双修饰眼镜蛇神经毒素脂质体的制备及其经鼻黏膜给药后在大鼠脑内药动学研究  

Preparation of Co-modified Liposomes of α-Cobrotoxin and Its Brain Pharmacokinetics Study in Rats after Intranasal Administration

作　　者：朱潇冉 柳琳[2] 钱丽梅[1] 杨慧[1] 张幸国[2] 

机构地区：[1]浙江中医药大学药学院,杭州310053 [2]浙江大学医学院附属第一医院药剂科,杭州310003

出　　处：《中国药学杂志》2016年第10期815-820,共6页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(81503256/H2806);浙江省中医药科学研究基金资助项目(2015ZA046)

摘　　要：目的研究双修饰眼镜蛇神经毒素脂质体[(Pep2(Pep1)-αCT-LP)]鼻黏膜给药后药物在大鼠脑中的药动学行为。方法通过Michael加成反应合成二硬脂酰甘油磷脂乙醇胺-聚乙二醇-Pep1及二硬脂酰甘油磷脂乙醇胺-聚乙二醇-Pep2,通过核磁共振氢谱(~1H-NMR)和红外光谱(FTIR)验证产物结构。采用薄膜分散-后插入法制备双修饰眼镜蛇神经毒素脂质体,制得的脂质体考察形态、粒径、Zeta电位;采用超滤离心法测定脂质体包封率。应用大鼠脑微透析技术测定鼻黏膜给药后大鼠脑中枢中脑导水管周围灰质(periaqueductal gray,PAG)部位未修饰、单修饰以及双修饰眼睛蛇神经毒素脂质体药物浓度经时变化,用PKSolver软件处理药动学参数。结果核磁共振氢谱和红外光谱证实二硬脂酰甘油磷脂乙醇胺-聚乙二醇-Pep1及二硬脂酰甘油磷脂乙醇胺-聚乙二醇-Pep2结构。制备的双修饰眼镜蛇神经毒素脂质体为球形或近球形,大小均一,粒径较小(115.8±1.86)nm,Zeta电位为(-13.77±0.75)m V,包封率为(32.75±1.12)%。在中脑导水管周围灰质部位,双修饰眼镜蛇神经毒素脂质体最大药物浓度(ρ_(max))显著高于其他各组脂质体(P<0.05),ρ_(max)、t_(max)、AUC_(0→∞)分别为(244.72±3.15)ng·mL^(-1)、(88.01±4.19)min、(89 199.02±1 922.99)ng·min·mL^(-1)。结论双修饰眼镜蛇神经毒素脂质体可以显著提高药物在中脑导水管周围灰质部位浓度,该结果为研究蛋白多肽类药物经鼻黏膜给药的脑靶向制剂的开发提供了参考。OBJECTIVE To investigate the brain pharmacokinetics of co-modified liposomes of α-cobrotoxin in rats after intranasal administration. METHODS DSPE-PEG（2000）-Pep1 and DSPE-PEG（2000）-Pep2 were synthesized by Michael addition reaction. And their structures were verified by proton nuclear magnetic resonance spectroscopy（^1H-NMR） and infrared spectroscopy（ FTIR）. Pep2（ Pep1）-αCT-LP was prepared by the method of thin-film hydration and post-inserting,then its morphology,particle size,and Zeta potential were investigated. The encapsulation efficiency of liposomes was determined by ultrafiltration centrifugal method. The concentrations of αCTLP,Pep1-αCT-LP,Pep2-αCT-LP,and Pep2（ Pep1）-αCT-LP in periaqueductal gray（ PAG） after intranasal administration were measured by microdialysis and the pharmacokinetical parameters were analyzed by PKSolver software. RESULTS The structures of DSPEPEG（2000）-Pep1 and DSPE-PEG（2000）-Pep2 were proved by-1H-NMR and FTIR. The prepared Pep2（ Pep1）-αCT-LP was nearly spherical with uniform size,the mean particle size was（ 115. 8 ± 1. 86） nm,and the Zeta potential was（- 13. 77 ± 0. 75） m V. Besides,the encapsulation efficiency was（ 32. 75 ± 1. 12） %. The results of in vivo test demonstrated that the αCT concentrations in PAG after intranasal administration of Pep2（ Pep1）-αCT-LP were significantly increased compared with the groups of αCT-LP,Pep1-αCT-LP and Pep2-αCT-LP（ P〈0. 05）. The ρ（max）,t（max）,and AUC（0→∞） were（ 244. 72 ± 3. 15） ng · mL^-1,（ 88. 01 ± 4. 19） min,（ 89 199. 02 ± 1 922. 99）ng·min·mL^-1,respectively. CONCLUSION Pep2（ Pep1）-αCT-LP can significantly increase the concentrations of αCT in periaqueductal gray,which provides a promising method for development of polypeptide agents for brain-targeting.

关 键 词：眼镜蛇神经毒素 双修饰脂质体 微透析 鼻黏膜给药 

分 类 号：R944[医药卫生—药剂学]