Drug Metabolism and Pharmacokinetics
Online ISSN : 1880-0920
Print ISSN : 1347-4367
ISSN-L : 1347-4367
Volume 27 , Issue 3
Showing 1-13 articles out of 13 articles from the selected issue
Editorial
Regular Articles
  • Fumiyoshi YAMASHITA, Atsuto FUJITA, Xingyi ZHANG, Yukako SASA, Kiyoshi ...
    2012 Volume 27 Issue 3 Pages 280-285
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: December 06, 2011
    JOURNALS FREE ACCESS
      Establishment of in vitro–in vivo correlation (IVIVC) accelerates optimization of desirable drug formulations and/or modification of the manufacturing processes in the scale-up and post-approval periods. This article presents a method of finding the optimal conversion function for establishing Level A point-to-point IVIVC, based on a computer-based evolutionary search technique. Gene expression programming (GEP) is a technique for optimizing a mathematical expression tree with the help of a genetic algorithm. A parameter optimization routine, which minimizes the number of parameters in the mathematical expression trees and estimates the best-fit parameter values, was implemented in the GEP algorithm. Feasibility of the computer program was investigated using the in vitro and in vivo data for sustained release diltiazem formulations. It provided a mathematical equation that, from their in vitro dissolution profiles, successfully predicts the plasma concentration profiles of three different formulations of diltiazem following oral administration. Because the present approach does not use intravenous injection data like conventional IVIVC analyses, it is widely applicable to the evaluation of various oral formulations.
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  • Tae-Sung KOO, Hyun-Jung KWON, Mi-Hwa KIM, Dae-Duk KIM, Chang-Koo SHIM, ...
    2012 Volume 27 Issue 3 Pages 286-293
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: December 13, 2011
    JOURNALS FREE ACCESS
      The objective of this study was to identify the nitrogen oxide form(s) involved in the functional impairment of the rat taurine transport system. Taurine uptake activity in the rat renal brush border membrane vesicle (RBBMV) preparation or Xenopus laevis oocytes that express the rat taurine transporter was compared after the pretreatment with nitrogen oxide donors from which nitric oxide (NO) is released at different rates. The functional impairment was associated with a reduced Vmax, but did not involve an alteration in the Km, of taurine uptake in the RBBMV preparation that had been pretreated with sodium nitroprusside, a slow release nitric oxide (NO) donor. When the preparation was pretreated with S-nitroso-N-acetyl penicillamine, a rapid release NO donor, the activity of taurine uptake was unaffected. The activity was not statistically different from the control after the pretreatment with sodium nitroprusside and superoxide dismutase. Consistent with the study with RBBMV, a similar alteration in the activity of taurine uptake by NO donors was observed in oocytes expressing the transporter. Considering the fact that peroxynitrite, a highly reactive nitrogen oxide form, is formed by the reaction between NO and superoxide, the taurine transporter, and probably other transport systems as well, may be functionally impaired by peroxynitrite.
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  • Satoshi YAMAORI, Kyoko KOEDA, Mika KUSHIHARA, Yui HADA, Ikuo YAMAMOTO, ...
    2012 Volume 27 Issue 3 Pages 294-300
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: December 13, 2011
    JOURNALS FREE ACCESS
      Inhibitory effects of Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol (CBD), and cannabinol (CBN), the three major constituents in marijuana, and polycyclic aromatic hydrocarbons (PAHs) contained in marijuana smoke on catalytic activity of human cytochrome P450 (CYP) 2C9 were investigated. These phytocannabinoids concentration-dependently inhibited S-warfarin 7-hydroxylase and diclofenac 4′-hydroxylase activities of human liver microsomes (HLMs) and recombinant CYP2C9 (rCYP2C9). In contrast, none of the twelve PAHs including benz[a]anthracene and benzo[a]pyrene exerted substantial inhibition (IC50 > 10 µM). The inhibitory potentials of Δ9-THC (Ki = 0.937–1.50 µM) and CBN (Ki = 0.882–1.29 µM) were almost equivalent regardless of the enzyme sources used, whereas the inhibitory potency of CBD (Ki = 0.954–9.88 µM) varied depending on the enzyme sources and substrates used. Δ9-THC inhibited both S-warfarin 7-hydroxylase and diclofenac 4′-hydroxylase activities of HLMs and rCYP2C9 in a mixed manner. CBD and CBN competitively inhibited the activities of HLMs and rCYP2C9, with the only notable difference being that CBD and CBN exhibited mixed-type inhibitions against diclofenac 4′-hydroxylation and S-warfarin 7-hydroxylation, respectively, by rCYP2C9. None of Δ9-THC, CBD, and CBN exerted metabolism-dependent inhibition. These results indicated that the three major phytocannabinoids but not PAHs contained in marijuana smoke potently inhibited CYP2C9 activity and that these cannabinoids can be characterized as direct inhibitors for CYP2C9.
