Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Eur J Pharm Sci. 2018 Jul 1;119:39-48. doi: 10.1016/j.ejps.2018.04.013. Epub 2018 Apr 9.
The purpose of this study was to prepare spray freeze-dried particles of immunoglobulin G (IgG) using various combinations of trehalose and different amino acids (leucine, phenylalanine, arginine, cysteine, and glycine), and investigate the effect of the amino acids on the stability of IgG during the spray freeze-drying (SFD) process and storage. The morphology and structural integrity of the processed particles were evaluated by physical and spectroscopic techniques. SFD-processed IgG without any excipient resulted in the formation of aggregates corresponding to approximately 14% of IgG. In contrast, IgG formulations stabilized using an optimal level of leucine, phenylalanine, or glycine in the presence of trehalose displayed aggregates <2.2%. In particular, phenylalanine combined with trehalose was most effective in stabilizing IgG against shear, freezing, and dehydration stresses during SFD. Arginine and cysteine were destabilizers displaying aggregation and fragmentation of IgG, respectively. Aggregation and fragmentation were evaluated by dynamic light scattering, ultraviolet spectrophotometry, size-exclusion chromatography, and microchip capillary gel electrophoresis. The IgG formulations prepared with leucine, phenylalanine, or glycine in the presence of trehalose showed good stability after storage at 40 °C and 75% relative humidity for 2 months. Thus, a combination of the excipients trehalose and uncharged, nonpolar amino acids appears effective for production of stable SFD IgG formulations.
本研究旨在使用海藻糖和不同氨基酸（亮氨酸、苯丙氨酸、精氨酸、半胱氨酸和甘氨酸）的各种组合来制备免疫球蛋白 G（IgG）的喷雾冷冻干燥颗粒，并研究氨基酸对 IgG 在喷雾冷冻干燥（SFD）过程和储存过程中的稳定性的影响。通过物理和光谱技术评估加工颗粒的形态和结构完整性。没有任何赋形剂的 SFD 处理 IgG 会形成约 14% IgG 的聚集体。相比之下，在海藻糖存在下使用亮氨酸、苯丙氨酸或甘氨酸的最佳水平稳定的 IgG 制剂显示聚集体 <2.2%。特别是，苯丙氨酸与海藻糖结合在 SFD 期间最有效地稳定 IgG 抵抗剪切、冷冻和脱水应激。精氨酸和半胱氨酸分别是导致 IgG 聚集和碎片化的失稳剂。通过动态光散射、紫外分光光度法、尺寸排阻色谱法和微芯片毛细管凝胶电泳评估聚集和碎片化。在海藻糖存在下使用亮氨酸、苯丙氨酸或甘氨酸制备的 IgG 制剂在 40°C 和 75%相对湿度下储存 2 个月后显示出良好的稳定性。因此，赋形剂海藻糖和非带电、非极性氨基酸的组合似乎可有效用于生产稳定的 SFD IgG 制剂。
