共载姜黄素与阿霉素胶束的制备工艺优化及体外抗肿瘤评价

Optimization of preparation and in vitro anti-tumor evaluation of CUR/DOX micelles

目的 制备共载姜黄素/阿霉素聚己内酯-聚乙二醇-聚己内酯胶束(CUR/DOX micelle),优化其制备工艺,并进行体外抗肿瘤评价。方法 采用薄膜-水化-超声法制备CUR/DOX micelle,以粒径、包封率和载药量为评价指标,采用正交设计优化纳米粒处方;对胶束进行表征,透射电镜观察胶束的外观形态,测定其粒径大小及分布,并测定包封率及载药量;考察其稳定性和体外释药特性;体外抗肿瘤评价中,流式细胞仪考察CUR/DOX micelle的细胞摄取率,MTT法考察其对肿瘤细胞的抑制作用。结果 CUR/DOX micelle的最佳制备工艺为:药物和聚己内酯-聚乙二醇-聚己内酯(PCL-PEG-PCL)比例为1∶10,水化温度为65℃,超声时间为6 min;制得的胶束平均粒径为150 nm,TEM结果显示胶束呈球形;姜黄素的包封率为85.21%,载药量为6.80%,阿霉素的包封率为82.92%,载药量为1.28%;CUR/DOX micelle具有良好的稳定性和缓释能力;体外细胞实验结果显示,CUR/DOX micelle细胞摄取率升高,能有效抑制人乳腺癌MCF-7细胞的增殖。结论 CUR/DOX micelle制备工艺简单且稳定性好,能有效增强药物的体外抗肿瘤活性。

Objective To prepare curcumin and doxorubicin loaded PCL-PEG-PCL micelle(CUR/DOX micelles), optimize the preparation, and evaluate the anti-tumor activity of CUR/DOX micelles in vitro. Methods The CUR/DOX micelles were prepared by thin film-hydration-ultrasound method.The preparation technology was optimized by orthogonal test with the particle size, encapsulation efficiency and drug loading as the evaluation parameters. The morphology imaging of CUR/DOX micelles was observed by transmission electron microscopy. The particle size was conducted by a nanoparticle analyzer. The drug-loading rate and encapsulation efficiency of micelle were determined.The stability and release profile in vitro were determined. The cellular uptake of CUR/DOX micelles was measured by flow cytometry and the cytotoxicity was determined by MTT in vitro. Results The optimal preparation of CUR/DOX micelles was as follows: the drug and PCL-PEG-PCL ratio was 1∶10, the hydration temperature was 65 ℃, and the ultrasound use for 6 min. The optimal mean diameter of micelles was around 150 nm at size and the TEM showed spherical morphology. The encapsulation efficiency and drug loading of curcumin were 85.21% and 6.80%;those of doxorubicin were 82.92% and 1.28%, respectively. The CUR/DOX micelles exhibited good stability and sustained release. In vitro experiments showed CUR/DOX micelles increased the cellular uptake efficiency and enhanced the cytotoxicity in MCF-7 cells effectively. Conclusion The preparation method is simple and stable, and CUR/DOX micelles can enhance the anti-tumor activity significantly.