title: Addition of Amino Acids to Further Stabilize Lyophilized Sucrose-Based Protein Formulations: I. Screening of 15 Amino Acids in Two Model Proteins.
abstract: In small amounts, the low molecular weight excipients-sorbitol and glycerol-have been shown to stabilize lyophilized sucrose-based protein formulations. The purpose of this study was to explore the use of amino acids as low molecular weight excipients to similarly enhance stability. Model proteins, recombinant human serum albumin and α-chymotrypsin, were formulated with sucrose in combination with one of 15 amino acid additives. Each formulation was lyophilized at 1:1:0.3 (w/w) protein-sucrose-amino acid. Percent total soluble aggregate was measured by size-exclusion chromatography before and after storage at 50 °C for 2 months. Classical thought might suggest that the addition of the amino acids to the sucrose-protein formulations would be destabilizing because of a decrease in the system's glass transition temperature. However, significant improvement in storage stability was observed for almost all formulations at the ratio of amino acid used. Weak correlations were found between the extent of stabilization and both amino acid molar volume and side-chain charge. The addition of amino acids at a modest level generally improves storage stability, often by more than a 50% increase, for lyophilized sucrose-based protein formulations.
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, University of Connecticut, Storrs, Connecticut 06269.
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Forney-Stevens
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, University of Connecticut, Storrs, Connecticut 06269.
authors.forename: Robin H
authors.initials: RH
authors.lastname: Bogner
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, University of Connecticut, Storrs, Connecticut 06269. Electronic address: michael.pikal@uconn.edu.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Pikal
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D004355:Drug Stability; D005612:Freeze Drying; D005990:Glycerol / Q000737:chemistry; D006801:Humans; D011506:Proteins / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry*; D013395:Sucrose / Q000737:chemistry*
chemical_list: D000596:Amino Acids; D011506:Proteins; D011994:Recombinant Proteins; D012709:Serum Albumin; D013395:Sucrose; D005990:Glycerol
keywords: formulation; freeze-drying/lyophilization; protein aggregation; protein formulation; solid-state stability; stabilization