白头翁总皂苷及其主要成分抗肿瘤药效及其构效关系的研究

Study on Antitumor Effect and Its Structure-Activity Relationship of Pulsatilla Saponin and Its Major Constituents

目的:研究探讨白头翁总皂苷及其4种主要成分的抗肿瘤活性,分析其构效关系。方法:采用体外对人肝癌细胞SMMC-7721和人结肠癌细胞HT-29进行体外抑瘤实验;采用人肝癌H22移植荷瘤小鼠进行体内抑瘤实验,观察分析白头翁总皂苷及其4种主要成分的抗癌药效。结果:体外抑瘤实验中,各主要成分均显示出明显的抑瘤活性,其中以白头翁皂苷B7抑瘤活性最强,其IC50为2.64μg·mL^(-1)(SMMC-7721细胞)和2.57μg·mL^(-1)(HT-29细胞),其他依次为白头翁皂苷B3、B9和B11。白头翁总皂苷也表现出了一定的抑瘤活性。体内抑瘤实验结果与体外实验结果基本一致,只是白头翁总皂苷在整体动物实验药效更强,提示白头翁总皂苷在体内复杂环境中的抑瘤作用更明显,体现了多成分多靶点作用的特点。4个单体构效分析表明,白头翁苷元上的23位羟基化不能增强抑瘤活性,24位侧链直链式糖活性低于非直链连接的糖,直链连接中4糖数量(葡萄糖-葡萄糖-鼠李糖-阿拉伯糖)(白头翁皂苷B9)的活性强于2糖的数量(鼠李糖-阿拉伯糖)(白头翁皂苷B11)。结论:白头翁皂苷成分中23位不是活性位点(药效团),24位所连接的糖的数量及其连接方式对其抑瘤活性有一定的影响。白头翁总皂苷体内活性强于体外的实验,提示总皂苷中其他成分在体内复杂环境下具有抑瘤的协同效应。

Objective： To study the anti-tumor activity of Pulsatilla saponins and its four main components,and analyze its structure-activity relationship. Methods： Human hepatocellular carcinoma cell line SMMC-7721 and human colon carcinoma cell line HT-29 were treated with anti-tumor activity in vitro. The tumor-bearing mice were transplanted with H22 hepatocellular carcinoma cell line to observe the effect of Pulsatilla saponin and its major components on anti-cancer effect. Results：In vitro anti-tumor experiment,all the major components showed obvious anti-tumor activity,of which anemoside B7 had the strongest anti-tumor activity with IC50 of 2. 64 μg·mL^-1（SMMC-7721 cells） and 2. 57 μg·mL^-1（HT-29 cells）,others were anemoside B3,B9 and B11. Pulsatilla saponin also showed some anti-tumor activity. Both in vivo and in vitro experimental results are basically the same. Only Pulsatilla total saponins in the mouse experiments displayed more effective than others,implying Pulsatilla total saponins in complex environments in vivo showed anti-tumor effect distinctly,which reflected the characteristics of multi-component and multi-target role. The structure-activity analysis of four components showed that the hydroxylation at C-23 of aglycon did not enhance the antitumor activity. The linearity of the straight monosaccharide chain attached to C-24 of the aglycon was lower than that of the non-linear-monosaccharide chain. The number of 4 monosaccharides（ glucose-dextroserhamnose-arabinose）（ anemoside B9） in the straight-chain linkage was more active than the amount of 2-glycolic chain（ rhamnose-arabinose）（ anemoside B11）. Conclusion： C-23 of aglycon of anemosides is not active essential for anti-cancer（ pharmacophore） effect. The amount of monosaccharide chain and the manner of its attachment to C-24 have a certain impact on its antitumor activity. Pulsatilla saponin in vivo activity is stronger than in vitro experiments suggests that the other components of the total saponins in vivo complex environment with anti-tumor synergistic effect.