目的 制备转铁蛋白受体结合肽HAIYPRH修饰的负载鬼臼毒素脂质体（HAIYPRH modified podophyllotoxin-loaded liposomes，T7-Pod-Lip），对其处方工艺进行优化，并初步评估其靶向能力。方法 采用薄膜分散法制备T7-Pod-Lip。通过Box-Behnken设计-响应面法（Box-Behnken design-response surface method，BBD-RSM）确定T7-Pod-Lip的最优处方及制备工艺，并对优化处方工艺后的T7-Pod-Lip的形态、粒径、ζ电位、体外释放、稳定性进行考察；考察小鼠脑内血管内皮细胞（bEnd.3）、大鼠脑胶质瘤细胞（C6）对T7-Pod-Lip的体外摄取能力及体外跨血脑屏障（blood brain barrier，BBB）能力，并利用脑胶质瘤小鼠考察T7-Pod-Lip的体内靶向能力。结果 T7-Pod-Lip的最优处方工艺为EPC 34 mg、胆固醇6 mg、鬼臼毒素1 mg、DSPE-PEG₂₀₀₀ 2 mg、DSPE-PEG₂₀₀₀-HAIYPRH 1 mg、超声功率630 W；其平均粒径为（90.94±0.95）nm，ζ电位为（−4.83±0.61）mV，鬼臼毒素包封率为（96.56±0.44）%，且稳定性较好。HAIYPRH修饰的脂质体在bEnd.3、C6细胞中的摄取能力显著增强，且体外跨BBB能力显著增强；活体成像结果表明，HAIYPRH修饰的脂质体在小鼠脑内的荧光信号显著增强，说明具有较好的脑靶向能力。结论 成功制备并优化T7-Pod-Lip，并证明经HAIYPRH修饰的Pod-Lip具有较强的脑靶向作用，为脑胶质瘤的治疗提供了新方法。

Objective To prepare transferrin receptor-binding peptide HAIYPRH modified podophyllotoxin-loaded liposomes (T7-Pod-Lip), optimize formulation process and preliminary evaluate the targeting capability. Methods T7-Pod-Lip was prepared by thin film dispersion method. The optimum preparation process of T7-Pod-Lip was determined by Box-Behnken design-response surface method, and the morphology, particle size, ζ potential, in vitro release and stability of the optimized T7-Pod-Lip were investigated. In vitro uptake and crossing blood brain barrier (BBB) capacities of T7-Pod-Lip by cerebral vascular endothelial cells (bEnd.3) and cerebral glioma cells (C6) were investigated, and in vivo targeting ability of T7-Pod-Lip was investigated by glioma mice. Results The optimal prescriptions of T7-Pod-Lip were EPC 34 mg, cholesterol 6 mg, podophyllotoxin 1 mg, DPE-PEG₂₀₀₀ 2 mg, DSPE-PEG₂₀₀₀-HAIYPRH 1 mg, ultrasonic power 630 W. Its particle size was (90.94 ±0.95) nm, ζ potential was (−4.83 ±0.61) mV, podophyllotoxin encapsulation rate was (96.56 ±0.44)%, and the stability was perfect. The uptake capacity of HAIYPRH modified liposomes in bEnd.3 and C6 cells was significantly enhanced, and the trans BBB capacity was significantly enhanced in vitro. In vivo imaging results showed that the fluorescence signal of HAIYPRH modified liposomes in mouse brain was significantly enhanced, indicating that it had ideal brain targeting ability. Conclusion T7-Pod-Lip is successfully prepared and optimized, and it is proved that Pod-Lip modified by HAIYPRH has strong targeting effect, which provides a new method for the treatment of brain glioma.

R283.6

国家自然科学基金资助项目（82204629）;辽宁省教育厅科学技术研究重点攻关项目（202064）