The demethylenation of methylenedioxymethamphetamine (“ecstasy”) by debrisoquine hydroxylase (CYP2D6)

doi:10.1016/0006-2952(94)90386-7

Biochemical Pharmacology

Volume 47, Issue 7, 29 March 1994, Pages 1151-1156

https://doi.org/10.1016/0006-2952(94)90386-7 Get rights and content

Abstract

The metabolism of methylenedioxymethamphetamine (MDMA, “ecstasy”) was examined in a microsomal preparation of the yeast Saccharomyces cerevisiae expressing human debrisoquine hydroxylase, CYP2D6. Only one product, dihydroxymethylamphetamine (DHMA), was detected in the incubation mixture, and this product accounted for all of the substrate consumption at low concentration (10μM). Mean ± SD values of apparent K_m(μM) and V_max (nmol/min per nmol P450) for the demethylenation of (+) and (−)-MDMA at low concentrations (1–1000 μM) were 1.72, 0.12 and 6.45, 0.10 and 2.90, 0.10 and 7.61, 0.06, respectively. At high concentrations (> 1000 μM) substrate inhibition was noted, with K_i values of 14.2 and 28.2 mM, respectively, for the (+) and (−) enantiomers. Incubation of MDMA isomers with human liver microsomes indicated that their demethylenation is deficient in the poor metabolizer phenotype. Thus, MDMA is converted to the catecholamine DHMA by CYP2D6, and this may give rise to genetically-determined differences in toxicity.

References (31)

G.R. Screaton et al.
Hyperpyrexia and rhabdomyolosis after MDMA (“ecstasy”) abuse
Lancet
(1992)
C.J. Schmidt et al.
Direct central effects of acute methylenedioxymethamphetamine on serotoningergic neurones
Eur J Pharmacol
(1988)
S.W. Ellis et al.
Catalytic activities of human debrisoquine 4-hydroxylase cytochrome P450 (CYP2D6) expressed in yeast
Biochem Pharmacol
(1992)
M.S. Ching et al.
The expression of cytochrome P450IA1 in the yeast Saccharomyces cerevisiae
Biochem Pharmacol
(1991)
A.G. Hildebrandt et al.
Hydrogen peroxide in hepatic microsomes
J.A. Dominic et al.
Depression of behaviour and the brain content of alpha-methylnorepinephrine and alpha-methyldopamine following the administration of alpha-methyldopa
Neuropharmacology
(1971)
R.A. Glennon et al.
The effect of MDMA (“Ecstasy”) and its optical isomers on schedule-controlled responding in mice
Pharmacol Biochem Behav
(1987)
R.E. Billings et al.
Aromatic hydroxylation of amphetamine with rat liver microsomes, perfused liver, and isolated hepatocytes
Biochem Pharmacol
(1978)
G.P. Dowling et al.
A report of five deaths associated with the use of MDEA and MDMA
J Am Med Assoc
(1987)
A.R. Winstock
Chronic paranoid psychosis after misuse of MDMA
Br Med J
(1991)

M. Hiramatsu et al.

Metabolism of methylenedioxymethamphetamine: formation of dihydroxymethylamphetamine and a quinone identified as its glutathione adduct

J Pharmacol Exp Ther

(1990)

Z. Zhao et al.

Synthesis and neurotoxicological evaluation of putative metabolites of the serotoninergic neurotoxin 2-(methylamino)-1-[3,4-(methylenedioxy)phenyl] propane [(methylenedioxy) methamphetamine]

Chem Res Toxicol

(1992)

L.Y. Lin et al.

Enzymatic and chemical demethylenation of (methylenedioxy)amphetamine (MDA) and (methylenedioxy)methamphetamine (MDMA) by rat brain microsomes

Chem Res Toxicol

(1992)

H.H. Maurer et al.

