“药辅合一”茶多酚-蜂毒肽纳米复合物的制备及抗肿瘤研究

Objective: To prepare epigallocatechin gallate - melittin nanocomposites (EMN) with synergistic antitumor effects and to investigate their antitumor activities. Methods: Polyepigallocatechin gallate (poly EGCG) was synthesized by the Baeyer reaction, and its structure and relative molecular mass were characterized by ¹H - NMR and MS. The preparation process of EMN was optimized by orthogonal experiments, and the in vitro release of melittin (Mel) under different pH values was investigated. The cellular uptake of fluorescein isothiocyanate (FITC) - labeled Mel and EMN was compared by flow cytometry. The synergistic antitumor effects of poly EGCG and Mel in B16F10 and A549 cells were investigated by the MTT method. Results: The structure of the synthesized poly EGCG was confirmed by ¹H - NMR and MS. The optimal preparation process of EMN optimized by orthogonal experiments was as follows: 0.5 mg/mL poly EGCG solution was added dropwise to an equal volume of 1 mg/mL Mel solution, stirred during the addition, and incubated at room temperature for 30 min. The in vitro release experiment showed that the release of EMN was acid - responsive. The uptake experiment showed that both Mel and EMN could be taken up by tumor cells, and the uptake amounts were comparable. The results of the MTT experiment showed that the combination index (CI) of poly EGCG and Mel was less than 1.0, indicating a synergistic antitumor effect. Conclusion: EMN was prepared by a self - assembly method, with a simple process, an appropriate particle size, and a synergistic antitumor effect, worthy of further research.

目的制备具有协同抗肿瘤作用的茶多酚-蜂毒肽纳米复合物(EMN),并考察其抗肿瘤活性。方法通过拜耳反应合成了多聚表没食子儿茶素没食子酸酯(poly EGCG),~1H-NMR和MS表征其结构和相对分子质量。通过正交试验优化EMN的制备工艺,并考察在不同p H值条件下蜂毒肽(Mel)的体外释放。通过流式细胞仪比较异硫氰酸荧光素(FITC)标记的Mel和EMN的细胞摄取情况。MTT法考察poly EGCG和Mel在B16F10和A549细胞的协同抗肿瘤效应。结果合成的poly EGCG用~1H-NMR和MS确证了结构,正交试验优化的EMN最佳制备工艺为将0.5 mg/m L poly EGCG溶液逐滴加入到1 mg/m L等体积的Mel溶液中,边滴加边搅拌,室温孵育30 min,即得。体外释放实验表明EMN释放具有酸响应性。摄取实验表明Mel和EMN均能被肿瘤细胞摄取,摄取量相当。MTT实验结果表明,poly EGCG和Mel联合使用的协同系数(CI)小于1.0,具有协同抗肿瘤效果。结论采用自组装的方式制备EMN,工艺简单,粒径适宜,且具有协同抗肿瘤的效果,值得深入研究。