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Biomedical subjects

D V Parke

Publications and source records attributed to D V Parke.

At least 91 records · Page 5Linked to original sources
PubMed1987

Interactions of imidazole antifungal agents with purified cytochrome P-450 proteins.

The imidazole N-substituted antifungal agents ketoconazole, miconazole and clotrimazole have been shown to be potent inhibitors of oxidative metabolism by both a phenobarbital-induced cytochrome P-450 (P-450b) and a 3-methylcholanthrene-induced cytochrome P-448-protein (P-450c) in reconstituted systems. All three compounds inhibited the cytochrome P-450b-dependent 7-pentoxyresorufin-O-dealkylase and the cytochrome P-450c-dependent 7-ethoxyresorufin-O-deethylase activities. When 7-benzyloxyresorufin and 7-ethoxycoumarin were employed as substrates with both cytochrome preparations, all three antifungal compounds exhibited selective inhibition of the cytochrome P-450b preparation; ketoconazole was always the weakest inhibitor. The three antifungal agents were also shown to elicit a type II difference spectral interaction with both isoenzymes, the magnitude of the spectral interaction being greater with the cytochrome P-450b preparation.

7-Alkoxycoumarin O-Dealkylase↗
PubMed1987

Activation mechanisms to chemical toxicity.

The pathobiology of chemical toxicity may involve "acute lethal injury" (necrosis), "autoxidative injury" (oxygen toxicity), "immunological injury" (neoantigen formation), and malignancy. Toxic chemicals may be activated by reduction, conjugation, radical formation, or oxidation. Oxidative activation may be effected by cytochromes P-450/P-448, flavoprotein monooxygenases, or hydroxyl radicals. The alternative pathways of oxidative metabolism of toxic chemicals, namely, detoxication and activation, are catalysed by the phenobarbital-induced cytochromes P-450 and by the 3-methylcholanthrene-induced cytochromes P-448 respectively. Oxidative metabolism by cytochromes P-450 is followed by conjugation and detoxication, whereas oxidative metabolism by cytochromes P-448 yields reactive intermediates which are not readily conjugated, and thus react with vital intracellular macromolecules, resulting in necrosis, redox cycling and oxygen radical formation, neoantigen production, and mutations. The molecular dimensions of specific substrates, inhibitors and inducers of the PB-cytochromes P-450 indicate that they are globular and are different from those of the cytochromes P-448 which are planar, suggesting that the active sites of the two families of enzymes are different. Oxidative metabolism of planar substrates of cytochromes P-448 results in conformationally-hindered oxygenations, which inhibits subsequent conjugations. Cytochrome P-448 activity may be quantified by the oxidative deethylation of 7-ethoxyresorufin which, unlike benzo(a)pyrene hydroxylation (AHH) is a specific reaction for this family of enzymes. Oxidative metabolism of chemicals varies inversely with the body weight of the animal species, so that chemical toxicity involving oxidative activation, redox cycling, and reactive oxygen is greater the smaller the animal species.

Animals↗
PubMed1987

Structural requirements for substrates of cytochromes P-450 and P-448.

Distinct and different molecular structural features are manifested by substrates, inhibitors and inducers of the two families of liver microsomal enzymes, the phenobarbital-induced cytochromes P-450 and the 3-methylcholanthrene-induced cytochromes P-448. In a theoretical study based on molecular orbital calculations and molecular graphics, it is established that cytochrome P-448 substrates contain fused aromatic or heteroaromatic rings giving rise to overall molecular planarity with relatively small molecular depth. In contrast, substrates of the cytochromes P-450 have greater conformational freedom and an ability to bind at more than one point of attachment, as a result of possession of certain characteristic functions, namely, a carbonyl and/or amine moiety coupled with an iso-propyl group, or similar function of equivalent shape and hydrophobicity. The implications are that the binding sites of cytochromes P-448 contain a number of hydrophobic aromatic amino acid residues orientated so as to allow occupation by similar substrates containing co-planar aromatic rings, whereas those of the phenobarbital-induced cytochromes P-450 contain hydrophilic amino acid residues capable of hydrogen bonding to greater than C = O moieties and at least one leucine or valine residue, as these contain the complementary isopropyl function. The corollary of these findings is the possibility of prediction of the toxicity of new chemicals on the basis of their molecular dimensions.

