Activity-stability considerations of trypsinogen during spray drying: effects of sucrose.
The preparation and processing of protein pharmaceuticals into powders may impose significant stresses that could perturb and ultimately denature them. In many cases their stabilization through added excipients is necessary to yield native and active proteins. In this study, the effect of spray drying on the structure and activity of a model protein (trypsinogen) was investigated. In the absence of excipients, spray drying resulted in small losses of its enzymatic activity. Protein conformational rearrangements in the solid state (observed via FTIR) and irreversible aggregation (upon reconstitution) constituted the major degradation pathways. The irreversible unfolding in the solid state was also confirmed by solution calorimetric studies that indicated a decreased thermal stability of the spray-dried protein after reconstitution. The presence of sucrose, a thermal and dehydration stress stabilizer, induced a concentration-dependent protective effect. Protein protection was afforded even at low carbohydrate concentrations, while at specific mass ratios (sucrose-to-protein = 1:1) complete activity preservation was achieved. However, at the high end of sucrose concentrations, a small destabilization was evident, indicating that excluded volume effects may be undesirable during preparation of protein microparticles via spray drying. The profile of both the protein conformational changes and thermal stability in the solid state closely followed that of the incurred activity losses, indicating that protein stabilization during dehydration is crucial during processing of these polypeptides.
Tzannis
S T
ST
Biopharmaceutical R&D, ALZA Corporation, 950 Page Mill Road, Palo Alto, California 94303, USA. stelios.tzannis@inhale.com
Prestrelski
S J
SJ
D002626:Chemistry, Pharmaceutical; D002850:Chromatography, Gel; D004355:Drug Stability; D006358:Hot Temperature; D010316:Particle Size; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D013395:Sucrose / Q000737:chemistry*; D014362:Trypsinogen / Q000737:chemistry*
D013395:Sucrose; D014362:Trypsinogen