Isolation of N-vinylprotoporphyrin IX after hepatic cytochrome P450 inactivation by 3-[(arylthio)ethyl]sydnone in chick embryos pretreated with phenobarbital, glutethimide, dexamethasone, and beta-naphthoflavone: differential inhibition of ferrochelatase by N-vinylprotoporphyrin regioisomers.
Several xenobiotics caused hepatic porphyrin accumulation through mechanism-based inactivation of cytochrome P450(P450) and heme alkylation. Loss of iron from the alkylated heme results in formation of an N-alkylporphyrin, which is a potent inhibitor of ferrochelatase. N-Vinylprotoporphyrin IX (N-vinylPP) was identified in chick embryo liver after in ovo administration of 3-[(arylthio)ethyl]sydnone (TTMS). Pretreatment of chick embryos with beta-naphthoflavone, which causes a 90-fold increase in P450 1A levels, did not increase the formation of N-vinylPP after TTMS administration, showing that the heme moiety of P450 1A does not contribute to the formation of N-vinylPP. Increased amounts of N-vinylPP were isolated from dexamethasone-, phenobarbital-, and glutethimide-pretreated chick embryos, and it is possible that P450 2H and/or a P450 3A-like isozyme contributes to the formation of N-vinylPP. The ring B-substituted (NB) regioisomer of N-vinylPP constituted the lowest percentage of the total regioisomers (9-13%) in untreated and drug-induced chick embryos, thus supporting the concept that ring B of heme is occluded by a protein residue in the P450 active site. Previously, the finding that the NB regioisomer of N-ethylprotoporphyrin IX had one fifth the potency of the ring A-substituted (NA) regioisomer as a ferrochelatase inhibitor led to a proposal that an A-C ring tilt was important in N-alkylprotoporphyrins for ferrochelatase inhibition. The finding in the present study that the NA and NB regioisomers of N-vinylPP are equipotent does not support the above proposal. The ring C-substituted (NC) and ring D-substituted (ND) regioisomers of N-vinylPP had low potency.
Kimmett
S M
SM
Department of Pharmacology, Queen's University, Kingston, Ontario, Canada.
Whitney
R A
RA
Marks
G S
GS
D000818:Animals; D001571:Benzoflavones / Q000494:pharmacology*; D002642:Chick Embryo; D065607:Cytochrome P-450 Enzyme Inhibitors; D003907:Dexamethasone / Q000494:pharmacology*; D005294:Ferrochelatase / Q000037:antagonists & inhibitors*; D005984:Glutethimide / Q000494:pharmacology*; D008099:Liver / Q000378:metabolism; D010634:Phenobarbital / Q000494:pharmacology*; D011524:Protoporphyrins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology*; D013237:Stereoisomerism; D013558:Sydnones / Q000494:pharmacology*; D019324:beta-Naphthoflavone
D001571:Benzoflavones; D065607:Cytochrome P-450 Enzyme Inhibitors; D011524:Protoporphyrins; D013558:Sydnones; C048192:3-(2-(2,4,6-trimethylphenyl)thioethyl)-4-methylsydnone; D019324:beta-Naphthoflavone; D003907:Dexamethasone; D005984:Glutethimide; D005294:Ferrochelatase; D010634:Phenobarbital