Pharmacokinetics of aniracetam and its metabolites in rats.

The pharmacokinetics of aniracetam (AP), a new cognitive performance enhancer, and its main metabolites was investigated after intravenous (iv) and oral administrations to rat. The plasma levels of AP, 4-p-anisamidobutyric acid (ABA), and p-anisic acid (AA) were determined simultaneously by the HPLC method. The plasma concentrations of the parent drug and ABA quickly declined in a biexponential manner, with rapid terminal decay and a small mean residence time. However, AA yielded nonlinearly high levels at the initial times and the plasma concentrations of 2-pyrrolidinone (PD) were sustained over a relatively long time. When AA was administered intravenously, nonlinearity of the plasma concentrations was also found at higher doses. To describe the time course of the plasma levels of AP and its metabolites after iv administration, a pharmacokinetic model with seven compartments was applied, which included 10 first-order rate constants and one Michaelis-Menten constant. An approximate fit was obtained between the observed and calculated curves based on the model, except for the plasma concentrations of ABA. The plasma concentration-time profiles of AP and its metabolites following oral administration of AP (50 and 100 mg/kg) were similar to those after iv dosing, with the exception of PD, which showed much lower plasma levels than those after iv administration. Elimination of AP and ABA was rapid after oral dosing, and the bioavailability of AP was extremely small (11.4 and 8.6%). As a result, AP was largely metabolized to ABA, AA, and PD in rat.

Phase transitions of rat stratum corneum lipids by an electron paramagnetic resonance study and relationship of phase states to drug penetration.

In order to relate barrier function to stratum corneum structure and the thermal transitions of corneum lipids, samples from hairless rat skin were investigated by using ESR and drug penetration techniques. The phase transition of stratum corneum lipids was estimated using a deeper probe (16-doxyl-stearic acid) inserted in the lipid bilayers and measuring the rotational correlation time, tau(c). Results of ESR study showed that stratum corneum lipids underwent thermal transitions at 39.3 +/- 1.6 degrees C and 63.6 +/- 2.6 degrees C roughly similar to the data obtained by differential scanning calorimetry measurements. Cholesterol oxidase treatment decreased the fluidity of the lipids at lower temperatures. The treatment of stratum corneum with laurocapram (1%) and isopropyl myristate (IPM, 2%) little changed both phase transition temperatures, although the treatment highly increased the molecular motion of the lipids. The flux (J(s)) of lipophilic drugs (beta-estradiol, indomethacin and betahistine) through the skin was enhanced with increasing temperatures, with an increase in the diffusion constant within skin and a decrease in the lag time. There was a good relationship between log J(s) or log permeability coefficient (K(p)) and 1/tau(c) in the temperature range of 45 to 64 degrees C. The calculated activation energy (delta E) for diffusion of these drugs across skin was 17-40 kcal/mol. Judging from our data, stratum corneum lipids of rat probably exist as the gel, crystalline state below 39 degrees C, the mesomorphic state between 39 and 64 degrees C and the fluid, liquid-crystalline state at temperatures of 64 degrees C or above. These results are in line with the permeability of these lipophilic drugs through the intercellular lipids disordered is highly increased.

Percutaneous penetration of fluorescein isothiocyanate-dextrans and the mechanism for enhancement effect of enhancers on the intercellular penetration.

To identify the mechanism involved in the enhancement effect of enhancers on the intercellular penetration of large polar molecules, the skin penetration of fluorescein isothiocyanate (FITC)-dextrans (average molecular weight; 4400, 9400, and 69000 Da) and the lipid removal from the intercellular spaces by enhancers were studied using hairless rat skin. Pretreatment of hairless rat skin with enhancers such as n-octanol (20%), laurocapram (2%), isopropylmyristate (IPM, 20%), oleic acid (5%) and cineol (2%), which are water-immiscible, significantly enhanced the flux of FITC-dextrans, while pretreatment with water-miscible enhancers, i.e. dimethyl sulfoxide (DMSO, 5%) and N-methyl-2-pyrrolidone (NMP) did not increase the flux compared with the control. The penetration of FITC-dextrans was approximately size dependent. n-Octanol, laurocapram, IPM and oleic acid dramatically removed ceramides which are the intercellular lipids, whereas NMP and DMSO partly extracted the sphingolipids. A linear relationship was observed between the flux and removal of ceramides (p < 0.01), indicating that the removal of intercellular lipids would cause dramatic dilations between adherent cornified cells and enhance the penetration through the intercellular pathways. When the penetration of FITC-dextrans through Wistar rat skin was compared with that via hairless rat skin, the steady state flux of FITC-dextrans through Wistar rat skin pretreated with water-immiscible enhancers was 1.2- to 4.9-fold higher, suggesting that the penetration of large polar molecules through follicles may play at least some role in the percutaneous absorption.

