Determination by NMR of the binding constant for the molecular complex between alprostadil and a-cyclodextrin: Implications for a freeze-dried formulation

Determination by NMR of the binding constant for the molecular complex between alprostadil and a-cyclodextrin: Implications for a freeze-dried formulation
April 12, 2005
Tore Ramstad , Chad E. Hadden, Gary E. Martin, Stanley M. Speaker, Dirk L. Teagarden and Thomas J. Thamann
International Journal of Pharmaceutics
Science Direct
Abstract
A binding constant was determined for the complexation reaction between alprostadil (PGE1) and a-cyclodextrin (a-CD). This constant was used to calculate the fraction PGE1 free upon reconstitution of Caverject dual chamber syringe, indicated for the treatment of erectile dysfunction. The determination was based on the measurement of the chemical shift of the C20 methyl protons of PGE1. The observed chemical shift varies as a linear function of the amount of PGE1 bound. The binding constant was obtained from the binding isotherm, a curve of the observed chemical shift versus free ligand (a-CD) concentration, through the application of non-linear regression analysis. A value K11 = 966 M-1 ? 130 M-1 (2s), measured at 27 ?C, was obtained. This value is in good agreement with those reported in the literature. The percent PGE1 free was subsequently calculated for the reconstituted solution and in the corpora cavernosum after injection. The latter showed PGE1 to be delivered essentially quantitatively to the targeted site.
Keywords: Binding study; Cyclodextrins; NMR; Stabilization; Freeze-drying