具有肿瘤靶向及穿膜效应的壳聚糖胶束的制备

Preparation of tumor-targeting chitosan micelle with transmembrane effect

在壳聚糖(CS)表面修饰疏水基团辛基形成辛基壳聚糖(OC),然后再修饰亲水基团聚乙二醇(PEG)、肿瘤靶向配体氨基葡萄糖(DG)和穿膜肽九聚精氨酸(9R),形成DG和9R共同修饰的、同时具有肿瘤靶向性及穿膜效应的壳聚糖纳米胶束(DG/9R-PEG-OC)。核磁共振光谱分析及聚丙烯酰胺凝胶电泳检测结果证实了DG/9R-PEG-OC的成功制备;测得壳聚糖纳米胶束的粒径为151.8 nm左右、Zeta电位约为16.5 m V;透射电子显微镜照片显示该壳聚糖纳米胶束为均匀的球形结构;紫外分光光度法测定该载体的荧光素载药量约为28.2%,包封率约为75.0%,释药实验表明壳聚糖胶束具有良好的缓释作用;荧光显微镜观察显示,该DG/9R-PEG-OC胶束对肿瘤细胞尤其是葡萄糖受体高表达的肿瘤细胞Hep G2具有较好的靶向性及细胞穿膜效应。故DG/9R-PEG-OC胶束可作为脂溶性抗肿瘤药物的载体用于肿瘤的靶向化学治疗。

Chitosan（CS）surface was modified with hydrophobic octyl groups to prepare N-octyl chitosan（nitro-gen-octyl chitosan;OC）.Then hydrophilic group carboxyl-polyethylene glycol-amino （PEG）;tumor-targeting lig-and D-glucosamine（DG）;and membrane-penetrating peptide 9-D-arginine（9R）were linked to OC successively. Then the DG and 9R modified chitosan micelle （DG/9R-PEG-OC）with tumor-targeting and transmembrane effect was prepared.By hydrogen nuclear magnetic resonance spectrometer （1 H NMR）and sodium dodecyl sul-fate polyacryl amide gel electrophoresis（SDS-PAGE）;the successful formation of DG/9R-PEG-OC was certified;with particle size of 151.8 nm and Zeta potential of 16.5 mV.The morphology of chitosan micelle observed by transmission electron microscope was homogeneous spherical structure.The drug loading content （DLC）（using fluorescein as a model drug）and encapsulation efficiency （EE）were about 28.2% and 75.0% measured by UV-visible spectrophotometer.Meanwhile;the drug showed a controlled releasing profile out of the micelle.Cellu-lar uptake experiments indicated DG/9 R-PEG-OC micelle had a significant tumor-tageting and transmembrane effects;especially on HepG2 cells;which exbihited high expression of the glucose transporter.Thus DG/9R-PEG-OC micelle could be a promising drug targeted delivery system of hydrophobic antitumor drugs.