title: Inhibition of recombinant human mitochondrial aldehyde dehydrogenase by two intermediate metabolites of disulfiram.
abstract: Disulfiram is used in aversion therapy for alcoholism. S-Methyl-N,N-diethylthiocarbamate (MeDTC) sulfoxide, a potent inhibitor of the target enzyme mitochondrial aldehyde dehydrogenase (ALDH2), is thought to be the principal active metabolite of disulfiram in vivo. We examined the effects on recombinant human ALDH2 of two intermediate metabolites of disulfiram, S-methyl-N,N-diethyldithiocarbamate (MeDDC) sulfoxide and MeDDC sulfine. MeDDC sulfoxide was a potent inhibitor of ALDH2 with an IC50 of 2.2 +/- 0.5 microM (mean +/- SD, N = 4) after preincubation with enzyme for 30 min. MeDDC sulfine was a relatively weak inhibitor of ALDH2 under the same conditions with an IC50 value of 62 +/- 14 microM. The inhibition of ALDH2 by both compounds was irreversible and did not require the cofactor NAD. The latter finding demonstrates that inactivation of ALDH2 is independent of the dehydrogenase activity of the enzyme. GSH blocked almost completely the inhibition by 20 microM of MeDDC sulfoxide and greatly diminished the inhibition by 200 microM of MeDDC sulfine. Inactivation by MeDDC sulfoxide was time dependent. MeDTC sulfoxide was a more potent inhibitor of recombinant human ALDH2 (IC50 = 1.4 +/- 0.3 microM after preincubation for 15 min) than either of the intermediate metabolites, and its inhibition was unaffected by GSH. Our results suggest that these newer intermediate metabolites of disulfiram, especially the more potent MeDTC sulfoxide, have the potential to inhibit the target enzyme ALDH2 in patients receiving disulfiram. However, until the significance of the interactions of the inhibitors with GSH is more fully understood, the contribution of MeDDC sulfine and MeDDC sulfoxide to the pharmacological effects of disulfiram in vivo is uncertain.
authors.affiliation: Department of Pharmacology, Mayo Medical School, Mayo Clinic/Foundation, Rochester, MN 55905, USA. mays.dennis@mayo.edu
authors.forename: D C
authors.initials: DC
authors.lastname: Mays
authors.forename: P
authors.initials: P
authors.lastname: Ortiz-Bermudez
authors.forename: J P
authors.initials: JP
authors.lastname: Lam
authors.forename: I H
authors.initials: IH
authors.lastname: Tong
authors.forename: A H
authors.initials: AH
authors.lastname: Fauq
authors.forename: J J
authors.initials: JJ
authors.lastname: Lipsky
mesh_terms: D000427:Alcohol Deterrents / Q000493:pharmacokinetics / Q000494:pharmacology*; D000444:Aldehyde Dehydrogenase / Q000037:antagonists & inhibitors*; D004221:Disulfiram / Q000493:pharmacokinetics / Q000494:pharmacology*; D004050:Ditiocarb / Q000031:analogs & derivatives* / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D000427:Alcohol Deterrents; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; C112317:S-methyl-N,N-diethyldithiocarbamate sulfine; C073220:S-methyl N,N-diethylthiolcarbamate sulfoxide; D004050:Ditiocarb; D000444:Aldehyde Dehydrogenase; D004221:Disulfiram