Legumain酶和线粒体双级靶向去氢骆驼蓬碱脂质体的制备及其体外特性评价  

Preparation of Legumain enzyme and mitochondrial double-stage targeted harmine liposome and evaluation of its in vitro characterization

作　　者：伊帕尔古丽·阿皮孜 贺宏吉 王昭志 李喆喆 卡迪热娅·艾克拉木 白静雅 王梅[1] Ipargul·Hafiz;HE Hongji;WANG Zhaozhi;LI Zhezhe;Kadirya·Akram;BAI Jingya;WANG Mei(College of Pharmacy,Xinjiang Medical University,Urumqi 830017,China)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830017

出　　处：《中国药房》2022年第13期1565-1572,共8页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81760637);新疆维吾尔自治区天山英才计划第三期资助项目;新疆天然活性组分和释药技术重点实验室项目(No.XJDX1713);新疆医科大学大学生创新训练计划项目(No.CX2021048)。

摘　　要：目的制备Legumain酶和线粒体双级靶向去氢骆驼蓬碱(HM)脂质体(KA@HM-LPS),并对其制剂学特性、体外抗肿瘤作用及生物相容性进行初步评价。方法首先,筛选KA@HM-LPS的制备方法和均质化方法,并对制备的脂质体进行表征。其次,分别测定KA@HM-LPS的血清稳定性、体外释放率、溶血百分数,以及空白脂质体作用下的细胞存活率。最后,分别测定KA@HM-LPS作用下的细胞存活率、线粒体靶向性和对肝癌细胞迁移和侵袭的抑制作用。结果选择薄膜分散法制备KA@HM-LPS,其包封率为(90.50±0.62)%;选择挤出法作为KA@HM-LPS的均质化方法。所制备的KA@HM-LPS粒径、多分散系数、Zeta电位分别为(211.40±11.67)nm、0.316±0.014和(-14.20±0.49)mV。在37℃、10%胎牛血清中,12 h后KA@HM-LPS的粒径基本稳定;KA@HM-LPS在20%血浆中的体外释放曲线符合Weibull分布,具有缓释效果;当HM质量浓度为160μg/mL时,KA@HM-LPS的溶血百分数为(4.23±0.19)%,远小于游离HM,具有安全性。当空白脂质体的质量浓度达400μg/mL时,LO2细胞的存活率为(94.40±6.12)%,载体生物相容性较好。体外细胞实验结果显示,KA@HM-LPS对Legumain酶过表达的肝癌细胞LGMN+-SK-Hep-1的抑制作用显著高于对正常肝癌细胞SK-Hep-1的抑制作用;且与SK-Hep-1细胞相比,LGMN+-SK-Hep-1细胞对载体的摄取效率更高;KA@HM-LPS可以更明显地抑制LGMN+-SK-Hep-1细胞的迁移和侵袭。结论成功制备KA@HM-LPS;该脂质体可以有效抑制Legumain酶过表达的肝癌细胞的迁移和侵袭,并提高HM的血液相容性。OBJECTIVE To prepare Legumain enzyme and mitochondrial double-stage targeted harmine(HM)liposome(KA@HM-LPS)and preliminary evaluate its pharmaceutical properties,in vitro antitumor effect and biocompatibility.METHODS Firstly,the preparation and homogenization methods of KA@HM-LPS was screened,and prepared liposomes were characterized.Secondly,the serum stability,in vitro release rate,hemolysis percentage of KA@HM-LPS and cell survival rate under KA@BLPS were determines respectively.Finally,the cell surivival rate,mitochondrial targeting and inhibitory effects on cell migration and invasion of KA@HM-LPS were determined.RESULTS KA@HM-LPS was prepared by the thin-film dispersion method,with encapsulation efficiency of(90.50±0.62)%.The extrusion moulding method was selected as homogenization method of [e-mail] particle size,polydispersity index,and Zeta potential of KA@HM-LPS were(211.40±11.67)nm,0.316±0.014 and(-14.20±0.49)mV,respectively.In 37℃,10%FBS,the particle size of KA@HM-LPS kept stable after 12 h.In vitro release curve of KA@HM-LPS in 20%plasma conformed to Weibull distribution and had the property of sustained release.When HM concentration was 160μg/mL,the hemolysis percentage of KA@HM-LPS was(4.23±0.19)%,which was much lower than that of free HM,with safety.When the mass concentration of KA@BLPS reaches 400μg/mL,the survival rate of LO2 cells was(94.40±6.12)%,and the biocompatibility was good.Cell test results in vitro showed that,inhibitory effect of KA@HM-LPS on liver cancer cells with overexpression of Legumain enzyme(LGMN+-SK-Hep-1)was significantly higher than that of normal liver cancer cells SK-Hep-1;compared with SK-Hep-1,LGMN+-SK-Hep-1 cells had a higher uptake efficiency of the liposome;KA@HM-LPS could significantly inhibit the migration and invasion of LGMN+-SK-Hep-1 cells.CONCLUSIONS KA@HM-LPS is prepared successfully,which can effectively inhibit the migration and invasion of liver cancer cells with Legumain enzyme overexpression,and improve the blood compatibility of HM.

关 键 词：去氢骆驼蓬碱 线粒体 Legumain酶 脂质体 

分 类 号：R944[医药卫生—药剂学]