2022/06/09 更新

写真a

ウノ ヤスヒロ
宇野 泰広
UNO Yasuhiro
所属
農水産獣医学域獣医学系 共同獣医学部 獣医学科 准教授
職名
准教授

学位

  • 博士(獣医学) ( 2008年3月   北海道大学 )

  • 獣医学士 ( 1994年3月   北海道大学 )

学歴

  • 北海道大学   獣医学科

    - 1994年3月

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    国名: 日本国

経歴

  • 鹿児島大学   農水産獣医学域獣医学系 共同獣医学部 獣医学科   准教授

    2019年9月 - 現在

  • 株式会社 新日本科学   薬物代謝分析センター   室長

    2006年4月 - 2019年8月

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    国名:日本国

  •   Research Assistant

    2003年4月 - 2006年3月

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    国名:日本国

  • 株式会社 新日本科学   研究員

    2001年11月 - 2003年3月

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    国名:日本国

  • University of Minnesota   Department of Animal Science   職員(技術系)

    2001年1月 - 2001年10月

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    国名:アメリカ合衆国

  • University of Minnesota   Molecular Veterinary Biosciences   大学院生(博士課程)

    1998年3月 - 2001年10月

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    国名:アメリカ合衆国

  • University of Massachusetts   Animal Science   大学院生(修士課程→博士課程)

    1994年9月 - 1998年2月

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    国名:アメリカ合衆国

▼全件表示

所属学協会

  • 日本獣医学会

    2006年4月 - 現在

留学歴

  • 1998年3月 - 2001年10月   University of Minnesota, St. Paul   Molecular Veterinary Bioscience

  • 1994年6月 - 1998年2月   University of Massachusetts, Amherst   Animal Science

取得資格

  • TOEIC(730点~)

  • 普通自動車免許(一種)

  • 獣医師

 

論文

  • Yasuhiro Uno, Hiroshi Yamazaki .  Systematic identification and characterization of cynomolgus macaque solute carrier transporters .  Drug Metabolism and Pharmacokinetics43   100437   2022年4月査読

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    担当区分:筆頭著者, 責任著者   記述言語:英語  

  • Yasuhiro Uno, Shotaro Uehara, and Hiroshi Yamazaki .  Drug-oxidizing and conjugating non-cytochrome P450 (non-P450) enzymes in cynomolgus monkeys and common marmosets as preclinical models for humans .  Biochemical Pharmacology197   114887   2022年3月査読

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    担当区分:筆頭著者   記述言語:英語  

  • Yasuhiro Uno, Hiroshi Yamazaki .  Cloning and tissue expression of ATP-binding cassette transporters in cynomolgus macaques .  Drug Metabolism and Pharmacokinetics42   100431   2022年2月査読

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    担当区分:筆頭著者, 責任著者   記述言語:英語  

  • Uno Y, Shimizu M, Ogawa Y, Makiguchi M, Kawaguchi H, Yamato O, Ishizuka M, and Yamazaki H .  Molecular and functional characterization of flavin-containing monooxygenases in pigs, dogs, and cats .  Biochem Pharmacolin press   2022年査読

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    担当区分:筆頭著者, 責任著者  

  • Uno Y, Murayama N, Ijiri M, Kawaguchi H, Yamato O, Shiraishi M, Asano A, Teraoka H, Mizukawa H, Nakayama SM, Ikenaka Y, Ishizuka M, and Yamazaki H .  Cytochrome P450 2J (CYP2J) genes in dogs, cats, and pigs are expressed and encode functional drug-metabolizing enzymes .  Drug Metab Disposin press   2022年

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    担当区分:筆頭著者, 責任著者  

  • Uno Y, Uehara S, Ijiri M, Kawaguchi H, Asano A, Shiraishi M, Banju K, Murayama N, and Yamazaki H .  Molecular and functional characterization of N-acetyltransferases in common marmosets and pigs .  Drug Metab Disposin press   2022年

  • Uno Y, Jikuya S, Noda Y, Murayama N, and Yamazaki H .  Comprehensive investigation of dog cytochromes P450 3A (CYP3A) revealed functional role of newly identified CYP3A98 in small intestine .  Drug Metab Disposin press   2022年

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    担当区分:筆頭著者, 責任著者  

  • Shiori Honda, Tatsuki Fukami, Keiya Hirosawa, Takuya Tsujiguchi, Yongjie Zhang, Masataka Nakano, Shotaro Uehara, Yasuhiro Uno, Hiroshi Yamazaki, and Miki Nakajima .  Differences in hydrolase activities in the liver and small intestine between marmosets and humans .  Drug Metab Dispos49 ( 9 ) 718 - 728   2021年9月査読

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Shotaro Uehara, Yasuhiro Uno, Makiko Shimizu, and Hiroshi Yamazaki .  Cloning, sequence analysis, and tissue expression of marmoset paraoxonase 1 .  Drug Metab Pharmacokinet39 ( 2021 ) 100398   2021年8月査読

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    記述言語:英語  

  • Yurie Okawa, Sakae Kohara, Asako Uchiyama, Hiroshi Yamazaki, Yasuhiro Uno .  Evaluation of domain of unknown function 1220 (DUF1220) for detection of human genome by quantitative polymerase chain reaction: Potential use in assessing the biodistribution of transplanted therapeutic human cells .  Drug Metab Pharmacokinet38 ( 2021 ) 100366   2021年6月査読

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    担当区分:筆頭著者, 最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno, Shotaro Uehara, Norie Murayama, Hiroshi Yamazaki .  Genetic variants of aldehyde oxidase (AOX) 1 in cynomolgus and rhesus macaques .  Xenobiotica51 ( 4 ) 494 - 499   2021年4月査読

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    担当区分:筆頭著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno, Takahiro Mikami, Yasuko Tsukazaki, Yasuharu Nakanishi, Norie Murayama, Shinichi Ikushiro, Hideshi Tsusaki, Hiroshi Yamazaki .  Genetic variants of UDP-glucuronosyltransferases 1A1, 1A6, and 1A9 in cynomolgus and rhesus macaques .  Xenobiotica51 ( 1 ) 115 - 121   2021年1月査読

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    担当区分:筆頭著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Makiko Shimizu, Yasuhiro Uno, Masahiro Utoh, and Hiroshi Yamazaki .  Trimethylamine N-oxygenation in cynomolgus macaques genotyped for flavin-containing monooxygenase 3 (FMO3) .  Drug Metab Pharmacokinet35 ( 6 ) 571 - 573   2020年12月査読

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    担当区分:責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Shotaro Uehara, Yasuhiro Uno, Hiroshi Yamazaki .  Molecular cloning and tissue distribution of marmoset thiopurine S-methyltransferase. .  Drug Metab Pharmacokinet35 ( 5 ) 475 - 478   2020年10月査読

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno, Shotaro Uehara, Norie Murayama, Makiko Shimizu, Hiroshi Yamazaki .  Expression of functional sulfotransferases (SULT) 1A1, 1A3, 1B1, 1C2, 1E1, and 2A1 in common marmosets .  Biochem Pharmacol180 ( 2020 ) 114189   2020年10月査読

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    担当区分:筆頭著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Shotaro Uehara, Yasuhiro Uno, and Hiroshi Yamazaki .  Molecular cloning, sequence analysis, and tissue distribution of marmoset monoamine oxidases A and B .  Drug Metab Pharmacokinet35 ( 5 ) 479 - 482   2020年10月査読

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno, Hiroshi Yamazaki .  Molecular characterization of UDP-glucuronosyltransferases 3A and 8A in cynomolgus macaques .  Drug Metab Pharmacokinet35 ( 4 ) 397 - 400   2020年8月査読

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    担当区分:筆頭著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuharu Nakanishi, Yasuhiro Uno, Hiroshi Yamazaki .  Regional distributions of UDP-glucuronosyltransferase activities toward estradiol and serotonin in the liver and small intestine of cynomolgus macaques .  Drug Metab Pharmacokinet35 ( 4 ) 401 - 404   2020年8月査読

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    担当区分:責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno, Norie Murayama, and Hiroshi Yamazaki .  Genetic variants of N-acetyltransferases 1 and 2 (NAT1 and NAT2) in cynomolgus and rhesus macaques .  Biochem Pharmacol177 ( 2020 ) 113996   2020年7月査読

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    担当区分:筆頭著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno, Hiroshi Yamazaki .  Expression levels of microRNAs that are potential cytochrome P450 regulators in cynomolgus macaques .  Xenobiotica50 ( 7 ) 747 - 752   2020年7月査読

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    担当区分:筆頭著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno, Norie Murayama, and Hiroshi Yamazaki .  Interleukin-1β and tumor necrosis factor-α affect cytochrome P450 expression in cynomolgus macaque hepatocytes .  Drug Metab Pharmacokinet35 ( 3 ) 341 - 343   2020年6月査読

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno, Shotaro Uehara, Saki Tanaka, Norie Murayama, and Hiroshi Yamazaki .  Systematic characterization of glutathione S-transferases in common marmosets .  Biochem Pharmacol174 ( 2020 ) 113835   2020年4月査読

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno and Hiroshi Yamazaki .  mRNA levels of drug-metabolizing enzymes in 11 brain regions of cynomolgus macaques .  Drug Metab Pharmacokinet35 ( 2 ) 248 - 252   2020年4月査読

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Shotaro Uehara, Yasuhiro Uno, Takashi Inoue, Erika Sasaki, and Hiroshi Yamazaki .  Cloning and tissue expression of cytochrome P450 2S1, 4V2, 7A1, 7B1, 8B1, 24A1, 26A1, 26C1, 27A1, 39A1, and 51A1 in marmosets .  Drug Metab Pharmacokinet35 ( 2 ) 244 - 247   2020年4月査読

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Kayoko Ohura, Yoshiyuki Igawa, Maori Tanaka, Kana Matsumoto, Akiko Kasahara, Naoya Wada, Kazuishi Kubota, Yasuhiro Uno, and Teruko Imai .  Identification and characterization of a new carboxylesterase 2 isozyme, mfCES2C, in small intestine of cynomolgus monkey .  Drug Metab Dispos 48 ( 3 ) 146 - 152   2020年3月査読

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno, Shotaro Uehara, Takashi Inoue, Shu Kawamura, Norie Murayama, Miyu Nishikawa, Shinichi Ikushiro, Erika Sasaki, and Hiroshi Yamazaki .  Molecular characterization of functional UDP-glucuronosyltransferases 1A and 2B in common marmosets .  Biochem Pharmacol170 ( 2020 ) 113748   2020年2月査読

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Shotaro Uehara , Yasuhiro Uno , and Hiroshi Yamazaki .  The marmoset cytochrome P450 superfamily: sequence/phylogenetic analyses, genomic structure, and catalytic function .  Biochem Pharmacol171 ( 2020 ) 113721   2020年1月査読

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Uno Yasuhiro, Murayama Norie, Yamazaki Hiroshi .  Molecular and functional characterization of cytosolic sulfotransferases in cynomolgus macaque. .  Biochem Pharmacol166   153 - 162   2019年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cytosolic sulfotransferases (SULTs), drug-metabolizing enzymes essential for the metabolism of endogenous biochemicals and foreign compounds, have been characterized in humans, but remain to be investigated in cynomolgus macaques, important species in drug metabolism studies. In this study, based on the genome data, cynomolgus SULT1A1, SULT1A3, SULT1B1, SULT1C2v1, SULT1C2v2, SULT1C4, SULT1E1, and SULT2A1 cDNAs were isolated and characterized. Among these, cynomolgus SULT1C2v2 was highly homologous to human SULT1C2P1 (pseudogene). These cynomolgus SULT cDNAs had high sequence identities (95-97%) to, and closely clustered with their human orthologs in a phylogenetic tree. Gene structure and genomic organization of each cynomolgus SULT were similar to those of the human ortholog. Among the 10 tissue types analyzed, cynomolgus SULTs showed distinct expression patterns similar to human SULTs; more specifically, mRNA was most abundantly expressed in livers (SULT1A1, SULT1C2v2, SULT1C4, and SULT2A1), jejunum (SULT1A3, SULT1B1, and SULT1E1), or kidneys (SULT1C2v1). The most abundant SULT mRNA was SULT2A1, SULT1E1, and SULT1C4 found in livers, jejunum, and kidneys, respectively. Recombinant cynomolgus SULT1A1, SULT1A3, SULT1B1, SULT1C2v1, SULT1C2v2, SULT1C4, SULT1E1, and SULT2A1 in bacterial cytosolic fractions mediated 3'-phosphoadenosine-5'-phosphosulfate-dependent sulfate conjugations of typical human SULT substrates, 1-naphthol, p-nitrophenol, dopamine, dehydroepiandrosterone, and estradiol. Taken together, these results suggest molecular and functional similarities of SULTs between cynomolgus macaques and humans.

    DOI: 10.1016/j.bcp.2019.05.018

    PubMed

  • Uno Yasuhiro, Murayama Norie, Tamura Kazuaki, Yamazaki Hiroshi .  Functionally relevant genetic variants of glutathione S-transferase GSTM5 in cynomolgus and rhesus macaques. .  Xenobiotica49 ( 8 ) 995 - 1000   2019年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Glutathione S-transferase (GST) is a family of enzymes important for conjugation with glutathione of endogenous and exogenous compounds. Human GSTM1 null allele is associated with toxicity and cancers. Cynomolgus and rhesus macaques have molecular and enzymatic similarities of GSTs to humans; however, genetic variants have not been investigated. In macaques, instead of pseudogenized GSTM1, GSTM5 is a predominant GSTM isoform. In this study, re-sequencing of GSTM5 in 64 cynomolgus and 31 rhesus macaques found 6 non-synonymous variants, and 1 variant (IVS5 + 1) causing exon skip. Of these 7 variants, 3 and 1 were found only in Indochinese and Indonesian cynomolgus macaques, respectively. Cynomolgus GSTM5-mediated styrene 7,8-oxide and trans-stilbene oxide conjugation activities correlated with GSTM protein levels immunochemically quantified in cynomolgus liver samples. Using recombinant GSTM5 proteins, 4 of the 6 non-synonymous variants including E29Q, L96R, M166V and S201N showed substantially lower metabolic activities. Moreover, a homozygote for E29Q and heterozygotes for S201N or IVS5 + 1 showed significantly lower conjugation activities in liver cytosolic fractions as compared with wild-type animals. Therefore, the present results suggest that inter-animal variability of GST-dependent drug metabolism is at least partly accounted for by GSTM5 variants in cynomolgus and rhesus macaques as pre-clinical animal models.

    DOI: 10.1080/00498254.2018.1524187

    PubMed

  • Sugiyama Souta, Uno Yasuhiro, Amano Tomoko, Kitazawa Takio, Teraoka Hiroki .  Genetic diversity of cytochrome P450 1A2 with different metabolic activities in domestic cats. .  J Vet Med Sci81 ( 7 ) 980 - 982   2019年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Knowledge of genetic polymorphisms of metabolizing enzymes of medical drugs and xenobiotics including cytochrome P450 (CYP) is very limited in cats. We investigated polymorphisms in CYP1A2, one of the major CYP isoforms in the feline liver. Wild-type and three non-synonymous polymorphic variants, but no synonymous variant, were identified in feline CYP1A2 in 50 alleles of domestic cats in Japan. Metabolic parameters, Km and Vmax, of ethoxyresorufin hydroxylation by CYP1A2 were shown to range within two times for identified non-synonymous variants by using a heterologous coexpression system. The results confirmed the polymorphic nature of CYP1A2 as a basis for effective application of medicines and prevention of adverse reactions in the treatment of domestic cats as well as for hereditary disorders.

    DOI: 10.1292/jvms.19-0106

    PubMed

  • Sugiyama Souta, Uno Yasuhiro, Amano Tomoko, Kitazawa Takio, Teraoka Hiroki .  Genetic diversity of cytochrome P450 2A with different metabolic activities in domestic cats. .  J Vet Med Sci81 ( 7 ) 983 - 985   2019年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Knowledge of genetic polymorphisms of cytochrome P450 (CYP), the most important xenobiotic metabolizing enzyme, is very limited in cats. Preliminarily, we investigated genetic polymorphisms in CYP2A13, one of the major CYP isoforms in the liver and lung. Four synonymous and three non-synonymous polymorphic variants were identified in feline CYP2A13 in domestic cats in Japan, without an obvious major type. Metabolic parameters, Km and Vmax, of coumarin hydroxylation of CYP2A13 were shown to range within two times for the identified non-synonymous polymorphic variants by using heterologous coexpression system in Escherichia coli. The results confirmed the polymorphic nature of CYP2A13 as a basis for effective application of medicines and prevention of adverse reactions in treatment of domestic cats.

    DOI: 10.1292/jvms.19-0107

    PubMed

  • Ono Yuka, Sugiyama Souta, Matsushita Mayu, Kitazawa Takio, Amano Tomoko, Uno Yasuhiro, Ikushiro Shinichi, Teraoka Hiroki .  Limited expression of functional cytochrome p450 2c subtypes in the liver and small intestine of domestic cats. .  Xenobiotica49 ( 6 ) 627 - 635   2019年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Compared to information for herbivores and omnivores, knowledge on xenobiotic metabolism in carnivores is limited. The cytochrome P450 2C (CYP2C) subfamily is recognized as one of the most important CYP groups in human and dog. We identified and characterized CYP2C isoforms and variants in cat, which is an obligate carnivore. 2. Quantitative RT-PCR and immunoblot analyses were carried out to evaluate the expression of CYP2C in the liver and small intestine. A functional CYP2C isoform was heterologously expressed in yeast microsomes to determine the enzymatic activity. 3. Cat had two CYP2C genes, 21 and 41, in the genome; however, CYP2C21P was a pseudogene that had many stop codons. Three splicing variants of CYP2C41 were identified (v1-v3), but only one of them (v1) showed a complete deduced amino acid sequence as CYP2C protein. Transcripts of feline CYP2C41v1 were detected but the amounts were negligible or very small in the liver and small intestine. Immunoreactivity to an antihuman CYP2C antibody was confirmed in the recombinant feline CYP2C41v1 but not in the feline liver. 4. Recombinant feline CYP2C41v1 metabolized several substrates, including dibenzylfluorescein that is specific to human CYP2C. 5. The results suggest a limited role of functional CYP2C isoforms in xenobiotic metabolism in cat.

    DOI: 10.1080/00498254.2018.1483543

    PubMed

  • Uno Yasuhiro, Takahira Rika, Murayama Norie, Onozeki Shunsuke, Kawamura Shu, Uehara Shotaro, Ikenaka Yoshinori, Ishizuka Mayumi, Ikushiro Shinichi, Yamazaki Hiroshi .  Functional and molecular characterization of UDP-glucuronosyltransferase 2 family in cynomolgus macaques. .  Biochem Pharmacol163   335 - 344   2019年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    UDP-glucuronosyltransferases (UGTs) are essential enzymes metabolizing endogenous and exogenous chemicals. However, characteristics of UGTs have not been fully investigated in molecular levels of cynomolgus macaques, one of non-human primates widely used in preclinical drug metabolism studies. In this study, three UGT2A cDNAs (UGT2A1, 2A2, and 2A3) were isolated and characterized along with seven UGT2Bs previously identified in cynomolgus macaques. Several transcript variants were found in cynomolgus UGT2A1 and UGT2A2, like human orthologs. Cynomolgus UGT2A and UGT2B amino acid sequences were highly identical (87-96%) to their human counterparts. By phylogenetic analysis, all these cynomolgus UGT2s were more closely clustered with their human homologs than with dog, rat, or mouse UGT2s. Especially, UGT2As showed orthologous relationships between humans and cynomolgus macaques. All the cynomolgus UGT2 mRNAs were expressed in livers, jejunum, and/or kidneys abundantly, except that UGT2A1 and UGT2A2 mRNAs were predominantly expressed in nasal mucosa, like human UGT2s. UGT2A and UGT2B genes together form a gene cluster in the cynomolgus and human genome. Among the seven cynomolgus UGT2Bs heterologously expressed in yeast, UGT2B9 and UGT2B30 showed activities in estradiol 17-O-glucuronidation and morphine 3-O-glucuronidation but did not show activities in estradiol 3-O-glucuronidation, similar to human UGT2Bs. In liver microsomes, cynomolgus macaques showed higher estradiol 17-O-glucuronidase and morphine 3-O-glucuronidase activities than humans, suggesting functional activities of the responsible UGT2B enzymes in cynomolgus macaques. Therefore, cynomolgus UGT2s had overall molecular similarities to human UGT2s, but also showed some differences in UGT2B enzyme properties.

    DOI: 10.1016/j.bcp.2019.03.002

    PubMed

  • Sugiyama Souta, Uno Yasuhiro, Amano Tomoko, Kitazawa Takio, Teraoka Hiroki .  Genetic diversity of cytochrome P450 3A with different metabolic activity in domestic cats. .  J Vet Med Sci81 ( 4 ) 598 - 600   2019年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Knowledge on genetic polymorphisms of metabolising enzymes including cytochrome P450 (CYP) is very limited in cats. We investigated polymorphisms in CYP3A131, one of the major CYP isoforms in the feline liver and small intestine. Eight non-synonymous variants and one synonymous variant of feline CYP3A131 were identified in 29 cats. A major non-synonymous type was not observed. Metabolic parameters (Km and Vmax) of dibenzylfluorescein hydroxylation were ranged within about 2 times for the identified non-synonymous variants by using a heterologous coexpression system of CYP3A131 and feline cytochrome P450 reductase in Escherichia coli. The results confirmed the polymorphic nature of CYP3A131 as a basis for effective application of medicines and prevention of adverse reactions in the treatment of domestic cats.

    DOI: 10.1292/jvms.18-0692

    PubMed

  • Oshio Toru, Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Marmoset cytochrome P450 2B6, a propofol hydroxylase expressed in liver. .  Xenobiotica49 ( 3 ) 265 - 269   2019年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The common marmoset (Callithrix jacchus) is a useful experimental animal to evaluate the pharmacokinetics of drug candidates. Cytochrome P450 (P450) 2B enzyme in marmoset livers has been identified; however, only limited information on the enzymatic properties and distribution has been available. Marmoset P450 2B6 amino acids showed high sequence identities (>86%) with those of primates including humans and cynomolgus monkeys. Phylogenetic analysis using amino acid sequences indicated that marmoset P450 2B6 was closer to human and cynomolgus monkey P450 2B6 than to P450 2B orthologs of other species, including pigs, dogs, rabbits and rodents. Quantitative polymerase chain reaction analysis using specific primers showed P450 2B6 mRNA predominantly expressed in livers among the five marmoset tissues, similar to those of humans and cynomolgus monkeys. Marmoset P450 2B6 heterologously expressed in Escherichia coli membranes oxidized 7-ethoxycoumarin, pentoxyresorufin, propofol and testosterone, at roughly similar rates to those of humans and/or cynomolgus monkeys. A high capacity of marmoset P450 2B6 with propofol 4-hydroxylation (at low ionic strength conditions) with a low Km value was relatively comparable to that for marmoset livers. These results collectively indicated a high propofol 4-hydroxylation activity of P450 2B6 expressed in marmoset livers.

    DOI: 10.1080/00498254.2018.1439204

    PubMed

  • Uno Yasuhiro, Shimizu Makiko, Yoda Hiromi, Yamazaki Hiroshi .  Expression and metabolic activity of flavin-containing monooxygenase 1 in cynomolgus macaque kidney. .  J Med Primatol48 ( 1 ) 51 - 53   2019年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Flavin-containing monooxygenase 1 (FMO1) largely remains to be characterized in cynomolgus macaque kidney. Immunoblotting showed expression of cynomolgus FMO1 in kidneys where activities of FMO1 (benzydamine N-oxygenation) were detected. No sex differences were observed in their contents or activities. These results suggest the functional role of cynomolgus FMO1 in kidney.

    DOI: 10.1111/jmp.12385

    PubMed

  • Takahashi Tsuyoshi, Uno Yasuhiro, Yamazaki Hiroshi, Kume Toshiyuki .  Functional characterization for polymorphic organic anion transporting polypeptides (OATP/SLCO1B1, 1B3, 2B1) of monkeys recombinantly expressed with various OATP probes. .  Biopharm Drug Dispos40 ( 2 ) 62 - 69   2019年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The hepatic uptake of clinical drugs mediated by human hepatic organic anion transporting polypeptides (OATP/SLCO) has been reported extensively. In this study, hepatic uptake by recombinantly expressed monkey OATP1B1, OATP1B3 and OATP2B1 was investigated using three human OATP1B1 and OATP1B3 substrates (pitavastatin, pravastatin and rosuvastatin) and one OATP1B3 substrate (telmisartan), as the governmental drug interaction guidelines recommend, and seven reported clinical drugs. The uptake of known human probes into recombinant OATP- expressing cells was significantly greater than that of mock cells. Consequently, pitavastatin, pravastatin and rosuvastatin were suggested to be substrates of recombinant monkey OATP1B1 and OATP1B3, and telmisartan was suggested to be a substrate of recombinant monkey OATP1B3, in a manner similar to human OATPs. In contrast, atorvastatin, bosentan, etoposide, fexofenadine, fluvastatin, glibenclamide and simeprevir were broadly transported by recombinant monkey OATP1B1, OATP1B3 and OATP2B1. Furthermore, some of the 16 non-synonymous monkey OATP1B1 variants found in 64 cynomolgus and 32 rhesus monkeys mediated up to a 1.6-fold [(3) H]pitavastatin uptake (with low Michaelis constant values) in comparison with the wild type under the present conditions. Despite sequences of monkey recombinant OATPs not being totally reflective of those of human OATPs, our results collectively suggested that OATP1B1, OATP1B3 or OATP2B1 in monkeys could mediate roughly a similar hepatic uptake of various OATP probes. Recombinant monkey OATPs would be good experimental tools for in vitro hepatic uptake in cell systems.

    DOI: 10.1002/bdd.2171

    PubMed

  • Iwasaki Kazuhide, Uno Yasuhiro, Utoh Masahiro, Yamazaki Hiroshi .  Importance of cynomolgus monkeys in development of monoclonal antibody drugs. .  Drug Metab Pharmacokinet34 ( 1 ) 55 - 63   2019年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Animal species used in the preclinical studies for development of monoclonal antibody (mAb) drugs are surveyed in this review. Relevant animal species for preclinical studies of mAb candidates are those express desired epitope of mAb candidates. Cynomolgus monkeys cross-react with mAb drugs much higher than other animal species commonly used in preclinical studies such as absorption, distribution, metabolism and excretion (ADME), efficacy, and toxicity studies, for development of new drugs. Moreover, plasma exposure of the mAb drugs in humans is predicted well from the exposure in the monkeys, and the placental transfer of immunoglobulin G (IgG, all the mAb drugs contain IgG) from mother to fetus is similar between humans and the monkeys from a viewpoint of time course and plasma level of IgG transferred. These observed findings indicate that the monkeys are the most suitable animal species used in the ADME and toxicity studies for development of new mAb drugs.

