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

doi:10.20047/j.issn1673-7210.2023.16.03

摘要
图/表
参考文献(25)
相关文章 (15)

全文: PDF (1209 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

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（IC50）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IC50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。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	袁盼盼1　　郭凯丽2　　姚琳3　　过晓芳4　　刘继平2　　王斌2　　王国全2　　朱星枚1

关键词 ：阿司匹林, 拉帕替尼, 孪药, 分子对接, HER2阳性乳腺癌

Abstract：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 CompdⅠ group. CCK-8 method and plate cloning method were used to determine half inhibitory concentration (IC50) and cell clone formation rate; the binding of CompdⅠ to 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 CompdⅠ was synthesized from Lapatinib and Aspirin by condensation reaction, and the yield was 78%, melting point was 101.7-102.7℃. The IC50 of CompdⅠ was 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 CompdⅠ group was lower, compared with Aspirin group and Lapatinib group, the clone formation rate in CompdⅠ group was lower, and the differences were statistically significant (P＜0.05 or P＜0.01). The binding energy absolute values of CompdⅠ with epidermal growth factor receptor (EGFR), HER2, AMPK, Akt1, and mTOR were all ≥5 kcal/mol, the binding energy of CompdⅠ with 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 CompdⅠ group and Lapatinib group, and p-Akt1-Ser169, p-mTOR were down-regulated in Aspirin group, p-AMPK and AMPK were up-regulated in CompdⅠ group, 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 CompdⅠ group, and the difference was highly statistically significant (P＜0.01). Conclusion CompdⅠ has stronger anti-HER2 proliferative activity, which acts mainly by activating AMPK, and inhibiting HER2/mTOR pathway.

Key words： Aspirin Lapatinib Twin drug Molecular docking HER2-positive breast cancer

基金资助: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）。

通讯作者: 朱星枚（1981.6-），女，博士，副教授，硕士生导师；研究方向：抗肿瘤药物物质基础与机制。王斌（1978.9-），男，博士，教授，硕士生导师；研究方向：药理学。

作者简介: 袁盼盼（1997.5-），女，陕西中医药大学药学院2020级药学专业在读硕士研究生；研究方向：抗肿瘤药物物质基础与机制。

引用本文:

袁盼盼1　　郭凯丽2　　姚琳3　　过晓芳4　　刘继平2　　王斌2　　王国全2　　朱星枚1. 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[J]. 中国医药导报, 2023, 20(16): 16-20.
YUAN Panpan1 GUO Kaili2 YAO Lin3 GUO Xiaofang4 LIU Jiping2 WANG Bin2 WANG Guoquan2 ZHU Xingmei1. Design, synthesize dicompound of Lapatinib and Aspirin and its in vitro activity. 中国医药导报, 2023, 20(16): 16-20.

链接本文:

https://www.yiyaodaobao.com.cn/CN/10.20047/j.issn1673-7210.2023.16.03 或 https://www.yiyaodaobao.com.cn/CN/Y2023/V20/I16/16

