Faculty Profiles - ARIYOSHI Noritaka

Papers - ARIYOSHI Noritaka

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Adverse events of nivolumab plus ipilimumab versus nivolumab plus cabozantinib: a real-world pharmacovigilance study. Reviewed International journal

Yurie Oka, Jun Matsumoto, Tatsuaki Takeda, Naohiro Iwata, Takahiro Niimura, Aya Fukuma Ozaki, Kensuke Bekku, Hirofumi Hamano, Motoo Araki, Keisuke Ishizawa, Yoshito Zamami, Noritaka Ariyoshi

International journal of clinical pharmacy 2024.4

Authorship：Last author Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: No head-to-head clinical trials have compared the differences in adverse events (AEs) between nivolumab plus ipilimumab (NIVO-IPI) and nivolumab plus cabozantinib (NIVO-CABO) in the treatment of metastatic renal cell carcinoma (mRCC). AIM: We analysed the two largest real-world databases, the U.S. Food and Drug Administration Adverse Event Reporting System (FAERS) and the World Health Organization's VigiBase, to elucidate the differences in AEs between NIVO-IPI and NIVO-CABO. METHOD: In total, 40,376 and 38,022 records were extracted from FAERS and VigiBase, and 193 AEs were analysed. The reporting odds ratios (ROR) with 95% confidence interval were calculated using a disproportionality analysis (NIVO-CABO/NIVO-IPI). RESULTS: The reported numbers of immune-related AEs, including myocarditis, colitis, and hepatitis, were significantly higher with NIVO-IPI (ROR = 0.18 for FAERS and 0.13 for VigiBase). Contrarily, the reported numbers of other AEs, including gastrointestinal disorders (ROR = 2.68 and 2.92) and skin and subcutaneous tissue disorders (ROR = 2.94 and 3.55), considered to be potentiated by the combination of NIVO and CABO, were higher with NIVO-CABO. CONCLUSION: Our findings contribute to the selection and clinical management of NIVO-IPI and NIVO-CABO, which minimizes the risk of AEs for individual patients with mRCC by considering distinctive differences in the AE profiles.

DOI： 10.1007/s11096-024-01713-1

A comparison between the adverse event profiles of patients receiving palbociclib and abemaciclib: analysis of two real-world databases. Reviewed International journal

Tatsuaki Takeda, Shiho Sugimoto, Jun Matsumoto, Naohiro Iwata, Akihiko Nakamoto, Aya Fukuma Ozaki, Hirofumi Hamano, Noritaka Ariyoshi, Yoshito Zamami

International journal of clinical pharmacy 2024.1

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Palbociclib and abemaciclib are cyclin-dependent kinase (CDK) 4/6 inhibitors currently used to treat breast cancer. Although their therapeutic efficacies are considered comparable, differences in adverse event (AE) profiles of the two drugs remain unclear. AIM: We analysed two real-world databases, the World Health Organization's VigiBase and the Food and Drug Administration Adverse Event Reporting System (FAERS), to identify differences in AE profiles of palbociclib and abemaciclib. METHOD: Data of patients with breast cancer receiving palbociclib or abemaciclib recorded until December 2022 were extracted from the VigiBase and FAERS databases. In total, 200 types of AEs were analysed. The reporting odds ratios were calculated using a disproportionality analysis. RESULTS: Cytopenia was frequently reported in patients receiving palbociclib, whereas interstitial lung disease and diarrhoea were frequently reported in those receiving abemaciclib. Moreover, psychiatric and nervous system disorders were more common in the palbociclib group, whereas renal and urinary disorders were more common in the abemaciclib group. CONCLUSION: This study is the first to show comprehensively the disparities in the AE profiles of palbociclib and abemaciclib. The findings highlight the importance of considering these differences when selecting a suitable CDK4/6 inhibitor to ensure safe and favourable outcomes for patients with breast cancer.

DOI： 10.1007/s11096-023-01687-6

Significance of UGT1A6, UGT1A9, and UGT2B7 genetic variants and their mRNA expression in the clinical outcome of renal cell carcinoma. Reviewed International journal

Jun Matsumoto, Anzu Nishimoto, Shogo Watari, Hideo Ueki, Shoya Shiromizu, Naohiro Iwata, Tatsuaki Takeda, Soichiro Ushio, Makoto Kajizono, Masachika Fujiyoshi, Toshihiro Koyama, Motoo Araki, Koichiro Wada, Yoshito Zamami, Yasutomo Nasu, Noritaka Ariyoshi

Molecular and cellular biochemistry 478 ( 8 ) 1779 - 1790 2023.8

Authorship：Last author Language：English Publishing type：Research paper (scientific journal)

UDP-glucuronosyltransferase (UGT) metabolizes a number of endogenous and exogenous substrates. Renal cells express high amounts of UGT; however, the significance of UGT in patients with renal cell carcinoma (RCC) remains unknown. In this study, we profile the mRNA expression of UGT subtypes (UGT1A6, UGT1A9, and UGT2B7) and their genetic variants in the kidney tissue of 125 Japanese patients with RCC (Okayama University Hospital, Japan). In addition, we elucidate the association between the UGT variants and UGT mRNA expression levels and clinical outcomes in these patients. The three representative genetic variants, namely, UGT1A6 541A > G, UGT1A9 i399C > T, and UGT2B7-161C > T, were genotyped, and their mRNA expression levels in each tissue were determined. We found that the mRNA expression of the three UGTs (UGT1A6, UGT1A9, and UGT2B7) are significantly downregulated in RCC tissues. Moreover, in patients with RCC, the UGT2B7-161C > T variant and high UGT2B7 mRNA expression are significantly correlated with preferable cancer-specific survival (CSS) and overall survival (OS), respectively. As such, the UGT2B7-161C > T variant and UGT2B7 mRNA expression level were identified as significant independent prognostic factors of CSS and CSS/OS, respectively. Taken together, these findings indicate that UGT2B7 has a role in RCC progression and may, therefore, represent a potential prognostic biomarker for patients with RCC.

DOI： 10.1007/s11010-022-04637-4

Derivation of a Novel Scoring System Predicting High Platelet Reactivity on Prasugrel in Patients with Coronary Artery Disease Reviewed

Yuichi Saito, Takeshi Nishi, Shinichi Wakabayashi, Yuji Ohno, Hideki Kitahara, Noritaka Ariyoshi, Yoshio Kobayashi

Journal of Atherosclerosis and Thrombosis 29 ( 11 ) 1625 - 1633 2022.11

Adverse Events of Axitinib plus Pembrolizumab Versus Lenvatinib plus Pembrolizumab: A Pharmacovigilance Study in Food and Drug Administration Adverse Event Reporting System. Reviewed International journal

Jun Matsumoto, Naohiro Iwata, Shogo Watari, Soichiro Ushio, Shoya Shiromizu, Tatsuaki Takeda, Hirofumi Hamano, Makoto Kajizono, Motoo Araki, Yasutomo Nasu, Noritaka Ariyoshi, Yoshito Zamami

European urology focus 9 ( 1 ) 141 - 144 2022.7

Language：English Publishing type：Research paper (scientific journal)

No head-to-head postmarket surveillance study has compared the differences in adverse events (AEs) between two combination therapies, axitinib (AXI) + pembrolizumab (PEMBRO) and lenvatinib (LEN) + PEMBRO, against metastatic renal cell carcinoma. This study aims to highlight the comprehensive differences in AEs between these two therapies based on the real-world big data from the Food and Drug Administration Adverse Event Reporting System (FAERS) database. In total, 28 937 records were extracted from the FAERS database, and 139 AEs grouped into the System Organ Class according to the Medical Dictionary for Regulatory Activities were analysed. Logistic regression analyses were performed, and the reporting odds ratio with a 95% confidence interval was determined. We found that the incidences of cardiac and hepatobiliary disorders for AXI + PEMBRO, and blood and lymphatic system, metabolism and nutrition, and vascular disorders for LEN + PEMBRO, all of which were associated with serious AEs, were higher than those for LEN + PEMBRO and AXI + PEMBRO, respectively. The differences in the AEs between AXI + PEMBRO and LEN + PEMBRO were not derived merely from those between AXI and LEN monotherapies. Furthermore, remarkable AE potentiation was observed for AXI + PEMBRO. As FAERS is a spontaneous reporting system comprising partially limited information, analysing more detailed relationships between AEs and patient or treatment characteristics was challenging in this study. The present study is the first to show the overall real-world postmarketing differences in AEs between AXI + PEMBRO and LEN + PEMBRO. Our novel findings will substantially improve clinical practice; we recommend comparing patients' conditions associated with the above AEs when selecting between these two therapies. PATIENT SUMMARY: Herein, we highlight the differences in adverse events (AEs) between axitinib + pembrolizumab and lenvatinib + pembrolizumab therapies using data from the real-world Food and Drug Administration Adverse Event Reporting System database aimed at patients with metastatic renal cell carcinoma. We identified AEs that needed attention in each combination. We recommend the differences in AEs to be considered when selecting these two therapies.

DOI： 10.1016/j.euf.2022.07.003

Differential Impact of Clinical and Genetic Factors on High Platelet Reactivity in Patients with Coronary Artery Disease Treated with Clopidogrel and Prasugrel. Reviewed

Yuichi Saito, Takeshi Nishi, Shinichi Wakabayashi, Yuji Ohno, Hideki Kitahara, Noritaka Ariyoshi, Yoshio Kobayashi

Journal of atherosclerosis and thrombosis 29 ( 7 ) 1031 - 1039 2022.7

Language：English Publishing type：Research paper (scientific journal)

AIM: High platelet reactivity (HPR) is associated with increased risks of thrombotic events in patients with coronary artery disease. The recently developed ABCD-GENE score identified five clinical and genetic factors (age, body mass index, chronic kidney disease, diabetes, and the CYP2C19 loss-of-function allele) for HPR, although the significance of various stages of each factor is unclear. METHODS: Four prospective studies were pooled, in which platelet reactivity was measured using the VerifyNow assay with clopidogrel and prasugrel; genotyping of CYP2C19 was also performed. Each component of the ABCD-GENE score was divided into three subcategories. VerifyNow P2Y12 reactivity units ＞208 were defined as HPR. RESULTS: A total of 184 patients were included, of which 111 (60%) and 51 (28%) had HPR with clopidogrel and prasugrel. Chronic kidney disease had an impact on HPR on both clopidogrel and prasugrel, whereas the impact of diabetes was more evident in patients treated with prasugrel. Although the number of CYP2C19 loss-of-function alleles was clearly associated with a likelihood of HPR with clopidogrel, P2Y12 reactivity units with prasugrel treatment were also significantly and progressively higher in patients with more CYP2C19 loss-of-function alleles. CONCLUSIONS: Clinical and genetic factors had a differential effect on a P2Y12 inhibitor reactivity with clopidogrel and prasugrel in patients with coronary artery disease. The severity of the factors also had a different impact on HPR.

DOI： 10.5551/jat.63035

Blood concentrations of tacrolimus upon conversion from rabeprazole to vonoprazan in renal transplant recipients: Correlation with cytochrome P450 gene polymorphisms. Reviewed International journal

Shogo Watari, Motoo Araki, Jun Matsumoto, Kasumi Yoshinaga, Takanori Sekito, Yuki Maruyama, Yosuke Mitsui, Takuya Sadahira, Risa Kubota, Shingo Nishimura, Koichiro Wada, Yasuyuki Kobayashi, Hidemi Takeuchi, Katsuyuki Tanabe, Masashi Kitagawa, Hiroshi Morinaga, Shinji Kitamura, Hitoshi Sugiyama, Noritaka Ariyoshi, Jun Wada, Masami Watanabe, Toyohiko Watanabe, Yasutomo Nasu

Drug metabolism and pharmacokinetics 40 100407 - 100407 2021.10

Language：English Publishing type：Research paper (scientific journal)

We evaluated the impact of vonoprazan on blood concentrations of tacrolimus via a retrospective analysis of 52 renal transplant recipients who took tacrolimus and converted from rabeprazole to vonoprazan between August 2018 and September 2019. We compared tacrolimus trough levels upon conversion among groups that were classified based on cytochrome P450 (CYP) gene polymorphisms. CYP3A5 groups were heterozygous or homozygous for CYP3A5∗1 and CYP3A5∗3 alleles. CYP2C19 genotypes were classified as extensive (∗1/∗1), intermediate (∗1/∗2 and ∗1/∗3) or poor metabolizers (∗2/∗2, ∗2/∗3 and ∗3/∗3). Tacrolimus trough levels increased only 0.3 ng/mL upon conversion in the CYP3A5∗3/∗3 group: 5.8 [3.4-7.2] vs 6.1 [3.8-7.9]; p = 0.06. No statistically significance changes in tacrolimus levels also occurred in the CYP3A5∗1/∗1 or CYP3A5∗1/∗3 groups. Subgroup analyses of CYP3A5∗3/∗3 demonstrated low changes for all three CYP2C19 subgroups: 5.2 [4.3-6.5] vs 6.2 [4.3-7.9]; p = 0.07, 6.1 [3.4-7.2] vs 6.7 [4.6-7.9]; p = 0.12 and 5.4 [3.6-6.5] vs 4.7 [3.8-6.3]; p = 1.00, respectively. Conversion to vonoprazan thus resulted in little increase of tacrolimus trough levels, even in the group predicted to be most susceptible (CYP3A5∗3/∗3 and 2C19∗1/∗1), thus supporting the safety of concomitant use of vonoprazan with tacrolimus.

DOI： 10.1016/j.dmpk.2021.100407

Relevance of CYP3A5 Expression on the Clinical Outcome of Patients With Renal Cell Carcinoma. Reviewed International journal

Jun Matsumoto, Yumi Kotera, Shogo Watari, Koichi Takeuchi, Hideo Ueki, Toshihiro Koyama, Koichiro Wada, Masachika Fujiyoshi, Yasutomo Nasu, Noritaka Ariyoshi

Anticancer research 41 ( 5 ) 2511 - 2521 2021.5

Authorship：Last author Language：English Publishing type：Research paper (scientific journal)

BACKGROUND/AIM: This study aimed to elucidate the detailed characteristics of CYP3A5 expression and the association between CYP3A5 expression and clinical outcomes in patients with renal cell carcinoma (RCC). PATIENTS AND METHODS: This study retrospectively enrolled 124 Japanese patients with RCC treated at the Okayama University Hospital. The commonest CYP3A5 gene polymorphism, CYP3A5*3, and expression levels of CYP3A5 mRNA and protein in each tissue were examined. RESULTS: Expression of CYP3A5 mRNA and protein in RCC tissues was significantly down-regulated compared to that in adjacent normal tissues. High level of CYP3A5 mRNA expression significantly extended cancer-specific survival (p=0.004) and overall survival (p=0.002). The CYP3A5 mRNA expression level was identified as a significant independent prognostic factor for both cancer-specific survival and overall survival. CONCLUSION: CYP3A5 could serve as a potential marker for prognostication and treatment planning for patients with RCC.

DOI： 10.21873/anticanres.15029

Validation of the ABCD-GENE score to identify high platelet reactivity in east Asian patients undergoing percutaneous coronary intervention. Reviewed International journal

Yuichi Saito, Takeshi Nishi, Shinichi Wakabayashi, Yuji Ohno, Hideki Kitahara, Noritaka Ariyoshi, Yoshio Kobayashi

International journal of cardiology 327 15 - 18 2021.3

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: High platelet reactivity (HPR) is associated with subsequent thrombotic events in patients undergoing percutaneous coronary intervention (PCI). Recently, the ABCD-GENE score was developed to identify patients at risk for HPR, incorporating both clinical and genetic factors. However, this score was derived and validated in mostly Caucasian subjects and it has not been validated in an East Asian population. METHOD: Individual patient data from 4 prospective studies were pooled, in which platelet reactivity was measured using the VerifyNow assay on clopidogrel and genotyping of CYP2C19 was performed after PCI. Study populations included patients with general stable coronary artery disease, hemodialysis, age ≥75 and/or body weight <50 kg, and acute coronary syndrome. VerifyNow P2Y12 reactivity units >208 was defined as HPR. RESULTS: Of 184 patients, 111 (60%) had HPR on clopidogrel. In the receiver operating characteristics curve analyses, the ABCD-GENE score significantly predicted HPR on clopidogrel (AUC 0.78, best cut-off value 9, p < 0.001). Across the 4 studies and their combinations, the diagnostic ability and cut-off values of ABCD-GENE score for HPR on clopidogrel were consistent. CONCLUSIONS: The ABCD-GENE score had significant and moderate diagnostic ability for HPR on clopidogrel in Japanese patients undergoing PCI. The predictivity was consistent across a broad spectrum of patient populations, suggesting the applicability of this novel scoring system in clinical practice worldwide.

DOI： 10.1016/j.ijcard.2020.11.022

【コロナ禍における医療人育成】薬学領域コロナ禍における薬学実務実習での工夫

猪田宏美, 藤吉正哉, 西原茂樹, 松本准, 有吉範高, 千堂年昭

医学教育 51 ( 5 ) 541 - 543 2020.10

Effect of CYP3A5*3 genetic variant on the metabolism of direct-acting antivirals in vitro: a different effect on asunaprevir versus daclatasvir and beclabuvir. Reviewed International journal

Jun Matsumoto, Su Nwe San, Masachika Fujiyoshi, Ayano Kawauchi, Natsumi Chiba, Ran Tagai, Ryoko Sanbe, Shiho Yanaka, Hiroaki Sakaue, Yoshinori Kato, Hiroyoshi Nakamura, Harumi Yamada, Noritaka Ariyoshi

Journal of human genetics 65 ( 2 ) 143 - 153 2020.1

Authorship：Last author Language：English Publishing type：Research paper (scientific journal)

Direct-acting antivirals, asunaprevir (ASV), daclatasvir (DCV), and beclabuvir (BCV) are known to be mainly metabolized by CYP3A enzymes; however, the differences in the detailed metabolic activities of CYP3A4 and CYP3A5 on these drugs are not well clarified. The aim of the present study was to elucidate the relative contributions of CYP3A4 and CYP3A5 to the metabolism of ASV, DCV, and BCV, as well as the effect of CYP3A5*3 genetic variant in vitro. The amount of each drug and their major metabolites were determined using LC-MS/MS. Recombinant CYP3As and CYP3A5*3-genotyped human liver microsomes (CYP3A5 expressers or non-expressers) were used for the determination of their metabolic activities. The contribution of CYP3A5 to ASV metabolism was considerable compared to that of CYP3A4. Consistently, ASV metabolic activity in CYP3A5 expressers was higher than those in CYP3A5 non-expresser. Moreover, CYP3A5 expression level was significantly correlated with ASV metabolism. In contrast, these observations were not found in DCV and BCV metabolism. To our knowledge, this is the first study to directly demonstrate the effect of CYP3A5*3 genetic variants on the metabolism of ASV. The findings of the present study may provide basic information on ASV, DCV, and BCV metabolisms.

DOI： 10.1038/s10038-019-0685-2

A randomized controlled, open-label early phase II trial comparing incidence of FOLFIRI.3-induced diarrhoea between Hangeshashinto and oral alkalization in Japanese patients with colorectal cancer. Reviewed International journal

Kaori Yamazaki, Noritaka Ariyoshi, Hideaki Miyauchi, Gaku Ohira, Noriko Kaneya, Kohei Yamamoto, Kenichi Arai, Shingo Yamazaki, Hisahiro Matsubara, Takaaki Suzuki, Itsuko Ishii

Journal of clinical pharmacy and therapeutics 44 ( 6 ) 946 - 951 2019.12

Language：English Publishing type：Research paper (scientific journal)

WHAT IS KNOWN AND OBJECTIVE: We conducted a pilot clinical trial to investigate whether Hangeshashinto (TJ-14) could be substituted for oral alkalization in patients scheduled to undergo chemotherapy by FOLFIRI.3 regimen for colorectal cancer (CRC). METHODS: Patients with CRC were randomized 1:1 to a TJ-14 (7.5 g/day) group or an oral alkalization (sodium bicarbonate, 1.8 g/day; ursodeoxycholic acid, 300 mg/day) group. The primary endpoint was incident of late-onset diarrhoea. A total of 30 patients were randomized to either the TJ-14 group or the alkalization group. RESULTS AND DISCUSSION: There was no statistical difference in age, concomitantly used drugs or UGT1A1 genotypes between the groups. In the alkalization group (n = 15), the frequency of grade 0/1/2 and grade 3 diarrhoea was 73% and 27%, respectively. In the TJ-14 group (n = 14), the frequency of grade 0/1/2 and grade 3 diarrhoea was 79% and 21%, respectively. Grade 4 diarrhoea was not observed in either group. There was no statistically significant difference in other adverse events or in response to FOLFIRI.3 between the groups. WHAT IS NEW AND CONCLUSION: This pilot trial suggests that TJ-14 is a promising alternative treatment option to reduce FOLFIRI.3-induced late-onset diarrhoea, although additional clinical study with a larger number of patients is necessary to confirm these results.

DOI： 10.1111/jcpt.13020

Minor contribution of CYP3A5 to the metabolism of hepatitis C protease inhibitor paritaprevir in vitro. Reviewed International journal

Su Nwe San, Jun Matsumoto, Yumi Saito, Masako Koike, Hiroaki Sakaue, Yoshinori Kato, Masachika Fujiyoshi, Noritaka Ariyoshi, Harumi Yamada

Xenobiotica; the fate of foreign compounds in biological systems 49 ( 8 ) 935 - 944 2019.8

Language：English Publishing type：Research paper (scientific journal)

Paritaprevir (PTV) is a non-structural protein 3/4A protease inhibitor developed for the treatment of hepatitis C disease as a fixed dose combination of ombitasvir (OBV) and ritonavir (RTV) with or without dasabuvir. The aim of this study was to evaluate the effects of cytochrome P450 (CYP) 3A5 on in vitro PTV metabolism using human recombinant CYP3A4, CYP3A5 (rCYP3A4, rCYP3A5) and human liver microsomes (HLMs) genotyped as either CYP3A5*1/*1, CYP3A5*1/*3 or CYP3A5*3/*3. The intrinsic clearance (CLint, Vmax/Km) for the production of a metabolite from PTV in rCYP3A4 was 1.5 times higher than that in rCYP3A5. The PTV metabolism in CYP3A5*1/*1 and CYP3A5*1/*3 HLMs expressing CYP3A5 was comparable to that in CYP3A5*3/*3 HLMs, which lack CYP3A5. CYP3A4 expression level was significantly correlated with PTV disappearance rate and metabolite formation. In contrast, there was no such correlation found for CYP3A5 expression level. This study represents that the major CYP isoform involved in PTV metabolism is CYP3A4, with CYP3A5 having a minor role in PTV metabolism. The findings of the present study may provide foundational information on PTV metabolism, and may further support dosing practices in HCV-infected patients prescribed PTV-based therapy.

DOI： 10.1080/00498254.2018.1524947

A proposed simple screening method to determine relative contributions of CYP3A4 and CYP3A5 to drug metabolism in vitro. Reviewed

Matsumoto J, Nakamura H, San NS, Sato H, Takezawa M, Kishi R, Kito Y, Sugano J, Izuki M, Yanagisawa N, Ikeda N, Saito Y, Kato Y, Yamada H, Fujiyoshi M, Ariyoshi N

Pers Med Univers. 8 41 - 44 2019.7

High residual platelet reactivity after switching from clopidogrel to low-dose prasugrel in Japanese patients with end-stage renal disease on hemodialysis. Reviewed International journal

Yuji Ohno, Hideki Kitahara, Kenichi Fujii, Yukinori Kohno, Noritaka Ariyoshi, Takeshi Nishi, Yoshihide Fujimoto, Yoshio Kobayashi

Journal of cardiology 73 ( 1 ) 51 - 57 2019.1

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: High on-treatment platelet reactivity (HPR) under clopidogrel treatment is frequently observed in hemodialysis (HD) patients. In such patients, 10mg of prasugrel has reportedly inhibited platelet reactivity more adequately compared with 75mg of clopidogrel. However, the efficacy of 3.75mg prasugrel in Japanese HD patients is largely unknown. METHODS: A total of 41 Japanese coronary artery disease patients under HD who received aspirin and clopidogrel were enrolled. Clopidogrel was switched to 3.75mg prasugrel. At day 14, prasugrel was switched to clopidogrel. Platelet reactivity was measured using VerifyNow assay (Accumetrics, San Diego, CA, USA) at baseline, day 14, and day 28. VerifyNow P2Y12 reaction units (PRU) >208 was defined as HPR. RESULTS: The PRU level on prasugrel therapy was significantly lower than that on clopidogrel therapy before switching (219.1±62.3 PRU vs. 238.2±68.0 PRU, p=0.02). Although the prevalence of HPR was numerically lower on prasugrel therapy compared with clopidogrel therapy before and after switching, the differences did not reach a statistical significance (57.6% vs. 75.7% vs. 74.2%, p=0.13). Even under prasugrel treatment, more than half of patients showed HPR. CONCLUSIONS: Although low-dose prasugrel had somewhat better antiplatelet effect than clopidogrel, it could not significantly improve the prevalence of HPR in Japanese HD patients. Higher doses of prasugrel might be needed to achieve adequate platelet inhibition in this high thrombotic risk population.