    DOI: 10.1016/j.dmpk.2018.02.003

    PubMed

  • Uno Yasuhiro, Igawa Yoshiyuki, Tanaka Maori, Ohura Kayoko, Hosokawa Masakiyo, Imai Teruko .  Analysis of carboxylesterase 2 transcript variants in cynomolgus macaque liver. .  Xenobiotica49 ( 2 ) 247 - 255   2019年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Carboxylesterase (CES) is important for the detoxification of a wide range of drugs and xenobiotics. In this study, the hepatic level of CES2 mRNA was examined in cynomolgus macaques used widely in preclinical studies for drug metabolism. Three CES2 mRNAs were present in cynomolgus macaque liver. The mRNA level was highest for cynomolgus CES2A (formerly CES2v3), much lower for cynomolgus CES2B (formerly CES2v1) and extremely low for cynomolgus CES2C (formerly CES2v2). Most various transcript variants produced from cynomolgus CES2B gene did not contain a complete coding region. Thus, CES2A is the major CES2 enzyme in cynomolgus liver. A new transcript variant of CES2A, CES2Av2, was identified. CES2Av2 contained exon 3 region different from wild-type (CES2Av1). In cynomolgus macaques expressing only CES2Av2 transcript, CES2A contained the sequence of CES2B in exon 3 and vicinity, probably due to gene conversion. On genotyping, this CES2Av2 allele was prevalent in Indochinese cynomolgus macaques, but not in Indonesian cynomolgus or rhesus macaques. CES2Av2 recombinant protein showed similar activity to CES2Av1 protein for several substrates. It is concluded that CES2A is the major cynomolgus hepatic CES2, and new transcript variant, CES2Av2, has similar functions to CES2Av1.

    DOI: 10.1080/00498254.2018.1435927

    PubMed

  • Uno Yasuhiro, Shimizu Makiko, Yoda Hiromi, Origuchi Yumi, Yamazaki Hiroshi .  Non-synonymous genetic variants of flavin-containing monooxygenase 3 (FMO3) in cynomolgus macaques. .  Drug Metab Pharmacokinet34 ( 1 ) 104 - 107   2019年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Polymorphic human flavin-containing monooxygenase (FMO) 3 is an important drug-metabolizing enzyme for nitrogen- or sulfur-containing compounds. Cynomolgus macaques, a non-human primate species widely used in drug metabolism studies, have corresponding FMO3 molecular and enzymatic similarities to humans; however, genetic polymorphisms have not been investigated in macaques. In this study, re-sequencing of FMO3 in 64 cynomolgus and 32 rhesus macaques found a total of 18 non-synonymous variants. Nine variants were unique to cynomolgus macaques, of which 4 (including Q506K) were found only in Indochinese, 4 (including V299I, E348H, and G530A) only in Indonesian lineages, and one was common. Other five variants (including S504T at >10% allele frequencies) were unique to rhesus macaques. By functional characterization using cynomolgus FMO3 proteins heterologously expressed in Escherichia coli, FMO3 R509H variant appeared to suppress methimazole and benzydamine S- or N-oxygenations. Seven variants showed substantially lower benzydamine N-oxygenation as compared with wild-type FMO3 protein. Further analysis indicated that two of these variants, FMO3 G530A and R417H, showed significantly lower benzydamine N-oxygenation in liver microsomes of the homozygotes as compared with wild-type animals. Therefore, inter-animal variability of FMO3-dependent drug metabolism is at least partly accounted for by genetic polymorphisms in cynomolgus and rhesus macaques, similar to humans.

    DOI: 10.1016/j.dmpk.2018.09.001

    PubMed

  • Uehara Shotaro, Oshio Toru, Nakanishi Kazuyuki, Tomioka Etsuko, Suzuki Miyu, Inoue Takashi, Uno Yasuhiro, Sasaki Erika, Yamazaki Hiroshi .  Survey of Drug Oxidation Activities in Hepatic and Intestinal Microsomes of Individual Common Marmosets, a New Nonhuman Primate Animal Model. .  Curr Drug Metab20 ( 2 ) 103 - 113   2019年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Common marmosets (Callithrix jacchus) are potentially useful nonhuman primate models for preclinical studies. Information for major drug-metabolizing cytochrome P450 (P450) enzymes is now available that supports the use of this primate species as an animal model for drug development. Here, we collect and provide an overview of information on the activities of common marmoset hepatic and intestinal microsomes with respect to 28 typical human P450 probe oxidations. RESULTS: Marmoset P450 2D6/8-dependent R-metoprolol O-demethylation activities in hepatic microsomes were significantly correlated with those of midazolam 1'- and 4-hydroxylations, testosterone 6beta-hydroxylation, and progesterone 6beta-hydroxylation, which are probe reactions for marmoset P450 3A4/5/90. In marmosets, the oxidation activities of hepatic microsomes and intestinal microsomes were roughly comparable for midazolam and terfenadine. Overall, multiple forms of marmoset P450 enzymes in livers and intestines had generally similar substrate recognition functionalities to those of human and/or cynomolgus monkey P450 enzymes. CONCLUSION: The marmoset could be a model animal for humans with respect to the first-pass extraction of terfenadine and related substrates. These findings provide a foundation for understanding individual pharmacokinetic and toxicological results in nonhuman primates as preclinical models and will help to further support understanding of the molecular mechanisms of human P450 function.

    DOI: 10.2174/1389200219666181003143312

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Murayama Norie, Yamazaki Hiroshi .  Cytochrome P450 1A1, 2C9, 2C19, and 3A4 Polymorphisms Account for Interindividual Variability of Toxicological Drug Metabolism in Cynomolgus Macaques. .  Chem Res Toxicol31 ( 12 ) 1373 - 1381   2018年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cytochromes P450 (P450s) and their genetic variants in humans are important drug-metabolizing enzymes partly accounting for interindividual variations in drug metabolism and toxicity. However, these genetic variants in P450s have not been fully investigated in cynomolgus macaques, a nonhuman primate species widely used in toxicological studies. In this study, genetic variants found in cynomolgus CYP1A1, CYP2C9 (formerly CYP2C43), CYP2C19 (CYP2C75), and CYP3A4 (CYP3A8) were assessed on functional importance. Resequencing of CYP1A1 in cynomolgus macaques found 18 nonsynonymous variants, of which M121I and V382I were located in SRSs, domains potentially important for P450 function. By further analyzing these two variants, V382I was significantly associated with lower drug-metabolizing activities in the liver for the heterozygotes than the wild types. Similarly, the heterozygotes or homozygotes of CYP2C9 variants (A82V and H344R) and CYP2C19 variant (A490V) showed significantly lower drug-metabolizing activities in the liver than the wild types. Moreover, the homozygotes of CYP3A4 variant (S437N) showed significantly higher activities than the wild type in the liver. Kinetic analyses using recombinant proteins revealed that CYP2C9 variants (A82V and H344R) showed substantially lower Ks values than the wild type, although CYP1A1 variant (V382I) showed kinetic parameters similar to the wild type. Likewise, CYP2C19 variant (A490V) showed substantially a lower Vmax/ Km value than the wild type, whereas CYP3A4 variant (S437N) showed a higher Vmax/ Km value than the wild type. These results suggest the toxicologically functional importance of CYP2C9 variants (A82V and H344R), CYP2C19 variant (A490V), and CYP3A4 variant (S437N) for hepatic drug metabolism in cynomolgus macaques.

    DOI: 10.1021/acs.chemrestox.8b00257

    PubMed

  • Uno Yasuhiro, Murayama Norie, Yamazaki Hiroshi .  Molecular and Functional Characterization of N-Acetyltransferases NAT1 and NAT2 in Cynomolgus Macaque. .  Chem Res Toxicol31 ( 11 ) 1269 - 1276   2018年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Arylamine N-acetyltransferases (NATs) are drug-metabolizing enzymes essential for the metabolism of endogenous substrates and xenobiotics, and their molecular characteristics have been extensively investigated in humans, but not in cynomolgus macaques, nonhuman primate species important for drug metabolism studies. In this study, cynomolgus NAT1 and NAT2 cDNAs were isolated from livers. NAT1 and NAT2 were characterized by molecular analyses and drug-metabolizing assays. A total of 9 transcript variants were found for cynomolgus NAT1, similar to human NAT1, and contained 1-4 exons with the coding region largely conserved with human NAT1. Genomic organization was similar between cynomolgus macaques and humans. Cynomolgus NAT1 and NAT2 amino acid sequences showed high sequence homology (95% and 89%, respectively) and showed close relationships with human NAT1 and NAT2 in a phylogenetic tree. Cynomolgus NAT2 mRNA was predominantly expressed in liver among the 10 different tissues analyzed, followed by kidney and jejunum. In contrast, cynomolgus NAT1 mRNA showed more ubiquitous expression with relatively more abundant expression in liver, kidney, and jejunum, along with testis. Metabolic assays using recombinant proteins showed that cynomolgus NAT1 and NAT2 metabolized human NAT substrates, including p-aminobenzoic acid, sulfamethazine, isoniazid, and 2-aminofluorene. Interestingly, p-aminobenzoic acid and isoniazid were largely metabolized by NAT1 and NAT2, respectively, in cynomolgus macaques and humans; sulfamethazine, a human NAT2 substrate, was metabolized by both NAT enzymes in cynomolgus macaques. These results suggest molecular and enzymatic similarities of NAT1 and NAT2 between cynomolgus macaques and humans, despite some small differences in substrate specificity of the enzymes.

    DOI: 10.1021/acs.chemrestox.8b00236

    PubMed

  • Uno Yasuhiro, Murayama Norie, Kato Masami, Tanaka Saki, Ohkoshi Tomoko, Yamazaki Hiroshi .  Genetic Variants of Glutathione S-Transferase GSTT1 and GSTT2 in Cynomolgus Macaques: Identification of GSTT Substrates and Functionally Relevant Alleles. .  Chem Res Toxicol31 ( 10 ) 1086 - 1091   2018年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Glutathione S-transferase (GST) is a family of important drug- metabolizing enzymes, conjugating endogenous and exogenous compounds. Genetic polymorphisms result in the inter-individual variability of GST activity in humans. Especially, human GSTT1 and GSTT2 null alleles are associated with toxicity and various cancers derived from chemicals. Cynomolgus macaque, a nonhuman primate species widely used in drug metabolism studies, has molecular and enzymatic similarities of GSTs to the human orthologs; however, genetic polymorphisms have not been investigated in this species. In this study, resequencing of GSTT1 and GSTT2 in 64 cynomolgus and 32 rhesus macaques found 15 nonsynonymous variants and 1 nonsense variant for GSTT1 and 15 nonsynonymous variants for GSTT2. Some of these GSTT variants were distributed differently in Indochinese and Indonesian cynomolgus macaques and rhesus macaques. For analysis of functional relevance of the GSTT variants, 1-iodohexane and dibromomethane were determined to be suitable substrates for cynomolgus GSTT1 and GSTT2. However, the conjugation activities were roughly correlated with GSTT protein levels immunochemically quantified in cynomolgus liver samples with no statistical significances, implying the contributions of the GST genetic variants. Among the GSTT1 variants identified, the animals carrying R76C and D125G mutations showed lower conjugation activities toward dibromomethane than those of the wild-type in liver cytosolic fractions. Moreover, the recombinant R76C/D125G and D125G GSTT variant proteins showed significantly lower 1-iodohexane or dibromomethane conjugation activities than those of the wild-type protein. Therefore, inter-animal variability of GSTT-dependent drug metabolism is at least partly accounted for by GSTT1 and possibly GSTT2 variants in cynomolgus and rhesus macaques.

    DOI: 10.1021/acs.chemrestox.8b00198

    PubMed

  • Kusama Takashi, Toda Akiko, Shimizu Makiko, Uehara Shotaro, Inoue Takashi, Uno Yasuhiro, Utoh Masahiro, Sasaki Erika, Yamazaki Hiroshi .  Association with polymorphic marmoset cytochrome P450 2C19 of in vivo hepatic clearances of chirally separated R-omeprazole and S-warfarin using individual marmoset physiologically based pharmacokinetic models. .  Xenobiotica48 ( 10 ) 1072 - 1077   2018年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Simulated clearances of R-warfarin and efavirenz were recently reported for individual cynomolgus monkeys genotyped for cytochrome P450 2C19 and 2C9, respectively. To expand and verify this modeling procedure, simulations of R/S-omeprazole and R/S-warfarin clearances after oral administrations in individual marmosets were performed using individual simplified physiologically based pharmacokinetic (PBPK) modeling consisting of gut, liver and central compartments. 2. Pharmacokinetics of R/S-omeprazole were chirally determined using the previously reported plasma microsamples in this study. The areas under the plasma concentration/time curves (AUC) of R-omeprazole and S-warfarin, but not S-omeprazole and R-warfarin, after oral administrations in the P450 2C19 homozygous mutant group were significantly higher than those in the wild- type group. These modeled hepatic intrinsic clearances were also significantly associated with the marmoset P450 2C19 genotypes. Other parameter values, e.g. absorption rate constants or systemic circulation volumes, were not likely determining factors. 3. The reported individual AUC values measured in 4-6 marmosets after oral R-omeprazole and S-warfarin administrations were significantly correlated with the AUC values predicted using the PBPK models after virtual administrations. 4. This study indicates that clearances of R-omeprazole, S-warfarin and related medicines associated with polymorphic P450 2C19 in individual marmosets can be simulated using simplified individual PBPK models.

    DOI: 10.1080/00498254.2017.1393121

    PubMed

  • Uno Yasuhiro, Takahira Rika, Murayama Norie, Ishii Yu, Ikenaka Yoshinori, Ishizuka Mayumi, Yamazaki Hiroshi, Ikushiro Shinichi .  Molecular and functional characterization of UDP-glucuronosyltransferase 1A in cynomolgus macaques. .  Biochem Pharmacol155   172 - 181   2018年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    UDP-glucuronosyltransferases (UGTs) are drug-metabolizing enzymes essential for the metabolism of endogenous substrates and xenobiotics. Molecular characteristics of UGTs have been extensively investigated in humans, but in cynomolgus macaques, a non-human primate species widely used in drug metabolism studies, remain to be investigated. In this study, 12 UGT1A cDNAs (UGT1A1, 1A2, 1A4A, 1A4B, 1A5A, 1A5B, 1A5C, 1A6, 1A7, 1A8, 1A9, and 1A10) were isolated and characterized in cynomolgus macaques. UGT1A5C cDNA did not contain a complete coding region due to nonsense mutations, and was excluded from further analysis. Amino acid sequences of all 11 cynomolgus UGT1As had high sequence identities (92-95%) with human UGT1As and were phylogenetically close to human UGT1As. These cynomolgus UGT1A genes shared exons 2-5, and contained a variable exon 1 unique to each gene, similar to human UGT1A genes. Moreover, cynomolgus and human UGT1A gene clusters were located in corresponding regions in the genome. Among the 10 tissue types analyzed, cynomolgus UGT1A mRNAs were most abundantly expressed in the liver, jejunum, and/or kidney, the drug-metabolizing organs, similar to human UGT1As. Among these 11 cynomolgus UGT1A mRNAs, cynomolgus UGT1A2, UGT1A9, and UGT1A10 mRNAs were most abundantly expressed in the liver, kidney, and jejunum, respectively. Cynomolgus liver microsomes and UGT1A proteins catalyzed glucuronidation of the substrates human UGT1As catalyze, including 4-methylumbelliferone, 4-nitrophenol, estradiol, trifluoperazine, serotonin, and propofol, although trifluoperazine glucuronidation was not catalyzed by any cynomolgus UGT1A proteins. These results suggest that cynomolgus UGT1As are functional enzymes with molecular similarities to human UGT1As.

    DOI: 10.1016/j.bcp.2018.06.027

    PubMed

  • Nakanishi Kazuyuki, Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Progesterone hydroxylation by cytochromes P450 2C and 3A enzymes in marmoset liver microsomes. .  Xenobiotica48 ( 8 ) 757 - 763   2018年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Common marmosets (Callithrix jacchus) are potentially useful nonhuman primate models for preclinical drug metabolism studies. However, the roles of marmoset cytochrome P450 (P450) isoforms in the oxidation of endobiotic progesterone have not been fully investigated. In this study, the roles of marmoset P450 isoforms in progesterone hydroxylation were extensively determined. 2. The activities of liver microsomes from individual marmosets with respect to progesterone 21/17alpha- and 16alpha/6beta-hydroxylation were significantly correlated with those for flurbiprofen 4-hydroxylation and midazolam 1'-hydroxylation, respectively, as similar correlations have been found in humans. Anti-P450 2 C and 3 A antibodies suppressed progesterone 21/17alpha- and 16alpha/6beta-hydroxylation, respectively, in marmoset liver microsomes. 3. Recombinant marmoset P450 2C58 and 2C19 catalyzed progesterone to form 21-hydroxyprogesterone and 16alpha-hydroxyprogesterone, respectively, as major products with high maximum velocity/Km values of 0.53 and 0.089 mL/min/nmol, respectively. Recombinant marmoset P450 3A4/90 oxidized progesterone to form 6beta-hydroxyprogesterone as a major product with homotropic cooperativity (>1 of Hill coefficients). 4. These results indicate that the overall activities and roles of liver microsomal P450 enzymes in marmoset livers are similar to those in humans, especially for progesterone 21/17alpha- and 16alpha/6beta-hydroxylation by marmoset P450 2 C and 3 A enzymes, respectively, suggesting important roles for these P450 enzymes in the metabolism of endobiotics in marmosets.

    DOI: 10.1080/00498254.2017.1363444

    PubMed

  • Uno Yasuhiro, Yamazaki Hiroshi .  Expression of cytochrome P450 regulators in cynomolgus macaque. .  Xenobiotica48 ( 7 ) 695 - 703   2018年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Cytochrome P450 (P450) regulators including nuclear receptors and transcription factors have not been fully investigated in cynomolgus macaques, an important species used in drug metabolism studies. In this study, we analyzed 17 P450 regulators by sequence and phylogenetic analysis, and tissue expression. 2. Gene and genome structures of 17 P450 regulators were similar to the human orthologs, and the deduced amino acid sequences showed high sequence identities (92-95%) and more closely clustered in a phylogenetic tree, with the human orthologs. 3. Many of the P450 regulator mRNAs were preferentially expressed in the liver, kidney, and/or jejunum. Among the P450 regulator mRNAs, PXR was most abundant in the liver and jejunum, and HNF4alpha in the kidney. In the liver, the expression of most P450 regulator mRNAs did not show significant differential expression (>2.5-fold) between cynomolgus macaques bred in Cambodia, China, and Indonesia, or rhesus macaques. 4. By correlation analysis, most of the P450 regulators were significantly (p < 0.05) correlated to other P450 regulators, and many of them were also significantly (p < 0.05) correlated with P450s. 5. These results suggest that 17 P450 regulators of cynomolgus macaques had similar molecular characteristics to the human orthologs.

    DOI: 10.1080/00498254.2017.1363928

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Oshio Toru, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Marmoset pulmonary cytochrome P450 2F1 oxidizes biphenyl and 7-ethoxycoumarin and hepatic human P450 substrates. .  Xenobiotica48 ( 7 ) 656 - 662   2018年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. A potentially useful animal model for preclinical studies is the common marmoset (Callithrix jacchus). In this study, using reverse- transcription polymerase chain reaction from marmoset livers, we identified a novel cytochrome P450 (P450) 2F1 cDNA with an open reading frame of 1473 bp. 2. High sequence identities of 92-94% with primate P450 2 F amino acid sequences were indicated by deduced amino acid sequences of P450 2F1 cDNA. Phylogenetic analysis indicates that marmoset P450 2F1 is more congruent with primate P450 2 F forms than those of other species such as rodents. 3. Among five tissue types examined, abundant expression of marmoset P450 2F1 mRNA and P450 2F1 protein in lungs was shown. Cynomolgus monkey P450 2F1 mRNA was abundantly expressed in lungs as well as testes and ovaries in 10 tissue types. 4. Similar to those of humans and cynomolgus monkeys, marmoset P450 2F1 heterologously expressed in Escherichia coli membranes efficiently catalyzed 7-ethoxycoumarin O-deethylation and biphenyl hydroxylation, however unlike human P450 2F1, marmoset P450 2F1 exhibited hydroxylation activity toward coumarin and chlorzoxazone. 5. These findings indicated that P450 2F1 enzyme expressed in marmoset lungs and also catalyzed metabolism of xenobiotics, suggesting the importance of P450 2 F-dependent drug metabolism in marmoset lungs.

    DOI: 10.1080/00498254.2017.1354138

    PubMed

  • Toda Akiko, Uehara Shotaro, Inoue Takashi, Utoh Masahiro, Kusama Takashi, Shimizu Makiko, Uno Yasuhiro, Mogi Masayuki, Sasaki Erika, Yamazaki Hiroshi .  Effects of aging and rifampicin pretreatment on the pharmacokinetics of human cytochrome P450 probes caffeine, warfarin, omeprazole, metoprolol and midazolam in common marmosets genotyped for cytochrome P450 2C19. .  Xenobiotica48 ( 7 ) 720 - 726   2018年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. The pharmacokinetics were investigated for human cytochrome P450 probes after single intravenous and oral administrations of 0.20 and 1.0 mg/kg, respectively, of caffeine, warfarin, omeprazole, metoprolol and midazolam to aged (10-14 years old, n = 4) or rifampicin-treated/young (3 years old, n = 3) male common marmosets all genotyped as heterozygous for a cytochrome P450 2C19 variant. 2. Slopes of the plasma concentration- time curves after intravenous administration of warfarin and midazolam were slightly, but significantly (two-way analysis of variance), decreased in aged marmosets compared with young marmosets. The mean hepatic clearances determined by in silico fitting for individual pharmacokinetic models of warfarin and midazolam in the aged group were, respectively, 23% and 56% smaller than those for the young group. 3. Significantly enhanced plasma clearances of caffeine, warfarin, omeprazole and midazolam were evident in young marmosets pretreated with rifampicin (25 mg/kg daily for 4 days). Two- to three-fold increases in hepatic intrinsic clearance values were observed in the individual pharmacokinetic models. 4. The in vivo dispositions of multiple simultaneously administered drugs in old, young and P450-enzyme-induced marmosets were elucidated. The results suggest that common marmosets could be experimental models for aged, induced or polymorphic P450 enzymes in P450-dependent drug metabolism studies.

    DOI: 10.1080/00498254.2017.1353716

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Yamazaki Hiroshi .  Genetic polymorphisms of drug-metabolizing cytochrome P450 enzymes in cynomolgus and rhesus monkeys and common marmosets in preclinical studies for humans. .  Biochem Pharmacol153   184 - 195   2018年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys (Macaca fascicularis, Old World Monkeys) and common marmosets (Callithrix jacchus, New World Monkeys) have been widely, and expectedly, used as non-human primate models in drug development studies. Major drug-metabolizing cytochrome P450 (P450) enzymes information is now available that supports these primate species as animal models, and it is established that multiple forms of cynomolgus monkey and common marmoset P450 enzymes have generally similar substrate recognition functionality to human P450 enzymes. This research update provides information on genetic polymorphisms of P450 enzymes in cynomolgus monkey and common marmoset like human P450 enzymes. Information on rhesus monkeys (Macaca mulatta), another macaque species used in drug metabolism studies, is also included for comparison. Among a variety of cynomolgus monkey P450 variants investigated, typical examples include individual pharmacokinetic data for efavirenz and R-warfarin associated with cynomolgus monkey P450 2C9 (formerly 2C43) and 2C19 (2C75) variants, respectively, and for R-omeprazole and S-warfarin associated with marmoset P450 2C19 variants. These findings provide a foundation for understanding the individual pharmacokinetic and toxicological results in non-human primates as preclinical models and will help to further support understanding of molecular mechanisms of human P450 function. In addition to these polymorphic P450 enzymes, effects of aging on some drug clearances mediated by cynomolgus monkey and common marmoset P450 enzymes were found in elder animals or animals pretreated with rifampicin. This review describes genetic and acquired individual differences in cynomolgus monkey and common marmoset P450 enzymes involved in drug oxidation associated with pharmacological and/or toxicological effects.

    DOI: 10.1016/j.bcp.2017.12.015

    PubMed

  • Nakanishi Kazuyuki, Uehara Shotaro, Kusama Takashi, Inoue Takashi, Shimura Kanami, Kamiya Yusuke, Murayama Norie, Shimizu Makiko, Uno Yasuhiro, Sasaki Erika, Yamazaki Hiroshi .  In vivo and in vitro diclofenac 5-hydroxylation mediated primarily by cytochrome P450 3A enzymes in common marmoset livers genotyped for P450 2C19 variants. .  Biochem Pharmacol152   272 - 278   2018年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Common marmosets (Callithrix jacchus) are potentially useful nonhuman primate models for preclinical studies. An anti-inflammatory drug, diclofenac is reportedly metabolized mainly by human cytochrome P450 (P450) 2C9 to 4'-hydroxydiclofenac and minorly by P450 3A4 to 5-hydroxydiclofenac that leads to reactive intermediates. In this study, in vivo pharmacokinetics in six marmosets and in vitro metabolism in nine marmoset liver microsomes were analyzed using diclofenac to evaluate marmosets as preclinical drug metabolism models. In wild-type marmosets genotyped for P450 2C19 p.[(Phe7Leu; Ser254Leu; Ile469Thr)], plasma levels of 5-hydroxydiclofenac and 4'-hydroxydiclofenac were roughly similar, but the homozygotes showed approximately three-times higher plasma levels of 5-hydroxydiclofenac than those of 4'-hydroxydiclofenac after oral administrations of diclofenac (50mg/kg). Nine marmoset liver microsomes catalyzed mainly diclofenac 5-hydroxylation with no significant effects of the the P450 2C19 genotype, and partly diclofenac 4'-hydroxylation that depended on the P450 2C19 genotype, at substrate concentrations of 10microM and 100microM. Both Michaels-Menten constant Km values for diclofenac 4'-hydroxylation in human and marmoset liver microsomes were approximately 30muM and those for diclofenac 5-hydroxylation were approximately 120muM. Ketoconazole significantly suppressed only diclofenac 5-hydroxylation in marmoset or human liver microsomes at 0.030muM, indicating main contribution of P450 3A enzymes, which were found to be P450 3A5/90 using recombinant marmoset P450 3A enzymes. These results suggest that marmosets would be a functional animal model for in vivo and in vitro metabolites likely generated via arene oxide intermediates of diclofenac by P450 3A enzymes in humans, primarily because marmosets lack the ortholog of human P450 2C9.

    DOI: 10.1016/j.bcp.2018.04.002

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Mahadhi Hassan M D, Ohura Kayoko, Hosokawa Masakiyo, Imai Teruko .  Molecular characterization and polymorphisms of butyrylcholinesterase in cynomolgus macaques. .  J Med Primatol47 ( 3 ) 185 - 191   2018年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Butyrylcholinesterase (BChE), an enzyme essential for drug metabolism, has been investigated as antidotes against organophosphorus nerve agents, and the efficacy and safety have been studied in cynomolgus macaques. BChE polymorphisms partly account for variable BChE activities among individuals in humans, but have not been investigated in cynomolgus macaques. METHODS: Molecular characterization was carried out by analyzing primary sequence, gene, tissue expression, and genetic variants. RESULTS: In cynomolgus and human BChE, phylogenetically closely related, amino acid residues important for enzyme function were conserved, and gene and genomic structure were similar. Cynomolgus BChE mRNA was most abundantly expressed in liver among the 10 tissue types analyzed. Re-sequencing found 26 non-synonymous genetic variants in 121 cynomolgus and 23 rhesus macaques, indicating that macaque BChE is polymorphic, although none of these variants corresponded to the null or defective alleles of human BChE. CONCLUSIONS: These results suggest molecular similarities of cynomolgus and human BChE.

