Drug Metabolism and Pharmacokinetics
Online ISSN : 1880-0920
Print ISSN : 1347-4367
ISSN-L : 1347-4367
Volume 26 , Issue 6
Showing 1-14 articles out of 14 articles from the selected issue
Editorial
Regular Articles
  • Junya HATANAKA, Yukiko SHINME, Kazuki KURIYAMA, Atsushi UCHIDA, Keitat ...
    Type: Regular Article
    2011 Volume 26 Issue 6 Pages 551-558
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: September 13, 2011
    JOURNALS FREE ACCESS
      The main purpose of the present study was to develop a novel formulation of St. John's Wort (SJW) extract with the aim of improving its pharmacokinetics and anti-nociceptive effect. Several formulations of SJW were prepared, including cyclodextrin inclusion (SJW-CD), solid dispersion (SJW-SD), dry-emulsion (SJW-DE), and nano-emulsion (SJW-NE). Physicochemical properties of SJW formulations were characterized with a focus on the morphology, dissolution behavior, colloidal properties, and dispersion stability in water. Although all the SJW formulations and SJW extract itself exhibited fine dissolution behavior in water, SJW extract and most formulations tended to cream, aggregate, or flocculate after dispersion in distilled water. In contrast, there were no significant changes in appearance and particle size of the SJW-NE for at least a few weeks, suggesting that SJW-NE was the most stable form as a carrier of SJW in the present study. After oral administration of the SJW-NE formulation (5.2 mg hyperforin/kg) in mice, higher hyperforin exposure in plasma (1188 ± 41 nM·h) and the brain (52.9 ± 1.6 pmol/g tissue·h) was observed with 2.8- and 1.3-fold increases of the area under the concentration curve from 0 to 6 hours (AUC0–6) compared to those of the SJW extract (417 ± 41 nM·h in plasma and 41.6 ± 1.5 pmol/g tissue·h in the brain). In the formalin test for scoring properties of the first and second phases of the pain response in mice, single oral administration of SJW-NE significantly reduced the nociceptive response compared with SJW extract. From these findings, the NE approach might be efficacious in improving the oral bioavailability and anti-nociceptive effect of SJW extract.
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  • Luigi QUINTIERI, Pietro PALATINI, Stefano MORO, Maura FLOREANI
    Type: Regular Article
    2011 Volume 26 Issue 6 Pages 559-568
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: July 26, 2011
    JOURNALS FREE ACCESS
      The aim of this study was to assess the effects of diosmetin and hesperetin, two flavonoids present in various medicinal products, on CYP2C8 activity of human liver microsomes using paclitaxel oxidation to 6α-hydroxy-paclitaxel as a probe reaction. Diosmetin and hesperetin inhibited 6α-hydroxy-paclitaxel production in a concentration-dependent manner, diosmetin being about 16-fold more potent than hesperetin (mean IC50 values 4.25 ± 0.02 and 68.5 ± 3.3 µM for diosmetin and hesperetin, respectively). Due to the low inhibitory potency of hesperetin, we characterized the mechanism of diosmetin-induced inhibition only. This flavonoid proved to be a reversible, dead-end, full inhibitor of CYP2C8, its mean inhibition constant (Ki) being 3.13 ± 0.11 µM. Kinetic analysis showed that diosmetin caused mixed-type inhibition, since it significantly decreased the Vmax (maximum velocity) and increased the Km value (substrate concentration yielding 50% of Vmax) of the reaction. The results of kinetic analyses were consistent with those of molecular docking simulation, which showed that the putative binding site of diosmetin coincided with the CYP2C8 substrate binding site. The demonstration that diosmetin inhibits CYP2C8 at concentrations similar to those observed after in vivo administration (in the low micromolar range) is of potential clinical relevance, since it may cause pharmacokinetic interactions with co-administered drugs metabolized by this CYP.
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  • Feng LI, Yan MIAO, Lirong ZHANG, Sarah Ann NEUENSWANDER, Justin T. DOU ...
