齐墩果酸氮杂环衍生物合成及对非小细胞肺癌A549作用研究

Synthesis of Oleanolic Acid Nitrogen Heterocyclic Derivative and Evaluation of its Anti-NSCLC Activity in Vitro

目的:开展齐墩果酸氮杂环衍生物合成及抗非小细胞肺癌A549作用研究。方法:以齐墩果酸为原料,通过氧化、鲍奇还原胺化、缩合等反应合成齐墩果酸氮杂环衍生物。采用CCK-8试剂盒、流式Annexin V-FITC/PI双染色法和Caspase-3试剂盒分别检测A549细胞的增殖抑制作用、细胞凋亡及Caspase-3的变化。结果:成功合成齐墩果酸氮杂环衍生物3-N-(3,5,6-三甲基吡嗪-2-甲酰基)-齐墩果酸,产率为71.2%。齐墩果酸氮杂环衍生物对A549细胞具有抑制作用且呈量效关系,IC50为110μmol·L^(-1),CC50为146μmol·L^(-1)。齐墩果酸氮杂环衍生物能诱导A549细胞株的凋亡,显著增强A549细胞中Caspase-3的活性。结论:合成的齐墩果酸氮杂环衍生物结构为3-N-(3,5,6-三甲基吡嗪-2-甲酰基)-齐墩果酸,该化合物对A549细胞株具有增殖抑制作用。诱导A549细胞株凋亡与增强Caspase-3的酶活性可能是其机制。

Objective:To study on the synthesis of oleanolic acid nitrogen heterocyclic derivative and its anti-A549 effects in vitro.Methods:The oleanolic acid nitrogen heterocyclic derivative was synthesized from oleanolic acid(OA)by oxidation,Borch reduction amination,and condensation.Anti-proliferation and apoptosis-induced effects and the change of Caspase-3 activity of the oleanolic acid nitrogen heterocyclic derivative on A549 were respectively detected by CCK-8、flow cytometry with Annexin V-FITC/PI double staining apoptosis detection kit and Caspase-3 detection kit.Results:The new oleanolic acid nitrogen heterocyclic derivative was successfully synthesized.The yield of the reaction was 71.2%.The inhibition of oleanolic acid nitrogen heterocyclic derivatives on A549 cells was in a dose-dependent manner.IC50=110μmol·L-1,CC50=146μmol·L-1.The new oleanolic acid nitrogen heterocyclic derivative can induce apoptosis of A549 and enhance caspase-3’s activity significantly.Conclusion:The new oleanolic acid nitrogen heterocyclic derivative’s structure was confirmed as 3-N-(3,5,6-trimethylpyrazine-2-formyl)-oleanolic acid which had inhibitory effect on the proliferation of A549 cells.Meanwhile,induction of apoptosis of A549,and enhancement of the activity of Caspase-3 may be its mechanism.