title: Freeze-Drying of L-Arginine/Sucrose-Based Protein Formulations, Part 2: Optimization of Formulation Design and Freeze-Drying Process Conditions for an L-Arginine Chloride-Based Protein Formulation System.
abstract: We recently reported that the presence of chloride counter ions in freeze-dried l-arginine/sucrose formulations provided the greatest protein stability, but led to low collapse temperatures and glass transition temperatures of the freeze concentrates. The objectives of this study were to identify l-arginine chloride-based formulations and optimize freeze-drying process conditions to deliver a freeze-dried product with good physical quality attributes (including cake appearance, residual moisture, and reconstitution time). Additional properties were tested such as thermal properties, cake microstructure, and protein physical stability. Excipient concentrations were varied with and without a model protein (bovine serum albumin, BSA). Formulations were frozen with and without annealing or with and without controlled nucleation. Primary drying was conducted at high and low shelf temperature. Cakes with least defects and optimum physical attributes were achieved when protein to excipient ratios were high. Controlled nucleation led to elegant cakes for most systems at a low shelf temperature. Replacing BSA by a monoclonal antibody showed that protein (physical) stability was slightly improved under stress storage temperature (i.e., 40°C) in the presence of a low concentration of l-arginine in a sucrose-based formulation. At higher l-arginine concentrations, cake defects increased. Using optimized formulation design, addition of l-arginine chloride to a sucrose-based formulation provided elegant cakes and benefits for protein stability.
authors.affiliation: GILYOS GmbH, Friedrich-Bergius-Ring 15, Wuerzburg 97076, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Stärtzel
authors.affiliation: GILYOS GmbH, Friedrich-Bergius-Ring 15, Wuerzburg 97076, Germany.
authors.forename: Henning
authors.initials: H
authors.lastname: Gieseler
authors.affiliation: GILYOS GmbH, Friedrich-Bergius-Ring 15, Wuerzburg 97076, Germany.
authors.forename: Margit
authors.initials: M
authors.lastname: Gieseler
authors.affiliation: F. Hoffmann-La Roche Ltd., Pharmaceutical Development and Supplies, Pharma Technical Development Biologics EU, Basel 4070, Switzerland.
authors.forename: Ahmad M
authors.initials: AM
authors.lastname: Abdul-Fattah
authors.affiliation: F. Hoffmann-La Roche Ltd., Pharmaceutical Development and Supplies, Pharma Technical Development Biologics EU, Basel 4070, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Adler
authors.affiliation: F. Hoffmann-La Roche Ltd., Pharmaceutical Development and Supplies, Pharma Technical Development Biologics EU, Basel 4070, Switzerland.
authors.forename: Hanns-Christian
authors.initials: HC
authors.lastname: Mahler
authors.affiliation: F. Hoffmann-La Roche Ltd., Pharmaceutical Development and Supplies, Pharma Technical Development Biologics EU, Basel 4070, Switzerland. Electronic address: pierre.goldbach@roche.com.
authors.forename: Pierre
authors.initials: P
authors.lastname: Goldbach
mesh_terms: D001120:Arginine / Q000737:chemistry*; D002152:Calorimetry, Differential Scanning / Q000379:methods; D002626:Chemistry, Pharmaceutical / Q000379:methods; D002712:Chlorides / Q000737:chemistry*; D003080:Cold Temperature; D003460:Crystallization / Q000379:methods; D004355:Drug Stability; D005079:Excipients / Q000737:chemistry; D005612:Freeze Drying / Q000379:methods; D055550:Protein Stability; D011506:Proteins / Q000737:chemistry*; D013395:Sucrose / Q000737:chemistry*; D044366:Transition Temperature
chemical_list: D002712:Chlorides; D005079:Excipients; D011506:Proteins; D013395:Sucrose; D001120:Arginine
keywords: amorphous; calorimetry (DSC); freeze-drying/lyophilization; glass transition; physical characterization; protein aggregation; protein formulation; solid state stability