目的 制备聚乙二醇（PEG）和八聚精氨酸（R8）双修饰漆黄素脂质体（PEG and R8 co-modified fisetin liposomes，PEG/R8-Fis-Lips），对其进行理化表征，并考察其口服相对生物利用度和改善急性肝损伤作用。方法 合成二硬脂酰磷脂酰乙醇胺-聚乙二醇2000-八聚精氨酸（DSPE-mPEG₂₀₀₀-R8）并进行核磁共振氢谱（¹H-NMR）确认。采用后插入法制备PEG/R8-Fis-Lips；HPLC法测定漆黄素含量并计算包封率及载药量；单因素考察PEG/R8-Fis-Lips处方工艺，采用Box-Behnken设计-响应面法（Box-Behnken design-response surface method，BBD-RSM）优化PEG/R8-Fis-Lips处方，并采用乳糖将PEG/R8-Fis-Lips混悬液制备成冻干粉；透射电子显微镜（transmission electron microscopy，TEM）观察其形态，X射线粉末衍射法考察冻干粉晶型，考察PEG/R8-Fis-Lips冻干粉在模拟消化液中的稳定性、体外释药行为及贮存稳定性。SD大鼠ig给予PEG/R8-Fis-Lips后采血，考察其口服药动学行为；建立急性肝损伤模型，考察PEG/R8-Fis-Lips改善急性肝损伤作用。结果 成功合成了DSPE-mPEG₂₀₀₀-R8。PEG/R8-Fis-Lips最佳处方：磷脂与胆固醇用量比为6.0∶1，总脂质与药物用量比为12.5∶1，DSPE-mPEG₂₀₀₀-R8质量浓度为0.26 mg/mL；PEG/R8-Fis-Lips的包封率、载药量、粒径和ζ电位分别为（86.17±0.20）%、（6.01±0.10）%、（253.75±13.14）nm、（−14.16±0.82）mV，PEG/R8-Fis-Lips外观为球形及类球形。漆黄素在PEG/R8-Fis-Lips冻干粉中以无定型状态存在，PEG/R8-Fis-Lips在模拟消化液中的稳定性及累积释放率均高于漆黄素原料药及其普通脂质体（Fis-Lips），其体外释药过程符合Weibull模型；贮存稳定性也明显提高。药动学结果显示，PEG/R8-Fis-Lips达峰浓度（C_max）增加至（604.05±166.73）ng/mL，半衰期（t_1/2）延长至（5.04±0.63）h，相对生物利用度提高至7.71倍；且PEG/R8-Fis-Lips减轻了对乙酰氨基酚所致的急性肝损伤。结论 PEG/R8-Fis-Lips极大地促进了漆黄素口服吸收，并增强了漆黄素改善急性肝损伤作用。

Objective To prepare PEG and R8 co-modified fisetin liposomes (PEG/R8-Fis-Lips), and to investigate its relative bioavailability and protective effects on acute liver injury in SD rats after oral administration. Methods DSPE-mPEG₂₀₀₀-R8 was synthesized and confirmed by ¹H-NMR method. Post-insertion method was used to prepare PEG/R8-Fis-Lips, HPLC method was used to determine the content of fisetin and calculated the encapsulation efficiency and drug loading. Single factor experiments were used to study the prescription process of PEG/R8-Fis-Lips, Box-Behnken design-response surface method (BBD-RSM) was employed to investigate the optimal prescriptions of PEG/R8-Fis-Lips, and its lyophilized powder was prepared using lactose. Morphology of PEG/R8-Fis-Lips was observed by transmission electron microscopy (TEM), crystal form of its lyophilized powder was analyzed by X-ray powder diffraction (XRPD). Stability of PEG/R8-Fis-Lips lyophilized powder in simulated digestive fluid, in vitro drug release behavior and storage stability were also examined, respectively. SD rats were orally administered of PEG/R8-Fis-Lips, and pharmacokinetic behavior was also investigated. The model of acute liver injury was established and then investigated the protective effects of PEG/R8-Fis-Lips on acute liver injury. Results DSPE-mPEG2000-R8 was synthesized successfully. Optimal prescriptions of PEG/R8-Fis-Lips: phospholipids-cholesterol ratio was 6.0:1, total lipids-drug ratio was 12.5:1 and the concentration of DSPE-MPEG₂₀₀₀-R8 was 0.26 mg/mL. Envelopment efficiency, drug loading, particle size and ζ potential of PEG/R8-Fis-Lips were (86.17 ±0.20)%, (6.01 ±0.10)%, (253.75 ±13.14) nm, and (−14.16 ±0.82) mV, respectively. The appearance of PEG/R8-Fis-Lips were spherical or nearly spherical, fisetin was present in an amorphous state in the PEG/R8-Fis-Lips lyophilized powder. The stability and cumulative release rate of PEG/R8-Fis-Lips in simulated digestive fluid were higher than those of fisetin and conventional liposomes (Fis-Lips). Release process of PEG/R8-Fis-Lips accorded with Weibull model, and its storage stability was also significantly improved. Pharmacokinetic results showed that C_max of PEG/R8-Fis-Lips was enhanced to (604.05 ±166.73) ng/mL, t_1/2 was prolonged to (5.04 ±0.63) h, and relative oral bioavailability of PEG/R8-Fis-Lips was enhanced to 7.71 times. PEG/R8-Fis-Lips alleviated acute liver injury induced by paracetamol. Conclusion PEG/R8-Fis-Lips greatly promoted the oral absorption of fisetin, and enhanced its protective effects on acute liver injury.

R283.6

国家自然科学基金项目（81670088）;郑州澍青医学高等专科学校教学创新团队项目（2024jxcxtd01）