Stabilization of IgG1 in spray-dried powders for inhalation

Abstract

The protein stabilizing capabilities of spray-dried IgG1/mannitol formulations were evaluated. The storage stability was tested at different residual moisture levels prepared by vacuum-drying or equilibration prior to storage. Vacuum-drying at 32 °C/0.1 mbar for 24 h reduced the moisture level below 1%, constituting an optimal basis for improved storage stability. The crystalline IgG1/mannitol powders with a weight ratio of 20/80 up to 40/60 failed to prevent the antibody aggregation as assessed by size exclusion chromatography during storage. Ratios of 60/40 up to 80/20 IgG1/mannitol provided superior stability of the antibody and the powders could be produced with high yields. The lower the residual moisture, the better was the stabilizing capability. An amount of 20% mannitol provided the best stabilization. Storage stability of 60/40, 70/30, and 80/20 IgG1/mannitol formulations over one year was adequate at 2–8 °C and 25 °C. Closed storage (sealed in vials) at 40 °C/75% RH and open storage at 25 °C/60% RH revealed that the stability still required optimization. The lower the protein content, the better was the powder flowability. The aerodynamic properties of powders spray-dried with 10% solids content were inadequate, as the particle size ranged between 5.1 and 7.2 μm and the fine particle fraction accounted for only 4–11%. Reduction of the solids content to 2.5% did improve the aerodynamic properties as the mass mean aerodynamic diameter was reduced to 3.6 μm and the fine particle fraction was increased to about 14%. The reduction of the solids content did not influence the storage stability significantly. Also spray-drying at higher temperatures had no significant impact on the storage stability, despite a higher tendency to form amorphous systems. In order to improve the storage stability and to maintain the good flowability of 70/30 IgG1/mannitol powder or to keep the storage stability but to improve the flowability of the 80/20 IgG1/mannitol powder, mannitol was partially substituted by a second excipient such as trehalose, sucrose, glycine, lactose, lactosucrose, or dextran 1. Differences in the stabilizing capability were noticeable upon closed storage at 40 °C/75% RH and open powder storage. Protein stabilization was improved by the addition of glycine but trehalose and sucrose were most effective in preventing aggregation, which can be primarily attributed to the water replacement properties of the sugars. The addition of another excipient, isoleucine had positive effects on both flowability and protein stability.