Coumarin substrates for cytochrome P450 2D6 fluorescence assays

A set of nine 4-aminomethyl-7-alkoxycoumarin derivatives was synthesized an d characterized as substrates for O-dealkylation by recombinant cytochrome P450 2D6, a major human enzyme involved in drug metabolism, Enzymatic O-dea lkylation yields 7-hydroxycoumarins, which have useful fluorescence propert ies. The substrates, which differed in substitution at the amino and 7-hydr oxy positions, varied in terms of catalytic efficiency of O-dealkylation an d in their selectivity as substrates for cytochrome P450 2D6 in human liver microsomes. Several of the compounds are useful as cytochrome P450 2D6 sub strates in single-phase, rapid-throughput assays. (C) 2001 Academic Press.