Comparison of bioavailability of two ubidecarenone products in healthy Korean volunteers.

OBJECTIVE: This study aimed to evaluate the bioavailability of two pharmaceutical products of ubidecarenone (coenzyme Q10, CoQ10). MATERIALS: Two brands (brand A and brand B) of commercial CoQ10 hard capsules. METHODS: Two brands of CoQ10 capsules were administered at 100 mg dose to two groups of healthy volunteers, respectively, and blood samples were withdrawn at predetermined time intervals and assayed by a validated HPLC method with an electrochemical detector. RESULTS AND CONCLUSIONS: Intra- and inter-day precision and inter-day accuracy were acceptable for all quality control samples including the lower limit of quantitation of 50 ng/ml. Recovery of CoQ10 from human plasma was greater than 98.2%. CoQ10 was stable in human plasma under various storage conditions. This method was applied to a pharmacokinetic study after oral administration of CoQ10 hard capsules to healthy volunteers. The intrinsic CoQ10 concentrations were measured for three consecutive days before drug administration, which were ranged between 0.68 and 0.79 microg/ml, and there was no statistically significant difference between groups. In brand A, the plasma concentration after administration of CoQ10 was not higher than the intrinsic level, indicating that no significant drug absorption occurred, whereas considerably higher concentrations were obtained with brand B. The dissolution rates of brand A and B after 3 h were 0.35 +/- 0.09 and 1.27 +/- 0.16%, respectively. From the adjusted concentration-time curve, the AUC and t1/2 of brand B were calculated to be 11.51 +/- 5.76 microg x h/ml and 21.7 h, respectively. A mean Cmax of 0.32 +/- 0.1 microg/ml was obtained at 7.9 h. In conclusion, it was found that bioavailability of CoQ10 was significantly different depending on the formulations, and dissolution could be one of the important factors affecting CoQ10 absorption.

Analgesic effects of intra-nasal enkephalins.

The analgesic effects of intranasal delivery of leucine enkephalin (Leu-Enk) and its synthetic analogue [D-ala(2)]-leucine enkephalinamide (YAGFL) with or without enzyme inhibitors and/or absorption enhancers were investigated using the acetic acid-induced writhing test in mice. The analgesic activity was significantly affected by the time delay after the administration of Leu-Enk; the inhibition rates for the groups administered with acetic acid 5 min and 30 min after the administration of Leu-Enk were 56.40 +/- 8.54 and 17.98 +/- 7.07%, respectively. The addition of enzyme inhibitors and absorption enhancers markedly increased the inhibition rate of Leu-Enk and YAGFL; their inhibition rates were about four times and twice those without any enzyme inhibitor or absorption enhancer, respectively. The enzyme inhibitors and absorption enhancers that produced the highest inhibition rates of Leu-Enk and YAGFL were azelaic acid (1%), thimerosal (0.5 mM, TM), ethylenediamine-tetraacetic acid (5 mM, EDTA) and L-alpha-lysophosphatidylcholine (0.5%, LPC), and TM (0.5 mM), EDTA (5 mM), LPC (0.5%) and povidone (5%), respectively. The ED50 value of both enkephalins was also determined and found to be about 13 microg kg(-1), which is 850 and 60 times more potent than literature values for ketoprofen and morphine, respectively. Based on these results it was concluded that Leu-Enk or YAGFL could exert very high analgesic activity when administered nasally with a combination of inhibitors and absorption enhancers as compared with other analgesics.

Development of a method for the determination of human skin moisture using a portable near-infrared system.

In this study, a portable near-infrared (NIR) system was newly integrated with a photodiode array detector that has no moving parts, and this system has been successfully applied for the evaluation of human skin moisture. The good correlation between NIR absorbance and the absolute water content of separated hairless mouse skin, in vitro, was showed, depending on the water content (7.4-84.9%) using this portable NIR system. Partial least squares (PLS) regression was used for calibration with the 1150-1650-nm wavelength range. For practical use for the evaluation of human skin moisture, the PLS model for human skin moisture was developed in vivo using the portable NIR system on the basis of the relative water content values of stratum corneum from the conventional capacitance method. The PLS model showed a good correlation. This study indicated that the portable NIR system, as compared to conventional methods, could be a powerful tool for human skin moisture, which may be much more stable to environmental conditions, such as temperature and humidity. Furthermore, to confirm the performance of the newly integrated portable NIR system, a scanning-type conventional NIR spectrometer was used in the same experiments, and the results were compared.

Effect of vehicles and enhancers on the in vitro skin penetration of aspalatone and its enzymatic degradation across rat skins.

