The microsomal dealkylation of N,N-dialkylbenzamides

Biochem Pharmacol. 1994 Jan 20;47(2):275-80. doi: 10.1016/0006-2952(94)90017-5.

Abstract

The in vitro metabolism of N,N-dialkylamides by phenobarbital-induced rat liver microsomes yields an N-alkylamide and the corresponding aldehyde. Although, N-hydroxymethyl-N-alkylamide intermediates can be detected from N-methyl-N-alkylamides, no N-hydroxyalkyl-N-alkylamide intermediates are detected from the N,N-dialkylamide substrates. Vmax values were independent of amide structure, whereas Vmax/Km values were dependent on the lipophilicity of the N,N-dialkylbenazamide studied. These results suggest that diffusion of substrate into the membrane-bound enzyme active site limits the rate of microsomal oxidation of the amides. Metabolism of N-alkyl-N-methylamides reveals identical values of Vmax for demethylation and dealkylation. Values of Vmax/Km for demethylation depend upon the lipophilicity of the N-alkyl group, whereas Vmax/Km values for dealkylation appear to be dependent upon the steric bulk of the alkyl group, particularly around the alpha-carbon. Moreover, Vmax/Km values for demethylation are larger than for dealkylation, implying the reactions are under kinetic control. Comparison of the kinetic data with theoretical AM1 semi-empirical molecular orbital calculations suggests a mechanism involving formation of a carbon-centred radical. Use of an N-cyclopropylmethylbenzamide substrate to trap such a radical failed, presumably because oxygen rebound is faster than radical rearrangement. An N-cyclopropylamide substrate did not undergo metabolism of the cyclopropyl ring, consistent with carbon-centred radical, but not nitrogen radical cation, formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkylation
  • Amidohydrolases / metabolism
  • Animals
  • Benzamides / chemical synthesis
  • Benzamides / chemistry
  • Benzamides / metabolism*
  • Benzoates / analysis
  • Benzoic Acid
  • Cytochrome P-450 Enzyme System / metabolism
  • Kinetics
  • Methylation
  • Microsomes, Liver / enzymology*
  • Oxidoreductases, N-Demethylating / metabolism
  • Phenobarbital
  • Rats
  • Substrate Specificity

Substances

  • Benzamides
  • Benzoates
  • Benzoic Acid
  • Cytochrome P-450 Enzyme System
  • Oxidoreductases, N-Demethylating
  • Amidohydrolases
  • Phenobarbital