Coumarin substrates for cytochrome P450 2D6 fluorescence assays

Anal Biochem. 2001 May 15;292(2):280-6. doi: 10.1006/abio.2001.5098.

Abstract

A set of nine 4-aminomethyl-7-alkoxycoumarin derivatives was synthesized and characterized as substrates for O-dealkylation by recombinant cytochrome P450 2D6, a major human enzyme involved in drug metabolism. Enzymatic O-dealkylation yields 7-hydroxycoumarins, which have useful fluorescence properties. The substrates, which differed in substitution at the amino and 7-hydroxy positions, varied in terms of catalytic efficiency of O-dealkylation and in their selectivity as substrates for cytochrome P450 2D6 in human liver microsomes. Several of the compounds are useful as cytochrome P450 2D6 substrates in single-phase, rapid-throughput assays.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Catalysis / drug effects
  • Coumarins / chemical synthesis
  • Coumarins / chemistry*
  • Coumarins / metabolism*
  • Cytochrome P-450 CYP2D6 / chemistry
  • Cytochrome P-450 CYP2D6 / isolation & purification
  • Cytochrome P-450 CYP2D6 / metabolism*
  • Cytochrome P-450 CYP2D6 Inhibitors
  • Enzyme Inhibitors / pharmacology
  • Fluorescent Dyes / chemical synthesis
  • Fluorescent Dyes / chemistry*
  • Fluorescent Dyes / metabolism*
  • Humans
  • Kinetics
  • Microsomes, Liver / drug effects
  • Microsomes, Liver / enzymology
  • Microsomes, Liver / metabolism
  • Molecular Sequence Data
  • Quinidine / pharmacology
  • Recombinant Proteins / antagonists & inhibitors
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Substrate Specificity

Substances

  • Coumarins
  • Cytochrome P-450 CYP2D6 Inhibitors
  • Enzyme Inhibitors
  • Fluorescent Dyes
  • Recombinant Proteins
  • coumarin
  • Cytochrome P-450 CYP2D6
  • Quinidine