title: Enantioselective binding of disopyramide to alpha1-acid glycoprotein and its variants.
abstract: OBJECTIVE
  alpha1-acid glycoprotein (AAG) has three main genetic variants, F1, S, and A variants. There are few reports on the correlation between AAG variants and binding activity of drug enantiomers. We studied the differences between the binding characteristics of enantiomers of disopyramide (DP), which is a basic drug. The aim of this study was to elucidate the cause of the differences between the binding characteristics of DP enantiomers.
  
  
  METHODS
  The variants in human AAG were separated by hydroxyapatite chromatography. Binding of DP enantiomers to AAG variants was studied by the ultrafiltration method. The characteristics of the binding of DP enantiomers to total variants and each variant were examined by Scatchard analysis within a range of concentrations from 0.5 to 50.0 microg/ml.
  
  
  RESULTS
  The binding capacity of S-DP was significantly higher than that of R-DP in variant 3, although the binding capacities of DP enantiomers were almost the same in variant 2. On the other hand, the binding capacities for both S-DP and R-DP in variant 3 were significantly higher than those in variant 2. Furthermore, there was an almost 2.4-fold difference in the dissociation constant (Kd) between S-DP and R-DP in variant 3, although no significant difference was observed in the number of binding sites (N). In variant 2 no significant differences between DP enantiomers were observed in either the dissociation constant or number of binding sites per molecule of AAG. On the other hand, significant differences between variants 2 and 3 in the dissociation constant for both S-DP and R-DP were observed. The differences in dissociation constant between variants 2 and 3 were 4.0-fold in S-DP and 1.7-fold in R-DP.
  
  
  CONCLUSIONS
  The difference between the binding capacities of S-DP and R-DP is due to differences in the association of DP to variants 3-6, and the role of the variants 1 and 2 in the binding of drugs to AAG is minor.
authors.affiliation: Department of Pharmacy, Hokkaido University Hospital, School of Medicine, Hokkaido University, Kita-14-jo, Nishi-5-chome, Kita-ku, Sapporo 060-8648, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Kishino
authors.forename: S
authors.initials: S
authors.lastname: Itoh
authors.forename: T
authors.initials: T
authors.lastname: Nakagawa
authors.forename: K
authors.initials: K
authors.lastname: Miyazaki
mesh_terms: D000889:Anti-Arrhythmia Agents / Q000737:chemistry*; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D004206:Disopyramide / Q000737:chemistry*; D006801:Humans; D066298:In Vitro Techniques; D009961:Orosomucoid / Q000737:chemistry*; D011485:Protein Binding; D020033:Protein Isoforms; D013237:Stereoisomerism
chemical_list: D000889:Anti-Arrhythmia Agents; D009961:Orosomucoid; D020033:Protein Isoforms; D004206:Disopyramide