Effect of various enhancers on transdermal penetration of indomethacin and urea, and relationship between penetration parameters and enhancement factors.

The enhancing capacity of various chemicals, which are widely recognized as enhancers, for the transdermal penetration into full-thickness rat skin of a model lipophilic drug [indomethacin (IND)] and a hydrophilic permeant (urea) was estimated by an in vitro technique. In addition, the fluidity of the stratum corneum lipids, the partitioning of IND into skin, the lipid (ceramides) extraction from the stratum corneum by enhancers, and the IND solubility in enhancer vehicle were measured and related to the enhancing capacity. In vitro permeation experiments with hairless rat skin unequivocally revealed that the enhancers varied in abilities to enhance the fluxes of both agents. Laurocapram, isopropylmyristate (IPM), sodium oleate, and cineol increased fluxes of both agents to a great extent, but N-methyl-2-pyrrolidone (NMP), N,N-diethyl-m-tolamide (DEET), and oleyl oleate were less effective acclerants. Many enhancers increased the fluidity of the lipids [with a threshold of approximately 0.6-0.8 ns at 37 degrees C in the rotational correlation time (tau c)], the skin partitioning of IND, the extraction of ceramides from the cornified cells, and the thermodynamic activity of IND in vehicle (calculated from the solubility) to varying extents. A good correlation was observed between the increase in the fluidity of stratum corneum lipids and the partitioning of IND into skin, between the increase in the fluidity and the flux or the decrease in lag time for IND, between the removal of ceramides and the skin partitioning of IND, and between the removal of ceramides and the flux of urea (p < 0.05 in all cases).(ABSTRACT TRUNCATED AT 250 WORDS)

Mechanism of the enhancement effect of n-octyl-beta-D-thioglucoside on the transdermal penetration of fluorescein isothiocyanate-labeled dextrans and the molecular weight dependence of water-soluble penetrants through stripped skin.

To clarify the mechanism of the enhancing effect of n-octyl-beta-D-thioglucoside (OTG), which acts as a potent enhancer for skin penetration of peptides and water-soluble penetrants, the in vitro penetration of macromolecules [fluorescein isothiocyanate-labeled dextrans (FTIC-dextrans)] was evaluated with hairless rat skin and stripped skin. The FITC-dextrans (MW, 4400, 9600, and 69,000 Da, referred to as FD-4, FD-10, and FD-70, respectively) penetrated more easily in the presence of OTG (1.5%), with high fluxes equivalent to those through stripped skin. This result indicated that the enhancement effect of OTG on the penetration of macromolecules through the stratum corneum was extensive, and the barrier function of the corneum was nearly eliminated by the OTG treatment. OTG significantly solubilized the stratum corneum proteins and ceramides during the initial time stage. Scanning electron microscopic observations demonstrated that OTG treatment dramatically changed the cell membrane (i.e., exfoliation of cell membranes and dissociation of adherent cornified cells), suggesting a significant disturbance of the cohesive laminae and barrier functions. The extent of dissociation of cell membranes increased with treatment time, without significant changes in the cell junctions. These results clarify that the enhancement mechanism of OTG was different from that of laurocapram and other lipophilic enhancers. The permeability of polar solutes with differing molecular sizes (MW, 180-69,000 Da) through stripped skin was size dependent (r = 0.997, p < 0.001). However, the viable epidermis and dermis restricted the penetration of macromolecules, such as FD-70.

Percutaneous absorption of physiologically active peptides, ebiratide and elcatonin, in rats.