DOI： 10.1016/j.jjcc.2018.07.001

Efficacy of 2.5-mg Prasugrel in Elderly or Low-Body-Weight Patients. Reviewed

Shinichi Wakabayashi, Noritaka Ariyoshi, Hideki Kitahara, Kenichi Fujii, Yoshihide Fujimoto, Yoshio Kobayashi

Circulation journal : official journal of the Japanese Circulation Society 82 ( 9 ) 2326 - 2331 2018.8

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Due to concern about bleeding complications, a maintenance dose of prasugrel 2.5 mg may be used in elderly or low-body-weight patients in Japan. There is little information, however, on the efficacy and safety of a 2.5-mg maintenance dose of prasugrel. Methods and Results: In this single-center, prospective, open-label, cross-over study, a total of 44 elderly (≥75 years old) or low body-weight (<50 kg) Japanese patients >1 month after percutaneous coronary intervention who were treated with aspirin 81-100 mg and clopidogrel 75 mg were randomized to either prasugrel 2.5 mg or 3.75 mg instead of clopidogrel for 14 days, with a cross-over directly to the alternate treatment for another 14 days. Platelet inhibition was assessed with the VerifyNow assay (Accumetrics, San Diego, CA, USA) at 3 time points: baseline; day 14; and day 28. P2Y12 reaction units (PRU) ≤95 was defined as low on-treatment platelet reactivity (LPR), and PRU ≥262 as high on-treatment platelet reactivity (HPR). The prevalence of LPR was 2.2% in patients treated with clopidogrel, 2.2% in those with prasugrel 2.5 mg, and 22.7% in those with prasugrel 3.75 mg (P<0.001). Clopidogrel resulted in the higher prevalence of HPR compared with 2.5-mg and 3.75-mg prasugrel (40.9% vs. 18.2% vs. 6.8%, P<0.001). CONCLUSIONS: Prasugrel 2.5 mg may be more appropriate in elderly or lower-body-weight Japanese patients.

DOI： 10.1253/circj.CJ-18-0337

Platelet inhibition after loading dose of prasugrel in patients with ST-elevation and non-ST-elevation acute coronary syndrome. Reviewed

Shinichi Wakabayashi, Hideki Kitahara, Takeshi Nishi, Kazumasa Sugimoto, Takashi Nakayama, Yoshihide Fujimoto, Noritaka Ariyoshi, Yoshio Kobayashi

Cardiovascular intervention and therapeutics 33 ( 3 ) 239 - 246 2018.7

Language：English Publishing type：Research paper (scientific journal)

The PRASFIT-ACS study showed the antiplatelet effect 2-4 h after prasugrel loading. However, there is little information about the antiplatelet effect <2 h after prasugrel loading dose, especially in patients with acute coronary syndrome (ACS). There had not been any comparison between ST-segment elevation myocardial infarction (STEMI) and non-ST-segment elevation ACS (NSTE-ACS). Fifty patients with ACS (15 with STEMI and 35 with NSTE-ACS) were enrolled. They received a 20-mg prasugrel loading dose followed by a maintenance dose of 3.75 mg. Platelet reactivity [P2Y12 reaction units (PRU)] was evaluated by the VerifyNow assay at baseline, 30 min, 1, 2, 4, and 6 h, 1 week under prasugrel, and at 1 month after switching to clopidogrel. The primary end point was the change of PRU compared to the baseline. Furthermore, PRU after prasugrel loading between STEMI and NSTE-ACS was compared. A significant reduction in PRU from baseline was observed at ≥2 h after administration of prasugrel. In STEMI patients, a significant reduction in PRU was observed at 4 h after prasugrel loading. STEMI patients had higher PRU compared to NSTE-ACS patients at 2, 4, and 6 h after prasugrel loading. Utilizing >208 PRU as a cutoff value, STEMI patients had a higher prevalence of high on-treatment platelet reactivity at 2-6 h and 1 week after loading. Rapid antiplatelet effect is not achieved by low-dose prasugrel loading in STEMI patients. Platelet inhibition occurs earlier in NSTE-ACS patients, although it takes ≥2 h after loading in the majority of patients.

DOI： 10.1007/s12928-017-0475-8

HPLC-UVを用いたソホスブビルの定量法の確立(Determination of sofosbuvir via a high-performance liquid chromatography method using ultraviolet detection)

Su Nwe San, 松本准, 小池雅子, 斎藤祐実, 坂上弘明, 加藤芳徳, 有吉範高, 山田治美

国際医療福祉大学学会誌 23 ( 1 ) 130 - 136 2018.3

Language：English Publisher：国際医療福祉大学学会

Sofosbuvir(SOF)は、C型肝炎ウイルスの増殖を阻害するヌクレオチドアナログ薬である。本研究において、我々は、水溶液およびヒト血漿中SOF濃度の簡便、特異的、高感度および精密なUV検出を用いた高速クロマトグラフィー法による測定方法の確立を試みた。0.25mLの水溶液またはヒト血漿からアセトニトリルを含む抽出液で抽出し、C18逆相カラムを用いて分析した。ソラフェニブを内標準物質(IS)とし、移動相は酢酸アンモニウム緩衝液、メタノールおよびアセトニトリルを用いた。流速は1.0mL/min、UV検出波長は265nmとした。SOFとISの検出時間はそれぞれ7.8minおよび9.1min、1検体の測定時間は15.0minであった。検量線は、0.1-10.0μg/mLの範囲で直線性を得た。それぞれの検出限界は0.1μg/mL、ヒト血漿試料からの回収率は99%以上であった。水溶液およびヒト血漿中の日内、日間変動係数はそれぞれ1.41-10.06%および0.21-6.53%であった。日内、日間精度はそれぞれ92.71-98.41%および89.31-101.21%であった。本方法は、非常に簡便でかつ試料から良好な回収が可能でありSOFの薬物動態研究を行うために用いることができる。(著者抄録)

Other Link： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2018&ichushi_jid=J05958&link_issn=&doc_id=20180426420014&doc_link_id=http%3A%2F%2Fid.nii.ac.jp%2F1065%2F00000857%2F&url=http%3A%2F%2Fid.nii.ac.jp%2F1065%2F00000857%2F&type=%8D%91%8D%DB%88%E3%97%C3%95%9F%8E%83%91%E5%8Aw%81F%8D%91%8D%DB%88%E3%97%C3%95%9F%8E%83%91%E5%8Aw%83%8A%83%7C%83W%83g%83%8A&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F80077_3.gif

An extremely high bioavailability of orally administered vancomycin in a patient with severe colitis and renal insufficiency Reviewed

Shingo Yamazaki, Takaaki Suzuki, Tatsuya Suzuki, Hirokazu Takatsuka, Masayuki Ishikawa, Noriyuki Hattori, Takeshi Fujishiro, Hideaki Miyauchi, Takehiko Oami, Noritaka Ariyoshi, Shigeto Oda, Hisahiro Matsubara, Itsuko Ishii

JOURNAL OF INFECTION AND CHEMOTHERAPY 23 ( 12 ) 848 - 851 2017.12

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

Because there is little absorption of orally administered vancomycin hydrochloride (VCM) through the normal intestinal microvillus membrane, the pharmacokinetics of VCM absorbed from the digestive tract are mostly unknown. Here we report a case of severe colitis and renal insufficiency in which the serum concentration of VCM reached the supratherapeutic range after oral administration. A 54-year-old man receiving outpatient chemotherapy for rectal cancer was admitted to our hospital for severe sepsis and acute renal failure. Multimodal therapy including continuous renal replacement therapy (CRRT) and mechanical ventilation was initiated, and oral VCM administration (0.5 g every 6 h) was begun for suspected severe pseudomembranous colitis with large amounts of watery stool. Despite continued CRRT, the serum VCM concentration increased to 30.6 mg/mL after 4 days. Based on pharmacokinetic analysis, the bioavailability of VCM was estimated to be over 54.5%. Colonoscopy showed that the mucosa was severely damaged throughout the large intestine, resulting in considerable exudation of plasma and blood. This case indicates the need for careful and early monitoring during high-dose oral VCM administration to patients with severe mucosal injury and renal insufficiency. (C) 2017 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.jiac.2017.08.004

Impact of chronic kidney disease on platelet inhibition of clopidogrel and prasugrel in Japanese patients. Reviewed International journal

Takeshi Nishi, Noritaka Ariyoshi, Takashi Nakayama, Yoshihide Fujimoto, Kazumasa Sugimoto, Shinichi Wakabayashi, Hideki Hanaoka, Yoshio Kobayashi

Journal of cardiology 69 ( 5 ) 752 - 755 2017.5

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: The impact of chronic kidney disease (CKD) on the antiplatelet effect of clopidogrel and low-dose (3.75mg) prasugrel in Japanese patients is largely unknown. METHODS: A total of 53 consecutive Japanese patients with stable coronary artery disease who received aspirin and clopidogrel were enrolled, and categorized by estimated glomerular filtration rate (eGFR): CKD group (n=15, eGFR<60ml/min/1.73m2) and non-CKD group (n=38, eGFR≥60ml/min/1.73m2). Clopidogrel was switched to 3.75mg prasugrel. Platelet reactivity measurement using the VerifyNow P2Y12 assay (Accumetrics, San Diego, CA, USA) was performed at baseline (on clopidogrel) and day 14 (on prasugrel). RESULTS: The VerifyNow P2Y12 reaction units (PRU) during clopidogrel therapy was significantly higher in the CKD group than that in the non-CKD group (185.2±51.1 PRU vs. 224.3±57.0 PRU, p=0.02), whereas, the PRU with the prasugrel therapy in the CKD group and non-CKD group were not significantly different (149.9±51.1 PRU vs. 165.3±61.8 PRU, p=0.36). The PRU was significantly lower with the prasugrel therapy compared to that with the clopidogrel therapy both in the CKD group and in the non-CKD group. CONCLUSIONS: Antiplatelet effect of clopidogrel but not prasugrel is attenuated in patients with CKD. Prasugrel achieves a consistently lower platelet reactivity compared with clopidogrel regardless of the presence of mild to moderate CKD.

DOI： 10.1016/j.jjcc.2016.07.017

プロトコルに基づく外来処方の問い合わせの効率化とその効果

内田雅士, 新井さやか, 山崎香織, 竹田真理子, 鈴木貴明, 中村貴子, 有吉範高, 石井伊都子

日本病院薬剤師会雑誌 53 ( 4 ) 417 - 422 2017.4

Language：Japanese Publisher：(一社)日本病院薬剤師会

千葉大学医学部附属病院では院外処方せんに関する問い合わせは薬剤部医薬品情報(DI)室を介して実施している。保険薬局からの問い合わせのうち約90%は医師に照会を行っているが、形式的に医師に確認をとる程度の軽微な内容も多く、医師・患者・保険薬局の負担となっていた。そこで、11項目の問い合わせ内容について問い合わせの受付時に医師に照会を行わず薬剤師の判断で対応するプロトコルを作成し、医師の合意の下、運用を開始した。プロトコル導入後、医師に照会することなく対応した問い合わせは全体の31.1%であり医師に照会した件数は11件/日減少した。医師への疑義照会の所要時間は中央値3.5分(値域1〜237分)であり、保険薬局・患者の待ち時間が短縮される事例があると考えられた。よってプロトコルに基づく運用は有用だと考えられた。(著者抄録)

Other Link： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2017&ichushi_jid=J04451&link_issn=&doc_id=20170414280005&doc_link_id=%2Fdg4hppha%2F2017%2F005304%2F003%2F0417-0422%26dl%3D0&url=https%3A%2F%2Fwww.medicalonline.jp%2Fjamas.php%3FGoodsID%3D%2Fdg4hppha%2F2017%2F005304%2F003%2F0417-0422%26dl%3D0&type=MedicalOnline&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00004_2.gif

院内処方せんに導入した医薬品別検査値表示方式とその有用性 Reviewed

横山威一郎, 橋本杏里, 山口洪樹, 山崎香織, 松島徹, 中村貴子, 鈴木貴明, 有吉範高, 石井伊都子

医療薬学 42 ( 11 ) 738 - 745 2016.11

Language：Japanese Publisher：(一社)日本医療薬学会

本邦で承認されている全ての医療用医薬品の警告・禁忌・原則禁忌(禁忌等)を内容ごとに分類し、処方鑑査での確認可能項目を調査した。医療用医薬品17756品目について添付文書に記載されている禁忌等は51804件で、内容ごとに分類すると14種類となった。禁忌等分類は、疾患・症状の件数が最も多く、半数を占めた。直接臨床検査値と結びつく、肝障害、臨床検査値、腎障害、透析の件数は5291件であり、全体の10.2%であった。確認可能項目は、処方せんに検査値を表示する前は7種類16483件であったのが、検査値表示開始後は11種類21774件となった。処方せんで確認できない項目は3種類30030件あり、全体の58.0%を占めた。採用薬2122品目のうち、禁忌等に該当は287品目505件、「腎」マークの表示に該当は75品目であった。臨床検査値関連の疑義照会によって処方変更に至った事例のうち禁忌に該当する事例は、医薬品別検査値表示開始前は6ヵ月間で0件であったが、表示開始直後は4件、最近の6ヵ月間では20件であった。

Other Link： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2016&ichushi_jid=J03520&link_issn=&doc_id=20161125520003&doc_link_id=%2Fdb5pharm%2F2016%2F004211%2F003%2F0738-0745%26dl%3D0&url=https%3A%2F%2Fwww.medicalonline.jp%2Fjamas.php%3FGoodsID%3D%2Fdb5pharm%2F2016%2F004211%2F003%2F0738-0745%26dl%3D0&type=MedicalOnline&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00004_2.gif

Beta-migrating very low-density lipoprotein conjugates with acrolein in high-cholesterol diet-fed rabbits and localizes to atherosclerotic lesions with macrophages Reviewed

Kanogawa, Yuri, Fujiyoshi, Masachika, Nakazato, Yuki, Watanabe, Kenta, Kurihara, Mizuki, Takezawa, Akari, Uchida, Masashi, Igarashi, Kazuei, Suzuki, Takaaki, Ariyoshi, Noritaka, Ishii, Itsuko

INTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL PATHOLOGY 9 ( 11 ) 11149 - 11158 2016

Language：English Publishing type：Research paper (scientific journal) Publisher：E-CENTURY PUBLISHING CORP

Protein-conjugated acrolein (PC-Acro) is detected in atherosclerotic lesions, and we demonstrated previously that acrolein-conjugated low-density lipoproteins induce macrophage foam cell formation. Although it has been suggested that beta-migrating very low-density lipoprotein (beta VLDL) is taken up by macrophages during atherogenesis, the modification of beta VLDL with acrolein and its localization on lesions are still unclear. The purpose of this study was to clarify the localization of PC-Acro in atherosclerotic lesions and to determine the role of acrolein-conjugated beta VLDL in atherogenesis. Male New Zealand white rabbits were fed 0.5% cholesterol-containing rabbit chow for 8 weeks, and used as an animal model of atherosclerosis. PC-Acro and malondialdehyde (MDA)-conjugated protein levels, which has been used widely as a means to detect oxidized low-density lipoprotein (LDL), in plasma were increased in the 0.5% cholesterol-containing diet-induced animal model of atherosclerosis, whereas their level was unchanged in the control diet fed rabbit. PC-Acro was detected in beta VLDL by western blot analysis, and acrolein-conjugated beta VLDL was effectively taken up by THP-1 macrophages. By immunohistochemical analysis, PC-Acro and macrophages were detected at the internal elastic lamina of the aorta, which was the initial lesion of atherosclerosis. These results suggest that acrolein-conjugated beta VLDL has an important role in the initiation of atherosclerosis via the induction of macrophage foam cell formation in the atherosclerotic lesion.

Organic anion transporting polypeptide 2B1 expression correlates with uptake of estrone-3-sulfate and cell proliferation in estrogen receptor-positive breast cancer cells Reviewed

Jun Matsumoto, Noritaka Ariyoshi, Masahiro Sakakibara, Takeo Nakanishi, Yoshiyuki Okubo, Nobumitsu Shiina, Kaoru Fujisaki, Takeshi Nagashima, Yukio Nakatani, Ikumi Tamai, Harumi Yamada, Hiroshi Takeda, Itsuko Ishii

Drug Metab Pharmacokinet . 30 ( 2 ) 133 - 141 2015.4

Increased platelet inhibition after switching from maintenance clopidogrel to prasugrel in Japanese patients with stable coronary artery disease. Reviewed

Takeshi Nishi, Noritaka Ariyoshi, Takashi Nakayama, Yoshihide Fujimoto, Kazumasa Sugimoto, Masayuki Takahara, Shinichi Wakabayashi, Masaya Koshizaka, Hideki Hanaoka, Yoshio Kobayashi

Circulation journal : official journal of the Japanese Circulation Society 79 ( 11 ) 2439 - 44 2015

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: The pharmacodynamic effects of changing from standard-dose clopidogrel to low-dose (3.75 mg) prasugrel in Japanese patients are largely unknown. METHODS AND RESULTS: A total of 53 consecutive Japanese patients with stable coronary artery disease (CAD) who received aspirin and clopidogrel were enrolled. Clopidogrel was switched to 3.75 mg prasugrel. At day 14, prasugrel was switched to 75 mg clopidogrel. Platelet reactivity was measured using the VerifyNow assay at baseline, day 14, and day 28. VerifyNow P2Y12 reaction units (PRU) >208 was defined as high on-treatment platelet reactivity (HPR). The prevalence of HPR (18.9% vs. 41.5% vs. 44.2%, P<0.001) and the PRU level (154.3±54.2 vs. 196.2±55.5 vs. 194.6±55.8, P<0.001) were significantly lower on prasugrel maintenance therapy compared with the clopidogrel therapy before and after switching. The CYP2C19 genotypes that account for the 3 phenotypes (ie, extensive metabolizer, intermediate metabolizer, and poor metabolizer) had a significant impact on platelet reactivity with clopidogrel (174.9±54.0 vs. 193.1±56.5 vs. 240.6±25.4 PRU, P<0.001) but not prasugrel (147.0±51.9 vs. 147.5±58.3 vs. 184.4±38.3 PRU, P=0.15). CONCLUSIONS: Low-dose prasugrel achieves stronger platelet inhibition than clopidogrel in Japanese patients with stable CAD.

DOI： 10.1253/circj.CJ-15-0546

Pharmacogenetic-guided algorithms to estimate personalized dose or individual responses to anti-thrombotic drugs. Reviewed

Ariyoshi, N

Pers Med Univers. 4 13 - 22 2015

薬剤師と医師による定期処方協働入力の試みとその評価

山口洪樹, 三浦剛, 石井晃, 石田敬一, 鈴木貴明, 有吉範高, 松宮護郎, 石井伊都子

日本病院薬剤師会雑誌 50 ( 11 ) 1303 - 1307 2014.11

Language：Japanese Publisher：(一社)日本病院薬剤師会

千葉大学医学部附属病院において毎週1週間分処方される定期処方は、処方入力自体なされていない例(処方もれ)や経過中に変更された投与量が反映されていない例(処方ミス)が散見された。そこで、医療安全の確保を目的として、心臓血管外科病棟において薬剤師と医師が病棟で協議しながら定期処方を決定し入力する「協働入力」を開始した。取り組み開始後に処方もれはみられなくなり、処方ミスによる処方修正件数は開始前45件(22.4%)に対し開始後9件(3.4%)と有意な減少(p<0.01)がみられた。さらに、医師へのアンケート結果からも協働入力に対して好意的な回答が多く得られ、処方入力の際に患者の病態や服薬状況を理解している病棟薬剤師がその場で提案・情報提供できていることが、医師からの高い評価につながったと考えられた。以上により、本取り組みはチーム医療の推進につながり、医療安全および処方の適正化に貢献したと考えられた。(著者抄録)

Genetic and non-genetic factors responsible for antiplatelet effects of clopidogrel in Japanese patients undergoing coronary stent implantation: an algorithm to predict on-clopidogrel platelet reactivity. Reviewed International journal

Go Miura, Noritaka Ariyoshi, Yasunori Sato, Hiroki Yamaguchi, Yo Iwata, Yoshihide Fujimoto, Yoshio Kobayashi, Itsuko Ishii

Thrombosis research 134 ( 4 ) 877 - 83 2014.10

Language：English Publishing type：Research paper (scientific journal)

INTRODUCTION: Antiplatelet effects of clopidogrel appear to be affected by various factors including genetic polymorphism. So far, there has been little information about the response of clopidogrel in Asians, whose prevalence of a CYP2C19 loss-of-function (LOF) allele is high. METHODS AND RESULTS: We investigated background and clinical factors affecting on-clopidogrel platelet reactivity in Japanese patients undergoing coronary stent implantation (n=114). In univariate analysis, antiplatelet effects of clopidogrel in a steady state were associated with not only CYP2C19 genotypes but also several factors including dyslipidemia. In addition, we developed an algorithm that can estimate P2Y12 Reaction Units (PRU) in a steady state by multiple regression analysis and evaluated the adequacy of the algorithm by the Akaike Information Criterion. CONCLUSIONS: We revealed several factors influencing on-clopidogrel platelet reactivity in Japanese patients. We also succeeded in developing an algorithm that estimates PRU in a steady state, although it is uncertain whether the algorithm can be applied to other populations.

DOI： 10.1016/j.thromres.2014.07.018

バンコマイシン血中濃度コントロールに難渋した難治性腹水患者の1例 Reviewed

高塚博一, 山崎伸吾, 對田尚, 清野宗一郎, 竹田真理子, 鈴木達也, 関本匡, 丸山紀史, 鈴木貴明, 有吉範高, 横須賀收, 石井伊都子

TDM研究 31 ( 3 ) 166 - 166 2014.5

ボリコナゾールの胸腔内投与が血中濃度に与える影響 Reviewed

山崎伸吾, 渡邉哲, 高塚博一, 横山威一郎, 亀井克彦, 田川哲三, 溝渕輝明, 鈴木貴明, 有吉範高, 石井伊都子

日本化学療法学会雑誌 62 ( Suppl.A ) 284 - 284 2014.5

IMPACT OF OATP2B1 EXPRESSION ON CELL PROLIFERATION IN ER-POSITIVE BREAST CANCER Reviewed

Jun Matsumoto, Noritaka Ariyoshi, Masahiro Sakakibara, Takeo Nakanishi, Ikumi Tamai, Takeshi Nagashima, Itsuko Ishii

DRUG METABOLISM REVIEWS 45 268 - 268 2014.1

Decrease in the amount of lactose as a diluent for the powder dispensing and its evaluation

Mariko Tsukiji, Kazushi Masuda, Takaaki Suzuki, Noritaka Ariyoshi, Itsuko Ishii

Chiba Medical Journal 90 ( 6 ) 205 - 210 2014

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Chiba Medical Society

We have added pharmacologically inactive agent, lactose, to adjust total weight of one piece of packaged medicine to 0.3 g, when the quantity of medicine required are less than 0.3 g in the Pharmacy of Chiba University Hospital. However, it is sometimes difficult for children to ingest 0.3 g of powder. In addition, patients who need to intake several drugs simultaneously from a tube have some troubles in obstruction of the tube. On the other hand, variation in weight of packaged medicine is getting smaller due to progress of automatic powder packaging machine and the surrounding instruments. Thus we investigated whether we could decrease a total amount of packaged medicine with keeping variation of the weight within the range of specification. Weight variation of dispensed powder and loss of drug during dispensing or taking out the powder from packaging were within the range of specification even in the case of 0.2g of packaged powder, suggesting that we could decrease the total amount of powder to 0.2g without diminishing the quality. Moreover, we revealed that this change made us free from about one tenth of dilution works.

