Drug Metabolism and Pharmacokinetics
Online ISSN : 1880-0920
Print ISSN : 1347-4367
ISSN-L : 1347-4367
Volume 23 , Issue 5
Showing 1-12 articles out of 12 articles from the selected issue
Editorial
Regular Articles
  • Katsutoshi HARA, Kazuaki SUYAMA, Hiroshi ITOH, Satoru NAGASHIMA
    2008 Volume 23 Issue 5 Pages 306-312
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      This study was performed to investigate the pharmacokinetics of valaciclovir (VACV), aciclovir (ACV) and 9-(carboxymethoxy)methylguanine (CMMG) in Japanese chronic hemodialysis patients following a single oral administration of 1000 mg VACV and the influence of genetic polymorphisms of aldehyde dehydrogenase-2 (ALDH2) on their pharmacokinetics. A total of eighteen individuals genotyped as ALDH2*1/*1, ALDH2*1/*2 or ALDH2*2/*2 were enrolled in this study. Blood samples were obtained pre-dose and up to 48 hour post-dose. ACV t1/2 was significantly affected by ALDH2 genotype and prolonged in the order of ALDH2*1/*1 (18.1 hr)<ALDH2*1/*2 (21.9 hr)<ALDH2*2/*2 (26.7 hr). ACV AUC from zero to infinity (AUC0-∞) increased with prolonged ACV t1/2. ACV Cmax was similar across the three ALDH2 genotype groups. There was no apparent relationship between ALDH2 genotype and VACV or CMMG pharmacokinetics. This is the first study to show an association between ALDH2 genetic polymorphisms and ACV elimination rate (t1/2) in Japanese end-stage renal disease patients.
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  • Masahiro SHIMOMURA, Satohiro MASUDA, Maki GOTO, Toshiya KATSURA, Tetsu ...
    2008 Volume 23 Issue 5 Pages 313-317
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      The profiles of tacrolimus metabolites in the whole blood and bile were examined in two living-donor liver transplant patients, who transiently required higher doses of tacrolimus. Even when the 16 mg/day or oral 10 mg/day and intravenous infusion of 0.5 mg/day of tacrolimus were administered, its trough level in each patient did not reach over 15 ng/mL. By use of liquid chromatography-tandem mass spectrometry/mass spectrometry methods, a minor metabolite M-II was found to be a major metabolite both in blood and bile in these cases. However, a primary metabolite M-I was confirmed as the majority in the bile of other 8 control cases. Each graft liver and native intestine carried CYP3A5*1/*3 or *3/*3 and *1/*3 or *1/*3, respectively. Therefore, the CYP3A5 genotype could not explain the present phenomena. After removing the bile drainage tube to allow the bile flow into intestine, the required doses of tacrolimus were decreased to around 20% compared to each maximum dosage. In conclusion, a minor metabolite M-II was first found in the human bile, suggesting that the appearance of M-II in bile could associate with the extensive metabolism of tacrolimus and/or the requirement of larger oral dosage.
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  • Maki TAGAWA, Ryoko YUMOTO, Keisuke ODA, Junya NAGAI, Mikihisa TAKANO
    2008 Volume 23 Issue 5 Pages 318-327
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      FITC-albumin uptake by cultured alveolar type II epithelial cells, RLE-6TN, is mediated by high- and low-affinity transport systems.1) In this study, characteristics of the low-affinity transport system were evaluated. The uptake of FITC-albumin was time and temperature dependent and was inhibited by metabolic inhibitors and bafilomycin A1. Confocal laser scanning microscopic analysis showed punctate localization of the fluorescence in the cells, which was partly localized in lysosomes. FITC-albumin taken up by the cells gradually degraded over time, as shown by fluoroimage analyzer after SDS-PAGE. The uptake of FITC-albumin by RLE-6TN cells was not inhibited by caveolae-mediated endocytosis inhibitors such as nystatin, but was inhibited by clathrin-mediated endocytosis inhibitors such as phenylarsine oxide. The uptake was also inhibited by potassium depletion and hypertonicity, conditions known to inhibit clathrin-mediated endocytosis. In addition, macropinocytosis inhibitors such as 5-(N-ethyl-N-isopropyl) amiloride inhibited the uptake. These results indicate that the low-affinity transport of FITC-albumin in RLE-6TN cells is at least in part mediated by clathrin-mediated endocytosis, but not by caveolae-mediated endocytosis. Possible involvement of macropinocytosis was also suggested.
