Systemic bioavailability and pharmacokinetics of the doxylamine-pyridoxine delayed-release combination (Diclectin).

BACKGROUND: Diclectin, composed of 10 mg doxylamine succinate (DOX) and 10 mg pyridoxine hydrochloride, is the drug combination of choice for the management of nausea and vomiting during pregnancy in Canada. However, there is large variability in its onset and duration of action among women. To understand and improve its effectiveness, the variability in the pharmacokinetics of the ingredients in this doxylamine succinate/pyridoxine hydrochloride combination must be studied. OBJECTIVES: To determine the pharmacokinetics of DOX and pyridoxine after oral administration of two tablets of this drug combination in the form of Diclectin and to calculate their respective relative bioavailability by comparison with intravenous administration in another population. METHODS: Eighteen nonpregnant, nonlactating, healthy females between 18 and 45 years of age were administered two tablets of Diclectin with 240 mL of water under empty stomach conditions. Blood samples were analyzed for DOX and pyridoxine along with its four active metabolites: pyridoxal, pyridoxal-5'-phosphate (PLP), pyridoxamine, and pyridoxamine-5'-phosphate using tandem mass spectrometry. For the purpose of this study, pharmacokinetic values for DOX and PLP were adjusted for body weight. RESULTS: The mean DOX-AUC0→∞ was calculated to be 3137.22 ± 633.57 ng·hr/mL (range, 2056.59-4376.06 ng·hr/mL). The mean PLP-AUC0→∞ was calculated to be 5513.10 ± 2362.35 ng·hr/mL (range, 1572.56-10,153.77 ng·hr/mL). Based on literature values of the PLP-AUC0→∞ after intravenous administration and data from the current study, the relative bioavailability of pyridoxine in Diclectin was calculated at 100%. CONCLUSION: There was a 2.1-fold variability in the DOX-AUC0→∞ and 6.5-fold variability in the PLP-AUC0→∞ after oral administration of 20 mg Diclectin. Using literature values and data from the current study, we estimated the oral bioavailability of pyridoxine to be 100%. Therefore, interindividual differences in metabolism, and not in bioavailability, may be important sources of variability that need to be addressed in dosing guidelines.

Manganese-Enhanced MRI of the Brain in Healthy Volunteers.

BACKGROUND AND PURPOSE: The manganese ion is used as an intracellular MR imaging contrast agent to study neuronal function in animal models, but it remains unclear whether manganese-enhanced MR imaging can be similarly useful in humans. Using mangafodipir (Teslascan, a chelated manganese-based contrast agent that is FDA-approved), we evaluated the dynamics of manganese enhancement of the brain and glandular structures in the rostral head and neck in healthy volunteers. MATERIALS AND METHODS: We administered mangafodipir intravenously at a rate of 1 mL/minute for a total dose of 5 µmol/kg body weight. Nine healthy adult volunteers (6 men/3 women; median age, 43 years) completed baseline history and physical examination, 3T MR imaging, and blood work. MR imaging also followed mangafodipir administration at various time points from immediate to 7 days, with delayed scans at 1-3 months. RESULTS: The choroid plexus and anterior pituitary gland enhanced within 10 minutes of infusion, with enhancement persisting up to 7 and 30 days, respectively. Exocrine (parotid, submandibular, sublingual, and lacrimal) glands also enhanced avidly as early as 1 hour postadministration, generally resolving by 1 month; 3 volunteers had residual exocrine gland enhancement, which resolved by 2 months in 1 and by 3 months in the other 2. Mangafodipir did not affect clinical parameters, laboratory values, or T1-weighted signal in the basal ganglia. CONCLUSIONS: Manganese ions released from mangafodipir successfully enable noninvasive visualization of intra- and extracranial structures that lie outside the blood-brain barrier without adverse clinical effects, setting the stage for future neuroradiologic investigation in disease.

Anticonvulsant characteristics of pyridoxyl-gamma-aminobutyrate, PL-GABA.

