博落回中MELK小分子抑制剂的筛选及体外活性研究  

Virtual Screening and in Vitro Activity Study on Small-molecule MELK Inhibitors from Macleaya Cordata

作　　者：陈佩文 陈嘉烨 陈志红 龚先玲 CHEN Peiwen;CHEN Jiaye;CHEN Zhihong;GONG Xianling(School of Pharmacy,Guangdong Medical University,Dongguan 523808,China)

机构地区：[1]广东医科大学药学院,广东东莞523808

出　　处：《中国现代应用药学》2025年第4期578-588,共11页Chinese Journal of Modern Applied Pharmacy

基　　金：广东省中医药局项目(20201182、20211222);广东医科大学博士启动项目(B2019029);广东医科大学强校基金项目(4SG23006G)。

摘　　要：目的以MELK为靶标,通过虚拟筛选和体外活性实验,从中药博落回中筛选抑制结直肠癌的小分子抑制剂。方法通过文献检索博落回中的苯并菲啶类生物碱成分,确定为小分子化合物(配体),运用AutoDock Vina软件将这些化合物与MELK对接,根据打分值虚拟筛选出博落回中潜在抑制MELK激酶的化学成分。采用MTT法研究化合物对结直肠癌细胞的增殖抑制作用,Western blotting检测细胞PARP、Caspase-3和MELK蛋白的表达。细胞热转移实验检测化合物与MELK蛋白的结合能力。结果分子对接显示44个化合物能与MELK蛋白形成稳定对接,22个化合物具有较强的结合能力。体外实验结果显示6-乙氧基血根碱、二氢血根碱、6-丙酮基二氢血根碱3个化合物均能呈浓度和时间依赖性地抑制结直肠癌细胞的增殖,与MELK蛋白结合并下调其蛋白的表达;且6-乙氧基血根碱下调结直肠癌细胞PARP、Caspase-3蛋白表达,上调Cleaved PARP和Cleaved caspase-3蛋白表达。结论综合虚拟筛选和体外实验,筛选出3个抑制MELK蛋白的苯并菲啶类化合物,为研发新型的MELK小分子抑制剂治疗结直肠癌提供实验依据。OBJECTIVE To screen out small molecule MELK inhibitors on colorectal cancer through virtual screening and in vitro experiment from Macleaya cordata,a traditional Chinese medicine.METHODS The benzophenanthridine alkaloids from Macleaya cordata were collected through literature search and identified as a small molecule compound(ligand).By AutoDock Vina software,the potential compounds that inhibited MELK kinase were screened according to the score.The inhibition effect of the compounds on colorectal cancer cells was detected by using MTT assay.The expression of PARP,Caspase-3 and MELK proteins was analyzed by Western blotting.Cell thermal shift assay was used to evaluate the binding capacity of compounds with MELK protein.RESULTS Forty four compounds could stably dock with MELK protein,and 22 compounds had a relatively strong ability to bind with MELK protein.The result of in vitro experiment showed that,three compounds including 6-ethoxysanguinarine,dihydrosanguinarine and 6-acetonyldihydrosanguinarine inhibited the proliferation of colorectal cancer cells in a concentration-and time-dependent manner,bound with and down-regulated MELK protein respectively.The expression of PARP and Caspase-3 proteins was down-regulated,and the levels of Cleaved PARP and Cleaved caspase-3 were up-regulated in colorectal cancer cells treated with 6-ethoxysanguinarine.CONCLUSION By virtual screening and in vitro experiments,three benzophenanthridine compounds that inhibited MELK protein are screened,which will provide an experimental basis for the development of new small molecule MELK inhibitors for the treatment of colorectal cancer.

关 键 词：分子对接 博落回 结直肠癌 MELK抑制剂 

分 类 号：R285.5[医药卫生—中药学]