On the metabolism of 3,4-methylenedioxymethamphetamine (MDMA) in man

Ther Drug Monit

(1993)

Y. Kumagai et al.

Cytochrome P450 isozymes responsible for the metabolic activation of methylenedioxymethamphetamine (MDMA) in rat

FASEB J

(1992)

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Neurological and cognitive alterations induced by MDMA in humans
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Citation Excerpt :
There are a number of factors, both physiological and environmental, which could exacerbate the toxic effects of MDMA, through their interaction with individual differences in pharmacokinetics (Capela et al., 2009). For example, females are more likely to experience adverse effects, possibly due to the effects of hormones on pharmacokinetics (Liechti et al., 2001; Simmler et al., 2011) and various genetic polymorphisms can affect metabolism of both MDMA and serotonin (Hysek et al., 2012; Martín-Santos et al., 2010; Rietjens et al., 2012; Tucker et al., 1994), affecting overall neurological and cognitive alterations. In addition the environmental conditions when using ecstasy can exacerbate any adverse effect with higher ambient temperature (Capela et al., 2009; Green et al., 2003) and increased physical activity (Parrott et al., 2006) emerging as significant factors.
3,4 Methylenedioxymethamphetamine generally referred to as MDMA or ‘ecstasy’ is a ring-substituted phenethylamine stimulant which produces powerful empathogenic effects. Use of MDMA remains popular despite prohibition, and potential long-term negative consequences of repeated use. MDMA produces its acute subjective effects primarily by stimulating the release of serotonin via action at the serotonin transporter (SERT). There is evidence that MDMA administration may lead to long lasting neurotoxic effects on serotonin neurons in primates, and reductions in markers of central serotonin axons, and axon terminals in animals. In humans, demonstration of serotonergic neurotoxicity is much more difficult to identify, and much of the research is complicated by confounding issues of polysubstance use, genetic and environmental factors and reliance on self-reports of previous drug use. We do not review the mechanisms for neurotoxicity in detail as they are covered elsewhere in this special issue. There is a large body of literature, however, which has investigated potential cognitive and neurocognitive consequences of repeated MDMA use. Here we review the literature on cognition, and neuroimaging studies that have investigated structural and functional brain changes associated with ecstasy use.
Cognitive consequences of 3,4-methylenedioxymethamphetamine use
2020, Cognition and Addiction: A Researcher's Guide from Mechanisms Towards Interventions
This chapter summarizes our current knowledge on the prevalence of ecstasy use, the etiology of use, and possible psychological consequences resulting from using ecstasy. Drawing on both animal and human studies, the chapter discusses evidence for neurotoxicity of 3,4-methylenedioxymethamphetamine (MDMA)/ecstasy use and both structural and functional changes in human MDMA users. The chapter also reviews studies on cognitive function in human users of ecstasy and discusses methodological problems within this sample, namely polydrug use, drug purity, and premorbid functioning. There are a number of risk factors and individual differences in pharmacokinetic processing of the drug, which could make use more risky for certain subgroups of individuals. Combined with issues surrounding purity and strength, it is difficult to predict if an individual may be at risk of MDMA-related harm. Thus drug policymakers should seek to develop drug policy further to protect users of recreational drugs and provide harm reduction information.
Is toxicity of PMMA (paramethoxymethamphetamine) associated with cytochrome P450 pharmacogenetics?
2016, Forensic Science International
In 2010–2013, 29 fatal intoxications related to the designer drug paramethoxymethamphetamine (PMMA, 4-methoxymethamphetamine) occurred in Norway. The current knowledge about metabolism and toxicity of PMMA in humans is limited. Metabolism by the polymorphic cytochrome P450 (CYP) 2D6 enzyme to the psychoactive metabolite 4-hydroxymethamphetamine (OH-MA), and possibly by additional enzymes, is suggested to be involved in its toxicity. The aim of this work was to study the association between CYP genetics, PMMA metabolism and risk of fatal PMMA toxicity in humans. The frequency distribution of clinically relevant gene variants of CYP2D6, CYP2C9, CYP2C19 and CYP3A5, and the phenotypic blood CYP2D6 metabolic ratio (OH-MA/PMMA) in particular, were compared in fatal PMMA intoxications (n = 17) and nonfatal PMMA abuse controls (n = 30), using non-abusers (n = 305) as references for the expected genotype frequencies in the Norwegian population. Our study demonstrated that the CYP2D6 enzyme and genotype are important in the metabolism of PMMA to OH-MA in humans, but that other enzymes are also involved in this biotransformation. In the fatal PMMA intoxications, the blood concentrations of PMMA were higher and the CYP2D6 metabolic ratios were lower, than in the nonfatal PMMA abuse controls (median (range) 2.1 (0.03–5.0) vs 0.3 (0.1–0.9) mg/L, and ratio 0.6 (0.0–4.6) vs 2.1 (0.2–7.4) p = 0.021, respectively). Overall, our findings indicated that, in most cases, PMMA death occurred rapidly and at an early stage of PMMA metabolism, following the ingestion of large and toxic PMMA doses. We could not identify any genetic CYP2D6, CYP2C9, CYP2C19 or CYP3A5 predictive marker on fatal toxicity of PMMA in humans. The overrepresentation of the CYP2D6 poor metabolizer (PM) genotype found in the nonfatal PMMA abuse controls warrants further investigations.
Pharmacokinetics, pharmacodynamics and toxicology of new psychoactive substances (NPS): 2C-B, 4-fluoroamphetamine and benzofurans
