灵芝酸A-靛红拼合物的合成、抗肿瘤活性及靶点预测研究

目的 将灵芝三萜中的主要成分灵芝酸A与具有抗肿瘤作用的靛红片段进行拼合，对所得的拼合物进行体外抗肿瘤活性评价，分析构效关系，并对灵芝酸A及其拼合物的靶点进行预测。方法 将灵芝酸A与1个单元或3个单元长度的聚乙二醇连接，再通过点击化学反应与靛红片段进行拼合，采用噻唑蓝（MTT）法考察合成拼合物对乳腺癌MCF-7细胞、肝癌HepG2细胞、骨肉瘤SJSA-1细胞和正常细胞系HK-2细胞的抗增殖活性。结果 共合成16个不同长度及不同取代的灵芝酸A-靛红拼合物，确定结构，为结构全新的化合物。部分拼合物表现出良好的抗肿瘤活性，且靛红片段上的取代基对抗肿瘤效果影响较大，其中6位取代活性最优。反向找靶及分子对接结果表明，灵芝酸A及拼合物可能通过调控鼠双微体2/X蛋白（mouse double minute 2/X,MDM2/X）发挥抗肿瘤作用。结论 该系列化合物对MCF-7细胞的选择性较强，化合物A11和A16具有进一步研究的价值，具有开发为双靶点或多靶点抗肿瘤化合物的潜力。

Synthesis, antitumor activity and target prediction of ganoderic acid A-isatin conjugate

Objective To combine the main component of triterpenoid in Chinese traditional and precious nourishing medicine Ganoderma lucidum,ganoderic acid A (GAA), with anti-tumor compound isatin, evaluate the anti-tumor activity of the obtained conjugate in vitro, analyze the structure-activity relationship, and predict the targets of GAA and these conjugate. Methods GAA was connected with polyethylene glycol with one or three unit lengths, and then combined with isatin by click reaction. The anti-proliferation activity of the synthesized compounds on MCF-7, HepG2, SJSA-1 and normal cell line HK-2 was investigated by MTT assay. Results A total of 16 new GAA-isatin conjugates with different lengths and different substitutions were synthesized and their structures were determined. Some of the compounds showed good anti-tumor activity, and the substitution of the isatin fragment had a great impact on the anti-tumor effect, especially in the 6-position. Target fishing and molecular docking results showed that GAA and its conjugate may exert anti-tumor effect through regulating mouse double minute 2/X (MDM2/X).Conclusion The series of the compounds have strong selectivity for MCF-7. Compounds A11 and A16 have the value of further research and have the potential to develop a dual-target or multi-target anti-tumor compound.