注射用胆固醇基磷酰齐多夫定自组装体冻干粉的规模化制备

Large-scale preparation of 5'-cholesteryl-phosphoryl zidovudine self-assemblies freeze-dried powders for injection

关键词:  药物自组装体  注入法  艾滋病  冻干  灭菌

Key words:  self-assembled drug delivery system (SADDS)  injection method  human immunodeficiency virus (HIV)  lyophilization  sterilization

摘要:

目的 研究注射用胆固醇基磷酰齐多夫定（5’-cholesteryl-phosphoryl zidovudine，CPZ）自组装体冻干粉规模化制备的处方工艺和性质。方法 采用注入法制备CPZ自组装体，将CPZ的乙醇溶液缓慢注入到水溶液中，并经过旋转蒸发、高压均质处理。通过抑制CPZ分子解离，增加自组装体稳定性，优化处方工艺，涉及不同pH的磷酸盐缓冲液、不同pH的醋酸水溶液、甘露醇，并进一步筛选冻干保护剂用量和灭菌方法。用透射电镜观察CPZ自组装体，用纳米激光粒度仪测定其粒径及Zeta电位。结果 CPZ自组装体冻干粉最优处方工艺为在水溶液中加入甘露醇（甘露醇∶CPZ=2∶1），采用0.05%醋酸水溶液作为水相，可连续制备300 mL以上，并保持稳定。制备的自组装体经0.45 μm无菌滤膜过滤灭菌。CPZ冻干粉加水重新分散后，自组装体的粒径为75.17 nm，PDI值为0.48，Zeta电位为-41.2 mV。透射电镜下显示CPZ自组装体为囊泡结构。结论 本研究优化了规模化制备CPZ自组装体冻干粉的处方工艺，为该新型药物传递系统的成功研制打下基础。

Abstract:

OBJECTIVE To study formulation and process method of large-scale preparation of 5'-cholesteryl-phosphoryl zidovudine(CPZ) self-assemblies freeze-dried powders for injection and their characteristics. METHODS CPZ self-assemblies were prepared with an injection method, where the ethanol solution of CPZ was slowly injected into aqueous solutions followed by rotary evaporation and high-pressure homogeneous process. For the sake of inhibition of CPZ dissociation and improvement of self-assemblies' stability, optimization of formulation and preparation methods involved phosphate buffered solutions with different pH, acetate solutions with different pH, and addition of mannitol. Furthermore, the content of lyoprotectants and sterilization method were explored. CPZ self-assemblies were observed with the transmission electron microscopy (TEM). Size and Zeta potential of them were measured with laser scattering size analyzer. RESULTS The optimal formulation and preparation method included a ratio of mannitol/CPZ (2:1), 0.05% acetic acid solutions as the water phase, which made the large-scale (more than 300 mL) production of CPZ self-assemblies possible. The product could keep long-term stability. The self-assemblies were sterilized by filtrating through 0.45 μm membranes. The re-suspended CPZ self-assemblies had the particle size of 75.17 nm, the PDI was 0.48, and the Zeta potential was -41.2 mV. CPZ self-assemblies were spherical vesicles according to the TEM. CONCLUSION CPZ self-assemblies freeze-dried powders are prepared in large scale with the optimized formulation and preparation method. This study provides a basis for successful development of the novel self-assembled drug delivery system.