GABA is the major inhibitory neurotransmitter in the central nervous system, and its concentration in the brain in associated with a variety of neurological disorders, including seizures, convulsions, and epilepsy. The concentration of GABA is modulated by the pyridoxal-5'-phosphate (PLP)-dependent enzymes, GAD and GABA-T. In this study, we generated pyridoxyl-gamma-aminobutyrate (PL-GABA), a novel GABA analogue composed of pyridoxyl and GABA, and have also characterized its anticonvulsant and pharmacological functions in vitro. The results of biodistribution studies revealed that PL-GABA is capable of crossing the blood-brain barrier. PL-GABA evidenced anticonvulsant activity in a wide range of epilepsy models, some of which were electrically-based (MES seizures) and some chemically-based (bicuculline, pentylenetetrazol (PTZ), picrotoxine, 3-mercaptopropionic acid). Following a timed subcutaneous administration of PTZ to mice, PL-GABA consistently increased the latencies to first twitch and clonus. In addition, PL-GABA displayed no signs of tolerance after subchronic (10 day) treatment. PL-GABA appears to exert its anticonvulsant effects by influencing seizure spread and by raising the seizure threshold. Therefore, our results indicate that PL-GABA exerts a broad-spectrum anticonvulsant effect, and identify the potential for reduced PL-GABA tolerance as an additional positive profile for novel antiepileptic drugs.

Contrast enhanced cartilage imaging: Comparison of ionic and non-ionic contrast agents.

Our objective was to compare relaxation effects, dynamics and spatial distributions of ionic and non-ionic contrast agents in articular cartilage at concentrations typically used for direct MR arthrography at 1.5T. Dynamic MR-studies over 11h were performed in 15 bovine patella specimens. For each of the contrast agents gadopentetate dimeglumine, gadobenate dimeglumine, gadoteridol and mangafodipir trinatrium three patellae were placed in 2.5mmol/L contrast solution. Simultaneous measurements of T(1) and T(2) were performed every 30min using a high-spatial-resolution "MIX"-sequence. T(1), T(2) and DeltaR(1), DeltaR(2) profile plots across cartilage thickness were calculated to demonstrate the spatial and temporal distributions. The charge is one of the main factors which controls the amount of the contrast media diffusing into intact cartilage, but independent of the charge, the spatial distribution across cartilage thickness remains highly inhomogeneous even after 11h of diffusion. The absolute DeltaR(2)-effect in cartilage is at least as large as the DeltaR(1)-effect for all contrast agents. Maximum changes were 5-12s(-1) for DeltaR(1) and 8-15s(-1) for DeltaR(2). This study indicates that for morphologically intact cartilage only the amount of contrast agents within cartilage is determined by the charge but not the spatial distribution across cartilage thickness. In addition, DeltaR(2) can be considered for quantification of contrast agent concentrations, since it is of the same magnitude and less time consuming to measure than DeltaR(1).

Effects of consecutive high-dose alcohol administration on the utilization of sulfur-containing amino acids by rats.

In this study, we attempted to evaluate changes in sulfur-containing amino acid (SCAA) metabolism after short-term high-dose alcohol ingestion. At the beginning of the study, six animals were sacrificed as the baseline group and then other animals in the experiment were consecutively gavaged with alcohol (30%, 3 g/kg) for 7 days. Animals (n=6 each) were subsequently sacrificed at the time points of Days 1 (Group E1), 3 (Group E3) and 7 (Group E7). Blood samples and selected tissues were collected at each time interval. SCAA, pyridoxal phosphate (PLP) and glutathione (GSH) levels were analyzed. Results showed that taurine levels of tissues (brain, liver, heart and kidneys) all declined after the ethanol intervention and continued to decrease in selected tissues except the brain during the experiment. Furthermore, the trends of plasma taurine and PLP contents were highly correlated (r=.98, P=.045). A similar utilization pattern of plasma taurine and PLP indicated that transsulfuration preferred taurine production to GSH synthesis. The trend of plasma taurine levels being positively correlated with PLP levels reveals that dramatic transsulfuration occurred to meet the urgent demand for taurine by brain cells. In conclusion, we reported that continual alcohol ingestion alters SCAA utilization, especially by depletion of taurine and hypotaurine and by elevation of S-adenosyl homocysteine in the selected organs.

