磷酸奥司他韦-乳铁蛋白吸入剂的制备及HPAEpiC细胞毒性与抑菌作用研究

采用喷雾干燥法制备了磷酸奥司他韦-乳铁蛋白干粉吸入剂。以m(L-亮氨酸)∶m(甘露醇)、溶液体积、进料速率、气体流量为影响因素，以收率、卡尔指数(Carr’s index)、空气动力学直径(D_a)、体外肺部细微粒子沉积率(FPF)为主要指标，设计了4因素3水平的正交试验，对制剂配方及喷雾干燥工艺参数进行优化，得到优化后的制备条件为：m(L-亮氨酸)∶m(甘露醇)=3∶0,溶液体积40 mL,进料速率约6.4 mL/min,气体流量约473 L/h。采用CCK-8法测试在优化条件下制备的干粉吸入剂对正常人肺泡上皮细胞(HPAEpiC)的毒性，并采用梯度稀释法和菌落计数法测试了干粉吸入剂对金黄色葡萄球菌(Staphylococcus aureus)的抑制作用，结果表明：当干粉吸入剂的质量浓度不高于600μg/mL(所含磷酸奥司他韦的质量浓度约为353μg/mL)时，HPAEpiC细胞的存活率大于90%,表明制备的干粉吸入剂在此浓度范围内对HPAEpiC细胞几乎没有损伤作用；在质量浓度为1 mg/mL时干粉吸入剂对金黄色葡萄球菌具有一定的抑制作用，抑菌率为22.50%。

Preparation of oseltamivir phosphate-lactoferrin inhalation and its cytotoxicity to HPAEpiC cells and antibacterial effect

Oseltamivir phosphate-lactoferrin dry powder inhalation has been prepared by a spray drying method. With the m (L-leucine)∶m (mannitol) ratio, solution volume, feed rate and gas flow as the influencing factors, and the yield, Carr's index, aerodynamic diameter (D_a) and in vitro lung fine particle fraction (FPF) as the main indexes, four factors and three levels of orthogonal tests were designed to optimize the formulation and spray drying process parameters. The optimized preparation conditions were as follows: m (L-leucine)∶m (mannitol)=3∶0, a solution volume of 40 mL, a feed rate of about 6.4 mL/min, and a gas flow of about 473 L/h. The CCK-8 method was used to test the toxicity of the dry powder inhalation prepared under the optimized conditions on normal human alveolar epithelial cells (HPAEpiC). The gradient dilution method and colony counting method were used to test the inhibitory effect of the dry powder inhalation on Staphylococcus aureus. The results showed that when the mass concentration of dry powder inhalation was not higher than 600 μg/mL (the mass concentration of oseltamivir phosphate was about 353 μg/mL), the survival rate of HPAEpiC cells was more than 90%, indicating that the prepared dry powder inhalation had little toxicity to HPAEpiC cells in this concentration range. At a mass concentration of 1 mg/mL, the dry powder inhalation had a certain effect on Staphylococcus aureus, with an inhibitory rate of 22.50%.