    DOI: 10.1111/jmp.12342

    PubMed

  • Uehara Shotaro, Yuki Yukako, Uno Yasuhiro, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Terfenadine t-butyl hydroxylation catalyzed by human and marmoset cytochrome P450 3A and 4F enzymes in livers and small intestines. .  Xenobiotica48 ( 4 ) 342 - 347   2018年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Roles of human cytochrome P450 (P450) 3A4 in oxidation of an antihistaminic drug terfenadine have been previously investigated in association with terfenadine-ketoconazole interaction. Several antihistamine drugs have been recently identified as substrates for multiple P450 enzymes. In this study, overall roles of P450 3A4, 2J2, and 4F12 enzymes in terfenadine t-butyl hydroxylation were investigated in small intestines and livers from humans, marmosets, and/or cynomolgus monkeys. 2. Human liver microsomes and liver and small intestine microsomes from marmosets and cynomolgus monkeys effectively mediated terfenadine t-butyl hydroxylation. Ketoconazole and N-hydroxy-N'-(4-butyl-2-methylphenyl)-formamidine (a P450 4A/F inhibitor) almost completely and moderately inhibited these activities, respectively, in human liver microsomes; however, these chemicals did not show substantially suppression in marmoset liver. Anti-human P450 3A and 4F antibodies showed the roughly supportive inhibitory effects. 3. Recombinant P450 3A4/90 and 4F12 showed high terfenadine t-butyl hydroxylation activities with substrate inhibition constants of 84-144 muM (under 26-76 muM of Km values), in similar manners to liver and intestine microsomes. 4. These results suggest that human and marmoset P450 3A4/90 and 4F12 in livers or small intestines played important roles in terfenadine t-butyl hydroxylation. Marmosets could be a model for humans during first pass extraction of terfenadine and related substrates.

    DOI: 10.1080/00498254.2017.1321811

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Yamazaki Hiroshi .  Polymorphisms of cytochrome P450 2B6 (CYP2B6) in cynomolgus and rhesus macaques. .  J Med Primatol   2018年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Cytochrome P450 2B6 (CYP2B6) is an important drug- metabolizing enzyme and is expressed in liver. Although human CYP2B6 variants account for variable enzyme properties among individuals and populations, CYP2B6 genetic variants have not been investigated in cynomolgus macaques, widely used in drug metabolism studies. METHODS: CYP2B6 was resequenced in 120 cynomolgus macaques and 23 rhesus macaques by direct sequencing. RESULTS: Twenty-three non-synonymous variants were found, of which 12 and 3 were unique to cynomolgus macaques and rhesus macaques, respectively. By functional characterization using the 14 variant proteins, 8 variants (V114I, R253C, M435I, V459M, L465P, C475S, R487C, and R487H) showed different rate (>1.5-fold) of testosterone 16beta-hydroxylation to wild type. However, the four variants (M435I, L465P, C475S, and R487H) were analyzed in liver microsomes, and the catalytic rates were not substantially different from wild type. CONCLUSIONS: Macaque CYP2B6 was polymorphic, and the genotype could partly account for variable enzyme activities of macaque CYP2B6.

    DOI: 10.1111/jmp.12336

    PubMed

  • Utoh Masahiro, Kusama Takashi, Miura Tomonori, Mitsui Marina, Kawano Mirai, Hirano Takahiro, Shimizu Makiko, Uno Yasuhiro, Yamazaki Hiroshi .  R-warfarin clearances from plasma associated with polymorphic cytochrome P450 2C19 and simulated by individual physiologically based pharmacokinetic models for 11 cynomolgus monkeys. .  Xenobiotica48 ( 2 ) 206 - 210   2018年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Cynomolgus monkey cytochrome P450 2C19 (formerly known as P450 2C75), homologous to human P450 2C19, has been identified as R-warfarin 7-hydroxylase. In this study, simulations of R-warfarin clearance in individual cynomolgus monkeys genotyped for P450 2C19 p.[(Phe100Asn; Ala103Val; Ile112Leu)] were performed using individual simplified physiologically based pharmacokinetic (PBPK) modeling. 2. Pharmacokinetic parameters and absorption rate constants, volumes of the systemic circulation, and hepatic intrinsic clearances for individual PBPK models were estimated for eleven cynomolgus monkeys. 3. One-way ANOVA revealed significant effects of the genotype (p < 0.01) on the observed elimination half-lives and areas under the curves of R-warfarin among the homozygous mutant, heterozygous mutant, and wild-type groups. R-Warfarin clearances in individual cynomolgus monkeys genotyped for P450 2C19 were simulated by simplified PBPK modeling. The modeled hepatic intrinsic clearances were significantly associated with the P450 2C19 genotypes. The liver microsomal elimination rates of R-warfarin for individual animals after in vivo administration showed significant reductions associated with the genotype (p < 0.01). 4. This study provides important information to help simulate clearances of R-warfarin and related medicines associated with polymorphic P450 2C19 in individual cynomolgus monkeys, thereby facilitating calculation of the fraction of hepatic clearance.

    DOI: 10.1080/00498254.2017.1288945

    PubMed

  • Murayama Norie, Yajima Kanako, Hikawa Mikiko, Shimura Kanami, Ishii Yu, Takada Masaki, Uno Yasuhiro, Utoh Masahiro, Iwasaki Kazuhide, Yamazaki Hiroshi .  Assessment of multiple cytochrome P450 activities in metabolically inactivated human liver microsomes and roles of P450 2C isoforms in reaction phenotyping studies. .  Biopharm Drug Dispos39 ( 2 ) 116 - 121   2018年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The fraction of substrate metabolized (fm ) can be used to estimate drug interactions and can be determined by comparison of the intrinsic clearances (CLint ) of victim drugs obtained from inhibited and uninhibited hepatic enzymes. Commercially available human liver microsomes were recently developed in which one cytochrome P450 (P450) isoform is selectively inactivated. These inactivated liver microsomes were used to evaluate the roles of P450 2C isoforms in the depletion and oxidation of probe substrates. Determination of CLint with sets of control and P450 2C9-inactivated liver microsomes yielded fm,P450 2C9 values of 0.69-1.0 for celecoxib, diclofenac and warfarin. Apparent minor contributions of P450 1A2/2C8/3A4 were seen in depletion assays, yielding ~1 for the sum of the fm values. Selectively inactivated liver microsomes were thereby shown to be potentially useful for determining the in vitro fm values for major P450 2C9 contributions to substrate oxidations. Metabolite formations from diclofenac and warfarin were suppressed by 62-84% by the replacement of control liver microsomes with P450 2C9-inactivated liver microsomes. R-, S- and racemic omeprazole and troglitazone oxidation activities by liver microsomes at multiple substrate concentrations were suppressed by 26-36% and 22-50%, respectively, when P450 2C19- and 2C8-inactivated liver microsomes were used in place of control liver microsomes. This study provides important information to help elucidate the different roles of P450 isoforms in metabolite formation at different substrate concentrations. The data obtained allow the fractions metabolized to be calculated for victim drugs.

    DOI: 10.1002/bdd.2115

    PubMed

  • Uno Y, Osada N, Sakurai S, Shimozawa N, Iwata T, Ikeo K, Yamazaki H .  Development of genotyping method for functionally relevant variants of cytochromes P450 in cynomolgus macaques. .  J Vet Pharmacol Ther41 ( 1 ) e30 - e34   2018年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In cynomolgus macaques (Macaca fascicularis), widely used in drug metabolism studies, CYP2C9, CYP2C76, CYP2D6, CYP3A4, and CYP3A5, important drug-metabolizing enzymes, are abundantly expressed in liver and metabolize cytochrome P450 substrates. CYP2C9 (c.334A>C), CYP2C76 (c.449TG>A), CYP2D6 (c.891A>G), CYP3A4 (IVS3 + 1G>del), and CYP3A5 (c.625A>T) substantially influence metabolic activity of enzymes, and thus are important variants in drug metabolism studies. In this study, a real-time PCR method was developed for genotyping these variants. The validity of the methods was verified by genotyping two wild type, two heterozygous, and two homozygous DNAs and was used to genotype 41 cynomolgus macaques (from Cambodia, Indonesia, the Philippines, or Vietnam) for the five variants, along with another important variant CYP2C19 (c.308C>T). The CYP2C9 and CYP2C19 variants were found only in Cambodian and Vietnamese animals, while the CYP2C76 and CYP2D6 variants were found only in Indonesian and Philippine animals. The CYP3A4 and CYP3A5 variants were not found in any of the animals analyzed. Mauritian animals, genotyped using next-generation sequencing data for comparison, possessed the CYP2C19 and CYP2D6 variants, but not the other variants. These results indicated differences in prevalence of these important variants among animal groups. Therefore, the genotyping tool developed is useful for drug metabolism studies using cynomolgus macaques.

    DOI: 10.1111/jvp.12443

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Molecular cloning and tissue distribution of a novel marmoset ABC transporter. .  Biopharm Drug Dispos39 ( 1 ) 59 - 63   2018年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Common marmosets (Callithrix jacchus) have been recognized as a useful small non-human primate model in preclinical testing for drug development. In this study, a cDNA of novel ATP-dependent efflux transporter ABCB1 was cloned from marmoset liver tissue. Marmoset ABCB1 cDNA encodes a protein of 1279 amino acid residues (MW = 141.4 kDa) containing characteristic regions of an ATP-binding cassette (ABC) protein, two hydrophobic transmembrane regions and two cytoplasmic nucleotide-binding regions, similar to human ABCB1. The deduced amino acid sequences were more highly identical (95%) to those of human ABCB1 compared with non-primate species such as dogs, pigs and rodents (79-90%). A close evolutionary relationship of ABCB1 among marmosets, cynomolgus and rhesus monkeys and humans was evident from a phylogenetic analysis using ABCB1 amino acid sequences from primates, dogs, pigs and rodents. Tissue distribution analyses by quantitative reverse transcription-polymerase chain reaction indicated that marmoset ABCB1 mRNA was most abundant in kidneys, followed by small intestines and livers, similar to human ABCB1, and marmoset ABCB1 proteins in these tissues were also detected by immunoblotting. These results indicated that the primary structure and tissue distribution of ABCB1 in marmosets were similar to those of humans, suggesting similar molecular characteristics of ABCB1 between marmosets and humans.

    DOI: 10.1002/bdd.2111

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Yamazaki Hiroshi .  Hepatic expression of cytochrome P450 enzymes in non-human primate species. .  J Med Primatol46 ( 6 ) 347 - 351   2017年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cytochromes P450 (P450) largely remain to be characterized in great apes. Comparative immunochemical detection of drug metabolizing forms of P450s 1A, 2A, 2B, 2C, 2D, 2E, 2J, 3A, 4A, and 4F in liver microsomes from chimpanzees, gorillas, orangutans, gibbons, cynomolgus and rhesus macaques, and common marmosets were carried out.

    DOI: 10.1111/jmp.12288

    PubMed

  • Okamatsu Gaku, Kawakami Kei, Komatsu Tetsuya, Kitazawa Takio, Uno Yasuhiro, Teraoka Hiroki .  Functional expression and comparative characterization of four feline P450 cytochromes using fluorescent substrates. .  Xenobiotica47 ( 11 ) 951 - 961   2017年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Cytochrome P450s (CYP) are a major group of metabolizing enzymes for xenobiotics in humans and other mammals. The properties of CYP isoforms in the domestic cat, an obligate carnivore, are largely unknown at present. In this study, we studied relative expression in tissues and enzymatic properties of nine significant feline CYP isoforms. 2. CYP2E2 transcript was most abundant in the feline liver, followed by CYP2A13 and 2E1. Transcripts of CYP3A131, 1A2 and 1A1 were also present in the liver, while CYP2D6 and 3A132 were only slightly expressed. CYP3A131 was a major transcript in the small intestine. 3. Four major CYP isoforms in the feline liver and small intestine (CYP1A2, CYP2A13, CYP2E2 and CYP3A131) were heterologously expressed in Escherichia coli to generate functional monooxygenase systems. We carried out screenings of 17 test compounds known to be inhibitors of CYP isoforms in other mammals as well as two anticancer drugs to assess the activity modulation of feline CYP isoforms using fluorogenic substrates. These CYP isoforms showed similar selectivity to counterparts in other mammals against inhibitors as a whole but with many exceptions. 4. The present study suggests the usefulness of the feline CYP recombinant system to obtain chemical affinity information and possible drug interactions in CYP metabolism of domestic cats.

    DOI: 10.1080/00498254.2016.1257172

    PubMed

  • Utoh Masahiro, Miura Tomonori, Kusama Takashi, Uehara Shotaro, Shimizu Makiko, Uno Yasuhiro, Yamazaki Hiroshi .  Efavirenz clearances in vitro and in vivo in six cynomolgus monkeys associated with polymorphic cytochrome P450 2C9 and simulated by individual physiologically based pharmacokinetic models. .  Biopharm Drug Dispos38 ( 7 ) 439 - 442   2017年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkey cytochrome P450 2C9 (formerly known as P450 2C43) variation was reportedly associated with metabolic clearance of the antiretroviral drug efavirenz in vivo (in three wild-type, one heterozygote and two homozygote animals), being unlikely in the case of human P450 2B6-dependent efavirenz clearance. In this study, the liver microsomal elimination rates of efavirenz for the same individual animals previously treated with intravenous/oral administrations of efavirenz showed significant reductions associated with the P450 2C9 p.[(I112L)] genotype (p < 0.05). Simulations of efavirenz clearance after oral administrations in individual cynomolgus monkeys were performed using individual simplified physiologically based pharmacokinetic (PBPK) modeling consisting of gut, liver and central compartments. The modeled hepatic intrinsic clearances were also significantly associated with the P450 2C9 genotypes, however, absorption rate constants or volumes of the systemic circulation were not likely determining factors for the individual efavirenz clearance variations in the six cynomolgus monkeys. This study provides important information to help simulate the clearances of efavirenz and related medicines associated with polymorphic P450 2C9 in individual cynomolgus monkeys, thereby facilitating the calculation of the fraction of liver microsomal clearance for estimating in vivo drug clearance with simplified PBPK modeling.

    DOI: 10.1002/bdd.2084

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Tomioka Etsuko, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Functional characterization and tissue expression of marmoset cytochrome P450 2E1. .  Biopharm Drug Dispos38 ( 6 ) 394 - 397   2017年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Common marmosets (Callithrix jacchus) have attracted increasing attention as a useful small non-human primate model in preclinical research. However, studies on marmoset cytochrome P450 (P450) 2E enzyme have scarcely been conducted. In this study, the full-length cDNA encoding P450 2E1 enzyme was isolated from marmoset livers by reverse transcription (RT)-polymerase chain reaction (PCR). Marmoset P450 2E1 amino acid sequences were highly identical (>88%) to those of cynomolgus monkey and human P450 2E1 enzymes. Phylogenetic analysis indicated a close evolutionary relationship among marmoset, cynomolgus monkey, and human P450 2E1 enzymes. The tissue expression pattern analyzed by real- time RT-PCR and immunoblotting demonstrated that marmoset P450 2E1 mRNA and proteins were predominantly expressed in livers. Marmoset P450 2E1 enzyme heterologously expressed in Escherichia coli catalyzed the hydroxylation of p-nitrophenol, chlorzoxazone, and theophylline, similar to cynomolgus monkey and human P450 2E1 enzymes. By kinetic analyses, those P450 2E1 enzymes catalyzed p-nitrophenol hydroxylation with similar affinities and relatively high intrinsic clearance efficiencies. These results indicated that tissue distribution and enzyme-substrate specificity of marmoset P450 2E1 were similar to cynomolgus monkey and human P450 2E1 enzymes, suggesting that marmosets are a suitable primate model for P450 2E1-dependent drug and xenobiotic metabolism.

    DOI: 10.1002/bdd.2080

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Okamoto Eriko, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Molecular Cloning and Characterization of Marmoset Aldehyde Oxidase. .  Drug Metab Dispos45 ( 8 ) 883 - 886   2017年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Common marmosets (Callithrix jacchus), New World monkeys, are a promising primate model for preclinical drug metabolism studies because of the similarities of cytochrome P450 (P450) enzyme function to those of humans. Despite an increasing number of drug candidates catalyzed by non-P450 enzymes, drug metabolizing enzymes other than P450s have been hardly identified or characterized in marmosets. In this study, we identified aldehyde oxidase (AOX) 1 gene by marmoset genome analysis. AOX1 cDNA was cloned from marmoset livers by reverse transcription- polymerase chain reaction. Deduced amino acid sequences of AOX1 cDNA showed high sequence identities (92-93%) with primate AOX1s. Phylogenetic analysis showed that marmoset AOX1 was closely clustered with primate AOX1s, unlike nonprimate animal model AOX1s of pig, rabbit, rat, and mouse used in drug metabolism. The tissue expression analyses by real- time reverse-transcription polymerase chain reaction and immunoblotting showed that marmoset AOX1 mRNA and protein were abundantly expressed in livers, similar to cynomolgus monkeys and humans. Marmoset AOX1 heterologously expressed in Escherichia coli catalyzed the oxidation of carbazeran and phthalazine, typical AOX1 substrates, similar to cynomolgus monkey and human AOX1s. Human and marmoset AOX1 effectively catalyzed phthalazine oxidation when assessed with Michaelis-Menten kinetics, but cynomolgus monkey AOX1 catalyzed this reaction with cooperative kinetics with high capacity. These results indicated that tissue distribution and enzymatic function of AOX1 enzyme is similar between marmosets and humans, suggesting that marmosets are a suitable primate model for AOX-dependent drug metabolism in preclinical studies.

    DOI: 10.1124/dmd.117.076042

    PubMed

  • Uehara Shotaro, Ishii Sakura, Uno Yasuhiro, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Regio- and Stereo-Selective Oxidation of a Cardiovascular Drug, Metoprolol, Mediated by Cytochrome P450 2D and 3A Enzymes in Marmoset Livers. .  Drug Metab Dispos45 ( 8 ) 896 - 899   2017年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A beta-blocker, metoprolol, is one of the in vivo probes for human cytochrome P450 (P450) 2D6. Investigation of nonhuman primate P450 enzymes helps to improve the accuracy of the extrapolation of pharmacokinetic data from animals into humans. Common marmosets (Callithrix jacchus) are a potential primate model for preclinical research, but the detailed roles of marmoset P450 enzymes in metoprolol oxidation remain unknown. In this study, regio- and stereo-selectivity of metoprolol oxidations by a variety of P450 enzymes in marmoset and human livers were investigated in vitro. Although liver microsomes from cynomolgus monkeys and rats preferentially mediated S-metoprolol O-demethylation and R-metoprolol alpha-hydroxylation, respectively, those from humans, marmosets, minipigs, and dogs preferentially mediated R-metoprolol O-demethylation, in contrast to the slow rates of R- and S-metoprolol oxidation in mouse liver microsomes. R- and S-metoprolol O-demethylation activities in marmoset livers were strongly inhibited by quinidine and ketoconazole, and were significantly correlated with bufuralol 1'-hydroxylation and midazolam 1'-hydroxylation activities and also with P450 2D and 3A4 contents, which is different from the case in human livers that did not have any correlations with P450 3A-mediated midazolam 1'-hydroxylation. Recombinant human P450 2D6 enzyme and marmoset P450 2D6/3A4 enzymes effectively catalyzed R-metoprolol O-demethylation, comparable to the activities of human and marmoset liver microsomes, respectively. These results indicated that the major roles of P450 2D enzymes for the regio- and stereo-selectivity of metoprolol oxidation were similar between human and marmoset livers, but the minor roles of P450 3A enzymes were unique to marmosets.

    DOI: 10.1124/dmd.117.075630

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Ishii Sakura, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Marmoset cytochrome P450 4A11, a novel arachidonic acid and lauric acid omega-hydroxylase expressed in liver and kidney tissues. .  Xenobiotica47 ( 7 ) 553 - 561   2017年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. A cDNA encoding novel cytochrome P450 (P450) 4A enzyme was cloned from marmoset livers by reverse transcription (RT)-polymerase chain reaction (PCR) based on the marmoset genome sequences. The amino acid sequence deduced from P450 4A11 cDNA contained consensus sequences of six substrate recognition sites and one heme-binding domain. 2. Marmoset P450 4A11, highly identical (85-88%) to cynomolgus monkey and human P450 4A enzymes, was grouped into the same cluster as cynomolgus monkey and human P450 4A enzymes by phylogenetic analysis. 3. The tissue distribution analyses by real-time RT PCR and immunoblotting demonstrated that marmoset P450 4A11 mRNA and proteins were expressed in kidneys and livers. Marmoset P450 4A11 enzyme heterologously expressed in Escherichia coli preferentially catalyzed the omega-hydroxylation of arachidonic acid and lauric acid, similar to cynomolgus monkey and human P450 4A11 enzymes. However, lauric acid omega-hydroxylation activity of marmoset P450 4A11 was low compared with those of marmoset liver microsomes. 4. These results indicated that novel marmoset P450 4A11 was also a fatty acid omega-hydroxylase expressed in kidneys and livers, with the same regioselectivity (at omega-position of fatty acid) as cynomolgus monkey and human P450 4A enzymes.

    DOI: 10.1080/00498254.2016.1206673

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Nakanishi Kazuyuki, Ishii Sakura, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Marmoset Cytochrome P450 3A4 Ortholog Expressed in Liver and Small- Intestine Tissues Efficiently Metabolizes Midazolam, Alprazolam, Nifedipine, and Testosterone. .  Drug Metab Dispos45 ( 5 ) 457 - 467   2017年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Common marmosets (Callithrix jacchus), small New World primates, are increasingly attracting attention as potentially useful animal models for drug development. However, characterization of cytochrome P450 (P450) 3A enzymes involved in the metabolism of a wide variety of drugs has not investigated in marmosets. In this study, sequence homology, tissue distribution, and enzymatic properties of marmoset P450 3A4 ortholog, 3A5 ortholog, and 3A90 were investigated. Marmoset P450 3A forms exhibited high amino acid sequence identities (88-90%) to the human and cynomolgus monkey P450 3A orthologs and evolutionary closeness to human and cynomolgus monkey P450 3A orthologs compared with other P450 3A enzymes. Among the five marmoset tissues examined, P450 3A4 ortholog mRNA was abundant in livers and small intestines where P450 3A4 ortholog proteins were immunologically detected. Three marmoset P450 3A proteins heterologously expressed in Escherichia coli membranes catalyzed midazolam 1'- and 4-hydroxylation, alprazolam 4-hydroxylation, nifedipine oxidation, and testosterone 6beta-hydroxylation, similar to cynomolgus monkey and human P450 3A enzymes. Among the marmoset P450 3A enzymes, P450 3A4 ortholog effectively catalyzed midazolam 1'-hydroxylation, comparable to microsomes from marmoset livers and small intestines. Correlation analyses with 23 individual marmoset liver microsomes suggested contributions of P450 3A enzymes to 1'-hydroxylation of both midazolam (human P450 3A probe) and bufuralol (human P450 2D6 probe), similar to cynomolgus monkey P450 3A enzymes. These results indicated that marmoset P450 3A forms had functional characteristics roughly similar to cynomolgus monkeys and humans in terms of tissue expression patterns and catalytic activities, suggesting marmosets as suitable animal models for P450 3A-dependent drug metabolism.

    DOI: 10.1124/dmd.116.074898

    PubMed

  • Uehara Shotaro, Shimizu Makiko, Uno Yasuhiro, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Marmoset Flavin-Containing Monooxygenase 3 in the Liver Is a Major Benzydamine and Sulindac Sulfide Oxygenase. .  Drug Metab Dispos45 ( 5 ) 497 - 500   2017年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Common marmosets (Callithrix jacchus) are potentially primate models for preclinical drug metabolism studies because there are similarities in the molecular characteristics of cytochrome P450 enzymes between this species and humans. However, characterization of non-cytochrome P450 enzymes has not been clarified in marmosets. Here, we report characterization of flavin-containing monooxygenases FMO1-FMO5 identified in marmoset tissues. Marmoset FMO forms shared high amino acid sequence identities (93%-95%) and phylogenetic closeness with human homologous FMO forms. FMO1 and FMO3 mRNA were abundantly expressed in the liver and kidneys among five marmoset tissues examined, where FMO3 protein was detected by immunoblotting. FMO inhibition assays using preheated tissue microsomes indicated that benzydamine N-oxygenation and sulindac sulfide S-oxygenation in the marmoset liver was mainly catalyzed by FMO3, the major hepatic FMO. Marmoset FMO3 protein heterologously expressed in Escherichia coli effectively catalyzed benzydamine N-oxygenation and sulindac sulfide S-oxygenation comparable to marmoset liver microsomes. These results indicate that the FMO3 enzyme expressed in marmoset livers mainly metabolizes benzydamine and sulindac sulfide (typical human FMO substrates), suggesting its importance for FMO-dependent drug metabolism in marmosets.

    DOI: 10.1124/dmd.117.075184

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Cloning and expression of a novel catechol-O-methyltransferase in common marmosets. .  J Vet Med Sci79 ( 2 ) 267 - 272   2017年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Catechol-O-methyltransferase (COMT) catalyzes the O-methylation of endogenous catechol amines and estrogens and exogenous catechol-type of drugs. A Parkinson's disease model of common marmoset (Callithrix jacchus) has been widely used in preclinical studies to evaluate inhibitory potential of new drug candidates on marmoset COMT. Despite COMT inhibitors could potentiate the pharmacological action of levodopa on Parkinson's disease in animal models, marmoset COMT cDNA has not yet been identified and characterized. In this study, a cDNA highly homologous to human COMT was cloned from marmoset livers. This cDNA encoded 268 amino acids containing a transmembrane region and critical amino acid residues for catalytic function. The amino acid sequences of marmoset COMT shared high sequence identity (90%) with human COMT. COMT mRNA was expressed in all five tissues tested, including brain, lung, liver, kidney and small intestine, and was more abundant in marmoset liver and kidney. Membrane-bound COMT was immunochemically detected in livers and kidneys, whereas soluble COMT was detected in livers, similar to humans. These results indicated that the molecular characteristics of marmoset COMT were generally similar to the human ortholog.

    DOI: 10.1292/jvms.16-0459

    PubMed

  • Uno Yasuhiro, Takata Ryo, Kito Go, Yamazaki Hiroshi, Nakagawa Kazuko, Nakamura Yusuke, Kamataki Tetsuya, Katagiri Toyomasa .  Sex- and age-dependent gene expression in human liver: An implication for drug-metabolizing enzymes. .  Drug Metab Pharmacokinet32 ( 1 ) 100 - 107   2017年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Sex and age differences in hepatic expression of drug-metabolizing enzyme genes could cause variations in drug metabolism, but has not been fully elucidated, especially in Asian population. In this study, the global expression of human hepatic genes was analyzed by microarrays in 40 Japanese subjects (27 males and 13 females). Thirty-five sex-biased genes were identified (P < 0.005). Whereas, 60 age-biased genes in two age groups, <60 years and >/=70 years (P < 0.001), were identified in males. By Gene Ontology analysis, the sex-biased genes were related to protein catabolism and modification, while the age-biased genes were related to transcription regulation and cell death. Quantitative polymerase chain reaction confirmed the female-biased expression of drug-metabolizing enzyme genes BChE, CYP4X1, and SULT1E1 (>/=1.5-fold, P < 0.05). Further analysis of drug-metabolizing enzyme genes indicated that expression of CYP2A6 and CYP3A4 in females in the >/=70 age group was less than in the <60 age group (>/=1.5-fold, P < 0.05), and this trend was also observed for PXR expression in males (>/=1.5-fold, P < 0.05). The results presented provide important insights into hepatic physiology and function, especially drug metabolism, with respect to sex and age.

