title: Influence of CYP2D6 polymorphism on 3,4-methylenedioxymethamphetamine ('Ecstasy') cytotoxicity.
abstract: OBJECTIVE
  Remarkable interindividual differences in 3,4-methylenedioxymethamphetamine ('Ecstasy')-mediated toxicity have been reported in humans. Therefore, we tested whether CYP2D6 or its variant alleles as well as CYP3A4 influence the susceptibility to 3,4-methylenedioxymethamphetamine.
  
  
  METHODS
  3,4-Methylenedioxymethamphetamine cytotoxicity was determined in V79 cells expressing human wild-type CYP2D6 (CYP2D6*1), the low-activity alleles CYP2D6*2, *9, *10, and *17, as well as human CYP3A4. Metabolites of 3,4-methylenedioxymethamphetamine formed by the different cell lines were quantified by high-performance liquid chromatography/electrochemical detector.
  
  
  RESULTS
  Toxicity of 3,4-methylenedioxymethamphetamine was clearly increased in cells expressing CYP2D6*1 compared with the parental cells devoid of CYP-dependent enzymatic activity. Toxicity in V79 CYP2D6*1 cells was also higher than in V79 cell lines expressing the low-activity alleles CYP2D6*2, *9, *10, or *17. In contrast to CYP2D6, the CYP3A4 isoenzyme did not enhance 3,4-methylenedioxymethamphetamine toxicity. Formation of the oxidative 3,4-methylenedioxymethamphetamine metabolite N-methyl-alpha-methyldopamine was greatly enhanced in V79 cell line transfected with CYP2D6*1 compared to all other cell lines. The increase in the cytotoxic effects of 3,4-methylenedioxymethamphetamine observed in this cell line was therefore suspected to be a consequence of the production of this metabolite. This was further investigated by testing the cytotoxicity of N-methyl-alpha-methyldopamine to the control cell line. The results confirmed our hypothesis as the metabolite proved to be more than 100-fold more toxic than the parent compound 3,4-methylenedioxymethamphetamine.
  
  
  CONCLUSIONS
  CYP2D6*1 mediates 3,4-methylenedioxymethamphetamine toxicity via formation of N-methyl-alpha-methyldopamine. Therefore, it will be important to investigate whether CYP2D6 ultrarapid metabolizers are overrepresented in the cases of 3,4-methylenedioxymethamphetamine intoxications.
authors.affiliation: REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Porto, Portugal. helenacarmo@ff.up.pt
authors.forename: Helena
authors.initials: H
authors.lastname: Carmo
authors.forename: Marc
authors.initials: M
authors.lastname: Brulport
authors.forename: Matthias
authors.initials: M
authors.lastname: Hermes
authors.forename: Franz
authors.initials: F
authors.lastname: Oesch
authors.forename: Renata
authors.initials: R
authors.lastname: Silva
authors.forename: Luísa M
authors.initials: LM
authors.lastname: Ferreira
authors.forename: Paula S
authors.initials: PS
authors.lastname: Branco
authors.forename: Douwe de
authors.initials: Dd
authors.lastname: Boer
authors.forename: Fernando
authors.initials: F
authors.lastname: Remião
authors.forename: Félix
authors.initials: F
authors.lastname: Carvalho
authors.forename: Michael R
authors.initials: MR
authors.lastname: Schön
authors.forename: Niels
authors.initials: N
authors.lastname: Krebsfaenger
authors.forename: Johannes
authors.initials: J
authors.lastname: Doehmer
authors.forename: Maria de Lourdes
authors.initials: Mde L
authors.lastname: Bastos
authors.forename: Jan G
authors.initials: JG
authors.lastname: Hengstler
mesh_terms: D000483:Alleles; D000818:Animals; D002384:Catalysis; D002460:Cell Line; D006224:Cricetinae; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics* / Q000378:metabolism; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D003846:Deoxyepinephrine / Q000031:analogs & derivatives / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006213:Hallucinogens / Q000494:pharmacology / Q000633:toxicity*; D008954:Models, Biological; D018817:N-Methyl-3,4-methylenedioxyamphetamine / Q000493:pharmacokinetics / Q000633:toxicity*; D010084:Oxidation-Reduction; D011110:Polymorphism, Genetic
chemical_list: D006213:Hallucinogens; C023492:alpha-methyldopamine; D003577:Cytochrome P-450 Enzyme System; C104464:CYP3A protein, human; D019389:Cytochrome P-450 CYP2D6; D051544:Cytochrome P-450 CYP3A; D018817:N-Methyl-3,4-methylenedioxyamphetamine; D003846:Deoxyepinephrine