• Journal of Microbiology and Biotechnology
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• 제5권6호
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• pp.315-318
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• 1995

Sulfanilic acid (4-aminobenzenesulfonic acid) alone is normally not permeant in bacteria but can be readily delivered via the microbial dipeptide transport system. A dipeptidyl derivative of this compound, L-phenylalanyl-L-2-sulfanilylglycine (PSG), prepared by attachment of its primary amino group to the phenylalanyl $\alpha$-glycine moiety, appeared to have a Km of 0.125 mM and a Vmax of 1.9 nmoles/ml/min ($A_{660}$, 1.0) in Escherichia coli. From competitive spectrophotometric analysis, it was found that the type of amino acids in both of the N- and C-terminals affected the kinetic power of dipeptides. The growth inhibitory effect of PSG was over 7 times more potent than that of the sulfanilic acid against E. coli, suggesting that this potential inhibition was presumably due to the increased hydrophobic nature of the sulfanilyl dipeptide.

• Journal of Microbiology and Biotechnology
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• 제6권2호
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• pp.92-97
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• 1996

Portage kinetic constants of peptide transport can be measured by competitive spectrophotometry. The kinetic constants of L-Glu-L-Glu transport in Escherichia coli were ascertained using L-Phe-L-3-thia-Phe (PSP) as a detector. Since the production of thiophenol upon intracellular hydrolysis of PSP was competitively inhibited by L-Glu-L-Glu, it was able to compute the kinetic constants of L-Glu-L-Glu using this method. The resulted data were in agreement with the values obtained by the method of Michaelis-Menten kinetics. The potential of this method was examined against dipeptide transport systems in various microorganisms. These results strongly suggest that the overall properties of individual systems for dipeptide transports can be easily characterized by competitive spectrophotometry.

• Asian-Australasian Journal of Animal Sciences
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• 제12권6호
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• pp.939-943
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• 1999

Three 28 day-old $Duroc{\times}Large$ $White{\times}Landrace$ litter mate gilts weighing an average of 6.5 kg were used to study the intestinal absorption of amino acids when provided in dipeptide form or in the form of a free amino acid mixture. The pigs were given one of three treatments. The control involved a duodenal infusion containing no amino-acids (phosphate buffer plus 5% sorbitol) while the remaining two treatments involved either a duodenal infusion containing a glycine-lysine dipeptide (1 g) or a mixture of the free amino acids glycine and lysine at the same concentration as in the dipeptide. Blood was drawn from a cannula inserted in the portal vein, at 5 to 20 minute intervals, for two hours following infusion. The concentration of intact dipeptide as well as free glycine and lysine in the portal blood was determined by high performance liquid chromatography. The intact dipeptide was never detected in the portal blood at any time after infusion. Lysine appeared in the portal blood more rapidly after infusion of dipeptide than after infusion of free lysine and the concentration of lysine in portal blood was higher in the pig infused with the dipeptide than after infusion of free lysine at almost all time points measured. The cumulative absorption of lysine and glycine from the intestine during the two hour period after infusion was greater in the pig infused with dipeptide than in the pig infused with free amino acids. The results suggest that although intact dipeptide did not reach he portal circulation, a special transport mechanism for absorption of dipeptide by intestinal cells appears to be present in pigs similar to that observed in other species.

• Journal of Pharmaceutical Investigation
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• 제25권2호
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• pp.137-143
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• 1995

The peptide transporter can be utilized for improving the bioavailability of compounds that are poorly absorbed. Characterization of the dipeptide uptake into the human intestinal epithelial cells, HT-29 was investigated. The uptake of tritiated glycylsarcosine $([^3H]-Gly-Sar,\;0.1\;{\mu}Ci/ml)$ was measured in confluent or subconfluent HT-29, Caco-2, and Cos-7 cells. Uptake medium was the Dulbecco's Modified Eagle's Media (DMEM) adjusted to pH 6.0. Both HT-29 and Caco-2 cells expressed the dipeptide transporter significantly (p<0.005) but Cos-7 did not. Certain portions of passive uptake were observed in all three cell lines. Uptake of Gly-Sar was largest at 7 days after plating HT-29 cells with significant inhibition with 25 mM cold Gly-Sar (p<0.05). but expression ratio of the dipeptide transporter was 0.7, suggesting lower expression. The effect of pH on Gly-Sar uptake was not significant in the range of pH 6 to 8. Gly-Sar uptake was also inhibited with 50 mM carnosine, 25 mM Gly-Sar, and 35 mM cephalexin significantly (p<0.05). From above results the dipeptide transporter was expressed well in HT-29 cells and was similar to that in the small intestine, suggesting that large amounts of mRNA of the transporter from the cells can be obtained.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1997년도 춘계학술대회
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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.

