Comparison of the metabolism of mephedrone in cultured and in freshly isolated primary rat hepatocytes

Research output: Contribution to journalMeeting abstract

Abstract

Mephedrone (4-MMC) is a drug of abuse, which was controlled in 2010, and has been associated with several fatalities among users. In this study, the in vitro metabolism of 4-MMC in cultured Sprague-Dawley rat hepatocytes was characterised and Phase I and II metabolites quantified after specific periods of time in culture - 6, 24, and 48 hours. The cells were extracted and metabolites analysed using zwitterionic hydrophilic interaction (ZIC(r)-pHILIC) column and an exactive high resolution mass spectrometer instrument. Metabolism of 4-MMC yielded in 14 metabolites. These metabolites were structurally characterised on the basis of accurate mass analyses having been previously characterized by LC-MSn. The major identified metabolic routes for 4-MMC were found to be (i) 4’-methyl group oxidation and (ii) b-keto group reduction. The metabolism was compared to the published data of the mephedrone metabolism in freshly isolated rat hepatocytes. Three metabolites were not identified in culture compared to the previously published metabolism data in freshly isolated cells. Metabolite 4-carboxy-mephedrone was found to be the most prominent metabolite after 6 and 24 hours incubation, whereas nor-mephedrone was the predominant metabolite after 48 hours incubation. These data suggest that primary cultures of rat hepatocytes can be used to screen for metabolites of drugs like mephedrone to find out if the toxicity observed in users is related to formation of reactive metabolites and that the hepatocytes maintain their metabolic competency after 48 hours in culture.
Original languageEnglish
Pages (from-to)241-241
Number of pages1
JournalApplied in Vitro Toxicology
Volume2
Issue number4
DOIs
Publication statusPublished - 31 Dec 2016

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Metabolites
Metabolism
Rats
Hepatocytes
Street Drugs
Hydrophobic and Hydrophilic Interactions
Sprague Dawley Rats
mephedrone
Mass spectrometers
Pharmaceutical Preparations
Toxicity
Oxidation

Keywords

  • mephedrone
  • in vitro metabolism
  • rat hepatocytes
  • metabolism data

Cite this

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title = "Comparison of the metabolism of mephedrone in cultured and in freshly isolated primary rat hepatocytes",
abstract = "Mephedrone (4-MMC) is a drug of abuse, which was controlled in 2010, and has been associated with several fatalities among users. In this study, the in vitro metabolism of 4-MMC in cultured Sprague-Dawley rat hepatocytes was characterised and Phase I and II metabolites quantified after specific periods of time in culture - 6, 24, and 48 hours. The cells were extracted and metabolites analysed using zwitterionic hydrophilic interaction (ZIC(r)-pHILIC) column and an exactive high resolution mass spectrometer instrument. Metabolism of 4-MMC yielded in 14 metabolites. These metabolites were structurally characterised on the basis of accurate mass analyses having been previously characterized by LC-MSn. The major identified metabolic routes for 4-MMC were found to be (i) 4’-methyl group oxidation and (ii) b-keto group reduction. The metabolism was compared to the published data of the mephedrone metabolism in freshly isolated rat hepatocytes. Three metabolites were not identified in culture compared to the previously published metabolism data in freshly isolated cells. Metabolite 4-carboxy-mephedrone was found to be the most prominent metabolite after 6 and 24 hours incubation, whereas nor-mephedrone was the predominant metabolite after 48 hours incubation. These data suggest that primary cultures of rat hepatocytes can be used to screen for metabolites of drugs like mephedrone to find out if the toxicity observed in users is related to formation of reactive metabolites and that the hepatocytes maintain their metabolic competency after 48 hours in culture.",
keywords = "mephedrone, in vitro metabolism, rat hepatocytes, metabolism data",
author = "Ibrahim Alanazi and Grant, {M. Helen} and David Watson and Catherine Henderson",
year = "2016",
month = "12",
day = "31",
doi = "10.1089/aivt.2016.29007.abstracts",
language = "English",
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journal = "Applied in Vitro Toxicology",
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TY - JOUR

T1 - Comparison of the metabolism of mephedrone in cultured and in freshly isolated primary rat hepatocytes

AU - Alanazi, Ibrahim

AU - Grant, M. Helen

AU - Watson, David

AU - Henderson, Catherine

PY - 2016/12/31

Y1 - 2016/12/31

N2 - Mephedrone (4-MMC) is a drug of abuse, which was controlled in 2010, and has been associated with several fatalities among users. In this study, the in vitro metabolism of 4-MMC in cultured Sprague-Dawley rat hepatocytes was characterised and Phase I and II metabolites quantified after specific periods of time in culture - 6, 24, and 48 hours. The cells were extracted and metabolites analysed using zwitterionic hydrophilic interaction (ZIC(r)-pHILIC) column and an exactive high resolution mass spectrometer instrument. Metabolism of 4-MMC yielded in 14 metabolites. These metabolites were structurally characterised on the basis of accurate mass analyses having been previously characterized by LC-MSn. The major identified metabolic routes for 4-MMC were found to be (i) 4’-methyl group oxidation and (ii) b-keto group reduction. The metabolism was compared to the published data of the mephedrone metabolism in freshly isolated rat hepatocytes. Three metabolites were not identified in culture compared to the previously published metabolism data in freshly isolated cells. Metabolite 4-carboxy-mephedrone was found to be the most prominent metabolite after 6 and 24 hours incubation, whereas nor-mephedrone was the predominant metabolite after 48 hours incubation. These data suggest that primary cultures of rat hepatocytes can be used to screen for metabolites of drugs like mephedrone to find out if the toxicity observed in users is related to formation of reactive metabolites and that the hepatocytes maintain their metabolic competency after 48 hours in culture.

AB - Mephedrone (4-MMC) is a drug of abuse, which was controlled in 2010, and has been associated with several fatalities among users. In this study, the in vitro metabolism of 4-MMC in cultured Sprague-Dawley rat hepatocytes was characterised and Phase I and II metabolites quantified after specific periods of time in culture - 6, 24, and 48 hours. The cells were extracted and metabolites analysed using zwitterionic hydrophilic interaction (ZIC(r)-pHILIC) column and an exactive high resolution mass spectrometer instrument. Metabolism of 4-MMC yielded in 14 metabolites. These metabolites were structurally characterised on the basis of accurate mass analyses having been previously characterized by LC-MSn. The major identified metabolic routes for 4-MMC were found to be (i) 4’-methyl group oxidation and (ii) b-keto group reduction. The metabolism was compared to the published data of the mephedrone metabolism in freshly isolated rat hepatocytes. Three metabolites were not identified in culture compared to the previously published metabolism data in freshly isolated cells. Metabolite 4-carboxy-mephedrone was found to be the most prominent metabolite after 6 and 24 hours incubation, whereas nor-mephedrone was the predominant metabolite after 48 hours incubation. These data suggest that primary cultures of rat hepatocytes can be used to screen for metabolites of drugs like mephedrone to find out if the toxicity observed in users is related to formation of reactive metabolites and that the hepatocytes maintain their metabolic competency after 48 hours in culture.

KW - mephedrone

KW - in vitro metabolism

KW - rat hepatocytes

KW - metabolism data

U2 - 10.1089/aivt.2016.29007.abstracts

DO - 10.1089/aivt.2016.29007.abstracts

M3 - Meeting abstract

VL - 2

SP - 241

EP - 241

JO - Applied in Vitro Toxicology

JF - Applied in Vitro Toxicology

SN - 2332-1512

IS - 4

ER -