基于PLGA纳米粒的SARS-CoV-2 DNA疫苗的制备及初步评价  

Preparation and preliminary evaluation of SARS-CoV-2 DNA vaccine based on PLGA nanoparticles

作　　者：吴雅琦 李蒙 邢昊楠 陈大全[1] 郑爱萍 WU Ya-qi;LI Meng;XING Hao-nan;CHEN Da-quan;ZHENG Ai-ping(Pharmacy School,Yantai University,Yantai 264005,China;Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)

机构地区：[1]烟台大学药学院,山东烟台264005 [2]军事科学院军事医学研究院毒物药物研究所,北京100850

出　　处：《药学学报》2023年第6期1677-1684,共8页Acta Pharmaceutica Sinica

摘　　要：本研究构建并优化编码新型冠状病毒(severe acute respiratory syndrome coronavirus 2, SARS-CoV-2)刺突(spike protein, S)蛋白基因序列的质粒DNA (plasmid DNA, pDNA) Opt-S,以聚乳酸-羟基乙酸共聚物[poly(lactic-co-glycolic acid), PLGA]纳米粒(nanoparticles, NPs)作为pDNA的递送载体,采用纳米沉淀法制备PLGApDNA NPs,对其体外性质进行初步评价。结果表明所制备的PLGA-pDNA NPs形态规整,边缘清晰,平均粒径为(184.2±2.4) nm,多分散系数(polydisperse index, PDI)为0.093±0.013, zeta电位为(-68.10±0.36) mV,包封率为(98.92±0.22)%,于-20℃储存7个月粒径和PDI变化幅度均较小,稳定性较好,能保护pDNA免受核酸酶降解,并具有一定的缓释效果,细胞毒性低,安全性高,体外转染实验表明SARS-CoV-2 S基因可以进入细胞并表达。以上结果表明PLGA-pDNA NPs非病毒基因载体制备工艺简单、性能良好,有望为SARS-CoV-2疫苗研发提供新思路。We constructed and optimized the plasmid DNA(pDNA)Opt-S encoding the gene of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)spike(S)protein,using poly(lactic-co-glycolic acid)copolymer(PLGA)as a delivery carrier for pDNA.PLGA-pDNA NPs were loaded by nanoprecipitation and its properties in vitro were preliminary evaluated.The results showed that the prepared PLGA-pDNA NPs were regular morphology,clear edges,with an average particle size of(184.2±2.4)nm,polydisperse index(PDI)of 0.093±0.013,zeta potential of(-68.10±0.36)mV,and encapsulation rate of(98.92±0.22)%.The PLGA-pDNA NPs were stable at-20℃for 7 months and could protect pDNA against nuclease degradation.And they also exhibited sustained release of pDNA in vitro.The PLGA-pDNA NPs have low cytotoxicity and high safety.In addition,in vitro transfection experiments showed that the SARS-CoV-2 S gene could enter cells and be expressed.These results indicate that PLGA-pDNA NPs non-viral gene vector have simple preparation process and good performance,which are expected to provide a new idea for the research and development of SARS-CoV-2 vaccine.

关 键 词：新型冠状病毒肺炎 新型冠状病毒DNA疫苗 聚乳酸-羟基乙酸共聚物 纳米沉淀法 非病毒基因载体 

分 类 号：R944[医药卫生—药剂学]