共载多西紫杉醇/依克立达白蛋白纳米粒的制备与评价

Preparation and evaluation of Docetaxel and Elacridar albumin nanoparticles

目的制备共载多西紫杉醇(DTX)/依克立达(ELC)白蛋白纳米粒(DTX/ELC Co-loaded NPs),并评价其理化性质。方法采用超高压微射流技术制备DTX/ELC Co-loaded NPs;首先以三氯甲烷-无水乙醇配比、油相-水相配、白蛋白浓度、pH值、微射流压力、微射流循环次数、有机溶剂挥发时间作为考察的处方和工艺变量,以白蛋白纳米粒的粒径分布作为评价指标,通过Plackett-Burman实验设计筛选出对DTX/ELC Co-loaded NPs制剂性质影响的关键性变量;再利用Box-Behnken实验设计优化关键性变量,得到最佳处方工艺。采用动态激光散射仪测定白蛋白纳米粒的粒径分布和Zeta电位;超滤离心法测定纳米粒中两种药物的包封率,并用透射电镜观察其微观结构;通过动态反向膜透析法考察DTX/ELC Co-loaded NPs在磷酸盐缓冲液(pH7.4、pH6.0)中的药物释放特性。结果白蛋白浓度、微射流压力和微射流循环次数是影响DTX/ELC Co-loaded NPs粒径大小的关键性变量。经Box-Behnken实验设计优化得到白蛋白纳米粒的最佳处方工艺为:白蛋白浓度为60 mg·mL^(-1),微射流压力为1.4×10^(4)psi,微射流循环次数为6次。根据最优处方工艺制备3批纳米粒的平均粒径为247.6±15.8 nm,Zeta电位为-22.1±1.1 mV,DTX、ELC的包封率分别为95.6%±1.3%、97.5%±2.3%。DTX/ELC Co-loaded NPs中两种药物在药物释放前期均较快,后期较为平缓,具有缓释效果。结论所用处方设计科学合理,工艺可行,可规模化生产。

OBJECTIVE To prepare Docetaxel and Elacridar albumin nanoparticles( DTX/ELC Co-loaded NPs) and to evaluate their physical and chemical properties. METHODS DTX/ELC Co-loaded NPs were prepared by ultra-high pressure microfluidization technology. The variables of formulation and process were investigated based on chloroform/anhydrous ethanol ratio,oil/water phase ratio,albumin concentration,pH value,microfluidization pressure,microfluidization times,organic solvent volatilization time,and particle size distribution of albumin nanoparticles as the evaluation index. The formulation properties of DTX/ELC Co-loaded NPs were screened through the Plackett-Burman experiment design. Then,Box-Behnken experiment design was used to optimize key variables and obtain the best formulation and process of DTX/ELC Co-loaded NPs. The particle size distribution and Zeta potential of DTX/ELC Co-loaded NPs was measured using a dynamic laser scatterometer. The encapsulation efficiency of Docetaxel and Elacridar in albumin nanoparticles was measured by ultrafiltration centrifugation. The microstructure of DTX/ELC Co-loaded NPs was observed under transmission electron microscopy. Drug release characteristics of DTX/ELC Co-loaded NPs in phosphate buffer( pH 7. 4 and pH 6. 0) were compared by reverse dialysis. RESULTS The Plackett-Burman screening experiment showed that albumin concentration,microfluidization pressure and microfluidization times were the key variables. The optimal formulation and process of DTX/ELC Co-loaded NPs was optimized as follows: albumin concentration was60 mg·mL^(-1),microfluidization pressure was 1. 4 × 10^(4) psi;and microfluidization times was 6 times. The three batches of DTX/ELC Co-loaded NPs were prepared according to the optimal formulation and process. The average particle size was 247. 6 ± 15. 8 nm,and the Zeta potential was-22. 1 ± 1. 1 m V. Encapsulation efficiency of Docetaxel and Elacridar were 95. 6% ± 1. 3% and 97. 5% ± 2. 3%. The drug release of DTX/ELC Co-loaded NPs were faster in the early stage,and slower in the later stage.CONCLUSION The formulation was scientific and reasonable,and the process was feasible,which could be produced on a large scale.