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http://dmd.aspetjournals.org/content/suppl/2009/03/19/dmd.108.026203.DC1.html
0090-9556/09/3706-11521156$20.00
DRUG METABOLISM AND DISPOSITION
Copyright 2009 by The American Society for Pharmacology and Experimental Therapeutics
DMD 37:11521156, 2009
Short Communication
The Role of Human Hepatic Cytochrome P450 Isozymes in the
Metabolism of Racemic 3,4-Methylenedioxyethylamphetamine and
Its Single EnantiomersS
Received December 16, 2008; accepted March 12, 2009
ABSTRACT:
incubated using heterologously expressed human P450s, and
the corresponding metabolites dihydroxyethylamphetamine and
methylenedioxyamphetamine were determined by gas chromatography-mass spectrometry after chiral derivatization with S-heptafluorobutyrylprolyl chloride. The highest contributions to both
metabolic steps as calculated from the enzyme kinetic data were
obtained for CYP3A4 and CYP2D6 at substrate concentrations
corresponding to plasma concentrations of recreational users after intake of racemic MDEA. Both metabolic reactions were found
to be enantioselective with a general preference for the S-enantiomers, which was particularly pronounced in the case of CYP2C19.
In conclusion, different pharmacokinetic properties of MDEA enantiomers observed in vivo are therefore partially caused by P450dependent enantioselective metabolism.
1153
low-affinity component (Clarke, 1998). If eq. 2 was found to fit the data
significantly better (F test, P 0.05), biphasic kinetics were assumed.
V max [S]
K m [S]
(1)
V max,1 [S]
CLint,2 S]
K m,1 [S]
(2)
V
V
The relative activity factor approach (Crespi and Miller, 1999; Venkatakrishnan et al., 2000; Grime and Riley, 2006) was used to account for
differences in functional levels of redox partners between the two enzyme
sources.
The corrected activities (contributions), the percentages of net clearance by
a particular P450 at a certain substrate concentration, can be calculated
according to eq. 3:
clearanceenyzme[%]
contributionenyzme
100
contributionenzyme
(3)
FIG. 1. The two main metabolic steps of R- and S-MDEA leading to the formation of the corresponding enantiomers of dihydroxyethylamphetamine (DHEA) and MDA.
1154
MEYER ET AL.
TABLE 1
Kinetic data for the two main metabolic steps of (R,S-)MDEA
Units used are as follows: Vmax, picomoles per minute per picomole; and Km, micromolar.
MDA Formation
R,S-MDEA
R-MDEA
(racemic MDEA)
S-MDEA
(racemic MDEA)
S-MDEA
(single enantiomers)
Vmax
Km
Vmax/Km
Vmax
Km
Vmax/Km
19 0.6
23 0.8
22 0.9
3.3 0.4*
2.4 0.4*
10 0.4
12 0.5
10 0.5
3.2 0.2*
2.1 0.2*
9.0 0.4
11 0.7
12 0.4
2.2 0.2*
3.8 0.3*
13 0.5
18 1.2
15 0.4
2.5 0.5*
13 2.0*
16 0.8
19 0.8
8.7 1.5*
5.1 0.3
3.2 0.4*
91 11
124 17
110 17
4.6 1.9*
10 4.8*
65 11
82 13
126 6
6.5 0.6*
16 2.4*
32 6
53 15
31 19
11 0.2*
18 1.7*
179 21
115 34
187 19
3.6 2.7*
76 18*
126 21
99 20
1.8 0.9*
20 5
7.3 2.5*
0.21
0.19
0.20
0.72
0.24
0.15
0.15
0.08
0.49
0.13
0.28
0.21
0.39
0.20
0.21
0.07
0.16
0.08
0.69
0.17
0.13
0.19
4.83
0.26
0.44
ND
ND
3.0 0.1
9.2 0.5
0.8 0.1*
ND
ND
ND
4.4 0.3
0.4 0.2*
ND
ND
2.9 0.3
4.8 0.3
0.3 0.08*
ND
ND
2.4 0.1
9.3 0.2
1.1 0.08*
ND
ND
3.3 0.1
9.5 0.1
0.7 0.2*
ND
ND
54 7
2.8 0.9
45 16*
ND
ND
ND
2.0 0.6
21 5*
ND
ND
25 8
1.0 0.3
13 4*
ND
ND
139 20
1.4 0.2
38 7*
ND
ND
18 2.8
0.7 0.07
36 8*
ND
ND
0.06
3.29
0.02
ND
ND
ND
2.20
0.02
ND
ND
0.12
4.80
0.02
ND
ND
0.02
6.64
0.03
ND
ND
0.18
13.57
0.02
R-MDEA
(single enantiomers)
CYP1A2
CYP2B6
CYP2C19
CYP2D6
CYP3A4
CYP1A2
CYP2B6
CYP2C19
CYP2D6
CYP3A4
CYP1A2
CYP2B6
CYP2C19
CYP2D6
CYP3A4
CYP1A2
CYP2B6
CYP2C19
CYP2D6
CYP3A4
CYP1A2
CYP2B6
CYP2C19
CYP2D6
CYP3A4
DHEA Formation
and hence their concentrations must be much higher as in the respective incubations of the racemate to reach saturation.
CYP2D6 turned out to be the isozyme with the highest affinity
toward racemic MDEA and its enantiomers (Table 1). In addition,
there is a generally higher affinity toward S-MDEA than R-MDEA
concerning all involved isozymes. The obvious difference in the Km
values of racemic MDEA and the respective enantiomers might be
caused by interactions of R- and S-MDEA in incubations of the
FIG. 2. Plots of calculated total net clearance (in %) versus substrate concentration
for the N-deethylation and demethylenation.
1155
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MEYER ET AL.
References
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Address correspondence to: Dr. Hans H. Maurer, Department of Experimental and Clinical Toxicology, Institute of Experimental and Clinical Pharmacology
and Toxicology, Saarland University, Building 46, D-66421 Homburg (Saar), Germany. E-mail: hans.maurer@uks.eu
MARKUS R. MEYER
Department of Experimental and
FRANK T. PETERS
Clinical Toxicology, Institute of Experimental
HANS H. MAURER
and Clinical Pharmacology and Toxicology,
Saarland University, Homburg (Saar), Germany