Disposition of gabapentin (neurontin) in mice, rats, dogs, and monkeys.

Gabapentin, an analog of gamma-aminobutyric acid, exhibits anticonvulsant properties in both animal models and humans. Gabapentin pharmacokinetics was studied in laboratory animals using HPLC and radiometry. Oral bioavailability was 40% in monkeys administered 25 mg/kg, 79% in mice and rats receiving 50 mg/kg, and 80% in dogs administered 50 mg/kg. Binding to plasma proteins was < 3%. Maximum blood or plasma concentrations generally occurred within 2 hr of an oral dose. In rats and monkeys, increases in maximum plasma concentrations and/or areas under the curve were less than dose-proportional following oral administration, most likely because of saturable absorption. However, intravenous pharmacokinetics in rats were linear over the dosage range of 4-500 mg/kg. Mean intravenous elimination half-life was 1.7 hr in rats, 2.9 hr (14C only) in dogs, and 3.0 hr in monkeys. In rats and dogs, repeated administration did not alter gabapentin or 14C pharmacokinetics. Additionally, gabapentin did not induce hepatic cytochrome P450 monooxygenases in rats. There were no age- (rats only) or gender-associated changes in pharmacokinetic parameters. [14C]Gabapentin was extensively distributed to tissues. In the dog, gabapentin was metabolized to N-methylgabapentin (approximately 34% of dose); whereas metabolism in mouse, rat, and monkey was minimal (< 5%). The principal route of excretion was via urine. In summary, as an antiepileptic drug, gabapentin exhibited desirable pharmacokinetic properties, such as linear elimination kinetics, not highly bound to plasma proteins, not extensively metabolized, and not an inducer of hepatic cytochrome P450.

Pharmacokinetics of tilidine and metabolites in man.

Tilidine is a prodrug from which the active metabolite nortilidine is formed by demethylation. The pharmacokinetics of tilidine (T), nortilidine (NT) and bisnortilidine (BNT) were studied in nine healthy subjects following single intravenous (10 min infusion) and oral 50 mg T-HCl dose as well as following multiple 50 mg T-HCl oral doses. Systemic availability of the parent substance was 6% and of the active metabolite NT 99%. The terminal half-life of NT was 3.3 h following single oral administration, 4.9 h following intravenous administration and 3.6 h following multiple dosing. Following intravenous infusion, concentrations of unchanged substance were found which were 30 times higher than following oral administration. BNT was eliminated with half-lives of 5 h after oral administration and 6.9 h after intravenous administration. Renal elimination of unchanged substance was 1.6% of the dose following intravenous administration and less than 0.1% of the dose following oral administration. Approximately 3% were recovered in urine as NT and 5% as BNT following both routes of administration.

Pharmacokinetics and metabolism of gabapentin in rat, dog and man.

This paper describes the pharmacokinetic studies of 1-(aminomethyl)-cyclohexane acetic acid (gabapentin, Gö 3450, CI-945) conducted with the 14C-labelled substance following intravenous and intragastric administration to rats and dogs and oral administration to humans. Gabapentin is well absorbed in rats, dogs and in humans, with maximum blood levels, reached within 1-3 h after peroral administration. Following i.v. administration to rats, similar blood and brain levels of gabapentin are observed after a short distribution phase, whereby concentrations in cerebrum and cerebellum are comparable. The highest concentrations are found in the pancreas and kidneys and the lowest values in adipose tissue. No binding of gabapentin to human plasma proteins or human serum albumin is observed. The distribution coefficient (octanol/buffer pH 7.4) is 7.5 X 10(-2). In man, no biotransformation of gabapentin is observed. In rats, biotransformation is only minor. In dogs, however, a remarkable formation of N-methyl-gabapentin is found. Elimination half-lives range between 2-3 h in rats, 3-4 h in dogs, and 5-6 h in man. Gabapentin is nearly exclusively eliminated via the kidneys. Renal elimination was up to 99.8% in rats and approx. 80% in man following oral administration. The blood level-time course after i.v. administration to rats can well be described by a three-compartment open model. Experiments in rats and dogs demonstrate that pharmacokinetics are not sex-dependent and are not changed after multiple dosage. Pharmacokinetics are shown to be linear in the range tested of 4 to 500 mg/kg i.v. in rats.

