Evidence that 4-allyl-o-quinones spontaneously rearrange to their more electrophilic quinone methides: potential bioactivation mechanism for the hepatocarcinogen safrole

Chem Res Toxicol. May-Jun 1994;7(3):443-50. doi: 10.1021/tx00039a024.


Several naturally occurring aromatic ethers, of which safrole [1-allyl-3,4-(methylenedioxy)-benzene] is one example, are hepatocarcinogens. One bioactivation pathway previously proposed for safrole involves hydroxylation of the benzyl carbon, conjugation with sulfate, and then alkylation of DNA with displacement of the sulfate group [Miller, J.A., and Miller, E.C. (1983) Br. J. Cancer 48, 1-15]. The fact that safrole is O-dealkylated to the corresponding catechol (hydroxychavicol, 1-allyl-3,4-dihydroxybenzene) indicates that quinoid formation is also possible and may contribute to the genotoxic and/or cytotoxic activity of this compound. In the present investigation we selectively oxidized hydroxychavicol to the corresponding o-quinone (HC-quinone, 4-allyl-3,5-cyclohexadiene-1,2-dione) or p-quinone methide (HC-QM, 2-hydroxy-4-allylidene-2,5-cyclohexadien-1-one) and trapped these reactive electrophiles with glutathione (GSH). The GSH adducts were fully characterized by UV, NMR, and mass spectrometry. Microsomal incubations with safrole or hydroxychavicol in the presence of glutathione produced only o-quinone glutathione conjugates. However, if the trapping agent (GSH) was added after an initial incubation of 10 min, both o-quinone and p-quinone methide GSH conjugates were observed. The first-order rate constant of isomerization was estimated from the decrease in HC-quinone GSH adducts to be 1.9 x 10(-3) s-1 (t1/2 = 9 min). Kinetic studies showed that while HC-QM reacts rapidly with water, the model o-quinone (4-tert-butyl-3,5-cyclohexadiene-1,2-dione), which cannot isomerize to a quinone methide, was remarkably resistant to hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Publication types

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

MeSH terms

  • Animals
  • Biotransformation
  • Carcinogens / chemistry*
  • Cytochrome P-450 Enzyme System / metabolism
  • Glutathione / metabolism
  • In Vitro Techniques
  • Isomerism
  • Kinetics
  • Liver Neoplasms, Experimental / chemically induced
  • Male
  • Microsomes, Liver / enzymology
  • Microsomes, Liver / metabolism
  • Monophenol Monooxygenase / metabolism
  • Oxidation-Reduction
  • Oxides / chemistry
  • Quinones / chemistry*
  • Rats
  • Rats, Sprague-Dawley
  • Safrole / chemistry*
  • Safrole / toxicity
  • Silver Compounds / chemistry
  • Spectrophotometry, Ultraviolet


  • Carcinogens
  • Oxides
  • Quinones
  • Silver Compounds
  • disilver oxide
  • Cytochrome P-450 Enzyme System
  • Monophenol Monooxygenase
  • Glutathione
  • Safrole