• Journal of Nutrition and Health
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• v.29 no.2
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• pp.153-158
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• 1996

Polyethylene glycols(PEGs)are hydrophilic molecules that have been used to characterize intestinal permeability via the paracellular pathway. Using a mixture of PEGs(400, 600 and 1000), containing oligomers in the molecular weight range 330 to 1122 D, the molecular weight permeability dependence in the jejunum of the rat small intestine was examined, employing an in situ recirculation perfusion technique. Individual oligomers were determined by HPLC with refractive detection. In the range studied, a distinct molecular weight cut-off was not apparent. Corrected for the length of jejunum used in the study, over the molecular weight range 330 to 1122D, the apparent permeability(Papp) of PEG ranged from 4.92$\pm$0.02$\times$10-5cm/sec(mean$\pm$SEM, n=5) to 0.28$\times$10-5cm/sec. Also, it was observed that the apparent permeability was inversely proportional to approximately MW2. The results in this study suggest that molecular weight is an important factor in determining the intestinal permeability.

• Archives of Pharmacal Research
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• v.6 no.2
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• pp.109-114
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• 1983

A mechanism of taurine transfer across the rat small intestine was elucidated by using the in situ recirculation perfusion or loop method. Taurine uptake was saturable, Km= 39.9 mM, and energy dependent, and required sodium. The close structural analogues, aminomethane sulfonic acid, .gamma.-amino-butyric acid, hypotaurine, and .betha.-alanine, reduced significantly taurine uptake when present in 10-fold excess. The .alpha.-amino acid, glycine, did not inhibit uptake. Hence, all of these findings lead to a conclusion that a carrier-mediated transport system for taurine exists in the small intestine.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.88-89
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• 2002

Traditionally, kinetics of brain influx of drugs has been evaluated by a number of experimental techniques. Brain uptake index and in situ brain perfusion study have been used for the determination of the kinetics; However, these methods generally focus on the accuracy of the uptake rate into the brain rather than the speed of the determination. In addition, application of radiolabelled substrates (e.g., $_{14}$C-labelled sucrose) further impedes the wide spread acceptance of these techniques for the application of high throughput screening system. (omitted)

• Archives of Pharmacal Research
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• v.17 no.5
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• pp.287-293
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• 1994

The effect of several adsorbents on paraquat poisohing was investigated (1) by measuring the saturatd amount of the poison adhered on the adsorbents in vitro and (2) by assaying the blood level of paraquat in the rat in in situ intestinal absorption experiments. Activated charcoal powder, natural aluminum silifonate) were used as adorbents. The steady-state blood level of paraquat in its absorption experiment with the cationic exchange resins was markedly lower than those without the resins or with other adsorbents. A good relationship was achieved between the calculated AUC or adsorptioin rate (in situ) and the saturated adsorption amount (in vitro). The rank order of the effect was sodium polystyene sulfonate > calcium polystyene sulfonate>natural aluminum silicate>activated charcoal powder. The effect of sodium polystyrene sulfonate after intestinal washing with physiological saline ws also measured, and a synergistic effect (marked decrease in blood paraquat level0 was found as compared with the intestinal washing alone. The simultaneous use of G.I. weshing and powerful adsorbent was scientifically proven to be most benefical.

• Journal of Pharmaceutical Investigation
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• v.21 no.4
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• pp.215-222
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• 1991

