Study on the regioselectivity and mechanism of the aromatic hydroxylation of monofluoroanilines.
The in vitro and in vivo metabolism of monofluoroanilines was investigated. Special attention was focused on the regioselectivity of the aromatic hydroxylation by cytochromes P-450 and the mechanism by which this reaction might proceed. The results clearly demonstrate that the in vitro and in vivo regioselectivity of the aromatic hydroxylation by cytochromes P-450 is dependent on the fluoro-substituent pattern of the aromatic aniline-ring. Results from experiments with liver microsomes from differently pretreated rats demonstrate that the observed regioselectivity for the aromatic hydroxylation is not predominantly determined by the active site of the cytochromes P-450. To investigate the underlying reason for the observed regioselectivity, semi-empirical molecular orbital calculations were performed. Outcomes of these calculations show that neither the frontier orbital densities of the LUMO/LUMO + 1 (lowest unoccupied molecular orbital) of the monofluoroanilines nor the spin-densities in their NH. radicals can explain the observed regioselectivities. The frontier orbital densities of the HOMO/HOMO - 1 (highest occupied molecular orbital) of the monofluoroanilines however, qualitatively correlate with the regioselectivity of the aromatic hydroxylation. Based on these results it is concluded that the cytochrome P-450 dependent aromatic hydroxylation of monofluoroanilines does not proceed by hydrogen or electron abstraction from the aniline substrate to give an aniline-NH. radical. The results rather suggest that cytochrome P-450 catalyzed aromatic hydroxylation of monofluoroanilines proceeds by an electrophilic attack of the (FeO)3+ species of cytochrome P-450 on a specific carbon atom of the aromatic aniline-ring.
Cnubben
N H
NH
Department of Biochemistry, Agricultural University, Wageningen, The Netherlands.
Vervoort
J
J
Veeger
C
C
Rietjens
I M
IM
D000814:Aniline Compounds / Q000378:metabolism* / Q000493:pharmacokinetics / Q000652:urine; D000818:Animals; D001711:Biotransformation; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D005461:Fluorine; D005464:Fluorobenzenes / Q000378:metabolism / Q000493:pharmacokinetics / Q000652:urine; D006900:Hydroxylation; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism; D008956:Models, Chemical; D051381:Rats; D017208:Rats, Wistar; D013237:Stereoisomerism
D000814:Aniline Compounds; D005464:Fluorobenzenes; D005461:Fluorine; C066212:2-fluoroaniline; C079598:3-fluoroaniline; C043455:4-fluoroaniline; D003577:Cytochrome P-450 Enzyme System