目的 优化咖啡酸苯乙酯纳米混悬剂的处方,并考察其体外抑制乳腺癌细胞作用。方法 以泊洛沙姆188、蜂胶作为载体,采用反溶剂沉淀法制备,通过星点设计-效应面法优化最佳制备工艺参数,并考察咖啡酸苯乙酯纳米混悬剂的稳定性、载药量、包封率以及冻干保护剂的筛选,同时考察对4T1乳腺癌细胞的生长抑制作用和细胞摄取情况。结果 咖啡酸苯乙酯纳米混悬剂的最优处方为蜂胶0.88 mg/mL、咖啡酸苯乙酯1.86 mg/mL、泊洛沙姆1881 mg/mL,制备得到咖啡酸苯乙酯纳米混悬剂平均粒径为150 nm左右,包封率98%以上,在各种生理介质中都能够稳定存在; 0.5%的BSA对于咖啡酸苯乙酯纳米混悬剂的冻干保护效果最好。咖啡酸苯乙酯纳米混悬剂对4T1乳腺癌细胞的抑制作用与咖啡酸苯乙酯相比提高了2.95倍。咖啡酸苯乙酯纳米混悬剂被细胞摄取后主要分布在细胞质中,随着给药时间的增长,摄取也逐步提高。结论 所得咖啡酸苯乙酯纳米混悬剂具有较高的包封率、载药量,并显著提高了咖啡酸苯乙酯对于4T1细胞的抑制作用。

Objective To optimize formulation of caffeic acid phenethyl ester nanosuspensions (CAPE-NPs) and evaluate its antitumor activity against breast cancer cell in vitro.Methods Poloxam 188 and propolis were used as carriers, and the formulation was prepares by antisolvent precipitation method. Preparation process parameters were optimized using Box-Behnken design- response surface method. The stability, drug loading capacity, and encapsulation efficiency of CAPE-NPs were investigated, and the lyophilization protective agent was screened. At the same time, the growth inhibitory effect and cellular uptake of 4T1 breast cancer cells of CAPENPs was evaluated by MTT assay in vitro.Results The optimal formulation included 0.88 mg/mL propolis, 1.86 mg/mL caffeic acid phenethyl ester, and 1 mg/mL P188. The mean particle size was about 150 nm, and the encapsulation rate was above 98%. CAPE-NPs could stably exist in various physiological media. 0.5% BSA could play a good protective role in the freeze drying process of CAPENPs. The inhibitory effect of CAPE-NPs against 4T1 breast cancer cells was 2.95 times higher than that of caffeic acid phenethyl ester. After being taken up by cells, it is mainly distributed in the cytoplasm, and the uptake of CAPE-NPs gradually increases with the increase of administration time.Conclusion CAPE-NPs has high encapsulation efficiency and drug loading, and can significantly improves the inhibitory effect of caffeic acid phenethyl ester against 4T1 cells.

R285

中国医学科学院医学与健康创新工程重大协同创新项目(2016-12M-1-012)