Abstract

AIM
To investigate the stereoselective binding of mexiletine or ketoprofen enantiomers with different recombinant domains of human serum albumin (HSA).
METHODS
Three domains (HSA DOM I, II and III) were expressed in Pichia pastoris GS115 cells. Blue Sepharose 6 Fast Flow was employed to purify the recombinant HSA domains. The binding properties of the standard ligands, digitoxin, phenylbutazone and diazepam, and the chiral drugs to HSA domains were investigated using ultrafiltration. The concentrations of the standard ligands, ketoprofen and mexiletine were analyzed with HPLC.
RESULTS
The recombinant HSA domains were highly purified as shown by SDS-PAGE and Western blotting analyses. The standard HSA ligands digitoxin, phenylbutazone and diazepam selectively binds to DOM I, DOM II and DOM III, respectively. For the chiral drugs, R-ketoprofen showed a higher binding affinity toward DOM III than S-ketoprofen, whereas S-mexiletine bound to DOM II with a greater affinity than R-mexiletine.
CONCLUSION
The results demonstrate that HSA DOM III possesses the chiral recognition ability for the ketoprofen enantiomers, whereas HSA DOM II possesses that for the mexiletine enantiomers.

Links

Publisher Full Text

Authors

Shi D, Jin YX, Tang YH, Hu HH, Xu SY, Yu LS, Jiang HD, Zeng S

Institution

Department of Drug Metabolism and Pharmaceutical Analysis, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.

Source

Acta pharmacologica Sinica 33:5 2012 May pg 710-6

MeSH

Binding Sites
Blotting, Western
Chromatography, High Pressure Liquid
Cloning, Molecular
Electrophoresis, Polyacrylamide Gel
Humans
Isomerism
Ketoprofen
Ligands
Mexiletine
Molecular Structure
Pichia
Protein Binding
Protein Structure, Tertiary
Recombinant Proteins
Serum Albumin
Structure-Activity Relationship
Ultrafiltration

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22555373