Characterization of moclobemide N-oxidation in human liver microsomes

被引:13
作者
Hoskins, J
Shenfield, G
Murray, M
Gross, A
机构
[1] Royal N Shore Hosp, Dept Clin Pharmacol, St Leonards, NSW 2065, Australia
[2] Univ New S Wales, Sch Physiol & Pharmacol, Sydney, NSW 2052, Australia
关键词
D O I
10.1080/00498250110055488
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
1. Moclobemide undergoes morpholine ring N-oxidation to form a major metabolite in plasma, Ro12-5637. 2. The kinetics of moclobemide N-oxidation in human liver microsomes (HLM). (n = 6) have been investigated and the mixed-function oxidase enzymes catalysing this reaction have been identified using inhibition, enzyme correlation, altered pH and heat pretreatment experiments. 3. N-oxidation followed single enzyme Michealis-Menten kinetics (0.02-4.0 mM). K-m (app) and V-max ranged from 0.48 to 1.35 mM (mean +/- SD 0.77 +/- 0.34 mM) and 0.22 to 2.15 nmol mg(-1) min(-1) (1.39 +/- 0.80 nmol mg(-1) min(-1)), respectively. 4. The N-oxidation of moclobemide strongly correlated with benzydamine N-oxidation, a probe reaction for flavin-containing monooxygenase (FMO) activity, (0.1 mM moclobemide, r(S) = 0.81; p < 0.005; 4 mM moclobemide, r(S) = 0.94; p = 0.0001). Correlations were observed between moclobemide N-oxidation and specific cytochrome P450 (CYP) activities at both moclobemide concentrations (0.1 mM moclobemide, CYP2C19 r(S) = 0.66; p < 0.05; 4 mM moclobemide, CYP2E1 r(S) = 0.56; p < 0.05). 5. The general P450 inhibitor, N-benzylimidazole, did not affect the rate of Ro12-5637 formation (0% inhibition versus control) at 1.3 mM moclobemide. Furthermore, the rate of Ro12-5637 formation in HLM was unaffected by inhibitors or substrates of specific P450s (< 10% inhibition versus control). 6. Heat pretreatment of HLM in the absence of NADPH (inactivating FMOs) resulted in 97% inhibition of Ro12-5637 formation. N-oxidation activity was greatest when incubated at pH 8.5. These results are consistent with the reaction being FMO mediated. 7. In conclusion, moclobemide N-oxidation activity has been observed in HLM in vitro and the reaction is predominantly catalysed by FMOs with a potentially small contribution from cytochrome P450 isoforms.
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页码:387 / 397
页数:11
相关论文
共 29 条
[1]   DETERMINATION OF BENZYDAMINE AND ITS N-OXIDE IN BIOLOGICAL-FLUIDS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY [J].
BALDOCK, GA ;
BRODIE, RR ;
CHASSEAUD, LF ;
TAYLOR, T .
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1990, 529 (01) :113-123
[2]  
Bourrie M, 1996, J PHARMACOL EXP THER, V277, P321
[3]  
Brunelle A, 1997, DRUG METAB DISPOS, V25, P1001
[4]  
Cashman JR, 2000, DRUG METAB DISPOS, V28, P169
[5]  
CASHMAN JR, 1993, DRUG METAB DISPOS, V21, P492
[6]  
Cashman JR, 1999, HANDBOOK OF DRUG METABOLISM, P477
[7]   EVALUATION OF TRIACETYLOLEANDOMYCIN, ALPHA-NAPHTHOFLAVONE AND DIETHYLDITHIOCARBAMATE AS SELECTIVE CHEMICAL PROBES FOR INHIBITION OF HUMAN CYTOCHROMES P450 [J].
CHANG, TKH ;
GONZALEZ, FJ ;
WAXMAN, DJ .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1994, 311 (02) :437-442
[8]   DEVELOPMENT AND PRELIMINARY APPLICATION OF A SIMPLE ASSAY OF S-MEPHENYTOIN 4-HYDROXYLASE ACTIVITY IN HUMAN LIVER-MICROSOMES [J].
CHIBA, K ;
MANABE, K ;
KOBAYASHI, K ;
TAKAYAMA, Y ;
TANI, M ;
ISHIZAKI, T .
EUROPEAN JOURNAL OF CLINICAL PHARMACOLOGY, 1993, 44 (06) :559-562
[9]  
DAPRADA M, 1989, J PHARMACOL EXP THER, V248, P400
[10]   Missense mutation in flavin-containing mono-oxygenase 3 gene, FMO3, underlies fish-odour syndrome [J].
Dolphin, CT ;
Janmohamed, A ;
Smith, RL ;
Shephard, EA ;
Phillips, IR .
NATURE GENETICS, 1997, 17 (04) :491-494