The absorption characteristics of itraconazole, which is an antifungal agent, from intestinal segments in the anesthetized rat i1l situ were investigated in order to design an effective oral drug delivery system. The pH-solubility profile of itraconazole, the rate and extent of absorption of itraconazole, the optimal absorption site(s) of itraconazole and the absorption enhancing effect of sodium cholate on itraconazole were examined in the present study. In situ single-pass perfusion method and recirculating perfusion technique using duodenum(D), jejunum(J) and ileum(I) were employed for the calculation of apparent permeability(Pe) and apparent first-order rate constant(Kobs). respectively. The results of this study were as follows; (1) Itraconazole showed appreciable aqueous solubility only at pH values of below 2.0. (2) pe(cm/sec) decreased in the following order: $D(10.24{\pm}1.78{\times}10^{-4})>J(8.86{\pm}0.79{\times}10^{-4})>I(3.78{\pm}0.13 X 10^{-4})$. (3) $Kobs(min^{-1})$ decreased in the following order: $J(17.12{\pm}3.19{\times}10^{-3})>D(13.37{\pm}0.6{\times}10^{-3})>I(11.05{\pm}0.91{\times}10^{-3})$. (4) The solubility of itraconazole markedly increased with the increase of the concentration of sodium cholate. (5) The addition of 10 mM sodium cholate significantly increased the apparent first-order rate constant of itraconazole in the ileum by a factor of 6.8.

• Journal of Pharmaceutical Investigation
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• v.24 no.2
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• pp.95-104
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• 1994

Inclusion complexes of ketoconazole(KT) with ${\alpha}^_$, ${\beta}^_$cyclodextrin(CD) and $dimethy1-{\beta}-cyclodextrin$ (CD) and $dimethy1-{\beta}-cyclodextrin(DM{\beta}CD)$ as nasal absorption enhancer were prepared in 1: 2 molar ratios by freeze-drying and solvent evaporation methods. In order to compare with the intrinsic absorptivity of KT in the jejunum(J) and the nasal cavity(N), the in situ simultaneous perfusion method was employed. The in situ recirculation study revealed that KT-CD inclusion complexes with the greater stability constant and the faster dissolution rate proportionally increased the absorption of KT in the J and N of rats. The rank order of apparent KT permeability$(P_{app}\;:\;cm/sec\;{\time}\;1O^{-5}{\pm}S.E.)$, corrected by surface area of absorption, was $5.10{\pm}0.3(N,\; KT-DM{\beta}CD)$ )> $4.13{\pm}0.4(N,\;KT-{\beta}-CD)$ )> $3.52{\pm}0.2(N,\;KT-{\alpha}-CD)$ )> $2.76{\pm}0.3(J,\; KT-DM{\beta}CD)$ )> $2.61{\pm}0.5(J,\;KT-{\beta}-CD)$ )> $2.42{\pm}0.4(J,\;KT-{\alpha}-CD)$ at pH 4.0. The in crease in permeability of $KT-DM{\beta}CD$ inclusion complex was 2.6 folds in the J and 4.5 folds in the N when the perfusing solution was changed from the buffer(pH 4.0) to saline. The absorption rate of $KT-DM{\beta}CD$ inclusion complex after nasal administration was more rapid than those of ketoconazole alone and $KT-DM{\beta}CD$ inclusion complex after oral administration to rats. In comparision with an oral administration of ketoconazole suspension in corn oil, the relative bioavailability was calculated 137.3% for the oral and 195.0% for nasal $KT-DM{\beta}CD$ inclusion complex in rats. The present results suggest that $KT-DM{\beta}CD$ inclusion complex may serve as a potential nasal absorption enhancer for the nasal delivery of ketoconazole.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.2
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• pp.71-75
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• 2010

To investigate the intestinal absorption of a fibrinolytic and proteolytic lumbrokinase extracted from Eisenia andrei, we used rat everted gut sacs and an in situ closed-loop recirculation method. We extracted lumbrokinase from Eisenia andrei, and then raised polyclonal antibody against lumbrokinase. Fibrinolytic activity and proteolytic activity in the serosal side of rat everted gut sacs incubated with lumbrokinase showed dose- and time-dependent patterns. Immunological results obtained by western blotting serosal side solution using rat everted gut sacs method showed that lumbrokinase proteins between 33.6 and 54.7 kDa are absorbed mostly by the intestinal epithelium. Furthermore, MALDI- TOF mass spectrometric analysis of plasma fractions obtained by in situ recirculation method confirmed that lumbrokinase F1 is absorbed into blood. These results support the notion that lumbrokinase can be absorbed from mucosal lumen into blood by oral administration.

