N-Acetyl-S-(N,N-diethylcarbamoyl) cysteine in rat nucleus accumbens, medial prefrontal cortex, and in rat and human plasma after disulfiram administration

Robert D. Winefield, Anthonius A.M. Heemskerk, Swetha Kaul, Todd D. Williams, Michael J. Caspers, Thomas E. Prisinzano, Elinore F. McCance-Katz, Craig E. Lunte, Morris D. Faiman

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Disulfiram (DSF), a treatment for alcohol use disorders, has shown some clinical effectiveness in treating addiction to cocaine, nicotine, and pathological gambling. The mechanism of action of DSF for treating these addictions is unclear but it is unlikely to involve the inhibition of liver aldehyde dehydrogenase (ALDH2). DSF is a pro-drug and forms a number of metabolites, one of which is N-acetyl-. S-(N,N-diethylcarbamoyl) cysteine (DETC-NAC). Here we describe a LCMS/MS method on a QQQ type instrument to quantify DETC-NAC in plasma and intracellular fluid from mammalian brain. An internal standard, the N,N-di-isopropylcarbamoyl homolog (MIM: 291. >. 128) is easily separable from DETC-NAC (MIM: 263. >. 100) on C18 RP media with a methanol gradient. The method's linear range is 0.5-500. nM from plasma and dialysate salt solution with all precisions better than 10% RSD. DETC-NAC and internal standards were recovered at better than 95% from all matrices, perchloric acid precipitation (plasma) or formic acid addition (salt) and is stable in plasma or salt at low pH for up to 24. h. Stability is observed through three freeze-thaw cycles per day for 7 days. No HPLC peak area matrix effect was greater than 10%. A human plasma sample from a prior analysis for S-(N,N-diethylcarbamoyl) glutathione (CARB) was found to have DETC NAC as well. In other human plasma samples from 62.5. mg/d and 250. mg/d dosing, CARB concentration peaks at 0.3 and 4. nM at 3. h followed by DETC-NAC peaks of 11 and 70. nM 2. h later. Employing microdialysis sampling, DETC-NAC levels in the nucleus accumbens (NAc), medial prefrontal cortex (mPFC), and plasma of rats treated with DSF reached 1.1, 2.5 and 80. nM at 6. h. The correlation between the appearance and long duration of DETC-NAC concentration in rat brain and the persistence of DSF-induced changes in neurotransmitters observed by Faiman et al. (Neuropharmacology, 2013, 75C, 95-105) is discussed.

Original languageEnglish
Pages (from-to)518-525
Number of pages8
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume107
DOIs
StatePublished - Mar 5 2015

Bibliographical note

Funding Information:
The Micromass Ultima was purchased with support from the KU Research Development Fund and the Acquity chromatograph with partial support from K-INBRE grant P20GM103418 to TDW. This work was supported in part by grants from the National Institute on Drug Abuse of the National Institute of Health to EFM grant numbers DA 023359 and DA 024982 and to MDF grant number DA 021727 .

Publisher Copyright:
© 2015 Elsevier B.V.

Keywords

  • Carbamathione
  • DETC-NAC
  • Disulfiram
  • Mercapturate pathway
  • Substance abuse disorders

ASJC Scopus subject areas

  • Analytical Chemistry
  • Pharmaceutical Science
  • Drug Discovery
  • Spectroscopy
  • Clinical Biochemistry

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