The feasibility of skin penetration was studied for aspalatone (AM, acetylsalicylic acid maltol ester), a novel antithrombotic agent. In this study, hairless mouse dorsal skins were used as a model to select composition of vehicle and AM. Based on measurements of solubility and partition coefficient, the concentration of PG that showed the highest flux for AM across the hairless mouse skin was found to be 40%. The cumulative amount permeated at 48 h, however, appear inadequate, even when the PG concentration was employed. To identify a suitable absorption enhancer and its optimal concentration for AM, a number of absorption enhancers and a variety of concentration were screened for the increase in transdermal flux of AM. Amongst these, linoleic acid (LOA) at the concentration of 5% was found to have the largest enhancement factor (i.e., 132). However, a further increase in AM flux was not found in the fatty acid concentration greater than 5%, indicating the enhancement effect is in a bell-shaped curve. In a study of the effect of AM concentration on the permeation, there was no difference in the permeation rate between 0.5 and 1% for AM, below its saturated concentration. At the donor concentration of 2%, over the saturated condition, the flux of AM was markedly increased. A considerable degradation of AM was found during permeation studies, and the extent was correlated with protein concentrations in the epidermal and serosal extracts, and skin homogenates. In rat dorsal skins, the protein concentration decreased in the rank order of skin homogenate > serosal extract > epidermal extract. Estimated first order degradation rate constants were 6.1 5 +/- 0.14, 0.57 +/- 0.02 and 0.011 +/- 0.004 h(-1) for skin homogenate, serosal extract and epidermal extract, respectively. Therefore, it appeared that AM was hydrolyzed to some extent into salicylmaltol by esterases in the dermal and subcutaneous tissues of skin. Taken together, our data indicated that transdermal delivery of AM is feasible when the combination of PG and LOA is used as a vehicle. However, since AM is not metabolically stable, acceptable degradation inhibitors may be necessary to fully realize the transdermal delivery of the drug.

In vitro percutaneous absorption of tenoxicam from pressure-sensitive adhesive matrices across the hairless mouse skin.

To investigate the feasibility of developing a new tenoxicam plaster, the effects of vehicles and penetration enhancers on the in vitro permeation of tenoxicam from a pressure-sensitive adhesive (PSA) matrices across the dorsal hairless mouse skin were studied. Vehicles employed in this study were propylene glycol (PG)-oleyl alcohol (OAI), PG-oleic acid (OA), and diethylene glycol monoethyl ether (DGME)-propylene glycol monolaurate (PGML) cosolvents with/without fatty acids. In this study, amines such as triethanolamine (TEA) and tromethamine (TM) were additionally used as a solubilizer. Among PSAs used, Duro-Tak 87-2510 showed much higher release rate than either Duro-Tak 87-2100 or Duro-Tak 87-2196. The relatively high flux rate was obtained with the formulation of DGME-PGML (40:60, v/v) with 3% OA and 5% TM, and the flux increased as a function of the dose; the initial flux up to 12 h was 4.98 +/- 1.38 microg/cm2/h at the tenoxicam dose of 50 mg/70 cm2. This flux was much higher than that of a commercial piroxicam patch (Trast) (1.24 +/- 0.73 microg/ cm2/hr) with almost only one-third that of the commercial patch. Therefore, these observations indicated that these composition of tenoxicam plaster may be practically applicable.

Age-related changes of water content in the rat skin.

The water content of the skin is greatly influenced by ground substances, which may be responsible for wrinkling and laxity of the skin accompanying the cutaneous aging. Therefore, water content in the skin is presumed to be a critical determinant in cutaneous aging. This study was aimed at clarifying the change in water content and the content of glycosaminoglycans (GAG) of rat skin in relation to aging. Sprague-Dawley rats were divided into 6 groups: 1-, 3-, 6-, 12-, 18- and 24-month-old groups. Two-to-three grams of skin tissue samples were taken from the back, and a half of sample was dried at 160 degrees C for 30 min with electronic moisture balance, and water content was assessed as decreased weight by heating. To measure change of GAG of the rat skin, another half of samples were extracted with 0.1 M sodium phosphate buffer (pH 7.4 NaPB) and 2 M guanidine-HCl/Tris buffer (pH 7.4). The resultant insoluble pellet was dried at 50 degrees C in a drying over for 72 h after two washings and the dry weight was recorded. The amount of sulfated GAG in the skin extracts was measured by alcian blue precipitation assay, and the amount of uronic acid (UA) was assayed in the skin tissue extracts and the dried skin using the carbazole reaction. The water content of the rat skin decreased with age, and a similar decreasing pattern in the amount of sulfated GAG and UA of the rat skin tissue was observed with aging. One hundred times of UA was obtained in dry rat skin tissue, as compared with that of the skin extracts. In conclusion, there occurs a significant decrease of water content in the aged rat skin, which may be related to the change of GAG with intrinsic aging of the skin.

Effects of flavonoids of Ginkgo biloba on proliferation of human skin fibroblast.

