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目的制备硬脂醇半乳糖苷修饰的阿西替尼脂质体,并进行处方筛选及体外活性研究。方法采用硫酸铵梯度法制备硬脂醇半乳糖苷修饰的阿西替尼脂质体,以包封率及粒径为评价指标,采用Box-Behnken响应面设计法优化制备工艺,并研究硬脂醇半乳糖苷修饰的阿西替尼脂质体对人肝癌SMMC-7721细胞株的增殖抑制及凋亡的诱导作用。采用CCK-8法检测硬脂醇半乳糖苷修饰的阿西替尼脂质体对人肝癌SMMC-7721细胞株的生长抑制情况,采用了Annexin V/PI流式细胞分析法检测细胞凋亡。结果最佳工艺:药与磷脂比为1∶14.95,胆固醇与磷脂之比为1∶4.45,水浴温度为61.93℃,硫酸铵溶液的体积为4.31 m L。硬脂醇半乳糖苷修饰的阿西替尼脂质体的粒径为(252±6.4)nm,包封率为(68.50±0.85)%。CCK-8细胞毒性试验结果显示,在药物浓度相同时,抑制率随时间的延长而增加;作用时间相同时,抑制率随药物浓度的增大而增加。Annexin V/PI流式试验结果显示,硬脂醇半乳糖苷修饰的阿西替尼脂质体对人肝癌SMMC-7721细胞株的抑制率优于人肝癌A549细胞株。结论硫酸铵梯度法制备硬脂醇半乳糖苷修饰的阿西替尼脂质体的处方合理,工艺可行,包封率高。硬脂醇半乳糖苷修饰的阿西替尼脂质体对人肝癌SMMC-7721细胞株有更高的细胞毒性,诱导SMMC-7721细胞株凋亡能力比A549的强。初步判定硬脂醇半乳糖苷修饰的阿西替尼脂质体具有主动肝靶向性。
Objective To prepare stearyl alcohol galactoside modified axitinib liposomes, and the prescription screening and in vitro activity. Methods Aicitinib liposomes modified by stearyl alcohol galactoside were prepared by ammonium sulfate gradient method. The encapsulation efficiency and particle size were used as evaluation indexes. The optimal preparation technology was optimized by Box-Behnken response surface design method. Proliferation Inhibition and Apoptosis Induction of Axinib Liposome Modified with Alcohol - galactoside on Human Hepatoma SMMC - 7721 Cell Line. CCK-8 method was used to detect the growth inhibition of human hepatocellular carcinoma SMMC-7721 cells treated with stearyl galactoside modified liposomes. Cell apoptosis was detected by Annexin V / PI flow cytometry. Results The optimum process was as follows: the ratio of drug to phospholipid was 1:14.95, the ratio of cholesterol to phospholipid was 1: 4.45, the temperature of water bath was 61.93 ℃, and the volume of ammonium sulfate solution was 4.31 m L. The particle size of steatolactam modified axitinib was (252 ± 6.4) nm and the encapsulation efficiency was (68.50 ± 0.85)%. CCK-8 cytotoxicity test results showed that at the same drug concentration, the inhibition rate increased with time; the same time, the inhibition rate increases with the increase of drug concentration. Annexin V / PI flow cytometry results showed that stearyl alcohol galactoside modified axitinib liposomes on human hepatoma SMMC-7721 cell line inhibition rate is better than the human hepatocellular carcinoma A549 cell line. Conclusion The preparation of stearyl alcohol galactoside modified axitinib liposomes by ammonium sulfate gradient method is reasonable, the process is feasible and the entrapment efficiency is high. Stearyl alcohol galactoside modified axitinib liposomes on human hepatoma SMMC-7721 cell line has higher cytotoxicity, SMMC-7721 cell apoptosis induced than A549. The initial determination of stearyl galactoside modified axitinib liposomes with active liver targeting.