Vol. 28, Issue 7, 715-717, July 2000

ACCELERATED COMMUNICATION
Selective and Potent Inhibition of Human CYP2C19 Activity by a Conformationally Targeted Antipeptide Antibody

Timothy Schulz-Utermoehl, Richard J. Mountfield, Kjeld Madsen, Peer N. Jørgensen, and Kristian T. Hansen

Departments of Drug Metabolism (T.S.-U., R.J.M., K.T.H.) and Medicinal Chemistry (K.M.), Novo Nordisk A/S, Novo Nordisk Park, Maaloev, Denmark; and Department of Immunochemistry (P.N.J.), Novo Nordisk A/S, Gentofte, Denmark

A conformationally targeted anti-peptide antibody was produced by immunizing a rabbit with a cyclized peptide corresponding to a loop region of human CYP2C19 (residues 250-261). In an enzyme-linked immunosorbent assay, the antibody bound strongly to recombinant CYP2C19 and poorly to recombinant CYP2C8, CYP2C9, and CYP2C18. In immunoblotting studies, the antibody bound strongly to recombinant CYP2C19 and weakly to recombinant CYP2C8. No binding to recombinant CYP1A2, CYP2C9, CYP2C18, CYP2D6, CYP2E1, and CYP3A4 was detected. In immunoinhibition experiments, the anti-peptide antibody targeted against CYP2C19 potently inhibited (S)-mephenytoin 4'-hydroxylase activity of human hepatic microsomal fraction (>90%). It had no appreciable effect on ethoxyresorufin O-deethylase (CYP1A2), tolbutamide methyl-hydroxylase (CYP2C9), dextromethorphan O-demethylase (CYP2D6), 4-nitrophenol hydroxylase (CYP2E1), or testosterone 6-hydroxylase (CYP3A4) activity of human hepatic microsomal fraction. However, large amounts of purified IgG fractions were able to inhibit up to 35% of paclitaxel 6-hydroxylase (CYP2C8) activity. In conclusion, we have demonstrated that an anti-peptide antibody targeted against residues 250 to 261 of human CYP2C19 selectively and potently inhibited CYP2C19 activity of human hepatic microsomal fraction.