新颖asperphenamate衍生物的设计合成及组织蛋白酶抑制活性和自噬诱导作用研究  

Design and synthesis of novel asperphenamate derivatives and evaluation of autophagy induction and cathepsin inhibitory effects

作　　者：邹春阳 门金玉[3] 王垚 王凤秋[1,2] 阮馨慧 ZOU Chunyang;MEN Jinyu;WANG Yao;WANG Fengqiu;RUAN Xinhui(Department of Pharmacy,Liaoning Vocational College of Medicine,Shenyang 110101,China;Liaoning Institute of Basic Medicine,Shenyang 110101,China;Shenyang Pharmaceutical University,Shenyang 110016,China)

机构地区：[1]辽宁省基础医学研究所,辽宁沈阳110101 [2]辽宁医药职业学院药学系,辽宁沈阳110101 [3]沈阳药科大学,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2021年第12期1262-1271,共10页Journal of Shenyang Pharmaceutical University

基　　金：辽宁省科技厅研究项目(2019-MS-182)。

摘　　要：目的设计合成asperphenamate分子内C环被氰基取代的衍生物,并研究其对于组织蛋白酶(cathepsin)的抑制作用,同时评价其调控乳腺癌细胞自噬能力。方法以光学活性的天冬酰胺和苯丙氨酸为原料,将所得中间体缩合得到了44个光学活性的衍生物,所得产物均经^(1)H-NMR和HR-MS验证了结构。利用荧光底物方法,以asperphenamate为对照药,评价了所合成衍生物的组织蛋白酶抑制作用,选择酶抑制作用最优的分子,利用AO染色和蛋白免疫印迹方法(western blotting)评价其调控乳腺癌细胞MCF-7自噬的能力。结果和讨论组织蛋白酶活性评价结果表明,部分化合物显示了酶抑制能力,对组织蛋白酶L和S亚型显示了明显选择性。其中,2-氟取代衍生物CA4b对组织蛋白酶L显示了最优活性,是阳性对照药的2倍。通过AO染色和蛋白免疫印迹方法分析证实CA4b也显示了明显的诱导乳腺癌细胞MCF-7的自噬作用。Objective To design and synthesize the asperphenamate derivatives which substitute the C-ring with cyano group.To evaluate the cathepsin inhibitory effect and autophagy modulating ability of these derivatives.Methods Starting from optical active phenylalanine and asparagine,forty-four stereoisomers were obtained.All structures were characterized by ^(1)H-NMR and HR-MS.Using the method of fluorescent substrate,cathepsin inhibitory activity assay of all derivatives were carried out(asperphenamate as control).AO staining experiment and western blotting analysis were used to detect the occurrence of autophagy in MCF-7 cells.Results and Conclusions Cathepsin inhibitory activity results showed that part of derivatives displayed selective inhibition against cathepsin L and S.Among them,2-fluro analog,CA-4 b,displayed the greatest inhibitory activity,and its inhibitory capability against Cat L was 2.0-fold more potent than that of asperphenamate.And AO staining experiment and western blotting analysis validated that CA-4 b can induce autophagy in MCF-7 cells.

关 键 词：组织蛋白酶抑制剂 诱导自噬作用 asperphenamate 设计合成 抗乳腺癌作用 

分 类 号：R917[医药卫生—药物分析学]