This is a proposal to continue our studies of the action of disulfiram. Disulfiram is currently the only drug which is approved in the United States for aversion therapy in the treatment of alcoholism. This drug interferes with the metabolism of ethanol by the inhibition of the enzyme aldehyde dehydrogenase (ALDH). The precise mechanism of this effect is related to the formation of one or more active metabolites of disulfiram. The purpose of this study is to examine the pharmacology of disulfiram with respect to the formation of active metabolites, including S-methyl-diethylthiocarbamate sulfoxide, which has been proposed by others as a new therapeutic agent in the treatment of alcoholism. We will examine the inhibition of ALDH by metabolites, as this enzyme is involved in the clinical effect of disulfiram. Mass spectrometry will be used to identify active metabolites as well as to determine how and where inhibitors interact with ALDH. We will also investigate whether free radical formation is involved in the metabolism of disulfiram as this mechanism has implications both for the action as well as for the toxicities of disulfiram. Studies will be performed in vitro, as well as in rats and in humans. We will also use expressed recombinante human ALDH's. The studies described in this proposal will shed new light on the mechanism of action of disulfiram and the pharmacology of its metabolites, more precisely define how aldehyde dehydrogenase is inhibited, and lead to a better understanding of new compounds for the treatment of alcoholism.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project (R01)
Project #
5R01AA009543-06
Application #
2837275
Study Section
Special Emphasis Panel (ZRG4-ALTX-1 (01))
Program Officer
Foudin, Laurie L
Project Start
1992-09-30
Project End
2000-11-30
Budget Start
1998-12-01
Budget End
2000-11-30
Support Year
6
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Mayo Clinic, Rochester
Department
Type
DUNS #
City
Rochester
State
MN
Country
United States
Zip Code
55905
Shen, M L; Johnson, K L; Mays, D C et al. (2001) Determination of in vivo adducts of disulfiram with mitochondrial aldehyde dehydrogenase. Biochem Pharmacol 61:537-45
Shen, M L; Benson, L M; Johnson, K L et al. (2001) Effect of enzyme inhibitors on protein quaternary structure determined by on-line size exclusion chromatography-microelectrospray ionization mass spectrometry. J Am Soc Mass Spectrom 12:97-104
Lipsky, J J; Shen, M L; Naylor, S (2001) In vivo inhibition of aldehyde dehydrogenase by disulfiram. Chem Biol Interact 130-132:93-102
Pike, M G; Mays, D C; Macomber, D W et al. (2001) Metabolism of a disulfiram metabolite, S-methyl N,N-diethyldithiocarbamate, by flavin monooxygenase in human renal microsomes. Drug Metab Dispos 29:127-32
Shen, M L; Lipsky, J J; Naylor, S (2000) Role of disulfiram in the in vitro inhibition of rat liver mitochondrial aldehyde dehydrogenase. Biochem Pharmacol 60:947-53
Shen, M L; Johnson, K L; Mays, D C et al. (2000) Identification of the protein-drug adduct formed between aldehyde dehydrogenase and S-methyl-N,N-diethylthiocarbamoyl sulfoxide by on-line proteolytic digestion high performance liquid chromatography electrospray ionization mass spectrometry. Rapid Commun Mass Spectrom 14:918-23
Kathmann, E C; Naylor, S; Lipsky, J J (2000) Rat liver constitutive and phenobarbital-inducible cytosolic aldehyde dehydrogenases are highly homologous proteins that function as distinct isozymes. Biochemistry 39:11170-6
Pike, M G; Martin, Y N; Mays, D C et al. (1999) Roles of FMO and CYP450 in the metabolism in human liver microsomes of S-methyl-N,N-diethyldithiocarbamate, a disulfiram metabolite. Alcohol Clin Exp Res 23:1173-9
Kathmann, E C; Lipsky, J J (1999) Cloning and expression of a cDNA encoding a constitutively expressed rat liver cytosolic aldehyde dehydrogenase. Adv Exp Med Biol 463:237-41
Mays, D C; Tomlinson, A J; Johnson, K L et al. (1999) Inhibition of human mitochondrial aldehyde dehydrogenase by metabolites of disulfiram and structural characterization of the enzyme adduct by HPLC-tandem mass spectrometry. Adv Exp Med Biol 463:61-70

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