Effect of viscosity on the deamidation rate of a model Asn-hexapeptide.
The effect of viscosity on the deamidation rate of a model Asn-containing hexapeptide (l-Val-l-Tyr-Pro-l-Asn-Gly-l-Ala) was assessed in aqueous solution and in solids containing varying amounts of poly(vinyl pyrrolidone) (PVP) and water. Stability studies were conducted at 0.1 mg/mL peptide and 0-50% PVP (w/w) in aqueous solution, and at 5% (w/w) peptide and different relative humidities (31.6, 53.1, 74.4 and 96%) in the solid state. The parent peptide and its deamidation products were analysed by reverse-phase high-performance liquid chromatography. Deamidation rates decreased with increasing solvent viscosity in a manner described by a semi-empirical mathematical model developed to describe this relationship. The results suggest that the motion of the Asn side-chain along the reaction coordinate is a function of the macroscopic solvent viscosity. However, the apparent energy barrier for the diffusive movement of the side-chain appears to be less than the energy barrier for that associated with macroscopic viscosity. The dependence of the deamidation rate on viscosity in both viscous solution and hydrated solids further demonstrates the importance of mobility in peptide deamidation.
Li
R
R
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66047, USA.
Hageman
M J
MJ
Topp
E M
EM
D002152:Calorimetry, Differential Scanning; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D008956:Models, Chemical; D008970:Molecular Weight; D009842:Oligopeptides / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry*; D011205:Povidone; D012996:Solutions; D014783:Viscosity
D009842:Oligopeptides; D010446:Peptide Fragments; D012996:Solutions; C089005:valyl-tyrosyl-prolyl-asparaginyl-glycyl-alanine; D011205:Povidone