2015, Drug and Alcohol Dependence
Citation Excerpt :
As a result, specific individuals with a disadvantageous genetic profile may be at higher risk for 2C-B-induced toxicity. Differences in human susceptibility to the toxicity of other amphetamine-like designer drugs has been reported previously (Carmo et al., 2004; Tucker et al., 1994; Colado et al., 1995; Jones and Simpson, 1999; Malpass et al., 1999). Furthermore, comparable 5- or 6-APB concentrations were detected in deceased individuals and in DUID cases, indicating the importance of interindividual differences in response to a similar exposure.
Recently, the number of new psychoactive substances (NPS) appearing on the illicit drug market has shown a marked increase. Although many users perceive the risk of using NPS as medium or low, these substances can pose a serious health risk and several NPS have been implicated in drug-related deaths. In Europe, frequently detected NPS are 4-bromo-2,5-dimethoxyphenethylamine (2C-B), 4-fluoroamphetamine (4-FA) and benzofurans (5-(2-aminopropyl)benzofuran (5-APB) or 6-(2-aminopropyl)benzofuran (6-APB)). However, little is known about the health risks of these specific NPS.
In this paper, existing literature on the pharmacokinetics and pharmacodynamics of 2C-B, 4-FA and benzofurans (5-APB/6-APB) was reviewed.
Our review showed that the clinical effects of 2C-B, 4-FA and benzofurans (5-APB/6-APB) are comparable with common illicit drugs like amphetamine and 3,4-methylenedioxymethamphetamine (MDMA). Therefore, NPS toxicity can be handled by existing treatment guidelines that are based on clinical effects instead of the specific drug involved. Even so, information on the health risks of these substances is limited to a number of case reports that are complicated by confounders such as analytical difficulties, mislabelling of drugs, concomitant exposures and interindividual differences.
To aid in early legislation, data on clinical effects from poisons centres and user fora should be combined with (in vitro) screening methods and collaboration on an (inter)national level is essential. As a result, potentially hazardous NPS could be detected more quickly, thereby protecting public health.
An in vitro approach to assessing a potential drug interaction between MDMA (ecstasy) and caffeine
2014, Toxicology in Vitro
3,4-Methylenedioxymethamphetamine (MDMA, ecstasy) is a popular recreational drug which causes long-term neurotoxicity and increased risk of fatality. In rats, MDMA toxicity is exacerbated by co-administration of caffeine. The aim of this study was to investigate whether caffeine altered the effects of MDMA in a battery of in vitro tests selected to model some of the known actions of MDMA in vivo. In cytotoxicity studies, caffeine modestly enhanced the effect of MDMA on neuronal N2a cell viability but not that of liver, intestinal or kidney cells. MDMA inhibited the formation of fluorescent metabolites by CYP2D6 ≫ CYP3A4 > CYP1A2 but this was not altered by caffeine. Similarly, the inhibition of synaptosomal [³H] 5-HT uptake by MDMA was not affected by the presence of caffeine. Thus, these in vitro tests failed to detect any substantial interaction between caffeine and MDMA, highlighting the difficulty of modelling in vivo drug interactions using in vitro tests. However, the results show that the inhibition of synaptosomal [³H] 5-HT uptake by MDMA was greater at 41 °C and 25 °C than at 37 °C which raises the possibility that MDMA’s effect on SERT in vivo may be increased as body temperature increases, contributing to its harmful effects in users.
Dioxygenases catalyzeo-demethylation and O,O-demethylenation with widespread roles in benzylisoquinoline alkaloid metabolism in opium poppy
2013, Journal of Biological Chemistry
Thebaine 6-O-demethylase (T6ODM) and codeine O-demethylase (CODM) catalyze late steps of morphine biosynthesis in opium poppy.
Results: New dealkylation reactions, including O,O-demethylenation, have been detected for T6ODM, CODM, and other 2-oxoglutarate/Fe(II)-dependent dioxygenases (ODDs). Virus-induced gene silencing supports physiological functions.
Conclusion: Certain ODDs are multifunctional dealkylating enzymes with widespread roles in benzylisoquinoline alkaloid metabolism.
Significance: Enzymes responsible for O-demethylation and O,O-demethylenation in plant alkaloid biosynthesis have been discovered.
In opium poppy, the antepenultimate and final steps in morphine biosynthesis are catalyzed by the 2-oxoglutarate/Fe(II)-dependent dioxygenases, thebaine 6-O-demethylase (T6ODM) and codeine O-demethylase (CODM). Further investigation into the biochemical functions of CODM and T6ODM revealed extensive and unexpected roles for such enzymes in the metabolism of protopine, benzo[c]phenanthridine, and rhoeadine alkaloids. When assayed with a wide range of benzylisoquinoline alkaloids, CODM, T6ODM, and the functionally unassigned paralog DIOX2, renamed protopine O-dealkylase, showed novel and efficient dealkylation activities, including regio- and substrate-specific O-demethylation and O,O-demethylenation. Enzymes catalyzing O,O-demethylenation, which cleave a methylenedioxy bridge leaving two hydroxyl groups, have previously not been reported in plants. Similar cleavage of methylenedioxy bridges on substituted amphetamines is catalyzed by heme-dependent cytochromes P450 in mammals. Preferred substrates for O,O-demethylenation by CODM and protopine O-dealkylase were protopine alkaloids that serve as intermediates in the biosynthesis of benzo[c]phenanthridine and rhoeadine derivatives. Virus-induced gene silencing used to suppress the abundance of CODM and/or T6ODM transcripts indicated a direct physiological role for these enzymes in the metabolism of protopine alkaloids, and they revealed their indirect involvement in the formation of the antimicrobial benzo[c]phenanthridine sanguinarine and certain rhoeadine alkaloids in opium poppy.