Functional characterization of seven single-nucleotide polymorphisms of the steroid sulfatase gene found in a Japanese population Reviewed

MATSUMOTO Jun, ARIYOSHI Noritaka, ISHII Itsuko, KITADA Mitsukazu

Journal of human genetics 58 ( 5 ) 267 - 272 2013.5

遺伝子検査と薬物治療

有吉範高

日本病院薬剤師会雑誌 49 ( 4 ) 351 - 357 2013.4

Language：Japanese Publisher：(一社)日本病院薬剤師会

分子生物学の進展によるヒト化技術の成功により多数の抗体医薬品が発売され、低分子の医薬品開発においても、癌の領域を中心に分子標的薬が次々に開発・上市されている。体細胞変異および遺伝子多型の両者が薬物応答性と関連していることが明らかになった今、これらの情報を無視して医薬品の適正使用を行うことは困難になりつつある。分子標的薬の投与適格性にかかわる遺伝子検出、遺伝子増幅確認、薬物応答性と関連する体細胞変異について述べた。

簡易遺伝子診断法の開発・実臨床への適用による医薬品適正使用の推進

有吉範高

医療薬学 39 ( 2 ) 61 - 76 2013.2

Language：Japanese Publisher：(一社)日本医療薬学会

薬物応答性の個体差をもたらす原因に関する研究は、分子レベルから遺伝子レベルへの時代に突入した。2000年から開始されたミレニアム・ゲノム・プロジェクトや、ファルマスニップコンソーシアムによる研究を皮切りに、今日まで多数の大型プロジェクトが実施されている。大型プロジェクトがもたらしたもの、研究成果が医療現場でなかなか活かせない理由、医療現場における遺伝子多型診断の実際、薬物応答性の個体差をもたらす多型以外の遺伝子情報、遺伝子診断を取り入れた臨床研究について述べた。

Other Link： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2013&ichushi_jid=J03520&link_issn=&doc_id=20130213430001&doc_link_id=10.5649%2Fjjphcs.39.61&url=https%3A%2F%2Fdoi.org%2F10.5649%2Fjjphcs.39.61&type=J-STAGE&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00007_3.gif

Development of a simple method for detection of the HLA-A*31:01 allele Reviewed

Kazuki Uchiyama, Fumika Kubota, Noritaka Ariyoshi, Jun Matsumoto, Itsuko Ishii, Mitsukazu Kitada

Drug Metab Pharmacokinet . 28 ( 5 ) 435 - 438 2013

眼科クリニカルパスにおける点眼手技評価方法の統一化に向けた取り組み

山口洪樹, 三浦剛, 舩本智津子, 加藤陽子, 加瀬千鶴, 石井晃, 仲佐啓詳, 中村裕義, 有吉範高, 北田光一

日本病院薬剤師会雑誌 48 ( 11 ) 1366 - 1370 2012.11

Language：Japanese Publisher：(一社)日本病院薬剤師会

千葉大学医学部附属病院では眼科白内障および硝子体手術のクリニカルパスに薬剤管理指導を導入し、薬剤師と看護師が協働して点眼指導を行ってきた。しかし、患者の点眼手技を評価する際に、評価者側に個人差があることが問題となっていた。新評価法では点眼手順に沿って6項目(手をふく、瞼をふく、目と点眼容器の距離、的外れ、滴下量、涙嚢部圧迫)評価を行うこととし、各項目の達成基準を設定した。達成した度合いに応じて各項目にA〜Dのスコアをつけ看護師と評価を共有した。新評価法導入後の評価を行った結果、評価者側の個人差は減少していることが示唆された。また、眼科看護師へのアンケート結果から新規評価法は抵抗感なく受け入れられており、運用面からも高い評価を得ていると考えられた。以上により、今回作成した点眼手技の新評価法でクリニカルパスにおける点眼指導の標準化と質的向上に貢献することができたと考えられる。(著者抄録)

FPIA法を用いた薬物モニタリングへの蛍光眼底造影剤の干渉 Reviewed

三浦剛, 中村裕義, 山形真一, 山崎伸吾, 仲佐啓詳, 有吉範高, 山本修一, 北田光一

TDM研究 29 ( 4 ) 83 - 88 2012.9

Language：Japanese Publisher：(一社)日本TDM学会

我々は、蛍光眼底造影検査(FAG)後の患者の蛍光偏光免疫測定(FPIA)法による血中シクロスポリン(CYA)濃度測定において、バックグラウンドの異常高値により測定不能となった症例を経験した。その原因として、FAGの際に投与された蛍光眼底造影剤フルオレセイン(F)による影響が疑われたため、FPIA法によるCYA、カルバマゼピン(CBZ)および酵素免疫測定法(MEIA法)によるジゴキシン(DIG)濃度測定に及ぼすFの影響について検討を行った。その結果、CYAでは全血液中F濃度が31.25ng/mLまで、CBZでは血清中F濃度は0.5μg/mLまでは薬物の測定値に影響することなく測定可能であったが、この濃度を超過すると測定不能となった。一方、DIGではF濃度が100μg/mLにおいても測定が可能であった。FPIA法による薬物血中濃度測定をFAG実施当日に行う場合には、FAG実施前に採血する必要があると考えられた。(著者抄録)

スナップ・ショットを用いた抗菌薬の適正使用推進の試み感染制御チームとの連携

三浦剛, 中村裕義, 千葉均, 井上智香子, 瀬川俊介, 渡辺正治, 渡辺哲, 石和田稔彦, 佐藤武幸, 仲佐啓詳, 有吉範高, 北田光一

日本病院薬剤師会雑誌 48 ( 8 ) 977 - 980 2012.8

Language：Japanese Publisher：(一社)日本病院薬剤師会

使用において特に注意を要する抗菌薬(以下、特定抗菌薬)の院内における適正使用の推進を目的として、感染制御チーム(以下、ICT)と連携して定点状況調査(以下、スナップ・ショット)を用いた使用状況の把握と病棟担当薬剤師による問題症例に対する介入を試みた。本取り組み開始前後1年間における特定抗菌薬の平均使用件数および14日以上の長期投与件数を比較したところ、いずれも開始後有意な減少が認められた。また、問題症例に対するICTの提言内容は、薬剤の選択と用法・用量の変更を要するものが半数以上を占め、これらの提言に対する遵守率は96%であった。以上、スナップ・ショットを用いた問題症例の抽出と、それに続く病棟担当薬剤師とICTの連携による指導等の取り組みにより、院内における特定抗菌薬の適正使用が推進されたと考えられた。(著者抄録)

使用実態からみたTPNの適正使用に関する検討

新井健一, 山本晃平, 竹田真理子, 鍋谷圭宏, 仲佐啓詳, 中村裕義, 有吉範高, 北田光一

日本病院薬剤師会雑誌 48 ( 7 ) 853 - 856 2012.7

Language：Japanese Publisher：(一社)日本病院薬剤師会

中心静脈カテーテルを用いた高カロリー輸液(以下、TPN)は、経腸栄養療法に比べ、その使用頻度は減少傾向であるが、消化管を使用できない患者に対しては必要な栄養療法である。本研究では、千葉大学医学部附属病院におけるTPNの使用実態を調査し、問題点を具体化し、適正使用にいかに寄与すべきかを検討した。本調査より、TPN投与期間の短い症例、糖質の投与速度超過、脂肪乳剤の併用不足、投与熱量不足などの実態が明らかとなった。TPNを有益な栄養療法とするために、TPN投与における当該患者の適応症の確認、投与速度、必要熱量を体重より算出し、指示が適切でない場合は、適正な情報を提供することが必要であると考えられた。本結果を基に今後医師への医薬品情報(DI)提供、薬剤管理指導、栄養サポートチーム(NST)による院内講習会などを通してTPNの適正使用を推進する必要があると考えられた。(著者抄録)

シクロスポリン細粒の分包調剤後の光安定性に関する検討 Reviewed

山崎伸吾, 中村裕義, 山形真一, 仲佐啓詳, 有吉範高, 北田光一

医薬品情報学 14 ( 1 ) 35 - 39 2012.5

Language：Japanese Publisher：(一社)日本医薬品情報学会

シクロスポリン(CsA)細粒の分包調剤後の光安定性について検討した。700ルクスの白色蛍光灯による光照射を1日12時間行う条件下において3種の保存形態、分包紙のみで被覆のない状態(曝光保存)、外面がパルプで内面をポリエチレンコーティングされた薬袋内に保管(薬袋内保存)、遮光ポリエチレン袋に保管(遮光袋内保存)で実施した。薬袋内あるいは遮光袋内で保管した場合には3ヵ月までは安定であったが、曝光群においては同様の条件下でCsAの外観が白色に変化し、12週後においてわずかながら規格値である残存率(95%)を下回った。3種の保管環境での溶出パターンの変化について検討し、残存率の低下を反映した溶出率の低下は認めたが、溶出パターンには変化は認めなかった。

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Correlation between antizyme 1 and differentiation of vascular smooth muscle cells cultured in honeycomb-like type-I collagen matrix Reviewed

Itsuko Ishii, Takaaki Suzuki, Hiromi Kaneko, Masashi Uchida, Yukari Suzuki, Kyohei Higashi, Satoko Yagi, Noritaka Ariyoshi, Kazuei Igarashi, Mitsukazu Kitada

Amino Acids 42 ( 2-3 ) 565 - 575 2012.2

Language：English Publishing type：Research paper (scientific journal)

Vascular smooth muscle cells (SMC) are able to proliferate when cultured on plates, but become differentiated when maintained in three-dimensional type I collagen matrices (honeycombs). SMC grown in honeycombs contained a low level of polyamines due to the presence of antizyme 1 (AZ1), a negative regulator of ornithine decarboxylase (ODC) and of polyamine uptake. To clarify the role of AZ1 in differentiation of SMC in honeycombs, an ODC gene was stably transfected into SMC (ODCSMC). Although proliferation of ODC-SMC on plates was accelerated together with an increase in phosphorylated focal adhesion kinase (FAK) and a decrease in a-actin and myosin, maker proteins of differentiation, growth of ODCSMC ceased in honeycombs similarly to normal SMC with a low level of phosphorylated FAK and a high level of a-actin and myosin. AZ1 expression in ODC-SMC on plates was low, but that in honeycombs was high. Antizyme in ODC-SMC in honeycombs not only decreased the level of ODC but also inhibited polyamine uptake activity. These results taken together suggest that low levels of polyamines caused by AZ1 in SMC in honeycombs inhibit phosphorylation of FAK and enhance expression of a-actin and myosin, resulting in differentiation through inhibition of focal adhesions. © Springer-Verlag 2011.

DOI： 10.1007/s00726-011-1034-8

Q172H Replacement Overcomes Effects on the Metabolism of Cyclophosphamide and Efavirenz Caused by CYP2B6 Variant with Arg262 Reviewed

Noritaka Ariyoshi, Miyuki Ohara, Mayumi Kaneko, Sakino Afuso, Takuya Kumamoto, Hiroyoshi Nakamura, Itsuko Ishii, Tsutomu Ishikawa, Mitsukazu Kitada

DRUG METABOLISM AND DISPOSITION 39 ( 11 ) 2045 - 2048 2011.11

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS

There are a number of reports indicating that CYP2B6*6 (c.516G > T and c.785A > G) is responsible for decreased clearance of efavirenz (EFV), although increased disposition of cyclophosphamide (CPA) in individuals with this polymorphism was observed. Thus, we hypothesized that the effects of the two single nucleotide polymorphisms (SNPs) of CYP2B6*6 on the metabolism of drugs might be considerably different between these two agents. To clarify this possibility, we expressed two major variants of this enzyme, CYP2B6.6 (Q172H and K262R) and CYP2B6.4 (K262R), and investigated metabolic activities of these variants toward EFV and CPA. Kinetic analyses clearly indicated that CYP2B6.4 possessed enhanced metabolic activity toward EFV compared with that of the wild-type enzyme (CYP2B6.1), whereas CPA was metabolized less efficiently by CYP2B6.4 than by CYP2B6.1. On the other hand, CYP2B6.6 showed a completely opposite character, suggesting that Q172H gives inverse effects on metabolic activities of CYP2B6 affected by K262R. Although it is recognized that effects of amino acid change in cytochrome P450 on the metabolic activity depend on substrates, this study revealed SNPs giving an opposite effect on the metabolism of two clinically important drugs currently used. Furthermore, this study provides the first evidence that Q172H can reverse the direction of the effect caused by K262R in CYP2B6 on the metabolism of certain drugs.

DOI： 10.1124/dmd.111.039586

Degradation of filamin induces contraction of vascular smooth muscle cells in type-I collagen matrix honeycombs Reviewed

Masashi Uchida, Itsuko Ishii, Kaori Hirata, Fumiko Yamamoto, Kaori Tashiro, Takayoshi Suzuki, Yuji Nakayama, Noritaka Ariyoshi, Mitsukazu Kitada

Cellular Physiology and Biochemistry 27 ( 6 ) 669 - 680 2011

Language：English Publishing type：Research paper (scientific journal)

Background: Dedifferentiated rabbit vascular smooth muscle cells (SMCs) exhibit similar features to differentiated SMCs when cultured in three-dimensional matrices of type-I collagen called "honeycombs," but the mechanism is unknown. The role of filamin, an actin-binding protein that links actin filaments in SMCs, was investigated. Methods: Filamin and other related proteins were detected by western blot analysis and immunofluorescence staining. Honeycomb size was measured to confirm the contraction of SMCs. Results: Full-length filamin was expressed in subconfluent SMCs cultured on plates
however, degradation of filamin, which might be regulated by calpain, was observed in confluent SMCs cultured on plates and in honeycombs. While filamin was co-localized with β-actin in subconfluent SMCs grown on plates, filamin was detected in the cytoplasm in SMCs cultured in honeycombs, and degraded filamin was mainly detected in the cytoplasmic fraction of these cells. In addition, β-actin expression was low in the cytoskeletal fraction of SMCs cultured in honeycombs compared with cells cultured on plates, and the size of the honeycombs used for culturing SMCs was significantly reduced. Conclusion: These data suggest that degradation of filamin in SMCs cultured in honeycombs induces structural weakness of β-non-muscle actin filaments, thereby permitting SMCs in honeycombs to achieve contractility. Copyright © 2011 S. Karger AG, Basel.

DOI： 10.1159/000330076

薬剤師の観点から見たNutrition Support Team活動の現状と問題点

新井健一, 山本晃平, 鍋谷圭宏, 櫻井健一, 古川勝規, 野本尚子, 仲佐啓詳, 中村裕義, 有吉範高, 北田光一

日本病院薬剤師会雑誌 46 ( 12 ) 1657 - 1660 2010.12

Language：Japanese Publisher：(一社)日本病院薬剤師会

千葉大学医学部附属病院では2006年4月よりnutrition support team(以下、NST)活動を開始した。今回我々は、これまでに介入した症例を基に、薬剤師の観点から活動の問題点と今後の課題を検討した。NSTへの依頼内容は、栄養状態の改善と食欲不振対策が全体の60%以上を占め、その患者の多くは悪性腫瘍であった。悪性腫瘍である患者の多くは病態により栄養状態が悪化しており、またがん化学療法を施行している場合には嘔気・嘔吐などの副作用により食欲不振に陥るため介入依頼となっていた。NSTが介入するも、血清アルブミン値(血清ALB値)が低下した症例について検討した結果、がん化学療法による栄養状態悪化が最も多かった。がん化学療法による嘔気・嘔吐対策、食事・経腸栄養剤との相互作用対策など医薬品に関する事例が介入内容に多く認められたため、NSTに属する薬剤師として医薬品の適正使用を推進し、患者の栄養改善に向けた取り組みが必要と考えられた。(著者抄録)

医薬品の服用時期および最大投与量の設定根拠を迅速に検索できるデータベースの構築と疑義照会への活用

新井さやか, 大久保正人, 石島彩子, 長谷川敦, 仲佐啓詳, 中村裕義, 有吉範高, 北田光一

医薬品情報学 12 ( 2 ) 69 - 76 2010.11

Language：Japanese Publisher：(一社)日本医薬品情報学会

添付文書に記載された服用時期および最大投与量の設定根拠となる情報を迅速に検索可能なデータベース(DB)を構築し、その有用性について評価した。採用の内服薬(723品目)のうち、服用時期として、「起床時」「食前」「食直前」「食直後」「食間」「空腹時」の規定のある医薬品38品目、および最大投与量の規定のある184品目を対象とした。服用時期および最大投与量ともに、日常業務において比較的容易に検索可能である添付文書やIFから得られた割合は少なく、50%以上の医薬品で製薬企業への問い合わせが必要であった。設定根拠が不明な医薬品も服用時期で3品目、最大投与量で25品目存在した。服用時・期では二重盲検比較試験の結果に基づいて設定されている医薬品が38品目中17品目ある一方で、販売開始後に出版された雑誌の総説や症例報告などを設定根拠としている医薬品も存在した。稼動当初のDBのレコード数は313件であった。

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Kefiran reduces atherosclerosis in rabbits fed a high cholesterol diet. Reviewed

Masashi Uchida, Itsuko Ishii, Chika Inoue, Yoshie Akisato, Kenta Watanabe, Saori Hosoyama, Toshihiko Toida, Noritaka Ariyoshi, Mitsukazu Kitada

Journal of atherosclerosis and thrombosis 17 ( 9 ) 980 - 8 2010.9

Language：English Publishing type：Research paper (scientific journal)

AIM: Kefiran is an exopolysaccharide produced by Lactobacillus kefiranofaciens, and has been proposed to have many health-promoting properties. We investigated the antiatherogenic effect of kefiran on rabbits fed a high-cholesterol diet. METHODS: Male New Zealand White rabbits were fed a 0.5% cholesterol diet without (control group, n = 7) or with kefiran (kefiran group, n = 8) for eight weeks. The aorta was analyzed by histochemistry and atherosclerotic lesions were quantified. Lipids and sugars in serum were measured. Foam cell formation of RAW264.7 by βVLDL derived from both groups of rabbits was also investigated. RESULTS: Cholesterol, triglyceride and phospholipids levels of serum and lipoprotein fractions were not significantly different between these groups. Atherosclerotic lesions of the aorta in the kefiran group were statistically lower than those of the control group, with marked differences in the abdominal aorta. T-lymphocytes were not detectable in the aorta of the kefiran group. Cholesterol contents in stools were almost identical in both groups. Cholesterol content in the liver of the kefiran group was statistically lower than in the control group. Galactose content of βVLDL derived from the kefiran group was higher, and the lipid peroxidation level was much lower than in the control group. RAW264.7 macrophages treated with βVLDL from the kefiran group showed a more spherical shape and accumulated statistically lower cholesterol than macrophages treated with βVLDL from the control group. CONCLUSION: Orally derived kefiran is absorbed in the blood. Kefiran prevents the onset and development of atherosclerosis in hypercholesterolemic rabbits by anti-inflammatory and anti-oxidant actions.

Application of Akaike information criterion to evaluate warfarin dosing algorithm. Reviewed International journal

Takumi Harada, Noritaka Ariyoshi, Hitoshi Shimura, Yasunori Sato, Iichiro Yokoyama, Kaori Takahashi, Shin-ichi Yamagata, Mizuho Imamaki, Yoshio Kobayashi, Itsuko Ishii, Masaru Miyazaki, Mitsukazu Kitada

Thrombosis research 126 ( 3 ) 183 - 90 2010.9

Language：English Publishing type：Research paper (scientific journal)

INTRODUCTION: Several factors responsible for inter-individual differences in response to warfarin have been confirmed; however, unidentified factors appear to remain. The purpose of this study was to examine a simple method to evaluate whether optional variables are appropriate as factors to improve dosing algorithms. MATERIALS AND METHODS: All patients were Japanese. Genotyping of selected genes was conducted, and other information was obtained from medical record. Dosing algorithms were constructed by multivariate linear regression analyses and were evaluated by the Akaike Information Criterion (AIC). RESULTS AND CONCLUSIONS: Multivariate analysis showed that white blood-cell count (WBC), concomitant use of allopurinol, and CYP4F2 genotype are apparently involved in warfarin dose variation, in addition to well-known factors, such as age and VKORC1 genotype. We evaluated the adequacy of these variables as factors to improve the dosing algorithm using the AIC. Addition of WBC, allopurinol administration and CYP4F2 genotype to the basal algorithm resulted in decreased AIC, suggesting that these factor candidates may contribute to improving the prediction of warfarin maintenance dose. This study is the first to evaluate the warfarin dosing algorithm by AIC. To further improve the dosing algorithm, AIC may be a simple and useful tool to evaluate both the model itself and factors to be incorporated into the algorithm.

DOI： 10.1016/j.thromres.2010.05.016

Super-acute onset of tumor lysis syndrome accompanied by hypercytokinemia during treatment of Hodgkin's lymphoma with ABVD chemotherapy. Reviewed International journal

Takaaki Suzuki, Masahiro Takeuchi, Hiromi Saeki, Shingo Yamazaki, Hitomi Koga, Daijiro Abe, Miki Nishimura, Chiaki Nakaseko, Hiromitsu Nakasa, Hiroyoshi Nakamura, Noritaka Ariyoshi, Mitsukazu Kitada

Clinical therapeutics 32 ( 3 ) 527 - 31 2010.3

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Tumor lysis syndrome (TLS) is a group of life-threatening metabolic complications that can occur after initiation of cancer chemotherapy. Onset of TLS in the middle of chemotherapy, however, has not been reported previously in patients with hematologic malignancies. OBJECTIVE: We report a case of a patient who experienced TLS of super-acute onset accompanied by hypercytokinemia during chemotherapy treatment with a combination of doxorubicin (Adriamycin), bleomycin, vinblastine, and dacarbazine (ABVD). CASE SUMMARY: A 36-year-old Japanese man (height, 182 cm; weight, 83 kg; body surface area, 2.04 m(2)) was admitted to the hospital for the treatment of malignant lymphoma (clinical stage IVB Hodgkin's lymphoma). Chemotherapy was initiated using the ABVD regimen (doxorubicin [Adriamycin] 25 mg/m(2) by 30-minute infusion, bleomycin 9 mg/m(2) by 30-minute infusion, vinblastine 6 mg/m(2) by bolus injection, and dacarbazine 375 mg/m(2) by 2-hour infusion). During the dacarbazine infusion, the patient's body temperature rose from 36.5 degrees C to 42 degrees C; he experienced a convulsion and then lost consciousness. The convulsion was not suppressed despite the use of diazepam (5 mg IV twice) and phenytoin (500 mg IV). The patient was then transferred to the intensive care unit and sedated using a continuous infusion of midazolam (10 mg/h). Levels of serum lactate dehydrogenase, aspartate aminotransferase, uric acid, blood urea nitrogen, and creatinine evaluated shortly after the ABVD regimen were outside normal limits. In addition, interleukin-6 (IL-6) concentrations were elevated to 54,220 pg/mL. Continuous hemodiafiltration was immediately performed to lower the elevated levels of IL-6. The next day, IL-6 concentrations decreased to 97 pg/mL, and the patient was weaned from ventilator support and sedation. The patient had no adverse effects after the event. According to the results of an assessment using the Naranjo adverse drug reaction probability scale (score = 3), the development of TLS in this patient was possibly related to the chemotherapy regimen. CONCLUSIONS: ABVD chemotherapy was possibly associated with the super-acute onset of TLS in this patient. In addition, hypercytokinemia occurred with TLS, which led to pyrexia, convulsion, and loss of consciousness.

DOI： 10.1016/j.clinthera.2010.03.010

Six novel single nucleotide polymorphisms of the steroid sulfatase gene in a Japanese population. Reviewed International journal

Jun Matsumoto, Noritaka Ariyoshi, Itsuko Ishii, Mitsukazu Kitada

Drug metabolism and pharmacokinetics 25 ( 4 ) 403 - 7 2010

Language：English Publishing type：Research paper (scientific journal)

Steroid sulfatase (STS) is a microsomal enzyme responsible for the formation of 3beta-hydroxysteroid from the corresponding sulfate conjugates. Screening of all exons, exon-intron boundaries and the 5'-flanking region of the STS gene in 93 healthy Japanese individuals was carried out. Among seven single nucleotide polymorphisms (SNPs) identified in this study, six were novel, including one in the untranslated region of exon 1, one in exon 10, and four in the 5'-flanking region. The nonsynonymous SNP (1647G>A) in exon 10 caused amino-acid replacement, Val476Met, with a frequency of 0.014. The allele frequencies of the other SNPs were 0.071 for 155G>A, 0.007 for -21G>A, 0.014 for -1117T>C, 0.106 for -1588G>A, 0.007 for -2427G>A and 0.007 for -2837T>C.

Enhanced susceptibility of HLA-mediated ticlopidine-induced idiosyncratic hepatotoxicity by CYP2B6 polymorphism in Japanese. Reviewed International journal

Noritaka Ariyoshi, Yukako Iga, Koji Hirata, Yasunori Sato, Go Miura, Itsuko Ishii, Seiji Nagamori, Mitsukazu Kitada

Drug metabolism and pharmacokinetics 25 ( 3 ) 298 - 306 2010

Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)

Hepatotoxicity is the most frequent adverse drug reaction (ADR) in Japanese treated with ticlopidine (TP). We investigated the relationship between CYP2B6 haplotype and incidence of TP-induced hepatotoxicity in 114 Japanese patients. Although 4 haplotypes (*1A, *1H, *1J and *6B) accounted for more than 80% of the inferred haplotypes in both control (n=81) and case (n=22) subjects, the prevalence was apparently different: control, *1A>*6B>*1H>*1J and case, *1J>*1H>*1A>*6B. The reporter gene assay for the two SNPs, which comprise the *1H or *1J haplotype, suggested that the *1H and *1J haplotypes may be associated with the increased expression of CYP2B6, probably due to g.-2320T>C. Combination analysis of CYP2B6 and human leukocyte antigen (HLA) haplotypes revealed that individuals possessing CYP2B6*1H or *1J with HLA-A*3303 have the highest susceptibility to TP-induced hepatotoxicity (odds ratio, 38.82; 95%CI, 8.08-196.0, P<0.001). Although this is a preliminary case-control study with some limitations, it is the first example that HLA-induced idiosyncratic ADR may be modified by individual variation in CYP activities.

