title: Investigating the role of 2-phenylpropenal in felbamate-induced idiosyncratic drug reactions.
abstract: Felbamate (2-phenyl-1,3-propanediol dicarbamate, FBM) can cause aplastic anemia and hepatotoxicity. The mechanism of FBM-induced toxicities is unknown; however, it has been proposed that 2-phenylpropenal, a reactive metabolite of FBM, is responsible. The pathway leading to this metabolite involves hydrolysis of FBM to 2-phenyl-1,3-propandiol monocarbamate (MCF), oxidation to 3-carbamoyl-2-phenylpropionaldehyde (CBMA), and spontaneous loss of carbon dioxide and ammonia. We made a polyclonal antibody against 2-phenylpropenal bound to protein and confirmed its specificity using ELISA. We attempted to develop an animal model of FBM-induced aplastic anemia and/or hepatotoxicity, and we also used the antibody to try to detect covalent binding of 2-phenylpropenal using immunoblotting. However, none of the animals developed evidence of bone marrow or liver toxicity, and we were unable to detect covalent binding, possibly because significantly less 2-phenylpropenal is formed in rodents than in humans. As this type of idiosyncratic drug reaction is believed to be immune-mediated, we also studied the potential of FBM and its metabolites to stimulate an immune response using the reporter antigen popliteal lymph node assay in female Balb/c mice. We found that neither FBM nor MCF induced an immune response in popliteal lymph nodes (PLNs). However, CBMA treatment appeared immunogenic, causing footpad inflammation, hardening, scab formation, and an increase in thickness. The PLN cell count in CBMA-treated mice increased 8-fold as compared to control, FBM-, or MCF-treated mice. Immunohistochemical analysis of the CBMA-exposed PLNs revealed germinal center formation, indicating B cell proliferation, later confirmed by flow cytometry. Most of the cells expressing the activation surface marker CD54 were B cells. We also found that CBMA treatment caused an increase in the production of IgM and IgG1 antibodies as well as IL-4 and IFN-gamma cytokines. Our findings indicate that 2-phenylpropenal is a very potent immunogen, supporting its possible involvement in the FBM-induced hepatotoxicity and aplastic anemia.
authors.affiliation: Faculty of Pharmacy, University of Toronto, Toronto, Ontario M5S 2S2, Canada.
authors.forename: M
authors.initials: M
authors.lastname: Popović
authors.forename: S
authors.initials: S
authors.lastname: Nierkens
authors.forename: R
authors.initials: R
authors.lastname: Pieters
authors.forename: J
authors.initials: J
authors.lastname: Uetrecht
mesh_terms: D000447:Aldehydes / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D000741:Anemia, Aplastic / Q000139:chemically induced* / Q000150:complications; D000818:Animals; D000927:Anticonvulsants / Q000633:toxicity*; D056486:Chemical and Drug Induced Liver Injury; D000078328:Felbamate; D005260:Female; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D007150:Immunohistochemistry; D007371:Interferon-gamma / Q000276:immunology; D015847:Interleukin-4 / Q000096:biosynthesis / Q000276:immunology; D008099:Liver / Q000187:drug effects*; D008107:Liver Diseases / Q000150:complications; D008198:Lymph Nodes / Q000166:cytology / Q000276:immunology; D051379:Mice; D023421:Models, Animal; D048448:Phenylcarbamates; D011409:Propylene Glycols / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*
chemical_list: D000447:Aldehydes; D000927:Anticonvulsants; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D048448:Phenylcarbamates; D011409:Propylene Glycols; D015847:Interleukin-4; D007371:Interferon-gamma; C013026:hydratropic aldehyde; D000078328:Felbamate