Lin Zhang1,2, Sandeep Yadav3, Y John Wang3, Olivier Mozziconacci1,4, Christian Schӧneich5. 1. Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, 66047, USA. 2. Present address: Eli Lilly and Company, Lilly Corporate Center, Indianapolis, Indiana, 46285, USA. 3. Late Stage Pharmaceutical Development, Genentech Inc., South San Francisco, California, 94080, USA. 4. Merck Research Laboratories, Rahway, New Jersey, 07065, USA. 5. Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, 66047, USA. schoneic@ku.edu.
Abstract
PURPOSE: L-Histidine (L-His) and polysorbate 20 (PS20) are two excipients frequently included in parenteral products to stabilize biotherapeutics. The objective of the current work was to investigate the impact of L-His on PS20 stability in aqueous solutions when subjected to forced oxidation and accelerated stability testing. METHODS: The stability of PS20 in L-His buffer was compared with that in acetate buffer. Forced oxidation of PS20 in these two buffer systems was initiated by a free radical generator, 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH), while accelerated stability tests were carried out at 40°C. Ultra-performance liquid chromatography mass spectrometry was utilized to monitor intact PS20 and to analyze degradation products. RESULTS: Our results demonstrate a dual effect of L-His on PS20 stability. During exposure to AAPH, L-His protects PS20 from oxidation. Stable isotope labeling of L-His with 13C was employed for mechanistic investigations. The protection of L-His was abrogated when acetate was added to L-His buffer, implying that the anti-oxidative activity of L-His may be compromised by specific counter ions. The replacement of L-His by various His derivatives led to significant changes in the protection of PS20 against AAPH-induced degradation. In contrast to forced degradation, the addition of L-His promoted oxidative PS20 degradation during accelerated storage at 40°C in solution, generating mainly short chain POE-laurates. CONCLUSION: L-His exhibits a dual effect on the stability profile of PS20, protecting against AAPH-induced oxidation but promoting oxidative degradation during accelerated stability testing.
PURPOSE:L-Histidine (L-His) and polysorbate 20 (PS20) are two excipients frequently included in parenteral products to stabilize biotherapeutics. The objective of the current work was to investigate the impact of L-His on PS20 stability in aqueous solutions when subjected to forced oxidation and accelerated stability testing. METHODS: The stability of PS20 in L-His buffer was compared with that in acetate buffer. Forced oxidation of PS20 in these two buffer systems was initiated by a free radical generator, 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH), while accelerated stability tests were carried out at 40°C. Ultra-performance liquid chromatography mass spectrometry was utilized to monitor intact PS20 and to analyze degradation products. RESULTS: Our results demonstrate a dual effect of L-His on PS20 stability. During exposure to AAPH, L-His protects PS20 from oxidation. Stable isotope labeling of L-His with 13C was employed for mechanistic investigations. The protection of L-His was abrogated when acetate was added to L-His buffer, implying that the anti-oxidative activity of L-His may be compromised by specific counter ions. The replacement of L-His by various His derivatives led to significant changes in the protection of PS20 against AAPH-induced degradation. In contrast to forced degradation, the addition of L-His promoted oxidative PS20 degradation during accelerated storage at 40°C in solution, generating mainly short chain POE-laurates. CONCLUSION:L-His exhibits a dual effect on the stability profile of PS20, protecting against AAPH-induced oxidation but promoting oxidative degradation during accelerated stability testing.
Authors: Lin Zhang; Sandeep Yadav; Barthélemy Demeule; Y John Wang; Olivier Mozziconacci; Christian Schӧneich Journal: Pharm Res Date: 2016-10-13 Impact factor: 4.200
Authors: Ravuri S K Kishore; Sylvia Kiese; Stefan Fischer; Astrid Pappenberger; Ulla Grauschopf; Hanns-Christian Mahler Journal: Pharm Res Date: 2011-03-03 Impact factor: 4.200
Authors: Vanessa V Agon; William A Bubb; Adam Wright; Clare L Hawkins; Michael J Davies Journal: Free Radic Biol Med Date: 2005-12-12 Impact factor: 7.376