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  • Kazuhiko HANADA, Kenya NAKAI, Hiromasa TANAKA, Fumitaka SUZUKI, Hiromi ...
    2012 Volume 27 Issue 3 Pages 301-306
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: December 13, 2011
    JOURNALS FREE ACCESS
      Analysis of mRNAs from liver biopsy samples of patients with chronic hepatitis C revealed that the levels of nuclear receptor expression were correlated with those of drug-metabolizing enzymes and transporters in relation to the development of fibrosis. Overall, the median mRNA level was largely dependent on fibrosis stage (F), and that for stage 3 patients (F3) was about 50% less than that for F1 patients. Levels of expression of AhR, together with CAR and PXR, were lowest in livers of F3 patients. Multivariate linear regression analysis revealed that AhR expression appeared to be involved in the regulation of CYP1A2, 2E1, 2D6, UGT1A, MDR1/3, MRP2/3, NTCP and OCT1 in the livers of patients with chronic hepatitis C. These results suggest that downregulation of AhR during the progression of liver fibrosis is associated with decreased expression levels of these phase I and II enzymes and drug transporters during inflammation-related signal transduction between AhR and other nuclear receptors.
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  • Ryota ISE, Yasuharu NAKANISHI, Sakae KOHARA, Hiroyuki YAMASHITA, Tsuyo ...
    2012 Volume 27 Issue 3 Pages 307-316
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: December 13, 2011
    JOURNALS FREE ACCESS
      Cynomolgus macaques, frequently used in drug metabolism studies, are bred mainly in the countries of Asia; however, comparative studies of drug metabolism between cynomolgus macaques bred in these countries have not been conducted. In this study, hepatic gene expression profiles of cynomolgus macaques bred in Cambodia (mfCAM), China (mfCHN), and Indonesia (mfIDN) were analyzed. Microarray analysis revealed that expression of most hepatic genes, including drug-metabolizing enzyme genes, was not substantially different between mfCAM, mfCHN, and mfIDN; only 1.1% and 3.0% of all the gene probes detected differential expression (>2.5-fold) in mfCAM compared with mfCHN and mfIDN, respectively. Quantitative polymerase chain reaction showed that the expression levels of 14 cytochromes P450 (P450s) important for drug metabolism did not differ (>2.5-fold) in mfCAM, mfCHN, and mfIDN, validating the microarray data. In contrast, expression of CYP2B6 and CYP3A4 differed (>2.5-fold, p < 0.05) between cynomolgus (mfCAM, mfCHN, or mfIDN) and rhesus macaques, indicating greater differences in expression of P450 genes between the two lineages. Moreover, metabolic activities measured using 14 P450 substrates did not differ substantially (<1.5-fold) between mfCAM and mfCHN. These results suggest that gene expression profiles, including drug-metabolizing enzyme genes such as P450 genes, are similar in mfCAM, mfCHN, and mfIDN.
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  • Ryo MATSUYAMA, Masatoshi TOMI, Shin-ichi AKANUMA, Akiko TABUCHI, Yoshi ...
    2012 Volume 27 Issue 3 Pages 317-324
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: December 20, 2011
    JOURNALS FREE ACCESS
      We investigated the regulation of L-type amino acid transporter 1 (LAT1) in a conditionally immortalized rat retinal capillary endothelial cell line (TR-iBRB2 cells) in response to glucose deprivation. The amounts of LAT1 and 4F2 heavy chain (4F2hc) mRNA in TR-iBRB2 cells exposed to glucose-free culture medium for 8 to 24 h were significantly elevated compared with those in control medium. Concomitantly, [3H]L-leucine uptake activity was increased, suggesting that LAT1 transport activity is induced under glucose-deprivation. To determine the transcriptional activity of the LAT1 gene under glucose-free conditions, the promoter activity of the LAT1 gene of approximately 2 kbp (−1958 bp to +70 bp) in TR-iBRB2 cells was assayed using a dual-luciferase reporter assay system. The transcriptional activity of the 2 kbp LAT1 promoter under the glucose-free conditions was 1.7-fold greater than that under normal glucose conditions. The presence of an activator site(s) between −162 bp and −155 bp was indicated by the low activities exhibited by the construct spanning this region and mutagenesis. These results suggest that the glucose deprivation sensitivity of LAT1 expression is transcriptionally regulated, and cis-elements within the LAT1 promoter region from −162 bp to −155 bp mediate this regulation.
