大黄酸PEGPCLPEI纳米粒的制备及体外评价(1)
陳丹飞1， 朱永琴1， 张源1， 王林燕2， 魏颖慧2*
(1 浙江中医药大学 附属第一医院， 浙江 杭州 310006；
2 浙江中医药大学 药学院， 浙江 杭州 310053)
[摘要]合成聚乙二醇聚己内酯聚乙烯亚胺(PEGPCLPEI)三嵌段聚合物载体材料，制备包载大黄酸(RH)的PEGPCLPEI纳米粒(PPPRHNPS)，并评价其理化性质和体外生物学特性。通过开环聚合和麦克尔加成反应合成得到PEGPCLPEI聚合物，相对分子质量为95×103，临界胶束浓度为0723 nmol·L-1。包载RH制备得到PPPRHNPS，外观浅黄、乳光明显，透射电镜观察纳米粒分散均匀圆整无团聚，粒径为(1183±36)nm，PDI为(019±008)，Zeta电位为(63±15)mV，包封率为(9364±528)%，载药量为(857±053)%。透析法考察PPPRHNPS体外释药特征，48 h内累积释放率为7592%，释药曲线符合Higuchi模型方程：Q=0121 6t1/2+0069 5(R2=0887 4)，呈缓释特性。选用兔红细胞考察其溶血率，MTT法评价其对HK2细胞的生物安全性，在0～005 mmol·L-1不会引起红细胞溶血和细胞毒性。流式细胞仪考察其摄取效率，PPPRHNPS可被细胞迅速内吞，摄取效率高，30 min内摄取完全，经激光共聚焦显微镜观察，PPPRHNPS能够从溶酶体进入细胞质，具有溶酶体逃逸特性。因此，该研究成功合成PEGPCLPEI聚合物和制备得到PPPRHNPS，粒径分布均匀，包封率及载药量较高，摄取迅速，具有缓释特征、溶酶体逃逸特性、优良的生物安全性，是一种良好研究前景的新型纳米制剂。
, http://www.100md.com
[关键词]大黄酸； 聚乙二醇聚己内酯聚乙烯亚胺； 纳米粒； 体外评价
Preparation and in vitro evaluation of rheinloaded
PEGPCLPEI nanoparticles
CHEN Danfei1， ZHU Yongqin1， ZHANG Yuan1， WANG Linyan2， WEI Yinghui2*
(1. The First Affiliated Hospital of Zhejiang Chinese Medical University， Hangzhou 310006， China；
2College of Pharmaceutical Science， Zhejiang Chinese Medical University， Hangzhou 310053， China)
, 百拇医药
[Abstract]This study was aimed to synthesize the polyethyleneglycolpolycaprolactonepolyethyleneimine (PEGPCLPEI) three block polymer material， prepareRhein (RH)loaded PEGPCLPEI nanoparticles(PPPRHNPS)， and then evaluate their physical and chemical properties and biological characteristics in vitro PEGPCLPEI polymer was obtained by adopting theringopening polymerization and Michael addition reaction， and their physical and chemical properties were analyzed by using NMR and gel permeation chromatography PEGPCLPEI was then used as the carriers to prepare PPPRHNPS by applying spontaneous emulsification solvent diffusion method The results showed that molecular weight of PEGPCLPEI polymer was 95×103， and critical micelle concentration was 0723 mmol·L-1 PPPRHNPS had pale yellow， opalescence faade， round and smooth without aggregation， formed of (1183(36) nm in particle size with PDI of (019±008)， Zeta potential of (63±15) mV， entrapment efficiency of (9364±528)%， and drug loading of (857±053)% The accumulative release percentage of PPPRHNPS was 7592% in 48h， and the release profiles in PBS conformed to the Higuchi equation： Q=0121 6t1/2+0069 5 (R2=0887 4)， presenting slow release characteristics Within the scope of the 0005 mmol·L-1， the nanoparticles had no obvious hemolysis on rabbit red blood cells and toxicity on HK2 cells In the investigation of uptake efficiency by flow cytometry， nanoparticles can be absorbed into cells quickly and internalized within 30 minutes fully， with a high uptake efficiency In confocal laser scanning microscope observation， the nanoparticles can escape from lysosome into cytoplasm Herein， this study synthesized the PEGPCLPEI polymer and prepared PPPRHNPS successfully； the nanoparticles showed uniform particle size， higher encapsulation efficiency and drugloading rate， slow release characteristics， quick uptake and internalization， lysosome escape property and good biocompatibility PPPRHNPS will be a promising pharmaceutical formulation for further development, http://www.100md.com
