Drug Metabolism and Pharmacokinetics
Online ISSN : 1880-0920
Print ISSN : 1347-4367
ISSN-L : 1347-4367
Volume 29 , Issue 3
Showing 1-11 articles out of 11 articles from the selected issue
Editorial
Regular Articles
  • Nan HU, Mei MEI, Jianqing RUAN, Wenjin WU, Yitao WANG, Ru YAN
    2014 Volume 29 Issue 3 Pages 229-236
    Published: June 25, 2014
    Released: June 25, 2014
    [Advance publication] Released: November 19, 2013
    JOURNALS FREE ACCESS
    Oxyresveratrol (OXY) is a natural hydroxystilbene that shows similar bioactivity but better water solubility than resveratrol. This study aims to characterize its glucuronidation kinetics in human liver (HLMs) and intestinal (HIMs) microsomes and identify the main UDP-glucuronosyltransferase (UGT) isoforms involved. Three and four mono-glucuronides of OXY were generated in HIMs and HLMs, respectively, with oxyresveratrol-2-O-β-d-glucuronosyl (G4) as the major metabolite in both organs. The kinetics of G4 formation fit a sigmoidal model in HLMs and biphasic kinetics in HIMs. Multiple UGT isoforms catalyzed G4 formation with the highest activity observed with UGT1A9 followed by UGT1A1. G4 formation by both isoforms followed substrate inhibition kinetics. Propofol (UGT1A9 inhibitor) effectively blocked G4 generation in HLMs (IC50 63.7 ± 11.6 µM), whereas the UGT1A1 inhibitor bilirubin only produced partial inhibition in HLMs and HIMs. These findings shed light on the metabolic mechanism of OXY and arouse awareness of drug interactions.
    Download PDF (1444K)
  • Yuki KONDO, Takahiro IWAO, Katsunori NAKAMURA, Takamitsu SASAKI, Shogo ...
    2014 Volume 29 Issue 3 Pages 237-243
    Published: June 25, 2014
    Released: June 25, 2014
    [Advance publication] Released: December 10, 2013
    JOURNALS FREE ACCESS
    The use of human induced pluripotent stem (iPS) cells would be of great value for a variety of applications involving drug development studies. Several reports have been published on the differentiation of human iPS cells into hepatocyte-like cells; however, the cells were insufficient for application in drug metabolism studies. In this study, we aimed to establish effective methods for differentiation of human iPS cells into hepatocytes. Two human iPS cell lines were differentiated by addition of activin A, dimethyl sulfoxide, hepatocyte growth factor, oncostatin M, and dexamethasone. The differentiated cells expressed hepatocyte markers and drug-metabolizing enzymes, revealing that the human iPS cells were differentiated into hepatocyte-like cells. Expression of CYP3A4 and UGT1A1 mRNAs increased with treatment with typical inducers of the enzymes, and the response of the cells against the inducers was similar to that of human hepatocytes. Furthermore, the drug-metabolizing activity of CYP3A4, as monitored by testosterone 6β-hydroxylase activity, was elevated by these inducers. In conclusion, we established methods for differentiation of hepatocyte-like cells expressing drug metabolizing activity from human iPS cells. The hepatocyte-like cells derived from human iPS cells will be useful for drug metabolism studies.
    Download PDF (1470K)
  • Takahiro KATANO, Yuko ITO, Kinya OHTA, Tomoya YASUJIMA, Katsuhisa INOU ...
    2014 Volume 29 Issue 3 Pages 244-248
    Published: June 25, 2014
    Released: June 25, 2014
    [Advance publication] Released: December 10, 2013
    JOURNALS FREE ACCESS
    Aquaglyceroporins, which constitute a subgroup of aquaporin (AQP) water channels, had been believed to serve as channels for glycerol as well as for water. However, our recent studies have indicated that AQP9 and AQP10 operate in a carrier mode, which is of saturable nature, for glycerol transport. Assuming that such a functional characteristic could also be shared by AQP7, another aquaglyceroporin, we examined its glycerol transport function. The specific transport of glycerol by human AQP7, which was stably expressed in Madin-Darby canine kidney II cells, was indeed highly saturable, indicating the involvement of a carrier mode of operation mechanism. Kinetic analysis indicated that the specific transport conformed to Michaelis-Menten kinetics with the Michaelis constant of 11.9 µM and was not associated with a nonsaturable transport component as an indication of a simultaneous channel mode of operation, which was previously indicated for AQP10. AQP7-specific glycerol transport was furthermore found to be specifically inhibited by several compounds analogous to glycerol and operate without requiring either Na+ or H+. These characteristics of the carrier mode of AQP7 operation suggest that it is a facilitative carrier for glycerol and, possibly, also for analogous compounds, providing a novel insight into its operation mechanism.
    Download PDF (792K)
  • Dawei CHEN, Feng GUO, Jinxiu SHI, Chenhui ZHANG, Zhaowen WANG, Junwei ...
