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目的筛选和优化转铁蛋白、叶酸共同修饰的阿霉素脂质体的处方及制备工艺,以期得到具有良好的脑胶质瘤靶向治疗作用的给药系统。方法采用薄膜分散和硫酸铵梯度法制备阿霉素脂质体。将叶酸连接至二硬脂酸磷脂酰乙醇胺-聚乙二醇2000(DSPE-PEG2000-NH2)得到DSPE-PEG2000-Folic,考察不同磷脂种类、药脂比、水化介质和载药时间,对脂质体粒径、包封率和稳定性的影响,确定脂质体的处方工艺。以大鼠的脑毛细血管内皮细胞(bEnd3)和星形胶质细胞组成体外血脑屏障(blood-brain barrier,BBB),并结合大鼠胶质瘤C6细胞,构建体外模拟胶质瘤靶向治疗的复合BBB模型。考察阿霉素脂质体在bEnd3细胞中的摄取机制和透过BBB的转运速率及对C6细胞的毒性。结果确定了DSPC作为主要磷脂组分,并以120 mmol.L 1的硫酸铵作为水化介质,药脂比为1∶1 5,载药时间选择60 min,成功制备了高包封率和稳定性的双配体脂质体。其在bEnd3细胞中摄取远大于普通脂质体(P<0.05),摄取过程受网格蛋白和小窝内陷介导的细胞内吞作用,并受转铁蛋白和叶酸的影响;同时其在BBB模型中的药物透过速率、及其进一步透过BBB后对下层C6细胞的毒性,均显著高于其他脂质体组。结论转铁蛋白和叶酸共同修饰的阿霉素脂质体具有较好的体外脑胶质瘤靶向治疗作用。
OBJECTIVE: To screen and optimize the formulation and preparation of doxorubicin liposome co-modified with transferrin and folic acid, in order to obtain a drug delivery system with good glioma targeting therapy. Methods Adriamycin liposomes were prepared by thin film dispersion and ammonium sulfate gradient method. Folic acid was connected to DSPE-PEG2000-Folic disodium stearic phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000-NH2) to investigate the effects of different phospholipid types, lipid-lipid ratio, hydration medium and drug loading time, Plastid particle size, encapsulation efficiency and stability of the liposomes to determine the prescription process. In vitro rat brain capillary endothelial cells (bEnd3) and astrocytes constitute the in vitro blood-brain barrier (BBB), and in combination with rat glioma C6 cells, in vitro mimic glioma targeting Treatment of compound BBB model. The uptake mechanism of doxorubicin liposomes in bEnd3 cells and the rate of BBB transfection and the toxicity to C6 cells were investigated. The results showed that DSPC was the main phospholipid component, and 120 mmol.L -1 ammonium sulfate was used as the hydration medium. The drug-lipid ratio was 1:15 and the drug-loading time was 60 min. High encapsulation efficiency and stability Sexual dual ligand liposomes. Its uptake in bEnd3 cells was much larger than that in normal liposomes (P <0.05), and the uptake was mediated by endoplasmic reticulum and caveolae mediators and was affected by transferrin and folic acid. At the same time, The rate of drug permeation in the BBB model and its toxicity to the underlying C6 cells after further penetrating BBB were significantly higher than those in the other liposomes. Conclusion The doxorubicin liposome co-modified with transferrin and folic acid has a good targeting effect in glioma in vitro.