• Bulletin of the Korean Chemical Society
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• 제14권6호
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• pp.674-678
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• 1993

Di-and tri-peptides containing DL-N-hydroxyalanine were prepared. DL-N-Hydroxyalanine was linked, via its primary amino group, to the ${\alpha}$-carbon of glycine residues in dipeptide synthon (L-alanyl(${\alpha}$-DL-N-hydroxyalanyl)DL-glycine) 5, and tripeptide synthon (L-alanyl-L-alanyl(${\alpha}$-DL-N-hydroxyalanyl) DL-glycine) 12. 5 proved to be 19 times more potent than DL-N-hydroxyalanine when tested in vitro for the ability to inhibit the growth of E coli. However, 12 gave comparable potency to DL-N-hydroxyalanine itself.

• Journal of Pharmaceutical Investigation
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• 제28권1호
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• pp.25-33
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• 1998

Intestinal absorption of ${\beta}-lactam$ antibiotics and angiotensin converting enzyme(ACE) inhibitors has been shown to use the carrier-mediated transport system. In vitro experiments have established that the efficacy of uptake by enterocytes depends on an inwardly directed proton gradient. It was suggested that benazepril was mediated by tripeptide transport system and that amoxicillin was transported by dipeptide transport carrier. The aim of this study is to assess the influence of amoxicillin on the intestinal absorption of benazepril using in vitro diffusion chamber and in situ single pass perfusion technique in the rat in order to elucidate whether the above transport systems are competitive or not. We obtained the gastrointestinal pemeability coefficient of amoxicillin, benazepril and both of them using in vitro diffusion chamber. And also the gastrointestinal absorption clearance of amoxicillin, benazepril and both of them using in situ single-pass perfusion method at steady state were calculated. Amoxicillin and benazepril were analyzed by HPLC. The results by the use of diffusion chamber in vitro indicated that the apparent intestinal permeability coefficient of benazepril was significantly(p<0.01) decreased by amoxicillin(45.2%) and vice versa significantly(p<0.01) decreased(89.1%). The results by the in situ gastrointestinal single-pass perfusion method indicated that the intestinal absorption clearance of benazepril was significantly(p<0.05) decreased by amoxicillin (40.2%) and vice versa significantly(p<0.05) decreased(54.8%). These results might suggest that they share the same peptide carrier pathway for oral absorption.

• Journal of Pharmaceutical Investigation
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• 제27권1호
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• pp.57-64
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• 1997

$Amino-{\beta}-lactam$ antibiotics are absorbed by the dipeptide transporter in the small intestine. These uptakes are coupled to a proton influx. The inward proton gradient is partly induced by the $Na^+/H^+$ exchanger and calcium ion is involved in control of this antiport. Interaction between ampicillin which is one of the $Amino-{\beta}-lactam$ antibiotics and nifedipine which is one of calcium channel blocking agents was studied in rats in vivo and with rabbit jejunum mounted on the Sweetana/Grass diffusion cells in vitro. Bioavailability of ampicillin was increased significantly when nifedipine was co-administered orally in rats. There were no differences in the distribution phase and the elimination phase when ampicillin was given either alone or with nifedipine intravenously. Conditions for in vitro experiments were determined. The lift rate of $O_2/CO_2$ gas was controlled to 3 bubbles/sec and ampicillin was stable in the Kreb's buffer at pH 6.0. Absorption of ampicillin was the greatest when the completely-stripped serosal membrane was used. Transport of ampicillin from mucosal to serosal side in the rabbit jejunum was enhanced by 32% in the presence of nifedipine (p=0.059). Above results suggest that nifedipine might increase the plasma level of ampicillin via the improved absorption in the intestine rather than the reduction in the elimination or/and alteration in the distribution.