Unexpected serum level of vancomycin after oral administration in a patient with severe colitis and renal insufficiency. Reviewed International journal

S Yamazaki, H Nakamura, S Yamagata, G Miura, N Hattori, K Shinozaki, T Sadahiro, A Toyoda, H Nakasa, N Ariyoshi, S Oda, K Harigaya, M Kitada

International journal of clinical pharmacology and therapeutics 47 ( 11 ) 701 - 6 2009.11

Language：English Publishing type：Research paper (scientific journal)

OBJECTIVE: To report a case in which the serum concentration of vancomycin (VCM) reached the supratherapeutic range following oral administration in a patient with severe pseudomembranous colitis and renal insufficiency. CASE SUMMARY: A 65-year-old, 70 kg weighing man with severe acute pancreatitis and acute renal failure was subjected to continuous hemodiafiltration (CHDF). CHDF could only be performed intermittently because of the unstable circulation dynamic of this patient. After admission, intravenous VCM therapy was initiated. Thereafter, oral VCM administration was begun (0.5 g every 6 h). Despite the discontinuation of intravenous VCM after the first 2 days of oral VCM, the serum VCM concentration increased gradually to 49.8 mg/l over a period of 2 weeks from the initiation of oral administration (34.4 mg/l). Based on pharmacokinetic analysis, the bioavailability of VCM was estimated to over 33%. Autopsy findings indicated broadly distributed necrosis on the lamina propria of the mucosa throughout all parts of the intestine below the duodenum. DISCUSSION: This case indicates necessity of the careful monitoring after oral high-dose VCM administration in a patient with a broadly distributed necrosis and renal insufficiency. CONCLUSIONS: TDM should be considered according to renal function, the severity of enteritis and the total dosage of oral VCM administration.

ワルファリン治療個別化に利用可能な簡易・同時遺伝子診断法の確立

有吉範高, 中澤一純, 北田光一

医療薬学 35 ( 8 ) 551 - 557 2009.8

Language：Japanese Publisher：(一社)日本医療薬学会

ワルファリン(Wf)による抗凝固療法はその投与量に個体差が大きく、Wf治療個別化及び医療現場におけるルーチンな遺伝子診断への応用を視野に入れ、遺伝子診断が必須となることが想定されるVKORC1とCYP2C9の簡便な同時遺伝子多型診断法の確立を試みた。その結果、本研究では採血することなく、低侵襲な方法で良質なDNAを得て、その試料で実施できる遺伝子診断を行う新規多型診断法を確立した。本遺伝子診断法ではコスト面や迅速性に問題のあった従来法に比べ、1回のPCRと電気泳動で同時にVKORC1とCYP2C9の遺伝子型の判定結果を正確・簡便・低コストで迅速に得ることが可能である。しかし、Wf感受性は遺伝的要因のみならず患者の生理的要因や食事・併用薬物などの環境要因によっても著しい影響を受けるため、VKORC1やCYP2C9の遺伝子多型情報だけでは患者の維持投与量の正確な事前予測は困難であり、今後さらなる研究が必要と考えられた。

Other Link： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2009&ichushi_jid=J03520&link_issn=&doc_id=20090813400004&doc_link_id=10.5649%2Fjjphcs.35.551&url=https%3A%2F%2Fdoi.org%2F10.5649%2Fjjphcs.35.551&type=J-STAGE&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00007_3.gif

上皮成長因子受容体(EGFR)体細胞変異の簡易遺伝子診断法開発と臨床応用

有吉範高, 伊豫田明, 廣島健三, 渋谷潔, 滝口裕一, 中谷行雄, 北田光一

医療薬学 35 ( 7 ) 468 - 477 2009.7

Language：Japanese Publisher：(一社)日本医療薬学会

比較的迅速で、コストが安く、感度も満足しうる簡便な上皮成長因子受容体(EGFR)遺伝子変異診断法を開発した。構築した診断法は欠失領域と相補的なプライマーが結合できなくなるため、そのいずれの欠失パターンであっても診断可能である。更にエクソン21のL861Qは、その頻度はL858Rに比べ1/10以下と少ないが、L858R診断のために増幅したPCR産物を別の制限酵素で切断するだけで判定可能である。しかも両エクソン由来の増幅断片は同じPCR条件下で増幅できるように設計してあるため、一度のPCRで3種類の変異箇所の診断に必要な増幅産物を全て知ることができる。PCRのサイクル数は試料から回収されるDNA量によって多少変更が必要であるが十分な品質と量のDNAが得られる試料の場合は30サイクル、FFPE薄切標本を試料とした場合は35〜40サイクルで十分であった。

Other Link： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2009&ichushi_jid=J03520&link_issn=&doc_id=20090714420004&doc_link_id=%2Fdb5pharm%2F2009%2F003507%2F004%2F0468-0477%26dl%3D0&url=https%3A%2F%2Fwww.medicalonline.jp%2Fjamas.php%3FGoodsID%3D%2Fdb5pharm%2F2009%2F003507%2F004%2F0468-0477%26dl%3D0&type=MedicalOnline&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00004_2.gif

Growth inhibition and differentiation of cultured smooth muscle cells depend on cellular crossbridges across the tubular lumen of type I collagen matrix honeycombs. Reviewed International journal

Takaaki Suzuki, Itsuko Ishii, Akira Kotani, Michi Masuda, Kaori Hirata, Madoka Ueda, Takahiro Ogata, Takanori Sakai, Noritaka Ariyoshi, Mitsukazu Kitada

Microvascular research 77 ( 2 ) 143 - 9 2009.3

Language：English Publishing type：Research paper (scientific journal)

Although rabbit vascular smooth muscle cells (SMCs) showed a differentiated phenotype in three-dimensional type I collagen matrices (honeycombs, diameter of pores=200-500 microm), mouse vascular SMCs proliferated in honeycombs having the same pore size. Here we investigated the relationship between pore sizes of honeycombs and differentiation of SMCs using various pore sizes of honeycombs. Rabbit SMCs (length: 200+/-32 microm) and mouse SMCs (49+/-10 microm) formed crossbridges in honeycombs with 200-300 microm and less than 200 microm of pores, respectively. Both SMCs spread on the inner wall but did not form crossbridges in honeycombs with larger pores. [(3)H]Thymidine incorporation and cell number of both SMCs were decreased when the crossbridges were formed in honeycombs. Because proliferation inhibition and crossbridge formation were observed in the culture of rabbit and mouse SMCs using 200-300 microm and less than 200 microm pore sized honeycombs, respectively, these data suggested that forming crossbridges was important for the inhibition of proliferation of SMCs. Rabbit SMCs differentiation was accompanied by the expression of caldesmon heavy chain when cultured in honeycombs having less than 300 microm pores. Proliferation of mouse SMCs stopped in honeycombs having less than 200 microm pores, but caldesmon heavy chain was not detected despite the expression of its mRNA. Proliferation of SMCs stopped on plates when cells reached confluent state, however, caldesmon heavy chain was not expressed. These data suggested that an appropriate structure and suitable honeycomb pore size are important for the differentiation of SMCs.

DOI： 10.1016/j.mvr.2008.08.006

粉砕調剤後の安定性の情報に関する調査・検討

増田和司, 今関美智子, 高橋香, 大久保正人, 長谷川敦, 中村貴子, 中澤一純, 仲佐啓詳, 中村裕義, 有吉範高, 北田光一

医薬品情報学 10 ( 4 ) 291 - 297 2009.3

Language：Japanese Publisher：(一社)日本医薬品情報学会

錠剤の粉砕後の安定性に関して、現場の薬剤師が得られる情報について調査し、一定の基準を設定し粉砕後の保存可能期間や保存時の注意事項について検討し、粉砕調剤の可否を検討する上での問題点について考察した。粉砕調剤の可否を判断する上で製剤学的に問題がある医薬品(徐放性製剤、腸溶性製剤、舌下錠)や粉砕調剤の必要性が低いと考えられる医薬品(速崩壊性製剤など)を除外した364成分406品目を対象とした。製薬企業からの情報提供や文献検索により錠剤の粉砕後の安定性試験の情報を得ることができた医薬品は、257成分269品目であった。このうち226品目の粉砕後の安定性情報は、製薬企業の有する非公表の情報からのみ入手可能であった。安定性を評価するうえで最低限必要と考えられる性状と有効成分含量の両項目が検討されていた医薬品は248成分259品目であった。

病院薬剤部における調剤インシデントレポート内容の分析に基づいた対策とその効果

高橋香, 中澤一純, 長谷川敦, 中村貴子, 中村裕義, 有吉範高, 北田光一

日本病院薬剤師会雑誌 44 ( 12 ) 1761 - 1764 2008.12

Language：Japanese Publisher：(一社)日本病院薬剤師会

千葉大学医学部附属病院薬剤部調剤室では、調剤薬の最終鑑査で発見されたレベル0のインシデント事例について収集を行ってきた。今回その事例を分析し、調剤ミスを回避するための対策を実施することで、ミスの発生がどのように変化したか調査した。平成14,15および17年度においてレベル0のインシデント事例を基に採用薬の見直し、医薬品の配置変更や処方せんへの印字工夫などの対策を行った。その結果、調剤ミス発生率は経年的に低下し、診療科別でもほとんどの診療科で低下傾向を認めた。また曜日別では、どの年度においても月曜日に高く水曜日に低い傾向を示したが、平成17年度はすべての曜日において低値であった。以上のことより、レベル0のインシデント事例を収集・分析し対策を講じることにより、調剤ミスをより一層減少させることが可能であることが示された。(著者抄録)

レトロスペクティブ調査からAnti-Methiciliin-Resistant Staphylococcus aureus Agentsの適正使用を考える起炎菌・検査値・therapeutic drug monitoringを用いた薬剤使用前後における検討

中村安孝, 横山威一郎, 橋本典子, 長谷川敦, 仲佐啓詳, 中村裕義, 渡辺正治, 野村文夫, 渡邊哲, 猪狩英俊, 佐藤武幸, 中澤一純, 有吉範高, 北田光一

薬学雑誌 128 ( 7 ) 1073 - 1079 2008.7

Language：Japanese Publisher：(公社)日本薬学会

千葉大学医学部附属病院における抗メチシリン抵抗性S.aureus剤(抗MRSA剤)の2001〜2005年迄の5年間の使用記録から、4日以上使用した症例を集めた。それぞれの症例について抗MRSA剤投与前後の細菌検査、臨床検査データとTDMが行われたかどうかを調べた。抗MRSA剤のうちで硫酸アルベカシンを除いて、塩酸バンコマイシンとテイコプラニンを投与した患者数と薬剤量が年ごとに増加する傾向が認められた。抗MRSA剤投与によって体温、C反応性蛋白質、白血球数が有意に低下した。抗MRSA剤投与患者の75.6%で細菌検査が行われ、そのうち72.4%でMRSAが検出された。TDMが行われた患者の割合は60%で、他の検査に比べて僅かしか検査されていなかった。抗MRSA薬がMRSA以外の起炎菌にも使用され、一部治療終了後も保菌状態であることが確認された。TDMにより適正な治療域へ早期に導く必要性があると推察された。

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Probable interaction between warfarin and antitumor agents used in R-ESHAP chemotherapy. Reviewed International journal

Takaaki Suzuki, Hitomi Koga, Shingo Yamazaki, Hiromi Saeki, Hiroaki Tanaka, Miki Nishimura, Chiaki Nakaseko, Hiromitsu Nakasa, Hiroyoshi Nakamura, Noritaka Ariyoshi, Mitsukazu Kitada

Clinical therapeutics 30 ( 6 ) 1155 - 9 2008.6

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: The pharmacologic effects of warfarin might be altered by various factors, including drug-drug interaction. CASE SUMMARY: A 49-year-old Japanese man (height, 174 cm; weight, 68 kg) presented with a 20-month history of malignant lymphoma (diffuse large B cell lymphoma, clinical stage IV). He was treated with a combination of rituximab chemotherapy and etoposide, cisplatin, high-dose cytarabine, and methyl-prednisolone (R-ESHAP). He had been receiving warfarin for the secondary prevention of pulmonary embolism with deep venous thrombosis. When R-ESHAP was started, international normalized ratio (INR) increased from 1 to 5. This phenomenon was observed again in the second R-ESHAP. The INR was increased from 2.44 to 4.71 during chemotherapy but was returned to within the normal range (1.05; normal range: 0.81-1.009) 5 days after chemotherapy was completed. CONCLUSION: In this patient, R-ESHAP chemotherapy might have affected warfarin anticoagulation sensitivity; thus, careful monitoring of INR is essential, particularly in patients receiving warfarin who undergo R-ESHAP chemotherapy.

DOI： 10.1016/j.clinthera.2008.06.008

当院における術後感染予防薬の使用状況と本邦ガイドラインとの比較検討

橋本典子, 中村安孝, 仲佐啓詳, 中村裕義, 有吉範高, 北田光一

医薬品情報学 10 ( 1 ) 48 - 52 2008.5

Language：Japanese Publisher：(一社)日本医薬品情報学会

当院における術後感染予防のための抗菌薬使用状況を調査し、日本感染症学会「抗菌薬使用のガイドライン」(GL)と比較した。対象期間の手術5111件のうち、4060件で抗菌薬が使用され、第一世代セフェム系が32.4%と最も多かった。使用状況の評価で、薬剤種類については70.2%がGLの推奨範囲内であったが、使用期間は23.5%が推奨範囲内であった。手術の種類別にみると、使用薬剤・期間ともGLの推奨範囲内であった割合が高かったのは乳腺・甲状腺外科97.9%、食道・胃腸外科の胃切除術83.9%であった。一方、使用薬剤が推奨範囲外であった割合が50%以上は食道・胃腸外科(食道手術)と呼吸器外科(胸腔鏡下・開胸手術)で、食道手術では推奨よりもスペクトルの広いフロモキセフが、呼吸器外科では推奨にないアンピシリン/スルバクタムが使用されていた。使用期間については眼科、心臓血管外科、循環器内科、周産期・母性科、呼吸器外科(胸腔鏡下・開胸手術)で推奨範囲外が多かった。

Population pharmacokinetics of oral busulfan in young Japanese children before hematopoietic stem cell transplantation. Reviewed International journal

Hiroyoshi Nakamura, Takeyuki Sato, Kenji Okada, Go Miura, Noritaka Ariyoshi, Kazuyoshi Nakazawa, Mitsukazu Kitada

Therapeutic drug monitoring 30 ( 1 ) 75 - 83 2008.2

Language：English Publishing type：Research paper (scientific journal)

The objectives of this study were to develop a population pharmacokinetic model and to determine the covariates affecting the pharmacokinetics of busulfan in Japanese pediatric patients who received high-dose oral busulfan as a conditioning regimen before hematopoietic stem cell transplantation. Population analysis was performed using retrospective therapeutic drug monitoring data (including test dose data) from 103 children. Their ages ranged from 2 months to 11 years old (mean age, 30 months; median age, 18 months). The plasma concentration of busulfan in all 1028 samples was measured with the same high-performance liquid chromatography method. Maximum likelihood estimates were sought for pharmacokinetic parameters with the NONMEM program. The best structural covariate-free model for busulfan was a one-compartment model with an exponential error model to account for intersubject variability and a proportional error model to account for intrasubject variability. The apparent oral clearance was found to be correlated with age, aspartate transaminase, and type of disease (malignant disease or other). The apparent volume of distribution was related to body weight. The busulfan formulation (1% powder form or crystal form) and dose (milligrams per kilogram) influenced the absorption rate constant. It was estimated that oral clearance expressed per kilogram of body weight is low at early infancy, then increases to a maximum at approximately 2 years of age and, thereafter, decreases. In conclusion, we have developed a population pharmacokinetic model of oral busulfan in children, particularly for those younger than 4 years old, that takes into consideration not only body size, but also several other covariates.

DOI： 10.1097/FTD.0b013e3181621cde

医薬品数量管理システム構築と医薬品購入費および業務効率に対する効果

大久保正人, 増田和司, 長谷川敦, 中村貴子, 野口昇, 中村裕義, 中澤一純, 有吉範高, 北田光一

日本病院薬剤師会雑誌 44 ( 2 ) 233 - 236 2008.2

Language：Japanese Publisher：(一社)日本病院薬剤師会

医薬分業の進展により院内で調剤される医薬品数量は減少したが、投与日数制限が原則撤廃されたために一度に処方される数量は増加しており、調剤室に置く医薬品数量の調整が新たな課題となっている。そこで、本研究では調剤室の医薬品数量の適正化を目的として、病院情報システムのデータを利用した医薬品請求システムの構築を試みた。まず、処方オーダリングシステムのデータから対象医薬品の処方量を医薬品別・月別に集計し、適正な一回請求量の設定と不動医薬品の選別化を行った。次に、処方が低頻度な高額医薬品の管理のために、患者の次回来院予定日を電子カルテから検索するシステムを構築した。これらのシステム稼動前後で、年度末棚卸金額は40.4%減少、特に外用剤では54.8%と著しく減少した。一方で、臨時発注件数には顕著な増加は認められず、今回の取り組みが在庫適正化に寄与したと考えられた。(著者抄録)

Relationship between Kaposi's varicelliform eruption in Japanese patients with atopic dermatitis treated with tacrolimus ointment and genetic polymorphisms in the IL-18 gene promoter region. Reviewed International journal

Kouji Osawa, Takafumi Etoh, Noritaka Ariyoshi, Itsuko Ishii, Michteru Ohtani, Satoru Kariya, Katsuyoshi Uchino, Mitsukazu Kitada

The Journal of dermatology 34 ( 8 ) 531 - 6 2007.8

Language：English Publishing type：Research paper (scientific journal)

Kaposi's varicelliform eruption is the most important problem in treating patients with atopic dermatitis (AD) with tacrolimus ointment. It has been considered that Kaposi's varicelliform eruption occurs due to decreased levels of interleukin (IL)-18. The aim of this study was to examine the relationship between Kaposi's varicelliform eruption and genetic polymorphisms in the IL-18 gene. IL-18 gene promoter polymorphisms were analyzed in 21 AD patients treated with tacrolimus ointment and in 100 healthy volunteers. Six AD patients with Kaposi's varicelliform eruption during the treatment with tacrolimus ointment showed significantly higher frequency in G-to-C mutations at the IL-18 gene promoter region -137 compared with 15 AD patients without Kaposi's varicelliform eruption. The 15 AD patients without Kaposi's varicelliform eruption as well as 100 healthy volunteers did not have mutations of G-to-C at the IL-18 gene promoter region -137. These results suggest that the onset of Kaposi's varicelliform eruption following the treatment with tacrolimus ointment is associated with the mutation of G-to-C in the IL-18 gene promoter region -137, and that caution is required when using tacrolimus ointment for treating AD patients with this mutation.

院外処方内容に関するチェックシート機能の利用とデータベース構築

長谷川敦, 小林えり, 近藤夏未, 木村真春, 松島徹, 中村貴子, 中澤一純, 有吉範高, 北田光一

日本病院薬剤師会雑誌 43 ( 8 ) 1087 - 1089 2007.8

Language：Japanese Publisher：(一社)日本病院薬剤師会

千葉大学医学部附属病院(以下、当院)薬剤部では、当院における院内・院外処方せんについて相互作用の可能性のある医薬品が併用された場合や複数の診療科から重複して薬が処方された場合に、チェックシートが出力されるシステムを利用している。しかし、このチェックシートに基づいて疑義照会を行う際、過去に問い合わせた内容であるか否かが判断できないため、処方鑑査の質は向上したものの、業務の効率化という点では問題を残していた。そこで今回、院外処方に対するチェックシートに関する照会事項をデータベース化した。その結果、データベースを利用することで同一内容の疑義照会を回避でき、問い合わせを行うか否かの判断も容易となった。また、チェックシートを基に疑義照会を行うことで処方内容に変更が生じ、その後のチェックシートの出力枚数が減少し、さらには保険薬局からの問い合わせの減少にもつながり、業務の効率化に貢献できることがわかった。(著者抄録)

Identification and partial characterization of a novel CYP2C9 splicing variant encoding a protein lacking eight amino acid residues. Reviewed International journal

Noritaka Ariyoshi, Yoko Shimizu, Yukari Kobayashi, Hiroyoshi Nakamura, Hiromitsu Nakasa, Kazuyoshi Nakazawa, Itsuko Ishii, Mitsukazu Kitada

Drug metabolism and pharmacokinetics 22 ( 3 ) 187 - 94 2007.6

Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)

CYP2C9 is known as an enzyme responsible for the metabolism of various clinically important drugs. Recently, we cloned a cDNA corresponding to a CYP2C9 splicing variant (SV), which seemed to have an open reading frame of a protein with 482 amino acid residues. To investigate whether or not the SV can be translated as a functionally active protein, we expressed the CYP2C9SV in insect cells, and spectrophotometric and enzymatic properties were characterized. The CYP2C9SV protein showed a typical reduced CO-difference spectrum, indicating that the translated protein binds a heme moiety. However, CYP2C9SV did not metabolize tolbutamide or diclofenac at all, suggesting that the SV protein appeared to lack the ability to catalyze reactions mediated by CYP2C9. Although the CYP2C9SV mRNA was detected in all human liver samples examined in this study by real-time PCR, the level was generally low, ranging between 0.7 and 9.6% of the normal CYP2C9 mRNA. These results suggest that the CYP2C9SV protein is unlikely to contribute to CYP2C9 activities, although it appears to be expressed in most individuals.

Helices F-G are important for the substrate specificities of CYP3A7 Reviewed

Nao Torimoto, Itsuko Ishii, Ken-Ichi Toyama, Masayuki Hata, Kanako Tanaka, Hitoshi Shimomura, Hiroyoshi Nakamura, Noritaka Ariyoshi, Shigeru Ohmori, Mitsukazu Kitada

DRUG METABOLISM AND DISPOSITION 35 ( 3 ) 484 - 492 2007.3

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS

CYP3A7 is a member of the human CYP3A family and a major form of P450 expressed in human fetal livers. Although CYP3A7 shares nearly 90% base sequence with CYP3A4, CYP3A7 shows striking functional differences in the catalytic preference for several substrates, such as dehydroepiandrosterone ( DHEA) or dehydroepiandrosterone 3-sulfate (DHEA-3S). First, to clarify the reason for the differences between CYP3A7 and CYP3A4, a homology model of CYP3A7 was constructed using the CYP3A4 crystal structure. Because these two structures were similar, four kinds of chimeric enzymes were constructed to determine which sequences are important for exhibiting the characteristics of CYP3A7. The results of kinetic analysis of DHEA and DHEA-3S 16 alpha-hydroxylations by CYP3A7, CYP3A4, and CYP3A chimeras suggested that the amino acid residues from Leu210 to Glu279 were important to express the specificity for substrates as CYP3A7. This region was on the F and G helices of the modeled CYP3A7. Furthermore, to assess which amino acid in this sequence is important for the substrate specificity of CYP3A7, a one-point mutation of CYP3A7 to CYP3A4 was made by site-directed mutagenesis. The mutants of K224T and K244E had lost DHEA and DHEA-3S 16 alpha-hydroxylation activities. The mutants also greatly decreased the metabolism of testosterone, erythromycin, nevirapine, and triazolam relative to those activities of CYP3A7 wild-type enzyme. From these results, it is expected that CYP3A7 can recognize specific substrates using the lysines in F-G loops.

DOI： 10.1124/dmd.106.011304

Theoretical calculation of triazolam hydroxylation and endogenous steroid inhibition in the active site of CYP3A4. Reviewed International journal

Nao Torimoto, Itsuko Ishii, Masayuki Hata, Yukari Kobayashi, Hiroyoshi Nakamura, Noritaka Ariyoshi, Mitsukazu Kitada

Biochimica et biophysica acta 1774 ( 2 ) 223 - 32 2007.2

Language：English Publishing type：Research paper (scientific journal)

CYP3A4 has unusual kinetic characteristics because it has a large active site. CYP3A4 produced more 4-hydroxytriazolam than alpha-hydroxytriazolam at concentrations of more than 60 muM triazolam, and different steroids had different inhibitory effects on the system. To clarify these interesting observations, the interactions between substrate and substrate/steroid were investigated by theoretical calculations. When two triazolam molecules were docked into the active site, the distance between the O-atom and the 4-hydroxylated site was less than the distance to the alpha-hydroxylated site because of interaction between the two triazolam molecules. Estradiol inhibited both alpha- and 4-hydroxytriazolam formation by 50%. Dehydroepiandrosterone (DHEA) inhibited alpha-hydroxylation more than 4-hydroxytriazolam formation, whereas aldosterone had no effect. When one triazolam molecule and one steroid molecule were simultaneously docked, estradiol increased the distance between the O-atom and the two hydroxylated sites, DHEA only increased the distance between the O-atom and alpha-hydroxylated site, and aldosterone did not change the distances. The relevant angles of Fe-O-C in the hydroxylated site of triazolam also widened, together with increased distance. These findings indicate that formation of a substrate and substrate/effector complex in the active site may be a factor for determining the enzyme kinetic parameters of CYP3A4.

