Kizuki Shinji, Wang Zekun, Torisu Tetsuo, Yamauchi Satoru, Uchiyama Susumu
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan; Formulation Research Lab., Taiho Pharmaceutical Co. Ltd., 224-2, Ebisuno, Hiraishi, Kawauchi-cho, Tokushima, 771-0194, Japan.
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
J Pharm Sci. 2023 Feb;112(2):492-505. doi: 10.1016/j.xphs.2022.09.022. Epub 2022 Sep 24.
An increase in protein aggregates during transportation should be suppressed in therapeutic protein products because the aggregates have a potential risk of immunogenicity. In this study, three protein solutions in vials were exposed to tri-axial vibration with various combinations of frequency and acceleration using a transportation test system to investigate the relationship between low g-force stresses and protein aggregate generation. The number concentration of micron aggregates detected by flow imaging analysis increased markedly when the acceleration and frequency of agitation were within a specific range, in other words, above a threshold. This threshold was common among the three protein solutions. The suppression of micron aggregate formation by adding a surfactant suggested that agitation above the threshold increased micron aggregates mainly via interface-mediated routes. Notably, agitation, including agitation below the threshold, accelerated spontaneous oligomerization (nanometer aggregate generation) of proteins in bulk solution even in the presence of the surfactant. Studies of stability against mechanical stresses (e.g., a random vibration test to simulate actual shipment, with a time-compressed setting by increasing acceleration) need to be performed and discussed with careful consideration of the threshold for generating micron aggregates.
治疗性蛋白质产品在运输过程中蛋白质聚集体的增加应受到抑制，因为这些聚集体具有潜在的免疫原性风险。在本研究中，使用运输测试系统，将小瓶中的三种蛋白质溶液暴露于频率和加速度的各种组合的三轴振动下，以研究低重力应力与蛋白质聚集体生成之间的关系。当搅拌的加速度和频率在特定范围内，即高于阈值时，通过流动成像分析检测到的微米级聚集体的数量浓度显著增加。这个阈值在三种蛋白质溶液中是共同的。添加表面活性剂对微米级聚集体形成的抑制表明，高于阈值的搅拌主要通过界面介导的途径增加微米级聚集体。值得注意的是，即使存在表面活性剂，包括低于阈值的搅拌也会加速本体溶液中蛋白质的自发寡聚化（纳米级聚集体生成）。需要进行针对机械应力的稳定性研究（例如，通过增加加速度进行时间压缩设置的模拟实际运输的随机振动测试），并在仔细考虑产生微米级聚集体的阈值的情况下进行讨论。