High performance liquid chromatography coupled with radioactivity detection: a powerful tool for determining drug metabolite profiles in biological fluids.

High performance liquid chromatography coupled with continuous radioactivity detection represents an advancement in drug metabolism research. Using radioactive substances labelled in biologically stable positions, all metabolites can be specifically detected by radioactivity measurement. Thus no clean-up of biological fluids is required prior to HPLC. This can prevent artefact formation from unstable metabolites, reduces recovery problems and facilitates quantitation. Separation of highly polar and unpolar metabolites is possible in a single chromatographic run using gradient elution and reversed phase materials. This technique is also well-suited for preparative isolation and purification of metabolites for subsequent structure elucidation. Various metabolite profiles of drugs labelled with carbon-14 or tritium are shown. Metabolites of the following drugs are presented: norfenefrine, etozolin, thymoxamine, naloxone, and levobunolol. We review the general methodology and report our experience with this technique. In principle, this technique may be useful for all biological systems in which tracer techniques are applied.

Pharmacokinetics of isoxicam following intravenous, intramuscular, oral and rectal administration in healthy volunteers.

1 The pharmacokinetic characteristics and bioavailability of isoxicam were investigated in healthy male volunteers following various routes of administration. Plasma concentrations were determined up to 96 or 120 h following each administration using a selective h.p.l.c. method. 2 Twelve subjects received an i.v. and an i.m. injection of 150 mg and an oral dose of 200 mg (two capsules) in a three-way crossover design. 3 The plasma concentration-time curves after intravenous administration followed two compartmental characteristics with a rapid initial distribution phase and a predominant terminal elimination phase. The area under the distribution phase contributed only 0.4% to the total AUC. Mean disposition parameters were: t1/2, lambda 1 = 3.3 min t1/2, lambda z = 28 h, Vc = 5.61, Vdarea = 12.71, CL = 5.1 ml min-1. 4 Following other routes of administration, the isoxicam plasma profile could be adequately described by a one compartment model. 5 After the i.m. dose, isoxicam was rapidly and completely absorbed: within 15 min 40% of the peak concentrations were attained. Maximum plasma concentrations of 11.7 micrograms ml-1 were reached after an average of 3 h. 6 After oral administration isoxicam was completely bioavailable. Mean peak concentrations of 12.3 micrograms ml-1 were obtained within 10 h. 7 The bioavailability of suppositories containing 200 mg of active substance was investigated in another 10 volunteers relative to capsules (2 X 100 mg). The formulations tested were bioequivalent with respect to the amount absorbed. 8 Comparison of AUCs (normalized for dose and body weight) resulting from all treatments, did not show significant differences among the various formulations.(ABSTRACT TRUNCATED AT 250 WORDS)

GLC method for the quantification of metabolites of thymoxamine in human plasma.

A sensitive gas-chromatographic method for quantification of the pharmacologically active metabolites I-IV of thymoxamine in plasma is described. 4-(Hydroxythymyl)-(2'methylbutylaminoethyl)ether, a compound similar to metabolite I, is used as an internal standard. Metabolites I and II the internal standard are extracted with cyclohexane from alkalinized plasma followed by back-extraction into 0.1 N hydrochloric acid. After evaporating the hydrochloric acid solution, the sample is silylated with BSTFA and analyzed by gas-chromatography on a CRS 101/Carbowax 4000 column using a thermoionic detector. For subsequent determination of metabolites III and IV, the extracted plasma is hydrolyzed under conditions in which the phenol sulfates but not the glucuronide conjugates undergo cleavage. The resulting phenols (metabolite I and II) are analyzed as described above. The sensitivity threshold for all 4 compounds is approximately 5 ng/ml plasma based on a 2 ml plasma sample.

Metabolism of thymoxamine. III. Structure elucidation of the metabolites and interspecies comparison.