Binding Sites↗
PubMed1987

Spectral and kinetic studies of the interaction of imidazole anti-fungal agents with microsomal cytochromes P-450.

1. The imidazole antifungal agents, ketoconazole, miconazole and clotrimazole have been shown to be potent inhibitors of the phenobarbital-induced cytochromes P-450 and the 3-methylcholanthrene-induced cytochromes P-448-dependent rat hepatic microsomal mixed-function oxidases. 2. All three drugs were more potent inhibitors of the phenobarbital-induced O-deethylation of ethoxycoumarin than of the 3-methylcholanthrene-induced activity indicating selective inhibition of the phenobarbital-induced cytochromes P-450. In both types of microsomes ketoconazole was always the weakest inhibitor. 3. All three compounds elicited type II spectral interactions with both types of microsomes, and had similar Ks values. Miconazole and clotrimazole, and to a lesser extent ketoconazole, also interacted with the substrate binding sites of both phenobarbital-induced cytochromes P-450 and to a lesser extent with the 3-methylcholanthrene-induced cytochrome P-448. 4. It is concluded that at least part of the inhibitory effect of these antifungal agents may reflect competitive inhibition at the substrate binding site.

Animals↗
PubMed1986

Induction of cytochrome P-448 activity as exemplified by the O-deethylation of ethoxyresorufin. Effects of dose, sex, tissue and animal species.

The effects of tissue, sex, animal species and dose on the induction of cytochrome P-448 activity by various inducing agents were investigated using O-ethoxyresorufin as a model substrate. The liver was by far more effective in catalysing the O-deethylation of ethoxyresorufin (EROD) than the lung and kidney. The extent of induction was also highest in the liver, with the exception of benzo(a)pyrene and 3-methylcholanthrene where inducibility was more pronounced in the kidney. The benzo(a)pyrene-induced hepatic EROD activity in the rat decayed to reach control levels four days after a single administration. Rat hepatic EROD activity was induced in both sexes but tended to be higher in the male. Marked species differences in the inducibility of hepatic EROD activity by various chemicals was observed, the rat being always more responsive when compared to the hamster or mouse. The induction of rat hepatic EROD activity by benzo(a)pyrene, 2-acetylaminofluorene and safrole was dose-dependent, maximum induction being achieved with single doses of 5, 2 and 5 mg/kg, respectively.

Animals↗
PubMed1986

Molecular dimensions of the substrate binding site of cytochrome P-448.

The molecular geometries of specific substrates, inhibitors and inducers of cytochrome P-448 activity were determined using computer-graphic techniques for use in defining the molecular dimensions of the substrate binding site of this enzyme. Specific substrates of cytochrome P-448 are essentially planar molecules characterised by a small depth and a large area/depth ratio. In contrast, compounds that do not serve as substrates of cytochrome P-448 are bulky, non-planar molecules characterised by small area/depth ratios and greater flexibility in molecular conformation. Specific inhibitors of cytochrome P-448 whose effect is mediated through interaction with the haem still meet the dimensional criteria for substrates indicating that they must also interact with the substrate binding-site, which is probably located in proximity to the haem. Inducers of cytochrome P-448 activity exhibit similar molecular geometries to the substrates from which it may be inferred that the cytosolic receptor associated with the induction of cytochrome P-448 activity is structurally related to the active site of the cytochrome.

Animals↗
PubMed1986

Free radical production at the site of an acute inflammatory reaction as measured by chemiluminescence.

