摘要
目的 以CA-4为先导化合物,按照微管蛋白和PI3K双靶点活性进行设计、合成一种CA-4类衍生物[1-(3,5-二氟苯基)-2-(6-甲氧基-2-(1-(3,4,5-三甲氧基苯基)乙烯基)苯并呋喃-5-基)乙烯]。并在人三阴性乳腺癌细胞MDA-MB-231细胞上进行PI3K活性初步研究。方法 采用MTT法测定IC50值,倒置显微镜观察细胞状态,流式细胞术检测细胞周期阻滞情况,AutodockTools系统进行衍生物和PI3K分子对接,ELISA法检测细胞上清液中PI3K分泌表达量,RT-PCR和Western blot分别检测PI3K mRNA和蛋白表达水平。结果 该衍生物对MDA-MB-231细胞的IC_(50)=5.78 μmol·L^(-1),使细胞内出现一系列凋亡形态变化,阻滞细胞生长周期于G_(2)期,与PI3K良好结合,结合自由能为-36.4008 kJ·mol^(-1),降低PI3K分泌量且下调PI3K的基因和蛋白表达水平。结论 该衍生物的PI3K靶点活性得到了初步验证,也可为此类化合物进一步机制研究提供参考。
Objective To design and synthesize a CA-4 derivative[1-(3,5-difluorophenyl)-2-(6-methoxy-2-(1-(3,4,5-trimethoxyphenyl)vinyl)benzofuran-5-yl)ethylene]based on the microtubule and PI3K protein as dual targets,using CA-4 as the lead compound,and to preliminarily study the PI3K activity was on human triple negative breast cancer cell line MDA-MB-231.Methods MTT was used to measure IC_(50) value,inverted microscope to observe the cell state,flow cytometry to detect cell cycle arrest,AutodockTools system to dock the derivative and PI3K molecule,ELISA to detect PI3K secretion in the cell supernatant,and RT-PCR and Western blot to detect PI3K mRNA and-protein expression levels,respectively.Results The IC50 was 5.78μmol·L^(-1).Many morphological apoptotic changes occurred,and the cell growth cycle was hindered in G_(2) phase.Derivatives bound-well with PI3K(-36.4008 kJ·mol^(-1)),the PI3K secretion decreased,and gene and protein expression levels were down-regulated.Conclusion The PI3K target activity of the derivative has been verified,providing a reference for further mechanism research of this type of derivative.
作者
高祎婷
陈嘉媛
王晓花
周昊天
GAO Yi-ting;CHEN Jia-yuan;WANG Xiao-hua;ZHOU Hao-tian(School of Pharmacy,Xinjiang Second Medical College,Karamay Xinjiang 834000)
出处
《中南药学》
CAS
2024年第5期1135-1140,共6页
Central South Pharmacy
基金
新疆维吾尔自治区自然科学基金地州科学基金项目(No.2021D01F40)
新疆第二医学院科研基金面上项目(No.MS202301)。
关键词
CA-4
三阴性乳腺癌
PI3K
combretastatin A-4
triple negative breast cancer
PI3K