資料に基づく後発医薬品の評価プラバスタチンナトリウムおよびシンバスタチン製剤の比較

久保さやか, 中澤一純, 長谷川敦, 岡田賢二, 有吉範高, 北田光一

日本病院薬剤師会雑誌 42 ( 2 ) 219 - 223 2006.2

Language：Japanese Publisher：(一社)日本病院薬剤師会

資料に基づく後発医薬品の評価としてHMG-CoA還元酵素阻害剤のプラバスタチンナトリウムとシンバスタチン製剤を比較した.方法は,プラバスタチンナトリウム製剤22製品およびシンバスタチン製剤12製品の添付文書,インタビューフォームを用いて情報の評価を行なった.1)添付文書,インタビューフォーム記載事項はいずれも製剤間で異なっており,先発品は全ての調査項目について記載されていたが,後発品では吸収速度定数(Ka),消失速度定数(Ke),クリアランス(CL),分布容積(Vd)が記載されているのはプラバスタチンナトリウムで4割以下,シンバスタチンで5割以下であった.2)添加物では,両製剤とも9割以上の後発品で記載があったが,記載のないもの,製剤により添加物が違うものを認めた.3)安全性では,先発品ほど詳細な検討が行われていない後発品が過半数を占め,条件設定も製剤間で異なり,資料請求電話番号なしが7割を占めていた.4)単回投与での血中濃度推移は製剤間で大きなばらつきを認めた.以上,先発品も含め,情報提供のあり方に課題があると考えられた

CYP1A1 is a major enzyme responsible for the metabolism of granisetron in human liver microsomes Reviewed

H Nakamura, N Ariyoshi, K Okada, H Nakasa, K Nakazawa, M Kitada

CURRENT DRUG METABOLISM 6 ( 5 ) 469 - 480 2005.10

Language：English Publisher：BENTHAM SCIENCE PUBL LTD

Granisetron, a potent 5-HT3 receptor antagonist, has been reported to be mainly metabolized to 7-hydroxygranisetron and a lesser extent to 9'-desmethylgranisetron in humans. A previous study indicated that cytochrome P450 (CYP)3A4 is a major catalyst of 9'-demethylation, although the major CYP isoform(s) responsible for 7-hydroxylation are unknown. To clarify granisetron 7-hydroxylase, the in vitro metabolism of granisetron using expressed human CYPs and human liver microsomes was investigated. 7-Hydroxygranisetron was produced almost exclusively by CYP1A1, while, apparently, 9'-desmethylgranisetron was preferentially produced by CYP3A4. Marked inter-individual differences in the ratio of the formation of 7-hydroxygranisetron and 9'-desmethylgranisetron in human liver microsomes was observed. Granisetron 7-hydroxylase activity was strongly correlated with benzo[a]pyrene 3-hydroxylase activity (p < 0.0001), but not with testosterone 6 beta-hydroxylase activity in human liver microsomes. Furthermore, an anti-human CYP I A I antibody completely inhibited 7-hydroxylation in human liver microsomes, however, the reaction was not inhibited at all by an anti-CYP3A4 antibody. On the other hand, granisetron 9'-demethylase activity correlated significantly not only with testosterone 6 beta-hydroxylase activity (p < 0.0001) but also with benzo[a]pyrene 3-hydroxylase activity (p < 0.01). Consistent with this, both the anti-CYP1A1 and anti-human CYP3A4 antibodies inhibited the 9'-demethylase activity. These data indicate that CYP1A1 is a major enzyme responsible for the metabolism of granisetron via a main 7-hydroxylation pathway and an alternative 9'-demethylation route. This is the first report demonstrating the substantial contribution of CYP1A1 to the metabolism of a drug, although its role in the metabolism of environmental compounds is well established.

造血幹細胞移植施行患者におけるフルコナゾール血中濃度モニタリングの有用性に関する検討

鈴木貴詞, 岡田賢二, 中村裕義, 有吉範高, 中澤一純, 北田光一

TDM研究 22 ( 4 ) 322 - 327 2005.10

Language：Japanese Publisher：(一社)日本TDM学会

造血幹細胞移植(HSCT)施行患者に対し,フルコナゾール(FLCZ)は深在性真菌症予防の目的で高用量が長期間投与される.HSCT施行患者では免疫抑制剤など腎毒性の高い薬物が投与されるため,腎排泄型薬物であるFLCZの血中濃度の上昇を来し,副作用を生じる恐れがある.そこで,HSCT施行患者を対象として血漿中FLCZ濃度の測定を行い,FLCZ血中濃度モニタリングの有用性について検討を行った.今回対象とした7名の患者のいずれにおいても,腎機能低下に伴うと考えられる血漿中FLCZ濃度の上昇が認められたが,その上昇の程度は患者間で大きく異なっていた.現在FLCZの添付文書には,クレアチニンクリアランスが50ml/min以下の患者には投与量を半減するように記載されているが,本方法では,患者個々に対する至適投与量の設定は困難と推察された.したがって,腎機能の低下が頻発するHSCT施行患者におけるFLCZ血中濃度モニタリングの施行は適正使用の観点から望ましいと考えられた(著者抄録)

Inhibitory effects of nicardipine to cytochrome P450 (CYP) in human liver microsomes. Reviewed

Katsunori Nakamura, Noritaka Ariyoshi, Takafumi Iwatsubo, Yasuhisa Fukunaga, Saburou Higuchi, Kunio Itoh, Noriaki Shimada, Kazuo Nagashima, Tsuyoshi Yokoi, Koujirou Yamamoto, Ryuya Horiuchi, Tetsuya Kamataki

Biological & pharmaceutical bulletin 28 ( 5 ) 882 - 5 2005.5

Language：English Publishing type：Research paper (scientific journal)

To anticipate drug-drug interactions by nicardipine in vivo, cytochrome P450 (CYP) forms responsible for the metabolism of nicardipine and inhibition of CYP-dependent drug metabolism by nicardipine were investigated. Microsomes of human B-lymphoblastoid cells expressing each human CYP form were used for the metabolism of nicardipine. Inhibitory effects of nicardipine on drug metabolism were studied using human liver microsomes. CYP2C8, CYP2D6 and CYP3A4 were identified as major CYP forms for the metabolism of nicardipine in human liver microsomes. Nicardipine strongly inhibited two-pathways of triazolam hydroxylation both catalyzed by CYP3A4. Comparison of three Ca(2+) antagonists, nicardipine, nifedipine, and diltiazem revealed that only nicardipine showed such a strong inhibitory potency on the typical CYP2D6-catalyzed drug metabolism. Furthermore, nicardipine inhibited other reactions catalyzed by CYP1A, CYP2A6, CYP2C8, CYP2C9 and CYP2C19 with K(i) values ranging from 1.1 to 29.4 microM. In conclusion, nicardipine was a relatively potent inhibitor of human CYP2D6, CYP3A4 and CYP2C (especially for CYP2C8 and CYP2C19) in vitro, suggesting that drug-drug interactions between nicardipine and other drugs metabolized mainly by these CYP forms appear to occur in vivo.

血液透析患者におけるアルベカシンの投与設計

岡田賢二, 鈴木貴詞, 中村裕義, 石川耕, 有吉範高, 中澤一純, 齋藤康, 北田光一

TDM研究 22 ( 1 ) 27 - 33 2005.1

Language：Japanese Publisher：(一社)日本TDM学会

45歳男.2型糖尿病,慢性腎不全,右下腿潰瘍,MRSA骨髄炎であった.慢性腎不全による浮腫を軽減する目的で,週3回,1回4時間の血液透析(HD)を施行した.右下腿潰瘍部よりMRSAおよびP.aeruginosaを検出した.HDスケジュールを考慮したアルベカシン(ABK)の投与設計を試みた.HD施行3時間前にABKを投与することによって,ピーク濃度を約9〜10μg/mlで数時間持続させ,かつトラフ濃度を2μg/ml付近まで低下させることを可能にした.しかし,体温とCRPの低下は認めたが,MRSAの消失には至らなかった

心臓血管外科手術後患者に対するアンジオテンシン変換酵素阻害薬およびアンジオテンシンII受容体拮抗薬使用成績

公手妙, 中村貴子, 仲佐啓詳, 石井晃, 中澤一純, 有吉範高, 今牧瑞浦, 宮崎勝, 北田光一

日本病院薬剤師会雑誌 40 ( 11 ) 1425 - 1427 2004.11

Language：Japanese Publisher：(一社)日本病院薬剤師会

アンジオテンシン変換酵素阻害薬(ACEI)およびアンジオテンシンII受容体拮抗薬(ARB)は,高カリウム(K)血症をひき起こすことが知られ,K保持性利尿薬との併用や腎機能低下患者に対する使用では,高K血症を助長させる危険性があり注意が必要である.今回,心臓血管外科手術後の患者におけるACEIあるいはARBの使用と高K血症との関連性について検討した.ACEIおよびARB使用による明らかなK値上昇傾向は認めず,電解質変動への影響が少ない薬剤であることが示唆された.しかし,高K値を示した患者を個々に評価すると,これら薬剤による高K血症の発現が疑われる症例もあり,使用時には十分な注意が必要であると考えられた

Identification of deletion-junction site of CYP2A6*4B allele lacking entire coding region of CYP2A6 in Japanese Reviewed

N Ariyoshi, H Sekine, K Nakayama, K Saito, A Miyamoto, T Kamataki

PHARMACOGENETICS 14 ( 10 ) 701 - 705 2004.10

Effects of Continuous Ingestion of Green Tea or Grape Seed Extracts on the Pharmacokinetics of Midazolam Reviewed

NISHIKAWA Masataka, ARIYOSHI Noritaka, KOTANI Akira, ISHII Itsuko, NAKAMURA Hiroyoshi, NAKASA Hiromitsu, IDA Mayuri, NAKAMURA Hideo, KIMURA Nobuhito, KIMURA Maharu, HASEGAWA Atsushi, KUSU Fumiyo, OHMORI Shigeru, NAKAZAWA Kazuyoshi, KITADA Mitsukazu

Drug Metabolism and Pharmacokinetics 19 ( 4 ) 280 - 289 2004.8

Language：English Publisher：The Japanese Society for the Study of Xenobiotics

Limited systematic data on herb-drug interaction are available, despite many opportunities to concomitant use of herb with prescribed drugs. We investigated the effects of 15 herbal extracts in dietary supplements on CYP2C9, CYP2D6 and CYP3A4 activities in human liver microsomes. Strong inhibition of these CYP activities was found by the addition of green tea extracts (GTE) or grape seed extracts (GSE) in vitro. To examine the effects of these extracts on CYP3A activities in vivo, the pharmacokinetics of midazolam (MDZ) was analyzed in rats. Although single treatments with these extracts had negligible effects, 1 week of treatment with them resulted in a significant increase in the ke of intravenously administered MDZ, indicating the induction of CYP3A in the liver. In contrast, 1 week of treatment with GTE, but not GSE, caused a significant increase in the C_max and AUC_0-∞ of orally administered MDZ without change in the t_1/2, suggesting a reduction in CYP3A activity in the small intestines. These studies indicate that subchronic ingestion of GTE or GSE may alter the pharmacokinetics of MDZ, and the effects of GTE on CYP3A activity appear opposite between liver and small intestine, which could not be predicted from in vitro experiments.<br>

DOI： 10.2133/dmpk.19.280

Other Link： http://search.jamas.or.jp/link/ui/2006149345

The use of urine to clarify the genotype of a patient with toxic phenytoin concentrations who had undergone peripheral blood stem cell transplantation. Reviewed International journal

Noritaka Ariyoshi, Katsuhiro Sho-no, Miki Nishimura, Michihiro Ito, Hiroyoshi Nakamura, Takayoshi Asai, Yasushi Saitoh, Takashi Ishizaki, Mitsukazu Kitada

British journal of clinical pharmacology 58 ( 2 ) 225 - 6 2004.8

【健康食品・植物成分の薬物療法への影響】健康食品の薬物動態への影響

有吉範高

日本病院薬剤師会雑誌 40 ( 8 ) 955 - 957 2004.8

Direct interaction between substrates and endogenous steroids in the active site may change the activity of cytochrome P450 3A4 Reviewed

N Torimoto, Ishii, I, M Hata, H Nakamura, H Imada, N Ariyoshi, S Ohmori, T Igarashi, M Kitada

BIOCHEMISTRY 42 ( 51 ) 15068 - 15077 2003.12

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER CHEMICAL SOC

CYP3A4 exhibits unusual kinetic characteristics that result from the metabolism of multiple substrate including endogenous steroids and some drugs that coexist at the active site. To clarify the mechanism of the effect of endogenous steroids on the drug metabolism, the interaction between substrates, nevirapine (NVP) and carbamazepine (CBZ), and endogenous steroids was investigated by theoretical calculations. When the activities of NVP 2-hydroxylation and CBZ 10,11-epoxidation by expressed CYP3A4 were measured in the presence of steroids, NVP 2-hydroxylation was found to be remarkably increased by aldosterone and inhibited by estradiol. CBZ 10,11-epoxidation was increased by androstenedione. Three-dimensional computer modeling has shown that the active site of CYP3A4 is especially large, permitting access of two substrate molecules. The interactions between NVP and aldosterone and between CBZ and androstenedione were estimated by theoretical calculations assuming the substrate and steroids to be present in the active site at the same time. It was shown that NVP or CBZ would be stably fixed close to the oxygen atom at the sixth ligand of heme by interaction with steroids, suggesting that NVP and CBZ may be hydroxylated more easily due to the interaction with steroids. Estradiol was also expected to interact with NVP via a pi/pi interaction between a benzene ring, in which the NVP hydroxylation site is located, and a benzene ring of estradiol, suggested to inhibit the reaction. From these results, interactions between the substrate and endogenous steroids in the active site may change the activity of CYP3A4.

DOI： 10.1021/bi034409n

病院薬剤師と薬局薬剤師の連携実習型調剤技術懇話会の参加者の評価

中村安孝, 仲佐啓詳, 長谷川敦, 松島徹, 木村真春, 柴田みづほ, 三浦剛, 稲野祥宗, 大塚知子, 明石利恵子, 丹野恵子, 有吉範高, 中澤一純, 中村均, 大森栄, 北田光一

薬学雑誌 123 ( 6 ) 463 - 468 2003.6

CYP3A4 and CYP3A7-mediated carbamazepine 10,11-epoxidation are activated by differential endogenous steroids Reviewed

H Nakamura, N Torimoto, Ishii, I, N Ariyoshi, H Nakasa, S Ohmori, M Kitada

DRUG METABOLISM AND DISPOSITION 31 ( 4 ) 432 - 438 2003.4

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS

Recently, we reported that several endogenous steroids affect CYP3A4-mediated drug metabolism, using human adult liver microsomes as an enzyme source. Especially, carbamazepine (CBZ) 10,11-epoxidation is activated by androstenedione (AND). In the present studies, we investigated the effects of endogenous steroids on the activity of CBZ 10,11-epoxidation by expressed CYP3A4 and CYP3A7. When expressed CYP3A4 was used as an enzyme source, the addition of AND to the reaction mixture also caused a drastic increase in the activity of CBZ 10,11-epoxidase, and resulted in a change in the kinetics from sigmoid to Michaelis-Menten type. On the other hand, expressed CYP3A7-mediated CBZ 10,11-epoxidation was activated by sulfate conjugate steroids, such as pregnenolone 3-sulfate, 17alpha-hydroxypregnenolone 3-sulfate, and dehydroepiandrosterone 3-sulfate (DHEA-S), whereas the unconjugated form corresponding to these three steroids did not activate the reaction. Especially, DHEA-S was found to be a potent activator of CBZ 10,11-epoxidation by expressed CYP3A7. The kinetic character of CBZ 10,11-epoxidation by CYP3A7 is Michaelis-Menten type regardless of the presence of DHEA-S. The presence of DHEA-S caused a decrease in K-m and increase in V-max for CYP3A7-mediated CBZ 10,11-epoxidation, whereas DHEA-S 16alpha-hydroxylation was not affected by the coexistence of CBZ. In conclusion, CYP3A4 and CYP3A7-mediated CBZ 10,11-epoxidations are activated by different types of endogenous steroids. This is the first report regarding CYP3A7 cooperativity.

DOI： 10.1124/dmd.31.4.432

Association of CYP2A6 gene deletion with cigarette smoking status in Japanese adults Reviewed

Masahiko Ando, Nobuyuki Hamajima, Noritaka Ariyoshi, Tetsuya Kamataki, Keitaro Matsuo, Yoshiyuki Ohno

Journal of Epidemiology 13 ( 3 ) 176 - 181 2003

Language：English Publishing type：Research paper (scientific journal) Publisher：Japan Epidemiology Association

BACKGROUND: Genetic variation of CYP2A6 is shown to alter nicotine metabolism. This study was developed to investigate the genetic influence of the whole deletion-allele of CYP2A6 on active and passive smoking behavior. METHODS: Two hundred and forty Japanese adults, who visited Aichi Cancer Center as outpatients, were genotyped for the wild-type (CYP2A6*1A, CYP2A6*1B) and the whole deletion-type (CYP2A6*4C) polymorphism of CYP2A6. Information about active and passive smoking status was obtained by a self-administered questionnaire. Genetic influence of CYP2A6 polymorphism on smoking behavior was evaluated using the Mantel extension test. RESULTS: The frequency of the deletion allele was 18%. All 8 subjects carrying two deletion alleles had no smoking habit, and the homozygous deletion genotype showed a tendency to correlate with active smoking status after adjustment for sex and age (p=0.054). However, the proportion of never smokers among heterozygous subjects was almost the same as among subjects carrying no deletion allele (54% and 58%, respectively). Furthermore, CYP2A6 genotypes were correlated neither with the number of cigarettes smoked per day nor with the age at starting smoking (p=0.364 and 0.880, respectively). Among never smokers, CYP2A6 genotypes were not correlated with exposure to passive smoking at home or in the workplace (p=0.623 and 0.484, respectively). CONCLUSION: Despite the possible protection against active smoking behavior in subjects homozygous for the deletion allele, the CYP2A6 polymorphism has only a limited impact on public health because no protective effect was found in heterozygous subjects.

DOI： 10.2188/jea.13.176

New allelic arrangement CYP2D6*36x2 found in a Japanese poor metabolizer of debrisoquine Reviewed

M Chida, N Ariyoshi, T Yokoi, N Nemoto, M Inaba, M Kinoshita, T Kamataki

PHARMACOGENETICS 12 ( 8 ) 659 - 662 2002.11

Genetic polymorphism of CYP2A6 gene and tobacco-induced lung cancer risk in male smokers Reviewed

N Ariyoshi, M Miyamoto, Y Umetsu, H Kunitoh, H Dosaka-Akita, Y Sawamura, J Yokota, N Nemoto, K Sato, T Kamataki

CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION 11 ( 9 ) 890 - 894 2002.9

Authorship：Lead author Language：English Publishing type：Research paper (scientific journal) Publisher：AMER ASSOC CANCER RESEARCH

Cytochrome P450 2A6 (CYP2A6) is the principal enzyme involved in the metabolic activation of tobacco-specific nitrosamines to their ultimate carcinogenic forms and metabolism of nicotine. We investigated the effects of the CYP2A6*4, an entire CYP2A6 gene deletion-type polymorphism, on lung cancer risk and daily cigarette consumption in Japanese male smokers via a hospital-based case control study. The frequency of the CYP2A6*4 variant was compared in 370 lung cancer patients and 380 control smokers. A markedly reduced adjusted odds ratio for lung cancer risk, 0.23 [95 % confidence interval, 0.08-0.67], was seen in the group with homozygous deletion (*4/*4) when the odds ratio for a group with homozygous wild (*1A/*1A) was defined to be 1.00 by logistic regression. The subjects with lung cancer were additionally divided into three groups according to the histological classification of the cancer and examined for an association with the CYP2A6 polymorphism. The *4/ *4 genotype was not found in patients with squamous cell carcinoma (0 of 105) or small cell carcinoma (0 of 44), indicating that subjects with the *4/*4 genotype have low risk for lung cancers, particularly those caused by tobacco smoke. Furthermore, a significant reduction of daily cigarette consumption was observed in smokers with the *4/*4 genotype, suggesting a possibility that complete lack of CYP2A6 appeared to affect the smoking behavior. These data suggest that male smokers possessing the *1A/*1A genotype have higher risk for tobacco-induced lung cancers.

Characterization of a genotype previously designated as CYP2A6 D-type: CYP2A6*4B, another entire gene deletion allele of the CYP2A6 gene in Japanese Reviewed

N Ariyoshi, H Sekine, K Saito, T Kamataki

PHARMACOGENETICS 12 ( 6 ) 501 - 504 2002.8

Authorship：Lead author Language：English Publishing type：Research paper (scientific journal) Publisher：LIPPINCOTT WILLIAMS & WILKINS

CYP2A6 is known as an enzyme responsible for the metabolism of several clincally used drugs such as tegafur. Previously, we found two novel genotypes of the CYP2A6 gene, D-type and E-type, and the E-type was clarified to be homozygous for the CYP2A6* 4A allele. On the other hand, since the D-type was reported to lack regions from at least intron 5 to a part of exon 9 of the CYP2A6 gene, it caused a misunderstanding that the D-type would be a partial CYP2A6 gene-deleted allele. In this paper, we demonstrate that the D-type is a genotype heterozygous for the CYP2A6*4A and another novel entire CYP2A6 gene-deleted allele, CYP2A6*4B, by analyzing a Japanese family including parents genotyped as the CYP2A6*4A/4A and CYP2A6* 1A/*4B, respectively.

DOI： 10.1097/00008571-200208000-00012

A novel mutant allele of the CYP2A6 gene (CYP2A6*11) found in a cancer patient who showed poor metabolic phenotype towards tegafur Reviewed

S Daigo, Y Takahashi, M Fujieda, N Ariyoshi, H Yamazaki, W Koizumi, S Tanabe, K Saigenji, S Nagayama, K Ikeda, Y Nishioka, T Kamataki

PHARMACOGENETICS 12 ( 4 ) 299 - 306 2002.6

Language：English Publishing type：Research paper (scientific journal) Publisher：LIPPINCOTT WILLIAMS & WILKINS

In a clinical study, a newly developed anticancer drug, TS-1 capsule, which contained tegafur (FT) and 5-chloro-2,4-dihydroxypyridine, an inhibitor of dihydropyrimidine dehydrogenase, was orally administered to five gastric cancer patients (patients 1-5). The total area under the plasma FT concentration-time curve in patient 1 was fourfold higher than in other patients. Since cytochrome P450 2A6 (CYP2A6) has been reported to metabolize FT to yield 5-fluorouracil (5-FU), it was postulated that the poor metabolic phenotype of patient 1 was caused by mutations of the CYP2A6 gene. Thus, alleles for the CYP2A6 genes derived from patient 1 were completely sequenced. It was found that one allele was CYP2A6*4C, which was a whole deleted allele for the human CYP2A6 gene. The other allele was a novel mutant allele (CYP2A6*11) in which thymine at nucleotide 670 was changed to cytosine. The nucleotide change caused an amino acid change from serine at residue 224 to proline. To examine whether or not the amino acid change affected CYP2A6 activity, we expressed an intact or mutant CYP2A6 together with NADPH-P450 oxidoreductase in Escherichia coli, and compared the capacity of the wild and mutant enzymes to metabolize FT to 5-FU. The V-max value for FT metabolism by the mutant CYP2A6 was approximately one-half of the value of the intact CYP2A6, although the K-m values were nearly the same. From these results, we conclude that the poor metabolic phenotype of patient 1 was caused by the existence of the two mutant alleles, CYP2A6*4C and the new variant CYP2A6*11.