(1 浙江中医药大学 附属第一医院， 浙江 杭州 310006；
2 浙江中医药大学 药学院， 浙江 杭州 310053)
[摘要]合成聚乙二醇聚己内酯聚乙烯亚胺(PEGPCLPEI)三嵌段聚合物载体材料，制备包载大黄酸(RH)的PEGPCLPEI纳米粒(PPPRHNPS)，并评价其理化性质和体外生物学特性。通过开环聚合和麦克尔加成反应合成得到PEGPCLPEI聚合物，相对分子质量为95×103，临界胶束浓度为0723 nmol·L-1。包载RH制备得到PPPRHNPS，外观浅黄、乳光明显，透射电镜观察纳米粒分散均匀圆整无团聚，粒径为(1183±36)nm，PDI为(019±008)，Zeta电位为(63±15)mV，包封率为(9364±528)%，载药量为(857±053)%。透析法考察PPPRHNPS体外释药特征，48 h内累积释放率为7592%，释药曲线符合Higuchi模型方程：Q=0121 6t1/2+0069 5(R2=0887 4)，呈缓释特性。选用兔红细胞考察其溶血率，MTT法评价其对HK2细胞的生物安全性，在0～005 mmol·L-1不会引起红细胞溶血和细胞毒性。流式细胞仪考察其摄取效率，PPPRHNPS可被细胞迅速内吞，摄取效率高，30 min内摄取完全，经激光共聚焦显微镜观察，PPPRHNPS能够从溶酶体进入细胞质，具有溶酶体逃逸特性。因此，该研究成功合成PEGPCLPEI聚合物和制备得到PPPRHNPS，粒径分布均匀，包封率及载药量较高，摄取迅速，具有缓释特征、溶酶体逃逸特性、优良的生物安全性，是一种良好研究前景的新型纳米制剂。
, http://www.100md.com
[关键词]大黄酸； 聚乙二醇聚己内酯聚乙烯亚胺； 纳米粒； 体外评价
Preparation and in vitro evaluation of rheinloaded
PEGPCLPEI nanoparticles
CHEN Danfei1， ZHU Yongqin1， ZHANG Yuan1， WANG Linyan2， WEI Yinghui2*
(1. The First Affiliated Hospital of Zhejiang Chinese Medical University， Hangzhou 310006， China；
2College of Pharmaceutical Science， Zhejiang Chinese Medical University， Hangzhou 310053， China)
, 百拇医药
[Abstract]This study was aimed to synthesize the polyethyleneglycolpolycaprolactonepolyethyleneimine (PEGPCLPEI) three block polymer material， prepareRhein (RH)loaded PEGPCLPEI nanoparticles(PPPRHNPS)， and then evaluate their physical and chemical properties and biological characteristics in vitro PEGPCLPEI polymer was obtained by adopting theringopening polymerization and Michael addition reaction， and their physical and chemical properties were analyzed by using NMR and gel permeation chromatography PEGPCLPEI was then used as the carriers to prepare PPPRHNPS by applying spontaneous emulsification solvent diffusion method The results showed that molecular weight of PEGPCLPEI polymer was 95×103， and critical micelle concentration was 0723 mmol·L-1 PPPRHNPS had pale yellow， opalescence faade， round and smooth without aggregation， formed of (1183(36) nm in particle size with PDI of (019±008)， Zeta potential of (63±15) mV， entrapment efficiency of (9364±528)%， and drug loading of (857±053)% The accumulative release percentage of PPPRHNPS was 7592% in 48h， and the release profiles in PBS conformed to the Higuchi equation： Q=0121 6t1/2+0069 5 (R2=0887 4)， presenting slow release characteristics Within the scope of the 0005 mmol·L-1， the nanoparticles had no obvious hemolysis on rabbit red blood cells and toxicity on HK2 cells In the investigation of uptake efficiency by flow cytometry， nanoparticles can be absorbed into cells quickly and internalized within 30 minutes fully， with a high uptake efficiency In confocal laser scanning microscope observation， the nanoparticles can escape from lysosome into cytoplasm Herein， this study synthesized the PEGPCLPEI polymer and prepared PPPRHNPS successfully； the nanoparticles showed uniform particle size， higher encapsulation efficiency and drugloading rate， slow release characteristics， quick uptake and internalization， lysosome escape property and good biocompatibility PPPRHNPS will be a promising pharmaceutical formulation for further development, http://www.100md.com