title: Boronic acid-containing proteasome inhibitors: alert to potential pharmaceutical bioactivation.
abstract: Medicinal chemists try to avoid certain organic functional groups, summarized in an ever-growing list, in order to avoid the potential bioactivation to reactive metabolites. To add to that alert list, we report herein that boronic acid-containing compound structures, such as those found in proteasome inhibitors bortezomib and ixazomib, can become bioactivated to chemically reactive imine amide metabolites. Test compounds, ixazomib and bortezomib, were incubated in vitro using human liver fractions containing cytosol and microsomes (S9) under conventional conditions in the presence of GSH. Metabolites were then analyzed using LC-MS(n) with or without online hydrogen-deuterium exchange (HDX) liquid chromatography coupled with an LTQ-Orbitrap. The exact mass measurements of both the precursor and product ions were acquired through data dependent acquisition and compared with theoretical values of proposed fragment ions. Upon deboronation catalyzed by cytochrome P450 enzymes, both test compounds formed imine amide metabolites that were identified by high resolution exact mass measurements in both normal aqueous and HDX HPLC-MS analysis. GSH conjugates were also identified and were postulated as nucleophilic addition of GSH to the imine amide metabolites. All mass spectrometric and HDX measurements of these GSH conjugates proved that the GSH unit was added to the carbon atom of the imine amide partial structure, hence demonstrating the electrophilic property of these imine amide metabolites. The awareness of the formation of electrophilic imine amide metabolites from boronic acid-containing compounds, where the boron atom is bonded to a carbon atom adjacent to an amide nitrogen, should help in drug candidate design and optimization with regard to avoiding potential bioactivation.
authors.affiliation: Drug Metabolism and Pharmacokinetics, Teva Branded Pharmaceutical Products R&D, Inc. , 145 Brandywine Parkway, West Chester, Pennsylvania 19380, United States.
authors.forename: Austin C
authors.initials: AC
authors.lastname: Li
authors.forename: Erya
authors.initials: E
authors.lastname: Yu
authors.forename: Steven C
authors.initials: SC
authors.lastname: Ring
authors.forename: James P
authors.initials: JP
authors.lastname: Chovan
mesh_terms: D000577:Amides / Q000378:metabolism*; D001711:Biotransformation; D001897:Boronic Acids / Q000493:pharmacokinetics*; D000069286:Bortezomib; D003600:Cytosol / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D008862:Microsomes, Liver / Q000378:metabolism; D061988:Proteasome Inhibitors / Q000493:pharmacokinetics*; D011719:Pyrazines / Q000493:pharmacokinetics*
chemical_list: D000577:Amides; D001897:Boronic Acids; D061988:Proteasome Inhibitors; D011719:Pyrazines; D000069286:Bortezomib; D005978:Glutathione