    Type: Regular Article
    2011 Volume 26 Issue 6 Pages 569-576
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: August 16, 2011
    JOURNALS FREE ACCESS
    Supplementary material
      Isoniazid (INH) is a first-line drug for tuberculosis control; the side effects of INH are thought to be associated with its metabolism, and this study was designed to globally characterize isoniazid metabolism. Metabolomic strategies were used to profile isoniazid metabolism in humans. Eight known and seven novel INH metabolites and hydrazones were identified in human urine. The novel products included two hydroxylated INH metabolites and five hydrazones. The two novel metabolites were determined as 2-oxo-1,2-dihydro-pyridine-4-carbohydrazide and isoniazid N-oxide. Five novel hydrazones were produced by condensation of isoniazid with keto acids that are intermediates in the metabolism of essential amino acids, namely, leucine and/or isoleucine, lysine, tyrosine, tryptophan, and phenylalanine. This study enhances our knowledge of isoniazid metabolism and disposition and may offer new avenues for investigating INH-induced toxicity.
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  • Jun TERASHIMA, Wataru HABANO, Toshie GAMOU, Shogo OZAWA
    Type: Regular Article
    2011 Volume 26 Issue 6 Pages 577-583
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: August 30, 2011
    JOURNALS FREE ACCESS
      Cross-talk between the aryl hydrocarbon receptor (AhR) pathway and the typical stress response is thought to be an important signal transduction in response to nutrient-stress conditions, such as glucose deprivation in liver cells. In the present study, we demonstrate that reduction of glucose concentration in the medium of HepG2 cells, a human hepatocellular carcinoma cell line, induces the CYP1 family and Nrf2. RNAi for AhR abolishes the induction of expression of CYP1 and Nrf2. These inductions are accompanied by the translocation of AhR into the nucleus in response to low-glucose conditions. Endogenous compounds are recruited as AhR ligands to induce various gene expressions, and our present results suggest that an endogenous AhR ligand is produced under low-glucose conditions and that the role of AhR as a transcription factor is related to the low-glucose response. The recommended glucose concentration (4.5 g/L) in the medium for culture of HepG2 was used as the high-glucose concentration in this study. We adopted 1.0 g/L as the low-glucose condition for elucidation of mechanisms of the stress response. These results will be useful to understand the relationship between drug-metabolizing enzymes and mechanisms of the anti-stress response of tumor cells, and will also be useful for investigating preventive remedies against tumor angiogenesis.
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  • Ayaka NAWA, Wakako FUJITA-HAMABE, Shiroh KISHIOKA, Shogo TOKUYAMA
    Type: Regular Article
    2011 Volume 26 Issue 6 Pages 584-591
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: August 30, 2011
    JOURNALS FREE ACCESS
      Morphine is one of the strongest analgesics and is commonly used for the treatment of chronic pain. The pharmacokinetic properties of morphine are, in part, modulated by P-glycoprotein (P-gp). We previously reported that intestinal P-gp expression levels are influenced via the activation of inducible nitric oxide synthase (iNOS) in streptozotocin (STZ)-induced diabetic mice. Herein, we examine the analgesic effects of orally administered morphine and its pharmacokinetic properties under diabetic conditions, specifically we focusing on the involvement of intestinal P-gp in a type 1 diabetic mouse model. We assessed the analgesic effect of morphine using the tail-flick test. Serum and brain morphine levels were determined using a HPLC-ECD system. Oral morphine analgesic effects and serum and brain morphine content were significantly increased 9 days after STZ administration. The increase in the analgesic effects of morphine, as well as serum and brain morphine content, was suppressed by aminoguanidine, a specific iNOS inhibitor. Conversely, there were no changes in the analgesic effects obtained with subcutaneous morphine in STZ-treated mice. Our findings suggest that the analgesic effects of oral morphine are dependent on intestinal P-gp expression, and this may be one of the reasons that it is difficult to obtain stable pharmacological effects of morphine in diabetic patients.
