Impact of Different Saccharides on the In-Process Stability of a Protein Drug During Evaporative Drying: From Sessile Droplet Drying to Lab-Scale Spray Drying

Objectives

Solid biopharmaceutical products can circumvent lower temperature storage and transport and increase remote access with lower carbon emissions and energy consumption. Saccharides are known stabilizers in a solid protein produced via lyophilization and spray drying (SD). Thus, it is essential to understand the interactions between saccharides and proteins and the stabilization mechanism.

Methods

A miniaturized single droplet drying (MD) method was developed to understand how different saccharides stabilize proteins during drying. We applied our MD to different aqueous saccharide-protein systems and transferred our findings to SD.

Results

The poly- and oligosaccharides tend to destabilize the protein during drying. The oligosaccharide, Hydroxypropyl β-cyclodextrin (HPβCD) shows high aggregation at a high saccharide-to-protein molar ratio (S/P ratio) during MD, and the finding is supported by nanoDSF results. The polysaccharide, Dextran (DEX) leads to larger particles, whereas HPBCD leads to smaller particles. Furthermore, DEX is not able to stabilize the protein at higher S/P ratios either. In contrast, the disaccharide Trehalose Dihydrate (TD) does not increase or induce protein aggregation during the drying of the formulation. It can preserve the protein’s secondary structure during drying, already at low concentrations.

Conclusion

During the drying of S/P formulations containing the saccharides TD and DEX, the MD approach could anticipate the in-process (in) stability of protein X at laboratory-scale SD. In contrast, for the systems with HPβCD, the results obtained by SD were contradictory to MD. This underlines that depending on the drying operation, careful consideration needs to be applied to the selection of saccharides and their ratios.

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Materials:

The formulation of a highly water-soluble protein (hereon referred to as ‘protein X’) presenting a molecular weight of  ~52 kDa was provided by Takeda (Vienna, Austria) and used in this study. The lyophilized formulation contained the protein of interest in a concentration  ≥80% w/w. Additionally, albumin, polyethylene glycol, polysorbate 80, and salts were also present. The disaccharide Trehalose Dihydrate (hereon referred to as ‘TD’) (Merck KGaA, Germany; Mw = 378.33 g/mol), the cyclic oligosaccharide Hydroxypropyl-β-cyclodextrin, Kleptose® HP ORAL GRADE (hereon referred to as ‘HPβCD’) (Roquette Frères, France; Mw = 1,501 g/mol) and the polysaccharide Dextran 40 EP (hereon referred to as ‘DEX’) (Pharmacosmos A/S, Denmark; Mw = 40,000 g/mol) were selected based on their different molecular weights, structures, and their known potential to stabilize proteins [14, 34,35,36,37].

They were used to prepare aqueous saccharide-protein solutions (further referred to as ‘S/P-formulations’) for miniaturized drying and spray drying (Table II). According to information about TD obtained from literature [34,35,36,37], 321:4 mM was chosen as the upper S/P molar ratio for using this disaccharide. For HPβCD, 61:4 mM was chosen as the upper S/P molar ratio based on available material. For DEX, 6:4 mM was chosen as the upper S/P molar ratio as this represented the maximum solubility with the protein in the formulation. The value of 4 mM of protein used in the formulation instead of 1 mM originates from the fact that 20% w/w was set as a fixed protein concentration for each formulation. Purified water (TKA Wasseraufbereitungssysteme GmbH, Germany) was used as a solvent. A BIO-RAD Gel Filtration Standard (Bio-Rad Laboratories Ges.m.b.H., Austria) was used for the relative quantification of monomer, dimer, and aggregate species by size exclusion chromatography (SEC).

Dieplinger, J., Pinto, J.T., Dekner, M. et al. Impact of Different Saccharides on the In-Process Stability of a Protein Drug During Evaporative Drying: From Sessile Droplet Drying to Lab-Scale Spray Drying. Pharm Res (2023). https://doi.org/10.1007/s11095-023-03498-w

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