转铁蛋白修饰的甲基莲心碱纳米脂质体的研制及脑靶向性考察

Preparation and Brain Targeting Ability of Transferrin-modified Neferine Nanolipoosmes

目的:制备转铁蛋白(Tf)修饰的甲基莲心碱(Nef)纳米脂质体(NefNL),并考察其脑部靶向性。方法:以二硬脂酰基磷脂酰胆碱(DSPC)、胆固醇、二硬脂酰磷脂酰乙醇胺-聚乙二醇2000(DSPE-PEG2000)及二硬脂酰磷脂酰乙醇胺-聚乙二醇2000-羧基(DSPE-PEG200-COOH)为成膜材料,采用薄膜分散法制备NefNL,并将Tf共价结合于DSPE-PEG200-COOH的COO-上制备Tf修饰的NefNL(Tf-NefNL)。HPLC法测定小鼠中血浆及脑部Nef浓度。以同剂量的NefNL为对照,测定Tf-NefNL给药后不同时间点小鼠血浆及脑部的Nef浓度,并利用DAS2.0软件计算主要药动学参数,并用相对摄取率(r_e)、靶向效率(t_e)及峰浓度比(C_e) 3个指标评价其脑靶向性。结果:与同剂量的NefNL相比,Tf-NefNL在小鼠脑组织中的峰浓度(C_(max))及曲线下面积(AUC)明显增加,达峰时间(T_(max))缩短(P<0.05),r_e、t_e、C_e分别为1.90、1.37、2.0。结论:Tf修饰的纳米脂质体可显著增加Nef在小鼠脑部的浓度,加快达峰,具有明显的脑靶向性。

Objective: To prepare transferrin (Tf)-modified neferine (Nef) nanoliposomes (NefNL) and explore the brain targeting ability. Methods: Using distearyl phosphatidyl cholin (DSPC),cholesterol,distearoyl phosphoethanolamine-polyethylene glycol2000 (DSPE-PEG2000) and DSPE-PEG200-COOH as the membrane materials,NefNL was prepared by a film-dispersion method. Tfmodified NefNL (Tf-NefNL) was then obtained by the covalent bonding of Tf with the terminal COO-of DSPE-PEG200-COOH. The contents of Nef in plasma and brain of mice were determined by HPLC. NefNL at the same dose was used as the positive control,and the contents of Nef in plasma and brain of mice were detected at various predetermined time points after the injection of Tf-NefNL. DAS2.0 software was used to calculate the main pharmacokinetic parameters. Three indices including relative intake rate (re),target efficacy (te) and peak concentration ratio (Ce) were used to evaluate the brain targeting ability of Tf-NefNL. Results: Compared with that of NefNL,Cmax and AUC of Tf-NefNL increased significantly,Tmax was delayed notably and t1/2 in plasma was prolonged obviously as well (P<0.05). The three target indices of re,teand Cewere 1.90,1.37 and 2.0. Conclusion: Tf-modified nanoliposomes can increase brain distribution of Nef in mice with quicker effective time,suggesting notable brain targeting ability.