The structures of six metabolites were elucidated using rat urine after intragastric administration of 14C-thymoxamine by means of enzyme incubations, mass spectrometry and synthesis of metabolites: desacetylthymoxamine, N-demethyl-desacetylthymoxamine, the corresponding sulfates and glucuronides. The nature of the conjugates was confirmed by biosynthesis, i.e., co-administration of unlabelled thymoxamine and 35S-sulfate or 14C-glucose. The system high performance liquid chromatography-radioactivity detection was used for interspecies comparison. All biotransformation pathways are seen in rat and man. In dog and cat demethylation is a very minor reaction. Glucuronidation is not observed in the cat.

Metabolism of thymoxamine. II. Studies with 14C-thymoxamine in man.

Thymoxamine is rapidly and completely absorbed in man. Rapid biotransformation is observed after intravenous and oral administration of 40 mg 14C-thymoxamine HCl. No unchanged compound is found in the body. More than 90% of plasma and urine radioactivity could be ascribed to six metabolites: the desacetyl compound (metabolite I), the monodemethylated metabolite I (metabolite II), the sulfate conjugates of I and II (metabolites III and IV) and the glucuronides of I and II (metabolites V and VI). The unconjugated metabolites are observed in plasma only after intravenous administration. Similar patterns for polar metabolites are found in plasma and urine for both routes of administration. The sulfate fraction amounts to about 50-60% and the glucuronide fraction to about 30-40% of the radioactivity, the conjugates of metabolite I being more abundant than those of metabolite II. The elimination of the metabolites is rapid, the half-life of radioactivity elimination being 1.5 h during the first 12 hours and 12 h thereafter. 80% of the radioactivity dose is recovered in the urine within 4 hours. Recovery after four days amounts to 99.8% (i.v.) and 97.7% (oral). The results are discussed with regard to the application of the drug in man, taking into account that not only the unconjugated metabolites but also the sulfate conjugates are pharmacologically active.

Metabolism of thymoxamine. I. Studies with 14C-thymoxamine in rats.

Thymoxamine is rapidly and completely absorbed in rats. It is a prodrug which does not enter the systemic circulation in its unchanged form. After either oral or intravenous administration it undergoes rapid and intense metabolism involving four biotransformation reactions: Enzymatic hydrolysis to the corresponding phenol (metabolite I), Monodemethylation to metabolite II, Sulfate conjugation of I and II (metabolites III and IV) and Conjugation of I and II with glucuronic acid (metabolites V and VI). With these 6 metabolites identified approximately 95% of the radioactivity can be accounted for in plasma, urine and bile. Whereas the systemic availability of I and II is low, III and IV show high bioavailability. Metabolites I to IV are pharmacologically active, while III and IV are less potent than I and II. The radioactivity distribution in tissues is different after oral and intravenous administration consistent with the higher portion of unconjugated metabolites in the body after administration by parenteral route. Although 60% of the labelled compounds is eliminated via bile, the radioactive compounds are almost completely excreted in the urine after both routes of administration. This demonstrates complete reabsorption of the biliary metabolites. Secondary peaks of radioactivity in plasma and organs at 4 hours are explained by the participation of the metabolites in the enterohepatic circulation.

Binding of beta-adrenoceptor antagonists to rat and rabbit lung: special reference to levobunolol.

