叶酸与端醛基聚乙二醇双重修饰的壳聚糖-硬脂酸纳米胶束的制备

Preparation of Chitosan-stearic Acid Nano-micelles Modified with Folic Acid and mPEG

目的:制备一种包载难溶性抗肿瘤药物的聚合物胶束,以提高难溶性抗肿瘤药物的抑瘤作用。方法:先用壳聚糖(CSO)、硬脂酸(SA)制成胶束(CSO-SA),再依次以端醛基聚乙二醇(mPEG)和叶酸(FA)对其进行修饰制成胶束(PEG-CSO-SA和FA-PEG-CSO-SA),采用红外光谱检测CSO-SA、PEG-CSO-SA、FA-PEG-CSO-SA的特征官能团,透射电镜观察胶束的微观形态,激光粒度测定仪测定胶束的粒径和Zeta电位。以蛇床子素(OST)为模型药,采用透析法制备载药纳米胶束(FA-PEG-CSO-SA/OST),以MTT法检测FA-PEG-CSO-SA、OST溶液和FA-PEG-CSO-SA/OST对人肝癌细胞HepG2的抑制率,并计算半数抑制浓度(IC50)。结果:成功制得FA-PEG-CSO-SA。CSO-SA、PEG-CSO-SA、FA-PEG-CSO-SA均呈椭圆形,粒径分别为(96.01±5.99)、(112.93±1.06)、(216.01±4.76)nm(n=3),Zeta电位分别为(39.30±1.75)、(38.03±2.91)、(15.17±2.10)mV(n=3)。FA-PEG-CSO-SA/OST中OST的包封率为(84.47±2.07)%,载药量为(16.01±0.90)%(n=3),FA-PEG-CSO-SA对HepG2细胞的抑制率<20%,OST溶液和FA-PEG-CSO-SA/OST对HepG2细胞的IC50分别为(62.08±5.21)、(27.49±0.50)μg/mL(n=3)。结论:所制FA-PEG-CSO-SA能明显提高难溶性药物OST对HepG2细胞的抑瘤作用,其有望成为一种新型的抗肿瘤药物载体。

OBJECTIVE:To prepare insoluble anti-tumor drug-loading polymer micelles,and to increase inhibitive effect of insoluble anti-tumor drug.METHODS:Chitosan(CSO)and stearic acid(SA)were used to prepare blank micelles(CSO-SA),then modified with mPEG and folic acid(FA)to prepare PEG-CSO-SA and FA-PEG-CSO-SA.Characteristic functional groups of CSO-SA,PEG-CSO-SA and FA-PEG-CSO-SA were detected by infrared spectroscopy.The morphology of micelles was observed by transmission electron microscopy.The particle size and Zeta potential of micelles were measured by laser particle size analyzer.Osthole(OST)was used as the model drug and drug-loading micelles(FA-PEG-CSO-SA/OST)were prepared by dialysis.MTT assay was used to detect the inhibitory rate of FA-PEG-CSO-SA,OST solution and FA-PEG-CSO-SA/OST to human liver cancer cell HepG2.Half inhibitory concentration(IC50)was calculated.RESULTS:FA-PEG-CSO-SA was successfully prepared.CSO-SA,PEG-CSO-SA,FA-PEG-CSO-SA were oval in shape;particle sizes were(96.01±5.99),(112.93±1.06),(216.01±4.76)nm(n=3)and Zeta potentials were(39.30±1.75),(38.03±2.91),(15.17±2.10)mV(n=3),respectively.Encapsulation efficiency and drug-loading amount of OST in FA-PEG-CSO-SA were(84.47±2.07)%and(16.01±0.90)%(n=3),respectively.The inhibition rates of FA-PEG-CSO-SA to HepG2 cells were<20%.IC50 of OST solution and FA-PEG-CSO-SA/OST to HepG2 cells were(62.08±5.21),(27.49±0.50)μg/mL(n=3),respectively.CONCLUSIONS:Prepared FA-PEG-CSO-SA can significantly increase inhibitive effect of insoluble drug OST to HepG2 cells,and it is expected to become a new anti-tumor drug carrier.