    DOI: 10.1016/j.dmpk.2016.10.409

    PubMed

  • Okamatsu Gaku, Komatsu Tetsuya, Ono Yuka, Inoue Hiroki, Uchide Tsuyoshi, Onaga Takenori, Endoh Daiji, Kitazawa Takio, Hiraga Takeo, Uno Yasuhiro, Teraoka Hiroki .  Characterization of feline cytochrome P450 2B6. .  Xenobiotica47 ( 2 ) 93 - 102   2017年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Little is known about drug metabolism in carnivores. Although the domestic cat (Felis catus) is an obligate carnivore and is the most common companion animal, usage and dosage of many drugs are determined according to information obtained from humans and dogs. We determined the complete cDNA sequence of CYP2B6 from the feline lung. 2. Feline CYP2B6 consists of 494 deduced amino acids, showing highest identity with the dog CYP2B ortholog, followed by those of horse, pig, primate and human. 3. Feline CYP2B6 transcripts were expressed predominantly in the lung and slightly in the small intestine but not in the liver without significant sex-dependent differences. Western blot analysis with an anti-human CYP2B6 antibody confirmed the presence of CYP2B protein in the lung but not in the liver. 4. Feline CYP2B6 proteins heterologously expressed in Escherichia coli metabolized several substrates specific to human CYP2B6, including 7-ethoxy-4-(trifluoromethyl) coumarin (EFC). The metabolic activity was strongly inhibited by medetomidine and atipamezole, potent inhibitors of canine CYP2B11 (now officially CYP2B6) as well as by ticlopidine and sertraline, inhibitors selective to human CYP2B6. 5. The results suggest that feline CYP2B6 is a functional CYP2B ortholog that plays a role in the local defense mechanism in the cat respiratory system and intestine.

    DOI: 10.3109/00498254.2016.1145754

    PubMed

  • Koyama Shuzo, Fukuda Koji, Watanabe Sho, Matsushita Akinori, Tsuchiya Hideaki, Fujinami Nahoko, Kohara Sakae, Murayama Norie, Nagano Masashi, Yamazaki Hiroshi, Fukuzaki Koichiro, Uno Yasuhiro, Hosoi Yoshihiko .  CYP2C76 deficiency is embryonic lethal in cynomolgus macaques: The potential role of CYP2C76 in early embryogenesis. .  Drug Metab Pharmacokinet32 ( 1 ) 112 - 115   2017年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus macaques are an important primate species for drug metabolism studies; however cynomolgus CYP2C76, an important drug-metabolizing enzyme, accounts for drug metabolism differences to humans, so that CYP2C76-null animals might show drug-metabolizing properties more similar to humans. In this study, attempts were made to produce CYP2C76-null animals by assisted reproduction technology. Oocytes and sperm collected from the heterozygotes for the null allele (c.449TG > A) were subjected to intracytoplasmic sperm injection, and the embryos produced were cultured in vitro through the blastocyst stage. Preimplantation genetic diagnosis using a biopsied portion of the blastocyst revealed that none of the 32 blastocysts analyzed were homozygotes. In contrast, 2 of the 20 embryos analyzed were homozygotes at the 8-cell stage, indicating that CYP2C76-null embryos most likely stop developing between the 8-cell and blastocyst stage. By polymerase chain reaction, expression of CYP2C76 mRNA was detected in oocytes and blastocysts, but not in 2-, 4-, 8-, or 16/32-cell stage embryos. Metabolic assays showed that CYP2C76 metabolized progesterone. These results indicated that CYP2C76 null was likely embryonic lethal, suggesting its potential role during early embryogenesis in cynomolgus macaques.

    DOI: 10.1016/j.dmpk.2016.10.411

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Suzuki Takako, Inoue Takashi, Utoh Masahiro, Sasaki Erika, Yamazaki Hiroshi .  Strong Induction of Cytochrome P450 1A/3A, But not P450 2B, in Cultured Hepatocytes from Common Marmosets and Cynomolgus Monkeys by Typical Human P450 Inducing Agents. .  Drug Metab Lett10 ( 4 ) 244 - 253   2017年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Common marmosets (Callithrix jacchus) and cynomolgus monkeys (Macaca fascicularis) are used as non-human primate models in preclinical studies for drug development. OBJECTIVE: The assessment of P450 induction in hepatocytes from marmosets and cynomolgus monkeys was performed using typical P450 inducers. METHODS: Induction of cytochrome P450 1-4 family enzymes was analyzed in two lots of cultured hepatocytes from common marmosets and cynomolgus monkeys after 24-h treatment with typical human P450 inducing agents by real-time reverse transcription-polymerase chain reaction. RESULTS: Marmoset P450 3A4 mRNA and P450 2C8/2C19 mRNA in hepatocytes were strongly (>10- fold) and weakly (>2) induced by rifampicin, respectively. Marmoset 1A1 and 1A2 mRNA were induced strongly (>200-fold) by beta-naphthoflavone and omeprazole. Marmoset P450 2B6 mRNA was induced (~5-fold) by a constitutive androstane receptor agonist, but not by phenobarbital. Cynomolgus monkey P450 3A4 mRNA and P450 1A1 mRNA in cultured hepatocytes were also induced by rifampicin and omeprazole, respectively, but P450 2B6 mRNA was not induced by phenobarbital. CONCLUSION: These results indicate that P450 1A/3A induction by typical human P450 inducers in hepatocytes from marmosets and/or cynomolgus monkeys are similar to those of humans (except for P450 2B induction by phenobarbital in humans), suggesting that marmosets and cynomolgus monkeys might be suitable models for evaluating the drug interactions in preclinical studies.

    DOI: 10.2174/1872312810666161114144412,

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Yamazaki Hiroshi .  Utility of non-human primates in drug development: Comparison of non- human primate and human drug-metabolizing cytochrome P450 enzymes. .  Biochem Pharmacol121   1 - 7   2016年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys (Macaca fascicularis, an Old World Monkey) have been widely used as a non-human primate model in preclinical studies because of their genetic and physiological similarity to humans. This trend has been followed by common marmoset (Callithrix jacchus, a New World Monkey). However, drug-metabolism properties in these non-human primates have not been fully understood due to limited information on cytochrome P450 (P450) enzymes, major drug-metabolizing enzymes in humans. Multiple forms of cynomolgus monkey P450 enzymes have been identified and characterized in comparison to those of humans, including a cynomolgus monkey specific form, P450 2C76. Similarly, marmoset P450 1A/B, 2A, 2C, 2D, and 4F enzymes were recently identified and characterized to understand drug metabolism properties. In this research update, updates for marmoset, cynomolgus monkey, and human P450 cDNAs are provided. Marmoset and cynomolgus monkey P450 enzymes showed high sequence homology to their human counterparts and generally had similar substrate recognition functionality to human P450 enzymes; however, they also possibly contribute to limited specific differences in drug oxidative metabolism partly due to small differences in amino acid residues. These findings provide a foundation for successful use of non-human primates as preclinical models and will help to further understand molecular mechanisms of human P450 function. In addition to the P450 enzymes, flavin-containing monooxygenases, another monooxygenase family, in these non-human primates have been found to be involved in the oxidation of a variety of compounds associated with pharmacological and/or toxicological effects in humans and are also described.

    DOI: 10.1016/j.bcp.2016.06.008

    PubMed

  • Utoh Masahiro, Suemizu Hiroshi, Mitsui Marina, Kawano Mirai, Toda Akiko, Uehara Shotaro, Uno Yasuhiro, Shimizu Makiko, Sasaki Erika, Yamazaki Hiroshi .  Human plasma concentrations of cytochrome P450 probe cocktails extrapolated from pharmacokinetics in mice transplanted with human hepatocytes and from pharmacokinetics in common marmosets using physiologically based pharmacokinetic modeling. .  Xenobiotica46 ( 12 ) 1049 - 1055   2016年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. The pharmacokinetic data of cytochrome P450 probes in humans can be extrapolated from corresponding data in cynomolgus monkeys, dogs and minipigs using simplified physiologically based pharmacokinetic (PBPK) modeling. In this study, the modeling methodology was further adapted to estimate human plasma concentrations of P450 probes based on data from mice transplanted with human hepatocytes or based on data from marmosets. 2. Using known species allometric scaling factors, the observed plasma concentrations of caffeine, warfarin, omeprazole, metoprolol, and midazolam in chimeric TK-NOG mice with humanized liver were scaled to human oral monitoring equivalents. Using the same approach, the previously reported pharmacokinetics of the five P450 probes in marmosets were also scaled to reported equivalents in humans using in vitro metabolic clearance data. 3. Human plasma concentration profiles of the five P450 probes estimated by simplified human PBPK models based on the observed pharmacokinetics in mice with humanized liver and on the reported pharmacokinetics in marmosets were consistent with previously published pharmacokinetic data in Caucasians. 4. These results suggest that mice with humanized liver and/or marmosets could be suitable pharmacokinetic models for humans during research into new drugs, especially when used in combination with simple PBPK models.

    DOI: 10.3109/00498254.2016.1147102

    PubMed

  • Uehara Shotaro, Kawano Mirai, Murayama Norie, Uno Yasuhiro, Utoh Masahiro, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Oxidation of R- and S-omeprazole stereoselectively mediated by liver microsomal cytochrome P450 2C19 enzymes from cynomolgus monkeys and common marmosets. .  Biochem Pharmacol120   56 - 62   2016年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Racemic omeprazole has been used for clinically treating gastric acid- related diseases and also as a typical human cytochrome P450 (P450) 2C19 probe substrate in preclinical studies. S-Omeprazole has been developed as a single enantiomer medicine, which has been reported not to be associated with polymorphic human P450 2C19 phenotypes. In this study, 5-hydroxylation and sulfoxidation activities, with respect to stereoselective R- and S-omeprazole oxidations by liver microsomes from experimental animals including non-human primates and humans, were investigated in vitro. Liver microsomes from humans, cynomolgus monkeys, and mice preferentially mediated R-omeprazole 5-hydroxylations, however those from marmosets, minipigs, dogs, and rats preferentially mediated S-omeprazole 5-hydroxylations. High catalytic activities were observed for recombinant human P450 2C19 in R-omeprazole 5-hydroxlations, cynomolgus monkey P450 2C19 in both R- and S-omeprazole 5-hydroxlations, and marmoset P450 2C19 in S-omeprazole 5-hydroxlations. On the other hand, human, cynomolgus monkey, and marmoset P450 3A enzymes preferentially mediated S-omeprazole sulfoxidations. Correlation and kinetic analyses revealed a high affinity of polymorphic cynomolgus monkey and marmoset liver microsomal P450 2C19 enzymes with respect to R- and S-omeprazole 5-hydroxylations, respectively, and a high capacity of cynomolgus monkey and marmoset liver microsomal P450 3A4 for omeprazole 5-hydroxylations and sulfoxidations. R-and S-omeprazole 5-hydroxylation activities in cynomolgus monkey and marmoset liver microsomes were significantly different among wild-type, heterozygous, and homozygous animals genotyped for cynomolgus monkey P450 2C19 p.[(Phe100Asn; Ala103Val; Ile112Leu)] and for marmoset P450 2C19 p.[(Phe7Leu; Ser254Leu; Ile469Thr)], respectively. The results of this study demonstrate polymorphic cynomolgus monkey and marmoset P450 2C19-dependent omeprazole oxidation activities with individual variations similar to humans.

    DOI: 10.1016/j.bcp.2016.09.010

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Okamoto Eriko, Sasaki Erika, Yamazaki Hiroshi .  Marmoset cytochrome P450 2J2 mainly expressed in small intestines and livers effectively metabolizes human P450 2J2 probe substrates, astemizole and terfenadine. .  Xenobiotica46 ( 11 ) 977 - 985   2016年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Common marmoset (Callithrix jacchus), a New World Monkey, has potential to be a useful animal model in preclinical studies. However, drug metabolizing properties have not been fully understood due to insufficient information on cytochrome P450 (P450), major drug metabolizing enzymes. 2. Marmoset P450 2J2 cDNA was isolated from marmoset livers. The deduced amino acid sequence showed a high-sequence identity (91%) with cynomolgus monkey and human P450 2J2 enzymes. A phylogenetic tree revealed that marmoset P450 2J2 was evolutionarily closer to cynomolgus monkey and human P450 2J2 enzymes, than P450 2J forms in pigs, rabbits, rats or mice. 3. Marmoset P450 2J2 mRNA was abundantly expressed in the small intestine and liver, and to a lesser extent in the brain, lung and kidney. Immunoblot analysis also showed expression of marmoset P450 2J2 protein in the small intestine and liver. 4. Enzyme assays using marmoset P450 2J2 protein heterologously expressed in Escherichia coli indicated that marmoset P450 2J2 effectively catalyzed astemizole O-demethylation and terfenadine t-butyl hydroxylation, similar to human and cynomolgus monkey P450 2J2 enzymes. 5. These results suggest the functional characteristics of P450 2J2 enzymes are similar among marmosets, cynomolgus monkeys and humans.

    DOI: 10.3109/00498254.2016.1146366

    PubMed

  • Takahashi Tsuyoshi, Ohtsuka Tatsuyuki, Uno Yasuhiro, Utoh Masahiro, Yamazaki Hiroshi, Kume Toshiyuki .  Pre-incubation with cyclosporine A potentiates its inhibitory effects on pitavastatin uptake mediated by recombinantly expressed cynomolgus monkey hepatic organic anion transporting polypeptide. .  Biopharm Drug Dispos37 ( 8 ) 479 - 490   2016年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cyclosporine A, an inhibitor of hepatic organic anion transporting polypeptides (OATPs), reportedly increased plasma concentrations of probe substrates, although its maximum unbound blood concentrations were lower than the experimental half-maximal inhibitory (IC50 ) concentrations. Pre-incubation with cyclosporine A in vitro before simultaneous incubation with probes has been reported to potentiate its inhibitory effects on recombinant human OATP-mediated probe uptake. In the present study, the effects of cyclosporine A and rifampicin on recombinant cynomolgus monkey OATP-mediated pitavastatin uptake were investigated in pre- and simultaneous incubation systems. Pre-incubation with cyclosporine A, but not with rifampicin, decreased the apparent IC50 values on recombinant cynomolgus monkey OATP1B1- and OATP1B3-mediated pitavastatin uptake. Application of the co-incubated IC50 values toward R values (1 + [unbound inhibitor]inlet to the liver, theoretically maximum /inhibition constant) in static models, 1.1 in monkeys and 1.3 in humans, for recombinant cynomolgus monkey and human OATP1B1-mediated pitavastatin uptake might result in the poor prediction of drug interaction magnitudes. In contrast, the lowered IC50 values after pre-incubation with cyclosporine A provided better prediction with R values of 3.9 for monkeys and 2.7 for humans when the estimated maximum cyclosporine A concentrations at the inlet to the liver were used. These results suggest that the enhanced inhibitory potential of perpetrator medicines by pre- incubation on cynomolgus monkey OATP-mediated pitavastatin uptake in vitro could be of value for the precise estimation of drug interaction magnitudes in silico, in accordance with the findings from pre- administration of inhibitors on pitavastatin pharmacokinetics validated in monkeys. Copyright (c) 2016 John Wiley & Sons, Ltd.

    DOI: 10.1002/bdd.2039

    PubMed

  • Koyama Shuzo, Fukuda Koji, Watanabe Sho, Kohara Sakae, Tsuchiya Hideaki, Fukuzaki Koichiro, Nagano Masashi, Uno Yasuhiro, Hosoi Yoshihiko .  Development of a new device for artificial insemination in cynomolgus macaques. .  J Reprod Dev62 ( 5 ) 527 - 529   2016年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In cynomolgus macaques, an important animal species for biomedical research, efficient reproduction has been hampered partly due to the difficulties of artificial insemination (AI) using straw tubes developed for humans or farm animals, because cynomolgus macaques have a complex cervical canal structure. In this study, taking into consideration the unique structure of the macaque cervical canal, we developed a novel device for AI, comprised of a syringe and an outer cylinder. At 24 and 48 h after using this device to inject semen into one female, viable sperm were observed in the oviduct where the sperm meets the oocytes. We then attempted AI using this new device on 10 females that were at pre- ovulation, and pregnancy was successful in three animals (30% pregnancy rate). These results show that the newly developed device can be used for AI in cynomolgus macaques.

    DOI: 10.1262/jrd.2016-045

    PubMed

  • Igawa Yoshiyuki, Fujiwara Seiya, Ohura Kayoko, Hirokawa Takatsugu, Nishizawa You, Uehara Shotaro, Uno Yasuhiro, Imai Teruko .  Differences in Intestinal Hydrolytic Activities between Cynomolgus Monkeys and Humans: Evaluation of Substrate Specificities Using Recombinant Carboxylesterase 2 Isozymes. .  Mol Pharm13 ( 9 ) 3176 - 3186   2016年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys, used as an animal model to predict human pharmacokinetics, occasionally show different oral absorption patterns to humans due to differences in their intestinal metabolism. In this study, we investigated the differences between intestinal hydrolytic activities in cynomolgus monkeys and humans, in particular the catalyzing activities of their carboxylesterase 2 (CES2) isozymes. For this purpose we used both human and monkey microsomes and recombinant enzymes derived from a cell culture system. Monkey intestinal microsomes showed lower hydrolytic activity than human microsomes for several substrates. Interestingly, in contrast to human intestinal hydrolysis, which is not enantioselective, monkey intestine showed preferential R-form hydrolysis of propranolol derivatives. Recombinant CES2 isozymes from both species, mfCES2v3 from monkeys and human hCE2, showed similar metabolic properties to their intestinal microsomes when expressed in HEK293 cells. Recombinant hCE2 and mfCES2v3 showed similar Km values for both enantiomers of all propranolol derivatives tested. However, recombinant mfCES2v3 showed extreme R-enantioselective hydrolysis, and both hCE2 and mfCES2v3 showed lower activity for O-3-methyl-n-butyryl propranolol than for O-n-valeryl and O-2-methyl-n-butyryl propranolol. This lower hydrolytic activity was characterized by lower Vmax values. Docking simulations of the protein- ligand complex demonstrated that the enantioselectivity of mfCES2v3 for propranolol derivatives was possibly caused by the orientation of its active site being deformed by an amino acid change of Leu107 to Gln107 and the insertion of Met309, compared with hCE2. In addition, molecular dynamics simulation indicated the possibility that the interatomic distance between the catalytic triad and the substrate was elongated by a 3-positioned methyl in the propranolol derivatives. Overall, these findings will help us to understand the differences in intestinal hydrolytic activities between cynomolgus monkeys and humans.

    DOI: 10.1021/acs.molpharmaceut.6b00394

    PubMed

  • Iwasaki Kazuhide, Kitsugi Yusuke, Ikeda Kanami, Yoshikawa Takahiro, Hosaka Shinya, Uehara Shotaro, Uno Yasuhiro, Utoh Masahiro, Yamazaki Hiroshi .  In vivo individual variations in pharmacokinetics of efavirenz in cynomolgus monkeys genotyped for cytochrome P450 2C9. .  Biopharm Drug Dispos37 ( 6 ) 379 - 383   2016年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys are used frequently in preclinical studies for new drug development due to their evolutionary closeness to humans. An antiretroviral drug, efavirenz, is a typical probe substrate for human cytochrome P450 (P450) 2B6, but is mainly metabolized by cynomolgus monkey P450 2C9. In this study, plasma concentrations of efavirenz were assessed in six cynomolgus monkeys genotyped for P450 2C9 c.334 A > C (I112L) (three wild-type, one heterozygote and two homozygotes) by high performance liquid chromatography with tandem mass spectrometry. After intravenous administration at a dose of 1.0 mg/kg, biphasic plasma elimination curves of efavirenz were seen in these cynomolgus monkeys. The mean plasma concentration of the primary metabolite 8-hydroxyefavirenz (1 h after treatment, with hydrolysis by beta- glucuronidase) in the wild-type group was significantly higher (4.0-fold) than the combined heterozygous and homozygous group mean. The area under the plasma concentration-time curve value of efavirenz in the homozygous group after oral administration at a dose of 2.0 mg/kg was significantly higher (2.0-fold) than the combined wild-type and heterozygous group. These results collectively indicated that P450 2C9 c.334 A > C (I112L) variation was associated with efavirenz metabolic clearance in vivo. Cynomolgus P450 2C9 polymorphism might account for interindividual variations of efavirenz metabolism in cynomolgus monkeys. Copyright (c) 2016 John Wiley & Sons, Ltd.

    DOI: 10.1002/bdd.2021

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Kawano Mirai, Shimizu Makiko, Toda Akiko, Utoh Masahiro, Sasaki Erika, Yamazaki Hiroshi .  Individual Differences in Metabolic Clearance of S-Warfarin Efficiently Mediated by Polymorphic Marmoset Cytochrome P450 2C19 in Livers. .  Drug Metab Dispos44 ( 7 ) 911 - 915   2016年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Marmoset cytochrome P450 2C19, highly homologous to human P450 2C9 and 2C19, has been identified in common marmosets (Callithrix jacchus), a nonhuman primate species used in drug metabolism studies. Although genetic variants in human and macaque P450 2C genes account for the interindividual variability in drug metabolism, genetic variants have not been investigated in the marmoset P450 2C19 In this study, sequencing of P450 2C19 in 24 marmosets identified three variants p.[(Phe7Leu; Ser254Leu; Ile469Thr)], which showed substantially reduced metabolic capacity of S-warfarin compared with the wild-type group in vivo and in vitro. Although mean plasma concentrations of R-warfarin in marmosets determined after chiral separation were similar between the homozygous mutant and wild-type groups up to 24 hours after the intravenous and oral administrations of racemic warfarin, S-warfarin depletion from plasma was significantly faster in the three wild-type marmosets compared with the three homozygous mutant marmosets. These variants, cosegregating in the marmosets analyzed, influenced metabolic activities in 18 marmoset liver microsomes because the homozygotes and heterozygotes showed significantly reduced catalytic activities in liver microsomes toward S-warfarin 7-hydroxylation compared with the wild-type group. Kinetic analysis for S-warfarin 7-hydroxylation indicated that the recombinant P450 2C19 Ser254Leu variant would change the metabolic capacity. These results indicated that the interindividual variability of P450 2C-dependent drug metabolism such as S-warfarin clearance is at least partly accounted for by P450 2C19 variants in marmosets, suggesting that polymorphic P450 2C-dependent catalytic functions are relatively similar between marmosets and humans.

    DOI: 10.1124/dmd.116.070383

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Suzuki Takako, Utoh Masahiro, Sasaki Erika, Yamazaki Hiroshi .  Caffeine 7-N-demethylation and C-8-oxidation mediated by liver microsomal cytochrome P450 enzymes in common marmosets. .  Xenobiotica46 ( 7 ) 573 - 578   2016年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. 3-N-Demethylation of caffeine (1,3,7-trimethylxanthine) is mediated by human cytochrome P450 1A2, whereas 7-N-demethylation and C-8-hydroxylation are reportedly catalyzed by monkey P450 2C9 and rat P450 1A2, respectively. 2. Roles of marmoset P450 enzymes in caffeine oxidation were investigated using nine marmoset liver microsomes and 14 recombinantly expressed marmoset P450 enzymes. 3. Predominant caffeine 7-N-demethylation and C-8-hydroxylation activities in marmoset liver microsomes were moderately (r = 0.78, p < 0.05) and highly (r = 0.82, p < 0.01) correlated with midazolam 1'-hydroxylation activities, respectively, while the former was not strongly affected by ketoconazole or alpha-naphthoflavone. 4. Caffeine C-8-hydroxylation in liver microsomes was inhibited by ketoconazole and activated by alpha- naphthoflavone, suggesting main involvements of P450 3As. 5. Recombinant marmoset P450 3As had high Vmax/Km values for C-8-hydroxylation, comparable to Km values for marmoset liver microsomes. Marmoset P450 1As efficiently mediated caffeine 3-N-demethylation and C-8-hydroxylation with apparently lower Km values than those of liver microsomes. 6. These results collectively suggest highly active marmoset P450 3A enzymes toward caffeine 8-hydorxylaiton and involvement of multiple P450 isoforms including P450 1A in caffeine 7-N- and 3-N-demethylations in marmoset livers. Marmoset P450s have slightly different properties to human or monkey P450s regarding caffeine metabolic pathways.

    DOI: 10.3109/00498254.2015.1096980

    PubMed

  • Hosaka Shinya, Murayama Norie, Satsukawa Masahiro, Uehara Shotaro, Shimizu Makiko, Iwasaki Kazuhide, Iwano Shunsuke, Uno Yasuhiro, Yamazaki Hiroshi .  Identification of putative substrates for cynomolgus monkey cytochrome P450 2C8 by substrate depletion assays with 22 human P450 substrates and inhibitors. .  Biopharm Drug Dispos37 ( 5 ) 310 - 313   2016年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys are widely used in drug developmental stages as non- human primate models. Previous studies used 89 compounds to investigate species differences associated with cytochrome P450 (P450 or CYP) function that reported monkey specific CYP2C76 cleared 19 chemicals, and homologous CYP2C9 and CYP2C19 metabolized 17 and 30 human CYP2C9 and/or CYP2C19 substrates/inhibitors, respectively. In the present study, 22 compounds selected from viewpoints of global drug interaction guidances and guidelines were further evaluated to seek potential substrates for monkey CYP2C8, which is highly homologous to human CYP2C8 (92%). Amodiaquine, montelukast, quercetin and rosiglitazone, known as substrates or competitive inhibitors of human CYP2C8, were metabolically depleted by recombinant monkey CYP2C8 at relatively high rates. Taken together with our reported findings of the slow eliminations of amodiaquine and montelukast by monkey CYP2C9, CYP2C19 and CYP2C76, the present results suggest that these at least four chemicals may be good marker substrates for monkey CYP2C8. Copyright (c) 2016 John Wiley & Sons, Ltd.