DOI： 10.1097/00008571-200206000-00005

Activation of microsomal epoxide hydrolase by interaction with cytochromes P450: kinetic analysis of the association and substrate-specific activation of epoxide hydrolase function Reviewed

K Taura, H Yamada, E Naito, N Ariyoshi, M Mori, K Oguri

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 402 ( 2 ) 275 - 280 2002.6

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

The kinetics of the association between cytochrome P450 (P450) and microsomal epoxide hydrolase (mEH) was studied by means of resonant mirror based on the principle of surface plasmon resonance. The dissociation equilibrium constants (KD) for the affinity of P450 enzymes for mEH were estimated by resonant mirror using an optical biosensor cell covalently bound to rat mEH. Comparable KD values were obtained for CYP1A1 and 2131, and these were greater by one order of magnitude than that for the CYP2C11. To clarify the influences of P450 enzymes on the catalytic activity of mEH, the hydrolyzing activity for styrene oxide and benzo(a)pyrene-7,8-oxide [B(a)P-oxide] was analyzed in the presence or absence of P450s. Styrene oxide hydrolysis was activated by all P450s including the CYP1A, 2B, 2C, and 3A subfamilies. In agreement with the association affinity determined by resonant mirror, CYP2C11 tends to have enhanced activity for styrene oxide hydrolysis. On the other hand, B(a)P-oxide hydrolysis was enhanced by only CYP2C11 while CYP1A1 and CYP2B1 had no effect. These results suggest that (1) many P450 enzymes associate nonspecifically with mEH, (2) the CYP2C11 plays a greater role in the association/activation of mEH and (3) the P450-mediated activation of mEH depends upon the substrate of mEH. (C) 2002 Elsevier Science (USA). All rights reserved.

DOI： 10.1016/S0003-9861(02)00079-6

Effects of endogenous steroids on CYP3A4-mediated drug metabolism by human liver microsomes Reviewed

H Nakamura, H Nakasa, Ishii, I, N Ariyoshi, T Igarashi, S Ohmori, M Kitada

DRUG METABOLISM AND DISPOSITION 30 ( 5 ) 534 - 540 2002.5

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS

In the present study, we investigated the effects of 14 endogenous steroids on the CYP3A4-mediated drug metabolism by human liver microsomes in vitro. Nevirapine (NVP) 2-, 12-hydroxylations, carbamazepine (CBZ) 10,11-epoxidation, triazolam (TZM) 1'-, 4-hydroxylations, erythromycin (EM) N-demethylation, and 2-sulphamoylacetylphenol (SMAP) formation from zonisamide (ZNS) were investigated. The activities of the NVP 2-, 12-hydroxylations, the CBZ 10,11-epoxidation, and the TZM 4-hydroxylation were activated by endogenous androgens, such as androstenedione (AND), testosterone, and dehydroepiandrosterone. However, these androgens inhibited EM N-demethylation, TZM 1'-hydroxylation, and SMAP formation. To understand the mechanisms of these effects of androgens on CYP3A4 activities, we performed a kinetic analysis of the metabolism of CBZ and ZNS in the presence or absence of AND using the modified two-site equation model. The addition of AND to the reaction mixture caused a drastic increase in the activity of CBZ 10,11-epoxidase, especially at a low substrate concentration, and resulted in a change in the kinetics from the sigmoid to Michaelis-Menten type. On the other hand, the metabolism of ZNS was strongly inhibited by AND, although no allosteric change was observed in this case. These data demonstrate that endogenous steroids, especially androgens, strongly affect CYP3A4-mediated drug metabolism in vitro. The postulated mechanisms of the interactions between AND and CBZ or ZNS are discussed.

DOI： 10.1124/dmd.30.5.534

CYP2D6.10 present in human liver microsomes shows low catalytic activity and thermal stability Reviewed

K Nakamura, N Ariyoshi, T Yokoi, S Ohgiya, M Chida, K Nagashima, K Inoue, T Kodama, N Shimada, T Kamataki

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 293 ( 3 ) 969 - 973 2002.5

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

Comparing bufuralol 1'-hydroxylase activity among liver microsomes prepared from individuals whose CYP2D6 genotypes had been determined, we found that the activity tended to decrease depending on the number of the CYP2D6*10 allele. Pre-incubation of liver microsomes from individuals homozygous for the CYP2D6*10 allele resulted in a decrease in the enzyme activity more rapidly than those from individuals homozygous for the CYP2D6*1, suggesting that not only the catalytic activity but also the thermal stability of the enzyme appeared to be affected by the genetic polymorphism. To confirm this hypothesis, the kinetic parameters of CYP2D6.1 and CYP2D6.10 were compared for bufuralol 1'-hydroxylation and dextromethorphan O-demethylation using microsomes prepared from yeast transformed with plasmids carrying CYP2D6 cDNAs (*1A and *10B). Kinetic studies of these CYP2D6 forms indicated clear differences in the metabolic activities between the wild (CYP2D6.1) and the mutant enzymes (CYP2D6.10). Bufuralol 1'-hydroxylase activity in microsomes of yeast expressing CYP2D6.10 was rapidly decreased by heat treatment. supporting the idea that the thermal stability of the enzyme was reduced by amino acid replacement from Pro (CYP2D6.1) to Ser (CYP2D6.10). These data strongly suggest that the thermal instability together with the reduced intrinsic clearance of CYP2D6.10 is one of the causes responsible for the known fact that Orientals show lower metabolic activities than Caucasians for drugs metabolized mainly by CYP2D6, because of a high frequency of CYP2D6*10 in Orientals. (C) 2002 Elsevier Science (USA). All rights reserved.

DOI： 10.1016/S0006-291X(02)00328-5

Variation in coumarin 7-hydroxylase activity associated with genetic polymorphism of cytochrome P450 2A6 and the body status of iron stores in adult Thai males and females. Reviewed International journal

Pailin Ujjin, Soisungwan Satarug, Yuvaree Vanavanitkun, Satoshi Daigo, Noritaka Ariyoshi, Hiroshi Yamazaki, Paul E B Reilly, Michael R Moore, Tetsuya Kamataki

Pharmacogenetics 12 ( 3 ) 241 - 9 2002.4

Language：English Publishing type：Research paper (scientific journal)

The relationships between catalytic activity of cytochrome P450 2A6 (CYP2A6), polymorphism of CYP2A6 gene, gender and levels of body iron stores were analysed in a sample group of 202 apparently healthy Thais, aged 19-47 years. Eleven individuals were found to have high activity of CYP2A6, judged by the relatively large amounts (11.2-14.6 mg) of 7-hydroyxcoumarin (7-OHC) excreted 3 h following administration of 15 mg of coumarin. Ten individuals, however, did not excrete any 7-OHC. Of these 10, four were found to have no CYP2A6 gene (whole gene deletion; CYP2A6*4 allele). The frequency of the CYP2A6 alleles; *1A, *1B and *4 in the whole sample group was 52, 40 and 8% while the frequency of the CYP2A6 gene types; *1A/*1A, *1A/*1B, *1B/*1B, *1A/*4, *1B/*4, *4/*4 was 29, 41, 16, 7, 5 and 2%. Subjects having CYP2A6*1A/*1B gene-type group were found to have higher rates of coumarin 7-hydroxylation compared with those of the CYP2A6*1B/*1B and CYP2A6*1A/*4 gene types. The inter-individual variability in CYP2A6 catalytic activity was therefore attributed in part to the CYP2A6 genetic polymorphism. Variation in CYP2A6 activity in this sample group was not associated with gender but, interestingly, it did show an inverse association with plasma ferritin; an indicator of body iron stores. Higher rates of coumarin 7-hydroxylation were found in individuals with low body iron stores (plasma ferritin < 20 microg/l) compared with subjects having normal body iron store status. Subjects (n = 16) with iron overload (plasma ferritin > 300 microg/l) also tended to have elevated rates of coumarin 7-hydroxylation. These results suggest an increased CYP2A6 expression in subjects who have excessive body iron stores. Further investigations into the underlying factors that may lead to increased expression of CYP2A6 in association with abnormal body iron stores are currently in progress in our laboratory.

CYP2A6 gene deletion reduces oral cancer risk in betel quid chewers in Sri Lanka Reviewed

Z Topcu, Chiba, I, M Fujieda, T Shibata, N Ariyoshi, H Yamazaki, F Sevgican, M Muthumala, H Kobayashi, T Kamataki

CARCINOGENESIS 23 ( 4 ) 595 - 598 2002.4

Language：English Publishing type：Research paper (scientific journal) Publisher：OXFORD UNIV PRESS

We investigated the relationship between inter-individual difference in CYP2A6 genotype and susceptibility to oral cancer among habitual betel quid chewers in a Sri Lanka population. A total of 286 subjects showing oral malignant or premalignant lesions and 135 control subjects with no lesions were analyzed. The frequency of homozygotes for CYP2A6*4C mutation, a gene deletion type of polymorphism, was significantly lower in the case subjects than the controls. The odds ratio (OR) of the group homozygous for the deletion was significantly lower and calculated to be 0.14 (95% CI; 0.03-0.72). In the allelic base analysis, there was also a significant decrease in the OR of the deletion allele. Our data suggest that deficient CYP2A6 activity due to genetic polymorphism reduces oral cancer risk in betel quid chewers.

Role of human cytochrome P450 (CYP) in the metabolic activation of nitrosamine derivatives: Application of genetically engineered Salmonella expressing human CYP

T Kamataki, K Fujita, K Nakayama, Y Yamazaki, M Miyamoto, N Ariyoshi

DRUG METABOLISM REVIEWS 34 ( 3 ) 667 - 676 2002

Language：English Publishing type：Research paper (scientific journal) Publisher：MARCEL DEKKER INC

The role of human cytochrome P450 (CYP) in the metabolic activation of tobacco-related N-nitrosamines was examined by Salmonella mutation test using a series of genetically engineered Salmonella typhimurium YG7108 strains each co-expressing a form of CYP (CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, and CYP3A5) together with human NADPH-cytochrome P450 reductase. Seven tobacco-related N-nitrosamines such as 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, N-nitrosodiethylamine, N-nitrosopyrrolidine, N-nitrosopiperidine, N-nitrosonornicotine, N-nitrosoanabasine, and N-nitrosoanatabine were used. The CYP2A6 was found to be responsible for the mutagenic activation of essentially all tobacco-related N-nitrosamines examined. On the basis of the evidence, genetic polymorphism of the CYP2A6 gene appeared to be one of the factors determining cancer susceptibility caused by smoking. Previously, we found the whole deletion of the CYP2A6 gene (CYP2A6*4C) as a type of genetic polymorphism in Japanese. We hypothesized that individuals possessing the gene homozygous for CYP2A6*4C were incapable of activating tobacco-related N-nitrosamines and showed lower susceptibility to lung cancer induced by tobacco smoke. Thus, the relationship between the CYP2A6*4C and the susceptibility to the lung cancer was evaluated. The frequency of the CYP2A6*4C was significantly lower in the lung cancer patients than healthy volunteers, suggesting that the subjects carrying the CYP2A6*4C alleles are resistant to carcinogenesis caused by N-nitrosamines because of the poor metabolic activation capacity.
Taking these results into account, CYP2A6 is an enzyme enhancing lung cancer risk.

DOI： 10.1081/DMR-120005668

The relationship between CYP2A6 gene deletion and susceptibility to oral cancer

Z Topcu, Chiba, I, M Fujieda, T Shibata, N Ariyoshi, H Yamazaki, F Sevgican, M Muthumala, H Kobayashi, T Kamataki

DRUG METABOLISM REVIEWS 34 154 - 154 2002

Role of the dioxin-like toxic compound coplanar polychlorinated biphenyl, 3,3 ',4,4 ',5-pentachlorobiphenyl in reducing hepatic alcohol dehydrogenase levels in rats in vivo

Y Ishii, H Kato, M Hatsumura, T Ishida, N Ariyoshi, H Yamada, K Oguri

JOURNAL OF HEALTH SCIENCE 47 ( 6 ) 575 - 578 2001.12

Language：English Publishing type：Research paper (scientific journal) Publisher：PHARMACEUTICAL SOC JAPAN

The ability of a dioxin-like toxic compound, coplanar polychlorinated biphenyl, 3,3',4,4',5-pentachlorobiphenyl (PCB126) to reduce the protein level of hepatic class 1 alcohol dehydrogenase (ADH), which plays an important role in the metabolism of ethanol, was studied. Male Wistar rats received PCB126 25 mg/kg i.p. At this dose the compound induces a wasting syndrome. PCB126 administration resulted in a significant suppression of the protein level of class 1 ADH, whereas the difference between free- and pair-fed controls was slight. These results suggest that dioxins also reduce class 1 ADH without involvement of decreased food consumption. These data offer new insights into the toxicity of dioxins via a marked decrease in the level of class 1 ADH.

DOI： 10.1248/jhs.47.575

Relationships between non-occupational cadmium exposure and expression of nine cytochrome P450 forms in human liver and kidney cortex samples Reviewed

Baker, JR, S Satarug, PEB Reilly, RJ Edwards, N Ariyoshi, T Kamataki, MR Moore, DJ Williams

BIOCHEMICAL PHARMACOLOGY 62 ( 6 ) 713 - 721 2001.9

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

This study was undertaken to assess associations between age, gender, cigarette smoke and non-workplace cadmium exposure, and liver pathology and inter-individual variation in cytochrome P450 (CYP) expression in human tissues. Autopsy specimens of twenty-eight Queensland residents whose ages ranged from 3 to 89 years were analyzed for the presence of nine CYP protein isoforms by immunoblotting. All subjects were Caucasians and their liver cadmium contents ranged from 0.11 to 3.95 kg/g wet weight, while their kidney cadmium contents were in the range of 2 to 63 mug/g wet weight. CYP1A2, CYP2A6, CYP2D6, CYP3A4, and CYP3A5 were detected in liver but not in kidney, and CYP1A1 and CYP1B1 were not found in liver or kidney. Lowered liver CYP2C8/19 protein contents were found to be associated with liver pathology. Importantly, we show elevated levels of CYP2C9 protein to be associated with cadmium accumulation in liver. No mechanism that explains this association is apparent, but there are two possibilities that require further study. One is that variation in CYP2C9 protein levels may be, in part, attributed to an individual's non-workplace exposure to cadmium, or an individual's CYP2C9 genotype may be a risk factor for cadmium accumulation. A positive correlation was found between liver CYP3A4 protein and subject age. Levels of liver CYPIA2 protein, but not other CYP forms, were increased in people more exposed to cigarette smoke, but there was no association between CYPIA2 protein and cadmium. CYP2A6 protein was found in all liver samples and CYP2A6 gene typing indicated the absence of CYP2A6 null allele (CYP2A6(D)) in this sample group, confirming very low prevalence of homozygous CYP2A6(D) in Caucasians. CYP2A6 gene types W/W, WIC, and CIC were not associated with variations in liver microsomal CYP2A6 protein. CYP2D6 protein was absent in all twenty-five kidney samples tested but was detectable in liver samples of all but two subjects, indicating the prevalence of the CYP2D6 null allele (CYP2D6(D)) in this sample group to be about 7%, typical of Caucasian populations. (C) 2001 Elsevier Science Inc. All rights reserved.

Comparison of the levels of enzymes involved in drug metabolism between transgenic or gene-knockout and the parental mice Reviewed

N Ariyoshi, S Imaoka, K Nakayama, Y Takahashi, K Fujita, Y Funae, T Kamataki

TOXICOLOGIC PATHOLOGY 29 161 - 172 2001.9

Language：English Publishing type：Research paper (scientific journal) Publisher：TAYLOR & FRANCIS INC

Drug-metabolizing enzymes are involved in the metabolic activation or detoxification of carcinogens. To evaluate animals developed as models for alternative carcinogenicity testing, we investigated whether or not a gene manipulation including the transgene of ras and the knocking out of a tumor suppressor gene such as p53 or XPA could alter the expression of representative drug-metabolizing enzymes directly or indirectly. Expression of several isoforms of cytochrome P450 (CYP) in the liver of rasH2, p53 (+/-), Tg. AC, and XPA (-/-) mice with or without treatment of prototype inducer, phenobarbital or 3-methylcholanthrene, was analyzed by Western immunoblotting in comparison with their parental strains of mice. In addition, the activities of 3 major phase II enzymes, UDP-glucronosyltransferase, sulfotransferase, and glutathione S-transferase, were compared between the gene-manipulated and the corresponding parental strains of mice. Results demonstrate that XPA gene knockout appeared to increase constitutive expression of CYP2B and CYP3A isoforms. Overexpression of human c-Ha-ras gene or p53 gene knockout appeared to increase constitutive UGT activity toward 4-nitrophenol. The content or activities of almost all other enzymes examined in the present study do not appear to be affected by the gene manipulation.

代謝研究は創薬にどう活かされるかレトロスペクティブな評価とプロスペクティブな活用 Cytochrome P450とmEH及びUGTとの蛋白質間相互作用

田浦健一郎, 山田英之, 内藤絵里, 竹田修三, 石井祐次, 有吉範高, 森正明, 小栗一太

薬物動態 16 ( Suppl. ) S94 - S95 2001.9

Language：Japanese Publisher：(一社)日本薬物動態学会

ラット肝を用いて異種薬物代謝酵素間における蛋白-蛋白相互作用の有無を検討した.その結果,小胞体膜上のcytochrome P450はmEH(microsomal epoxide hydrolase)やUGT(UDP-glucuronosyltransferase)等の異種薬物代謝酵素と相互作用することで多段階の薬物代謝反応を効率的に実行している可能性が示唆された

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Relationships between non-occupational cadmium exposure and expression of nine cytochrome P450 forms

Baker, JR, S Satarug, PEB Reilly, RJ Edwards, N Ariyoshi, T Kamataki, DJ Williams, MR Moore

TOXICOLOGY 164 ( 1-3 ) 61 - 62 2001.7

CYP遺伝子多型解析の病棟業務への応用

斉藤嘉津彦, 清水瓊子, 岡崎正子, 伊林至洋, 端和夫, 前野康次郎, 石井清二, 土橋和文, 島本和明, 戸田貴大, 黒澤菜穂子, 大和田栄治, 加藤芳伸, 大山徹, 梅津有理, 千田道洋, 有吉範高, 鎌滝哲也, 板谷幸一

医療薬学 27 ( 3 ) 228 - 234 2001.6

Language：Japanese Publisher：(一社)日本医療薬学会

脳神経外科で初めて多型解析を利用した例として,脳動静脈奇形の術後に痙攣発作予防の目的で投与されたフェニトインのTDMを行った32歳女の例を提示し,経過及び薬物代謝酵素遺伝子解析後の服薬指導内容を述べた.また薬物代謝酵素遺伝子解析を希望した患者に対する情報提供として使用している「解析結果報告書」及び「解析結果の説明書」を紹介した

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コプラナーPCBによる中間代謝への影響ラット肝サイトソルのトリオースリン酸含量の低下について(Effects of a Highly Toxic Coplanar Polychlorinated Biphenyl, 3,3',4,4',5-Pentachlorobiphenyl on Intermediary Metabolism: Reduced Triose Phosphate Content in Rat Liver Cytosol)

石井祐次, 加藤晴敏, 初村恵, 石田卓巳, 有吉範高, 山田英之, 小栗一太

福岡医学雑誌 92 ( 5 ) 190 - 200 2001.5

Language：English Publisher：福岡医学会

トリオースリン酸代謝酵素群(グリセルアルデヒド-3-リン酸デヒドロゲナーゼ,トリオースリン酸イソメラーゼ,グリセロキナーゼ,トランスアルドラーゼ,及びトランスケトラーゼ)に及ぼす3,3',4,4',5-pentachlorobiphenyl(PenCB)の影響を調べた.又,PenCBによるアルコール脱水素酵素(ADH)の発現抑制が,どの程度トリオースリン酸代謝に影響を及ぼしているかを検討した.PenCB処理により,これら酵素群の活性はいずれも50%程度まで有意に低下していた.ピラゾールで阻害される活性をADH由来とした場合,ADHは約30%の寄与があることが分かり,特にこの部分への抑制が顕著であった.摂餌対照群とPenCB処理群では肝臓中のトリオースリン酸の含量が,自由摂餌群に比べて著しく低かった.PenCB処理によってトリオースリン酸代謝の回転が落ち,栄養を補給してもエネルギーとして利用されにくいシステムが出来ている可能性がある

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A novel single nucleotide polymorphism altering stability and activity of CYP2A6

N Ariyoshi, Y Sawamura, T Kamataki

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 281 ( 3 ) 810 - 814 2001.3

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC

CYP2A6 is known as a major cytochrome P450 (CYP) responsible for the oxidation of nicotine and coumarin in humans. In this study, we explored genetic polymorphisms, which reduce CYP2A6 activity in Japanese. Two novel mutations in exon 9 of the CYP2A6 gene were found. A single nucleotide polymorphism of T1412C and G1454T resulted in Ile471Thr and Arg485Leu substitution, respectively. The frequency of the former variant allele was considerably high (15.7%), while the latter variant appeared to be a rare polymorphism. Heterologous expression of CYP2A6 using a cDNA possessing C instead of T-base at codon 471 in Escherichia coli caused remarkable reduction of the stability of holoenzyme at 37 degreesC. Furthermore, this variant enzyme almost lacked nicotine C-oxidase activity, although coumarin 7-hydroxylase activity was still observed. These data suggest that individuals homozygous for the T1412C variant allele or heterozygous for this and a defect allele such as the CYP2A6*4 may be poor metabolizer of nicotine, but not coumarin. (C) 2001 Academic Press.

DOI： 10.1006/bbrc.2001.4422

A single nucleotide polymorphism of CYP2B6 found in Japanese enhances catalytic activity by autoactivation

N Ariyoshi, M Miyazaki, K Toide, Y Sawamura, T Kamataki

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 281 ( 5 ) 1256 - 1260 2001.3

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC

A single nucleotide polymorphism (SNP) resulting in a substitution from Gin to His was found in exon 4 of the CYP2B6 gene in Japanese. The frequency of the variant allele was found to be 19.9%. The mutant- and the wild-type enzymes were expressed in Escherichia coli, and the effects of the single amino acid substitution on the catalytic activity were examined by investigating the kinetic profiles of 7-ethoxycoumarin O-deethylase activity. The wild-type enzyme showed typical Michaelis-Menten kinetics, while the mutant-type enzyme represented the sigmoidal kinetics with a higher V-max value compared to that of the wild-type enzyme. Eadie-Hofstee plots further revealed an existence of allosteric effects for the reaction catalyzed by the variant. This is the first evidence demonstrating that only one amino acid substitution, Gln172His, caused by natural SNP enhances the catalytic activity of CYP by obtaining the character of homotropic cooperativity. stool Academic Press.

DOI： 10.1006/bbrc.2001.4524

Genetic polymorphisms of cytochrome P450 2A6 in a case-control study on lung cancer in a French population

MA Loriot, S Rebuissou, M Oscarson, S Cenee, M Miyamoto, N Ariyoshi, T Kamataki, D Hemon, P Beaune, Stucker, I

PHARMACOGENETICS 11 ( 1 ) 39 - 44 2001.2

Language：English Publishing type：Research paper (scientific journal) Publisher：LIPPINCOTT WILLIAMS & WILKINS

Cytochrome P450 2A6 (CYP2A6) is involved in the C-oxidation of nicotine and in the metabolic activation of tobacco nitrosamines. Recent data have suggested that CYP2A6 genetic polymorphisms might play a role in tobacco dependence and consumption as well as in lung cancer risk, However, the previously published studies were based on a genotyping method that overestimated the frequencies of deficient alleles, leading to misclassification for the CYP2A6 genotype, In this study, we genotyped DNA from 244 lung cancer patients and from 250 control subjects for CYP2A6 (wild-type allele CYP2A6*1, and two deficient alleles: CYP2A6*2, and CYP2A6*4, the latter corresponding to a deletion of the gene) using a more specific procedure. In this Caucasian population, we found neither a relation between genetically impaired nicotine metabolism and cigarette consumption, nor any modification of lung cancer risk related to the presence of defective CYP2A6 alleles (odds ratio = 1.1, 95% confidence interval = 0.7-1.9). Pharmacogenetics 11:39-44 (C) 2001 Lippincott Williams & Wilkins.

DOI： 10.1097/00008571-200102000-00005

Accumulation of the 1-methyl-4-phenylpyridinium ion in suncus (Suncus murinus) brain: Implication for flavin-containing monooxygenase activity in brain microvessels

T Mushiroda, N Ariyoshi, T Yokoi, E Takahara, O Nagata, H Kato, T Kamataki

CHEMICAL RESEARCH IN TOXICOLOGY 14 ( 2 ) 228 - 232 2001.2

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER CHEMICAL SOC

The metabolism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was examined in an effort to evaluate the role of flavin-containing monooxygenase (FMO) expressed in the brain of suncus (Suncus murinus) and rats. MPTP was metabolized to generate both 1-methyl-4-phenylpyridinium ion (MPP+) and MPTP N-oxide by brain homogenates from rats. Although the level of MPP+-producing activity was similar in suncus and rats, a remarkable difference was found between the animal species in MPTP N-oxygenase activity, which was not detectable in brain homogenates from suncus, The concentrations of MPP+ in suncus brain after a single ip administration of MPTP were markedly higher than that in rats, probably because of the lack of FMO activity in the suncus brain. The MPTP N-oxygenase activity of microvessel homogenates of rat brain was 21-fold greater than that of whole brain homogenates. These results suggest that FMO(s) plays a significant role in the detoxification of MPTP in cerebral endothelial cells.