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  • Haruka NISHIMUTA, Kimihiko SATO, Masashi YABUKI, Setsuko KOMURO
    Type: Regular Article
    2011 Volume 26 Issue 6 Pages 592-601
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: August 30, 2011
    JOURNALS FREE ACCESS
      This study aimed to establish a practical and simplified method of predicting intestinal availability in humans (Fg,human) at the drug discovery stage using in vitro metabolic clearance values and permeability clearance values. A prediction model for Fg,human of 19 CYP3A substrates and 5 UGT substrates was constructed based on the concept that the permeability clearance values mean the permeability across the basal membrane with a pH of 7.4 on both sides. Permeability clearance values were obtained by parallel artificial membrane permeability assay (PAMPA) at pH 7.4. PAMPA is widely used in the pharmaceutical industry as the earliest primary screening stage and enables estimation of the kinetics of transport by passive diffusion. For CYP3A substrates, the metabolic clearance was obtained from in vitro intrinsic clearance values in human intestinal or hepatic microsomes (CLint,HIM or CLint,HLM, respectively). Using metabolic clearances corrected by the ratio of CLint,HIM to CLint,HLM, HLM showed equivalent predictability to that of HIM for CYP3A substrates. For UGT substrates, the clearance was obtained from alamethicin-activated HIM using one incubation with both NADPH and UDPGA cofactors. The method proposed in this study could predict Fg,human for the compounds investigated and represents a simplified method based on a new concept applicable to lower permeability compounds.
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  • Shin-ichi AKANUMA, Yasuo UCHIDA, Sumio OHTSUKI, Jun-ichi KAMIIE, Masan ...
    Type: Regular Article
    2011 Volume 26 Issue 6 Pages 602-611
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: September 06, 2011
    JOURNALS FREE ACCESS
      β-Lactam antibiotics have cerebral and peripheral adverse effects. Multidrug resistance-associated protein 4 (MRP4) has been reported to transport several β-lactam antibiotics, and its expression at the blood–brain barrier also serves to limit their distribution to the brain. Therefore, the purpose of this study was to clarify the structure–activity relationship of MRP4-mediated transport of β-lactam antibiotics using MRP4-expressing Sf9 membrane vesicles. The transport activity was evaluated as MRP4-mediated transport per MRP4 protein [nL/(min·fmol MRP4 protein)] based on measurement of MRP4 protein expression by means of liquid chromatography-tandem mass spectrometry. Cefotiam showed the greatest MRP4-mediated transport activity [8.90 nL/(min·fmol MRP4 protein)] among the β-lactam antibiotics examined in this study. Measurements of differential transport activity of MRP4 for various β-lactam antibiotics indicated that (i) cephalosporins were transported via MRP4 at a greater rate than were penams, β-lactamase inhibitors, penems, or monobactams; (ii) MRP4-mediated transport activity of anionic cephalosporins was greater than that of zwitterionic cephalosporins; and (iii) higher-molecular-weight anionic β-lactam antibiotics showed greater MRP4-mediated transport activity than lower-molecular-weight ones, whereas zwitterionic β-lactam antibiotics did not show molecular weight dependency of MRP4-mediated transport. These quantitative data should prove useful for understanding MRP-related adverse effects of β-lactam antibiotics and their derivatives.
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  • Azusa HOSHINO-YOSHINO, Motohiro KATO, Kohnosuke NAKANO, Masaki ISHIGAI ...
    Type: Regular Article
    2011 Volume 26 Issue 6 Pages 612-620
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: September 06, 2011
    JOURNALS FREE ACCESS
      The purpose of this study was to provide a pharmacokinetics/pharmacodynamics and toxicokinetics/toxicodynamics bridging of kinase inhibitors by identifying the relationship between their clinical and preclinical (rat, dog, and monkey) data on exposure and efficacy/toxicity. For the eight kinase inhibitors approved in Japan (imatinib, gefitinib, erlotinib, sorafenib, sunitinib, nilotinib, dasatinib, and lapatinib), the human unbound area under the concentration-time curve at steady state (AUCss,u) at the clinical dose correlated well with animal AUCss,u at the no-observed-adverse-effect level (NOAEL) or maximum tolerated dose (MTD). The best correlation was observed for rat AUCss,u at the MTD (p < 0.001). Emax model analysis was performed using the efficacy of each drug in xenograft mice, and the efficacy at the human AUC of the clinical dose was evaluated. The predicted efficacy at the human AUC of the clinical dose varied from far below Emax to around Emax even in the tumor for which use of the drugs had been accepted. These results suggest that rat AUCss,u at the MTD, but not the efficacy in xenograft mice, may be a useful parameter to estimate the human clinical dose of kinase inhibitors, which seems to be currently determined by toxicity rather than efficacy.