A foot-pad oedema model was used to investigate the presence of free radicals using a chemiluminescence method. This model is an example of a cell mediated hypersensitivity reaction. Male rats were inoculated in the scruff with Freund's Complete Adjuvant (FCA) on Day 0 and then challenged 6 days later with FCA in one hind paw. An acute inflammatory reaction was initiated over the following 96 hours and within 4 hours of induction, reactive oxygen species were detected in the inflamed tissue. A peak of chemiluminescence activity was seen 8 hours after the induction of the inflammatory reaction, well before maximum oedema was observed. Using mannitol, catalase and DABCO to elucidate the nature of the reactive oxygen species it was found that hydroxyl radicals, hydrogen peroxide and singlet oxygen all contributed to this burst of oxidative activity and are therefore probably involved with the process of lipid peroxidation and the severity of an inflammatory reaction.

Animals↗
PubMed1986

Liver function in patients on long-term paracetamol (co-proxamol) analgesia.

Eleven patients on long term co-proxamol therapy for pain, in general practice, showed no abnormalities of liver or renal function, as assessed by serum prealbumin concentrations and blood enzyme and electrolyte activities. This indicates that although the preparation is hepatotoxic when taken acutely in overdose, in chronic therapeutic dosage it appears to be free from this hazard.

Acetaminophen↗
PubMed1986

Further studies on the pharmacokinetics of perhexiline maleate in humans.

We have performed single-dose pharmacokinetic studies on perhexiline in eight young volunteers, each given 300 mg of Pexid orally, using an h.p.l.c. method for the separation and quantification of the drug and its monohydroxy metabolites in plasma and urine. The plasma concentration of the cis-monohydroxyperhexiline (peak of 473 +/- 43 ng/ml at 7.5 +/- 2.0 h) was always higher than for unchanged perhexiline (peak of 112 +/- 20 ng/ml at 6.5 +/- 2.0 h) whereas the concentration of the transmetabolite was either low or undetectable in plasma. These findings indicate the occurrence of stereospecific pre-systemic metabolism of perhexiline which reduces the bioavailability of the parent drug. The plasma elimination half-life of perhexiline was 12.4 +/- 6.1 h (range 7-23 h) while that for cis-monohydroxyperhexiline was 19.9 +/- 7.7 h (range 10-29 h). Not more than 0.3% of unchanged perhexiline was excreted in the urine over five days in eight subjects. Between 3 and 23% of the orally administered drug was excreted as the cis- or trans-monohydroxy metabolites, the ratio of trans to cis metabolites being 0.52 +/- 0.20.

Adult↗
PubMed1986

Effects of brotizolam on mixed-function oxidases and glutathione metabolism in the rat.

Intra-gastric administration of brotizolam (0.1-200 mg/kg) daily for three days to rats resulted in no significant changes in the hepatic and intestinal cytochrome P-450-dependent or P-448-dependent mixed-function oxidases, or in the hepatic flavoprotein dimethylaniline N-oxidase. Liver microsomes from mouse, rat and man metabolized brotizolam by hydroxylation of the diazepine ring and of the methyl group at rates which were greater for mouse greater than rat greater than man. Brotizolam and its metabolites generated by rat-liver microsomes in vitro were not mutagenic in the Ames' test. Brotizolam, at 200 mg/kg per day for two to six weeks, depleted liver glutathione concentration and markedly increased liver gamma-glutamyl transpeptidase, glutathione reductase and glutathione transferase activities. Similar changes were not seen at the lower dose of 0.3 mg/kg. The observed increases in glutathione metabolism and the decreased tissue concentration of glutathione are indicative of high levels of glutathione conjugation, and provide a possible explanation for the equivocal increase in tumorigenicity seen in rats receiving brotizolam at high dosage.

Animals↗
PubMed1986

Stereoselective pharmacokinetics of perhexiline.

