Acetaminophen binds to mouse hepatic and renal DNA at human therapeutic doses.
Alkylation of DNA by acetaminophen metabolites has been reported previously, but has received little attention, and the biological impact of this alkylation is essentially unknown. In the present study, apparent covalent binding of acetaminophen metabolites to DNA in male ICR mice was observed at levels of 2.0 +/- 0.4 to 18.5 +/- 5.5 pmol of acetaminophen/mg of DNA in liver and 0.6 +/- 0.1 to 26.9 +/- 2.6 pmol of acetaminophen/mg of DNA in kidney with doses ranging from 10 to 400 mg/kg. Investigations of the reaction of [3H]-N-acetyl-p-benzoquinone imine (NAPQI) or [ring-14C]NAPQI with DNA in vitro yielded low levels of DNA alkylation. Greater apparent binding of [3H]NAPQI to DNA occurred in reactions containing nuclear proteins, such as by using chromatin or whole nuclei. The binding of NAPQI to purified DNA also was enhanced by the presence of 0.1 mM cysteine, but not by 1.0 mM cysteine. Increased binding of NAPQI to DNA in the presence of cysteine or nuclear protein is in contrast to the effects of alternate sulfhydryls on the binding of NAPQI to proteins, which implies that the mechanisms responsible for binding to DNA may be different than the mechanisms that mediate alkylation of protein. The alkylation of DNA by [ring-14C]NAPQI was enhanced markedly at buffer pH < 4.0, suggesting participation of a protonated form of NAPQI in binding to DNA under these conditions. Acetaminophen binding to DNA also was assessed in metabolic activation systems, including microsomes with cumene hydroperoxide or NADPH, and with horseradish peroxidase (HRP) and H2O2. Measurable binding was obtained in all systems, but HRP and H2O2 produced binding levels 200-fold greater than was observed with the microsomal systems. The 32P-postlabeling of DNA from acetaminophen-treated mice, and of DNA reacted with acetaminophen, HRP, and H2O2, produced unique spots that were not identical. The present data further support the hypothesis that acetaminophen metabolites bind covalently to DNA and demonstrate that this apparent binding is observed in experimental animals in vivo at doses that mimic therapeutic doses in humans.
Rogers
L K
LK
Department of Pediatrics, Baylor College of Medicine, Houston, Texas 77030, USA. lrogers@neomailbcem.pedi.bcem.tmc.edu
Moorthy
B
B
Smith
C V
CV
D000082:Acetaminophen / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR
D000082:Acetaminophen; D004247:DNA