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  • Yoshihiro SHIBATA, Hiroyuki TAKAHASHI, Masato CHIBA, Yasuyuki ISHII
    2008 Volume 23 Issue 5 Pages 328-339
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      A novel method for the prediction of drug-drug interaction has been established based on the in vitro metabolic stability in the “serum incubation method” using cryopreserved human hepatocytes suspended in 100% human serum. As a novel approach, the inhibitory effect of inhibitors on the metabolism of substrates during the first-pass elimination process in the liver (hepatic availability) and in the elimination process from the systemic circulation (hepatic clearance) were separately predicted with the anticipated inhibitor/substrate concentrations during absorption and in the systemic circulation, respectively. Ketoconazole strongly inhibited CYP3A4-mediated terfenadine metabolism in vitro, and the method predicted 6- to 37-fold increase of terfenadine AUC by the concomitant dosing of ketoconazole, which reasonably well agreed with the observed 13- to 59-fold increase of AUC in clinical studies. The CYP3A4-mediated metabolism of indinavir was also subject to the inhibition by ketoconazole in vitro at the lower indinavir concentration (2 μM), whereas no substantial inhibition was observed at 12 μM due to the saturation of indinavir metabolism. Predicted no interaction between ketoconazole and indinavir was consistent with the minimal increase (1.3-fold increase) of indinavir AUC by ketoconazole observed in clinical study. In addition, the method was applied to the CYP2D6-mediated desipramine-quinidine interaction: the predicted 6.4-fold increase of desipramine AUC by quinidine was consistent with the observed 6.7-fold increase of AUC in the clinical drug-drug interaction study. On the other hand, desipramine metabolism was little affected by ketoconazole in vitro, and consequently, it predicted no drug-drug interaction between desipramine and ketoconazole in humans, which agreed with the negligible interaction observed in clinical study. The accuracy of predictions for drug-drug interaction by the serum incubation method was evaluated by comparing the predicted increase of AUC after an oral administration by the inhibitor with the corresponding drug-drug interaction reported from clinical studies. These data demonstrated that the newly established method provides an in vitro tool for the prediction of drug-drug interaction with the accuracy ranging from 0.46 to 1.5.
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  • Katsutoshi TAHARA, Yuka KAGAWA, Mari TAKAAI, Masato TAGUCHI, Yukiya HA ...
    2008 Volume 23 Issue 5 Pages 340-346
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      To evaluate the mechanism responsible for the tubular secretion of bisoprolol, we compared transcellular transport of bisoprolol with that of tetraethylammonium (TEA), cimetidine, and quinidine across LLC-PK1 cell monolayers grown on porous membrane filters. TEA and cimetidine were actively transported in the basolateral-to-apical direction by the specific transport system. Pharmacokinetic analysis indicated that basolateral influx and apical efflux were cooperatively responsible for the directional transport of TEA and cimetidine. Lipophilic cationic drugs, quinidine, S-nicotine, and bisoprolol, significantly diminished basolateral influx and apical efflux clearance of cimetidine. However, transcellular transport of quinidine in the basolateral-to-apical direction was similar to that in the opposite direction in LLC-PK1 cells. In contrast, quinidine was transported actively in the basolateral-to-apical direction in P-glycoprotein-expressed LLC-GA5-COL150 cells. Pharmacokinetic analysis indicated that P-glycoprotein increased the apical efflux of quinidine and also decreased the apical influx of the drug. Basolateral-to-apical transport of bisoprolol was also similar to apical-to-basolateral transport in LLC-PK1 cells, whereas the drug was directionally transported from the basolateral to the apical side in LLC-GA5-COL150 cells. These results suggested that bisoprolol was not significantly transported via transport systems involved in the directional transport of TEA and cimetidine, but that P-glycoprotein was responsible for the directional transport of bisoprolol as well as quinidine in renal epithelial cells.
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  • Masanori NAKAKARIYA, Taiki SHIMADA, Masanori IROKAWA, Tomoji MAEDA, Ik ...