DOI： 10.1021/tx0001225

INTERACTION OF CYTOCHROME P450 AND MICROSOMAL EPOXIDE HYDROLASE AND UDP-GLUCURONOSYLTRANSFERASE

TAURA Ken-ichiro, YAMADA Hideyuki, NAITO Eri, TAKEDA Shuso, ISHII Yuji, ARIYOSHI Noritaka, MORI Masa-aki, OGURI Kazuta

Drug Metabolism and Pharmacokinetics 16 94 - 95 2001

Language：English Publisher：The Japanese Society for the Study of Xenobiotics

Interaction between cytochrome P450 (P450) and other sorts of drug metabolizing enzymes was studied by affinity chromatography with CYP1A1-conjugated column. Solubilized rat liver microsomes were charged onto the column and the eluate was analyzed by immunoblotting. The result showed that the CYP1A1 column effectively trapped microsomal epoxide hydrolase (mEH) and UDP-glucuronosyltransferase (UGT) as well as NADPH-P450 reductase. Non-drug metabolizing enzymes such as calnexin and protein disulfide isomerase did not show any affinity to the column. Neither BSA- nor glycine-conjugated column could trap microsomal protein. These results strongly suggest the specific interaction between P450 and mEH/UGT. When estimated using stereo-selective hydrolysis for styrene oxide as the index, the function of rat mEH was enhanced by co-incubation with CYP1A1, 2B1 and 2C11. The same picture was also seen for the interaction between dog mEH and CYP2B11/3A12. These results suggest that P450 interacts with mEH and UGT to facilitate a series of multistep metabolic conversion.

DOI： 10.2133/dmpk.16.supplement_94

Structural characterization of a new variant of the CYP2A6 gene (CYP2A6*1B) apparently diagnosed as heterozygotes of CYP2A6*1A and CYP2A6*4C

N Ariyoshi, Y Takahashi, M Miyamoto, Y Umetsu, S Daigo, T Tateishi, S Kobayashi, Y Mizorogi, MA Loriot, Stucker, I, P Beaune, M Kinoshita, T Kamataki

PHARMACOGENETICS 10 ( 8 ) 687 - 693 2000.11

Language：English Publishing type：Research paper (scientific journal) Publisher：LIPPINCOTT WILLIAMS & WILKINS

During the course of investigating the frequency of a CYP2A6 whole deletion-type polymorphism (CYP2A6*4C) in Japanese, an unexpectedly large population of heterozygotes for CYP2A6*4C and the wild-type (CYP2A6*1A) was found. Cloning of a cDNA encoding CYP2A6 from the liver of individuals judged as heterozygotes for CYP2A6*4C and the CYP2A6*1A was carried out to identify the causal allele(s) responsible for a possible overestimation. A clone isolated from the liver cDNA library possessed 58 bp sequences in the 3'-untranslated region, which was replaced with the corresponding region of the CYP2A7 gene. The same gene conversion existed in the genomic DNA, indicating that the replacement was not a cloning artifact. Based on the gene structure of the allele (CYP2A6*1B), this variant was thought to be one of the causal alleles responsible for overestimation of heterozygotes for CYP2A6*4C and CYP2A6*1A. To investigate this further, we developed a genotyping method which could distinguish the CYP2A6*1A, CYP2A6*1B and CYP2A6*4C alleles from each other. The results clearly showed that CYP2A6*1B was the sole allele responsible for the overestimation. We conclude that the new genotyping method allows determination of six genotypes of the CYP2A6 gene, simultaneously and precisely, in both Oriental and Caucasian populations. Pharmacogenetics 10:687-693 (C) 2000 Lippincott Williams & Wilkins.

DOI： 10.1097/00008571-200011000-00003

A high-level expression of CYP2A in the lung of the suncus (Suncus murinus) and its role in the activation of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone

T Mushiroda, N Ariyoshi, K Kimura, E Takahara, O Nagata, H Kato, T Kamataki

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 269 ( 2 ) 393 - 396 2000.3

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC

Northern blot analysis of mRNA prepared from the lung of Suncus murinus (suncus), which was classified as an ancestor of primates, revealed that the expression level of cytochrome P450 2A (CYP2A) mRNA was about 100-fold higher than in the lung from rats and mice. To confirm that the pulmonary CYP2A of the suncus had a catalytic activity, the metabolism of a specific substrate for CYP2A6, (+)-cis-3,5-dimethyl-2-(3-pyridyl) thiazolidin-4-one hydrochloride (SM-12502), was determined. The intrinsic clearance for SM-12502 S-oxidation by the suncus lung microsomes was calculated to be 99-fold higher than that by rat liver microsomes. The mutagen-producing activity of a 9000g supernatant fraction prepared from suncus lung was examined using 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) as a promutagen. The results showed that the suncus lung possessed 82-fold higher mutagen-producing activity than the rat lung, indicating that NNK was efficiently activated by the CYP2A isoform expressed in the suncus lung and that the suncus was a sensitive animal species to the genotoxicity of NNK contained in tobacco smoke. (C) 2000 Academic Press.

DOI： 10.1006/bbrc.2000.2312

日本人と欧米人における CYP2A6 遺伝子多型の解析

宮本昌美, 梅津有理, 醍醐聡, BEAUNE Philippe, LORIOT Marie-Anne, STUCKER Isabelle, 木下盛敏, 名倉弘哲, 立石智則, 小林真一, 溝呂木能浩, 有吉範高, 鎌滝哲也

臨床薬理 31 ( 2 ) 301 - 302 2000.2

A highly toxic coplanar polychlorinated biphenyl compound suppresses Delta 5 and Delta 6 desaturase activities which play key roles in arachidonic acid synthesis in rat liver Reviewed

K Matsusue, Y Ishii, N Ariyoshi, K Oguri

CHEMICAL RESEARCH IN TOXICOLOGY 12 ( 12 ) 1158 - 1165 1999.12

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER CHEMICAL SOC

The effect of 3,3',4,4',5-pentacklorobiphenyl (PenCB) on the synthesis of unsaturated fatty acids was studied in male Wistar rats. The arachidonic acid (20:4) content in the total lipids of liver homogenates was significantly reduced on day 5 of PenCB administration, while those of linoleic acid (18:2) and bishomo-gamma-linolenic acid (20:3) were increased. These changes in the total lipids of liver homogenates were observed following doses of PenCB ranging from 0.5 to 25 mg/kg of body weight. The same changes in these fatty acids were seen with four subtypes of microsomal glycerophospholipids in the liver. The marked reduction in the molar ratio of 20:4 to 18:2 in the lipids suggests alteration of the activity of the enzymes responsible for the synthesis of unsaturated fatty acids. The activity of Delta 5 and Delta 6 desaturases (arachidonic acid synthetase) in the liver microsomes was 17 and 13% of that of pair-fed control animals, whereas the activity of 1-acylglycerophosphorylcholine or 1-acylglycerophosphate acyltransferase, which transfers 20:4 or 18:2 to phospholipids, was not affected by the treatment. Thus, the reduction in the level of 20:4 that was observed can be explained by a reduction in desaturase activity. These results are evidence that the coplanar PenCB has a significant effect on the reduced synthesis of physiologically essential long-chain unsaturated fatty acids.

Analysis of the CYP2D6 gene in relation to dextromethorphan O-demethylation capacity in a Japanese population (vol 65, pg 570, 1999)

T Tateishi, M Chida, N Ariyoshi, Y Mizorogi, T Kamataki, S Kobayashi

CLINICAL PHARMACOLOGY & THERAPEUTICS 66 ( 6 ) 581 - 581 1999.12

CYP2A6 gene deletion reduces susceptibility to lung cancer

M Miyamoto, Y Umetsu, H Dosaka-Akita, Y Sawamura, J Yokota, H Kunitoh, N Nemoto, K Sato, N Ariyoshi, T Kamataki

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 261 ( 3 ) 658 - 660 1999.8

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC

CYP2A6 is an enzyme with a high ability to activate a nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), to its potent and ultimate carcinogen. In the present study, we investigated the relationship between genetic polymorphism of CYP2A6 and lung cancer risk in a case-control study of Japanese subjects. Genotyping of the CYP2A6 gene in both healthy volunteers and lung cancer patients was conducted. The frequency with which the subjects carried homozygotes of the CYP2A6 gene deletion-type mutation (deletion), which causes lack of the enzyme activity, was lower in the lung cancer patients than in the healthy control subjects. The odds ratio (OR) of the group homozygous for the deletion was significantly lower and calculated to be 0.25 (95% CI; 0.08-0.83) when the OR for the population with homozygotes of the CYP2A6 wild-type gene was defined as 1.00, In the allelic-base analysis, there was also a significant decrease in the OR for the deletion allele, These data suggest that deficient CYP2A6 activity due to genetic polymorphism reduces lung cancer risk. (C) 1999 Academic Press.

DOI： 10.1006/bbrc.1999.1089

THE STUDY ON CYP2A6 GENETIC POLYMORPHISM

ARIYOSHI Noritaka, NUNOYA Ken-ichi, TAKAHASHI Yuki, UMETSU Yuri, MIYAMOTO Masami, DAIGO Satoshi, YOKOI Tsuyoshi, KWON Jun-Tack, KADLUBAR Fred F., SHINHA Rashimi, BEAUNE Philippe, KIMURA Kanzo, KAMATAKI Tetsuya

Drug Metabolism and Pharmacokinetics 14 100 - 101 1999

Language：English Publisher：The Japanese Society for the Study of Xenobiotics

CYP2A6 is known as an enzyme responsible for the metabolism of coumarin. As its toxicological significance, aflatoxin B₁ and NNK are metabolically activated by this CYP to mutagens. In our recent studies, we found that a newly developed drug, SM-12502, was a specific substrate of CYP2A6, and 3 out of 28 Japanese were poor metabolizers (PM) of this drug. Genomic analysis of CYP2A6 gene in the PM indicated that an existence of a new CYP2A6 gene variant which lacks the entire region of open reading frame for the enzyme. The frequency of the individual who possess the mutation homozygously was estimated as 3-4% in Japanese. During the course of investigation of the frequency, we found an another CYP2A6 variant whose 60 by region in the 3'-untranslated region was substituted by the corresponding sequences of CYP2A7 gene. In this study, we investigated the frequency of these polymorphic alleles as well as reported v1 (CYP2A6*2) variant among different race.

DOI： 10.2133/dmpk.14.supplement_100

Metabolism of 2,3 ',4 ',5-tetrachlorobiphenyl by cytochrome P450 from rats, guinea pigs and hamsters Reviewed

N Koga, N Kikuichi, T Kanamaru, H Kuroki, K Matsusue, C Ishida, N Ariyoshi, K Oguri, H Yoshimura

CHEMOSPHERE 37 ( 9-12 ) 1895 - 1904 1998.10

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

The metabolism of 2,3',4',5-tetrachlorobiphenyl (TCB) was compared using liver microsomes and six isoforms of cytochrome P450 purified from rats, guinea pigs and hamsters. In microsomal study, the following species differences were observed: 1) Untreated guinea pigs and hamsters but not rats can metabolize this TCB to 3-hydroxy- or 4-hydroxy-2,3',4',5-TCB, 2) Guinea pig microsomes showed only 3-hydroxylating activity, whereas hamster microsomes showed higher activity of 4-hydroxylation than that of 3-hydroxylation. In common with three species, the 3-hydroxylation was accelerated by phenobarbital. The 4-hydroxylation in rats and hamsters was increased by pretreatment with 3-methylcholanthrene and 3,3',4,4',5-pentachlorobiphenyl. The hydroxylation activities of liver microsomes from the three species could be explained by an involvement of different isoforms of cytochrome P450. In addition, it is apparent that hamster CYP1A2 as well as hamster CYP2A8 is involved in the 4-hydroxylation of 2,3',4',5-TCB although it has no activity for 2,2',5,5'-TCB. (C) 1998 Elsevier Science Ltd. All rights reserved.

Highly toxic coplanar PCB126 reduces liver peroxisomal enzyme activities in rats

N Ariyoshi, M Iwasaki, H Kato, S Tsusaki, M Hamamura, T Ichiki, K Oguri

ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY 5 ( 3 ) 219 - 225 1998.5

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

The effect of the highly toxic coplanar PCB congener, 3,4,5,3',4'-pentachlorobiphenyl (PCB126) on hepatic peroxisomes was studied in rats. The aim of this study was to investigate whether a toxic dose of the dioxin-like coplanar PCB modifies enzyme activities in peroxisomes where plays an important role in lipid metabolism. Treatment with PCB126, at a single i.p. administration of 25 mg/kg which evokes clear suppression of body weight gain, resulted in marked reduction (to about 40-50%) of catalase activity and peroxisomal fatty acyl-CoA beta-oxidizing system. Immunoblotting showed that expression of catalase was greatly reduced by the treatment in parallel with the activity. Light microscopy revealed a drastic reduction in granules possessing peroxidase activity, while electron microscopy demonstrated that no apparent morphological changes had taken place. Thus the reduction in catalase activity caused by PCB126 could be attributable to suppression of protein expression. The marked reduction of these peroxisomal enzyme activities might be related to hyperlipidemia caused by dioxin-related compounds in rats and humans. (C) 1998 Elsevier Science B.V. All rights reserved.

DOI： 10.1016/S1382-6689(98)00007-6

Metabolism of antihistaminics, promethazine and CYP2D6 genetic polymorphism.

Jpn. J. Clin. Pharmacol. Ther. 29 ( 1 ) 237 - 238 1998.3

Characterization of the rat Star gene that encodes the predominant 3.5-kilobase pair mRNA - ACTH stimulation of adrenal steroids in vivo precedes elevation of Star mRNA and protein Reviewed

N Ariyoshi, YC Kim, Artemenko, I, KK Bhattacharyya, CR Jefcoate

JOURNAL OF BIOLOGICAL CHEMISTRY 273 ( 13 ) 7610 - 7619 1998.3

Authorship：Lead author Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

The steroidogenic acute regulatory protein (STAR) participates in steroidogenesis through the mitochondrial transfer of cholesterol to cytochrome P450scc. The rat adrenal Star gene is transcribed as a 3.5-kilobase pair (kb) and 1.6 kb mRNA with the larger mRNA predominating (similar to 85% of total) in. vivo, Hypophysectomy (HPX) produced a 3-5-fold decrease in Star mRNA along with a loss of adrenal steroids, whereas P450scc mRNA decreased by less than 2-fold, Adrenocorticotropic hormone (ACTH) treatment of HPX rats maximally stimulated steroidogenesis rates within 5 min with over 10-fold elevation of steady state blood levels occurring within 10 min. For intact rats there was a 5-10-fold larger increase, paralleling previously observed elevations of cholesterol-cytochrome P450scc association and metabolism in subsequently isolated adrenal mitochondria, ACTH did not increase either total STAR protein or a group of modified forms until at least 30 min after completion of acute stimulation, indicating that elevated translation of STAR protein cannot alone mediate this acute stimulation. Parallel slow changes in STAR protein and corticosterone formation after ACTH treatment are consistent with participation of STAR forms as co-regulators of these hormonal responses, ACTH stimulation of HPX rats increased Star mRNA by 2.5-fold within 20 min and by 4.5-fold after 1 h, thus preceding the rise in the STAR protein, A 3.5-kb Star cDNA clone isolated from a rat adrenal cDNA library exhibited a 0.9-kb open reading frame and a 2.5-kb 3'-untranslated region (3'-UTR). The open reading frame sequence differed at only 12 amino acids from that of the mouse Star. The rat Star gene seven exons with exon 7 encoding the entire 2.5 kb of 3'-UTR of the 3,5-kb mRNA. The 3'-UTR sequence suggests that 1.6- and 3,5-kb mRNA are formed by an alternative usage of different polyadenylation signals, Multiple UUAUUUA(U/A)(U/A) motifs also suggest additional regulation through this extended 3'-UTR, Although elevation of STAR protein by ACTH does not cause the acute increase in adrenal cholesterol metabolism, changes in the turnover or distribution of an active STAR subfraction cannot be excluded.

DOI： 10.1074/jbc.273.13.7610

Species-specific alteration of hepatic glucose 6-phosphate dehydrogenase activity with coplanar polychlorinated biphenyl: Evidence for an Ah-receptor-linked mechanism

M Hori, H Kondo, N Ariyoshi, H Yamada, K Oguri

CHEMOSPHERE 35 ( 5 ) 951 - 958 1997.9

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

We examined the in vivo effect of a highly toxic coplanar polychlorinated biphenyl (PCB) on the hepatic activity of glucose 6-phosphate dehydrogenase (G6PDH) in aryl hydrocarbon (Ah)-responsive (C57/BL) and -less-responsive (DBA) strains of mice. The activity in the C57BL strain was moderately increased by 3,3',4,4',5-pentachlorobiphenyl (PCB 126) in a dose dependent manner. However, this was not observed in DBA mice although greater doses were injected. 2,2',5,5'-Tetrachlorobiphenyl (PCB 52) with a non-planar structure did not increase G6PDH activity, The increase in G6PDH activity with PCB 126 was also seen in rats, but not in guinea pigs. The activity in the latter species was decreased rather than increased. These results suggest that the induction of hepatic G6PDH by coplanar PCB is mediated by a mechanism involving the Ah receptor, and the response was highly species-specific. (C) 1997 Elsevier Science Ltd.

DOI： 10.1016/S0045-6535(97)00181-1

Changes in the hepatic glutathione peroxidase redox system produced by coplanar polychlorinated biphenyls in Ah-responsive and -less-responsive strains of mice: mechanism and implications for toxicity

M Hori, H Kondo, N Ariyoshi, H Yamada, A Hiratsuka, T Watabe, K Oguri

ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY 3 ( 4 ) 267 - 275 1997.9

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

The alteration in hepatic glutathione peroxidase (GPx) produced by polychlorinated biphenyls (PCBs) was studied in vivo in aryl hydrocarbon (Ah)-responsive C57BL and -less-responsive DBA strains of mice. 3,3',4,4',5-Pentachlorobiphenyl (PCB 126), one of the high-affinity ligands for the Ah receptor, significantly reduced Se-dependent GPx activity in C57BL mice, but not in DBA mice. A reduction in activity in C57BL mice was also observed following treatment with a high dose of 3,3',4,4'-tetrachlorobiphenyl with lesser affinity for the Ah receptor than PCB 126, but not by 2,2',5,5'-tetrachlorobiphenyl, a low-affinity ligand. To assess the effects on GPx in the liver, the content of reduced glutathione (GSH), an obligate co-factor for GPx, and the activity of two enzymes, gamma-glutamyl transpeptidase (gamma-GTP) and glutathione reductase (GR), which play a role in supplying GSH were determined after PCB treatment. The results showed that although the hepatic activity of gamma-GTP and GR was affected differently by PCB 126, the content of GSH was slightly increased rather than reduced in both strains of mice. The activity of non-Se-dependent GPx, which is due to the catalysis by some isozymes of glutathione S-transferase (GST), was significantly increased only in C57BL mice by PCB 126 treatment. Immunoblot analysis demonstrated that the induction of the class theta GST, which is a potent reducer of peroxides (Hiratsuka et al., 1995. Biochem. Biophys. Res. Commun. 212, 743) reflects the enhancement of the above activity. These results suggest that (i) the PCB-induced reduction in Se-dependent GPx activity is mediated by a mechanism involving the Ah receptor; and (ii) a concomitant increase in the class theta GST partially rescues the Ah-responsive mice from coplanar PCB-induced oxidative stress. (C) 1997 Elsevier Science B.V.

DOI： 10.1016/S1382-6689(97)00025-2

Co-planar PCBの肝glutathione peroxidase-redox systemに対する影響ラット及びモルモットでの検討

堀美穂, 有吉範高, 山田英之

福岡医学雑誌 88 ( 5 ) 144 - 148 1997.5

Language：Japanese Publisher：福岡医学会

活性酸素消去系の損傷とそれに基づく酸化的ストレスの役割について検討した.この目的の為,本研究ではラットとモルモットのcoplanar PCBに対する感受性の違いに注目した.即ち活性酸素消去系の障害が動物種を越えた共通の毒性発現機構なら,感受性動物のモルモットの方が感受性の低いラットよりも障害の程度が顕著なはずであるとの作業仮説を立てて検討を行ったが,上記の推定を支持する結果は得られなかった.即ち,PCB 126処理ラットでは主要な活性酸素消去系の一つであるGPx活性が有意に低下したが,感受性の高いモルモットではむしろ活性増加した.肝GSH含量並びにこの供給に働くGR及びγ-GTP活性の変動もラットでのみ障害の発生が観察された

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A highly toxic PCB produces unusual changes in the fatty acid composition of rat liver

K Matsusue, Y Ishii, N Ariyoshi, K Oguri

TOXICOLOGY LETTERS 91 ( 2 ) 99 - 104 1997.4

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCI IRELAND LTD

The changes in lipid metabolism produced by a coplanar PCB were studied in rats. Male Wistar rats were given a single intraperitoneal injection of 3,3',4,4',5-pentachlorobiphenyl at a dose of 25 mg/kg. After 5 days of administration, total hepatic lipids were treated with 1 M KOH in methanol at 75 degrees C and the liberated fatty acids were analyzed by HPLC after conversion to fluorescent derivatives. In comparison with free-fed and pair-fed control groups, the proportion of arachidonic acid in the PenCB-treated rats was reduced by about 50%, while oleic and linoleic acids increased significantly. We also examined the individual glycerophospholipids, separated by TLC,to see if they were affected by alteration in the fatty acid composition of the whole liver. In all glycerophospholipids, the proportion of arachidonic acid was reduced significantly to the same degree while linoleic acid increased. Changes in the activity of desaturase isozymes have been postulated to explain this unusual lipid metabolism following administration of a toxic PCB and this may contribute to its toxicity. (C) 1997 Elsevier Science Ireland Ltd.

DOI： 10.1016/S0378-4274(97)03881-2

Effect of highly toxic 3,3',4,4',5-pentachlorobiphenyl on the synthesis of unsaturated fatty acids Reviewed

K Matsusue, N Ariyoshi, M Inoue, Y Ishii, K Oguri

JAPANESE JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH 43 ( 1 ) P19 - P19 1997.2

Significant suppression of aldolase B, carbonic anhydrase III and alcohol dehydrogenase in liver cytosol of rats treated with a highly toxic coplanar PCB Reviewed

H Kato, Y Ishii, M Hatsumura, T Ishida, Nakayama, I, N Ariyoshi, K Oguri

JAPANESE JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH 43 ( 1 ) P20 - P20 1997.2

Significant suppression of rat liver aldolase B by a toxic coplanar polychlorinated biphenyl, 3,3',4,4',5-pentachlorobiphenyl

Y Ishii, H Kato, M Hatsumura, T Ishida, N Ariyoshi, K Oguri

TOXICOLOGY 116 ( 1-3 ) 193 - 199 1997.1

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCI IRELAND LTD

A toxic coplanar polychlorinated biphenyl, 3,3',4,4',5-pentachlorobiphenyl (PenCB), significantly suppresses the expression of liver aldolase B in rats, Hepatic aldolase activity in PenCB-treated rats was significantly reduced to about 50% of that in free- and pair-fed control groups. The reduced aldolase activity following PenCB-treatment was due to the marked suppression of the expression of aldolase B shown by immunoblot analysis after SDS-polyacrylamide gel electrophoresis and two-dimensional gel electrophoresis. The suppression of rat liver aldolase B could be a key biochemical lesion caused by PenCB. Copyright (C) 1997 Elsevier Science Ireland Ltd.

DOI： 10.1016/S0300-483X(96)03543-3

Cytochrome P450scc in rat adrenal cells mediated by regulation of the steroidogenic acute regulatory protein

YC Kim, N Ariyoshi, Artemenko, I, ME Elliott, KK Bhattacharyya, CR Jefcoate

STEROIDS 62 ( 1 ) 10 - 20 1997.1

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE INC

Cholesterol conversion to pregnenolone by cytochrome P450scc in steroidogenic cells, including those of the adrenal cortex is determined by hormonal control of cholesterol availability. Intramitochondrial cholesterol movement to P450scc, which retains hormonal activation in isolated mitochondria, is apparently dependent an peripheral benzodiazepine receptor and the recently cloned steroidogenic acute regulatory (SrARJ protein. In rat adrenal cells, StAR is formed as a 37-kDa precursor that is transferred to the mitochondrial inner membrane following phosphorylation by hormonally activated protein kinase A, and processed to multiple forms, some of which turn over very rapidly. In bovine cells, StAR undergoes three modifications for ming a set of eight proteins seen in both glomerulosa and fasciculata cells. In the former, cyclic AMP and angiotensin ii each decrease two forms and elevate six forms. Significantly, the major change seen after activation may not involve phosphorylation of StAR. Cholesterol transfer across mitochondrial membranes is also activated in isolated mitochondria by GTP and low concentrations of Ca2+, apparently prior to activation by StAR. Depletion of StAR by cyclohexmide inhibits cholesterol transfer but is overcome by uptake of Ca2+ into the matrix. This activation of cellular- cholesterol transport is sustained in adrenal cells pereabilized by Streptolysin 0. In rat adrenal cells cAMP elevates 3.5- and 1.6-kb mRNA, hybridized by a 1.O-kb StAR cDNA. A 3.5-kb rat adrenal cDNA that encodes all except the 5' end of the longest StAR mRNA has been characterized. The corresponding gene sequence is distributed across seven exons. The shorter mRNA may arise from polyadeylation signals early in exon 7. However, the 3.5-kb mRNA comprises 80-90% of untreated rat adrenal StAR mRNA and may therefore provide the prime source for in vivo translation of StAR protein. (C) 1997 by Elsevier Science, Inc.