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Biochemical Pharmacology

Abstract

References (31)

Hyperpyrexia and rhabdomyolosis after MDMA (“ecstasy”) abuse

Lancet

Direct central effects of acute methylenedioxymethamphetamine on serotoningergic neurones

Eur J Pharmacol

Catalytic activities of human debrisoquine 4-hydroxylase cytochrome P450 (CYP2D6) expressed in yeast

Biochem Pharmacol

The expression of cytochrome P450IA1 in the yeast Saccharomyces cerevisiae

Biochem Pharmacol

Hydrogen peroxide in hepatic microsomes

Depression of behaviour and the brain content of alpha-methylnorepinephrine and alpha-methyldopamine following the administration of alpha-methyldopa

Neuropharmacology

The effect of MDMA (“Ecstasy”) and its optical isomers on schedule-controlled responding in mice

Pharmacol Biochem Behav

Aromatic hydroxylation of amphetamine with rat liver microsomes, perfused liver, and isolated hepatocytes

Biochem Pharmacol

A report of five deaths associated with the use of MDEA and MDMA

J Am Med Assoc

Chronic paranoid psychosis after misuse of MDMA

Br Med J

Metabolism of methylenedioxymethamphetamine: formation of dihydroxymethylamphetamine and a quinone identified as its glutathione adduct

J Pharmacol Exp Ther

Synthesis and neurotoxicological evaluation of putative metabolites of the serotoninergic neurotoxin 2-(methylamino)-1-[3,4-(methylenedioxy)phenyl] propane [(methylenedioxy) methamphetamine]

Chem Res Toxicol

Enzymatic and chemical demethylenation of (methylenedioxy)amphetamine (MDA) and (methylenedioxy)methamphetamine (MDMA) by rat brain microsomes

Chem Res Toxicol

On the metabolism of 3,4-methylenedioxymethamphetamine (MDMA) in man

Ther Drug Monit

Cytochrome P450 isozymes responsible for the metabolic activation of methylenedioxymethamphetamine (MDMA) in rat

FASEB J

Cited by (231)

Neurological and cognitive alterations induced by MDMA in humans

Cognitive consequences of 3,4-methylenedioxymethamphetamine use

Is toxicity of PMMA (paramethoxymethamphetamine) associated with cytochrome P450 pharmacogenetics?

Pharmacokinetics, pharmacodynamics and toxicology of new psychoactive substances (NPS): 2C-B, 4-fluoroamphetamine and benzofurans

An in vitro approach to assessing a potential drug interaction between MDMA (ecstasy) and caffeine

Dioxygenases catalyzeo-demethylation and O,O-demethylenation with widespread roles in benzylisoquinoline alkaloid metabolism in opium poppy