title: Activation of the carcinogen N-hydroxy-N-(2-fluorenyl)benzamide via chemical and enzymatic oxidations. Comparison to oxidations of the structural analogue N-hydroxy-N-(2-fluorenyl)acetamide.
abstract: Chemical or enzymatic oxidations of the carcinogen N-hydroxy-N-(2- fluorenyl)benzamide (N-OH-2-FBA) were investigated under the conditions facilitating one-electron oxidation or oxidative cleavage of N-hydroxy-N-(2-fluorenyl)acetamide (N-OH-2-FAA). HPLC methods were developed for separation and quantitation of the above hydroxamic acids and their respective oxidation products. To identify the products of oxidation of N-OH-2-FBA, N-(benzoyloxy)-2-FBA (N-BzO-2-FBA) was synthesized and shown to undergo ortho rearrangement to 1- and 3-BzO-2-FBA. Oxidation of N-OH-2-FBA (4.88 mM) with alkaline K3Fe(CN)6 in benzene was complete and yielded equimolar amounts of 2-nitrosofluorene (2-NOF) and the ester (chiefly N-BzO-2-FBA), indicative of one-electron oxidation to nitroxyl free radical which undergoes bimolecular dismutation. However, one-electron oxidation of N-OH-2-FBA (30 or 10 microM) by horseradish peroxidase/H2O2 at pH 7 or myeloperoxidase/H2O2 at pH 6.5 yielded only approximately 10% as much product as N-OH-2-FAA (30 microM). The addition of 0.1 mM Br- +/- 0.1 M Cl- at pH 4 to 6.5 increased 2-NOF formation in MPO/H2O2-catalyzed oxidations. Simulations of these oxidations with HOCl/Cl- or HOBr/Br- showed that the latter was more efficient, converting N-OH-2-FAA almost completely and less than or equal to 62% of N-OH-2-FBA to 2-NOF. The amounts of the ester (N- and o-BzO-2-FBA), which by itself did not contribute to 2-NOF formation or significant substrate regeneration, indicated that approximately 10% of 2-NOF originated from one-electron oxidation.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Laboratory for Cancer Research, Veterans Affairs Medical Center, Minneapolis, Minnesota 55417.
authors.forename: D
authors.initials: D
authors.lastname: Malejka-Giganti
authors.forename: C L
authors.initials: CL
authors.lastname: Ritter
authors.forename: R W
authors.initials: RW
authors.lastname: Decker
mesh_terms: D001711:Biotransformation; D002273:Carcinogens / Q000737:chemistry* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006735:Horseradish Peroxidase / Q000378:metabolism; D006881:Hydroxyacetylaminofluorene / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000378:metabolism
chemical_list: D002273:Carcinogens; C077090:N-hydroxy-N-(2-fluorenyl)benzamide; D006881:Hydroxyacetylaminofluorene; D006735:Horseradish Peroxidase; D009195:Peroxidase