Binding of 3H-dihydroalprenolol (3H-DHA) to beta-adrenoceptors in homogenates from rat and rabbit lung was homogeneous and of high affinity (KD = 0.6 and 1.1 nmol/l at 20 degrees C; 1.1 and 2.2 nmol/l at 37 degrees C). The beta 1-selective antagonists betaxolol, metoprolol, bevantolol and acebutolol displaced 3H-DHA in a biphasic manner. From these data, the beta-adrenoceptor subtype distribution in rat lung homogenates was estimated to be 80% beta 2 (at 20 and 37 degrees C) as compared to 25% beta 2 in rabbit lung homogenates. In general, binding of beta-adrenoceptor antagonists (selective and nonselective) was slightly (less than 2 X) weaker in rabbit than in rat lung homogenates. In rat lung, binding of cardioselective beta-blockers to beta 1-receptors seemed to be more temperature-sensitive than binding to beta 2-receptors or binding of nonselective beta-blockers. Levobunolol, a potent non-cardioselective beta-blocker in pharmacological experiments, displaced 3H-DHA in a homogeneous manner (indicative of non-selectivity). In rat lung homogenates KD values were 0.8 nmol/l at 20 degrees C and 2.1 nmol/l at 37 degrees C. Similar values were found for the metabolites dihydrolevobunolol and hydroxylevobunolol. Surprisingly, d-bunolol, the dextrarotatory enantiomer of bunolol, showed a biphasic displacement curve, the fraction of high affinity sites being 83% in rat lung homogenates and 23% in rabbit lung. This ratio of sites is expected for a beta 2-adrenoceptor preferring ligand. High affinity binding (i.e. supposedly binding to beta 2-receptors) was about 50 times weaker than binding of levobunolol, in agreement with known stereospecificity of beta-adrenoceptor binding.

Pharmacokinetic model of diltiazem.

The pharmacokinetic profile of D-3-acetoxy-cis-2,3-dihydro-5-(2-dimethylamino-ethyl)-2-(p-methoxy-phenyl)-1, 5-benzothiazepin-4(5H)-one hydrochloride (diltiazem . HCl) following i.v. and p.o. administration has been studied in six healthy subjects using a new sensitive GLC method. The volunteers received an i.v. infusion of 20 mg in 20 min, a peroral solution of 120 mg and two 60-mg tablets (Dilzem) in a randomized sequence. The plasma level time courses of the unchanged compound following infusion and peroral solution were simultaneously evaluated by nonlinear regression analysis. The best model was chosen by means of statistical criteria. In all subjects the experimental data could be adequately described by an open three-compartment model with zero order input following infusion and first order absorption following p.o. solution. Using this procedure the following common disposition parameters were obtained for both routes of administration: t1/2 alpha = 0.1 h, t1/2 beta = 2.1 h, t1/2 gamma = 9.8 h (harmonic means derived from individualized fits), Vc = 0.9 +/- 0.4 l/kg, Vss = 5.2 +/- 2.4 l/kg, Cltot = 11.5 +/- 1.8 ml/min/kg. Compared to the beta-phase the terminal gamma phase represents a smaller contribution to the total AUC. The blood/plasma distribution ratio was found to be 1.00 +/- 0.08 (N = 4), the mean hepatic extraction ratio was 0.54. Peak levels appeared 0.6 +/- 0.3 h after the p.o. solution. The mean absolute bioavailability of diltazem based on the individual AUC infinity 0 of the p.o. solution and the infusion was 0.44 +/- 0.10. Following tablet administration, delayed maximum levels were found after 2.8 +/- 0.9 h. Comparing the AUC infinity 0 of tablets and p.o. solution, there was no significant difference between both dosage forms.

Temperature dependence of the benzodiazepine-receptor interaction.

The temperature dependence of the interaction of three benzodiazepines with their receptors in rat brain membranes containing about 2 microM (endogenous) gamma-aminobutyric acid was investigated. van't Hoff plots of the equilibrium dissociation constants were linear for [3H]diazepam and its N-desmethyl derivative (N-desmethyldiazepam, NDz), whereas for [3H]flunitrazepam a break between two linear portions occurred at about 10 degrees. Binding of diazepam, NDz, and flunitrazepam (for the latter, at temperature greater than 10 degrees) is enthalpy-driven (delta H approximately -40 to -50 kJ/mole with a negligible contribution from delta S. The latter result indicates that the interaction is not a simple hydrophobic association, but that it may be more complex in nature, possibly involving a conformational transition of the receptor-ligand complex. The activation energy for dissociation of the [3H]flunitrazepam- and [3H]diazepam-receptor complexes is about 100 kJ/mole. Consequently, the complexes dissociate about 100 times faster on changing temperature from 0 degrees to 37 degrees.

