Biomedical subjects
Publications and source records attributed to C Riche.
The neuropsychiatric complications of histamine H2 receptor antagonists are poorly described in children. In this case report, lethargy occurred in a neonate treated with ranitidine. We urge caution when a histamine H2 receptor antagonist needs to be prescribed in such cases.
1. Interspecies (including man, monkey, rabbit, rat and mouse) variations in caffeine metabolism by liver microsomes were studied. While N-3 demethylation was the major pathway in man (81% of total dimethylxanthines), N-7 demethylation was predominant in monkey (89%), and the three demethylation pathways were about equal in mouse, rabbit and rat. 2. Three monooxygenase activities (methoxyresorufin O-demethylase, phenacetin O-deethylase and acetanilide 4-hydroxylase) correlated significantly with the rate of metabolism of caffeine. 3. P4501A1 and 1A2 enzymes were immunodetected in different species. P4501A2 was the only isoform detected in liver of man, rat and mouse, while no polypeptide immunorelated to P4501A was detected in monkey and only a minor band of P4501A1 was detected in rat and rabbit. 4. All in vitro data indicate that paraxanthine formation is mediated mainly by P4501A2 in mammals while theophylline formation is mediated mainly by cytochromes P-450 other than those of the 1A family.
The antiestrogen tamoxifen (Tam or Nolvadex, ICI)-Z-1-[4-[2-(dimethylamino) ethoxy]phenyl]-1,2-diphenyl-1-butene is widely used in treatment of hormone-dependent breast cancer. The drug is extensively metabolized by cytochrome P450 dependent hepatic mixed function oxidase in man, yielding mainly the N-desmethyl metabolite (DMT). This study has been carried out to determine the P450 enzyme involved in the N-oxidative demethylation of Tam in microsomal samples from 25 human livers (23 adults, two children). This metabolic step was inhibited by carbon monoxide up to 75%. Tam was demethylated into DMT with an apparent Km of 98 +/- 10 microM; rates varied between 37 and 446 pmol/min/mg microsomal protein. These metabolic rates were strongly correlated with 6 beta-hydroxylation of testosterone (r = 0.83) and erythromycin N-demethylase (r = 0.75), both activities known to be associated with P450 IIIA enzyme. To further assess whether or not the Tam demethylation pathway is catalysed by the same P450, the inhibitory effect of TST on this reaction was determined. The competitive inhibition had an apparent Ki of 100 +/- 10 microM. Drugs such as erythromycin, cyclosporin, nifedipine and diltiazem were shown to inhibit in vitro the metabolism of tamoxifen. Furthermore the P450 IIIA content of liver microsomal samples, measured by Western blot technique using a monoclonal P450NF (nifedipine) antibody, was strongly correlated with DMT formation (r = 0.87). Tam N-demethylase activity was inhibited by more than 65% with polyclonal anti-human anti-P450NF. All these in vitro observations establish that a P450 enzyme of the IIIA sub-family is involved in the oxidative demethylation of tamoxifen in human liver.
The metabolism of three anthracyclines, namely daunorubicin, doxorubicin and epirubicin, was studied in two human (HepG2 and HBG2) and one rat (FAO) hepatoma cell lines. Both species and substrate differences in anthracycline metabolism were retained by these hepatoma cell lines with regard to the aldoketoreductase pathway. However, glucuronidation of epirubicin normally found in vivo and in human hepatocyte cultures was not recovered in human hepatoma cells, further indicating that these cells express only some of the liver functions. Nevertheless, these cell lines could be a suitable model to investigate the still poorly understood aldoketoreductase activity.
Biotransformation of caffeine and theophylline and the effect of two well-known inducers of P-450 isozymes, namely phenobarbital (PB) and methylcholanthrene (3-MC) were studied in cultured hepatocytes from six human adult donors. Hepatocytes co-cultured with rat liver epithelial cells maintained a higher metabolic capacity than pure cultures. PB treatment of cultured hepatocytes for 3 days slightly increased the rate of caffeine metabolism 1.4 +/- 0.5-fold (N = 6) vs controls, and theophylline metabolism 1.2 +/- 0.4-fold (N = 6), whereas 3-MC treatment increased metabolism markedly 5.8 +/- 2.3- and 3.3 +/- 1.1-fold (N = 6) vs controls for caffeine and theophylline, respectively. Paraxanthine and theophylline formations from caffeine were the most induced by 3-MC. Their increase was significantly correlated (rs = 0.89, P less than 0.007) but not with TB formation, suggesting that at least two isozymes of the P-450IA family are involved in the first demethylations of caffeine. In addition, the N-1 demethylation of theophylline (mean increase of 554% vs controls) was not correlated with the N-1 demethylation of caffeine (mean to increase 247% vs controls) for the same donor after 3-MC treatment, suggesting that these two demethylations are mediated by a different P-450.