    DOI: 10.1002/bdd.1998

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Yuki Yukako, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  A New Marmoset P450 4F12 Enzyme Expressed in Small Intestines and Livers Efficiently Metabolizes Antihistaminic Drug Ebastine. .  Drug Metab Dispos44 ( 6 ) 833 - 841   2016年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Common marmosets (Callithrix jacchus) are attracting attention as animal models in preclinical studies for drug development. However, cytochrome P450s (P450s), major drug-metabolizing enzymes, have not been fully identified and characterized in marmosets. In this study, based on the four novel P450 4F genes found on the marmoset genome, we successfully isolated P450 4F2, 4F3B, 4F11, and 4F12 cDNAs in marmoset livers. Deduced amino acid sequences of the four marmoset P450 4F forms exhibited high sequence identities (87%-93%) to the human and cynomolgus monkey P450 4F homologs. Marmoset P450 4F3B and 4F11 mRNAs were predominantly expressed in livers, whereas marmoset P450 4F2 and 4F12 mRNAs were highly expressed in small intestines and livers. Four marmoset P450 4F proteins heterologously expressed in Escherichia coli catalyzed the omega- hydroxylation of leukotriene B4 In addition, marmoset P450 4F12 effectively catalyzed the hydroxylation of antiallergy drug ebastine, a human P450 2J/4F probe substrate. Ebastine hydroxylation activities by small intestine and liver microsomes from marmosets and cynomolgus monkeys showed greatly higher values than those of humans. Ebastine hydroxylation activities by marmoset and cynomolgus monkey small intestine microsomes were inhibited (approximately 60%) by anti-P450 4F antibodies, unlike human small intestine microsomes, suggesting that contribution of P450 4F enzymes for ebastine hydroxylation in the small intestine might be different between marmosets/cynomolgus monkeys and humans. These results indicated that marmoset P450 4F2, 4F3B, 4F11, and 4F12 were expressed in livers and/or small intestines and were functional in the metabolism of endogenous and exogenous compounds, similar to those of cynomolgus monkeys and humans.

    DOI: 10.1124/dmd.116.070367

    PubMed

  • Sakai Chizuka, Iwano Shunsuke, Shimizu Makiko, Onodera Jun, Uchida Masashi, Sakurada Eri, Yamazaki Yuri, Asaoka Yoshiji, Imura Naoko, Uno Yasuhiro, Murayama Norie, Hayashi Ryoji, Yamazaki Hiroshi, Miyamoto Yohei .  Analysis of gene expression for microminipig liver transcriptomes using parallel long-read technology and short-read sequencing. .  Biopharm Drug Dispos37 ( 4 ) 220 - 232   2016年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The microminipig is one of the smallest minipigs that has emerged as a possible experimental animal model, because it shares many anatomical and/or physiological similarities with humans, including the coronary artery distribution in the heart, the digestive physiology, the kidney size and its structure, and so on. However, information on gene expression profiles, including those on drug-metabolizing phase I and II enzymes, in the microminipig is limited. Therefore, the aim of the present study was to identify transcripts in microminipig livers and to determine gene expression profiles. De novo assembly and expression analyses of microminipig transcripts were conducted with liver samples from three male and three female microminipigs using parallel long-read and short-read sequencing technologies. After unique sequences had been automatically aligned by assembling software, the mean contig length of 50843 transcripts was 707 bp. The expression profiles of cytochrome P450 (P450) 1A2, 2C, 2E1 and 3A genes in livers in microminipigs were similar to those in humans. Liver carboxylesterase (CES) precursor, liver CES- like, UDP-glucuronosyltransferase (UGT) 2C1-like, amine sulfotransferase (SULT)-like, N-acetyltransferases (NAT8) and glutathione S-transferase (GST) A2 genes, which are relatively unknown genes in pigs and/or humans, were expressed strongly. Furthermore, no significant gender differences were observed in the gene expression profiles of phase I enzymes, whereas UGT2B17, SULT1E1, SULT2A1, amine SULT-like, NAT8 and GSTT4 genes were different between males and females among phase II enzyme genes under the present sample conditions. These results provide a foundation for mechanistic studies and the use of microminipigs as model animals for drug development in the future. Copyright (c) 2016 John Wiley & Sons, Ltd.

    DOI: 10.1002/bdd.2007

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Molecular Cloning, Tissue Distribution, and Functional Characterization of Marmoset Cytochrome P450 1A1, 1A2, and 1B1. .  Drug Metab Dispos44 ( 1 ) 8 - 15   2016年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The common marmoset (Callithrix jacchus), a New World monkey, has potential to be an animal model for drug metabolism studies. In this study, we identified and characterized cytochrome P450 (P450) 1A1 and 1B1 in addition to the known P450 1A2 in marmosets. Marmoset P450 1A1 and 1B1 cDNA contained open reading frames encoding 512 and 543 amino acids, respectively, with high sequence identities (90%-93%) to other primate P450 1A1s and 1B1s. A phylogenetic tree based on amino acid sequences showed close evolutionary relationships among marmoset, macaque, and human P450 1A and 1B enzymes. By mRNA quantification and immunoblot analyses in five marmoset tissues, P450 1A1 was mainly expressed in lungs and small intestines, and P450 1A2 was expressed predominantly in livers. In contrast, P450 1B1 was expressed in all tissues tested. Marmoset P450 1A1, 1A2, and 1B1 heterologously expressed in Escherichia coli catalyzed 7-ethoxyresorufin O-deethylation, 7-ethoxycoumarin O-deethylation, and phenacetin O-deethylation, similar to those of humans and cynomolgus monkeys. Notably, marmoset P450 1A1 and 1A2 more efficiently catalyzed 7-ethoxyresorufin O-deethylation than those of the human homologs, but were comparable to those of the cynomolgus monkey homologs. Additionally, marmoset P450 1B1 preferentially catalyzed estradiol 4-hydroxylation; however, rat P450 1B1 more favorably catalyzed estradiol 2-hydroxylation, indicating that the estradiol hydroxylation specificity of marmoset P450 1B1 was similar to those of human and cynomolgus monkey P450 1B1. These results indicated that marmoset P450 1A and 1B enzymes had functional characteristics similar to those of humans and cynomolgus monkeys, suggesting that P450 1A and 1B-dependent metabolism was similar among marmosets, cynomolgus monkeys, and humans.

    DOI: 10.1124/dmd.115.067561

    PubMed

  • Uno Yasuhiro, Yamazaki Hiroshi .  Development of a genotyping tool for a functionally relevant CYP2C19 allele (Phe100Asn, Ala103Val and Ile112Leu) in cynomolgus macaques. .  J Vet Med Sci78 ( 1 ) 147 - 148   2016年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    .In cynomolgus macaques, which are widely used in drug metabolism studies, CYP2C19 (formerly known as CYP2C75) is abundantly expressed in liver, metabolizes human CYP2C substrates and is thus an important drug- metabolizing enzyme. One of the cynomolgus CYP2C19 alleles (p.Phe100Asn, p.Ala103Val and p.Ile112Leu) results in substantially reduced metabolic activity and thus is an important allele in drug metabolism studies. For this allele, a genotyping tool was developed using allele-specific TaqMan probe. Genotyping 40 Cambodian cynomolgus macaques using this tool found 1 homozygote, 17 heterozygotes and 22 wild type animals, and the result was confirmed by direct-sequencing. Interestingly, this allele frequency was similar to that of Chinese cynomolgus macaques. The genotyping tool established is useful for drug metabolism studies using cynomolgus macaques.

    DOI: 10.1292/jvms.15-0416

    PubMed

  • Uehara Shotaro, Inoue Takashi, Utoh Masahiro, Toda Akiko, Shimizu Makiko, Uno Yasuhiro, Sasaki Erika, Yamazaki Hiroshi .  Simultaneous pharmacokinetics evaluation of human cytochrome P450 probes, caffeine, warfarin, omeprazole, metoprolol and midazolam, in common marmosets (Callithrix jacchus). .  Xenobiotica46 ( 2 ) 163 - 168   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Pharmacokinetics of human cytochrome P450 probes (caffeine, racemic warfarin, omeprazole, metoprolol and midazolam) composite, after single intravenous and oral administrations at doses of 0.20 and 1.0 mg kg(-1), respectively, to four male common marmosets were investigated. 2. The plasma concentrations of caffeine and warfarin decreased slowly in a monophasic manner but those of omeprazole, metoprolol and midazolam decreased extensively after intravenous and oral administrations, in a manner that approximated those as reported for pharmacokinetics in humans. 3. Bioavailabilities were approximately 100% for caffeine and warfarin, but <25% for omeprazole and metoprolol. Bioavailability of midazolam was 4% in marmosets, presumably because of contribution of marmoset P450 3A4 expressed in small intestine and liver, with a high catalytic efficiency for midazolam 1'-hydroxylation as evident in the recombinant system. 4. These results suggest that common marmosets, despite their rapid clearance of some human P450 probe substrates, could be an experimental model for humans and that marmoset P450s have functional characteristics that differ from those of human and/or cynomolgus monkey P450s in some aspects, indicating their importance in modeling in P450-dependent drug metabolism studies in marmosets and of further studies.

    DOI: 10.3109/00498254.2015.1057270

    PubMed

  • Hosaka Shinya, Murayama Norie, Satsukawa Masahiro, Uehara Shotaro, Shimizu Makiko, Iwasaki Kazuhide, Iwano Shunsuke, Uno Yasuhiro, Yamazaki Hiroshi .  Similar substrate specificity of cynomolgus monkey cytochrome P450 2C19 to reported human P450 2C counterpart enzymes by evaluation of 89 drug clearances. .  Biopharm Drug Dispos36 ( 9 ) 636 - 643   2015年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys are used widely in preclinical studies as non-human primate species. The amino acid sequence of cynomolgus monkey cytochrome P450 (P450 or CYP) 2C19 is reportedly highly correlated to that of human CYP2C19 (92%) and CYP2C9 (93%). In the present study, 89 commercially available compounds were screened to find potential substrates for cynomolgus monkey CYP2C19. Of 89 drugs, 34 were metabolically depleted by cynomolgus monkey CYP2C19 with relatively high rates. Among them, 30 compounds have been reported as substrates or inhibitors of, either or both, human CYP2C19 and CYP2C9. Several compounds, including loratadine, showed high selectivity to cynomolgus monkey CYP2C19, and all of these have been reported as human CYP2C19 and/or CYP2C9 substrates. In addition, cynomolgus monkey CYP2C19 formed the same loratadine metabolite as human CYP2C19, descarboethoxyloratadine. These results suggest that cynomolgus monkey CYP2C19 is generally similar to human CYP2C19 and CYP2C9 in its substrate recognition functionality.

    DOI: 10.1002/bdd.1991

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Kawano Mirai, Shimizu Makiko, Toda Akiko, Utoh Masahiro, Sasaki Erika, Yamazaki Hiroshi .  Novel Marmoset Cytochrome P450 2C19 in Livers Efficiently Metabolizes Human P450 2C9 and 2C19 Substrates, S-Warfarin, Tolbutamide, Flurbiprofen, and Omeprazole. .  Drug Metab Dispos43 ( 10 ) 1408 - 1416   2015年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The common marmoset (Callithrix jacchus), a small New World monkey, has the potential for use in human drug development due to its evolutionary closeness to humans. Four novel cDNAs, encoding cytochrome P450 (P450) 2C18, 2C19, 2C58, and 2C76, were cloned from marmoset livers to characterize P450 2C molecular properties, including previously reported P450 2C8. The deduced amino acid sequence showed high sequence identities (>86%) with those of human P450 2Cs, except for marmoset P450 2C76, which has a low sequence identity ( approximately 70%) with any human P450 2Cs. Phylogenetic analysis showed that marmoset P450 2Cs were more closely clustered with those of humans and macaques than other species investigated. Quantitative polymerase chain reaction analysis showed that all of the marmoset P450 2C mRNAs were predominantly expressed in liver as opposed to the other tissues tested. Marmoset P450 2C proteins were detected in liver by immunoblotting using antibodies against human P450 2Cs. Among marmoset P450 2Cs heterologously expressed in Escherichia coli, marmoset P450 2C19 efficiently catalyzed human P450 2C substrates, S-warfarin, diclofenac, tolbutamide, flurbiprofen, and omeprazole. Marmoset P450 2C19 had high Vmax and low Km values for S-warfarin 7-hydroxylation that were comparable to those in human liver microsomes, indicating warfarin stereoselectivity similar to findings in humans. Faster in vivo S-warfarin clearance than R-warfarin after intravenous administration of racemic warfarin (0.2 mg/kg) to marmosets was consistent with the in vitro kinetic parameters. These results indicated that marmoset P450 2C enzymes had functional characteristics similar to those of humans, and that P450 2C-dependent metabolic properties are likewise similar between marmosets and humans.

    DOI: 10.1124/dmd.115.066100

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Substrate Selectivities and Catalytic Activities of Marmoset Liver Cytochrome P450 2A6 Differed from Those of Human P450 2A6. .  Drug Metab Dispos43 ( 7 ) 969 - 976   2015年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The common marmoset (Callithrix jacchus), a New World primate species, is potentially a useful animal model for preclinical studies in drug development. However, cytochrome P450 (P450) enzymes have not been fully identified and characterized in marmosets. In this study, we identified P450 2A6 cDNA with the sequence highly identical (91-94%) to human P450 2A6, 2A7, and 2A13 cDNA and cynomolgus monkey P450 2A23, 2A24, and 2A26 cDNA. Among the tissue types examined, marmoset P450 2A6 mRNA was most abundantly expressed in livers where P450 2A6 protein was also detected by immunoblotting. Phylogenetic analysis showed that marmoset P450 2A6 was more closely clustered with human and cynomolgus monkey P450 2As than P450 2As of dog, rat, and mouse (the species also used in drug metabolism). Marmoset P450 2A6 heterologously expressed in Escherichia coli membranes efficiently catalyzed 7-ethoxycoumarin O-deethylation, similar to human P450 2A6 and 2A13 and cynomolgus monkey P450 2A23, 2A24, and 2A26, but much less effectively coumarin 7-hydroxylation, showing some difference as well. Interestingly, marmoset P450 2A6 and cynomolgus monkey P450 2A23 catalyzed phenacetin O-deethylation, which is catalyzed by human P450 1A2 and 2A13, but not by P450 2A6. Marmoset P450 2A6 also exhibited catalytic activity toward testosterone by the multiple sites, but not rat P450 2A-specific testosterone 7alpha-hydroxylation activity. These results indicated that marmoset P450 2A6 had functional characteristics different from those of human and cynomolgus monkey P450 2As in terms of partially different substrate specificities and catalytic activities, indicating its importance of further studies for P450 2A-dependent drug metabolism in marmosets.

    DOI: 10.1124/dmd.115.063909

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Kohara Sakae, Osada Naoki, Murayama Norie, Yamazaki Hiroshi .  CYP2D44 polymorphisms in cynomolgus and rhesus macaques. .  Mol Biol Rep42 ( 7 ) 1149 - 1155   2015年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Macaques, including cynomolgus and rhesus macaques, are important animal species used in drug metabolism studies. CYP2D44 is expressed in cynomolgus macaque liver and encodes a functional drug metabolizing enzyme, metabolizing typical human CYP2D substrates such as bufuralol and dextromethorphan. CYP2D44 is highly homologous to human CYP2D6 that is known to be polymorphic with a large inter-individual variation in metabolic activities, however, genetic polymorphisms have not been investigated in macaque CYP2D44. In the present study, screening of 78 cynomolgus and 40 rhesus macaques found a total of 67 variants, including 64 non-synonymous variants, 1 nonsense mutation, and 2 frameshift mutations, and 1 gene conversion, of which 14, 19, and 15 variants were unique to Indochinese cynomolgus macaques, Indonesian cynomolgus macaques, and Chinese rhesus macaques, respectively. Eleven of the 64 non-synonymous variants were located in substrate recognition sites, the regions important for protein function. By site-directed mutagenesis and metabolic assays, S175N, V185L, A235G, R242G, R245K, and N337D showed substantially decreased activity in bufuralol 1'-hydroxylation as compared with wild-type proteins. Moreover, two null alleles (c.128T>del and c.664G>T) were found in Indonesian cynomolgus macaques, but not in Indochinese cynomolgus macaques or Chinese rhesus macaques. These results suggest that genetic polymorphisms might account for the variability of CYP2D44-dependent metabolism in macaques.

    DOI: 10.1007/s11033-015-3863-0

    PubMed

  • Hosaka Shinya, Murayama Norie, Satsukawa Masahiro, Uehara Shotaro, Shimizu Makiko, Iwasaki Kazuhide, Iwano Shunsuke, Uno Yasuhiro, Yamazaki Hiroshi .  Comprehensive Evaluation for Substrate Selectivity of Cynomolgus Monkey Cytochrome P450 2C9, a New Efavirenz Oxidase. .  Drug Metab Dispos43 ( 7 ) 1119 - 1122   2015年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys are widely used as primate models in preclinical studies, because of their evolutionary closeness to humans. In humans, the cytochrome P450 (P450) 2C enzymes are important drug-metabolizing enzymes and highly expressed in livers. The CYP2C enzymes, including CYP2C9, are also expressed abundantly in cynomolgus monkey liver and metabolize some endogenous and exogenous substances like testosterone, S-mephenytoin, and diclofenac. However, comprehensive evaluation regarding substrate specificity of monkey CYP2C9 has not been conducted. In the present study, 89 commercially available drugs were examined to find potential monkey CYP2C9 substrates. Among the compounds screened, 20 drugs were metabolized by monkey CYP2C9 at a relatively high rates. Seventeen of these compounds were substrates or inhibitors of human CYP2C9 or CYP2C19, whereas three drugs were not, indicating that substrate specificity of monkey CYP2C9 resembled those of human CYP2C9 or CYP2C19, with some differences in substrate specificities. Although efavirenz is known as a marker substrate for human CYP2B6, efavirenz was not oxidized by CYP2B6 but by CYP2C9 in monkeys. Liquid chromatography- mass spectrometry analysis revealed that monkey CYP2C9 and human CYP2B6 formed the same mono- and di-oxidized metabolites of efavirenz at 8 and 14 positions. These results suggest that the efavirenz 8-oxidation could be one of the selective markers for cynomolgus monkey CYP2C9 among the major three CYP2C enzymes tested. Therefore, monkey CYP2C9 has the possibility of contributing to limited specific differences in drug oxidative metabolism between cynomolgus monkeys and humans.

    DOI: 10.1124/dmd.115.063925

    PubMed

  • Uno Yasuhiro, Hosokawa Masakiyo, Imai Teruko .  Isolation and characterization of arylacetamide deacetylase in cynomolgus macaques. .  J Vet Med Sci77 ( 6 ) 721 - 724   2015年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Arylacetamide deacetylase (AADAC), a microsomal serine esterase, hydrolyzes drugs, such as flutamide, phenacetin and rifampicin. Because AADAC has not been fully investigated at molecular levels in cynomolgus macaques, the non-human primate species widely used in drug metabolism studies, cynomolgus AADAC cDNA was isolated and characterized. The deduced amino acid sequence, highly homologous (92%) to human AADAC, was more closely clustered with human AADAC than the dog, rat or mouse ortholog in a phylogenetic tree. AADAC was flanked by AADACL2 and SUCNR1 in the cynomolgus and human genomes. Moreover, relatively abundant expression of AADAC mRNA was found in liver and jejunum, the drug- metabolizing organs, in cynomolgus macaques, similar to humans. The results suggest molecular similarities of AADAC between cynomolgus macaques and humans.

    DOI: 10.1292/jvms.14-0496

    PubMed

  • Utoh Masahiro, Yoshikawa Takahiro, Hayashi Yoshiharu, Shimizu Makiko, Iwasaki Kazuhide, Uno Yasuhiro, Yamazaki Hiroshi .  Slow R-warfarin 7-hydroxylation mediated by P450 2C19 genetic variants in cynomolgus monkeys in vivo. .  Biochem Pharmacol95 ( 2 ) 110 - 114   2015年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys are widely used as non-human primate species in preclinical studies, due to their close evolutionary relationship to humans. Monkey cytochrome P450 2C19 (formerly known as P450 2C75), highly homologous to human P450 2C19, has been identified to be R-warfarin 7-hydroxylase in cynomolgus monkeys. In the present study, the in vivo pharmacokinetics of stereoselective warfarin and metabolites at a dose of 1.0mg/kg were investigated after oral and intravenous administration of racemic warfarin to fasted male cynomolgus monkeys (n=11, from Indochina, 4-8 years of age, 3.5-7.4kg of body weight), which had been genotyped for P450 2C19 [c.298TT>AA; c.308C>T; and c.334ATC>CTT]. Kinetic parameters for S-warfarin were not different among the homozygous mutant, heterozygous mutant, and wild type groups; however, values of elimination half-lives, area under the curves, and total body clearance of R-warfarin in the homozygous mutant group showed one-order differences from those values in the wild type group after oral or intravenous administration. R-Warfarin 7-hydroxylations in vivo in homozygous mutant groups were slow compared to wild type or heterozygous mutant groups. These results demonstrate that inter-animal variations of R-warfarin clearance in cynomolgus monkeys are associated with P450 2C19 genetic variants [p.Phe100Asn, p.Ala103Val, and p.Ile112Leu]. Because some interindividual variability of P450 2C-dependent drug metabolism in cynomolgus monkeys, similarly in humans, is accounted for by polymorphic P450 2C19 variants, genotyping of drug metabolism enzymes should be considered before and after P450-dependent drug metabolism testing and evaluations in cynomolgus monkeys.

    DOI: 10.1016/j.bcp.2015.03.008

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Inoue Takashi, Murayama Norie, Shimizu Makiko, Sasaki Erika, Yamazaki Hiroshi .  Activation and deactivation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine by cytochrome P450 enzymes and flavin-containing monooxygenases in common marmosets (Callithrix jacchus). .  Drug Metab Dispos43 ( 5 ) 735 - 742   2015年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The potential proneurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induces Parkinson-like syndromes in common marmosets, other primates, and humans. MPTP is metabolically activated to 1-methyl-4-phenyl-2,3-dihydropyridinium and 1-methyl-4-phenylpyridinium ions (MPDP(+) and MPP(+), respectively) by desaturation reactions. MPTP is deactivated to 4-phenyl-1,2,3,6-tetrahydropyridine (PTP) by N-demethylation and is also deactivated to MPTP N-oxide. The roles of cytochrome P450 (P450) enzymes and flavin-containing monooxygenases (FMOs) in the oxidative metabolism of MPTP-treated marmosets are not yet fully clarified. This study aimed to elucidate P450- and FMO-dependent MPTP metabolism in marmoset liver and brain. Rates of MPTP N-oxygenation in liver microsomes were similar to those in brain microsomes from 11 individual marmosets (substrate concentration, 50 muM) and were correlated with rates of benzydamine N-oxygenation (r = 0.75, P < 0.05); the reactions were inhibited by methimazole (10 muM). MPTP N-oxygenation was efficiently mediated by recombinantly expressed marmoset FMO3. Rates of PTP formation by MPTP N-demethylation in marmoset liver microsomes were correlated with bufuralol 1'-hydroxylation rates (r = 0.77, P < 0.01) and were suppressed by quinidine (1 muM), thereby indicating the importance of marmoset CYP2D6 in PTP formation. MPTP transformations to MPDP(+) and MPP(+) were efficiently catalyzed by recombinant marmoset CYP2D6 and human CYP1A2. These results indicated the contributions of multiple drug-metabolizing enzymes to MPTP oxidation, especially marmoset FMO3 in deactivation (N-oxygenation) and marmoset CYP2D6 for both MPTP deactivation and MPTP activation to MPDP(+) and MPP(+). These findings provide a foundation for understanding MPTP metabolism and for the successful production of preclinical marmoset models.

    DOI: 10.1124/dmd.115.063594

    PubMed

  • Koyanagi Takashi, Nakanishi Yasuharu, Murayama Norie, Yamaura Yoshiyuki, Ikeda Kanami, Yano Koji, Uehara Shotaro, Utoh Masahiro, Kim Soonih, Uno Yasuhiro, Yamazaki Hiroshi .  Age-related changes of hepatic clearances of cytochrome P450 probes, midazolam and R-/S-warfarin in combination with caffeine, omeprazole and metoprolol in cynomolgus monkeys using in vitro-in vivo correlation. .  Xenobiotica45 ( 4 ) 312 - 321   2015年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Pharmacokinetics of human cytochrome P450 probes (caffeine, racemic warfarin, omeprazole, metoprolol and midazolam) were investigated after single intravenous and oral administrations at doses of 0.20 and 1.0 mg kg(-1), respectively, in combination to three young (3-year-old) and three aged (16-year-old) cynomolgus monkeys. 2. The plasma concentrations of caffeine and R-/S-warfarin decreased slowly in a monophasic manner, but those of omeprazole, metoprolol and midazolam decreased rapidly, in a similar manner to those as reported for pharmacokinetics in humans. 3. The mean maximum concentrations of R- and S-warfarin (4.6 and 3.7 microg/mL, respectively) in aged monkeys after oral administration were significantly higher than those in young monkeys (3.3 and 2.7 microg/mL). The mean clearance (CL) values of midazolam in aged monkeys (9.5 mL/min/kg) were significantly lower than those in young monkeys (13 mL/min/kg). 4. Individual intrinsic CL values for omeprazole (r = 0.29) and metoprolol (r = 0.30) of individual monkey livers were inversely correlated with their ages significantly (p < 0.05) in liver microsomes prepared from 55 cynomolgus monkeys. 5. These results suggest that cynomolgus monkeys could be a good model for humans, especially with particular characteristics in reduced CLs of some human P450 substrates by aging.

    DOI: 10.3109/00498254.2014.979271

    PubMed

  • Uehara Shotaro, Murayama Norie, Nakanishi Yasuharu, Nakamura Chika, Hashizume Takanori, Zeldin Darryl C, Yamazaki Hiroshi, Uno Yasuhiro .  Immunochemical quantification of cynomolgus CYP2J2, CYP4A and CYP4F enzymes in liver and small intestine. .  Xenobiotica45 ( 2 ) 124 - 130   2015年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. An increasing number of studies have indicated the roles of CYP4 proteins in drug metabolism; however, CYP4 expression has not been measured in cynomolgus monkeys, an important animal species for drug metabolism studies. 2. In this study, cynomolgus CYP4A11, CYP4F2/3, CYP4F11 and CYP4F12, along with CYP2J2, were immunoquantified using selective antibodies in 28 livers and 35 small intestines, and their content was compared with CYP1A, CYP2A, CYP2B6, CYP2C9/19, CYP2D, CYP2E1, CYP3A4 and CYP3A5, previously quantified. 3. In livers, CYP2J2, CYP4A11, CYP4F2/3, CYP4F11 and CYP4F12, varied 1.3- to 4.3-fold, represented 11.2, 14.4, 8.0, 2.7 and 0.3% of total immunoquantified CYP1-4 proteins, respectively. 4. In small intestines, CYP2J2, CYP4F2/3, CYP4F11 and CYP4F12, varied 2.4- to 9.7-fold, represented 6.9, 36.4, 2.4 and 9.3% of total immunoquantified CYP1-4 proteins, respectively, making CYP4F the most abundant P450 subfamily in small intestines. CYP4A11 was under the detection limit in all of the samples analyzed. 5. Significant correlations were found in liver for CYP4A11 with lauric acid 11-/12-hydroxylation and for CYP4F2/3 and CYP4F11 with astemizole hydroxylation. 6. This study revealed the relatively abundant contents of cynomolgus CYP2J2, CYP4A11 and CYP4Fs in liver and/or small intestine, suggesting their potential roles for the metabolism of xenobitotics and endogenous substrates.

