阿霉素脂质体制备工艺优化及节拍式给药细胞探讨

Study on Preparation of Doxorubicin Liposome and Its Cytotoxicity Evaluation with Rhythmical Low-Dosage Administration

目的:优化阿霉素脂质体处方和制备工艺,利用节拍式低剂量给药化疗方式探讨阿霉素脂质体对HO-8910人卵巢癌细胞的体外杀伤作用.方法:采用硫酸铵梯度法制备脂质体,正交设计优选制备处方;葡聚糖凝胶柱层析法分离含药脂质体与游离药物并测定包封率;考察其体外释放度.以常规给药24h和节拍式低剂量给药144h的形式分别考察阿霉素脂质体、阿霉素体外细胞的毒性作用.结果:所得优化处方为:磷脂与胆固醇的质量比为3∶1,硫酸铵浓度为155mmol/L,药物与磷脂的质量比为1∶10,孵育温度为60℃.以优化处方制得的脂质体平均包封率达92.86%;脂质体具有一定的缓释作用.脂质体的细胞毒性实验结果显示,游离阿霉素与阿霉素脂质体IC50值24h分别为0.62μg/mL和0.46μg/mL,144h分别为0.06μg/mL和0.01μg/mL.结论:优化得到的阿霉素脂质体处方工艺简便,包封率高;无论是游离的阿霉素或是阿霉素脂质体,运用节拍式低剂量给药的方式在H0-8910细胞中抑制率都明显优于MTD化疗方式给药,其中脂质体药物节拍式低剂量给药方式对H0-8910细胞的抑制率明显优于游离药物.

Objective ：To optimize the formula and preparation technique of doxorubicin liposome, and to evaluate its cytotoxicity on human ovarian cancer HO8910 cells in vitro with rhythmical low-dosage administration. Methods：Doxorubicin liposome was prepared by the （NH4 ）2SO4-gradient method. The optimum formation was selected by means of the orthogonal design of experiment .The in vitro drug release behavior of liposome was evaluated by the dialysis method. MTT assay was applied to investigate the cytotoxicity of doxorubicin liposome and free doxorubicin with common administration after 24h and rhythmical low-dosage administration after 144h on HO8910 cells .Results ：The optimum formula was as follows ：the ratio of lecithin to cholesterol was 3∶1, （NH4）2SO4 concentration was 155 mmol/L ,drug∶lecithin was 1∶10 ,temperature was 60 ℃ .The average entrapment efficiency for doxorubicin was 92.86% .The release of liosome was relatively slow. The cytotoxicity test showed that IC50 of free doxorubicin and doxorubicin liposomes were 0.62 μg/mL and 0.46 μg/mL respectively at 24h and 0.06 μg/mL and 0.01 μg/mL at 144h .Conclusion ：The selected formulation and preparation technique of doxorubicin liposome is reasonable in prescription ,practicable in techniques and high in encapsulation efficiency. MTT indicated that the inhibition rates of free doxorubicin and doxorubicin liposomes with rhythmical low-dosage administration were much higher than those with MTD chemotherapy administration, and rhythmical low-dosage administration had much higher inhibition rate for HO 8910 cells than free drug .