DETERMINATION OF TOLBUTAMIDE HYDROXYLATION IN RAT-LIVER MICROSOMES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY - EFFECT OF PSYCHOACTIVE-DRUGS ON INVITRO ACTIVITY

被引:13
作者
HO, JW
MOODY, DE
机构
[1] Center for Human Toxicology, Department of Pharmacology and Toxicology, University of Utah, Salt Lake City
关键词
D O I
10.1016/0024-3205(93)90284-A
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
A simplified HPLC method for tolbutamide metabolism to hydroxytolbutamide has been used to screen sixty psychoactive drugs for their ability to inhibit rat liver microsomal tolbutamide hydroxylation. One-step extraction with diethyl ether was followed by reconstitution and isocratic HPLC analysis with a binary mobile phase (ammonium phosphate:methanol, 45:55, v/v). Nanogram amounts of hydroxytolbutamide formation were estimated with UV detection at 240 nm. Hydroxytolbutamide formation was linear with incubation times of 40 - 120 min, but specific activity increased with increases in microsomal protein (0.15 - 1.10 mg). A differential inhibitory response was demonstrated for tolbutamide and debrisoquine hydroxylation to 5 psychoactive drugs, suggesting that tolbutamide hydroxylation is not dependent on P4502D1. Sixty psychoactive drugs, or drug metabolites, (at 33 muM) were then co-incubated with tolbutamide (at 2.5 and 10.2 muM). Tolbutamide hydroxylation was refractory (<25% inhibition) to twenty-four of the drugs and only mildly inhibited (25-50% inhibition) by twenty-eight. Two compounds, trans-3-methylfentanyl and flurazepam, produced >50% inhibition that was independent of tolbutamide concentration. Five of the drugs (methadone, chlorpheniramine, meperidine, 6-monoacetylmorphine and methylphenidate), however, caused greater than 50% inhibition in a competitive manner which suggests these drugs may share an affinity for the substrate binding site for tolbutamide.
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页码:21 / 28
页数:8
相关论文
共 28 条
[1]   SELECTIVE-INHIBITION OF DRUG OXIDATION AFTER SIMULTANEOUS ADMINISTRATION OF 2 PROBE DRUGS, ANTIPYRINE AND TOLBUTAMIDE [J].
BACK, DJ ;
TJIA, J ;
MONIG, H ;
OHNHAUS, EE ;
PARK, BK .
EUROPEAN JOURNAL OF CLINICAL PHARMACOLOGY, 1988, 34 (02) :157-163
[2]  
BACK DJ, 1984, BR J CLIN PHARM, V18, P557
[3]   THE CHARACTERISTICS OF THE MICROSOMAL HYDROXYLATION OF TOLBUTAMIDE [J].
BELANGER, PM ;
STHILAIRE, S .
CANADIAN JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY, 1991, 69 (03) :400-405
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[5]   SIMPLE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD FOR THE DETERMINATION OF TOLBUTAMIDE AND ITS METABOLITES IN HUMAN-PLASMA AND URINE USING PHOTODIODE-ARRAY DETECTION [J].
CSILLAG, K ;
VERECZKEY, L ;
GACHALYI, B .
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1989, 490 (02) :355-363
[6]  
GEE SJ, 1984, DRUG METAB DISPOS, V12, P174
[7]   INVITRO INHIBITION OF A POLYMORPHIC HUMAN-LIVER P-450 ISOZYME BY NARCOTIC ANALGESICS [J].
HENTHORN, TK ;
SPINA, E ;
DUMONT, E ;
VONBAHR, C .
ANESTHESIOLOGY, 1989, 70 (02) :339-342
[8]   GAS-CHROMATOGRAPHY MASS-SPECTROMETRY ASSAYS FOR THE DETERMINATION OF DEBRISOQUINE AND SPARTEINE METABOLITES IN MICROSOMAL FRACTIONS OF RAT-LIVER [J].
HO, JW ;
MOODY, DE .
ANALYTICAL BIOCHEMISTRY, 1992, 203 (02) :348-351
[9]   SIMULTANEOUS DETERMINATION OF TOLBUTAMIDE AND ITS HYDROXY AND CARBOXY METABOLITES IN PLASMA AND URINE BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY [J].
KEAL, J ;
STOCKLEY, C ;
SOMOGYI, A .
JOURNAL OF CHROMATOGRAPHY, 1986, 378 (01) :237-241
[10]  
KNODELL RG, 1987, J PHARMACOL EXP THER, V241, P1112