    DOI: 10.3109/00498254.2014.952800

    PubMed

  • Uno Yasuhiro, Matsushita Akinori, Murayama Norie, Yamazaki Hiroshi .  Genetic polymorphism of cynomolgus and rhesus macaque CYP2C9. .  Drug Metab Pharmacokinet30 ( 1 ) 130 - 132   2015年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus and rhesus macaques are non-human primate species widely used in drug metabolism studies. Cynomolgus CYP2C9 (formerly known as CYP2C43) is predominantly expressed in liver and encodes a drug-metabolizing enzyme that metabolizes human CYP2C substrates such as S-mephenytoin and progesterone. In addition, cynomolgus CYP2C9 also metabolizes caffeine, resulting in the formation of the metabolite that is not generated efficiently in humans. Genetic variants of human CYP2C genes account for the inter-individual variability in drug metabolism: however, CYP2C9 variants have not been found in macaques. To see if CYP2C9 is polymorphic in macaques, in this study, CYP2C9 was re-sequenced in 78 cynomolgus and 36 rhesus macaques. A total of 27 non-synonymous variants were found, among which 4 were located in substrate recognition sites, the domain important for protein function. Thirteen and seven variants were unique to cynomolgus and rhesus macaques, respectively. This study revealed the polymorphic nature of cynomolgus and rhesus CYP2C9, similar to human CYP2C genes, by identification of numerous genetic variants including non-synonymous variants.

    DOI: 10.1016/j.dmpk.2014.10.002

    PubMed

  • Hosaka Shinya, Murayama Norie, Satsukawa Masahiro, Shimizu Makiko, Uehara Shotaro, Fujino Hideki, Iwasaki Kazuhide, Iwano Shunsuke, Uno Yasuhiro, Yamazaki Hiroshi .  Evaluation of 89 compounds for identification of substrates for cynomolgus monkey CYP2C76, a new bupropion/nifedipine oxidase. .  Drug Metab Dispos43 ( 1 ) 27 - 33   2015年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys are widely used in preclinical studies during drug development because of their evolutionary closeness to humans, including their cytochrome P450s (P450s). Most cynomolgus monkey P450s are almost identical (>/=90%) to human P450s; however, CYP2C76 has low sequence identity (approximately 80%) to any human CYP2Cs. Although CYP2C76 has no ortholog in humans and is partly responsible for species differences in drug metabolism between cynomolgus monkeys and humans, a broad evaluation of potential substrates for CYP2C76 has not yet been conducted. In this study, a screening of 89 marketed compounds, including human CYP2C and non-CYP2C substrates or inhibitors, was conducted to find potential CYP2C76 substrates. Among the compounds screened, 19 chemicals were identified as substrates for CYP2C76, including substrates for human CYP1A2 (7-ethoxyresorufin), CYP2B6 (bupropion), CYP2D6 (dextromethorphan), and CYP3A4/5 (dextromethorphan and nifedipine), and inhibitors for CYP2B6 (sertraline, clopidogrel, and ticlopidine), CYP2C8 (quercetin), CYP2C19 (ticlopidine and nootkatone), and CYP3A4/5 (troleandomycin). CYP2C76 metabolized a wide variety of the compounds with diverse structures. Among them, bupropion and nifedipine showed high selectivity to CYP2C76. As for nifedipine, CYP2C76 formed methylhydroxylated nifedipine, which was not produced by monkey CYP2C9, CYP2C19, or CYP3A4, as identified by mass spectrometry and estimated by a molecular docking simulation. This unique oxidative metabolite formation of nifedipine could be one of the selective marker reactions of CYP2C76 among the major CYP2Cs and CYP3As tested. These results suggest that monkey CYP2C76 contributes to bupropion hydroxylation and formation of different nifedipine oxidative metabolites as a result of its relatively large substrate cavity.

    DOI: 10.1124/dmd.114.061275

    PubMed

  • Okamatsu Gaku, Komatsu Tetsuya, Kubota Akira, Onaga Takenori, Uchide Tsuyoshi, Endo Daiji, Kirisawa Rikio, Yin Guojun, Inoue Hiroki, Kitazawa Takio, Uno Yasuhiro, Teraoka Hiroki .  Identification and functional characterization of novel feline cytochrome P450 2A. .  Xenobiotica45 ( 6 ) 503 - 510   2015年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Cytochrome P450s are the major metabolizing enzymes for xenobiotics in humans and other mammals. Although the domestic cat Felis catus, an obligate carnivore, is the most common companion animal, the properties of cytochrome P450 subfamilies are largely unknown. 2. We newly identified the feline CYP2A13, which consists of 494 deduced amino acids, showing the highest identity to CYP2As of dogs, followed by those of pigs, cattle and humans. 3. The feline CYP2A13 transcript and protein were expressed almost exclusively in the liver without particular sex- dependent differences. 4. The feline CYP2A13 protein heterogeneously expressed in Escherichia coli showed metabolic activity similar to those of human and canine CYP2As for coumarin, 7-ethoxycoumarin and nicotine. 5. The results indicate the importance of CYP2A13 in systemic metabolism of xenobiotics in cats.

    DOI: 10.3109/00498254.2014.998322

    PubMed

  • Uehara Shotaro, Uno Yasuhiro, Hagihira Yuya, Murayama Norie, Shimizu Makiko, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi .  Marmoset cytochrome P450 2D8 in livers and small intestines metabolizes typical human P450 2D6 substrates, metoprolol, bufuralol and dextromethorphan. .  Xenobiotica45 ( 9 ) 766 - 772   2015年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Although the New World non-human primate, the common marmoset (Callithrix jacchus), is a potentially useful animal model, comprehensive understanding of drug metabolizing enzymes is insufficient. 2. A cDNA encoding a novel cytochrome P450 (P450) 2D8 was identified in marmosets. The amino acid sequence deduced from P450 2D8 cDNA showed a high sequence identity (83-86%) with other primate P450 2Ds. Phylogenetic analysis showed that marmoset P450 2D8 was closely clustered with human P450 2D6, unlike P450 2Ds of miniature pig, dog, rabbit, guinea pig, mouse or rat. 3. Marmoset P450 2D8 mRNA was predominantly expressed in the liver and small intestine among the tissues types analyzed, whereas marmoset P450 2D6 mRNA was expressed predominantly in the liver where P450 2D protein was detected by immunoblotting. 4. By metabolic assays using marmoset P450 2D8 protein heterologously expressed in Escherichia coli, although P450 2D8 exhibits lower catalytic efficiency compared to marmoset and human P450 2D6 enzymes, P450 2D8 mediated O-demethylations of metoprolol and dextromethorphan and bufuralol 1'-hydroxylation. 5. These results suggest that marmoset P450 2D8 (also expressed in the extrahepatic tissues) has potential roles in drug metabolism in a similar manner to those of human and marmoset P450 2D6.

    DOI: 10.3109/00498254.2015.1019595

    PubMed

  • Shida Satomi, Utoh Masahiro, Murayama Norie, Shimizu Makiko, Uno Yasuhiro, Yamazaki Hiroshi .  Human plasma concentrations of cytochrome P450 probes extrapolated from pharmacokinetics in cynomolgus monkeys using physiologically based pharmacokinetic modeling. .  Xenobiotica45 ( 10 ) 881 - 886   2015年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    1. Cynomolgus monkeys are widely used in preclinical studies as non-human primate species. Pharmacokinetics of human cytochrome P450 probes determined in cynomolgus monkeys after single oral or intravenous administrations were extrapolated to give human plasma concentrations. 2. Plasma concentrations of slowly eliminated caffeine and R-/S-warfarin and rapidly eliminated omeprazole and midazolam previously observed in cynomolgus monkeys were scaled to human oral biomonitoring equivalents using known species allometric scaling factors and in vitro metabolic clearance data with a simple physiologically based pharmacokinetic (PBPK) model. Results of the simplified human PBPK models were consistent with reported experimental PK data in humans or with values simulated by a fully constructed population-based simulator (Simcyp). 3. Oral administrations of metoprolol and dextromethorphan (human P450 2D probes) in monkeys reportedly yielded plasma concentrations similar to their quantitative detection limits. Consequently, ratios of in vitro hepatic intrinsic clearances of metoprolol and dextromethorphan determined in monkeys and humans were used with simplified PBPK models to extrapolate intravenous PK in monkeys to oral PK in humans. 4. These results suggest that cynomolgus monkeys, despite their rapid clearance of some human P450 substrates, could be a suitable model for humans, especially when used in conjunction with simple PBPK models.

    DOI: 10.3109/00498254.2015.1028511

    PubMed

  • Uno Yasuhiro, Hosaka Shinya, Yamazaki Hiroshi .  Identification and analysis of CYP7A1, CYP17A1, CYP20A1, CYP27A1 and CYP51A1 in cynomolgus macaques. .  J Vet Med Sci76 ( 12 ) 1647 - 1650   2014年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cytochromes P450 (P450) are important for not only drug metabolism and toxicity, but also biosynthesis and metabolism of cholesterol and bile acids, and steroid synthesis. In cynomolgus macaques, widely used in biomedical research, we have characterized P450 cDNAs, which were isolated as expressed sequence tags of cynomolgus macaque liver. In this study, cynomolgus CYP7A1, CYP17A1, CYP20A1, CYP27A1 and CYP51A1 cDNAs were characterized by sequence analysis, phylogenetic analysis and tissue expression pattern. By sequence analysis, these five cynomolgus P450s had high sequence identities (94-99%) to the human orthologs in amino acids. By phylogenetic analysis, each cynomolgus P450 was more closely related to the human ortholog as compared with the dog or rat ortholog. By quantitative polymerase chain reaction, among the 10 tissue types, CYP7A1 and CYP17A1 mRNAs were preferentially expressed in liver and adrenal gland, respectively. Cynomolgus CYP27A1 and CYP51A1 mRNAs were most abundantly expressed in liver and testis, respectively. Cynomolgus CYP20A1 mRNA was expressed in all the tissues, including brain and liver. Tissue expression patterns of each cynomolgus P450 were generally similar to that of the human ortholog. These results suggest the molecular similarities of CYP7A1, CYP17A1, CYP20A1, CYP27A1 and CYP51A1 between cynomolgus macaques and humans.

    DOI: 10.1292/jvms.14-0313

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Hosokawa Masakiyo, Imai Teruko .  Systematic identification and characterization of carboxylesterases in cynomolgus macaques. .  Drug Metab Dispos42 ( 12 ) 2002 - 2006   2014年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Carboxylesterase (CES) is important for detoxification of a wide range of drugs and xenobiotics and catalyzes cholesterol and fatty acid metabolism. Cynomolgus macaques are widely used in drug metabolism studies; however, cynomolgus CES has not been fully investigated at molecular levels, partly due to the lack of gene information. In this study, we isolated and characterized cDNAs for CES homologous to human CES1, CES2, and CES5A in cynomolgus macaques. By genome analysis, in the cynomolgus macaque genome, three gene sequences were found for CES1(v1-3) and CES2(v1-3), whereas one gene sequence was found for CES5A. Cynomolgus CES1, CES2, and CES5A genes were located in the genomic regions corresponding to the human genes. We successfully identified CES1v1, CES1v2, CES2v1, CES2v3, and CES5A cDNAs from cynomolgus liver. Sequence analysis showed that amino acid sequences of each CES were highly homologous to that of the human homolog. All five CESs had sequences characteristic for CES enzymes, including the catalytic triad and oxyanion hole loop. By quantitative polymerase chain reaction, the most abundant expression of CES mRNAs among the 10 tissue types analyzed was observed in liver (CES1v1 and CES2v3 mRNAs), jejunum (CES2v1 mRNAs), and kidney (CES1v2 and CES5A mRNA), the organs important for drug metabolism and excretion. The results indicated that cynomolgus macaques express at least five CES genes, which potentially encode intact CES proteins.

    DOI: 10.1124/dmd.114.059972

    PubMed

  • Uno Yasuhiro, Matsushita Akinori, Shukuya Mitsunori, Matsumoto Yasuka, Murayama Norie, Yamazaki Hiroshi .  CYP2C19 polymorphisms account for inter-individual variability of drug metabolism in cynomolgus macaques. .  Biochem Pharmacol91 ( 2 ) 242 - 248   2014年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    CYP2C19 (formerly known as CYP2C75), highly homologous to human CYP2C19, has been identified in cynomolgus and rhesus macaques, non-human primate species widely used in drug metabolism studies. CYP2C19 is predominantly expressed in liver and encodes a functional drug-metabolizing enzyme. Genetic variants in human CYP2C genes account for the inter-individual variability in drug metabolism; however, genetic variants have not been investigated in macaque CYP2C19. In the present study, re-sequencing of CYP2C19 in 78 cynomolgus and 36 rhesus macaques identified 34 non- synonymous variants. Among these, 6 were located in substrate recognition sites, the domains important for protein function. Eighteen and 6 variants were unique to cynomolgus and rhesus macaques, respectively. Four variants were characterized by site-directed mutagenesis and metabolic assays, and 3 variants (p.Phe100Asn, p.Ala103Val, and p.Ile112Leu) showed substantially reduced activity as compared with wild type in flurbiprofen 4'-hydroxylation, omeprazole 5-hydroxylation, and R-/S-warfarin 7-hydroxylation. These variants, co-segregating in the animals analyzed, influenced metabolic activities because the homozygotes and/or heterozygotes showed significantly reduced catalytic activities in liver toward flurbiprofen 4'-hydroxylation and omeprazole 5-hydroxylation as compared with wild type. Kinetic analysis for R-warfarin 7-hydroxylation and docking simulation indicated that CYP2C19 Ala103Val would change the function and conformation of this enzyme. Ala103Val variation diminished homotropic cooperativity of CYP2C19 with R-warfarin yielding low metabolic capacity. These results indicated that the interindividual variability of CYP2C-dependent drug metabolism is at least partly accounted for by CYP2C19 variants in cynomolgus macaques.

    DOI: 10.1016/j.bcp.2014.07.004

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Kohara Sakae, Murayama Norie, Yamazaki Hiroshi .  Polymorphisms of CYP2D17 in cynomolgus and rhesus macaques: an evidence of the genetic basis for the variability of CYP2D-dependent drug metabolism. .  Drug Metab Dispos42 ( 9 ) 1407 - 1410   2014年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus macaques and rhesus macaques are nonhuman primate species widely used in drug metabolism studies. Cynomolgus CYP2D17, highly homologous to human CYP2D6, metabolizes human CYP2D6 substrates such as bufuralol and dextromethorphan, and the gene is expressed predominantly in liver. Although human CYP2D6 variants account for the variability of the enzyme properties among individuals and populations, genetic variants have not been investigated in CYP2D17. In the present study, CYP2D17 from 87 cynomolgus and 40 rhesus macaques was resequenced. The analysis found a total of 36 nonsynonymous variants, among which 5 were located in substrate recognition sites, the region important for protein function. Twenty-two variants were unique to cynomolgus macaques, of which 11 and 9 were found only in Indochinese and Indonesian cynomolgus macaques, respectively. Eight variants were unique to rhesus macaques. The functional characterization showed that two variant proteins (S188Y and V227I) heterologously expressed in Escherichia coli did not show substantial differences in the rate of bufuralol 1'-hydroxylation as compared with wild-type. However, measuring catalytic activities of the genotyped liver microsomes revealed that I297M and N337D were together significantly associated with higher rates, approximately 2.3- and 11.5-fold, of bufuralol 1'-hydroxylation and dextromethorphan O-demethylation, respectively, in the homozygotes than wild-type animals. The present study provided the first evidence that variability of a CYP2D-dependent metabolism in macaque liver is partly accounted for by CYP2D genotypes.

    DOI: 10.1124/dmd.114.059220

    PubMed

  • Uehara Shotaro, Murayama Norie, Nakanishi Yasuharu, Nakamura Chika, Hashizume Takanori, Zeldin Darryl C, Yamazaki Hiroshi, Uno Yasuhiro .  Immunochemical detection of cytochrome P450 enzymes in small intestine microsomes of male and female untreated juvenile cynomolgus monkeys. .  Xenobiotica44 ( 9 ) 769 - 774   2014年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The expression of small intestinal cytochromes P450 (P450s) has not been systematically measured in cynomolgus monkeys, which are widely used in preclinical drug studies to predict pharmacokinetics and toxicity in humans: therefore, P450 content of small intestine was quantified in 35 cynomolgus monkeys by immunoblotting using 11 selective antibodies. CYP2D, CYP2J2, CYP3A4 and CYP3A5 were detected in all 35 animals, while CYP1A and CYP2C9/19 were detected in 31 and 17 animals, respectively. CYP2C9 and CYP2C19 were detected with the same antibody. CYP1D, CYP2A, CYP2B6, CYP2C76 and CYP2E1 were not detected in any of the 35 animals examined. On analysis of pooled microsomes (35 animals), CYP3A (3A4+3A5) was most abundant (79% of total immunoquantified CYP1-3 proteins), followed by CYP2J2 (13%), CYP2C9/19 (4%), CYP1A (3%) and CYP2D (0.4%). On the analysis of individual microsome samples, each P450 content varied 2-to-6-fold between animals, and no sex differences were observed in any P450 content. These findings should help to increase the understanding of drug metabolism, especially the first-pass effect, in cynomolgus monkey small intestines.

    DOI: 10.3109/00498254.2014.895882

    PubMed

  • Yamazaki Miho, Shimizu Makiko, Uno Yasuhiro, Yamazaki Hiroshi .  Drug oxygenation activities mediated by liver microsomal flavin- containing monooxygenases 1 and 3 in humans, monkeys, rats, and minipigs. .  Biochem Pharmacol90 ( 2 ) 159 - 165   2014年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Liver microsomal flavin-containing monooxygenases (FMO, EC 1.14.13.8) 1 and 3 were functionally characterized in terms of expression levels and molecular catalytic capacities in human, cynomolgus monkey, rat, and minipig livers. Liver microsomal FMO3 in humans and monkeys and FMO1 and FMO3 in rats and minipigs could be determined immunochemically with commercially available anti-human FMO3 peptide antibodies or rat FMO1 peptide antibodies. With respect to FMO-dependent N-oxygenation of benzydamine and tozasertib and S-oxygenation of methimazole and sulindac sulfide activities, rat and minipig liver microsomes had high maximum velocity values (Vmax) and high catalytic efficiency (Vmax/Km, Michaelis constant) compared with those for human or monkey liver microsomes. Apparent Km values for recombinantly expressed rat FMO3-mediated N- and S-oxygenations were approximately 10-100-fold those of rat FMO1, although these enzymes had similar Vmax values. The mean catalytic efficiencies (Vmax/Km, 1.4 and 0.4 min(-1)muM(-1), respectively) of recombinant human and monkey FMO3 were higher than those of FMO1, whereas Vmax/Km values for rat and minipig FMO3 were low compared with those of FMO1. Minipig liver microsomal FMO1 efficiently catalyzed N- and S-oxygenation reactions; in addition, the minipig liver microsomal FMO1 concentration was higher than the levels in rats, humans, and monkeys. These results suggest that liver microsomal FMO1 could contribute to the relatively high FMO-mediated drug N- and S-oxygenation activities in rat and minipig liver microsomes and that lower expression of FMO1 in human and monkey livers could be a determinant factor for species differences in liver drug N- and S-oxygenation activities between experimental animals and humans.

    DOI: 10.1016/j.bcp.2014.04.019

    PubMed

  • Yajima Kanako, Uno Yasuhiro, Murayama Norie, Uehara Shotaro, Shimizu Makiko, Nakamura Chika, Iwasaki Kazuhide, Utoh Masahiro, Yamazaki Hiroshi .  Evaluation of 23 lots of commercially available cryopreserved hepatocytes for induction assays of human cytochromes P450. .  Drug Metab Dispos42 ( 5 ) 867 - 871   2014年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Due to the importance of in vitro cytochrome P450 (P450) induction assay to assess the possible drug-drug interaction events, the recent US Food and Drug Administration draft guidance and European Medicines Agency guideline recommend to assess P450 induction using fresh or cryopreserved hepatocytes at mRNA level and/or enzyme activity level. Although cryopreserved hepatocytes are commercially available for P450 induction assays, feasibility and practicability of these hepatocytes have not been fully investigated. In this study, a total of 23 lots of human cryopreserved hepatocytes were treated with three typical inducers (omeprazole, phenobarbital, and rifampicin), and induction of CYP1A2, CYP2B6, and CYP3A4 enzyme activity was measured. In 8 of these 23 hepatocyte lots, induction of CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP3A4 mRNA was also analyzed. The results revealed that CYP1A2, CYP2B6, and CYP3A4 were induced (>2.0-fold) by omeprazole, phenobarbital, and rifampicin, respectively, in all the hepatocyte lots tested at enzyme activity level (23 lots) and mRNA level (8 lots). In contrast, of the 8 hepatocyte lots treated with rifampicin, CYP2C8 and CYP2C9 mRNA were not induced in 5 and 2 hepatocyte lots, respectively, and CYP2C19 mRNA was not induced in any of the 8 hepatocyte lots tested. These results suggest that induction of CYP1A2, CYP2B6, and CYP3A4 can be readily assessed, but evaluation for CYP2C mRNA induction might not be feasible, using commercially available human cryopreserved hepatocytes.

    DOI: 10.1124/dmd.113.056804

    PubMed

  • Uno Yasuhiro, Utoh Masahiro, Iwasaki Kazuhide .  Polymorphisms of neonatal Fc receptor in cynomolgus and rhesus macaques. .  Drug Metab Pharmacokinet29 ( 5 ) 427 - 430   2014年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Neonatal Fc receptor (FcRn), a heterodimer of MHC class I-like protein and beta2-microglobulin, encoded by FCGRT and B2M, respectively, is important for recycling immunoglobulin G (IgG) antibodies by binding with the Fc region of IgG. Cynomolgus macaques are important animal species used in the evaluation of therapeutic antibodies, largely due to sequence similarities of target proteins to those of humans. Because the function of FcRn could be modified by mutations in FCGRT or B2M, 71 cynomolgus and 24 rhesus macaques were analyzed in the present study. A total of 21 variants were identified, of which 4 were non-synonymous in FCGRT. Fifteen variants were unique to cynomolgus macaques, of which 3, 2, and 5 were unique to cynomolgus macaques bred in China (MacfaCHN), Cambodia (MacfaCAM), and Indonesia (MacfaIDN), respectively. Five variants were shared by MacfaCHN and MacfaCAM, but not by MacfaIDN. In B2M, only 5 variants were found, including 2 non-synonymous variants. Tissue expression analysis showed that cynomolgus FCGRT and B2M were widely expressed in the 10 tissue types analyzed. None of the non-synonymous variants of FCGRT or B2M found changes in the amino acid residues known to be important for FcRn function, suggesting that substantial inter- animal variability of FcRn is not expected for the cynomolgus macaques analyzed.

    PubMed

  • Shimizu Makiko, Iwano Shunsuke, Uno Yasuhiro, Uehara Shotaro, Inoue Takashi, Murayama Norie, Onodera Jun, Sasaki Erika, Yamazaki Hiroshi .  Qualitative de novo analysis of full length cDNA and quantitative analysis of gene expression for common marmoset (Callithrix jacchus) transcriptomes using parallel long-read technology and short-read sequencing. .  PLoS One9 ( 6 ) e100936 - e100936   2014年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The common marmoset (Callithrix jacchus) is a non-human primate that could prove useful as human pharmacokinetic and biomedical research models. The cytochromes P450 (P450s) are a superfamily of enzymes that have critical roles in drug metabolism and disposition via monooxygenation of a broad range of xenobiotics; however, information on some marmoset P450s is currently limited. Therefore, identification and quantitative analysis of tissue-specific mRNA transcripts, including those of P450s and flavin-containing monooxygenases (FMO, another monooxygenase family), need to be carried out in detail before the marmoset can be used as an animal model in drug development. De novo assembly and expression analysis of marmoset transcripts were conducted with pooled liver, intestine, kidney, and brain samples from three male and three female marmosets. After unique sequences were automatically aligned by assembling software, the mean contig length was 718 bp (with a standard deviation of 457 bp) among a total of 47,883 transcripts. Approximately 30% of the total transcripts were matched to known marmoset sequences. Gene expression in 18 marmoset P450- and 4 FMO-like genes displayed some tissue-specific patterns. Of these, the three most highly expressed in marmoset liver were P450 2D-, 2E-, and 3A-like genes. In extrahepatic tissues, including brain, gene expressions of these monooxygenases were lower than those in liver, although P450 3A4 (previously P450 3A21) in intestine and P450 4A11- and FMO1-like genes in kidney were relatively highly expressed. By means of massive parallel long-read sequencing and short-read technology applied to marmoset liver, intestine, kidney, and brain, the combined next-generation sequencing analyses reported here were able to identify novel marmoset drug- metabolizing P450 transcripts that have until now been little reported. These results provide a foundation for mechanistic studies and pave the way for the use of marmosets as model animals for drug development in the future.

    DOI: 10.1371/journal.pone.0100936,

    PubMed

  • Utoh Masahiro, Murayama Norie, Uno Yasuhiro, Onose Yui, Hosaka Shinya, Fujino Hideki, Shimizu Makiko, Iwasaki Kazuhide, Yamazaki Hiroshi .  Monkey liver cytochrome P450 2C9 is involved in caffeine 7-N-demethylation to form theophylline. .  Xenobiotica43 ( 12 ) 1037 - 1042   2013年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Caffeine (1,3,7-trimethylxanthine) is a phenotyping substrate for human cytochrome P450 1A2. 3-N-Demethylation of caffeine is the main human metabolic pathway, whereas monkeys extensively mediate the 7-N-demethylation of caffeine to form pharmacological active theophylline. Roles of monkey P450 enzymes in theophylline formation from caffeine were investigated using individual monkey liver microsomes and 14 recombinantly expressed monkey P450 enzymes, and the results were compared with those for human P450 enzymes. Caffeine 7-N-demethylation activity in microsomes from 20 monkey livers was not strongly inhibited by alpha-naphthoflavone, quinidine or ketoconazole, and was roughly correlated with diclofenac 4'-hydroxylation activities. Monkey P450 2C9 had the highest activity for caffeine 7-N-demethylation. Kinetic analysis revealed that monkey P450 2C9 had a high Vmax/Km value for caffeine 7-N-demethylation, comparable to low Km value for monkey liver microsomes. Caffeine could dock favorably with monkey P450 2C9 modeled for 7-N-demethylation and with human P450 1A2 for 3-N-demethylation. The primary metabolite theophylline was oxidized to 8-hydroxytheophylline in similar ways by liver microsomes and by recombinant P450s in both humans and monkeys. These results collectively suggest a high activity for monkey liver P450 2C9 toward caffeine 7-N-demethylation, whereas, in humans, P450 1A2-mediated caffeine 3-N-demethylation is dominant.