A method for extracting spiramycin by an octadecylsilica cartridge is described for plasma or vitreous samples. The macrolide antibiotic is then measured by reversed-phase HPLC with UV detection. The limit of detection is estimated to be 50 ng/mL. The coefficient of variation for the procedure is 6.1% and 5.2% for the range of concentrations 0.2 micrograms/mL and 10 micrograms/mL respectively. By this method, pharmacokinetic profiles were performed for five adult patients. Spiramycin could be accurately measured in the vitreous humour, allowing the determination of antibiotic at its site of action.
The pharmacokinetics of the tricyclic antidepressant amineptine (Survector) and its main metabolite were studied in 12 young healthy adults (6 men, 6 women; mean age 35.8 yr). Plasma samples were taken over 24 h following a single oral dose of 100 mg amineptine chloryhdrate. Plasma levels of both compounds were determined by means of high performance liquid chromatography. Amineptine was rapidly absorbed. Mean peak plasma concentrations of amineptine and its metabolite occurred 1 h and 1.5 h, respectively, after product administration. The mean apparent volume of distribution was large: 2.4 l.kg-1. Elimination was rapid; the mean half-lives of the 2 compounds were short: 0.8 h for amineptine and 2.5 h for the metabolite. The mean apparent plasma clearance of amineptine was high (124.8 l.h-1). When the results were adjusted for body weight and surface area, no significant difference in pharmacokinetic parameters was found between men and women. Given its pharmacokinetic characteristics there is no risk of amineptine accumulation and thus it is a particularly easy drug to manage. A standard dosage of amineptine was defined for use in healthy young adults.
1. Caffeine was used as a molecular probe for hepatic monooxygenase activity in three in vitro models from human livers: slices, microsomes and hepatocyte cultures. 2. A h.p.l.c. method for determination of all possible metabolites of caffeine (16 compounds) is described. 3. Caffeine biotransformation by these three in vitro systems was low. However, metabolite formation was shown to proceed at a rate close to that calculated from in vivo caffeine elimination half-life. 4. Metabolic profiles were the same whatever the in vitro system used. The ratio of primary demethylated metabolites was similar to that measured by in vivo studies, i.e. the selectivity for caffeine N-3 demethylation to paraxanthine was retained. 5. Significant correlation (p less than 0.001) between 7-ethoxyresorufin O-deethylation and caffeine demethylations was demonstrated for the 12 human livers studied.
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Cultured hepatocytes from newborn human (three samples), adult human (eight samples) and adult rat livers were used to study the metabolism of theophylline and caffeine, two drugs of which the metabolic pathways are known to be cytochrome P-450-dependent. Known metabolic pathways of caffeine in vivo were qualitatively maintained. However, only the primary metabolites were formed through oxidative N-demethylation giving theophylline, paraxanthine and theobromine and, through C-8 hydroxylation, giving 1,3,7-trimethyluric acid and a ring-opened compound the 6-amino-5[N-formylmethylamino]1,3-dimethyl uracil. The ratio of the three dimethylxanthine metabolites was dependent upon the species (human, rat), development stage (newborn, adult) and environmental factors. Similarly, theophylline was metabolized as in vivo by the demethylation pathway giving, preferentially, 3-methylxanthine and not 1-methylxanthine, and by a C-8 oxidation giving 1,3-dimethyluric acid. In newborn hepatocytes, all pathways were absent except the well-known methylation to caffeine. Moreover, such a methylation also occurred in adult human hepatocytes. This result was explained by the very low metabolic capacity of cultured cells, allowing the detection of only direct metabolites. Indeed, the overall biotransformation of both the methylxanthines by primary cultures of hepatocytes was remarkably weak, confirming previous studies with liver microsomal incubations. Thus the metabolism rate did not exceed about 30 nmoles/10(6) cells/24 hr in human adults, except for two subjects which were characterized by an extensive metabolism and a different metabolic profile. These two subjects were probably induced in vivo by environmental compounds. Both quantitative and qualitative data obtained from this study were roughly correlated with other in vivo and in vitro studies. Overall the experimental model of cultured human hepatocytes was shown to be capable of assessing the metabolic profile of two methylxanthines which is in agreement with the situation encountered in vivo. This example suggests that a breakthrough may be brought in new drugs development by the predictability from human hepatocyte culture model to the in vivo human situation.
