Immunoglobulin formulation and method of preparation thereof

Abstract

A stable aqueous pharmaceutical formulation comprising a therapeutically effective amount of an antibody, polysorbate 80, a buffer which inhibits polysorbate oxidation is described along with methods of making the preparation. Also described are formulations with high antibody concentrations which maintain fixed volumes and which may be used on patients of variable weight.

Description

FIELD OF THE INVENTION The invention is directed to stable, concentrated formulations of proteins or antibodies, such as natalizumab, wherein the activity of the antibody is retained and also can be administered in a small volume and can be administered to a subject of variable weight in need thereof. BACKGROUND OF THE INVENTION Antibody and protein formulations are known in the art. However, preparing protein formulations, such as antibody formulations, which are chemically and biologically stable, are fraught with challenges. Preparing formulations which are also not only stable but can maintain a small volume (i.e., allowing for a small volume injection) even with an increased concentration of protein, such as antibody, also is problematic. The need for such formulations exist. For example, concentrated amounts of protein in a fixed volume that is also stable would be especially beneficial to patients of variable weight. Administration of fluids to patients of variable weights ...

Examples

example 1

Selection of Polysorbate 80

The typical method of administering natalizumab is intravenous. Intravenous administration requires the final formulation to be isotonic. A formulation of AN100226 (natalizumab), 5 mg / mL in 50 mM L-histidine, 150 mM NaCl, pH 6.0 was initially chosen (Formulation #1). During a Phase II study, protein precipitation of the antibody was observed during the dilution and introduction of natalizumab into the clinical dosing apparatus. Polysorbate 80 was introduced into the formulation (Formulation #2) to resolve the observed protein precipitation. Preferably, the polysorbate for use with the present invention is low in peroxide, i.e., polysorbate from Sigma, Product number P6479, Lot Number 071K7283.

The two factors that have been shown to accelerate the precipitation of the AN100226 antibody are the presence of trace levels of silicone oil and denaturation at the air-liquid interface. The silicone oil was introduced into the product upon use of standard lubri...

example 2

Selection of The Phosphate Buffer

During release testing of the histidine placebo (containing 0.02% w / v polysorbate 80), new trace impurities were detected. These impurities arose from degradation of polysorbate 80, apparently through an oxidation reaction involving metal ions and histidine. For active natalizumab drug product, these trace impurities have been detected only after storage at elevated temperatures (e.g., 25° C. and 40° C.). Thus, the decision was made to modify the placebo and use phosphate to replace histidine in the buffer. The lot of histidine used in the product placebo was different than that used for the active natalizumab drug product lots AN100226-004 and AN100226-005. The impact of the source of histidine on polysorbate 80 degradation is discussed in greater detail infra.

During testing of the placebo, trace impurities were detected in the histidine / NaCl / 0.02% (w / v) polysorbate 80 placebo (Formulation #2). These impurities were detected by their absorbency ...

example 3

Natalizumab Formulation with Polysorbate 80 and Histidine Combined

The mechanism by which these trace impurities are produced is thought to be through a metal catalyzed oxidation of polysorbate (see Donbrow et al., 1978 J. Pharmaceutical Sciences, 67(12): 28). Donbrow describes that autoxidation during storage occurs for different types of polysorbate (e.g., polysorbate 20). Light, temperatures and metal ions also impact autoxidation. Donbrow et al. (1978). It has been confirmed that both histidine and polysorbate 80 are required for this reaction to proceed at a significant rate. Ajinomoto is the single histidine source used for formulation. However significant differences in the rate of reaction between lots supplied by the Ajinomoto have been observed.

An additional factor that plays a role in accelerating the reaction is the presence of metal. This was demonstrated by running a reaction at 60° C. for five days in glass vessel with 50 mM histidine (Lot No. R016A008) and 2% (w / v...

FIELD OF THE INVENTION The invention is directed to stable, concentrated formulations of proteins or antibodies, such as natalizumab, wherein the activity of the antibody is retained and also can be administered in a small volume and can be administered to a subject of variable weight in need thereof. BACKGROUND OF THE INVENTION Antibody and protein formulations are known in the art. However, preparing protein formulations, such as antibody formulations, which are chemically and biologically stable, are fraught with challenges. Preparing formulations which are also not only stable but can maintain a small volume (i.e., allowing for a small volume injection) even with an increased concentration of protein, such as antibody, also is problematic. The need for such formulations exist. For example, concentrated amounts of protein in a fixed volume that is also stable would be especially beneficial to patients of variable weight. Administration of fluids to patients of variable weights ...

Examples

example 1

Selection of Polysorbate 80

The typical method of administering natalizumab is intravenous. Intravenous administration requires the final formulation to be isotonic. A formulation of AN100226 (natalizumab), 5 mg / mL in 50 mM L-histidine, 150 mM NaCl, pH 6.0 was initially chosen (Formulation #1). During a Phase II study, protein precipitation of the antibody was observed during the dilution and introduction of natalizumab into the clinical dosing apparatus. Polysorbate 80 was introduced into the formulation (Formulation #2) to resolve the observed protein precipitation. Preferably, the polysorbate for use with the present invention is low in peroxide, i.e., polysorbate from Sigma, Product number P6479, Lot Number 071K7283.

The two factors that have been shown to accelerate the precipitation of the AN100226 antibody are the presence of trace levels of silicone oil and denaturation at the air-liquid interface. The silicone oil was introduced into the product upon use of standard lubri...

example 2

Selection of The Phosphate Buffer

During release testing of the histidine placebo (containing 0.02% w / v polysorbate 80), new trace impurities were detected. These impurities arose from degradation of polysorbate 80, apparently through an oxidation reaction involving metal ions and histidine. For active natalizumab drug product, these trace impurities have been detected only after storage at elevated temperatures (e.g., 25° C. and 40° C.). Thus, the decision was made to modify the placebo and use phosphate to replace histidine in the buffer. The lot of histidine used in the product placebo was different than that used for the active natalizumab drug product lots AN100226-004 and AN100226-005. The impact of the source of histidine on polysorbate 80 degradation is discussed in greater detail infra.

During testing of the placebo, trace impurities were detected in the histidine / NaCl / 0.02% (w / v) polysorbate 80 placebo (Formulation #2). These impurities were detected by their absorbency ...

example 3

Natalizumab Formulation with Polysorbate 80 and Histidine Combined

The mechanism by which these trace impurities are produced is thought to be through a metal catalyzed oxidation of polysorbate (see Donbrow et al., 1978 J. Pharmaceutical Sciences, 67(12): 28). Donbrow describes that autoxidation during storage occurs for different types of polysorbate (e.g., polysorbate 20). Light, temperatures and metal ions also impact autoxidation. Donbrow et al. (1978). It has been confirmed that both histidine and polysorbate 80 are required for this reaction to proceed at a significant rate. Ajinomoto is the single histidine source used for formulation. However significant differences in the rate of reaction between lots supplied by the Ajinomoto have been observed.

An additional factor that plays a role in accelerating the reaction is the presence of metal. This was demonstrated by running a reaction at 60° C. for five days in glass vessel with 50 mM histidine (Lot No. R016A008) and 2% (w / v...