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  • Yuko AKIYAMA, Ken-ichi FUJITA, Hiroo ISHIDA, Yu SUNAKAWA, Keishi YAMAS ...
    2012 Volume 27 Issue 3 Pages 325-335
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: December 27, 2011
    JOURNALS FREE ACCESS
      This exploratory retrospective study examined the effects of polymorphisms in transporter genes related to irinotecan pharmacokinetics and those in genes related to irinotecan pharmacodynamics on the efficacy of first-line combination chemotherapy with irinotecan, 5-fluorouracil, and folinic acid (leucovorin) (FOLFIRI) in Japanese patients with advanced colorectal cancer. All patients harbored UDP-glucuronosyltransferase (UGT) 1A1*1/*1, *1/*6, or *1/*28 genotypes, which are associated with similar irinotecan pharmacokinetics and responses to FOLFIRI. Genetic polymorphisms were analyzed by direct sequencing. Overall response rate and median progression-free survival in a total of 61 patients were 43% and 7.5 months, respectively. The overall response rate was higher in patients with the CC genotype at −24 in ATP-binding cassette, subfamily C, and member 2 (ABCC2) than in the others (p = 0.0313). Median progression-free survival was the longest in patients with CC at −24 in ABCC2, followed by those with CT and TT (p = 0.00910). A clear gene-dose effect was seen between −24C>T and median progression-free survival. No other polymorphisms tested were related to the efficacy of FOLFIRI. We thus found that the −24C>T polymorphism in the ABCC2 gene was significantly associated with the efficacy of first-line FOLFIRI in Japanese patients with advanced colorectal cancer.
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  • Ryoko YUMOTO, Sayuri SUZUKA, Keisuke ODA, Junya NAGAI, Mikihisa TAKANO
    2012 Volume 27 Issue 3 Pages 336-343
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: January 03, 2012
    JOURNALS FREE ACCESS
      The uptake mechanism of FITC-labeled albumin (FITC-albumin) was examined in human alveolar epithelial cell line A549. FITC-albumin uptake by A549 cells was time- and temperature-dependent, and was markedly suppressed at 4°C compared with that at 37°C. The uptake was saturable, and was mediated by a high-affinity, low-capacity system and by a low-affinity, high-capacity system. In the following experiments, we focused on the low-affinity system. FITC-albumin uptake was markedly inhibited by metabolic inhibitors and by a vacuolar H+-ATPase, bafilomycin A1. The uptake was inhibited by clathrin-mediated endocytosis inhibitors (phenylarsine oxide and chlorpromazine). Potassium depletion and hypertonicity that inhibit clathrin-mediated endocytosis also decreased FITC-albumin uptake. On the other hand, caveolae-mediated endocytosis inhibitors (indomethacin and nystatin) did not affect FITC-albumin uptake. In addition, FITC-albumin uptake was inhibited by macropinocytosis inhibitors such as 5-(N-ethyl-N-isopropyl) amiloride. These results suggest that the low-affinity system of FITC-albumin uptake is mediated by endocytosis in A549 cells, predominantly via a clathrin-mediated pathway. Macropinocytosis, but not caveolae-mediated endocytosis, may also be involved. Considering our previous findings, albumin may be transported by a similar mechanism and/or pathway in rat and human alveolar epithelial cells.
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Notes
  • Shotaro UEHARA, Norie MURAYAMA, Hiroshi YAMAZAKI, Yasuhiro UNO
    2012 Volume 27 Issue 3 Pages 344-348
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: November 29, 2011
    JOURNALS FREE ACCESS
      Cynomolgus CYP2C76, not orthologous to any human cytochrome P450, partly accounts for species differences in drug metabolism between cynomolgus macaques and humans. To discover the CYP2C76 variants, we previously surveyed cynomolgus macaque genomes and found several non-synonymous variants, including a null allele. However, the analysis was limited to cynomolgus macaques, and the number of genomes was relatively small. In this study, therefore, further screening was conducted using 74 cynomolgus and 30 rhesus macaques. A total of 18 non-synonymous variants was found, among which 7 were in substrate recognition sites, important for protein function, and 14 (74%) were shared by both macaque lineages. In cynomolgus macaques, 3 (16%) non-synonymous variants were unique to Indochinese animals, whereas all the variants found in Indonesian animals were shared by Indochinese animals. Among the 18 variants, as compared with the wild type, in progesterone 16α-hydroxylation, L65F, M310L, and N364S variants showed lower metabolic activity and lower intrinsic clearance by kinetic analysis. Molecular modeling indicated that the reduced catalytic activity of the L65F variant in progesterone 16α-hydroxylation possibly resulted from a longer distance of progesterone to the heme in the active site of the CYP2C76 protein. L65F, M310L, and N364S variants might partly influence inter-animal variations of CYP2C76 metabolic activities.
