Deamidation of model β-turn cyclic peptides in the solid state

To investigate the importance of secondary structure on peptide deamidation in the solid state, two cyclic β-turn peptides and their linear analogs were used as models of Asn residues in structured and unstructured domains, and incorporated into poly(vinyl pyrrolidone) (PVP)-based lyophilized solids. The secondary structure of the model peptides was determined in solution and the solid state using a combination of nuclear magnetic resonance (NMR) spectroscopy, circular dichroism (CD), and Fourier transform infrared (FTIR) spectroscopy. The model β-turn cyclic peptides were found to be type II β-turns while the linear analogs were determined to be predominantly unstructured. Quantitatively, the cyclic peptides consisted of approximately 80% β-turn while the linear analogs contained only 30%-35% β-turn. To characterize the solid environment, Tg, and moisture content of the solid-state formulations were determined. Accelerated stability studies were conducted in the solid state at 37°C using formulations lyophilized from solutions at pH 8.8 (0.1 M borate buffer). The effect of matrix mobility on solid-state deamidation was investigated by altering the moisture content through variation of relative humidity or the addition of a plasticizer. Cyclic peptides degraded 1.2-8 times slower than the linear analogs under all of the conditions studied. The observed rate constants, however, for all of the peptides decreased dramatically (four orders of magnitude) in the glassy solids. This suggests the greater importance of matrix mobility in solid-state degradation. Molecular dynamics (MD) simulations were also performed to explore the low energy, preferred state of the peptides, and determine the structure around the β-turn. © 2005 Wiley-Liss, Inc. and the American Pharmacists Association.