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Notes
  • Ryota ISE, Satoshi KONDO, Hiroto KATO, Noritaka IMAI, Hideo AKIYAMA, K ...
    Type: Note
    2011 Volume 26 Issue 6 Pages 621-626
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: August 16, 2011
    JOURNALS FREE ACCESS
      Preclinical data of fetal, infant, and juvenile animals are important for the prediction of drug toxicity in fetuses and children. However, expression of drug-metabolizing enzymes, including cytochromes P450 (CYPs), have not been fully investigated in fetal, infant, or juvenile liver of the cynomolgus macaque, an animal species important for preclinical studies. In this study, hepatic expression of 20 cynomolgus macaque CYPs (mfCYPs) in the CYP1–4 subfamilies that are relevant to drug metabolism was measured in fetuses, infants, and juveniles using DNA microarrays. Expression of most mfCYPs, including those moderately or abundantly expressed in postnatal livers such as mfCYP2A23, mfCYP2A24, mfCYP2B6, mfCYP2C9, mfCYP2C19, mfCYP2C76, mfCYP2D17, mfCYP2E1 mfCYP3A4, and mfCYP3A5, was much less abundant in fetal livers, but increased substantially after birth. In contrast, expression of mfCYP2C8 in fetal livers was not substantially different from postnatal livers. Since human CYP3A7 is expressed more abundantly in fetal livers than in adult livers, mfCYP3A7, an ortholog of human CYP3A7, was analyzed by quantitative polymerase chain reaction. Expression of mfCYP3A7 in fetal livers was much lower than that in postnatal livers, and greatly increased after birth, unlike the expression of human CYP3A7. These results indicate that expression of most mfCYPs examined was low in fetal livers, but increased greatly in postnatal livers, with a few exceptions such as mfCYP2C8.
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  • Yasuhiro UNO, Shotaro UEHARA, Hiroshi YAMAZAKI
    Type: Note
    2011 Volume 26 Issue 6 Pages 627-631
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: August 16, 2011
    JOURNALS FREE ACCESS
      The cytochrome P450 (CYP) 1 family consists of the CYP1A, CYP1B, CYP1C, and CYP1D subfamilies. In humans, CYP1A1, CYP1A2, and CYP1B1 are expressed and encode functional enzymes, whereas CYP1D1P (formerly known as CYP1A8P) is present as a pseudogene as a result of five nonsense mutations in exon 2 and exon 7 of the putative coding region. We previously identified CYP1D1 in macaques and found that it was expressed and functional in liver. Moreover, the nonsense mutations in exon 2 and exon 7 were not found in the 20 cynomolgus macaques and 10 rhesus macaques analyzed in that previous study. These results raised the possibility that CYP1D1 is a functional gene in macaques; however, the possibility that nonsense mutations are present in other exons cannot be excluded. In this study, we sought to identify genetic variants of CYP1D1 in 63 cynomolgus macaques and 30 rhesus macaques; we did not find nonsense mutations in any coding exon of the animals analyzed. Moreover, 15 of the 63 cynomolgus macaques were analyzed by quantitative polymerase chain reaction, confirming hepatic expression of CYP1D1 in all 15 animals. These results suggest that CYP1D1 is most likely functional in cynomolgus macaques and rhesus macaques.
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  • Takeo NAKANISHI, Yoko FUKUYAMA, Masaharu FUJITA, Yoshiyuki SHIRASAKA, ...