    DOI: 10.3109/00498254.2013.793874

    PubMed

  • Takahashi Tsuyoshi, Ohtsuka Tatsuyuki, Yoshikawa Takahiro, Tatekawa Ichiro, Uno Yasuhiro, Utoh Masahiro, Yamazaki Hiroshi, Kume Toshiyuki .  Pitavastatin as an in vivo probe for studying hepatic organic anion transporting polypeptide-mediated drug-drug interactions in cynomolgus monkeys. .  Drug Metab Dispos41 ( 10 ) 1875 - 1882   2013年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Drug-drug interactions (DDIs) caused by the inhibition of hepatic uptake transporters such as organic anion transporting polypeptide (OATP) can affect therapeutic efficacy and cause adverse reactions. We investigated the potential utility of pitavastatin as an in vivo probe substrate for preclinically studying OATP-mediated DDIs using cynomolgus monkeys. Cyclosporine A (CsA) and rifampicin (RIF), typical OATP inhibitors, inhibited active uptake of pitavastatin into monkey hepatocytes with half-maximal inhibitory concentration values comparable with those in human hepatocytes. CsA and RIF increased the area under the plasma concentration-time curve (AUC) of intravenously administered pitavastatin in cynomolgus monkeys by 3.2- and 3.6-fold, respectively. In addition, there was no apparent prolongation of the elimination half-life of pitavastatin due to the decrease in both hepatic clearance and volume of distribution. These findings suggest that DDIs were caused by the inhibition of hepatic uptake of pitavastatin. CsA and RIF increased the AUC of orally administered pitavastatin by 10.6- and 14.8-fold, respectively, which was additionally caused by the effect of the CsA and RIF in the gastrointestinal tract. Hepatic contribution to the overall DDI for oral pitavastatin with CsA was calculated from the changes in hepatic availability and clearance, and it was shown that the magnitude of hepatic DDI was comparable between the present study and the clinical study. In conclusion, pharmacokinetic studies using pitavastatin as a probe in combination with drug candidates in cynomolgus monkeys are useful to support the assessment of potential clinical DDIs involving hepatic uptake transporters.

    DOI: 10.1124/dmd.113.052753

    PubMed

  • Uno Yasuhiro, Murayama Norie, Kunori Mutsuki, Yamazaki Hiroshi .  Systematic identification and characterization of glutathione S-transferases in cynomolgus macaque. .  Biochem Pharmacol86 ( 5 ) 679 - 690   2013年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Glutathione S-transferases (GSTs) are essential drug-metabolizing enzymes, involved in conjugation of various endogenous and exogenous substrates. Cynomolgus macaque is an important primate species in drug metabolism studies; however, cynomolgus GSTs have not been fully characterized. In this study the cDNAs of 12 GSTs (GSTA3-A5, GSTK1, GSTM2-M4, GSTO2, GSTP1, GSTS1, and GSTT1/2) were isolated from cynomolgus macaque and rhesus macaque liver. Cynomolgus GSTM1 cDNA was not amplified and only an aberrantly spliced GSTM1 transcript was isolated from rhesus macaque. Amino acid sequences of these 12 GSTs shared high sequence identities (93-98%) and were clustered into the same clades as the human orthologs in the phylogenetic tree. The 12 GSTs had exon-intron structures similar to the human orthologs, and exhibited distinct tissue expression patterns. GSTA3, GSTA5, and GSTM3/O2 were expressed predominantly in adrenal gland, jejunum, and testis, respectively, whereas the other GSTs showed universal expression patterns in the 10 tissues analyzed. Comparison of expression levels showed that GSTA1, GSTK1, GSTA3, and GSTM3 were most abundantly expressed in liver/jejunum, kidney, adrenal gland, and testis, respectively. Metabolic assays of proteins expressed heterologously in Escherichia coli, showed that all 12 GSTs and 5 previously identified GSTs, GSTA1/2, GSTM5, GSTO1, and GSTZ1, catalyzed the conjugation of GST substrate(s) 1-chloro-2,4-dinitrobenzene and/or 1,2-epoxy-3-(p-nitrophenoxy)propane, indicating that these 17 GSTs are functional drug-metabolizing enzymes. These results suggest that the 12 GST genes examined in this study are expressed and encoded functional enzymes in cynomolgus macaque.

    DOI: 10.1016/j.bcp.2013.06.022

    PubMed

  • Uno Yasuhiro, Murayama Norie, Kunori Mutsuki, Yamazaki Hiroshi .  Characterization of microsomal glutathione S-transferases MGST1, MGST2, and MGST3 in cynomolgus macaque. .  Drug Metab Dispos41 ( 9 ) 1621 - 1625   2013年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The glutathione S-transferase (GST) family comprises cytosolic, mitochondrial, and microsomal GSTs, all essential enzymes that metabolize a wide range of endogenous and exogenous substrates. Among the microsomal GSTs (MGSTs) in humans, MGST1, MGST2, and MGST3 are involved in detoxification; however, MGSTs have not been fully investigated in cynomolgus macaque, an important primate species widely used in drug metabolism and toxicity studies. In the present study, cynomolgus MGST2 and MGST3 cDNAs were isolated from liver tissue and characterized along with previously isolated cynomolgus MGST1. For comparison with the cynomolgus cDNAs, MGST2 and MGST3 cDNAs were also isolated from rhesus macaque (closely related to cynomolgus macaque) liver. Cynomolgus MGST2 and MGST3, respectively, were highly identical (99 and 98%) to human MGST2 and MGST3 and nearly identical to the amino acid sequences of the rhesus orthologs, and they were closely clustered with human MGST2 and MGST3 by phylogenetic analysis. The analysis of genome data indicated that MGST1, MGST2, and MGST3, respectively, had similar gene structures and genomic organization in macaque and human. Therefore, cynomolgus MGSTs have molecular similarities to the corresponding human MGSTs. Cynomolgus MGST2 and MGST3 were expressed in liver, jejunum, and kidney, but at lower levels than MGST1. GST activities were measured with 1-chloro-2,4-dinitrobenzene and 1,2-epoxy-3-(p-nitrophenoxy)propane as substrates, using proteins heterologously expressed in Escherichia coli. Cynomolgus MGST1, MGST2, and MGST3 conjugated 1-chloro-2,4-dinitrobenzene and 1,2-epoxy-3-(p-nitrophenoxy)propane, indicating that cynomolgus MGST1, MGST2, and MGST3 are functional enzymes. These results suggest that these functional cynomolgus MGST enzymes and the corresponding human MGSTs are molecularly similar.

    DOI: 10.1124/dmd.113.052977

    PubMed

  • Uno Yasuhiro, Shimizu Makiko, Yamazaki Hiroshi .  Molecular and functional characterization of flavin-containing monooxygenases in cynomolgus macaque. .  Biochem Pharmacol85 ( 12 ) 1837 - 1847   2013年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Flavin-containing monooxygenases (FMOs), drug-metabolizing enzymes essential for the metabolism of endogenous biochemicals and foreign compounds, have been characterized in human (including FMO1-5 and FMO6P), but remain to be investigated in cynomolgus macaque. In this study, cDNAs of cynomolgus FMO1-5 and FMO6 were isolated and characterized. Amino acid sequences of cynomolgus FMO1-5, respectively, shared high sequence identities (94-98%) and were closely clustered in a phylogenetic tree, with human FMO1-5. Eight different transcripts, due to alternative splicing, were isolated for cynomolgus FMO6, which is highly identical (~96%) to human FMO6P. Among the 10 tissue types analyzed, cynomolgus FMO1, FMO2, FMO4, and FMO6 were most abundantly expressed in kidney, while cynomolgus FMO3 and FMO5 were most abundantly expressed in liver. In kidney and liver, the most abundantly expressed cynomolgus FMO genes were FMO1 and FMO3, respectively. Cynomolgus FMO1, FMO2, FMO3, and FMO5 metabolized benzydamine, and FMO1/FMO3 and FMO3 also metabolized methimazole and trimethylamine, respectively. Rates of benzydamine N-oxygenation (catalyzed by FMO3) varied (approximately 20-fold) among the 28 cynomolgus livers and were significantly correlated with FMO3 protein expression, indicating that the inter-animal variations in benzydamine N-oxygenation might be partly accounted for by the variable FMO3 expression. Cynomolgus FMO6 metabolized benzydamine only slightly, but minimal expression of FMO6 in all tissue precludes the importance of FMO6 in drug metabolism, unlike cynomolgus FMO1, FMO2, FMO3, and FMO5 which were all functional. Abundant expression of FMO1 and FMO3 in kidney and liver, respectively, suggest their importance in drug metabolism in cynomolgus macaque, similar to human.

    DOI: 10.1016/j.bcp.2013.04.012

    PubMed

  • Emoto Chie, Yoda Noriaki, Uno Yasuhiro, Iwasaki Kazuhide, Umehara Ken, Kashiyama Eiji, Yamazaki Hiroshi .  Comparison of p450 enzymes between cynomolgus monkeys and humans: p450 identities, protein contents, kinetic parameters, and potential for inhibitory profiles. .  Curr Drug Metab14 ( 2 ) 239 - 252   2013年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys are used to predict human pharmacokinetic and/or toxic profiles in the drug developmental stage. Cynomolgus P450s exhibit a high degree of identity (more than 90%) in both cDNA and amino acid sequences with corresponding human P450s. CYP3A protein predominantly exists in cynomolgus monkey liver microsomes, followed by CYP2A, CYP2C, CYP2B6, CYP2E1, and CYP2D. There are many similarities of metabolic properties in cytochrome P450s between cynomolgus monkeys and humans, but the species differences between cynomolgus monkey and human P450s are clearly present in substrate specificity and inhibitor selectivity. Diclofenac 4'-hydroxylation (DFOH) in monkey liver and intestinal microsomes shows much lower activities compared with those in human liver and intestinal microsomes. Sulfaphenazole strongly inhibits DFOH in human liver microsomes, but does not effectively inhibit DFOH in monkey liver and intestinal microsomes. Cynomolgus CYP2C19 exhibits higher activity for DFOH than cynomolgus CYP2C9 although this reaction is a marker reaction of human CYP2C9. On the other hand, cynomolgus CYP2C76 orthologue is not expressed in humans and shows 70-72% identity in amino acid sequences of human CYP2C subfamilies. Cynomolgus CYP2C76 metabolizes non-CYP2C substrates, 7-ethoxyresorufin (human CYP1A substrate) and bufuralol (human CYP2D6 substrate). In addition, cynomolgus CYP3A4 and CYP3A5 also exhibits wider substrate selectivity toward human CYP2D6 and CYP2E1 substrates. These enzymes may be responsible for species difference in drug metabolism between cynomolgus monkeys and humans. The comparative data presented here can be helpful for designing in vivo metabolic assays using cynomolgus monkeys in terms of substrate specificity and inhibitor selectivity.

    PubMed

  • Nakanishi Yasuharu, Yamashita Hiroyuki, Yoshikawa Tsuyoshi, Tominaga Takeshi, Nojiri Koichiro, Sunaga Yoshiharu, Muneoka Atsunobu, Iwasaki Kazuhide, Utoh Masahiro, Nakamura Chika, Yamazaki Hiroshi, Uno Yasuhiro .  Cytochrome P450 metabolic activities in the small intestine of cynomolgus macaques bred in Cambodia, China, and Indonesia. .  Drug Metab Pharmacokinet28 ( 6 ) 510 - 513   2013年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus macaques, used in drug metabolism studies due to their evolutionary closeness to humans, are mainly bred in Asian countries, including Cambodia, China, and Indonesia. Cytochromes P450 (P450s) are important drug-metabolizing enzymes, present in the liver and small intestine, major drug metabolizing organs. Previously, our investigation did not find statistically significant differences in hepatic P450 metabolic activities measured in cynomolgus macaques bred in Cambodia (MacfaCAM) and China (MacfaCHN). In the present study, P450 metabolic activity was investigated in the small intestine of MacfaCAM and MacfaCHN, and cynomolgus macaques bred in Indonesia (MacfaIDN) using P450 substrates, including 7-ethoxyresorufin, coumarin, bupropion, paclitaxel, diclofenac, S-mephenytoin, bufuralol, chlorzoxazone, and testosterone. The results indicated that P450 metabolic activity of the small intestine was not statistically significantly different (<2.0-fold) in MacfaCAM, MacfaCHN, and MacfaIDN. In addition, statistically significant sex differences were not observed (<2.0-fold) in any P450 metabolic activity in MacfaCAM as supported by mRNA expression results. These results suggest that P450 metabolic activity of the small intestine does not significantly differ statistically among MacfaCAM, MacfaCHN, and MacfaIDN.

    PubMed

  • Hosoi Yoshio, Uno Yasuhiro, Murayama Norie, Fujino Hideki, Shukuya Mitsunori, Iwasaki Kazuhide, Shimizu Makiko, Utoh Masahiro, Yamazaki Hiroshi .  Monkey liver cytochrome P450 2C19 is involved in R- and S-warfarin 7-hydroxylation. .  Biochem Pharmacol84 ( 12 ) 1691 - 1695   2012年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkeys are widely used as primate models in preclinical studies. However, some differences are occasionally seen between monkeys and humans in the activities of cytochrome P450 enzymes. R- and S-warfarin are model substrates for stereoselective oxidation in humans. In this current research, the activities of monkey liver microsomes and 14 recombinantly expressed monkey cytochrome P450 enzymes were analyzed with respect to R- and S-warfarin 6- and 7-hydroxylation. Monkey liver microsomes efficiently mediated both R- and S-warfarin 7-hydroxylation, in contrast to human liver microsomes, which preferentially catalyzed S-warfarin 7-hydroxylation. R-Warfarin 7-hydroxylation activities in monkey liver microsomes were not inhibited by alpha-naphthoflavone or ketoconazole, and were roughly correlated with P450 2C19 levels and flurbiprofen 4-hydroxylation activities in microsomes from 20 monkey livers. In contrast, S-warfarin 7-hydroxylation activities were not correlated with the four marker drug oxidation activities used. Among the 14 recombinantly expressed monkey P450 enzymes tested, P450 2C19 had the highest activities for R- and S-warfarin 7-hydroxylations. Monkey P450 3A4 and 3A5 slowly mediated R- and S-warfarin 6-hydroxylations. Kinetic analysis revealed that monkey P450 2C19 had high V(max) and low K(m) values for R-warfarin 7-hydroxylation, comparable to those for monkey liver microsomes. Monkey P450 2C19 also mediated S-warfarin 7-hydroxylation with V(max) and V(max)/K(m) values comparable to those for recombinant human P450 2C9. R-warfarin could dock favorably into monkey P450 2C19 modeled. These results collectively suggest high activities for monkey liver P450 2C19 toward R- and S-warfarin 6- and 7-hydroxylation in contrast to the saturation kinetics of human P450 2C9-mediated S-warfarin 7-hydroxylation.

    DOI: 10.1016/j.bcp.2012.09.026

    PubMed

  • Kuribayashi Shunji, Uno Yasuhiro, Naito Shinsaku, Yamazaki Hiroshi .  Different metabolites of human hepatotoxic pyrazolopyrimidine derivative 5-n-butyl-pyrazolo[1,5-a]pyrimidine produced by human, rat and monkey cytochrome P450 1A2 and liver microsomes. .  Basic Clin Pharmacol Toxicol110 ( 4 ) 405 - 408   2012年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1111/j.1742-7843.2011.00823.x

    PubMed

  • Uehara Shotaro, Murayama Norie, Yamazaki Hiroshi, Uno Yasuhiro .  CYP2C76 non-synonymous variants in cynomolgus and rhesus macaques. .  Drug Metab Pharmacokinet27 ( 3 ) 344 - 348   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus CYP2C76, not orthologous to any human cytochrome P450, partly accounts for species differences in drug metabolism between cynomolgus macaques and humans. To discover the CYP2C76 variants, we previously surveyed cynomolgus macaque genomes and found several non-synonymous variants, including a null allele. However, the analysis was limited to cynomolgus macaques, and the number of genomes was relatively small. In this study, therefore, further screening was conducted using 74 cynomolgus and 30 rhesus macaques. A total of 18 non-synonymous variants was found, among which 7 were in substrate recognition sites, important for protein function, and 14 (74%) were shared by both macaque lineages. In cynomolgus macaques, 3 (16%) non-synonymous variants were unique to Indochinese animals, whereas all the variants found in Indonesian animals were shared by Indochinese animals. Among the 18 variants, as compared with the wild type, in progesterone 16alpha-hydroxylation, L65F, M310L, and N364S variants showed lower metabolic activity and lower intrinsic clearance by kinetic analysis. Molecular modeling indicated that the reduced catalytic activity of the L65F variant in progesterone 16alpha- hydroxylation possibly resulted from a longer distance of progesterone to the heme in the active site of the CYP2C76 protein. L65F, M310L, and N364S variants might partly influence inter-animal variations of CYP2C76 metabolic activities.

    PubMed

  • Ise Ryota, Kito Go, Uno Yasuhiro .  Expression profile of early estradiol-responsive genes in cynomolgus macaque liver: implications for drug-metabolizing enzymes. .  Drug Metab Pharmacokinet27 ( 4 ) 451 - 455   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Estrogen plays important roles in estrogen-responsive tissues, such as mammary glands, ovaries, and the uterus. In the liver, the major drug metabolizing organ, estrogen is known to regulate expression of some drug-metabolizing enzymes. Due to the lack of information on the role of estrogen in hepatic gene expression in primate species, we previously investigated the late response of hepatic gene expression to estradiol in cynomolgus macaques. To understand the early response of hepatic gene expression to estradiol, in this study, microarray analysis was conducted using cynomolgus macaque liver samples collected at 1 h and 5 h after estradiol injection. Comparison of expression profiles in estradiol and solvent (control)-treated ovariectomized cynomolgus macaques revealed 27 differentially expressed genes (>2.0-fold), including 18 at 1 h and 9 at 5 h after estradiol injection. As indicated by Gene Ontology analysis, these genes were related to oxidoreductase activity and transferase activity, partly representing important aspects of drug-metabolizing enzymes. Further analysis by quantitative polymerase chain reaction revealed that estradiol down-regulated CYP2A24, CYP2C76, and CYP2E1 (>2.0-fold) at 1 h and up-regulated GSTM5 (>2.0-fold) at 5 h after estradiol injection. These results suggest that the short-term estradiol treatment influenced expression of hepatic genes, including drug- metabolizing enzyme genes, in cynomolgus macaque liver.

    PubMed

  • Ise Ryota, Nakanishi Yasuharu, Kohara Sakae, Yamashita Hiroyuki, Yoshikawa Tsuyoshi, Iwasaki Kazuhide, Nagata Ryoichi, Fukuzaki Koichiro, Utoh Masahiro, Nakamura Chika, Yamazaki Hiroshi, Uno Yasuhiro .  Expression profile of hepatic genes in cynomolgus macaques bred in Cambodia, China, and Indonesia: implications for cytochrome P450 genes. .  Drug Metab Pharmacokinet27 ( 3 ) 307 - 316   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus macaques, frequently used in drug metabolism studies, are bred mainly in the countries of Asia; however, comparative studies of drug metabolism between cynomolgus macaques bred in these countries have not been conducted. In this study, hepatic gene expression profiles of cynomolgus macaques bred in Cambodia (mfCAM), China (mfCHN), and Indonesia (mfIDN) were analyzed. Microarray analysis revealed that expression of most hepatic genes, including drug-metabolizing enzyme genes, was not substantially different between mfCAM, mfCHN, and mfIDN; only 1.1% and 3.0% of all the gene probes detected differential expression (>2.5-fold) in mfCAM compared with mfCHN and mfIDN, respectively. Quantitative polymerase chain reaction showed that the expression levels of 14 cytochromes P450 (P450s) important for drug metabolism did not differ (>2.5-fold) in mfCAM, mfCHN, and mfIDN, validating the microarray data. In contrast, expression of CYP2B6 and CYP3A4 differed (>2.5-fold, p < 0.05) between cynomolgus (mfCAM, mfCHN, or mfIDN) and rhesus macaques, indicating greater differences in expression of P450 genes between the two lineages. Moreover, metabolic activities measured using 14 P450 substrates did not differ substantially (<1.5-fold) between mfCAM and mfCHN. These results suggest that gene expression profiles, including drug-metabolizing enzyme genes such as P450 genes, are similar in mfCAM, mfCHN, and mfIDN.

    PubMed

  • Honda Kouichi, Komatsu Tetsuya, Koyama Fumika, Kubota Akira, Kawakami Kei, Asakura Hiroyuki, Uno Yasuhiro, Kitazawa Takio, Hiraga Takeo, Teraoka Hiroki .  Expression of two novel cytochrome P450 3A131 and 3A132 in liver and small intestine of domestic cats. .  J Vet Med Sci73 ( 11 ) 1489 - 1492   2011年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cytochrome P450 3A (CYP3A) is the major subfamily of CYP, one of the most important metabolizing enzymes for drugs in humans and other mammals. We found two novel CYP3A genes, CYP3A131 and CYP3A132 in domestic cats (Felis catus). Both feline CYP3A proteins consist of 504 deduced amino acids and show high identity with canine CYP3A homologues and those of some artiodactyls. CYP3A131 transcripts were expressed predominantly in liver and small intestine, and to a negligible extent in other tissues, including brain, heart, kidney and lung. CYP3A132 expression was only detected in liver with much lesser amount. These results suggest the possible major role of CYP3A131 in xenobiotic metabolism including first- pass effects in domestic cats.

    DOI: 10.1292/jvms.11-0098

    PubMed

  • Uno Yasuhiro, Kito Go .  Effect of estradiol on gene expression profile in cynomolgus macaque liver: implications for drug-metabolizing enzymes. .  Drug Metab Dispos39 ( 11 ) 2003 - 2007   2011年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Estrogen regulation of gene expression is essential for physiological function of estrogen-responsive tissues, such as mammary glands, ovaries, and the uterus. In the liver, estrogen is responsible for sex-dependent gene expression of drug-metabolizing enzymes in rodents. However, the influence of estrogen on hepatic gene expression has not been fully investigated in primates, including human. Macaque, including cynomolgus macaque, is an important species for comparative studies aimed at understanding human physiology due to its evolutionary closeness to human. To identify estrogen-responsive genes in primate liver, therefore, hepatic gene expression was compared, by microarray analysis, in ovariectomized cynomolgus macaques treated with estradiol or solvent (control). The analysis identified 98 estradiol-responsive genes; 47 and 51 were up- and down-regulated by estradiol, respectively (>/=2.0-fold, P < 0.05). Expression of drug-metabolizing enzyme genes was also influenced by estradiol treatment; estradiol enhanced expression of GSTM5 (3.8-fold, P < 0.05) and CYP3A8(4) (2.7-fold, P < 0.01), but lowered expression of CYP4F12 (2.2-fold, P < 0.01), as verified by quantitative polymerase chain reaction. In particular, CYP3A8(4), orthologous to human CYP3A4, is an essential drug-metabolizing enzyme in cynomolgus macaque liver. These results suggest that expression of hepatic genes, including drug- metabolizing enzyme genes, is at least partly regulated by estradiol in cynomolgus macaque.

    DOI: 10.1124/dmd.111.041004

    PubMed

  • Uehara Shotaro, Murayama Norie, Nakanishi Yasuharu, Zeldin Darryl C, Yamazaki Hiroshi, Uno Yasuhiro .  Immunochemical detection of cytochrome P450 enzymes in liver microsomes of 27 cynomolgus monkeys. .  J Pharmacol Exp Ther339 ( 2 ) 654 - 661   2011年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The cynomolgus monkey is widely used as a primate model in preclinical studies because of its evolutionary closeness to humans. Despite their importance in drug metabolism, the content of each cytochrome P450 (P450) enzyme has not been systematically determined in cynomolgus monkey livers. In this study, liver microsomes of 27 cynomolgus monkeys were analyzed by immunoblotting using selective P450 antibodies. The specificity of each antibody was confirmed by analyzing the cross- reactivity against 19 CYP1-3 subfamily enzymes using recombinant proteins. CYP2A, CYP2B6, CYP2C9/19, CYP2C76, CYP2D, CYP2E, CYP3A4, and CYP3A5 were detected in all 27 animals. In contrast, CYP1A, CYP1D, and CYP2J were below detectable levels in all liver samples. The average content of each P450 showed that among the P450s analyzed CYP3A (3A4 and 3A5) was the most abundant (40% of total immunoquantified P450), followed by CYP2A (25%), CYP2C (14%), CYP2B6 (13%), CYP2E1 (11%), and CYP2D (3%). No apparent sex differences were found for any P450. Interanimal variations ranged from 2.6-fold (CYP3A) to 11-fold (CYP2C9/19), and most P450s (CYP2A, CYP2D, CYP2E, CYP3A4, and CYP3A5) varied 3- to 4-fold. To examine the correlations of P450 content with enzyme activities, metabolic assays were performed in 27 cynomolgus monkey livers using 7-ethoxyresorufin, coumarin, pentoxyresorufin, flurbiprofen, bufuralol, dextromethorphan, and midazolam. CYP2D and CYP3A4 contents were significantly correlated with typical reactions of human CYP2D (bufuralol 1'-hydroxylation and dextromethorphan O-deethylation) and CYP3A (midazolam 1'-hydroxylation and 4-hydroxylation). The results presented in this study provide useful information for drug metabolism studies using cynomolgus monkeys.

    DOI: 10.1124/jpet.111.185009

    PubMed

  • Uno Yasuhiro, Osada Naoki .  CpG site degeneration triggered by the loss of functional constraint created a highly polymorphic macaque drug-metabolizing gene, CYP1A2. .  BMC Evol Biol11   283 - 283   2011年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Elucidating the pattern of evolutionary changes in drug- metabolizing genes is an important subject not only for evolutionary but for biomedical research. We investigated the pattern of divergence and polymorphisms of macaque CYP1A1 and CYP1A2 genes, which are major drug- metabolizing genes in humans. In humans, CYP1A2 is specifically expressed in livers while CYP1A1 has a wider gene expression pattern in extrahepatic tissues. In contrast, macaque CYP1A2 is expressed at a much lower level than CYP1A1 in livers. Interestingly, a previous study has shown that Macaca fascicularis CYP1A2 harbored unusually high genetic diversity within species. Genomic regions showing high genetic diversity within species is occasionally interpreted as a result of balancing selection, where natural selection maintains highly diverged alleles with different functions. Nevertheless many other forces could create such signatures. RESULTS: We found that the CYP1A1/2 gene copy number and orientation has been highly conserved among mammalian genomes. The signature of gene conversion between CYP1A1 and CYP1A2 was detected, but the last gene conversion event in the simian primate lineage occurred before the Catarrhini-Platyrrhini divergence. The high genetic diversity of macaque CYP1A2 therefore cannot be explained by gene conversion between CYP1A1 and CYP1A2. By surveying CYP1A2 polymorphisms in total 91 M. fascicularis and M. mulatta, we found several null alleles segregating in these species, indicating functional constraint on CYP1A2 in macaques may have weakened after the divergence between humans and macaques. We propose that the high genetic diversity in macaque CYP1A2 is partly due to the degeneration of CpG sites, which had been maintained at a high level by purifying selection, and the rapid degeneration process was initiated by the loss of functional constraint on macaque CYP1A2. CONCLUSIONS: Our findings show that the highly polymorphic CYP1A2 gene in macaques has not been created by balancing selection but by the burst of CpG site degeneration after loss of functional constraint. Because the functional importance of CYP1A1/2 genes is different between humans and macaques, we have to be cautious in extrapolating a drug-testing data using substrates metabolized by CYP1A genes from macaques to humans, despite of their somewhat overlapping substrate specificity.