This study was designed to evaluate the percutaneous absorption of physiologically active peptides, ebiratide (a behaviorally potent adrenocorticotropic analog) and elcatonin (a hypocalcemic peptide) in an attempt to develop an efficient transdermal therapeutic system for the treatment of diseases. The [125I]ebiratide penetration through rat skin from gel formulations could be described fairly well by a zero-order kinetic profile. Skin penetration was the greatest when EDTA, n-octyl-beta-D-thioglucoside (OTG, 1.5%) and taurocholate (1.0%) were combined in a gel formulation. The order of flux was: EDTA, OTG and taurocholate (formulation 3) > OTG and taurocholate (formulation 2) > glucosyl-beta-cyclodextrin and OTG (formulation 4). When the transdermal systems of [125I]ebiratide prepared using a corresponding gel formulation were applied to rat abdomen, the plasma levels of radioactivity after formulations 3 and 2 were much higher than those after formulation 1 without enhancers, and the radioactivity was observed in the brain, although in a very small quantity. The hypocalcemic effect of elcatonin was measured in vivo after application of the transdermal systems. The plasma calcium levels decreased comparatively rapidly and low levels were maintained for a long period, indicating the effectively percutaneous absorption of elcatonin. Formulation 7 containing D-limonen and taurocholate as enhancers and/or inhibitors showed much higher hypocalcemic effect than two other formulations combined with laurocapram or N,N-diethyl-m-toluamide, consequently giving the highest pharmacological availability (8.7 +/- 1.0%). These results clearly demonstrated that the peptides were effectively absorbed through rat skin in the presence of enhancers.

Pharmacokinetics of indomethacin octyl ester (prodrug) and indomethacin produced from the prodrug.

A prodrug of indomethacin, indomethacin octyl ester (IM-OE), was synthesized and its pharmacokinetics was investigated in rat. To describe the time course of the plasma indomethacin and IM-OE after intravenous (iv) and oral administrations, a pharmacokinetic model with four compartments was developed. Indomethacin rapidly appeared in plasma after iv administration of IM-OE and declined in a monoexponential manner, with a rapid decline and low plasma levels of IM-OE. The plasma concentrations of indomethacin after oral administration of IM-OE were much lower than those after oral administration of indomethacin. The high concentrations of IM-OE compared with indomethacin were detected in liver 3 h after oral dosing of the prodrug, although IM-OE was not detected in plasma. A good fit was obtained between the observed and calculated curves based on the model, which includes a conversion rate constant of IM-OE to indomethacin for both iv and oral dosings of IM-OE. Additionally, the model could successfully describe the plasma concentration versus time profiles after indomethacin dosings.

Effectiveness of the elcatonin transdermal system for the treatment of osteoporosis and the effect of the combination of elcatonin and active vitamin D3 in rat.

The efficacy of percutaneous elcatonin (EC), a hypocalcemic peptide, in the treatment of experimental osteoporosis in rats was evaluated in vivo. Additionally, the effect of the combined use of EC and active vitamin D3 (1,25(OH)2D3) for the treatment was compared with those of three other groups: 1,25(OH)2D3 alone, estradiol plus 1,25(OH)2D3, and a placebo, and low calcium diet (low Ca). The EC transdermal system and the EC plus 1,25(OH)2D3 system, applied to the rat abdominal skin 6 times for 48 h, significantly increased the ash weight and calcium content of the tibia in the rats, compared with those of placebo group (p < 0.05). The EC systems also slightly lowered the alkaline phosphatase activity in plasma of the morbid rats, without a difference in the plasma calcium content. These EC systems were superior to the 1,25(OH)2D3 system and the estradiol plus 1,25(OH)2D3 system in improving osteoporotic parameters. Thus, the EC systems were concluded to be an efficient drug delivery system for Paget's disease and osteoporosis.

Rectal absorption of acyclovir in rats and improvement of absorption by triglyceride base.

The rectal absorption of acyclovir has been evaluated after administration of suppositories without absorption enhancers in rats. The disappearance of plasma acyclovir followed biexponential kinetics after i.v. dosing. Rectal administration of a triglyceride (Vosco S-55 and Vosco S-55 + methylcellulose) suppository gave relatively high plasma concentrations and bioavailabilities (95.3 and 83.4%, respectively) compared with Witepsol and macrogol suppositories. However, the in vitro release profiles from suppositories did not accurately reflect plasma concentrations after rectal dosing. Our results suggest that the rectal administration of acyclovir suppositories may be a promising substitute for intravenous infusion, which is at present used for the treatment.