目的 制備聚乙二醇（PEG）和八聚精氨酸（R8）雙修飾漆黃素脂質(zhì)體（PEG and R8 co-modified fisetin liposomes，PEG/R8-Fis-Lips），對(duì)其進(jìn)行理化表征，并考察其口服相對(duì)生物利用度和改善急性肝損傷作用。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）并進(jìn)行核磁共振氫譜（¹H-NMR）確認(rèn)。采用后插入法制備PEG/R8-Fis-Lips；HPLC法測(cè)定漆黃素含量并計(jì)算包封率及載藥量；單因素考察PEG/R8-Fis-Lips處方工藝，采用Box-Behnken設(shè)計(jì)-響應(yīng)面法（Box-Behnken design-response surface method，BBD-RSM）優(yōu)化PEG/R8-Fis-Lips處方，并采用乳糖將PEG/R8-Fis-Lips混懸液制備成凍干粉；透射電子顯微鏡（transmission electron microscopy，TEM）觀察其形態(tài)，X射線粉末衍射法考察凍干粉晶型，考察PEG/R8-Fis-Lips凍干粉在模擬消化液中的穩(wěn)定性、體外釋藥行為及貯存穩(wěn)定性。SD大鼠ig給予PEG/R8-Fis-Lips后采血，考察其口服藥動(dòng)學(xué)行為；建立急性肝損傷模型，考察PEG/R8-Fis-Lips改善急性肝損傷作用。結(jié)果 成功合成了DSPE-mPEG₂₀₀₀-R8。PEG/R8-Fis-Lips最佳處方：磷脂與膽固醇用量比為6.0∶1，總脂質(zhì)與藥物用量比為12.5∶1，DSPE-mPEG₂₀₀₀-R8質(zhì)量濃度為0.26 mg/mL；PEG/R8-Fis-Lips的包封率、載藥量、粒徑和ζ電位分別為（86.17±0.20）%、（6.01±0.10）%、（253.75±13.14）nm、（−14.16±0.82）mV，PEG/R8-Fis-Lips外觀為球形及類球形。漆黃素在PEG/R8-Fis-Lips凍干粉中以無(wú)定型狀態(tài)存在，PEG/R8-Fis-Lips在模擬消化液中的穩(wěn)定性及累積釋放率均高于漆黃素原料藥及其普通脂質(zhì)體（Fis-Lips），其體外釋藥過(guò)程符合Weibull模型；貯存穩(wěn)定性也明顯提高。藥動(dòng)學(xué)結(jié)果顯示，PEG/R8-Fis-Lips達(dá)峰濃度（C_max）增加至（604.05±166.73）ng/mL，半衰期（t_1/2）延長(zhǎng)至（5.04±0.63）h，相對(duì)生物利用度提高至7.71倍；且PEG/R8-Fis-Lips減輕了對(duì)乙酰氨基酚所致的急性肝損傷。結(jié)論 PEG/R8-Fis-Lips極大地促進(jìn)了漆黃素口服吸收，并增強(qiáng)了漆黃素改善急性肝損傷作用。

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

國(guó)家自然科學(xué)基金項(xiàng)目（81670088）;鄭州澍青醫(yī)學(xué)高等?？茖W(xué)校教學(xué)創(chuàng)新團(tuán)隊(duì)項(xiàng)目（2024jxcxtd01）