转铁蛋白修饰雷公藤甲素脂质体的制备与体外评价  

Preparation of transferrin modified triptolide liposome and in vitro evaluation

作　　者：杨一帆 梁艺瑶 刘保保 苏雪蓉 乔萍 狄留庆 赵晓莉 YANG Yi-fan;LIANG Yi-yao;LIU Bao-bao;SU Xue-rong;QIAO ping;DI Liu-qing;ZHAO Xiao-li(Jiangsu Engineering Research Center for Efficient Delivery System of TCM,School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学药学院,江苏省中药高效给药系统工程技术研究中心,江苏南京210023

出　　处：《中草药》2022年第3期687-695,共9页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金青年基金项目(81102815)。

摘　　要：目的制备转铁蛋白(transferrin,Tf)修饰的雷公藤甲素(triptolide,TP)脂质体(Tf-TP@Lip),并对其进行质量评价和体外靶向性研究。方法采用薄膜分散法制备Tf-TP@Lip,以包封率和载药量为考察指标,通过单因素考察和星点设计-效应面优化法筛选处方和最佳制备工艺;采用超滤离心法测定药物包封率;透射电子显微镜(TEM)观察脂质体微观形态;激光粒度仪测定脂质体的粒径、多分散指数(polydispersity index,PDI)和Zeta电位;并对其体外释放情况进行考察;采用流式细胞仪和荧光显微镜探究细胞摄取情况。结果优化后Tf-TP@Lip的处方和工艺:脂药比为5.72∶1,脂胆比为8.11∶1,磷脂与DSPE-PEG2000的质量比为6∶1,成膜溶剂为无水乙醇,成膜温度60℃。制得的Tf-TP@Lip平均粒径为(130.33±1.89)nm,PDI为0.19±0.03,Zeta电位为(-23.20±0.64)m V,包封率和载药量分别为(85.33±0.41)%和(9.96±0.21)%,TEM下呈球形,大小均匀、圆整。体外释放研究表明,Tf-TP@Lip相比游离TP具有一定缓释效果。与非靶向雷公藤甲素脂质体(TP@Lip)相比,转铁蛋白的修饰明显增加了SMMC-7721对脂质体的摄取效率。结论制备Tf-TP@Lip稳定、可行,粒径小、外观圆整且具备较高的包封率和载药量,可用于进一步的实验研究。Objective To prepare transferrin(Tf)modified triptolide(TP)liposome(Tf-TP@Lip)and evaluate the quality and in vitro targeting.Methods The Tf-TP@Lip were prepared by thin-film dispersion method.With encapsulation efficiency and drug loading as indicators,single factor experiments and CCD-RSM were used to optimized the formulation and optimum preparation of Tf-TP@Lip;The drug entrapment efficiency was determined by ultrafiltration centrifugation;And the microscopic morphology of liposomes were observed under the transmission electron microscope(TEM);The particle size,polydispersion index(PDI),and Zeta potential were determined by laser particle size analyzer,and investigating the in vitro release of liposomes;flow cytometry and microscope were utilized for exploring the cellular uptake.Results The optimized preparation process and formulation were as follow:lipid/drug ratio was 5.72:1,lipid/cholesterol ratio was 8.11:1,the mass ratio of phospholipid to DSPE-PEG2000 was 6:1,the film-forming solvent was absolute ethanol and the temperature was 60℃.The mean particle size,PDI and Zeta potential of optimized TP@Lip were(130.33±1.89)nm,0.19±0.03,(-23.2±0.64)mV,respectively.The encapsulation efficiency and drug loading were(85.33±0.41)%and(9.96±0.21)%,respectively.The resulting liposomes exhibited spherical shape and were narrow in size distribution.The in vitro release studies showed that Tf-TP@Lip has sustained-release effect compared with free TP.And in contrast of non targeted triptolide liposomes(TP@Lip),Tf modification significantly increased the uptake efficiency of liposomes by SMMC-7721.Conclusion The preparation is stable and feasible,the obtained liposomes have a uniform particle size,round appearance and high encapsulation efficiency and drug loading,which can be used for further research.

关 键 词：雷公藤甲素 转铁蛋白 修饰 脂质体 星点设计-效应面法 质量评价 细胞摄取 体外靶向性 薄膜分散法 超滤离心法 

分 类 号：R283.6[医药卫生—中药学]