Some properties of an inhibitory cytochrome P-450 metabolic-intermediate complex existing in a spin-state equilibrium.
Incubation, in the presence of NADPH/O2, of the type II compound revenast with a partially solubilized, phenobarbital-induced rat liver microsomal cytochrome P-450 system results in the formation of a difference spectrum exhibiting a Soret band at 407 nm and a trough at 423 nm. Experiments with N2 and metyrapone suggest this spectral perturbation to originate from binding to the hemoprotein of a metabolic intermediate derived from the amine substrate. The percentage of pigment complexed can be assessed to be about 7%, with half-maximal complexation occurring at a revenast concentration of 125 microM. Adduct formation is inhibitory to the N-hydroxylation of 4-chloroaniline and N-demethylation of N,N-dimethylaniline; kinetic analysis suggests a competitive interaction at the catalytic site of the reactive revenast derivative with the tertiary arylamine. Inhibition of the two monooxygenation reactions is reversible, indicating weak heme bonding of the active intermediate. This behaviour differs from that of most other product adducts so far examined. The aberrant functional properties of the metabolic adduct appear to reflect complex molecular organization of this compound, as is also evidenced by the unique position of the Soret region absorption maximum at 407 nm; this spectral data hints at the presence of a mixture of high- and low-spin forms. Indeed, chemical reduction or oxidation of the product complex reveals the existence of a low-spin component hidden in the metabolic spectrum. Model studies suggest that the product adduct possibly arises from N-oxidation of the revenast molecule.
Walther Straub-Institut für Pharmakologie und Toxikologie der Universität, Munich, Federal Republic of Germany.
P
P
Hlavica
U
U
Künzel
G
G
Achtert
D000814:Aniline Compounds / Q000378:metabolism; D000818:Animals; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D006633:Histamine Antagonists / Q000378:metabolism*; D008297:Male; D010084:Oxidation-Reduction; D011720:Pyrazoles / Q000378:metabolism*; D047069:Pyrazolones; D011725:Pyridines / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013053:Spectrophotometry
D000814:Aniline Compounds; D065607:Cytochrome P-450 Enzyme Inhibitors; D006633:Histamine Antagonists; D011720:Pyrazoles; D047069:Pyrazolones; D011725:Pyridines; C054026:revenast; C015157:N,N-dimethylaniline; D003577:Cytochrome P-450 Enzyme System; C004658:4-chloroaniline