Ginkgo biloba studies have focused on the anti-inflammatory effects of the major components, ginkgolide and bilobalide, whereas little is known about their effect on fibroblasts. This study demonstrated the enhancing effects of Ginkgo L. extracts, especially the flavonoid fractions: quercetin, kaempferol, sciadopitysin, ginkgetin, isoginkgetin, on the proliferation of normal human skin fibroblast in vitro measured by MTT (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl-tetrazolium bromide) assay and direct hemocytometer cell count. Furthermore, increased production of collagen and extracellular fibronectin were documented by radioisotope (2,3-3H-proline) incorporated collagen assay, procollagen type I C-peptide assay and by immunoturbidimetric assay. These proliferative effects suggest another useful pharmacologic application of Ginkgo L. extracts in addition to their well-known anti-inflammatory effect.

A case of actinic granuloma associated with periumbilical perforating pseudoxanthoma elasticum.

We report an unusual case of actinic granuloma of the face and periumbilical perforating pseudoxanthoma elasticum located superior to the umbilicus in a 57-year-old Korean woman. Histopathologically, these two dermatoses have a similar degeneration of elastic fibers, but they show different host reactions to the altered elastic fibers. In the actinic granuloma, actinically damaged elastic fibers were followed by granulomatous infiltration on the sun-exposed area, while in the perforating pseudoxanthoma elasticum, the altered elastic fibers induced a foreign body reaction, with subsequent transepidermal elimination. This is the first case report showing both actinic granuloma and periumbilical perforating pseudoxanthoma elasticum in the same patient, which suggests that the basic mechanism eliciting these dermatoses is similar.

Transmucosal delivery of leucine enkephalin: stabilization in rabbit enzyme extracts and enhancement of permeation through mucosae.

Leucine enkephalin (Tyr-Gly-Gly-Phe-Leu; Leu-Enk) is a naturally occurring peptide that has been shown to have pain modulating properties. To evaluate the feasibility of using various absorptive mucosae as a route of systemic delivery, the stability of Leu-Enk and the effect of enzyme inhibitors (e.g., amastatin, EDTA, and thimerosal) on stabilization and permeation of Leu-Enk through rabbit mucosae in the presence of dihydrofusidates were investigated. Enzymes in the nasal, rectal, and vaginal mucosae were extracted and Leu-Enk (50 micrograms/mL) was added to each of the enzyme extracts and incubated to determine the kinetics and mechanism of degradation. The rate of degradation in the extracts in the absence of inhibitors followed the order: rectal > vaginal > nasal. Whereas EDTA had the best stabilizing effect on Leu-Enk, thimerosal was the best stabilizer for the degradation intermediates. A combination of amastatin (50 microM), EDTA (5 mM), and thimerosal (50 microM) had the greatest stabilizing effect on Leu-Enk and its degradation intermediates. For permeation studies, each mucosa was mounted onto a Valia-Chien permeation cell with Leu-Enk (200 micrograms/mL) in isotonic phosphate buffer (as donor solution). The enhancers used for the study were sodium tauro-dihydrofusidate (STDHF), sodium glycodihydrofusidate (SGDHF), and phosphato-dihydrofusidate (PHDHF). The greatest effect was achieved by PHDHF for all the mucosae. STDHF had a significant effect only on the rectal permeation, whereas SGDHF had significant effects on rectal and vaginal mucosae. Mechanisms by which the dihydrofusidates enhance permeating may involve micelle formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Transmucosal delivery of methionine enkephalin. I: Solution stability and kinetics of degradation in various rabbit mucosa extracts.

To evaluate the feasibility of transmucosal delivery of methionine enkephalin (Tyr-Gly-Gly-Phe-Met; Met-Enk), it is important to first investigate its physicochemical and enzymatic stability. The kinetics of degradation of Met-Enk in aqueous solution was determined at pH 2.01-9.84 and 37-45 degrees C by high-performance liquid chromatography. The first-order rate constant (k) was calculated, and the log k-pH profile showed that Met-Enk is most stable at pH approximately 5.0. Various mucosae excised from rabbit were mounted on Valia-Chien permeation cells and exposed to isotonic phosphate buffer at physiologic pHs. Mucosal and serosal extracts were collected from the donor and receptor solutions, respectively. The degradation of Met-Enk in the extracts followed first-order kinetics, but no significant difference in the degradation rates was observed between mucosal and serosal extracts, regardless of the type of mucosa used. Degradation was most rapid in the extracts of rectal mucosa, followed by vaginal and nasal mucosae. The major metabolites were Des-Tyr-Met-Enk and Tyrosine (Tyr), indicating the enzymatic hydrolysis by aminopeptidases. However, the data also suggested that dipeptidyl peptidase and dipeptidyl carboxypeptidase could play some roles in the degradation of Met-Enk. The degradation pathways of Met-Enk were further explored by concomitantly determining the formation of smaller metabolites of primary hydrolytic fragments of Met-Enk in the mucosal extracts.