Assay of etozolin and its main metabolite, ozolinone, in plasma by high performance liquid chromatography.

A sensitive and specific method for the determination of the diuretic Ethyl (Z)-(3-methyl-4-oxo-5-piperidino-thiazolidin-2-ylidene) acetate (etozolin, Elkapin) and its pharmacologically active main metabolite, (Z)-(3-methyl-4-oxo-5-piperidino-thiazolidin-2-ylidene) acetic acid (ozolinone), at therapeutic concentrations in plasma is described. The method is based on high performance liquid chromatography and the use of two structurally similar internal standards. Etozolin and its metabolite are extracted from the plasma into dichloromethane at pH 9 and pH 5, respectively, and the resulting residues are analyzed on a silica gel column. The elution peaks are detected by UV absorption at 281 nm. The sensitivity for both compounds is 20 ng/ml plasma. The precision of the method is about +/- 5%. The applicability to pharmacokinetic studies of etozolin is shown.

GLC determination of tilidine, nortilidine, and bisnortilidine in biological fluids with a nitrogen-sensitive detector.

A sensitive and specific GLC method is described to determine therapeutic levels of tilidine and its two main metabolites, nortilidine and bisnortilidine, in plasma and urine. The method involves the extraction of the compounds and an internal standard with cyclohexane from alkalinized samples, followed by back-extraction into 1 N HCl. The hydrochloric acid solution is evaporated to dryness. After liberation of the free bases with ammonia, the residue is subjected to GLC analysis with a nitrogen-phosphorus detector and a 1.8-m (6-ft) glass column packed with 1% CRS 101 and 1.5% LAC-4-R-886 on Gas Chrom Q. Sensitivity in plasma and urine is approximately 1 ng/ml for a 5-ml sample.

Gas-chromatographic determination of nanogram amounts of enantiomers of nortilidine, a main metabolite of tilidine, in biological specimens.

We report a specific and sensitive method for determination of the individual optical isomers of nortilidine, a main metabolite of tilidine, with the aid of a nitrogen-sensitive detector. With N-trifluoroacetyl-L-leucyl chloride as chiral reagent, the diastereomeric derivatives of the nortilidine enantiomers could be separated and quantified in the nanogram range. Under these conditions, the enantiomers of bisnortilidine, another main metabolite of tilidine, were also separated. Investigations in rats with the enantiomers of tilidine and nortilidine indicated that no racemization occurs during N-demethylation in the organism. After oral and intravenous administration of 50 mg of tilidine.HCI to a human volunteer, identical concentrations of nortilidine enantiomers were found in the plasma.

[Metabolism of Etozolin in rat, dog and man]. / Metabolismus von Etozolin bei Ratte, Hund und Mensch.

Investigations on the metabolism of the new diuretic ethyl (Z)-(3-methyl-4-oxo-5-piperidino-thiazolidin-2-ylidene) acetate (Gö 687, etozolin, Elkapin) were carried out with urine of rat, dog and man as well as rat bile after enteral administration of the 14C-labelled substance. Seven metabolites were isolated with either the aid of high-pressure liquid chromatography (HPLC) or extraction and thin-layer chromatography. Mass spectroscopy was applied to determine the structures of the metabolites, partly by use of authentic reference substances. Because of the instability of most of the metabolites, some of them showing strong polarity, the described investigations on the metabolic profiles and the enrichment and purification of some metabolites could only be carried out with the HPLC-radioactivity detector system, which requires no clean-up for the samples. The metabolisation process of etozolin is qualitatively equal in rat, dog and man; it is characterized by 3 steps: 1. enzymatic cleavage of the ester group, which leads to the also diuretically active main metabolite (metabolite I) in the plasma of all 3 species; 2. glucuronidation of the resulting metabolite I, leading to metabolites II and III, which are diastereoisomeric esters of the two enantiomeric forms of metabolite I with beta-D-glucuronic acid. 50--60% of the urinary radioactivity can be described with these two metabolisation steps in all 3 species; 3. Oxidation of the piperidine moiety to metabolites IV--VII.