Blood plasma and urine excretion pharmacokinetics of the (+) and (-) enantiomers of perhexiline have been determined in oral single-dose studies in eight human volunteers, and compared with the pharmacokinetics of the racemate drug in the same subjects. The (-) enantiomer is more rapidly metabolized and eliminated, and is stereoselectively hydroxylated to the cis-monohydroxy-perhexiline. The peak plasma concn of unchanged perhexiline is greater, while that of the cis-monohydroxy-perhexiline metabolite is lower, after administration of the (+) enantiomer than after the (-) enantiomer or the racemate. Similarly, the AUC values for unchanged perhexiline and for the trans-monohydroxy-perhexiline metabolite are greatest and the AUC value for the cis-monohydroxy-perhexiline metabolite is lowest for the (+) enantiomer. The three stereoisomeric forms of perhexiline all had the same times to peak plasma concn of the unchanged drug or of the cis-metabolite, and all three forms had a similar plasma elimination half-life for unchanged perhexiline. Metabolism of racemic perhexiline to the cis-monohydroxy metabolite is the major mechanism of elimination of the drug in man and has been shown to be polymorphic in human populations. The (-) enantiomer which shows stereoselective metabolism to the cis metabolite might therefore show a greater polymorphic effect. Studies with rat-liver microsomal preparations in vitro showed that, in contrast to the human studies in vivo, hydroxylation of perhexiline yields mostly the trans-monohydroxy metabolite. The DA strain of rats exhibited slower rates of hydroxylation in vitro than Wistar or Lewis strains of rats.

Adult↗
PubMed1985

The homogeneity of rat liver microsomal cytochrome P-448 activity and its role in the activation of benzo[a]pyrene to mutagens.

The O-deethylation of ethoxyresorufin and the metabolic activation of benzo[a]pyrene to mutagens were determined in hepatic microsomal preparations from control and induced animals. An excellent direct correlation (r = 0.95) has been observed between ethoxyresorufin O-deethylase and the metabolic activation of benzo[a]pyrene to mutagens when the fraction of cytochromes P-450 present as cytochrome P-448 was altered by the administration of phenobarbitone and 3-methylcholanthrene alone or in combination with 9-hydroxyellipticine. The correlation between these activities was maintained following treatment of animals with Arochlor 1254, benzo[a]pyrene, benzo[e]pyrene, 7,12-dimethylbenzo[a]anthracene,2-anthramine and 2-naphthylamine.

Animals↗
PubMed1985

An assessment of a toxicological incident in a drug development program and its implications.

In an exceptionally thorough program of preclinical toxicity testing in laboratory animals of the anti-allergy drug FPL 52757 (6,8-diethyl-5-hydroxy-4-oxo-4H-1-benzopyran-2-carboxylic acid), no significant hepatotoxic potential was detected. Despite this evidence of safety in a wide range of laboratory animals, the drug produced mild reversible hepatotoxicity in some patients in some clinical trials. Extensive toxicity and investigational studies are described, and the importance of the correct choice of species for toxicology evaluations based on sound pharmacokinetic principles is highlighted. This paper reviews this incident and the lessons to be learned from it in light of current regulatory requirements.

Animals↗
PubMed1985

Interactions of safrole and isosafrole and their metabolites with cytochromes P-450.

The structural features which determine interaction of safrole and related methylenedioxyphenyl compounds with cytochromes P-450 or P-448, and determine the induction of these two classes of the cytochrome, have been studied. All methylenedioxyphenyl compounds studied interact with both cytochromes P-450 and P-448 eliciting type I spectral changes and it has been found that the allyl 4-substituent is important in these interactions. Methylenedioxyphenyl compounds with an oxidised allyl 4-substituent exhibited higher affinity for cytochrome P-448 while those possessing an intact allyl or methylvinyl group generally showed higher affinity for cytochrome P-450. Compounds possessing intact allyl and methylenedioxyphenyl groups (safrole, isosafrole and myristicine) were the most potent inducers of cytochromes P-450 and P-448; compounds containing an intact allyl group only (estragole, allybenzene and eugenol methyl ether) or an oxidized allyl group and an intact methylenedioxyphenyl group (epoxysafrole) were inducers of P-448 only.

Animals↗