人工外泌体共递送siRNA和蛋白的递送系统的设计及体外评价  

Design and in vitro evaluation of delivery systems for co-delivery of siRNA and proteins by mimetic exosomes

作　　者：邓赛 张灵敏 王萍[1] 李仕颖 林潮金 傅小媚 余细勇[1] DENG Sai;ZHANG Ling-min;WANG Ping;LI Shi-ying;LIN Chao-jin;FU Xiao-mei;YU Xi-yong(Guangdong Key Laboratory of Molecular Target&Clinical Pharmacology and the State Key Laboratory of Respiratory Disease Pharmacology Group,Guangzhou Medical University,Guangzhou 511436,China)

机构地区：[1]广州医科大学广东省分子靶标与临床药理学重点实验室呼吸疾病国家重点实验室药理组

出　　处：《药学学报》2020年第1期139-145,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(U1601227,81330007,81700382);广东省科技重点项目(2015B020225006);广东省自然科学基金资助项目(2019A1515012166)

摘　　要：本文制备了人工外泌体,共传递蛋白和核酸,实现多组分药物高效安全共传递。采用阳离子脂质赋形剂二油酰基三甲基铵丙烷(dioleyl trimethylammonium propane, DOTAP)修饰聚乳酸-羟基乙酸共聚物(polylactic acidglycolic acid copolymer, PLGA)基质来设计优化的制剂,双乳化法制备包裹蛋白和核酸的PLGA/DOTAP纳米粒,再用逆相蒸发法制备最外层的膜结构,此膜结构由二棕榈酰磷脂酰胆碱(1, 2-dipalmitoyl-sn-glycero-3-phosphocholine,DPPC)、二油酰磷脂酰胆碱(1,2-dioleoyl-sn-glycero-3-phosphocholine, DOPC)、二硬脂酰磷脂酰胆碱(1,2-distearoylsn-glycero-3-phosphocholine, DSPC)、胆固醇及膜蛋白组成,通过超声分散和挤出的方式形成人工外泌体结构,并分析其物理特性和传递效果性质。结果表明,人工外泌体粒径约为156.13 nm,带负电荷(-18.23±0.57 mV),能高效共传递蛋白和siRNA,且siRNA能高效抑制目标基因Trim 28的表达。这说明人工外泌体模拟了外泌体结构,实现了多组分药物安全高效的共递送。To prepare the mimetic exosomes and co-delivery proteins and nucleic acids, and achieve efficient and safe co-delivery of multi-component drugs, an optimized formulation was designed by modifying a polylactic acid-glycolic acid copolymer(PLGA) matrix with a cationic lipid excipient dioleyl trimethylammonium propane(DOTAP), and a PLGA/DOTAP nanoparticles packaged protein and nucleic acid was prepared by double emulsion method, and the outermost membrane structure prepared by reverse phase evaporation method and consists of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine(DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine(DOPC), 1,2-distearoyl-sn-glycero-3-phosphocholine(DSPC), cholesterol and membrane proteins. The structure of the mimetic exosomes is formed by ultrasonic dispersion and extrusion, and analyzed its characteristics and nature of the transfer effect. The size of mimetic exosomes was about 156.13 nm, with negative charge(-18.23 ± 0.57 mV), and it could efficiently co-transfer protein and siRNA, and siRNA could effectively inhibit the expression of target gene Trim28. The mimetic exosomes simulate the structure of exosomes and achieve safe and efficient co-delivery of multi-component drugs.

关 键 词：人工外泌体 聚乳酸-羟基乙酸共聚物 脂质体 纳米材料 药物递送 

分 类 号：R943[医药卫生—药剂学]