    2014 Volume 29 Issue 3 Pages 249-253
    Published: June 25, 2014
    Released: June 25, 2014
    [Advance publication] Released: December 17, 2013
    JOURNALS FREE ACCESS
    Tacrolimus is a widely used immunosuppressant after organ transplantation. The narrow therapeutic window and individual variability in tacrolimus pharmacokinetics make management of this agent a great challenge. This study was undertaken to determine the association of clinical markers, cytochrome P450, family 3, subfamily A, polypeptide 5 (CYP3A5) and nuclear receptor subfamily 1, group I, member 3 (NR1I3) gene polymorphisms with tacrolimus pharmacokinetics. A total of 96 liver transplant patients were enrolled in the study. Tacrolimus dose-adjusted trough concentration (C/D ratio) and clinical markers were recorded for one month after transplantation. CYP3A5 and NR1I3 gene polymorphisms for both donor and recipient were genotyped. In single variable analysis, hemoglobin (Hb), hematocrit (Hct), donor CYP3A5, NR1I3 gene polymorphisms and recipient CYP3A5 gene polymorphisms were associated with log-transformed tacrolimus C/D ratios. Hb, donor CYP3A5, NR1I3 gene polymorphisms and recipient CYP3A5 gene polymorphisms showed association with log-transformed tacrolimus C/D ratios in the final multiple linear regression model. Donor CYP3A5 polymorphisms were the most important variant, accounting for 14.3% of total variation involved in tacrolimus pharmacokinetics. This information could be useful in developing individualized tacrolimus treatment after liver transplantation.
    Download PDF (131K)
  • Kentaro YANO, Shinobu TAKIMOTO, Toshimitsu MOTEGI, Takumi TOMONO, Miho ...
    2014 Volume 29 Issue 3 Pages 254-258
    Published: June 25, 2014
    Released: June 25, 2014
    [Advance publication] Released: December 24, 2013
    JOURNALS FREE ACCESS
    Cilnidipine is reported to show antihypertensive and neuroprotective actions in a rat brain ischemia model, but is barely distributed to normal brain, suggesting that its uptake into normal brain is inhibited by efflux transporter(s), such as P-glycoprotein (P-gp). Here, we investigated whether P-gp regulates the brain distribution of cilnidipine. Intracellular accumulation of cilnidipine was decreased in P-gp-overexpressing porcine kidney epithelial cells (LLC-GA5-COL150 cells) compared with control LLC-PK1 cells and the decrease was markedly inhibited by verapamil, a P-gp inhibitor. Further, cilnidipine concentration in the brain of P-gp knockout mice was significantly increased after cilnidipine administration, compared with that in wild-type mice. Moreover, when cilnidipine was administered to male spontaneously hypertensive rats (SHR) with tandem occlusion of the distal middle cerebral and ipsilateral common carotid artery, its concentration in the ischemic hemisphere was 1.6-fold higher than that in the contralateral hemisphere. This result was supported by visualization of cilnidipine distribution using matrix-assisted laser desorption/ionization-time of flight/mass spectrometry (MALDI-TOF/MS) imaging. Our results indicated that cilnidipine is normally excluded from the brain by P-gp-mediated efflux transport, but P-gp function is impaired in ischemic brain and consequently cilnidipine is distributed to the ischemic region.
    Download PDF (1004K)
  • Ryoichi FUJIWARA, Tomoo ITOH
    2014 Volume 29 Issue 3 Pages 259-265
    Published: June 25, 2014
    Released: June 25, 2014
    [Advance publication] Released: December 24, 2013
    JOURNALS FREE ACCESS
    UDP-glucuronosyltransferase (UGT) 2B7 is a membrane protein that catalyzes glucuronidation of endogenous and exogenous substrates. Because UGTs are expressed in the endoplasmic reticulum (ER), their substrates and metabolites need to be transported through the ER membrane. However, insight into the mechanism underlying the transport of substrates/metabolites of UGTs through the ER membrane has not been elucidated. Metabolosome is a functional unit of metabolism consisting of multiple metabolism-related proteins. UGTs might form a metabolosome to facilitate the transport of their substrates and/or metabolites through the ER membrane. In the present study, therefore, extensive protein-protein interactions involving UGT2B7 were determined by a shotgun analysis of immunoprecipitate. Our shotgun analysis revealed that 92 proteins were immunoprecipitated with anti-UGT2B7 antibody in human liver microsomes. We further determined that 42 proteins out of the 92 proteins were specifically immunoprecipitated with the anti-UGT2B7 antibody. In addition to UGT2B7, other microsomal enzymes such as UGT1A, CYP3A4, CYP1A2, and a monoamine oxidase, were included in the list of proteins immunoprecipitated with the anti-UGT2B7 antibody, suggesting that these proteins might form a metabolosome to regulate their functions in the liver. Further analyses are needed to elucidate the roles of those proteins in the enzymatic activity of human UGTs.
    Download PDF (2942K)
  • Tomohisa NAKADA, Tomoko KITO, Katsuhisa INOUE, Satohiro MASUDA, Ken-ic ...
