Kim Sun J, Kim Chan W
Department of Biotechnology, University of Korea, Seoul, South Korea; Biotech Group, LG Life Sciences Company, Daejeon, South Korea.
Department of Biotechnology, University of Korea, Seoul, South Korea.
J Pharm Sci. 2016 Feb;105(2):613-622. doi: 10.1016/j.xphs.2015.11.046.
The purpose of this study was to develop and characterize a sodium hyaluronate microparticle-based sustained release formulation of recombinant human growth hormone (SR-rhGH) prepared by spray-drying. Compared to freeze-drying, spray-dried SR-rhGH showed not only prolonged release profiles but also better particle property and injectability. The results of size-exclusion high-performance liquid chromatography showed that no aggregate was detected, and dimer was just about 2% and also did not increase with increase of inlet temperature up to 150 °C. Meanwhile, the results of reversed-phase high-performance liquid chromatography revealed that related proteins increased slightly from 4.6% at 100 °C to 6.3% at 150 °C. Thermal mapping test proved that product temperature did not become high to cause protein degradation during spray-drying because thermal energy was used for the evaporation of surface moisture of droplets. The structural characterization by peptide mapping, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and circular dichroism revealed that the primary, secondary, and tertiary structures of rhGH in SR-rhGH were highly comparable to those of reference somatropin materials. The biological characterization by rat weight gain and cell proliferation assays provided that bioactivity of SR-rhGH was equivalent to that of native hGH. These data establish that spray-dried SR-rhGH is highly stable by preserving intact rhGH and hyaluronate microparticle-based formulation by spray-drying can be an alternative delivery system for proteins.
本研究的目的是开发并表征一种通过喷雾干燥制备的基于透明质酸钠微粒的重组人生长激素缓释制剂（SR-rhGH）。与冷冻干燥相比，喷雾干燥的SR-rhGH不仅显示出延长的释放曲线，而且具有更好的颗粒性质和可注射性。尺寸排阻高效液相色谱结果表明未检测到聚集体，二聚体约为2%，并且在入口温度高达150°C时也不会随着温度升高而增加。同时，反相高效液相色谱结果显示相关蛋白从100°C时的4.6%略有增加至150°C时的6.3%。热图谱测试证明在喷雾干燥过程中产品温度不会过高导致蛋白质降解，因为热能用于液滴表面水分的蒸发。通过肽图谱、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和圆二色性进行的结构表征表明，SR-rhGH中rhGH的一级、二级和三级结构与参考生长激素材料的结构高度可比。通过大鼠体重增加和细胞增殖试验进行的生物学表征表明，SR-rhGH的生物活性与天然hGH相当。这些数据表明，喷雾干燥的SR-rhGH通过保留完整的rhGH具有高度稳定性，并且基于透明质酸钠微粒的喷雾干燥制剂可以作为蛋白质的替代递送系统。
