Silicone Oil Particles in Prefilled Syringes With Human Monoclonal Antibody, Representative of Real-World Drug Products, Did Not Increase Immunogenicity in In Vivo and In Vitro Model Systems

Abstract

Silicone oil is a lubricant for prefilled syringes (PFS), a common primary container for biotherapeutics. Silicone oil particles (SiOP) shed from PFS are a concern for patients due to their potential for increased immunogenicity and therefore also of regulatory concern. To address the safety concern in a context of manufacturing and distribution of drug product (DP), SiOP was increased (up to~25,000 particles/mL) in PFS filled with mAb1, a fully human antibody drug, by simulated handling of DP mimicked by drop shock. These samples are characterized in a companion report (Jiao N et al. J Pharm Sci. 2020). The risk of immunogenicity was then assessed using in vitro and in vivo immune model systems. The impact of a common DP excipient, polysorbate 80, on both the formation and biological consequences of SiOP was also tested. SiOP was found associated with (1) minimal cytokine secretion from human peripheral blood mononuclear cells, (2) no response in cell lines that report NF-kB/AP-1 signaling, and (3) no antidrug antibodies or significant cytokine production in transgenic Xeno-het mice, whether or not mAb1 or polysorbate 80 was present. These results suggest that SiOP in mAb1, representative of real-world DP in PFS, poses no increased risk of immunogenicity.

Use of In Vitro Human Skin Models to Assess Potential Immune Activation In Response to Biotherapeutic Attributes and Process-related Impurities

“…This is consistent with our previous findings that the mAb aggregates are mildly and transiently, or negligibly immunogenic in various model systems. [34][35][36][37][38][39][40] It is important to note that the non-stressed mAbs showed a slight increase in metabolic activity and time-dependent increase in cytokine secretion (although the cytokine response was similar to that induced by the formulation buffer). This is consistent with our previous results in cell-based assays using peripheral blood mononuclear cells (PBMC) where non-stressed mAbs have been shown to increase the secretion of cytokines.…”

“…Previous publications have shown that biotherapeutic aggregates, representative of real-world drug products, induce only weak and transient immune responses in in vitro and in vivo model systems. 36,40 In these reports, highly aggregated mAbs (orders of magnitude above levels that are found in drug products) were found to induce a low magnitude of anti-drug antibodies in only a small subset of transgenic mice, with no immunological memory (data not shown), as well as cytokine secretion in cell-based assays. To determine whether the skin model might be sensitive enough to detect aggregated therapeutic mAbs, stirring and pH-induced aggregates were generated and characterized.…”

“…34 We have previously shown that highly aggregated antibodies (orders of magnitude above that found in products), generated by various stress conditions, are able to mildly enhance the immune response in cell-based assays and induce anti-drug antibodies (ADA) in transgenic mice, however the response was weak and transient. [34][35][36][37][38][39][40] Aggregates are a critical quality attribute for biotherapeutics that must be monitored and controlled through drug product development, manufacturing, during lot release, and stability.…”

Frequency and component analysis of contaminants generated in preparation of anticancer agents using closed system drug transfer devices (CSTDs)

“…We speculate that this might explain the high incidence of contamination in cases treated with monoclonal antibodies. On the other hand, immunogenicity in the human body after inadvertent injection of the aggregates formed by the reaction of silicone oil and proteins, including monoclonal antibodies, is not fully understood 32 – 34 .…”

Stress Factors in Primary Packaging, Transportation and Handling of Protein Drug Products and Their Impact on Product Quality

“…10 Furthermore, recent studies did not show enhanced immunogenicity of a monoclonal antibody in the presence of silicone oil droplets. 14,15,65 However, additional work is needed to understand the effect of silicone oil on proteins and immunogenicity.…”