Oxidation of acetaminophen to N-acetyl-p-aminobenzoquinone imine by human CYP3A4

Biochem Pharmacol. 1993 Apr 22;45(8):1563-9. doi: 10.1016/0006-2952(93)90295-8.


We have investigated: (a) the formation of N-acetyl-p-aminobenzoquinone imine (NAPQI) from acetaminophen (APAP) by reconstituted human liver CYP3A4, (b) the kinetics of NAPQI formation in microsomes prepared from four human livers varying in CYP1A2, 2E1 and 3A4 content determined by Western blot analysis, (c) the contribution of CYP3A4 to the total formation of NAPQI from 0.1 mM APAP in human liver microsomes using troleandomycin as a specific inhibitor, and (d) the relationship between the contribution of CYP3A4 to NAPQI formation and relative CYP3A4 content. The Km of CYP3A4 for APAP was found to be approximately 0.15 mM, similar to concentrations observed in humans after therapeutic doses of the drug. The kinetics of formation of NAPQI in human liver microsomes were complex; the lower Km was similar to that found for reconstituted CYP3A4. The contribution of CYP3A4 to total NAPQI formation varied from 1 to 20% among livers, and correlated with the relative CYP3A4 content, r2 = 0.88, P < 0.05. Our findings indicate that CYP3A4, the major P450 isoform in human liver and enterocytes, contributes appreciably to the formation of the cytotoxic metabolite NAPQI at therapeutically relevant concentrations of APAP and suggest that APAP may be a previously unrecognized inhibitor of this enzyme.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acetaminophen / metabolism*
  • Acetaminophen / pharmacology
  • Benzoquinones
  • Cytochrome P-450 CYP3A
  • Cytochrome P-450 Enzyme Inhibitors
  • Cytochrome P-450 Enzyme System / metabolism*
  • Ditiocarb
  • Dose-Response Relationship, Drug
  • Humans
  • Imines
  • Intestines / enzymology
  • Isoenzymes / antagonists & inhibitors
  • Kinetics
  • Liver / enzymology*
  • Microsomes, Liver / enzymology
  • Mixed Function Oxygenases / antagonists & inhibitors
  • Mixed Function Oxygenases / metabolism*


  • Benzoquinones
  • Cytochrome P-450 Enzyme Inhibitors
  • Imines
  • Isoenzymes
  • Acetaminophen
  • Cytochrome P-450 Enzyme System
  • Ditiocarb
  • Mixed Function Oxygenases
  • CYP3A protein, human
  • Cytochrome P-450 CYP3A
  • CYP3A4 protein, human
  • N-acetyl-4-benzoquinoneimine