[1] Waks AG，Winer EP. Breast Cancer Treatment：A Review [J]. JAMA，2019，321（3）：288-300.
[2] Haji F，Hurvitz SA. Can Women With HER2-Positive Metastatic Breast Cancer Be Cured? [J]. Clin Breast Cancer，2021，21（6）： 526-531.
[3] Haddad TC，He J，O Sullivan CC，et al. Randomized Phase Ⅱ Trial of Capecitabine and Lapatinib with or without IMC-A12（Cituxumumab）in Patients with HER2-Positive Advanced Breast Cancer Previously Treated with Trastuzumab and Chemotherapy：NCCTG N0733（Alliance）[J]. Breast Cancer Res Treat，2021，188（2）：477-487.
[4] Voigtlaender M，Schneider-Merck T，Trepel M. Lapatinib [J]. Recent Results Cancer Res，2018，211（8）：19-44.
[5] Wahdan-Alaswad R，Liu B，Thor AD. Targeted lapatinib anti-HER2/ErbB2 therapy resistance in breast cancer：opportunities to overcome a difficult problem [J]. Cancer Drug Resist，2020，2（3）：179-198.
[6] Loibl S，Majewski I，Guarneri V，et al. PIK3CA mutations are associated with reduced pathological complete response rates in primary HER2-positive breast cancer：pooled analysis of 967 patients from five prospective trials investigating lapatinib and trastuzumab [J]. Ann Oncol，2016，27（8）：1519- 1525.
[7] Ricciotti E，Fitzgerald GA. Aspirin in the Prevention of Cardiovascular Disease and Cancer [J]. Annu Rev Med，2021，72（1）：473-495.
[8] Joshi SN，Murphy EA，Olaniyi P，et al. The multiple effects of aspirin in prostate cancer patients [J]. Cancer Treat Res Commun，2021，26：100267-100273.
[9] Grancher A，Michel P，Di Fiore F，et al. Aspirin and colorectal cancer [J]. Bull Cancer，2018，105（2）：171-180.
[10] 朱星枚，杨嘉妮，张恩户，等.阿司匹林抗人乳腺癌增殖的直接作用蛋白靶点的生物信息析[J].南方医科大学学报，2019，39（10）：1141-1148.
[11] Mccarthy AM，Kumar NP，He W，et al. Different associations of tumor PIK3CA mutations and clinical outcomes according to aspirin use among women with metastatic hormone receptor positive breast cancer [J]. BMC Cancer，2020， 20（1）：347-356.
[12] Zhou Y，Simmons J，Jordan CD，et al. Aspirin Treatment Effect and Association with PIK3CA Mutation in Breast Cancer：A Biomarker Analysis [J]. Clin Breast Cancer，2019， 19（5）：354-362.
[13] 朱星枚.细胞自噬促进拉帕替尼抗HER2阳性乳腺癌的机制研究[D].西安：第四军医大学，2013.
[14] Om Silakari，Pankaj Kumar Singh. Concepts and Experimental Protocols of Modelling and Informatics in Drug Design [M]. Academic Press，2021：131-155.
[15] Lin R，Bao X，Wang H，et al. TRPM2 promotes pancreatic cancer by PKC/MAPK pathway [J]. Cell Death Dis，2021， 12（6）：585-596.
[16] Kronstein-Wiedemann R，Tonn T. Colony Formation：An Assay of Hematopoietic Progenitor Cells [J]. Methods Mol Biol， 2019，2017：29-40.
[17] Moradipoodeh B，Jamalan M，Zeinali M，et al. In vitro and in silico anticancer activity of amygdalin on the SK-BR-3 human breast cancer cell line [J]. Mol Biol Rep，2019，46（6）： 6361-6370.
[18] Zulkipli NN，Zakaria R，Long I，et al. In Silico Analyses and Cytotoxicity Study of Asiaticoside and Asiatic Acid from Malaysian Plant as Potential mTOR Inhibitors [J]. Molecules，2020，25（17）：3991-4002.
[19] Peiter GC，de Souza CBT，de Oliveira LM，et al. COVID-19 liver and gastroenterology findings：An in silico analysis of SARS-CoV-2 interactions with liver molecules [J]. World J Hepatol，2022，14（6）：1131-1141.
[20] Ponnusamy L，Kothandan G，Manoharan R，et al. Berberine and Emodin abrogates breast cancer growth and facilitates apoptosis through inactivation of SIK3-induced mTOR and Akt signaling pathway [J]. Biochim Biophys Acta Mol Basis Dis，2020，1866（11）：165897-165907.
[21] Zhao M，Scott S，Evans KW，et al. Combining Neratinib with CDK4/6，mTOR，and MEK Inhibitors in Models of HER2-positive Cancer [J].Clin Cancer Res，2021，27（6）：1681-1694.
[22] 贺佳楠，黄旭虎，张涵，等.抗肿瘤先导化合物990209孪药的设计合成及活性[J].中国药物化学杂志，2018， 28（5）：363-370.
[23] Zhou X，Li W，Zhou R，et al. Bioinspired Scalable Total Synthesis of Opioids [J]. CCS Chem，2021，3（9）：8.
[24] Savukaityt■ A，Gudoityt■ G，Bartnykait■ A，et al. siRNA Knockdown of REDD1 Facilitates Aspirin Mediated Dephosphorylation of mTORC1 Target 4E-BP1 in MDA-MB- 468 Human Breast Cancer Cell Line [J]. Cancer Manag Res， 2021，13（2）：1123-1133.
[25] Forgerini M，Lucchetta RC，Urbano G，et al. Genetic polymorphisms associated with upper gastrointestinal bleeding：a systematic review [J]. Pharmacogenomics J，2021，21（1）：20-36.