D-sn-GPHEEDA的制备及其脂质纳米粒的评价  

Preparation of D-sn-GPHEEDA and Evaluation of its Lipid Nanoparticles

作　　者：刘雅雯 郭思茗 顾乐炎 关丽萍 曹青日[2] LIU Yawen;GUO Siming;GU Leyan;GUAN Liping;CAO Qingri(School of Food Science and Pharmacy of Zhejiang Ocean University,Zhoushan 316022,China;School of Pharmaceutical Sciences,Medical of Soochow University,Suzhou 215123,China)

机构地区：[1]浙江海洋大学食品与药学学院,浙江舟山316022 [2]苏州大学苏州医学院药学院,江苏苏州215123

出　　处：《海南师范大学学报(自然科学版)》2025年第1期22-28,共7页Journal of Hainan Normal University(Natural Science)

基　　金：福建省药物新靶点研究重点实验室开放课题(FJ-YW-2023KF04);浙江省公益技术应用研究计划(2017C33131);江苏省精准诊疗药物创制工程研究中心(发改办高技[2021]1368号)。

摘　　要：优化以二油酰基卵磷脂为原料的1,2-二油酰-sn-甘油-3-磷脂酰羟乙基乙二胺合成工艺,并评价脂质纳米粒的理化特性、siRNA结合力、血清稳定性和细胞毒性。通过HPLC、LC-MS、^(1)H-NMR、^(13)C-NMR表征确证其结构,考察了酶的用量、反应时间、反应物用量、两相反应体系比对1,2-二油酰-sn-甘油-3-磷脂酰羟乙基乙二胺合成的影响。结果表明,最佳反应条件为酶用量150 IU、最佳反应时间72 h、二油酰基卵磷脂与乙二胺的投料比1∶15、乙酸乙酯与乙酸钠缓冲溶液的体积比2∶1时,产率达68.72%。所得脂质纳米粒的粒径为(102.100±2.495)nm,平均多分散指数为0.183±0.023,Zeta电位为+(28.90±2.46)mV,外观呈圆形,稳定性良好;在N/P比为12∶1时可与siRNA完全结合,可显著提高siRNA的血清稳定性;MTT结果表明所合成磷脂及其LNPs对293T细胞活力无显著影响。该研究为其他磷脂酰化合物的合成及纳米递送载体的应用提供参考。In this study,we optimized the synthesis process of D-sn-GPHEEDA from dioleoyllecithin,and evaluated the physicochemical properties,siRNA binding ability,serum stability and cytotoxicity of lipid nanoparticles(LNPs).The structure of D-sn-GPHEEDA was characterized by MS,^(1)H-NMR and^(13)C-NMR,and the effects of enzyme dosage,reaction time,reactant dosage and two reaction systems on the synthesis of D-sn-GPHEEDA were evaluated.The results showed that the optimal reaction conditions were as follows:the enzyme dosage was 150 IU,the reaction time was 72 h,the ratio of dioleyllecithin to ethylenediamine was 1:15,the volume ratio of ethyl acetate to sodium acetate buffer solution was 2:1,and the yield was 68.72%.The prepared LNPs had a spherical appearance and uniform size distribution,with the particle size of(102.100±2.495)nm,PDI of 0.183±0.023,zeta potential of+(28.9±2.46)mV,respectively.The LNPs had good stability and can completely bind to siRNA at N/P ratio of 12∶1 and significantly improve the serum stability of siRNA.Though the MTT assay,the synthesized phospholipid and LNPs showed no significant effects on the viability of 293T cells.This study provides a basis for the synthesis of other phospholipid compounds and the application of LNP-based nanocarriers.

关 键 词：D-sn-GPHEEDA 脂质纳米粒 SIRNA 稳定性 细胞毒性 

分 类 号：R914.5[医药卫生—药物化学] R944.2[医药卫生—药学]