ABSTRACT
The role of individual rat liver cytochrome P-450 isozymes in the metabolism of the skeletal muscle relaxant, dantrolene, was studied. Following incubation of dantrolene with hepatic microsomes from 3-methylcholanthrene-treated rats, two major hydroxylated metabolites were identified. Using inhibitory antibodies specific for individual cytochrome P-450 isozymes, cytochromes P-450 1A1, 1A2, and 3A were identified to be involved in dantrolene hydroxylations. In liver microsomes from 3-methylcholanthrene-treated rats, antibodies specific for cytochrome P-450 1A1 and 1A2 inhibited hydroxylation of dantrolene by 60% and 20%, respectively. Kinetics studies using these microsomes showed that dantrolene hydroxylation was biphasic with a low KM (0.06-0.08 microM) and high KM (5-7 microM). Cytochrome P-450 1A1 was responsible for the low KM hydroxylation of dantrolene, whereas cytochrome P-450 1A2 was responsible for the high KM. In hepatic microsomes from pregnenolone-16 alpha-carbonitrile-treated rats, an antibody specific for cytochrome P-450 3A completely inhibited the formation of 5-hydroxydantrolene, the major metabolite formed by these microsomes.
Subject(s)
Cytochrome P-450 Enzyme System/metabolism , Dantrolene/metabolism , Isoenzymes/metabolism , Microsomes, Liver/enzymology , Animals , Antibodies/pharmacology , Chromatography, High Pressure Liquid , Cytochrome P-450 Enzyme Inhibitors , Cytochrome P-450 Enzyme System/biosynthesis , Enzyme Induction , Female , Hydroxylation , Isoenzymes/antagonists & inhibitors , Kinetics , Male , Microsomes, Liver/metabolism , Rats , Rats, Inbred StrainsABSTRACT
1. Phencyclidine (PCP) was incubated with rabbit liver and brain microsomal fractions, and the structures of metabolites formed by oxidation determined by g.l.c.-mass spectrometry. 2. The formation of several known mono- and di-hydroxylated metabolites, as well as two new metabolites, was seen in the liver preparations. 3. Hydroxylated PCP metabolites were also formed after incubation of PCP with brain microsomes, indicating that PCP biotransformation may occur in the brain itself.
Subject(s)
Brain/ultrastructure , Microsomes, Liver/metabolism , Microsomes/metabolism , Phencyclidine/metabolism , Animals , Gas Chromatography-Mass Spectrometry , Hydroxylation , Male , RabbitsABSTRACT
The inductive potency of carboxylic acids, structurally related to clofibrate, on bilirubin UDPglucuronosyltransferase was investigated in the rat. For this purpose, structure-induction relationships were established using ten different arylcarboxylic or chlorophenoxycarboxylic acids. 4'-Chlorophenoxyacetic, -propionic and -isobutyric (clofibric) acids progressively increased hepatic glucuronidation of bilirubin (17%, 43%, 60% greater than controls, respectively) after a 5-day treatment in rat (100 mg/kg per day). 2-Phenylpropionic acid also enhanced bilirubin UDPglucuronosyltransferase activity (50%) in contrast to phenylacetic acid. The other compounds did not, or only slightly, affect this parameter. These results indicate that specific structural features are required for the induction property. Moreover, a good correlation (r = 0.962) was found between the extent of induction and the physiochemical descriptors which characterize the electronic state of the molecules, when analysed by multidimensional regression. Fluorescence polarization revealed that the compounds tested, especially clofibric acid, did not affect, in vivo or in vitro, the anisotropy of two different probes embedded in the microsomal membranes. Finally, since the interaction of the carboxylic acids with the membranes did not modify the latency state of bilirubin UDPglucuronosyltransferase, it was concluded that the increase in enzyme activity was due more to a real induction than to activation of bilirubin UDPglucuronosyltransferase. A close linkage was established between bilirubin UDPglucuronosyltransferase induction and that of cytochrome P-452, as shown by enhanced omega-oxidation of lauric acid. This led to the hypothesis that both processes could be under coordinate regulation and mediated by a molecular interaction depending on the physicochemical properties of the carboxylic acids.
Subject(s)
Bilirubin/metabolism , Clofibrate/analogs & derivatives , Clofibrate/pharmacology , Glucuronosyltransferase/metabolism , Lauric Acids/metabolism , Microsomes, Liver/metabolism , Animals , Cytochrome P-450 Enzyme System/metabolism , Enzyme Induction , Glucuronates/metabolism , Hydroxylation , Kinetics , Male , Rats , Rats, Inbred Strains , Structure-Activity Relationship , ThermodynamicsABSTRACT
Strict anaerobic cultures of Mucor hiemalis lead to major modifications of the quantitative and qualitative composition of sterols. Sterols of which the lateral chain contains an ethyl or ethylidene group in C24 are for the first time brought to evidence in a Mucorales. Isolation in the same conditions of E-ethylidene-24 (28) lanostene-8 ol-3 beta lets us suppose that sterols, which are absent in aerobic cultures, are derived from this triterpene of 32 C atoms.