    2014 Volume 29 Issue 3 Pages 266-271
    Published: June 25, 2014
    Released: June 25, 2014
    [Advance publication] Released: December 31, 2013
    JOURNALS FREE ACCESS
    Telaprevir-based triple therapy is a highly effective treatment for chronic hepatitis C. However, adverse reactions include reversible and dose-dependent elevation of serum creatinine levels. We speculated that this effect reflects inhibition of the renal organic cation transporters hOCT2, hMATE1, and hMATE2-K by telaprevir or its metabolites (VRT-127394 and VRT-0922061). Telaprevir, VRT-127394, and VRT-0922061 showed negligible or weak effects on hOCT2 at concentrations of ≥20 µM, but inhibited hMATE1 by 35, 38, and 53% and hMATE2-K by 47, 45, and 61% at 100 µM, respectively. Telaprevir or its metabolites (10 µM) did not affect basal-to-apical transport of MPP+ across monolayers of hOCT2-hMATE1 double-transfected MDCKII cells, whereas pyrimethamine, a potent inhibitor of hMATE1, markedly inhibited MPP+ transport. Taken together, inhibition of hOCT2, hMATE1, and hMATE2-K is unlikely to be clinically relevant because unbound plasma concentrations of telaprevir and its metabolites reach only 2 µM following oral administration of a dose of 750 mg telaprevir. Hence, elevated serum creatinine during telaprevir therapy may not be related to direct inhibition of renal organic cation transporters.
    Download PDF (2427K)
  • Koji KIMURA, Risa TAKAYANAGI, Haruko YOKOYAMA, Yasuhiko YAMADA
    2014 Volume 29 Issue 3 Pages 272-277
    Published: June 25, 2014
    Released: June 25, 2014
    [Advance publication] Released: January 14, 2014
    JOURNALS FREE ACCESS
    TNF inhibitors are used as therapeutic agents for rheumatoid arthritis (RA). Each has a different dosage regimen and it is thought that the differences among them have implications on efficacy. However, those differences have not been analyzed in a theoretical manner. In the present study, we tried to explain theoretically the differences. We theoretically analyzed the anti-inflammatory effect of infliximab (IFX), etanercept (ETN), and adalimumab (ADA) for RA by using a pharmacokinetic and pharmacodynamic model. Then, we simulated values for sequential changes of tender joint count (TJC) after repeated administrations of TNF inhibitors by using the model. The sequential changes of TJC obtained with our model were in good agreement with observed TJC ratio data, which was considered to show the validity of our analytical method. The following results were obtained: the onset of clinical response was fastest with IFX, the fluctuation of IFX was greater than that of the others, and the clinical response with ADA was as stable as that with ETN. The present model was useful to analyze theoretically the anti-rheumatic effect of TNF inhibitors. Our results showed that different dosage regimens have implications on the onset and fluctuation of clinical response.
    Download PDF (1348K)
  • Kiyohiko NAKAI, Russell WADA, Satofumi IIDA, Takehiko KAWANISHI, Yoshi ...
    2014 Volume 29 Issue 3 Pages 278-282
    Published: June 25, 2014
    Released: June 25, 2014
    [Advance publication] Released: January 14, 2014
    JOURNALS FREE ACCESS
    Orlistat is used clinically worldwide as anti-obesity drug. It is a chemically synthesized hydrogenated derivative of lipstatin and is an inhibitor of gastric and pancreatic lipases. It has been found to reduce the absorption of dietary fat in the gastrointestinal tract. Modeling and simulation based on pharmacokinetic/pharmacodynamic analysis is becoming increasingly used in the design of clinical trials to assure that the trials are of high quality and are conducted efficiently. We developed a clinical trial simulation model for orlistat based on Phase III clinical study data. This innovative weight loss model includes the relationships between orlistat dose, changes in fecal fat excretion, and weight loss, and also incorporates a dropout function. The model guided the dose-finding strategy and allowed simulation of long-term clinical outcomes of orlistat.
    Download PDF (703K)
Note
  • Tomoko OTA, Mariko HAYASHIDA, Minori ISHII, Kyoko IWAO-KOIZUMI, Shigen ...
    2014 Volume 29 Issue 3 Pages 283-285
    Published: June 25, 2014
    Released: June 25, 2014
    [Advance publication] Released: December 31, 2013
    JOURNALS FREE ACCESS
    In the post-genome era, a simple and inexpensive method for diagnostic analysis is in high demand. Cytochrome P450 (CYP) 2D6 is one of the most widely investigated CYPs in relation to genetic polymorphism. Detection of CYP2D6*5 is difficult since long PCR is used. Especially for samples without DNA extraction, the detection is not sensitive enough for population analysis. Therefore, we developed a CYP2D6*5 genotyping method that involves nested long PCR, directly using human whole saliva as a template without DNA extraction. This method will be very useful for genetic diagnoses and can be an efficient tool for individualization of drug therapy in clinical studies.
    Download PDF (338K)
feedback
Top