Kinetics of antagonist actions at rat P2X2/3 heteromeric receptors.

1. Currents through heteromeric P2X(2/3) receptors were evoked by applying alpha,beta-methylene-ATP to human embryonic kidney cells transfected with cDNAs encoding the P2X(2) and P2X(3) subunits. The concentration of alpha,beta-methylene-ATP were < or =30 microM because higher concentrations can activate homomeric P2X(2) receptors. The kinetics of action of three structurally unrelated antagonists were studied; these were 2', 3'-O-(2,4,6,trinitrophenyl)-ATP (TNP-ATP), pyridoxal-5-phosphate-6-azophenyl-2',4'-disulphonate (PPADS) and suramin. The association and dissociation rate constants were determined by pre-applying the antagonist for various periods prior to the co-application of agonist and antagonist, or by changing the solution from one containing only the agonist to one containing both agonist and antagonist. The high affinity of TNP-ATP for the P2X(2/3) receptor (K(D) approximately 2 nM) results from fast binding (k(+1) approximately 100 microM(-1) s(-1)) rather than slow unbinding (k(-1) approximately 0.3 s(-1)). For suramin (K(D) approximately 1 microM) the association rate constant ( approximately 1 microM(-1) s(-1)) was 100 times slower than that of TNP-ATP but the dissociation rate constant was similar (k(-1) approximately 1 s(-1)). PPADS (K(D) approximately 0.1 microM) associated and dissociated some 100 - 10,000 times more slowly than the other antagonists.

Effects of MnDPDP, DPDP--, and MnCl2 on cardiac energy metabolism and manganese accumulation. An experimental study in the isolated perfused rat heart.

RATIONALE AND OBJECTIVES: Recent studies indicate that manganese dipyridoxyl diphosphate (MnDPDP) may function as a slow release agent for manganese ions (Mn++) and that MnDPDP is approximately 10 times less potent than manganese chloride (MnCl2) in depressing cardiac function. The authors examined the possibility that MnDPDP and MnCl2 may influence cardiac metabolism and enzyme release and lead to a tissue accumulation of Mn. METHODS: Manganese DPDP, DPDP--, or MnCl2 (1000 microM) was infused in isolated rat hearts, which were freeze-clamped at various time intervals during infusion (5 minutes) and recovery (14-minute washout). Enzyme (lactate dehydrogenase) release, tissue high energy phosphates, Mn contents, and physiologic indices were measured at various time intervals. RESULTS: No significant differences were noted for: lactate dehydrogenase in the treated groups; tissue creatine phosphate (CrP) and adenosine triphosphate in MnDPDP, DPDP--, and control groups; and tissue Mn in DPDP-- and control groups. Manganese-chloride and MnDPDP-treated hearts accumulated and retained Mn in an 8:1 ratio. Manganese chloride depressed cardiac function more effectively than MnDPDP. CONCLUSIONS: The study has shown that: heart tissue uptake and retention of Mn++ is rapid and effective; MnCl2 is approximately eight times more potent than MnDPDP in promoting these effects; and a rise in tissue Mn content to eight to nine times (MnDPDP) or 60 to 70 times (MnCl2) the normal level does not lead to acute side effects on cardiac energy metabolism, function, and enzyme release. The study indicates that MnDPDP may act like a slow release compound for Mn++ ions.

Cardiac metal contents after infusions of manganese. An experimental evaluation in the isolated rat heart.

RATIONALE AND OBJECTIVES: Manganese dipyridoxyl diphosphate (MnDPDP), a contrast agent for liver MRI, releases free Mn2+ in a graded manner. The aim of the study was to compare the effects of brief versus prolonged infusions of MnDPDP and manganese chloride (MnCl2) on cardiac function, metabolism, Mn accumulation, and tissue metal content. METHODS: Isolated perfused rat hearts received 1-minute or 10-minute infusions of MnDPDP (100 microM, 1000 microM) or of MnCl2 (10 microM, 100 microM). Physiologic indices were measured intermittently, and tissue high-energy phosphate compounds and Ca/Fe/Mg/Mn/Zn contents were measured after a standardized Mn washout. RESULTS: One-minute and 10-minute infusions induced, respectively, minor and marked depressions of contractile function and corresponding elevations in myocardial Mn content. MnCl2 was markedly more potent than MnDPDP. Ten-minute infusions of the highest concentration of MnDPDP and MnCl2 lowered tissue Mg and elevated tissue Ca (MnCl2), whereas high-energy phosphates were unaffected. CONCLUSIONS: Mn uptake after Mn infusion is strongly related to the duration, concentration, and dose of free Mn ions. Differences in Mn accumulation between MnDPDP and MnCl2 were more pronounced after the 10-minute infusion.