    Type: Note
    2011 Volume 26 Issue 6 Pages 632-636
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: October 12, 2011
    JOURNALS FREE ACCESS
      To study transport of the carnitine precursor gamma-butyrobetaine (GBB) by rat liver-specific GABA transporter 2 (rGat2), we measured the uptake of deuterated GBB (d3-GBB) by Xenopus oocytes expressing rGat2. rGat2-mediated d3-GBB uptake was Na+ and Cl dependent, and was saturable with a Km value of 70.6 ± 34.8 µM. In carnitine transporter Octn2-deficient Jvs mice, mRNA expression of mGat3, the mouse ortholog of GAT2, decreased in liver, but increased in brain, while mRNA expression of mBbox1, which hydroxylates GBB to carnitine, increased in kidney and brain. These results indicate that GAT2 plays a role in transmembrane transport of GBB.
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SNP Communications
  • Jonathan D. URBAN, Robert A. BUDINSKY, J. Craig ROWLANDS
    Type: SNP Communication
    2011 Volume 26 Issue 6 Pages 637-645
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: August 09, 2011
    JOURNALS FREE ACCESS
      Species' variation(s) in gene homologues can result in differences among species in their quantitative and qualitative susceptibility and responsiveness to environmental contaminants. In the case of dioxin-like compounds (DLCs), it has been hypothesized that single nucleotide polymorphisms (SNPs) in genes associated with aryl hydrocarbon receptor (AHR)-regulated pathways may result in greater susceptibility to DLC toxicity. A key step in the activation of AHR involves heterodimerization with the AHR nuclear translocator (ARNT) protein before binding to its DNA response element. The objective of this study was to identify SNPs in the human ARNT gene that could potentially affect the sensitivity of AHR-dependent gene transcription. Results from DNA sequencing of 101 human samples demonstrated the presence of five unique SNPs at the ARNT locus, including three non-synonymous SNPs, of which two were novel: V304M and T462A. The genetic frequencies of the non-synonymous SNPs were very low (≤0.02), and the novel SNPs occurred in the Per-ARNT-Sim (PAS) functional domain. In silico analysis indicated that V304M was the only SNP identified in the current population with the potential to significantly alter ARNT protein function. Our findings indicated a very limited occurrence of SNPs with predicted functional consequence in key domains of human ARNT.
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  • Keiko MAEKAWA, Tetsuya HAMAGUCHI, Yoshiro SAITO, Naoko TATEWAKI, Kouic ...
    Type: SNP Communication
    2011 Volume 26 Issue 6 Pages 646-658
    Published: 2011
    Released: December 28, 2011
    [Advance publication] Released: August 16, 2011
    JOURNALS FREE ACCESS
      Glutathione S-transferases (GSTs) play a vital role in the phase II biotransformation of many chemicals, including anticancer drugs. In this study, to elucidate the haplotype structures of the two closely related alpha-class genes GSTA1 and GSTA2, we screened for genetic variation in 214 Japanese colorectal cancer patients who received oxaliplatin-based chemotherapy. By direct resequencing of the 5′-flanking region, all the exons, and their flanking introns for 107 patients, 29 and 27 variants were identified in GSTA1 and GSTA2, respectively. The known functional single nucleotide polymorphisms (SNPs) −567T>G, −69C>T, and −52G>A in GSTA1*B were found at allele frequencies of 0.140. Of the four major GSTA2 allelic variants reported previously (GSTA2*A, *B, *C, and *E), only GSTA2*B (frequency = 0.154), *C (0.706), and *E (0.140) were detected. Following linkage disequilibrium analysis, haplotypes of both genes were separately estimated. Then, rapid genotyping methods for 7 and 6 SNPs tagging common haplotypes of GSTA1 and GSTA2, respectively, were developed using the single-base extension assay, and an additional 107 patients were genotyped. Finally, haplotype combinations of both genes were classified into 3 major types: GSTA1*A-GSTA2*C, GSTA1*A-GSTA2*B, and GSTA1*B-GSTA2*E. These findings will be useful in pharmacogenomic studies on xenobiotics including anticancer drugs.
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