Dipeptidyl peptidase IV is a target for covalent adduct formation with the acyl glucuronide metabolite of the anti-inflammatory drug zomepirac

• Published in: Life sciences (1973) 2001-01, Vol.68 (7), p.785-797
• Main Authors: Wang, Min, Gorrell, Mark D., McCaughan, Geoffrey W., Dickinson, Ronald G.
• Format: Article
• Language: English
• Subjects: Acyl glucuronide; Adducts, rats; Animals; Anti-Inflammatory Agents, Non-Steroidal - metabolism; Anti-Inflammatory Agents, Non-Steroidal - pharmacology; Blotting, Western; CHO Cells; Cricetinae; Dipeptidyl Peptidase 4 - metabolism; Dipeptidyl peptidase IV; DNA Adducts - metabolism; Glucuronides - metabolism; Humans; Hydrogen-Ion Concentration; Liver - enzymology; Male; Precipitin Tests; Rats; Rats, Sprague-Dawley; Reactivity; Recombinant Proteins - chemistry; Tolmetin - analogs & derivatives; Tolmetin - metabolism; Tolmetin - pharmacology; Zomepirac
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/S0024-3205(00)00981-4

The nonsteroidal anti-inflammatory drug zomepirac (ZP) is metabolised to a chemically reactive acyl glucuronide conjugate (ZAG) which can form covalent adducts with proteins. In vivo, such adducts could initiate immune or toxic responses. In rats given ZP, the major band detected in liver homogenates by immunoblotting with a polyclonal ZP antiserum was at 110 kDa. This adduct was identified as ZP-modified dipeptidyl peptidase IV (DPP IV) by immunoblotting using the polyclonal ZP antiserum and monoclonal DPP IV antibodies OX-61 and 236.3. In vitro, ZAG, but not ZP itself, covalently modified recombinant human and rat DPP IV. Both monoclonal antibodies recognized DPP IV in livers from ZP- and vehicle-dosed rats. Confirmation that the 110 kDa bands which were immunoreactive with the ZP and DPP IV antibodies represented the same molecule was obtained from a rat liver extract reciprocally immunodepleted of antigens reactive with these two antibodies. Furthermore, immunoprecipitations with OX-61 antibody followed by immunolotting with ZP antiserum, and the reciprocal experiment, showed that both these antibodies recognised the same 110 kDa molecule in extracts of ZP-dosed rat liver. The results verify that DPP IV is one of the protein targets for covalent modification during hepatic transport and biliary excretion of ZAG in rats.