Mittag Judith J, Trutschel Marie-Luise, Kruschwitz Helen, Mäder Karsten, Buske Julia, Garidel Patrick
Boehringer Ingelheim Pharma GmbH & Co. KG, Innovation Unit, PDB-TIP, Birkendorfer Straße 65, 88397 Biberach an der Riss, Germany.
Martin-Luther-University Halle-Wittenberg, Institute of Pharmacy, Faculty of Biosciences, Wolfgang-Langenbeck-Strasse 4, 06120 Halle (Saale), Germany.
Int J Pharm X. 2022 Jun 23;4:100123. doi: 10.1016/j.ijpx.2022.100123. eCollection 2022 Dec.
Polysorbates are an important class of nonionic surfactants that are widely used to stabilize biopharmaceuticals. The degradation of polysorbate 20 and 80 and the related particle formation in biologics are heavily discussed in the pharmaceutical community. Although a lot of experimental effort was spent in the detailed study of potential degradation pathways, the underlying mechanisms are only sparsely understood. Besides enzymatic hydrolysis, another proposed mechanism is associated with radical-induced (auto)oxidation of polysorbates. To characterize the types and the origin of the involved radicals and their propagation in bulk material as well as in diluted polysorbate 80 solutions, we applied electron paramagnetic resonance (EPR) spectroscopy using a spin trapping approach. The prerequisite for a meaningful experiment using spin traps is an understanding of the trapping rate, which is an interplay of (i) the presence of the spin trap at the scene of action, (ii) the specific reactivity of the selected spin trap with a certain radical as well as (iii) the stability of the formed spin adducts (a slow decay rate). We discuss whether and to which extent these criteria are fulfilled regarding the identification of different radical classes that might be involved in polysorbate oxidative degradation processes. The ratio of different radicals for different scenarios was determined for various polysorbate 80 quality grades in bulk material and in aqueous solution, showing differences in the ratio of present radicals. Possible correlations between the radical content and product parameters such as the quality grade, the manufacturing date, the manufacturer, the initial peroxide content according to the certificate of analysis of polysorbate 80 are discussed.
聚山梨酯是一类重要的非离子表面活性剂，广泛用于稳定生物制药。聚山梨酯20和80的降解以及生物制品中相关颗粒的形成在制药界受到广泛讨论。尽管在详细研究潜在降解途径方面投入了大量实验工作，但对其潜在机制的了解仍然很少。除了酶促水解外，另一种提出的机制与聚山梨酯的自由基诱导（自）氧化有关。为了表征所涉及自由基的类型、来源及其在散装材料以及稀释的聚山梨酯80溶液中的传播，我们采用自旋捕获方法应用电子顺磁共振（EPR）光谱。使用自旋捕获进行有意义实验的前提是了解捕获速率，捕获速率是以下因素的相互作用：（i）自旋捕获剂在作用现场的存在，（ii）所选自旋捕获剂与特定自由基的特定反应性，以及（iii）形成的自旋加合物的稳定性（缓慢衰减速率）。我们讨论了在鉴定可能参与聚山梨酯氧化降解过程的不同自由基类别方面，这些标准是否以及在多大程度上得到满足。测定了散装材料和水溶液中不同质量等级的聚山梨酯80在不同情况下不同自由基的比例，结果表明存在的自由基比例存在差异。还讨论了自由基含量与产品参数之间的可能相关性，如质量等级、生产日期、制造商、根据聚山梨酯80分析证书的初始过氧化物含量。
