抗动脉粥样硬化核酸疫苗纳米冻干剂的研究

Study on Lyophilized Nanoparticles of Antiatherosclerotic DNA Vaccine

目的:碱裂解法提取编码胆固醇酯转运蛋白的质粒(简称pCETP)作为核酸疫苗,并制成适合鼻腔递送的载核酸疫苗壳聚糖纳米冻干剂。方法:碱裂解法大量提取抗动脉粥样硬化核酸疫苗,以壳聚糖(简称CS)为非病毒载体,复凝聚法制备pCETP/CS纳米粒,正交设计筛选最佳pCETP/CS纳米冻干剂处方。结果:提取了纯度可靠的pCETP,制得的pCETP/CS冻干剂具有较高的包封率(94.6%)和载药量(27.3%),重分散后纳米粒粒径为(375±18)nm,pCETP浓度可达4000μg.mL-1,较冻干前提高40倍。结论:pCETP/CS冻干剂具有良好的稳定性和重分散性,为适合鼻内递送的载基因微粒制剂,具有较好的研究前景。

Objective： The plasmid encoding cholesterol ester transfer protein （pCETP） extracted by alkaline lysis approach, was used to prepare pCETP/Chitosan（CS） lyophilized nanopartieles for nasal mucosa administration. Methods： Alkaline lysis method was used for extracting pCETP as antiatheroselerosis DNA vaccine. The pCETP loaded chitosan nanoparticles were prepared by the complex eoacervation method. Results： The obtained purity of pCETP was reliable. By the orthogonal design, the optimized freeze-drying formulation led to high encapsulating efficiency（94.6%） and drug-loading content（27.3%）. The size of the redispersed nanoparticles was （375±18）nm, and the concentration of pCETP was as high as 4000μg·ml^-1. Furthermore, the volume of the administration was reduced as much as 40 fold, compared with the volume before freeze-drying. Conclusion： The pCETP/CS freeze-dried complex with good stability and redispersibility is a potential immunization formulation for nasal mucosa administration and worthy for further exploitation.