Distribution of [3H]alpha,beta-methylene ATP binding sites in rat brain and spinal cord.

The distribution of [3H]alpha,beta-methylene ATP ([3H]alpha,beta-MeATP) binding sites in rat brain and spinal cord was examined using autoradiography. It was shown that many structures in the CNS are densely labelled, and that the binding is displaced by both beta,gamma-methylene ATP and the P2X-purinoceptor antagonist, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), indicating that the binding sites are P2X-purinoceptors. Image analysis revealed that the nuclei of the thalamus have the highest density of labelling. Other densely labelled structures include: the amygdaloid, substantia nigra, cerebral cortex, hypothalamus, caudate putamen, geniculate nuclei, medial habenula and the intermediate zone of grey matter in the spinal cord. These results are consistent with the limited amount of electrophysiological evidence for the existence of P2X-purinoceptors in the CNS, and provoke the need for extensive physiological studies to establish the roles of P2X-purinoceptors in the CNS.

The uptake of Mn-DPDP by hepatocytes is not mediated by the facilitated transport of pyridoxine.

Manganese-dipyridoxal diphosphate (Mn-DPDP) is a liver-selective contrast agent selectively taken up by the hepatocytes. Because of the analogy of structure with pyridoxine (vitamin B6), it was previously suggested that this compound can be selectively taken up by the facilitated transport of vitamers B6. To understand the uptake mechanism, an in vivo binding study was performed based on a competition between 54Mn-DPDP and pyridoxine on the one hand, and Mn-DPDP and [3H]pyridoxine on the other. We found that the [3H]pyridoxine levels in the liver were not significantly different 5 min after intravenous administration of several doses of Mn-DPDP (5 nmol/kg to 50 mumol/kg): 5.0 +/- 0.3% of the injected dose/g tissue. The content of 54Mn (administered as 54Mn-DPDP) in the liver was not affected by a saturation dose of pyridoxine (1 mmol/kg) and was found to be constant (+/- 10% of the injected dose/g tissue) for 60 min. These experiments showed that the uptake of Mn-DPDP is not mediated by the transporter of pyridoxine.

Role of exocrine cells in pancreatic enhancement using Mn-DPDP-enhanced MR imaging.

OBJECTIVE: To investigate role of exocrine cells in the pancreatic enhancement images at Manganese (II) N, N'-dipyridoxylethlenediamine-N, N'-diacetate 5, 5'-bisc (Mn-DPDP)-enhanced magnetic resonance (MR) imaging. METHODS: Artificial pancreatic leakage was constructed in six dogs using a fistula tube inserted into the duodenum papillae. Pancreatic juice was collected before and after intravenous infusion of 2 ml/kg of Mn-DPDP at a rate of 2 - 3 ml/min. The Mn content of pancreatic juice was measured by atomic absorption spectroscopy. T(1)-weighted spin-echo images and T(1)-weighted spoiled phase gradient-echo (SPGR) images were obtained prior and approximately 30 min after the administration of Mn-DPDP at 1.5T. RESULTS: The Mn content of pancreatic secretion increased 60.47 +/- 21.83 micro g/dl after the administration of Mn-DPDP (t = 6.785, P < 0.01). The signal/noise ratio (S/N) of the pancreas increased 53 percent +/- 49 percent and 62 percent +/- 44% on T(1)W spin echo images and SPGR images, respectively. CONCLUSIONS: Exocrine cells of the pancreas can absorb manganese and excrete it through the pancreatic juice. Exocrine cells play an important role in the enhancement of the pancreas in MR imaging with Mn-DPDP.

