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
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摘要
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 (r_e),target efficacy (t_e) and peak concentration ratio (C_e) were used to evaluate the brain targeting ability of Tf-NefNL. Results: Compared with that of NefNL,C_(max) and AUC of Tf-NefNL increased significantly,T_(max) was delayed notably and t_(1/2) 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.
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 (r_e),target efficacy (t_e) and peak concentration ratio (C_e) were used to evaluate the brain targeting ability of Tf-NefNL. Results: Compared with that of NefNL,C_(max) and AUC of Tf-NefNL increased significantly,T_(max) was delayed notably and t_(1/2) 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.
引文
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11 Ishida O,Maruyama K,Tanahashi H,et al.Liposomes bearing polyethyleneglycol-coupled transferrin with intracellular targeting property to the solid tumors in vivo[J].Pharm Res,2001,18(7):1042-1048
12 Anabousi S,Bakowsky U,Schneider M,et al.In vitro assessment of transferrin-conjugated liposomes as drug delivery systems for inhalation therapy of lung cancer[J].Eur J Pharm Sci,2006,29(5):367-374
13方亮.药剂学[M].第8版.北京:人民卫生出版社,2016:349
2张会杰,熊玉卿.甲基莲心碱小鼠脑内分布的研究[J].中国药师,2010,13(1):3-5
3熊玉卿,曾繁典.甲基莲心碱对有机磷中毒小鼠胆碱酯酶的重活化作用[J].中国药理学通报,2001,17(6):672-675
4宋宁,李颖寰.拉莫三嗪纳米脂质体的制备及脑靶向作用的研究[J].首都医科大学学报,2017,38(1):116-121
5黄静,何文,李秀芳,等.天麻素纳米脂质体的研制及其脑靶向性的初步研究[J].广东药学院学报,2011,27(4):345-350
6蒋新国.脑靶向递药系统的研究进展[J].复旦学报(医学版),2012,39(5):441-448
7王军,谈弋.甲基莲心碱纳米脂质体在小鼠体内组织分布的研究[J].中国药师,2011,14(5):612-614
8 Santos ND,Allen C,Doppen AM,et al.Influence of poly(ethylene glycol)grafting density and polymer length on liposomes:relating plasma circulation lifetimes to protein binding[J].Biochim Biophys Acta,2007,1768(6):1367-1377
9 Lv Q,Li LM,Han M,et al.Characteristics of sequential targeting of brain glioma for transferrin-modified cisplatin liposome[J].Int JPharm,2013,444(1-2):1-9
10谈弋,何文,杨淑清.甲基莲心碱纳米脂质体的研制及其体外释药特性考察[J].广东药学院学报,2010,26(3):226-229
11 Ishida O,Maruyama K,Tanahashi H,et al.Liposomes bearing polyethyleneglycol-coupled transferrin with intracellular targeting property to the solid tumors in vivo[J].Pharm Res,2001,18(7):1042-1048
12 Anabousi S,Bakowsky U,Schneider M,et al.In vitro assessment of transferrin-conjugated liposomes as drug delivery systems for inhalation therapy of lung cancer[J].Eur J Pharm Sci,2006,29(5):367-374
13方亮.药剂学[M].第8版.北京:人民卫生出版社,2016:349