新型戊二酸胆固醇单酯脂质体的制备及其与H22细胞疫苗的耦联  

Preparation of novel glutaryl cholesterol-liposome and its conjugation with H22 cell vaccine

作　　者：程娟[1] 胡向兵[2] 平其能[1] 刘景晶[2] 

机构地区：[1]中国药科大学药剂学教研室,南京210009 [2]中国药科大学生命科学技术学院酶工程实验室,南京210009

出　　处：《中国药科大学学报》2010年第5期441-446,共6页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.30873183)~~

摘　　要：制备能够与细胞疫苗耦联的新型戊二酸胆固醇单酯脂质体,用作疫苗的载体和佐剂,并研究其贮存稳定性及其最佳耦联条件。以戊二酸酐和胆固醇为原料合成戊二酸胆固醇单酯,再以其为原料与磷脂制备戊二酸胆固醇单酯脂质体,采用薄膜分散法制备脂质体,以粒径及多分散指数为指标优化脂质体处方,制得粒径在150-200 nm之间的戊二酸胆固醇单酯脂质体。对脂质体的稳定性和贮存条件进行研究,结果表明冻干脂质体在-20℃可至少保存12个月。以耦联率为指标,对影响戊二酸胆固醇单酯脂质体与H22细胞疫苗耦联的相关因素进行优化,试验结果表明,复溶后加入EDC活化10 min,再加入H22细胞疫苗和S-NHS孵育3 h,得到耦联率为81.4%的H22细胞疫苗-脂质体耦联物。Liposomes are of potential utility in vaccine as the carrier and adjuvant.The aim of this study was to prepare novel glutaryl cholesterol-liposomes which could be conjugated with the protein vaccine,and to study the storage and conjugation condition of liposomes.Glutaryl-cholesterol was synthesized using glutarate and cholesterol and identified by thin-layer chromatography（TLC）,mass spectrometry（MS） and elemental analysis.Based on the synthesized glutaryl-chloesterol and phospholipids,glutaryl cholesterol-liposome was prepared using thin-film dispersion,and the formulation of the liposomes was optimized regarding the particle size and polydispersity index.The resultant glutaryl cholesterol-liposomes under the optimal condition have the particle size of 150-200 nm.In addition,the stability of the optimized glutaryl cholesterol-liposomes was observed under the selected temperature.It was shown that lyophilized glutaryl cholesterol-liposomes could be stable at-20 °C up to 12 months.Furthermore,the conditions for the liposomal conjugation with H22 cell vaccine were evaluated to maximize the conjugation efficiency.After aqueous reconstitution,the liposomes were activated for 10min by adding N-ethyl-N′-（3-dimethylaminopropyl） carbodiimide（EDC）,and then incubated for 3 h with N-hydroxy S-succinimide（S-NHS） and the vaccine.The H22 cell vaccine-conjugated liposome was obtained with the conjugation efficiency of 81.4%.Therefore,novel glutaryl cholesterol-liposomes with the relatively better stability and significant capability of conjugation with the protein vaccine were successfully prepared.

关 键 词：戊二酸胆固醇 戊二酸胆固醇单酯脂质体 H22细胞疫苗 耦联 

分 类 号：R944[医药卫生—药剂学] R979.5[医药卫生—药学]