Intestinal absorption of pyridoxal 5'-phosphate at physiological levels in rats.

The intestinal absorption of pyridoxal 5'-phosphate (PLP) at physiological levels (10(-7) -10(-6) M) was studied in comparison with that of pyridoxal (PL) in rat, using in vitro everted sac and an intestinal preparation that permitted continuous in situ collection of mesenteric venous blood. After PLP administration (10(-6) -10(-3) M) in situ, larger amounts of PLP were found in the mesenteric venous plasma than after PL administration at the same dose. The amount of PLP found in the mesenteric venous plasma was dependent on its dose at lower concentrations up to 10(-4) M but became independent at higher concentrations. After PL administration at various doses, the amount of PL found in the mesenteric venous blood increased linearly with the dose. When various concentrations of PLP were added to the mucosal side, under the in vitro condition with protection from alkaline phosphate hydrolysis, PLP was detected in the serosal side and the extent of PLP transport was dependent on the initial concentration of PLP in the mucosal side. When various concentrations of PL were added to the mucosal side, the extent of PL transport was independent of the initial concentration of PL in the mucosal side. In rat pretreated with actinomycin D, PLP transport in vitro was inhibited but not that of PL. N2-induced anoxia and pyridoxamine 5'-phosphate and anion transport inhibitor (4,4'-diisothiocyanostilben-2,2'-disulfonic acid disodium salt) showed no effect on PLP transport. These results suggest that PLP can be absorbed in the phosphorylated form and imply the presence of a saturable process for direct absorption of PLP itself and a diffusive process for PL absorption. In addition, the result of the in vivo neonatal experiment suggests that the neonatal intestine also can transport PLP in phosphorylated form.

Some pharmacokinetic considerations about homeostatic equilibrium of endogenous substances.

Endogenous substances in the body are controlled through simple, very effective mechanisms, that preserve an optimum homeostatic equilibrium of baseline concentration and restore it when impaired. When planning a pharmacokinetic investigation of an endogenous substance exogenously administered, it is imperative to carefully ascertain the above mechanisms as well as the baseline value and their possible variations associated with daily rhythm, food, age, sex, menstrual cycle. Often the control mechanisms operate through non-linear processes, therefore a non-compartmental analysis or a tailored model may be more appropriate than the compartmental models used in standard pharmacokinetic analysis. Some specific examples of endogenous substances are discussed here on the basis of the data from the literature and personal experience.

[Metabolism and catabolism to CO2 of [14C] gamma-aminobutyric acid conjugates with pyridoxal phosphate, nicotinate and biotin]. / Obmen i katabolizm do CO2 kon"iugatov [14C]gamma-aminomaslianoi kisloty s piridoksal'fosfatom, nikotinatom i biotinom.

Pharmacokinetics and catabolism to CO2 in the mice organism and metabolic transformation by the tissues homogenates of [1-14C] GABA and its conjugates with nicotinate, pyridoxal phosphate and biotin have been studied. The permeability of nicotinoyl-GABA through the hemato-encephalic barrier was 10 times as much as the corresponding value for GABA and its conjugates with other vitamins. PLP-GABA is eliminated more rapidly from the brain in comparison with GABA, biotinyl-GABA is retained to a higher degree in kidneys, the entero-hepatic recycling takes place more actively for nicotinoyl-GABA. The latter, in contrast to biotinyl-GABA, remains unaffected by the liver, intestine mucose membrane and brain proteases and is catabolized to CO2 to a considerably lower extent as compared with GABA, perhaps due to the intestine bacterial microflora.

[Distribution of 1-[14C] GABA and its conjugates with nicotinate, pyridoxal phosphate and biotin in tissues of guinea pigs in closed systems]. / Rozpodil [1-14C] GAMK ta ii kon'iugativ z nikotynatom, pitydoksalfosfatom i biotynom u tkanynakh morskykh svynok pry dii faktoriv zamknenoho prostoru.

