拉帕替尼和阿司匹林双联化合物的设计、合成及其体外活性研究  

Design,synthesize dicompound of Lapatinib and Aspirin and its in vitro activity

作　　者：袁盼盼 郭凯丽 姚琳 过晓芳[4] 刘继平 王斌 王国全 朱星枚[1] YUAN Panpan;GUO Kaili;YAO Lin;GUO Xiaofang;LIU Jiping;WANG Bin;WANG Guoquan;ZHU Xingmei(Shaanxi Key Laboratory of Traditional Medicine Foundation and New Drug Research,Shaanxi University of Chinese Medicine,Shaanxi Province,Xianyang 712046,China;Key Laboratory of Pharmacodynamics and Material Basis of Chinese Medicine of Shaanxi Administration of Traditional Chinese Medicine,Shaanxi Province,Xianyang 712046,China;Department of Medicinal Chemistry and Pharmaceutical Analysis,School of Pharmacy,Air Force Medical University,Shaanxi Province,Xi’an 710032,China;School of Science,Xi’an Technological University,Shaanxi Province,Xi’an 710021,China)

机构地区：[1]陕西中医药大学陕西省中医药基础与新药研究重点实验室,陕西咸阳712046 [2]陕西省中医药管理局中药药效与物质基础重点实验室,陕西咸阳712046 [3]空军军医大学药学系药物化学与药物分析教研室,陕西西安710032 [4]西安工业大学理学院,陕西西安710021

出　　处：《中国医药导报》2023年第16期16-20,共5页China Medical Herald

基　　金：国家自然科学基金资助项目(81402186、62102304);陕西省教育厅重点实验室项目(21JS015);陕西中医药大学学科创新团队项目(2019-YL13)。

摘　　要：目的设计、合成拉帕替尼和阿司匹林双联化合物,探究其体外活性。方法采用拼合原理,以拉帕替尼和阿司匹林为原料合成孪药(CompdⅠ)。将人表皮生长因子受体2(HER2)阳性乳腺癌细胞BT474分为空白组、阿司匹林组、拉帕替尼组、CompdⅠ组。CCK-8法和平板克隆法测定半数抑制浓度(IC_(50))和细胞克隆形成率;分子对接考察CompdⅠ与AMPK/mTOR通路蛋白及环氧合酶1蛋白的结合情况;Western blot揭示AMPK/mTOR通路蛋白的表达水平。结果以拉帕替尼和阿司匹林为原料经过缩合反应合成CompdⅠ,产率为78%,熔点为101.7~102.7℃。培养24、48、72 h,CompdⅠ与拉帕替尼和阿司匹林的IC_(50)比较,差异有统计学意义(P<0.05或P<0.01)。与空白组比较,阿司匹林组、拉帕替尼组、CompdⅠ组的克隆形成率更低;与阿司匹林组、拉帕替尼组比较,CompdⅠ组的克隆形成率更低,差异有统计学意义(P<0.05或P<0.01)。CompdⅠ与表皮生长因子受体(EGFR)、HER2、AMPK、Akt1和mTOR结合能绝对值均≥5 kcal/mol,与EFGR、Akt1、mTOR对接能较拉帕替尼更低,但与环氧合酶1不能结合。与空白组比较,CompdⅠ组和拉帕替尼组EGFR、HER2、p-Akt1-Ser124、p-Akt1-Ser169、Akt1、p-mTOR及mTOR下调,阿司匹林组p-Akt1-Ser169、p-mTOR下调,CompdⅠ组p-AMPK和AMPK上调,拉帕替尼组AMPK上调,差异有统计学意义(P<0.05或P<0.01)。与拉帕替尼组比较,CompdⅠ组p-AMPK上调,差异有高度统计学意义(P<0.01)。结论CompdⅠ有更强的抗HER2阳性乳腺癌增殖活性,其主要通过激动AMPK,抑制HER2/mTOR通路发挥作用。Objective To design,synthesize dicompound of Lapatinib and Aspirin,and investigate its activity in vitro.Methods Twin drug(CompdⅠ)was synthesized by using Lapatinib and Aspirin as raw materials.Human epidermal growth factor receptor 2(HER2)positive breast cancer cells BT474 were divided into blank group,Aspirin group,Lapatinib group,and CompdⅠgroup.CCK-8 method and plate cloning method were used to determine half inhibitory concentration(IC_(50))and cell clone formation rate;the binding of CompdⅠto AMPK/mTOR pathway protein and cyclooxygenase 1 protein was investigated by molecular docking;the expression level of AMPK/mTOR pathway protein was revealed by Western blot.Results CompdⅠwas synthesized from Lapatinib and Aspirin by condensation reaction,and the yield was 78%,melting point was 101.7-102.7℃.The IC_(50) of CompdⅠwas significantly different from that of Lapatinib and Aspirin for 24,48 h,and 72 h(P<0.05 or P<0.01).Compared with blank group,the clone formation rate in Aspirin group,Lapatinib group,and CompdⅠgroup was lower,compared with Aspirin group and Lapatinib group,the clone formation rate in CompdⅠgroup was lower,and the differences were statistically significant(P<0.05 or P<0.01).The binding energy absolute values of CompdⅠwith epidermal growth factor receptor(EGFR),HER2,AMPK,Akt1,and mTOR were all≥5 kcal/mol,the binding energy of CompdⅠwith EFGR,Akt1,and mTOR was lower than that of Lapatinib,but it could not bind with cyclooxygenase 1.Compared with blank group,EGFR,HER2,p-Akt1-Ser124,p-Akt1-Ser169,Akt1,p-mTOR,and mTOR were down-regulated in CompdⅠgroup and Lapatinib group,and p-Akt1-Ser169,p-mTOR were down-regulated in Aspirin group,p-AMPK and AMPK were up-regulated in CompdⅠgroup,and AMPK was up-regulated in Lapatinib group,and the differences were statistically significant(P<0.05 or P<0.01).Compared with Lapatinib group,P-AMPK was up-regulated in CompdⅠgroup,and the difference was highly statistically significant(P<0.01).Conclusion CompdⅠhas stronger anti-HER2 prolifer

关 键 词：阿司匹林 拉帕替尼 孪药 分子对接 HER2阳性乳腺癌 

分 类 号：R914.5[医药卫生—药物化学]