 ） values less than 20 μmol · L^-1 named C-21 was selected for further molecular mechanism study. The western blotting data showed C- 21 compound could effectively inhibit CDK9 from phosphorylating large subunit C-terminal of RNA polymerase Ⅱ in a dose-dependent manner. Conclusions Through homology modeling, virtual screening by computer, determination of biological activity and experimental studies of molecular mechanism, a new promising lead compound targeted for CDK9 was found and confirmed.

关 键 词：CDK9 同源模建 小分子抑制剂 RNA POLYMERASE Ⅱ 磷酸化 肿瘤靶点 先导物 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R345.57[医药卫生—基础医学]