Distribution of the total radioactive label in the blood, different parts of the brain (the whole cortex and its lobes, bulbi olfactorii, hippocamp, striopallidum, hypothalamus, thalamus, peduculi cerebri, cerebellum, pons, medulla oblongata), eye (retina, hyaloid and vascular membrane), hypophysis and liver of guinea pig males has been studied after the hypodermic injection of [14C] GABA and products of its conjugation with pyridoxal phosphate, nicotinic acid and biotin in the equimolar dose (50 nmol/g) under normal conditions and in case of action of the closed volume factors. Accumulation of the total label in the hypophysis after injection of [14C] GABA, PLP-[14C] GABA and nicotinoyl-[14C] GABA was 3.6, 3.1 and 2.7 times (respectively) as much as that in the blood. The content of the labelled compounds in the cortical structures of the telencephalon and in the diencephalon was the greatest in comparison with other parts of the brain. The permeability through a hemato-ophthalmic barrier was 3-5 times as much as that through a hemato-encephalic barrier. The closed volume factors lead to higher accumulation of the label as compared with the control after injection of [14C] GABA, PLP-[14C] GABA and biotinyl-[14C] GABA.

Biomedical aspects of pyridoxal 5'-phosphate availability.

The biologically active form of vitamin B6, pyridoxal 5'-phosphate (PLP), is a cofactor in over 160 enzyme activities involved in a number of metabolic pathways, including neurotransmitter synthesis and degradation. In humans, PLP is recycled from food and from degraded PLP-dependent enzymes in a salvage pathway requiring the action of pyridoxal kinase, pyridoxine 5'-phosphate oxidase and phosphatases. Once pyridoxal 5'-phosphate is made, it is targeted to the dozens different apoenzymes that need it as a cofactor. The regulation of the salvage pathway and the mechanism of addition of PLP to the apoenzymes are poorly understood and represent a very challenging research field. Severe neurological disorders, such as convulsions and epileptic encephalopathy, result from a reduced availability of pyridoxal 5'-phosphate in the cell, due to inborn errors in the enzymes of the salvage pathway or other metabolisms and to interactions of drugs with PLP or pyridoxal kinase. Multifactorial neurological pathologies, such as autism, schizophrenia, Alzheimer's disease, Parkinson's disease and epilepsy have also been correlated to inadequate intracellular levels of PLP.

A randomized, double-blind, crossover study on the pharmacokinetics of a novel formulation of CoQ10 with pyridoxal 5'-phosphate and phosphatidyl choline.

The pharmacokinetics of a single 30-mg dose of a novel enteric-coated coenzyme Q10 (CoQ(10)) formulation with pyridoxal 5'-phosphate and phosphatidyl choline (CoQ(10)-P5P-PC) was investigated against two comparators CoQ(10) (NPN 02176955) and CoQ(10) (DIN 02231736) in 21 healthy volunteers, with screening CoQ(10) levels of 0.8 ± 0.2 mg/L. A randomized, double-blind, crossover study was designed with a washout period of 2 weeks between each formulation and blood sampled at 2, 4, 5, 6, 8, 12, 24, 48 and 72 hr postdose. Significantly, higher plasma concentrations were demonstrated for the CoQ(10) (NPN 02176955) formulation at 6 and 8 hr postdose (p = .010 and p = .042, respectively). There were no significant differences between formulations with respect to the area under the curve, AUC((0-72 hr)), or the maximum plasma concentration (C(max)). Total CoQ(10) (T(max)) reached maximum plasma concentrations at 6.4 ± 2.5 hr after supplementation with CoQ(10) (NPN 02176955), 8.0 ± 9.8 hr with CoQ(10)-P5P-PC, and 9.5 ± 9.3 hr with CoQ(10) (DIN 02231736). The estimated elimination half-life (t(1/2)) was 92.3 hr after a single oral dose of CoQ(10)-P5P-PC, 38.2 hr with CoQ(10) (NPN 02176955), and 80.7 hr with CoQ(10) (DIN 02231736). The results suggest that CoQ(10) is available for a longer time in subjects' administered with CoQ(10)-P5P-PC in comparison with the other two formulations studied. There were no significant differences in adverse events, by severity, causality, or organ system. The CoQ(10)-P5P-PC formulation was found to be superior in the t(1/2), and it may be suggested that fewer doses are required to maintain healthy circulatory CoQ(10) levels.