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目的设计合成一系列一氧化氮(NO)供体型苯胺嘧啶类化合物,以寻找活性较强的抗非小细胞肺癌(NSCLC)药物。方法将NO供体呋咱N-氧化物与苯胺嘧啶片段杂合制得目标化合物;采用MTT法考察目标物的抗肿瘤细胞增殖活性,采用Western免疫印迹法研究目标物对表皮生长因子受体(EGFR)及其下游信号通路的影响,采用Griess法测定目标化合物在肿瘤细胞中的NO释放。结果与结论目标化合物的结构经MS、~1H-NMR谱确证;生物活性测试结果表明,目标物对EGFR二次突变的H1975细胞具有不同程度的增殖抑制活性(IC_(50)=0.078~2.332μmol·L~(-1)),明显优于抗NSCLC药物吉非替尼(IC_(50)=8.589μmol·L(-1)),其中,目标物6f可选择性地抑制EGFR突变细胞的增殖,其活性(IC_(50)=0.078μmol·L(-1))与文献报道的强效化合物WZ4002(IC_(50)=0.064μmol·L(-1))相当;此外,6f还能选择性地在EGFR突变的NSCLC细胞中释放高浓度的NO,对EGFR及其下游信号AKT、ERK的磷酸化具有明显的抑制作用,故值得进一步研究。
Objective To design and synthesize a series of nitric oxide (NO) donor aniline pyrimidine compounds for the search of active anti-non-small cell lung cancer (NSCLC) drugs. Methods The target compound was obtained by heterozygous coupling of NO donor furazan N-oxide and aniline pyrimidine fragment. The anti-tumor cell proliferation activity of the target substance was investigated by MTT assay. The effect of the target substance on the expression of epidermal growth factor receptor EGFR) and its downstream signaling pathways, the NO release of the target compounds in tumor cells was determined by Griess method. RESULTS AND CONCLUSIONS The structures of the target compounds were confirmed by ~ 1H-NMR spectroscopy. The biological activity tests showed that the target compounds exhibited different degrees of proliferation inhibitory activity on H1975 cells with EGFR secondary mutation (IC 50 = 0.078-2.332 μmol · L -1), which was significantly better than that of NSFLC (IC 50 = 8.589 μmol·L -1), in which target 6f selectively inhibited the proliferation of EGFR mutant cells (IC 50 = 0.078 μmol·L -1) was comparable to that reported in the literature for the potent compound WZ4002 (IC 50 = 0.064 μmol·L -1). In addition, 6f was also able to selectively Release of high concentration of NO in EGFR mutant NSCLC cells has obvious inhibitory effect on the phosphorylation of EGFR and its downstream signals AKT and ERK and so it deserves further study.