Bustami R T, Chan H K, Dehghani F, Foster N R
Faculty of Pharmacy, The University of Sydney, NSW, Australia.
Pharm Res. 2000 Nov;17(11):1360-6. doi: 10.1023/a:1007551006782.
To investigate the feasibility of using the Aerosol Solvent Extraction System (ASES) to generate microparticles of proteins suitable for aerosol delivery from aqueous-based solutions.
The ASES technique using high-pressure carbon dioxide modified with ethanol was utilised for the generation of microparticles of proteins (lysozyme, albumin, insulin and recombinant human deoxyribonuclease (rhDNase)) from aqueous solutions. Particle size, morphology, size distributions and powder aerosol performance were examined. The biochemical integrity of the processed proteins was assessed by testing the level of molecular aggregation using size exclusion chromatography and by bioassay technique for lysozyme.
Proteins were precipitated as spherical particles ranging in size from 100 to 500 nm. The primary nano-sized particles agglomerated to form micron-sized particles during the precipitation process. The median size of the particles was a function of the operating conditions. In-vitro aerosol performance tests showed that the percent fine particle mass (< 5 microm) was approximately 65%, 40% and 20% for lysozyme, albumin and insulin, respectively. Negligible loss in the monomer content or biological activity was observed for lysozyme. Insulin exhibited slight aggregation and 93% of the monomer was retained after processing. Albumin was affected by processing and only 50-75% of the monomer was retained compared with 86% in the original material. However, rhDNase was substantially denatured during processing as shown by the significantly reduced monomer content.
Micron-sized particles of lysozyme, albumin and insulin with satisfactory inhalation performance were successfully generated from aqueous solutions using the modified ASES technique. The biochemical integrity of the processed proteins was a function of the operating conditions and the nature of the individual protein.
研究使用气溶胶溶剂萃取系统（ASES）从水基溶液中生成适合气溶胶递送的蛋白质微粒的可行性。
利用用乙醇改性的高压二氧化碳的ASES技术从水溶液中生成蛋白质（溶菌酶、白蛋白、胰岛素和重组人脱氧核糖核酸酶（rhDNase））微粒。检查了颗粒大小、形态、尺寸分布和粉末气溶胶性能。通过使用尺寸排阻色谱法测试分子聚集水平和通过溶菌酶的生物测定技术评估加工后蛋白质的生化完整性。
蛋白质沉淀为大小在100至500nm范围内的球形颗粒。在沉淀过程中，初级纳米级颗粒聚集形成微米级颗粒。颗粒的中值大小是操作条件的函数。体外气溶胶性能测试表明，溶菌酶、白蛋白和胰岛素的细颗粒质量百分比（<5微米）分别约为65%、40%和20%。溶菌酶的单体含量或生物活性损失可忽略不计。胰岛素表现出轻微聚集，加工后保留了93%的单体。白蛋白受加工影响，与原始材料中的86%相比，仅保留了50-75%的单体。然而，如单体含量显著降低所示，rhDNase在加工过程中基本变性。
使用改进的ASES技术成功地从水溶液中生成了具有令人满意的吸入性能的溶菌酶、白蛋白和胰岛素微米级颗粒。加工后蛋白质的生化完整性是操作条件和单个蛋白质性质的函数。