An HPLC method was used to monitor the chemical stability of doxorubicin-HCl (DXR) to light in plasma, urine and cell culture medium at room temperature. The results indicated that DXR was very unstable in cell culture medium and urine when exposed to light. It was more stable in plasma under the same conditions. In all the cases, the decrease in the amount of DXR is greatly dependent on light intensity (no noticeable degradation was observed after 8 h in the dark). These observations may be important for the correct interpretation of the effects and the toxicity of doxorubicin on cells incubated in cell medium, and for determination of urinary or plasma pharmacokinetic parameters.
The X-ray crystal structure of 3-(methoxycarbonyl) amino-beta-carboline, a selective antagonist of the sedative effects of diazepam having a high affinity for the benzodiazepine receptor, has been determined. The results were compared with structural information obtained from this compound, both in the solid state and in dilute solution, by use of Fourier transform infrared spectroscopy. Its X-ray structure was also compared with those of two other active beta-carbolines, methyl beta-carboline-3-carboxylate and N-ethyl-3-carbamoyl-beta-carboline. The crystal packing characteristics of 3-(methoxycarbonyl) amino-beta-carboline differ from those of these two beta-carbolines in both the pattern of intermolecular hydrogen bonding and the quality of their pi-pi stacking interactions. The relevance this may have to the selective activity of 3-(methoxycarbonyl) amino-beta-carboline is discussed.
Maximal and minimal tobramycin concentrations were studied in 16 preterm and 4 fullterm neonates on the last day of a four-day course of tobramycin (infusion of 2 mg/kg twice a day). Maximal concentrations were under 6 mg/l in all patients and minimal concentrations were above 2 mg/l in one-third of the preterm babies. Next, tobramycin pharmacokinetic parameters were studied in 4 fullterm and 4 preterm neonates following a first infusion of 2 mg/kg. Half-life was 4 hours in term babies and 7.75 hours in preterm babies. Mean clearance was 0.118 l/h/kg and 0.06 l/h/kg in fullterm and preterm babies respectively (significant difference). Our results show that the 2 mg/kg dosage is too low and that, in preterm neonates, the 12-hour interval between infusions is too short.
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Phenobarbital was used as first treatment in 198 subjects with different loading doses adjusted according to age, followed by a daily maintenance dose of 5 mg/kg. A loading dose was given per os to 136 infants in four groups and intravenously to 62 subjects in five groups (including 30 adults). We found that the intravenous route must be used for the rapid production of therapeutic plasma concentrations and accordingly we recommend it in intensive care. Posology of 20 mg/kg i.v. appears to be adequate in children up to 15 years old while a posology of 15 mg/kg seems to be too high in adults. The maximum probability of efficiency is delayed up to 12 h when phenobarbital is given per os. Various posologies were found to be adequate in infants except for premature newborns.
25-hydroxyvitamin D (25 OH vit D) plasma levels were measured according to the method of Preece et al. in 2 groups of subjects. Group 1 consisted of 10 adult controls in whom 25 OH vit D levels were measured 11 times at 1 month intervals. Group 2 consisted of 93 mother-child pairs including 71 neonates with normal plasma calcium levels (group 2a) and 22 with hypocalcemia (group 2b). In mothers, 25 OH vit D levels (group 2a: 12 ng/ml +/- 6.4 SD, n = 71; group 2b: 9.5 ng/ml +/- 3.5 SD, n = 22) were lower than in controls (25.1 ng/ml +/- 7.9 SD). In neonates, 25 OH vit D levels were correlated with those of their mothers; however there was no significant difference in neonates with or without hypocalcemia (group 2a: means = 11.9 ng/ml +/- 6.1 SD, n = 71; group 2b: means = 10 ng/ml +/- 4.1 SD, n = 22). 25 OH vit D levels in adults of group 1 were significantly correlated with exposure to sunlight, while no significant correlation was found in the mothers-children pairs of group 2. These results confirm the relationship of 25 OH vit D plasma levels of neonates with their mothers' but do not support the concept of a relation between low plasma levels in mother and the occurrence of hypocalcemia in neonates.
Neonates with severe cerebral damage are usually given phenobarbital. The authors study the various means of administration and the influence of term and concomitant mannitol administration on the plasma levels of the drug. They recommend the intravenous administration of a load dose and show that phenobarbital blood levels are significantly less in prematures or in children who have been given mannitol.
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