Abstract
PurposeTo measure aggregate and particle formation in tumor necrosis factor-alpha (TNF-α) inhibitors etanercept, adalimumab and certolizumab pegol product samples after exposure to freezing temperature conditions similar to storage conditions previously observed in patients’ homes.MethodsTNF-α inhibitors in their original primary and secondary packaging were exposed to 32 freeze-thaw cycles (−10°C for 120min/5°C for 60 min) or continuous low storage temperature (−20°C for 96 h) before thawing at 2–8°C. Non-stressed products were used as controls. The products were analyzed by high pressure size exclusion chromatography (HP-SEC), dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), micro-flow imaging (MFI) and second derivative ultraviolet (UV) spectroscopy.ResultsTen out of twenty-one stressed product samples (47.6%) showed increased particle numbers in the submicron and micron size range when compared to controls. For each product, DLS, MFI and NTA detected an increase in particle level in at least one stressed syringe (both continuous freezing and freeze-thaw), whereas HP-SEC and UV spectroscopy showed no differences between stressed and non-stressed products.ConclusionTNF-α inhibitors are relatively resistant to freezing temperatures similar to storage conditions previously observed in patients’ homes. However, almost half of the stressed product samples showed formation of particles in the submicron and micron size range.
Highlights
The introduction of drugs containing tumor necrosis factoralpha (TNF-α) inhibitors has revolutionized treatments for many inflammatory diseases such as rheumatoid arthritis and inflammatory bowel disease [1]
Two product samples showed an increase in Z-average and polydispersity index (PdI) (product sample E3: Z-average 17.48 (SD 0.01)/PdI 0.27 (SD 0.01); product sample B3: Z-average 24.00 (SD 0.03)/PdI 0.27 (SD 0.03)) after multiple freeze-thaw stress conditions
Additional peaks in size distribution were detected after both stress conditions; product samples E2, E3, B3 exposed to multiple freeze-thaw stress conditions and product samples E4, Page 5 of 11 42
Summary
The introduction of drugs containing tumor necrosis factoralpha (TNF-α) inhibitors has revolutionized treatments for many inflammatory diseases such as rheumatoid arthritis and inflammatory bowel disease [1]. A previous study showed that most patients do not store TNF-α inhibitors within this recommended temperature range; only 7% of patients were able to store TNF-α inhibitors continuously between 2 and 8°C [7]. Almost 25% of patients stored their TNF-α inhibitors below 0°C for 2 h or longer; 5.9% of patients stored their TNF-α inhibitors below 0°C for at least 24 h, with the lowest temperature measured around −20°C. Almost 14% of the patients exposed their TNF-α inhibitors to at least three re-current freeze-thaw cycles with a median duration of almost 4 days. Six patients (2.4%) even exposed their drugs to at least 32 recurrent freeze-thaw cycles [7]. The most common consequence of exposing proteins to freezing temperature conditions is the formation of aggregates [8,9] which may lead to the development of antidrug antibodies and decreased drug effectiveness, as well as an increased probability of side effects [10,11]
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