    2008 Volume 23 Issue 5 Pages 347-355
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      The present study was designed to identify the organic anion transporting polypeptide (OATP) molecule(s) responsible for the uptake of β-lactam antibiotics in human liver, using cryopreserved hepatocytes, as well as Xenopus oocytes and cultured cells expressing human OATPs. Nafcillin uptake by human hepatocytes was saturable with a Km of 533 μM. In vitro uptake studies revealed that OATP1B3 and OATP1B1 transported nafcillin with Km values of 74 μM and 11 mM, respectively. Analysis by the relative activity factor method suggested that OATP1B3 contributes mainly to nafcillin uptake and OATP1B1 contributes moderately. This conclusion was supported by the results of a study with selective inhibitors. Furthermore, OATP1B3 transported six other β-lactam antibiotics, and their uptake clearances by OATP1B3 correlated well with those mediated by rat Oatp1a4, which is the predominant contributor to basolateral uptake of nafcillin by rat hepatocytes. These findings suggest that OATP1B3 plays a major role in the hepatic uptake of β-lactam antibiotics in humans, and probably corresponds functionally to Oatp1a4 in rat liver.
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  • Mikio TOMITA, Yusuke TAKIZAWA, Hisanao KISHIMOTO, Masahiro HAYASHI
    2008 Volume 23 Issue 5 Pages 356-363
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      We presented the ischemia/reperfusion (I/R) model which can evaluate changes in P-glycoprotein (P-gp) function induced by lipid peroxidation using surgical-sutures connected with the spring balance. The superior mesenteric artery and vein was occluded by hanging itself using surgical-sutures connected with the spring balance for 60 min (ischemia), followed by reperfusion by cutting of sutures. To determine the hanging force of blood vessel during ischemia, treatment at the hanging force of 50g load, 100g load and 150g load for 60 min was carried out and survival rate was evaluated. Although our 150g load group had some effect on survival, the survival was 100% in the case of 50g and 100g load groups. Thiobarbituric acid-reactive substance (TBA-RS) as an indicator of lipid peroxidation and P-gp expression level after I/R was increased and decreased in a load-dependent manner during ischemia, respectively. Also, the decrease in the level of mdr1a mRNA and function of P-gp by I/R depended on load during ischemia. The changes in TBA-RS, P-gp expression level and P-gp function observed in this study corresponded with our in vitro I/R model reported previously. In conclusion, it was shown that this in vivo I/R model can evaluate the function of P-gp through lipid peroxidation.
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  • Yuan LU, Stacie BRATTON, Jean-Marie HEYDEL, Anna RADOMINSKA-PANDYA
    2008 Volume 23 Issue 5 Pages 364-372
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      Human UDP-glucuronosyltransferase 2B7 (UGT2B7) is one of the major isoforms involved in the glucuronidation of endogenous compounds and xenobiotics. This isoform is the only human UGT shown to glucuronidate retinoids and their oxidized derivatives. In this study, the effects of all-trans retinoic acid (atRA), 9-cis RA, and the RAR agonist TTNPB, on UGT2B7 and UGT2B15 mRNA expression in Caco-2 cells have been examined. Each of these retinoids significantly suppressed UGT2B7 mRNA expression in a concentration-dependent manner with IC50 values of 3.5, 0.3, and 0.2 μM, respectively. However, no inhibition was observed when two other UGTs, UGT2B15 or -1A6, were exposed to atRA, 9-cis RA, or TTNPB, demonstrating that the inhibitory effect of retinoids might be specific for the UGT2B7 isoform. Further, experiments with oxidized atRA derivatives, 4-OH-atRA, 4-oxo-atRA, and 5,6-epoxy-atRA showed that these RA degradation products have no inhibitory effect on UGT2B7 mRNA expression. These data lead us to hypothesize that biologically active forms of RA suppress the expression of UGT2B7 in intestinal cells. This information provides a new pathway by which retinoids may enhance their own toxicity when accumulated in the body at pharmacological concentrations by down-regulating the enzymes involved in their biotransformation into soluble derivatives.
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Note
  • Shiro FUKUMORI, Toshiya MURATA, Mari TAKAAI, Katsutoshi TAHARA, Masato ...
