载多烯紫杉醇靶向脂质微泡的制备及其体外寻靶能力

Preparation and targeting ability in vitro of DR5-targeted lipid microbubble loaded with docetaxel

目的制备一种携带肝癌DR5单克隆抗体(DR5mAb)的载多烯紫杉醇靶向脂质超声微泡(Docetaxel-loaded lipidmicrobubbles,DLLM),并检测其体外寻靶能力。方法采用机械振荡法制备载多烯紫杉醇的DLLM,并制备生物素化的肝癌DR5mAb,通过生物素-亲和素连接方法,将DR5mAb连接于载多烯紫杉醇的DLLM表面,制备DR5-DLLM。检测DR5-DLLM的粒径、浓度、Zeta电位、包封率、载药量、稳定性及其体外寻靶能力。结果 DR5-DLLM的平均粒径为1 232 nm,Zeta电位为-9.86 mV,平均浓度为3.1×109个/ml,微泡的包封率为73.5%,平均载药量为25.3%。60Co射线灭菌前后以及4℃、-20℃保存14 d,微泡的形态、包封率、载药量未见明显改变。靶向微泡组的HepG2细胞周围可见DR5-DLLM微泡紧密结合,而非靶向微泡DLLM组未见特异性微泡结合。结论成功制备了携带肝癌DR5mAb的载多烯紫杉醇靶向脂质超声微泡,该微泡能够与HepG2细胞牢固结合,有望成为一种新型、高效、可用于超声分子显像的肝癌靶向药物载体。

Objective To prepare the docetaxel-loaded lipid microbubbles （DLLM） coupled with DR5mAb against liver cancer, and determine its targeting ability in vitro. Methods DLLM were prepared by mechanical oscillation, based on which DR5- targeted docetaxel-loaded lipid microbubbles （DR5-DLLM） were prepared by attaching biotinylated DR5mAb to the surface of DLLM via avidin-biotin interaction, and determined for size, concentration, Zeta potential, entrapment efficiency, drug-load, stability and targeting ability in vitro. Results The mean size, Zeta potential, mean concentration, entrapment efficiency and mean drug-load of DR5,DLLM were 1 232 rim, -9. 86 mV, 3. 1 ×109 microbubbles/ml, 73. 5% and 25. 3%, respectively. No obvious change was observed in appearance, entrapment efficiency or drug-load of DRS-DLLM before and after sterilization with ℃o and after storage at 4 and -20℃ for 14 d. DR5-DLLM were tightly conjugated with HepG2 cells, while DLLM showed no specific conjugation with the cells. Conclusion DR5-DLLM was successfully prepared, which were conjugated with HepG2 cells tightly, and might be used as a novel and highly effective liver cancer-targeted drug carrier for uhrasonund molecular imaging.