Dixit Nitin, Salamat-Miller Nazila, Salinas Paul A, Taylor Katherine D, Basu Sujit K
Drug and Combination Product Development, Shire, Lexington, Massachusetts 02421.
Drug and Combination Product Development, Shire, Lexington, Massachusetts 02421.
J Pharm Sci. 2016 May;105(5):1657-1666. doi: 10.1016/j.xphs.2016.02.029. Epub 2016 Mar 28.
This study investigated the root cause behind an observed free fatty acid particle formation and resulting Polysorbate 20 (PS20) loss for a sulfatase drug product upon long-term storage at 5 ± 3°C. Reversed- phase chromatography with mass spectrometric analysis as well as charged aerosol detection was used to characterize the peaks associated with the intact and degraded PS20. Additionally, a proteomics study was undertaken to identify the residual host cell proteins in the sulfatase drug substance. PS20 stability studies were conducted in the presence of sulfatase, a sulfatase inhibitor, putative phospholipase B-like 2, and mock drug substance produced using a null cell line vector under experimental conditions optimized for PS20 degradation. This study provides the first published evidence where the residual host cell protein present in the drug substance was identified and experimentally shown to catalyze the breakdown of PS20 in a protein formulation over time, resulting in free fatty acid particles and PS20 loss. This study demonstrates the importance of early detection of potential impurities in the protein drug substance that may contribute to polysorbate degradation to make a judicious selection of the surfactant and its optimized concentration for the final drug product.
本研究调查了一种硫酸酯酶药物产品在5±3°C长期储存后观察到的游离脂肪酸颗粒形成以及由此导致的聚山梨酯20（PS20）损失背后的根本原因。采用反相色谱结合质谱分析以及带电气溶胶检测来表征与完整和降解的PS20相关的峰。此外，还进行了一项蛋白质组学研究，以鉴定硫酸酯酶药物原料中的残留宿主细胞蛋白。在硫酸酯酶、一种硫酸酯酶抑制剂、假定的磷脂酶B样2以及使用空细胞系载体在针对PS20降解优化的实验条件下生产的模拟药物原料存在的情况下，进行了PS20稳定性研究。本研究提供了首个已发表的证据，即鉴定出药物原料中存在的残留宿主细胞蛋白，并通过实验证明其随着时间的推移催化蛋白质制剂中PS20的分解，导致游离脂肪酸颗粒和PS20损失。本研究证明了早期检测可能导致聚山梨酯降解的蛋白质药物原料中潜在杂质的重要性，以便为最终药物产品明智地选择表面活性剂及其优化浓度。
