电荷翻转型聚苹果酸纳米接枝物的制备与性质研究

Nanoconjugate of 2,3-Dimethylmaleic Anhydride-Decorated Polyethyleneimine-Poly( β-L-Malic Acid )-Doxorubicin: Preparation,Characterization and Biological Evalution

目的制备p H-敏感、电荷翻转型聚苹果酸纳米接枝物(多柔比星聚苹果酸-聚乙烯亚胺-2,3-二甲基马来酸酐,DOX/PMLA-PEI-DMA),研究其电荷翻转特性、体外释药、细胞内摄作用以及体外细胞毒性。方法以L-天冬氨酸为原料合成β-聚苹果酸,将聚乙烯亚胺、多柔比星以酰胺键连接到β-聚苹果酸骨架上,电荷保护基团2,3-二甲基马来酸酐通过羟基乙胺连接到β-聚苹果酸上,制备得到纳米接枝物多柔比星聚苹果酸-聚乙烯亚胺-2,3-二甲基马来酸酐。核磁共振光谱表征β-聚苹果酸及纳米接枝物的结构,动态透析法模拟体外释药特性,贝克曼粒度分析仪测定不同p H条件下纳米接枝物的Zeta电位,采用Huh7细胞研究纳米接枝物的内摄作用和细胞毒性。结果成功制备药物载体β-聚苹果酸和纳米接枝物多柔比星聚苹果酸-聚乙烯亚胺-2,3-二甲基马来酸酐;证实多柔比星聚苹果酸-聚乙烯亚胺-2,3-二甲基马来酸酐具有p H-敏感、电荷翻转特性,且其药物的释放具有p H依赖性;多柔比星聚苹果酸-聚乙烯亚胺-2,3-二甲基马来酸酐能够有效地提高多柔比星的细胞内摄作用,细胞毒性实验证实其具有很好的抗肿瘤能力。结论纳米接枝物多柔比星聚苹果酸-聚乙烯亚胺-2,3-二甲基马来酸酐是一种潜在的p H响应型靶向给药体系。

OBJECTIVE To synthesize the biomacromolecule of poly-malic acid（ PMLA） for preparation of a novel 2,3-dimethylmaleic anhydride-decorated polyethyleneimine-poly（ β-L-malic acid）-doxorubicin nanoconjugate for effective and specific drug delivery. METHODS The structures of PMLA and the nanoconjugate were confirmed by1H-NMR. Then the conjugation efficiency and drug release property were determined. The cellular uptake and cytotoxicity were assessed by using human hepatocellular carcinoma（ HCC）cell line Huh7 as in vitro cell model. RESULTS PMLA and DOX / PMLA-PEI-DMA were successfully prepared. The nanoconjugate possessed p H-sensitive charge-conversion and p H-dependent drug release properties. DOX / PMLA-PEI-DMA enhanced the cellular uptake of DOX and in vitro cytotoxicity effectively. CONCLUSION Nanoconjugate DOX / PMLA-PEI-DMA can be used as a promising drug carrier for its targeted and intracellular delivery.