注射用多西他赛脂质体的制备、体外释放及细胞毒作用

Preparation,in vitro release and cytotoxic effect of docetaxel liposomes for intravenous injection

目的:制备注射用多西他赛脂质体,并考察其理化性质、体外释放特性及细胞毒作用。方法:采用薄膜分散高压均质结合冻干工艺制备多西他赛脂质体,冻干曲线为预冻温度-45℃,保温4h,第1次干燥温度-30℃,保温30h,第2次干燥25℃,保温10h;考察脂质体形态、粒径分布、Zeta电位和稳定性,过滤-超滤离心法测定脂质体的包封率;以多西他赛注射液为对照,比较了体外释放、血浆蛋白结合率及其体外细胞毒作用。结果:多西他赛脂质体的平均粒径为(85.3±4.8)nm,Zeta电位-(51.5±2.1)mV,包封率为(99.89±0.06),5葡萄糖注射液稀释后8h内稳定;在1mol/L水杨酸钠-磷酸盐缓冲液中24h累积释放约85,血浆的存在使释放所有减少;多西他赛脂质体对肿瘤细胞(HepG2、BXPC-3、SK-OV-3)的IC50均较注射液低。结论:制得的注射用多西他赛脂质体载药量和包封率均较高,粒径均匀,稳定性良好,体外细胞毒作用较注射液强。

Aim： To prepare docetaxel （DTX） liposomes for intravenous injection and investigate its release characteristics in vitro as well as its cytotoxic effect. Methods： Docetaxel liposomes were prepared by film dispersionhigh pressure homogenization. The lyophilization cycle consisted of cooling the solution down to -45 ℃ for 4 h, then drying at - 30℃for 30 h, and furthermore drying at 25℃ for 10 h; The shape of liposomes, particles size, Zeta potential and stability were evaluated. Entrapment efficiency was determined by filtration-centrifugal ultrafiltration. The in vitro release, plasma protein binding and cytotoxicity were investigated between liposomes and injections. Results： The particle size, Zeta potential and entrapment efficiency of the liposomes were （85.3 ±4. 8） nm, - （51.5 ± 2. 1） mV, （ 99.89 ± 0. 06） %, respectively. The liposomes after diluted by 5% glucose solution was stable in 8 h. The in vitro cumulative released in 1 mol/L sodium salicylate solution-PBS reached about 85 % in 24 h, and decreased in the presence of plasma. The IC50 values of DTX liposomes, evaluated on HepG2, BXPC- 3 and SK-OV-3 cell lines were lower than injection. Conclusion： Docetaxel liposomes for intravenous injection show high drugloading capability, high encapsulation efficiency and long-term stability, and liposomes can increase the cytotoxic effect compared with injection.