DOI： 10.1016/S0039-128X(96)00153-5

Effect of co-planar polychlorinated biphenyl on the hepatic glutathione peroxidase redox system in rats and guinea pigs

M. Hori, N. Ariyoshi, H. Yamada, K. Oguri

Fukuoka Acta Medica 88 ( 5 ) 12 - 16 1997

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Kyushu University School of Medicine

The effect of 3,3',4,4',5-pentachlorobiphenyl (PCB 126) on hepatic glutathione peroxidase (GPx) redox system was studied in vivo in rats and guinea pigs. PCB 126 treatment caused significant reduction of Se-dependent and -non-dependent GPx activity in rats. In agreement with this, the content of glutathione (GSH) and the activities of GSH reductase (GR) and γ-glutamyl transpeptidase (γ-GTP) were also decreased in this species. On the contrary, guinea pig liver Se-non-dependent GPx activity was significantly enhanced by PCB 126 treatment, while no effect on Se-dependent activity was observed. Neither the content of GSH nor the enzyme activities responsible for GSH supply in guinea pig liver was affected by PCB 126. These result suggested that the damage on GPx redox system is, at least, one of mechanisms by which co-planar PCB induces the toxicity in rats. However, in guinea pigs, this is not the case, and different mechanism from the damage on active oxygen quenching system is likely to be involved.

Metabolism of 2,4,5,2',4',5'-hexachlorobiphenyl (PCB153) in guinea pig Reviewed

N. Ariyoshi, N. Koga, H. Yoshimura, K. Oguri

Xenobiotica 27 ( 9 ) 973 - 983 1997

Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)

1. The in vitro and vivo metabolism of 2,4,5,2',4',5'-hexachlorobiphenyl (PCB153) in guinea pig has been studied. 2. Seven metabolites were detected in the faeces of PCB153-treated animals and three were identical to those produced by dog liver microsomes. The detection of a metabolite where a chlorine atom was shifted from the 2- to 3-position strongly suggested the involvement of 2,3-arene oxide intermediate, and evidence for the concomitant formation of a 3,4-arene oxide intermediate was provided by identifying other two minor metabolites which were dechlorinated at the 4-position. 3. In vitro studies using liver microsomes from guinea pigs revealed that the 2,3-arene oxide and 5-hydroxylation pathways are the predominant metabolic routes compared with the 3,4-arene oxide pathway. Although the guinea pig is an another species that can metabolize PCB153 mainly to the 2,3-arene oxide intermediate, the rate of formation was only about one-tenth of the dog. 4. These results indicate that the ability to form this unusual 2,3-arene oxide intermediate may not be responsible for high excretion rate of this congener. Our data also suggest that the cytochrome P450-catalysed metabolism of PCB153 in the guinea pig and dog are similar, whereas for post-cytochrome P450 metabolism, the guinea pig resembles the rabbit.

DOI： 10.1080/004982597240136

Production of antibody against cytosolic 54kDa protein in rat liver evidence of the significant induction by a highly toxic coplanar polychlorinated biphenyl

T. Ishida, Y. Ishii, K. Tasaki, N. Ariyoshi, K. Oguri

Fukuoka Acta Medica 88 ( 5 ) 3 - 11 1997

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Kyushu University School of Medicine

We have reported that a 54kDa protein in rat liver cytosol is highly inducible by treatment with 3,3',4,4',5-pentachlorobiphenyl (PenCB) or 3- methylcholanthrene (MC) using SDS-polyacrylamide gel electrophoresis. Internal amino acid sequences of this protein in the rat liver were highly homologous to those of selenium binding protein (SeBP) or acetaminophen binding protein (APBP) in mouse liver cytosol. In this paper, the purification and characterization of this protein were demonstrated MC was given at a dose of 20 mg/kg for 3 consecutive days. The liver cytosolic 54kDa protein was purified twice from the MC-treated male Wistar rats by Rotofore Cell(TM) procedure to apparent single on SDS-polyacrylamide gel electrophoresis, and the rabbit antiserum against this protein was obtained. Male Wistar rats were given PenCB in corn oil at a single dose of 25 mg/kg i.p. The liver cytosol was prepared on the 5th day after the treatment and subjected to immunoblot analysis. The 54kDa protein was markedly induced in the liver cytosol of PenCB-treated rats. Immunoblot analysis after two- dimensional gel electrophoresis suggested that there could be isoforms of 54kDa protein. The induction of the 54kDa protein with PenCB was assumed to be mediated through Ah-receptor. The physiological role of the 54kDa protein was discussed together with SeBP and APBP, the role of which has not yet been elucidated.

Significant induction of a 54-kDa protein in rat liver with homologous alignment to mouse selenium binding protein by a coplanar polychlorinated biphenyl, 3,4,5,3',4'-pentachlorobiphenyl and 3-methylcholanthrene

Y Ishii, M Hatsumura, T Ishida, N Ariyoshi, K Oguri

TOXICOLOGY LETTERS 87 ( 1 ) 1 - 9 1996.9

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCI IRELAND LTD

A 54-kDa protein in rat liver cytosol was significantly induced by treatment with 3,4,5,3',4'-pentachlorobiphenyl (25 mg/kg, single i.p.) and 3-methylcholanthrene (20 mg/kg, once a day for 3 days, i.p.). The protein exhibited pi of 6.8 on two-dimensional gel electrophoresis. The amino acid sequences of peptide fragments from the protein digested in situ were highly similar to a selenium binding protein in mice and to the isoform acetaminophen binding protein in mice. The present result clearly demonstrates that a coplanar polychlorinated biphenyl and 3-methylcholanthrene are responsible for induction of selenium binding protein homologues. The physiological role of the mouse proteins, however, is not yet elucidated.

DOI： 10.1016/0378-4274(96)03668-5

Hamster liver cytochrome P450 (CYP2A8) as a 4-hydroxylase for 2,5,2',5'-tetrachlorabiphenyl Reviewed

N Koga, N Kikuichi, T Kanamaru, N Ariyoshi, K Oguri, H Yoshimura

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 225 ( 2 ) 685 - 688 1996.8

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS

Metabolism of 2,5,2',5'-tetrachlorobiphenyl (TCB) was studied using liver microsomes of hamsters and two hamster P450 isoforms, CYP1A2 and 2A8. CYP2A8 catalyzed selectively 4-hydroxylation of 2,5,2',5-TCB at a rate of 21.7 pmol/min/nmol P450. In contrast, CYP1A2 showed no activity for hydroxylation of 2,5,2',5'-TCB. Immunological study revealed that rabbit antiserum against CYP2A8 almost completely inhibited the microsomal 4-hydroxylation but that against CYP1A2 did not. It was also shown that the induction pattern of CYP2A8 protein by P450 inducer was similar to that of the 4-hydroxylase activity in hamster liver microsomes. These results suggest that CYP2A8 plays a major role in the 4-hydroxylation of 2,5,2',5'-TCB in hamster liver. (C) 1996 Academic Press, Inc.

A coplanar PCB induces a selenium binding protein as a major cytosolic protein in rat liver

Y Ishii, M Hatsumura, T Ishida, N Ariyoshi, K Oguri

CHEMOSPHERE 32 ( 3 ) 509 - 515 1996.2

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

We obtained evidence that a toxic coplanar polychlorinated biphenyl (PCB) induces a counterpart of murine 56kDa selenium binding protein in rat liver cytosol. A 54kDa protein in the liver cytosol was significantly induced by 3,3',4,4',5-pentachlorobiphenyl and proved to be a major cytosolic protein in the rat liver. The protein exhibited pi of 6.8 on two-dimensional gel electrophoresis. The amino acid sequence of peptide fragments from the protein digested in situ, was highly similar to a 56kDa selenium binding protein and similar to an acetaminophen binding protein in mice.

DOI： 10.1016/0045-6535(95)00316-9

Involvement of cytochrome b(5) in the metabolism of tetrachlorobiphenyls catalyzed by CYP2B1 and CYP1A1

K Matsusue, N Ariyoshi, K Oguri, N Koga, H Yoshimura

CHEMOSPHERE 32 ( 3 ) 517 - 523 1996.2

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

The role of cytochrome b(5) in the cytochrome P450 (CYP)-dependent hydroxylation of tetrachlorobiphenyl (TCB) isomers was examined using a reconstituted mixed function oxygenase (MFO) system containing purified CYP2B1 or 1A1, and rat liver microsomes. Hydroxylations of 2,2',5,5'- and 3,3',4,4'-TCBs were catalyzed mainly by CYP2B1 and 1A1, respectively, in the reconstituted MFO system and those of 2,3',4',5- and 2,3',4,4'-TCBs were mediated by both cytochrome P450 systems. The activity toward 2,2',5,5'- and 2,3',4',5-TCB was significantly increased 6.5- and 5.5-fold, respectively, by addition of cytochrome b, in the reconstituted MFO system containing of CY2B1. Either hydroxylation activity toward 2,3',4,4'-TCB with the CYP2B1 system was very low or decreased by addition of cytochrome b(5). These results suggest that the involvement of cytochrome b(5) to the hydroxylation of TCBs is dependent on the TCB congener being metabolized, and the cytochrome P450 isoform involved in its metabolism.

DOI： 10.1016/0045-6535(95)00318-5

METABOLISM OF HIGHLY PERSISTENT PCB CONGENER, 2,4,5,2',4',5'-HEXACHLOROBIPHENYL, BY HUMAN CYP2B6

N ARIYOSHI, K OGURI, N KOGA, H YOSHIMURA, Y FUNAE

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 212 ( 2 ) 455 - 460 1995.7

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS

Metabolism of 2,4,5,2',4',5'-hexachlorobiphenyl was studied with cDNA-expressed human P450 2B isoform, CYP2B6. 3-Hydroxy-2,4,5,2',4',5'-hexachlorobiphenyl was identified as a major metabolite, and the formation activity was compared with that of dog CYP2B11 and guinea pig P450(GP-1). The activity of 3-hydroxylation was comparable with that of P450(GP-1), but one-tenth of CYP2B11. These results indicate that P450 2B in humans as well as other animal species can metabolize 2,4,5,2',4',5'-hexachlorobiphenyl, and the reason why this PCB congener remained most abundantly in human bodies is discussed. (C) 1995 Academic Press, Inc.

DOI： 10.1006/bbrc.1995.1991

EFFECT OF HIGHLY TOXIC COPLANAR PCB ON PEROXISOMAL ENZYME-ACTIVITY - THE SPECIES-DIFFERENCE BETWEEN RATS AND GUINEA-PIGS Reviewed

M IWASAKI, H KATO, N ARIYOSHI, K OGURI

JAPANESE JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH 41 ( 1 ) P32 - P32 1995.2

Effect of a coplanar PCB on lipid metabolism ; The remarkable difference between rats and Guinea pigs(共著)

Fukuoka Acta Media 86 ( 5 ) 135 1995

Alteration of perixosomal enzyme activities in the liver of guinea pigs caused by coplanar PCB

M. Iwasaki, H. Kato, N. Ariyoshi, K. Oguri

Fukuoka Acta Medica 86 ( 5 ) 12 - 20 1995

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Kyushu University School of Medicine

The hyperlipidemia is a well-known typical symptom in Yusho patients and experimental animals treated with PCBs. We have found a significant induction of CYP4A1, which catalyzes ω-hydroxylation of fatty acids, in guinea pigs by the treatment with a coplanar PCB, 3,4,5,3',4'-pentachlorobiphenyl (PenCB), though the P450 is reduced in the treated rats. Peroxisome has β-oxidation enzymes distinct from mitochondrial enzymes, and also play an important role in lipid metabolism. Peroxisome proliferators have been shown to regulate the expression of CYP4A1 and peroxisomal enzymes by the same mechanism in the rat. In the present study, we examined the effect of PenCB treatment on peroxisomal enzymes in the liver of guinea pigs. As a result, the enzyme activities of hepatic peroxisome, e.g. fatty acid oxidizing system, catalase and urate oxidase, had a rising tendency by the treatment with PenCB in the animal. The results suggest that the regulation of peroxisomal enzymes and CYP4A1 is also associated in guinea pigs, and PenCB provides a similar effect of peroxisomal proliferators to the animal. The possible toxicity through the peroxisomal alteration was discussed.

Studies on PCB toxicity involving 2C subfamily cytochrome P450

N. Ariyoshi, S. Ito, A. Okudaira, M. Mise, K. Matsusue, H. Yamada, K. Oguri

Fukuoka Acta Medica 86 ( 5 ) 21 - 30 1995

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Kyushu University School of Medicine

It has been demonstrated that PCB metabolism is mainly catalyzed by 1A and 2B subfamily cytochrome P450s, CYP 1A1/2 and CYP 2B1/2. These studies were conducted mostly with hepatic enzymes in rodents. The 1A and 2B subfamily P450s are constitutively expressed little, but markedly induced by xenobiotics such as 3-methylcholanthrene and phenobarbital in rodents. On the other hand, the recent studies showed that cytochrome P450s in human liver are remarkably different from isoform of rodents in constitution and enzyme activities. In the present study, we first tried to metabolize some PCBs with 2C subfamily cytochrome P450 (CYP2C) purified from dog liver microsomes. The data suggested that CYP2C may not be involved in PCB metabolism. Since CYP2C is the same most abundant enzyme as 3A subfamily P450 in human liver and plays a major role for metabolism of many drugs used clinically, and may also play an important role for metabolism of some steroid hormones, we further studied the inhibition of CYP2C-catalyzed steroid metabolism by typical PCB congeners. CYP2C-mediated steroid metabolism is greatly inhibited by 2,4,5,2',4',5'-hexachlorobiphenyl, but not by 3,4,5,3',4'-pentachlorobiphenyl. On the contrary, 3,4,5,3',4'-pentachlorobiphenyl markedly suppressed CYP2C expression in the dog liver. These results suggest that residual PCBs may affect the current situation of steroid hormones in Yusho patients, and may cause PCB-drug interactions.

PURIFICATION AND CHARACTERIZATION OF DOG LIVER MICROSOMAL EPOXIDE HYDROLASE Reviewed

N ARIYOSHI, M TANAKA, Y ISHII, K OGURI

JOURNAL OF BIOCHEMISTRY 115 ( 5 ) 985 - 990 1994.5

Authorship：Lead author Language：English Publishing type：Research paper (scientific journal) Publisher：JAPANESE BIOCHEMICAL SOC

An epoxide hydrolase (mEH) in liver microsomes was purified to apparent homogeneity from a dog treated with phenobarbital. The purified enzyme had a minimum molecular weight of 47,000 as determined by SDS-PAGE. The dog mEH activity was characterized by use of a substrate, 7-glycidoxycoumarin (GOC), and some effecters of this enzyme. In vitro activators, metyrapone, and isoquinoline, stimulated the microsomal activity, but the former had no such effect on the purified enzyme in case of this substrate. All mEH inhibitors, 1,1,1-trichloropropene 2,3-oxide (TCPO), cyclohexene oxide, and 2-bromo-4'-nitroacetophenone (BrNAP), suppressed hydrolase activity. The NH2-terminal amino acid sequence of the purified enzyme was highly homologous (90%) to the sequences deduced from a cDNA clone of rat enzyme. Antiserum to the purified enzyme raised in rabbits cross-reacted with rat and guinea pig epoxide hydrolases. No gender-difference in this enzyme in liver microsomes was observed in dogs.

EFFECT OF COPLANAR PCB ON GLUCONEOGENESIS AND LIPID-METABOLISM - COMPARISON BETWEEN RATS AND GUINEA-PIGS Reviewed

M HATSUMURA, T ISHIDA, Y ISHII, N ARIYOSHI, Y KOGA, K OGURI, H YOSHIMURA

JAPANESE JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH 40 ( 1 ) P27 - P27 1994.2

INDUCTION OF BILIRUBIN UDP-GLUCURONOSYLTRANSFERASE AND CYP4A1 P450 BY COPLANAR PCBS - DIFFERENT RESPONSIVENESS OF GUINEA-PIGS AND RATS

Y KOGA, M TSUDA, N ARIYOSHI, Y ISHII, H YAMADA, K OGURI, Y FUNAE, H YOSHIMURA

CHEMOSPHERE 28 ( 3 ) 639 - 645 1994.2

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

Induction of hepatic microsomal bilirubin UDP-glucuronyltransferase and CYP4A1 P450 by 3,4,3',4'-tetrachlorobiphenyl (TCB) and 3,4,5,3',4'-pentachlorobiphenyl (PenCB) was studied in guinea pigs and rats. Both enzyme activities in guinea pigs were increased 4-fold or more over the control by PenCB treatment, while by treatment with a less toxic congener, TCB, these were increased to a lesser extent. On the contrary, the rat enzymes were significantly decreased by treatment with these co-planar polychlorinated biphenyls. These results suggest that the alterations of both bilirubin UDP-glucuronyltransferase and CYP4A P450 are the characteristic indices for the toxicities of polychlorinated hydrocarbons.

DOI： 10.1016/0045-6535(94)90305-0

MODIFICATION OF THE GLUCONEOGENESIS IS NOT INVOLVED IN THE COPLANAR PCB TOXICITY IN HIGHLY SENSITIVE GUINEA-PIGS Reviewed

K OGURI, M HATSUMURA, Y ISHII, Y KOGA, N ARIYOSHI, H YOSHIMURA

CHEMOSPHERE 27 ( 11 ) 2295 - 2303 1993.12

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

Effect of a co-planar PCB, 3,4,5,3',4'-pentachlorobiphenyl (PenCB) on gluconeogenesis in guinea pigs was compared with that in rats. The key enzyme of gluconeogenesis, phosphoenolpyruvate carboxykinase (PEPCK), in liver cytosol of rats was reduced by PenCB treatment to about 50% of control level. Hypoglycemia was also seen in the rat. On the contrary, the liver cytosol PEPCK activity in guinea pigs was slightly increased by PenCB treatment. Plasma levels of alanine and tryptophan were increased in the rat treated with PenCB, but the level of glycogenic amino acid, alanine, was not raised significantly-in the guinea pig. Thus the modification of the gluconeogenesis was not involved in the PenCB toxicity in highly sensitive guinea pigs.

DOI： 10.1016/0045-6535(93)90140-Z

IDENTIFICATION OF IN-VITRO METABOLITES OF 2,4,6,2',4',6'-HEXACHLOROBIPHENYL FROM PHENOBARBITAL-TREATED DOG LIVER-MICROSOMES Reviewed

N ARIYOSHI, H YOSHIMURA, K OGURI

BIOLOGICAL & PHARMACEUTICAL BULLETIN 16 ( 9 ) 852 - 857 1993.9

Authorship：Lead author Language：English Publishing type：Research paper (scientific journal) Publisher：PHARMACEUTICAL SOC JAPAN

We studied in vitro metabolites of 2,4,6,2',4',6'-hexachlorobiphenyl (HCB, IUPAC PCB No. 155) produced by liver microsomes of a phenobarbital (PB)-treated beagle dog. The major metabolites were 3-hydroxy-2,4,6,2',4',6'-HCB (M-1), 4-hydroxy-2,6,2',4',6'-pentachlorobiphenyl (PenCB, M-2) and 3,4-dihydroxy-2,6,2',4',6'-PenCB (M-3). Furthermore, 4-hydroxy-2,3,6,2',4',6'-HCB (M4), which could be formed via the 3,4-epoxidation and the subsequent NIH-shift of the chlorine from the 4 to the 3 position, was also detected. We found that M-3 is a common secondary metabolite of the two major monohydroxy metabolites, M-1 and M-2. These results indicate that the dog seems to metabolize and eliminate this congener not only by a mechanism involving direct insertion of a hydroxyl group but also via an arene oxide intermediate.

DOI： 10.1248/bpb.16.852

INDUCTION OF BILIRUBIN UDP-GLUCURONOSYLTRANSFERASE AND P-450-4A SUBFAMILY IN GUINEA-PIGS BY TREATMENT WITH COPLANAR PCB Reviewed

Y KOGA, M TSUDA, N ARIYOSHI, Y ISHII, H YAMADA, K OGURI, H YOSHIMURA, Y FUNAE

JAPANESE JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH 39 ( 1 ) P23 - P23 1993.2

Induction of Bilirubin UDP-Glucuronyltransferase and P-450 4A Subfamily in Guinea Pigs Reviewed

Yoshiko Koga, Yoshihiko Funaec, Hidetoshi Yoshimura, Noritaka Ariyoshi, Yuji Ishii, Hideyuki Yamada, Kazuta Oguri, Minoru Tsuda

Eisei kagaku 39 ( 1 ) 1993

Inducing Ability of Co-planar PCBs toward Bilirubin UDP-glucuronyltransferase of Liver Microsomes : The Remarkable Difference between Guinea Pigs and Rats. Reviewed

Fukuoka Acta Medica 84 ( 5 ) 175 1993

METABOLISM OF 2,4,5,2',4',5'-HEXACHLOROBIPHENYL WITH LIVER-MICROSOMES OF PHENOBARBITAL-TREATED DOG - THE POSSIBLE FORMATION OF PCB 2,3-ARENE OXIDE INTERMEDIATE Reviewed

N ARIYOSHI, N KOGA, K OGURI, H YOSHIMURA

XENOBIOTICA 22 ( 11 ) 1275 - 1290 1992.11

Authorship：Lead author Language：English Publishing type：Research paper (scientific journal) Publisher：TAYLOR & FRANCIS LTD

1. Metabolism of 2,4,5,2',4',5'-hexachlorobiphenyl (HCB) was investigated in vitro using liver microsomes of one male beagle dog after phenobarbital treatment.
2. Three major metabolites were isolated and identified as 3-hydroxy-2,4,5,2',4',5'-HCB, 2-hydroxy-4,5,2',4',5'-pentachlorobiphenyl (PenCB), and 2-hydroxy-3,4,5,2',4',5'-HCB, by comparison of g.l.c.-mass spectrometry and H-1-n.m.r. data with those of authentic samples.
3. 2-Hydroxy-3,4,5,2',4',5'-HCB was found as a metabolite of 2,4,5,2',4',5'-HCB for the first time using dog liver microsomes. Present results indicate that this metabolite and the dechlorinated PenCB are derived from a metabolic intermediate, namely, 2,3-epoxy-2,4,5,2',4',5'-HCB. 2,3-Epoxide formation is a new metabolic pathway of PCB.

DOI： 10.3109/00498259209053156

PURIFICATION AND CHARACTERIZATION OF 2 FORMS OF 2,3,4,7,8-PENTACHLORODIBENZOFURAN INDUCIBLE CYTOCHROME-P-450 IN HAMSTER LIVER Reviewed

N KOGA, N ARIYOSHI, H NAKASHIMA, H YOSHIMURA

JOURNAL OF BIOCHEMISTRY 107 ( 6 ) 826 - 833 1990.6

Language：English Publisher：JAPAN BIOCHEMICAL SOC

Golden SyrianハムスターにPenCDF (0.5 mg/kg)を膜腔内1回投与し,投与後5日目に屠殺し肝ミクロゾームを調製した.1.0%コール酸で可溶化後,ω-aminooctyl-Sepharose 4B, DE-52およびHydroxylapatiteの各カラムにて精製し,電気泳動的に単一で分子量52kDaのhamster P-450H(比含量12.8 nmol/mg protein)と分子量50 kDaのhamster P-450L(比含量7.73 nmol/mg protein)の2種を得た.いずれのP-450もCO-還元差スペクトルの吸収極大を446 nmに示し,それぞれ混合(高スピン+低スピン)型と低スピン型であった.なおhamster P-450H標品はhamster P-450Lの約27倍ものPenCDFを含有していた(0.107 nmol/nmol of P-450).両P-450に対する抗体を用いて未処理およびPenCDF処理ハムスター肝ミクロゾーム中の両P-450を定量したところ,hamster P-450Hは未処理ミクロゾーム全P-450量の約15%を占めること,およびPenCDF処理ではhamster P-450Hが60%,hamster P-450Lが30%を占めることが判明し,いずれの分子種もPenCDF誘導性であることが明らかとなった

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