• Toxicological Research
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• v.5 no.1
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• pp.9-15
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• 1989

The effect of butylated hydroxytoluene (BHT) and its major metabolite, 3, 5-di-tert-butyl-4-hydroxybenzoic acid (BHT-acid) on the uptake of taurocholate into hepatocytes was studied using the primary culture of rat hepatocytes. Hepatocyte were isolated by an in situ collagenase perfusion technique and maintained as a monolayer in serum-free meadia for 24 hours before use. The uptake of taurocholate was saturable with an apparent Km of 12.8+2.8 MuM and Vmax of 0.18+0.01 nmol/mg/min. Both BHT and BHT-acid inhibited the hepatocellular uptake of taurocholate when they were added to the culture.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.119-119
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• 1997

Valacyclovir is a L-valyl ester prodrug of acyclovir which is a highly effective and selective antiviral agent in the treatment of herpes virus diseases. Valacyclovir is rapidly and almost completely converted to acyclovir and increases the oral bioavailability of acyclovir three to five fold. However, the intestinal absorption mechanism of valacyclovir is not clear. If the improved absorption mechanism of valacyclovir is fully understood, it will provide a rationale of designing the amino acid ester prodrugs of polar drugs containing hydroxyl group. The main objective of our present study is to characterize the membrane transport mechanism of valacyclovir. Methods : Intestinal absorption of valacyclovir was investigated by using in-situ rat perfusion study and its wall permeability was estimated by modified boundary layer model. The membrane transport mechanism was also investigated through the uptake study in Caco-2 cells and in CHO-hPepTl cells. Results : In the rat perfusion study, the wall permeability of valacyclovir was ten times higher than acyclovir and showed concentration dependency, Valacyclovir also demonstrated a D,L stereo-selectivity with L-isomer having an approximately five-fold higher permeability than D-isomer. Mixed dipeptides and cephalexin, which are transported by dipeptide carriers, strongly competed with valacyclovir for the intestinal absorption, while L-valine did not show any competition with valacyclovir. This indicated that the intestinal absorption of valacyclovir could be dipeptide carrier-mediated. In addition, the competitive uptake study in Caco-2 cells presented that dipeptides reduced the valacyclovir uptake but valine did not. Also, in IC$\sub$50/ study, valacyclovir showed strong inhibition on the $^3$H-gly-sar uptake in CHO-hPepTl cells over-expressing a human intestinal peptide transporter. Taken together, the result from our present study indicated that valacyclovir utilized the peptide transporter for the intestinal absorption.

• Journal of Life Science
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• v.20 no.2
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• pp.169-175
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• 2010

Intestinally absorptive and distributive aspects of the subtoxic level of selenite in rats were investigated using a double perfusion system. The double-perfusion technique is an in situ, in vitro preparation in which the intestinal lumen and its vasculature are perfused simultaneously. In the previous study, the subtoxic level of sodium selenite was determined to be 1.2 mM through inhibition of 3-0-methyl glucose (3MG) absorption. Thus, the selenite used to identify the intestinally absorptive mechanism of selenite was perfused at a luminal concentration of 1, 10, 50, 100 and $200\;{\mu}M$. Appearance of radiolabeled-Selenium (Se) was identified in three compartments: luminal perfusate, small intestine and vascular perfusate. Dose-response curves for Se in the three compartments indicate that selenite is absorbed by non-mediated passive diffusion. Regarding the distributive aspect, $21.02{\pm}3.92%$ of the total amount of selenite in the lumen was transported into the blood vessels across the small intestine. However, $4.75{\pm}1.75%$ of the total amount of selenite in the lumen is retained by the small intestine. Therefore, a total of $25.67{\pm}4.46%$ of the test dose was taken up from the luminal perfusate.