title: Oxidative metabolism of bupivacaine into pipecolylxylidine in humans is mainly catalyzed by CYP3A.
abstract: Bupivacaine is used to provide prolonged anesthesia and postoperative analgesia. The human cytochrome P450 (CYP) involved in bupivacaine degradation into pipecolylxylidine (PPX), its major metabolite, has, to our knowledge, never been described. Microsome samples were prepared from six human livers and incubated in the presence of bupivacaine. The concentrations of PPX in the microsomal suspensions were assessed, and K(m) and V(max) values were calculated. Bupivacaine incubations were then performed with specific CYP substrates and inhibitors. For each sample of hepatic microsomes, the correlation between the rate of PPX formation and the corresponding erythromycin N-demethylase activity was analyzed. Finally, an immunoinhibition study using an anti-rabbit CYP3A6 antibody and assays with cDNA-expressed human CYP were conducted. The apparent K(m) and V(max) values of bupivacaine were, respectively, 125 microM and 4.78 nmol/min/mg of microsomal protein. The strongest inhibition of bupivacaine metabolism was obtained for troleandomycin (-95% at 50 microM), a specific CYP3A inhibitor. The correlation between PPX formation and erythromycin N-demethylase activity showed an R value of 0.99 whereas anti-rabbit CYP3A6 antibody inhibited the degradation of bupivacaine into PPX by 99%. Finally, CYP1A2 and CYP2E1 cDNA-expressed forms of human CYP did not allow PPX formation, CYP2C19 and CYP2D6 produced only small amounts whereas CYP3A4 most efficiently metabolized bupivacaine into PPX. These results demonstrated that bupivacaine degradation into PPX was mediated in humans by CYP3A.
authors.affiliation: Laboratory of Medical Pharmacology (EA2199), School of Medicine, School of Pharmacy, Université Méditerranée, Marseilles, France.
authors.forename: M
authors.initials: M
authors.lastname: Gantenbein
authors.forename: L
authors.initials: L
authors.lastname: Attolini
authors.forename: B
authors.initials: B
authors.lastname: Bruguerolle
authors.forename: P H
authors.initials: PH
authors.lastname: Villard
authors.forename: F
authors.initials: F
authors.lastname: Puyoou
authors.forename: A
authors.initials: A
authors.lastname: Durand
authors.forename: B
authors.initials: B
authors.lastname: Lacarelle
authors.forename: J
authors.initials: J
authors.lastname: Hardwigsen
authors.forename: Y P
authors.initials: YP
authors.lastname: Le-Treut
mesh_terms: D000779:Anesthetics, Local / Q000493:pharmacokinetics*; D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology; D001189:Aryl Hydrocarbon Hydroxylases; D002045:Bupivacaine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics*; D002478:Cells, Cultured; D051544:Cytochrome P-450 CYP3A; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D007313:Insecta; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism; D010084:Oxidation-Reduction; D010089:Oxidoreductases, N-Demethylating / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*
chemical_list: D000779:Anesthetics, Local; D019138:Antibodies, Blocking; D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; D051544:Cytochrome P-450 CYP3A; D010089:Oxidoreductases, N-Demethylating; C025149:2',6'-pipecoloxylidide; D002045:Bupivacaine