表没食子儿茶素没食子酸酯烷基化衍生物合成及其体外抗癌活性

目的:设计合成表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)烷基化程度较高的衍生物,对其进行简单构效分析,并对衍生物抑制肝癌细胞增殖的药理活性进行初步筛选,旨在选出结构较稳定药效较好的EGCG衍生物。方法:以EGCG,(CH_3CH_2O)_2SO_2为原料,通过乙基化反应合成EGCG衍生物,采用噻唑蓝(MTT)法,细胞密度1×10~8/m L,将EGCG衍生物3,4稀释成4个浓度,对SMMC-7721细胞其质量质量浓度分别为60,80,100,150 mg·L~(-1)和30,40,50,60 mg·L~(-1),对Hep G2细胞其质量浓度为60,80,100,150 mg·L~(-1)和20,30,40,50 mg·L~(-1),作为药物组,并设空白组,每个浓度组设4个复孔,加药后继续培养24 h,测定乙基化EGCG对肝癌细胞SMMC-7721,Hep G2的影响。结果:合成4个EGCG衍生物,其结构通过~1H-NMR,~(13)C-NMR,~2DNMR,质谱等方法进行结构鉴定,均为未见文献报道的新化合物,乙基化EGCG产物3,4对肝癌细胞SMMC-7721,Hep G2增殖均有抑制作用。结论:乙基化EGCG产物3,4对肝癌细胞SMMC-7721,Hep G2均有抑制作用,其中4的抑制作用比EGCG明显增强。

Synthesis of Epigallocatechin Gallate Alkylated Derivatives and Their Anti-tumor Activity in Vitro

Objective:To design the synthesis of high epigallocatechin gallate (EGCG) alkylated derivatives, analyze the structure-activity relationship and the pharmacological activity in inhibiting human hepatocellular carcinoma cell proliferation, in order to select the more stable and effective EGCG derivatives. Method:With EGCG and (CH₃ CH₂ O)₂ SO₂ as raw materials, a series of EGCG derivatives were ethylated. The effect of ethylated EGCG on the proliferation of human hepatocellular carcinoma cell SMMC-7721 and HepG2 were determined by 3-(4,5 dimethylthiazol-2-yl)-2,5 dimethyl-2H-tetrazolium bromide (MTT) method. With the cell density at 1×10⁸/mL, the 3/4 of EGCG derivatives were diluted into four concentrations, with 60, 80, 100, 150 mg·L^-1, and 30, 40, 50, 60 mg·L^-1 for SMMC-7721 cells, and 60, 80, 100, 150 mg·L^-1, and 20, 30, 40, 50 mg·L^-1 for HepG2 cells, both were drug groups. A cell control group was also set up. Each concentration group has four complex wells, and the cells were continuously cultured for 24 h after addition of drugs. Result:We synthesized four EGCG derivatives. Their structures were identified by ¹H-NMR,¹³C-NMR,²DNMR, mass spectrometry and other methods, all of them were novel compounds. Ethylated EGCG product 3 and 4 can inhibit proliferation of hepatocellular carcinoma cell SMMC-7721 and HepG2. Conclusion:Ethylated EGCG product 3 and 4 have inhibition effect on hepatocellular carcinoma cell SMMC-7721 and HepG2, and 4 has a much stronger effect than EGCG.