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  • Erzsébet BEÉRY, Zsuzsanna RAJNAI, Tibor ABONYI, Ildik&oa ...
    2012 Volume 27 Issue 3 Pages 349-353
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: November 29, 2011
    JOURNALS FREE ACCESS
      We are showing that chlorothiazide, a diuretic, is an ABCG2 substrate. It is a Biopharmaceutics Classification System/Biopharmaceutics Drug Distribution and Classification System (BCS/BDDCS) Class IV drug with low bioavailability. Therefore, we tested if chlorothiazide interacts with major apically located intestinal efflux transporters. Our data show that chlorothiazide is transported by ABCG2 with a Km value of 334.6 µM and does not interact with ABCB1 or ABCC2. The chlorothiazide–ABCG2 interaction results in a vectorial transport in MDCKII-BCRP and Caco-2 cells with efflux ratios of 36 and 8.1 respectively. Inhibition of ABCG2 in Caco-2 cells reduced the efflux ratio to 1.4, suggesting that ABCG2 plays a role in limiting chlorothiazide bioavailability in humans.
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Short Communications
  • Masataka OITATE, Shintaro NAKAYAMA, Takashi ITO, Atsushi KURIHARA, Nor ...
    2012 Volume 27 Issue 3 Pages 354-359
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: December 06, 2011
    JOURNALS FREE ACCESS
      We have previously reported that human total body clearance (CL) and steady-state volume of distribution (Vss) of monoclonal antibodies (mAbs) could be predicted reasonably well from monkey data alone using simple allometry with scaling exponents of 0.79 and 1.12 (for soluble targets), and 0.96 and 1.00 (for membrane-bound targets). In the present study, to predict the plasma concentration-time profiles of mAbs in humans, we employed simple dose-normalization and species-invariant time methods (elementary Dedrick plot and complex Dedrick plot), based on the monkey data and the scaling exponents we previously determined. The results demonstrated that the species-invariant time methods were able to provide higher accuracy of prediction than simple dose-normalization, regardless of the type of target antigens (soluble or membrane-bound). The accuracy between elementary Dedrick plot and complex Dedrick plot was nearly equivalent. The predicted human CL and Vss using species-invariant time methods were within mostly 2-fold differences from the observed values. The prediction not only of pharmacokinetic (PK) parameters but also of the plasma concentration-time profile in humans can serve as guidelines for better planning of clinical studies on mAbs.
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  • Yoshiyuki SHIRASAKA, Takanori MORI, Megumi SHICHIRI, Takeo NAKANISHI, ...
    2012 Volume 27 Issue 3 Pages 360-364
    Published: 2012
    Released: June 25, 2012
    [Advance publication] Released: December 27, 2011
    JOURNALS FREE ACCESS
      The purpose of this study was to examine whether the presence of multiple binding sites can explain the pleiotropy of substrate recognition by OATP2B1, using Xenopus oocytes expressing OATP2B1. OATP2B1-mediated uptake of estrone-3-sulfate apparently exhibited biphasic saturation kinetics, with Km values of 0.10 ± 0.05 and 29.9 ± 12.1 µM and Vmax values of 14.1 ± 6.4 and 995 ± 273 fmol/min/oocyte for high- and low-affinity sites, respectively. Contribution analysis revealed that transport of estrone-3-sulfate mediated by high- and low-affinity sites on OATP2B1 could be evaluated at the concentrations of 0.005 and 50 µM, respectively. pH-dependence study of OATP2B1-mediated estrone-3-sulfate uptake suggested that high- and low-affinity sites show different pH sensitivity. When the inhibitory effect of 12 compounds on estrone-3-sulfate uptake by high- and low-affinity sites on OATP2B1 was examined, 4 compounds appeared to be inhibitors of the high-affinity site on OATP2B1. Two other compounds appeared to be inhibitors for the low-affinity site and four others were inhibitory at both sites. These results indicated the presence of multiple binding sites on OATP2B1 with different affinity for drugs. Accordingly, it is likely that drug-drug and drug-beverage interactions occur only when two drugs share the same binding site on OATP2B1.
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