    2008 Volume 23 Issue 5 Pages 373-378
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      The aim of this study was to investigate the involvement of the peptide transporter for absorption of levofloxacin in Caco-2 cells. To evaluate the activity of apical and basolateral peptide transport, we first performed pharmacokinetic analysis of transcellular transport of glycylsarcosine (Gly-Sar) in cell monolayers grown on porous membrane filters. Transcellular transport of Gly-Sar at the medium pH 6 was greater in the apical-to-basolateral direction than in the opposite direction. Influx clearance of Gly-Sar at the apical membrane was much greater than basolateral influx and efflux clearance, indicating that the apical peptide transporter plays an important role in directional transcellular transport of the dipeptide across Caco-2 cell monolayers. We then evaluated the effect of various compounds on the uptake of Gly-Sar and levofloxacin at the apical membrane of Caco-2 cells. The apical uptake of [3H]Gly-Sar was significantly inhibited by Ala-Ala, Gly-Sar, and also levofloxacin, whereas that of [14C]levofloxacin was not inhibited by Ala-Ala and Gly-Sar. On the other hand, the apical uptake of [14C]levofloxacin was inhibited by nicotine, enalapril, fexofenadine, and L-carnitine. These findings indicated that the apical uptake transporter of levofloxacin is distinct from the peptide transporter in Caco-2 cells.
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SNP Communications
  • Su-Ryang KIM, Yoshiro SAITO, Keiko MAEKAWA, Emiko SUGIYAMA, Nahoko KAN ...
    2008 Volume 23 Issue 5 Pages 379-384
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      Deoxycytidine kinase (dCK) is a rate-limiting enzyme in the activation of nucleoside anticancer drugs, such as gemcitabine and cytarabine (Ara-C), to their active metabolites. In this study, the 5′-flanking region, 7 exons and their flanking introns of DCK were comprehensively screened for genetic variations in 256 Japanese cancer patients administered gemcitabine. Twenty-nine genetic variations, including twenty novel ones, were found: 11 in the 5′-flanking region, 1 in the 5′-untranslated region (UTR), 1 in the coding exon, 9 in the 3′-UTR, and 7 in the introns. The novel variations included -1110C>T, -757G>A, -639C>T, -465G>A, -402T>C, -224C>A, -199C>G, IVS1+38G>T, IVS2+78_+83delTTTTTC, IVS3-9C>T, IVS4+12T>C, IVS5+39T>C, 1357A>G, 1545A>T, 1572delA, 1736G>A, 1749G>A, 1838T>C, 1889G>A, and 2048A>T. The frequencies were 0.01 for IVS2+78_ +83delTTTTTC, 0.008 for -402T>C, 0.006 for -639C>T and IVS4+12T>C, 0.004 for -757G>A and 1572delA, and 0.002 for the other 14 variations. A known nonsynonymous SNP 364C>T (Pro122Ser) was detected at a 0.061 frequency. Using the detected polymorphisms, linkage disequilibrium analysis was performed, and 24 haplotypes were identified or inferred. Our findings suggest considerable ethnic differences in genetic variations of DCK and provide fundamental and useful information for genotyping DCK in the Japanese and probably other Asian populations.
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  • Woon Fei HO, Seok Hwee KOO, Jie Yin YEE, Jon Deoon LEE
    2008 Volume 23 Issue 5 Pages 385-391
    Published: 2008
    Released: October 31, 2008
    JOURNALS FREE ACCESS
      MRP2 is a drug transporter that is responsible for the gastrointestinal absorption and biliary excretion of a wide variety of endogenous and xenobiotic compounds, including many clinically used drugs. This study aims to identify genetic variations of ABCC2 gene in three distinct ethnic groups of the Singaporean population (n=288). The coding region of the gene encoding the transporter protein was screened for genetic variations in the study population by denaturing high-performance liquid chromatography and DNA sequencing. Twenty-two genetic variations of ABCC2, including 8 novel ones, were found: 1 in the 5′ untranslated region, 10 in the coding exons (8 nonsynonymous and 2 synonymous variations), and 11 in the introns. Three novel nonsynonymous variations: 2686G>A (Glu896Lys), 4240C>T (His1414Tyr) and 4568A>C (Gln1523Pro) were detected in single heterozygous Malay, Chinese, and Indian subjects, respectively. Among the novel nonsynonymous variations, 4240C>T and 4568A>C were predicted to be functionally significant. These data would provide fundamental and useful information for pharmacogenetic studies on drugs that are substrates of MRP2 in Asians.
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