Assessment of the Immunogenicity of Mechanically Induced Interferon Aggregates in a Transgenic Mouse Model

作者：Human Paul; Ilsley Helen; Roberson Cynthia; Grovender Eric; Van Antwerp Bill; Fogt Eric; Zilla Peter^*

来源：Journal of Pharmaceutical Sciences, 2015, 104(2): 722-730.

DOI：10.1002/jps.24292

• drug delivery systems
• HPLC (high performance
• pressure liquid chromatography)
• capillary electrophoresis
• immunology
• mechanical activation
• protein aggregation
• protein delivery
• stability
• transgenics

Pump delivery of human interferon alpha-2B (IFN2b) has the potential for inducing immunogenic drug aggregates. We therefore evaluated the immunogenicity of mechanically induced IFN2b aggregates to assess this risk. Transgenic human-IFN2b (TG) and wild-type (WT) FVB/N mice (n = 8 and n = 9/group, respectively) were administered mechanically agitated drug [45 Hz for 6 h (LLA) or 24 h (HLA)], chemically modified drug [low pH (pH 4.0) or metal oxidized (OXD)] or unstressed drug (native). Mice received IFN2b (50 g; 100 g/mL; s.c.) formulations on days 0, 7, 14, and 21. Drug-binding and neutralizing antibody titers were determined after 28 d. Aggregate concentrations were highest in OXD and HLA formulations but OXD had more dimers/trimers. Geometric mean titers were 1:131, 1:728, 1:1573, 1:871, and 1:10,240 for WT mice (n = 9) and 1:207, 1:587, 1:1810, 1:571, and 1:2,153 for TG mice (n = 8) for native, LLA, HLA, pH4, and OXD groups, respectively. Mechanical agitation of IFN2b induced equivalent titers of immunoglobulin to that of metal oxidation, both capable of binding to or neutralizing the drug in WT and TG mice. Thus, by limiting metal contamination and by inclusion of a stabilizing agent to mitigate drug aggregation, the risk of anti-drug immunoglobulin may be reduced in a pump delivery scenario.

• 出版日期2015-2