    DOI: 10.1186/1471-2148-11-283

    PubMed

  • Uno Yasuhiro, Matsuno Kiyomi, Murayama Norie, Nakamura Chika, Yamazaki Hiroshi .  Metabolism of P450 probe substrates by cynomolgus monkey CYP2C76. .  Basic Clin Pharmacol Toxicol109 ( 4 ) 315 - 318   2011年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1111/j.1742-7843.2011.00740.x

    PubMed

  • Uno Yasuhiro, Matsushita Akinori, Yamazaki Hiroshi .  CYP1B1 is polymorphic in cynomolgus and rhesus macaques. .  J Vet Med Sci73 ( 9 ) 1229 - 1231   2011年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cytochrome P450 (CYP) 1B1 is involved in the metabolic activation of various procarcinogens, and some CYP1B1 genetic variants alter CYP1B1-dependent procarcinogen metabolism. Cynomolgus and rhesus macaques are frequently used in toxicity tests due to their evolutionary closeness to humans. In this study, we attempted to identify CYP1B1 genetic variants in 13 cynomolgus and 4 rhesus macaques. A total of 17 genetic variants were identified, including 8 non-synonymous genetic variants, indicating that, similar to humans, CYP1B1 is polymorphic in macaques. These CYP1B1 genetic variants could be the basis for understanding potential inter-animal differences in macaque CYP1B1-dependent metabolism of promutagens.

    DOI: 10.1292/jvms.11-0137

    PubMed

  • Uno Yasuhiro, Iwasaki Kazuhide, Yamazaki Hiroshi, Nelson David R .  Macaque cytochromes P450: nomenclature, transcript, gene, genomic structure, and function. .  Drug Metab Rev43 ( 3 ) 346 - 361   2011年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Monkeys, especially macaques, including cynomolgus (Macaca fascicularis) and rhesus monkeys (Macaca mulatta), are frequently used in drug metabolism studies due to their evolutionary closeness to humans. Recently, numerous cytochrome P450 (P450 or CYP) cDNAs have been identified and characterized in cynomolgus and rhesus monkeys and were named by the P450 Nomenclature Committee. However, recent advances in genome analysis of cynomolgus and rhesus monkeys revealed that some monkey P450s are apparently orthologous to human P450s and thus need to be renamed corresponding to their human orthologs. In this review, we focus on the P450s identified in cynomolgus and rhesus monkeys and present an overview of the identity and functional characteristics of each P450 cDNA in the CYP1-4 families. Information on the Japanese monkey (Macaca fuscata), African green monkey (Cercopithecus aethiops), and marmoset (Callithrix jacchus), primate species used in some drug metabolism studies, are also included. We compared the genomic structure of the macaque P450 genes to those of human and rat P450 genes in the CYP1-4 families. Based on sequence identity, phylogeny, and genomic organization of monkey P450s, we determined orthologous relationships of monkey P450s and, in this article, propose a revised nomenclature: CYP2B17/CYP2B30 to CYP2B6, CYP2C20/CYP2C74 to CYP2C8, CYP2C43/CYP2C83 to CYP2C9, CYP2C75 to CYP2C19, CYP2F6 to CYP2F1, CYP3A8/CYP3A21/CYP3A64 to CYP3A4, CYP3A66 to CYP3A5, and CYP4F45 to CYP4F2. The information presented in this review is expected to promote a better understanding of monkey P450 genes through comparative genomics and thereby make it more feasible to use monkeys in drug metabolism studies.

    DOI: 10.3109/03602532.2010.549492

    PubMed

  • Ise Ryota, Uehara Shotaro, Akiyama Hideo, Kondo Satoshi, Iwasaki Kazuhide, Nagata Ryoichi, Nobumasa Hitoshi, Yamazaki Hiroshi, Uno Yasuhiro .  A newly developed DNA microarray is useful to assess induction of cytochromes p450 in the cynomolgus monkey. .  Drug Metab Pharmacokinet26 ( 3 ) 228 - 235   2011年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cytochromes P450 (P450s or CYPs) are a gene family of highly homologous genes and include the CYP1-4 family, which is relevant to drug metabolism. In the cynomolgus monkey (which is frequently used in drug metabolism studies), numerous CYPs (mfCYPs) have been identified in the CYP1-4 family. DNA microarrays are useful for high-throughput screening assays; however, there is a potential problem with cross-hybridization of highly homologous genes in the gene family. This problem might be solved with the use of low-density DNA microarrays, with which specific validation can be performed for the genes on the microarray. We have developed a DNA microarray for the 20 mfCYPs and have evaluated and validated its specificity and usefulness. First, in both DNA microarray and quantitative polymerase chain reaction (qPCR) analyses, hepatic expression of each mfCYP correlated well, and similar tissue expression patterns were observed for five representative mfCYPs, confirming the specificity of the DNA microarray. Second, the usefulness of this DNA microarray was validated by induction analysis of mfCYPs in primary hepatocytes, which successfully detected known responders, but also novel responders (mfCYP2C43, mfCYP2C75, and mfCYP3A5 for rifampicin), as confirmed by qPCR analysis. This DNA microarray can thus be utilized for high-throughput assays during drug development.

    DOI: 10.2133/dmpk.DMPK-10-RG-099

    PubMed

  • Nakanishi Yasuharu, Matsushita Akinori, Matsuno Kiyomi, Iwasaki Kazuhide, Utoh Masahiro, Nakamura Chika, Uno Yasuhiro .  Regional distribution of drug-metabolizing enzyme activities in the liver and small intestine of cynomolgus monkeys. .  Drug Metab Pharmacokinet26 ( 3 ) 288 - 294   2011年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The cynomolgus monkey is an animal species widely used to study drug metabolism because of its evolutionary closeness to humans. However, drug-metabolizing enzyme activities have not been compared in various parts of the liver and small intestine in cynomolgus monkeys. In this study, therefore, drug-metabolizing enzyme activities were analyzed in the liver (the five lobes) and small intestine (six sections from the duodenum to the distal ileum). 7-Ethoxyresorufin O-deethylation, coumarin 7-hydroxylation, paclitaxel 6alpha-hydroxylation, diclofenac 4'-hydroxylation, tolbutamide methylhydroxylation, S-mephenytoin 4'-hydroxylation, bufuralol 1'-hydroxylation, chlorzoxazone 6-hydroxylation, midazolam 1'-hydroxylation, and testosterone 6beta-, 16alpha-, 16beta-, and 2alpha-hydroxylation were used as the probe reactions for this investigation. In liver, all probe reactions were detected and enzyme activity levels were similar in all lobes, whereas, in the small intestine, all enzyme activities were detected (except for coumarin 7-hydroxylase and testosterone 16alpha-hydroxylase activity), but from jejunum to ileum there was a decrease in the level of enzyme activity. This includes midazolam 1'-hydroxylation and testosterone 6beta-hydroxylation, which are catalyzed by cynomolgus monkey cytochrome P450 (CYP) 3A4/5, orthologs of human CYP3A4/5, which are important drug- metabolizing enzymes. The data presented in this study are expected to facilitate the use of cynomolgus monkeys in drug metabolism studies.

    DOI: 10.2133/dmpk.DMPK-10-NT-101

    PubMed

  • Uno Yasuhiro, Matsuno Kiyomi, Nakamura Chika, Utoh Masahiro, Yamazaki Hiroshi .  Cynomolgus macaque CYP4 isoforms are functional, metabolizing arachidonic acid. .  J Vet Med Sci73 ( 4 ) 487 - 490   2011年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cytochrome P450 (CYP) is important for metabolism of not only xenobiotics such as drugs, but also endogenous compounds including arachidonic acids. CYP4A11, CYP4F3v2, CYP4F11, and CYP4F45 have been identified in cynomolgus macaque, an animal species widely used for investigation of drug metabolism due to its evolutionary closeness to human. However, their metabolic functions have not been investigated. In this study, proteins were heterologously expressed in Escherichia coli and characterized by metabolic assays using arachidonic acids as substrates that are metabolized by CYP4 isoforms in human. The results showed that all four CYPs metabolized arachidonic acids. Therefore, cynomolgus macaque CYP4A11, CYP4F3v2, CYP4F11, and CYP4F45 are functional enzymes.

    DOI: 10.1292/jvms.10-0333

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Murayama Norie, Yamazaki Hiroshi .  CYP2G2, pseudogenized in human, is expressed in nasal mucosa of cynomolgus monkey and encodes a functional drug-metabolizing enzyme. .  Drug Metab Dispos39 ( 4 ) 717 - 723   2011年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    CYP2G2P is pseudogenized in humans because of two nonsense mutations (c.76C>T in exon 1 and c.382C>T in exon 3) in the putative coding region of the gene sequence, whereas mouse, rat, and rabbit CYP2Gs are expressed and functional in nasal mucosa. In this study, we assessed the intactness of CYP2G in a cynomolgus monkey, a macaque species important for drug metabolism studies because of its evolutionary closeness to human. On the basis of a gene sequence (highly identical to human CYP2G2P) found in the macaque genome, CYP2G2 cDNA was successfully isolated from cynomolgus monkey nasal mucosa. CYP2G2 cDNA, containing an open reading frame of 494 amino acids, was shown to share high sequence identity (nearly 95%) with the putative coding region of human CYP2G2P. Cynomolgus monkey CYP2G2 shared the highest sequence identity (59-61%) with CYP2A23, CYP2A24, and CYP2A26 among cynomolgus monkey cytochromes P450. Cynomolgus monkey CYP2G2 mRNA was predominantly expressed in the nasal mucosa, where CYP2G2 protein expression was also detected. Metabolic assays indicated that cynomolgus monkey CYP2G2 metabolized coumarin, similar to cynomolgus monkey CYP2A23, CYP2A24, and CYP2A26. Moreover, among 39 cynomolgus monkeys and 11 rhesus monkeys examined in this study, only 2 cynomolgus monkeys and 1 rhesus monkey were heterozygous for c.76C>T. No animals carried c.382C>T. These results suggest that cynomolgus monkey CYP2G2 is a functional drug-metabolizing enzyme in nasal mucosa.

    DOI: 10.1124/dmd.110.036574

    PubMed

  • Chie Emoto, Kazuhide Iwasaki, Ryo Koizumi, Masahiro Utoh, Norie Murayama, Yasuhiro Uno, Hiroshi Yamazaki .  Species difference between cynomolgus monkeys and humans on cytochromes P450 2D and 3A-dependent drug oxidation activities in liver microsomes .  J Health Sci 57 ( 2 ) 164 - 170   2011年4月査読

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Uno Yasuhiro, Uehara Shotaro, Kohara Sakae, Iwasaki Kazuhide, Nagata Ryoichi, Fukuzaki Koichiro, Utoh Masahiro, Murayama Norie, Yamazaki Hiroshi .  Newly identified CYP2C93 is a functional enzyme in rhesus monkey, but not in cynomolgus monkey. .  PLoS One6 ( 2 ) e16923 - e16923   2011年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkey and rhesus monkey are used in drug metabolism studies due to their evolutionary closeness and physiological resemblance to human. In cynomolgus monkey, we previously identified cytochrome P450 (P450 or CYP) 2C76 that does not have a human ortholog and is partly responsible for species differences in drug metabolism between cynomolgus monkey and human. In this study, we report characterization of CYP2C93 cDNA newly identified in cynomolgus monkey and rhesus monkey. The CYP2C93 cDNA contained an open reading frame of 490 amino acids approximately 84-86% identical to human CYP2Cs. CYP2C93 was located in the genomic region, which corresponded to the intergenic region in the human genome, indicating that CYP2C93 does not correspond to any human genes. CYP2C93 mRNA was expressed predominantly in the liver among 10 tissues analyzed. The CYP2C93 proteins heterologously expressed in Escherichia coli metabolized human CYP2C substrates, diclofenac, flurbiprofen, paclitaxel, S-mephenytoin, and tolbutamide. In addition to a normal transcript (SV1), an aberrantly spliced transcript (SV2) lacking exon 2 was identified, which did not give rise to a functional protein due to frameshift and a premature termination codon. Mini gene assay revealed that the genetic variant IVS2-1G>T at the splice site of intron 1, at least partly, accounted for the exon-2 skipping; therefore, this genotype would influence CYP2C93-mediated drug metabolism. SV1 was expressed in 6 of 11 rhesus monkeys and 1 of 8 cynomolgus monkeys, but the SV1 in the cynomolgus monkey was nonfunctional due to a rare null genotype (c.102T>del). These results suggest that CYP2C93 can play roles as a drug-metabolizing enzyme in rhesus monkeys (not in cynomolgus monkeys), although its relative contribution to drug metabolism has yet to be validated.

    DOI: 10.1371/journal.pone.0016923

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Murayama Norie, Yamazaki Hiroshi .  CYP1D1, pseudogenized in human, is expressed and encodes a functional drug-metabolizing enzyme in cynomolgus monkey. .  Biochem Pharmacol81 ( 3 ) 442 - 450   2011年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cytochrome P450 (P450 or CYP) 1 family consists of the CYP1A, CYP1B, CYP1C, and CYP1D subfamilies. In the human genome, CYP1A1, CYP1A2, and CYP1B1 are expressed and encode functional enzymes, whereas CYP1D1P (formerly known as CYP1A8P) is present as a pseudogene due to five nonsense mutations in the putative coding region. In this study, we identified CYP1D1 cDNA, highly identical (nearly 95%) to human CYP1D1P sequence, in cynomolgus monkey, a species frequently used in drug metabolism studies due to its evolutionary closeness to human. The amino acid sequence deduced from cynomolgus monkey CYP1D1 cDNA shared the high sequence identity (91%) with human CYP1D1P (postulated from the gene sequence), and the highest sequence identity (44-45%) with CYP1A1 and CYP1A2 among cynomolgus monkey P450s. CYP1D1 mRNA was most abundantly expressed in liver, followed by kidney, and jejunum. The hepatic expression level of CYP1D1 mRNA was comparable to that of CYP1A1 mRNA and much higher than that of CYP1A2 mRNA. CYP1D1 was barely detectable in immunoblots of cynomolgus monkey liver. Cynomolgus monkey CYP1D1 mRNA was induced in primary hepatocytes with omeprazole. Cynomolgus monkey CYP1D1 protein heterologously expressed in Escherichia coli catalyzed ethoxyresorufin O-deethylation and caffeine 8-hydroxylation, which CYP1As also catalyze. Finally, no nonsense mutations, corresponding to those found in human CYP1D1P, were found in the 20 cynomolgus monkeys and 10 rhesus monkeys used in this study. These results suggest that CYP1D1 plays a role as a functional, drug-metabolizing enzyme in cynomolgus monkey liver.

    DOI: 10.1016/j.bcp.2010.11.003

    PubMed

  • Ise Ryota, Kondo Satoshi, Kato Hiroto, Imai Noritaka, Akiyama Hideo, Iwasaki Kazuhide, Yamazaki Hiroshi, Uno Yasuhiro .  Expression of cytochromes p450 in fetal, infant, and juvenile liver of cynomolgus macaques. .  Drug Metab Pharmacokinet26 ( 6 ) 621 - 626   2011年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Preclinical data of fetal, infant, and juvenile animals are important for the prediction of drug toxicity in fetuses and children. However, expression of drug-metabolizing enzymes, including cytochromes P450 (CYPs), have not been fully investigated in fetal, infant, or juvenile liver of the cynomolgus macaque, an animal species important for preclinical studies. In this study, hepatic expression of 20 cynomolgus macaque CYPs (mfCYPs) in the CYP1-4 subfamilies that are relevant to drug metabolism was measured in fetuses, infants, and juveniles using DNA microarrays. Expression of most mfCYPs, including those moderately or abundantly expressed in postnatal livers such as mfCYP2A23, mfCYP2A24, mfCYP2B6, mfCYP2C9, mfCYP2C19, mfCYP2C76, mfCYP2D17, mfCYP2E1 mfCYP3A4, and mfCYP3A5, was much less abundant in fetal livers, but increased substantially after birth. In contrast, expression of mfCYP2C8 in fetal livers was not substantially different from postnatal livers. Since human CYP3A7 is expressed more abundantly in fetal livers than in adult livers, mfCYP3A7, an ortholog of human CYP3A7, was analyzed by quantitative polymerase chain reaction. Expression of mfCYP3A7 in fetal livers was much lower than that in postnatal livers, and greatly increased after birth, unlike the expression of human CYP3A7. These results indicate that expression of most mfCYPs examined was low in fetal livers, but increased greatly in postnatal livers, with a few exceptions such as mfCYP2C8.

    DOI: 10.2133/dmpk.DMPK-11-NT-057

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Yamazaki Hiroshi .  Discovery of genetic variants in CYP1D1: implication for functional integrity of CYP1D1 in cynomolgus macaques and rhesus macaques. .  Drug Metab Pharmacokinet26 ( 6 ) 627 - 631   2011年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The cytochrome P450 (CYP) 1 family consists of the CYP1A, CYP1B, CYP1C, and CYP1D subfamilies. In humans, CYP1A1, CYP1A2, and CYP1B1 are expressed and encode functional enzymes, whereas CYP1D1P (formerly known as CYP1A8P) is present as a pseudogene as a result of five nonsense mutations in exon 2 and exon 7 of the putative coding region. We previously identified CYP1D1 in macaques and found that it was expressed and functional in liver. Moreover, the nonsense mutations in exon 2 and exon 7 were not found in the 20 cynomolgus macaques and 10 rhesus macaques analyzed in that previous study. These results raised the possibility that CYP1D1 is a functional gene in macaques; however, the possibility that nonsense mutations are present in other exons cannot be excluded. In this study, we sought to identify genetic variants of CYP1D1 in 63 cynomolgus macaques and 30 rhesus macaques; we did not find nonsense mutations in any coding exon of the animals analyzed. Moreover, 15 of the 63 cynomolgus macaques were analyzed by quantitative polymerase chain reaction, confirming hepatic expression of CYP1D1 in all 15 animals. These results suggest that CYP1D1 is most likely functional in cynomolgus macaques and rhesus macaques.

    DOI: 10.2133/dmpk.DMPK-11-NT-026

    PubMed

  • Ohtsuka Tatsuyuki, Yoshikawa Takahiro, Kozakai Kazumasa, Tsuneto Yumi, Uno Yasuhiro, Utoh Masahiro, Yamazaki Hiroshi, Kume Toshiyuki .  Alprazolam as an in vivo probe for studying induction of CYP3A in cynomolgus monkeys., Species difference between cynomolgus monkeys and humans on cytochromes P450 2D and 3A-dependent drug oxidation activities in liver microsomes .  Drug Metab Dispos38, 57 ( 10, 2 ) 1806, 164 - 1813, 170   2010年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Induction of the cytochrome P450 (P450) enzyme is a major concern in the drug discovery processes. To predict the clinical significance of enzyme induction, it is helpful to investigate pharmacokinetic alterations of a coadministered drug in a suitable animal model. In this study, we focus on the induction of CYP3A, which is involved in the metabolism of approximately 50% of marketed drugs and is inducible in both the liver and intestine. As a marker substrate for CYP3A activity, alprazolam (APZ) was selected and characterized using recombinant CYP3A enzymes expressed in Escherichia coli. Both human CYP3A4 and its cynomolgus P450 ortholog predominantly catalyzed APZ 4-hydroxylation with sigmoidal kinetics. When administered intravenously and orally to cynomolgus monkeys, APZ had moderate clearance; its first-pass extraction ratio after oral dosing was estimated to be 0.09 in the liver and 0.45 in the intestine. Pretreatment with multiple doses of rifampicin (20 mg/kg p.o. for 5 days), a known CYP3A inducer, significantly decreased plasma concentrations of APZ after intravenous and oral administrations (0.5 mg/kg), and first-pass extraction ratios were increased to 0.39 in the liver and 0.63 in the intestine. The results were comparable to those obtained in clinical drug-drug interaction (DDI) reports related to CYP3A induction, although the rate of recovery of CYP3A activity seemed to be slower than rates estimated in clinical studies. In conclusion, pharmacokinetic studies using APZ as a probe in monkeys may provide useful information regarding the prediction of clinical DDIs due to CYP3A induction.

    DOI: 10.1124/dmd.110.032656

    PubMed

  • Uno Yasuhiro, Uehara Shotaro, Kohara Sakae, Murayama Norie, Yamazaki Hiroshi .  Cynomolgus monkey CYP2D44 newly identified in liver, metabolizes bufuralol, and dextromethorphan. .  Drug Metab Dispos38 ( 9 ) 1486 - 1492   2010年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The cynomolgus monkey is used in drug metabolism studies, because of its evolutionary closeness to human, including cytochrome P450. Cynomolgus monkey CYP2D17, highly homologous to human CYP2D6, has been identified and characterized. Here, we report characterization of another CYP2D, CYP2D44, identified in cynomolgus monkey liver. The CYP2D44 cDNA contained an open reading frame of 497 amino acids sharing high sequence identity (87-93%) with other primate CYP2Ds. CYP2D44 mRNA was predominantly expressed in liver, similar to CYP2D17 mRNA. CYP2D17 and CYP2D44 form a gene cluster in the genome, similar to human CYP2Ds. Metabolic assays of the CYP2D17 and CYP2D44 proteins heterologously expressed in Escherichia coli indicated that CYP2D44 metabolized human CYP2D6 substrates, bufuralol and dextromethorphan (bufuralol 1'-hydroxylation and dextromethorphan O-demethylation) but to a lesser extent than CYP2D17. Kinetic analysis of dextromethorphan metabolism indicated that the apparent K(m) and V(max) of CYP2D17 and CYP2D44 catalyzed O-demethylation were similar, and, the V(max) values of CYP2D17 and CYP2D44 catalyzed N-demethylation (which human CYP2D6 catalyzes much less effectively) were similar, but the apparent K(m) of the CYP2D44 reaction was higher. Western blot analysis showed that CYP2D proteins were expressed in cynomolgus and rhesus monkey liver as well as in human and marmoset liver. Similar to CYP2D6, CYP2D44 copy number varied among the eight cynomolgus monkeys and four rhesus monkeys used in this study. These results indicated that CYP2D44, together with CYP2D17, had functional characteristics similar to those of human CYP2D6 but measurably differed in dextromethorphan N-demethylation, suggesting its importance for CYP2D-dependent drug metabolism in macaque.

    DOI: 10.1124/dmd.110.033274

    PubMed

  • Uehara Shotaro, Murayama Norie, Yamazaki Hiroshi, Uno Yasuhiro .  A novel CYP2A26 identified in cynomolgus monkey liver metabolizes coumarin. .  Xenobiotica40 ( 9 ) 621 - 629   2010年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A novel cytochrome P450 (CYP), CYP2A26, was identified and characterized in cynomolgus monkey, one of the animal species used in preclinical studies. Deduced amino acid sequences of CYP2A26 cDNA showed high sequence identities (91-95%) with cynomolgus monkey CYP2A23 and CYP2A24, and human CYP2A6 and CYP2A13. Phylogenetic analysis showed that macaque CYP2As (CYP2A26, CYP2A23, and CYP2A24) were most closely clustered with human CYP2As, unlike CYP2As of dog, rat, and mouse (other species also used in drug metabolism). Quantitative polymerase chain reaction analysis showed that CYP2A26 mRNA, along with CYP2A23 and CYP2A24 mRNAs, was expressed predominantly in the liver, where CYP2A proteins were also detected by immunoblotting. Drug-metabolizing assays using the CYP2A26 protein heterologously expressed in Escherichia coli indicated that CYP2A26 catalyzed coumarin 7-hydroxylation with its apparent K(m) lower than that of CYP2A24, but similar to those of CYP2A6 and CYP2A23. These results suggest an evolutionary closeness and functional similarity of cynomolgus monkey CYP2A26 (together with CYP2A23 and CYP2A24) to human CYP2A6, and its functional role as a drug-metabolizing enzyme in the liver.

    DOI: 10.3109/00498254.2010.501118

    PubMed

  • Osada Naoki, Uno Yasuhiro, Mineta Katsuhiko, Kameoka Yosuke, Takahashi Ichiro, Terao Keiji .  Ancient genome-wide admixture extends beyond the current hybrid zone between Macaca fascicularis and M. mulatta. .  Mol Ecol19 ( 14 ) 2884 - 2895   2010年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Macaca fascicularis and Macaca mulatta are two of the most commonly used laboratory macaques, yet their genetic differences at a genome-wide level remain unclear. We analysed the multilocus DNA sequence data of 54 autosomal loci obtained from M. fascicularis samples from three different geographic origins and M. mulatta samples of Burmese origin. M. fascicularis shows high nucleotide diversity, four to five times higher than humans, and a strong geographic population structure between Indonesian-Malaysian and Philippine macaques. The pattern of divergence and polymorphism between M. fascicularis and M. mulatta shows a footprint of genetic exchange not only within their current hybrid zone but also across a wider range for more than 1 million years. However, genetic admixture may not be a random event in the genome. Whereas randomly selected genic and intergenic regions have the same evolutionary dynamics between the species, some cytochrome oxidase P450 (CYP) genes (major chemical metabolizing genes and potential target genes for local adaptation) have a significantly larger species divergence than other genes. By surveying CYP3A5 gene sequences of more than a hundred macaques, we identified three nonsynonymous single nucleotide polymorphisms that were highly differentiated between the macaques. The mosaic pattern of species divergence in the genomes may be a consequence of genetic differentiation under ecological adaptation and may be a salient feature in the genomes of nascent species under parapatry.

    DOI: 10.1111/j.1365-294X.2010.04687.x

    PubMed

  • Chowdhury Goutam, Murayama Norie, Okada Yusuke, Uno Yasuhiro, Shimizu Makiko, Shibata Norio, Guengerich F Peter, Yamazaki Hiroshi .  Human liver microsomal cytochrome P450 3A enzymes involved in thalidomide 5-hydroxylation and formation of a glutathione conjugate. .  Chem Res Toxicol23 ( 6 ) 1018 - 1024   2010年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    (R)-Thalidomide was oxidized to 5-hydroxythalidomide and 5'-hydroxythalidomide by NADPH-fortified liver microsomes from humans and monkeys. (R)-Thalidomide was hydroxylated more efficiently than (S)-thalidomide. Recombinant human P450s 3A4, 3A5, and 3A7 and monkey P450s 3A8 and 3A5 (coexpressed with NADPH-P450 reductase in bacterial membranes) also catalyzed (R)-thalidomide 5-hydroxylation. Purified human P450s 2C19, 3A4, and 3A5 mediated (R)-thalidomide 5-hydroxylation at similar rates in reconstituted systems. P450 2C19 showed a rather nonsaturable substrate-velocity curve; however, P450s 3A4 and 3A5 showed sigmoidal curves. P450 also oxidized 5-hydroxythalidomide to an epoxide or dihydroxy compound. Liquid chromatography-mass spectrometry analysis revealed the formation of a glutathione conjugate from (R)- and (S)-5-hydroxythalidomide, catalyzed by liver microsomal P450s 3A4 and 3A5 in the presence of glutathione (assigned as a conjugate of 5-hydroxythalidomide formed on the phenyl ring). These results indicate that human P450s 3A4 and 3A5 mediate thalidomide 5-hydroxylation and further oxidation leading to a glutathione conjugate, which may be of relevance in the pharmacological and toxicological actions of thalidomide.

    DOI: 10.1021/tx900367p

    PubMed

  • Uno Yasuhiro, Matsushita Akinori, Osada Naoki, Uehara Shotaro, Kohara Sakae, Nagata Ryoichi, Fukuzaki Koichiro, Utoh Masahiro, Murayama Norie, Yamazaki Hiroshi .  Genetic variants of CYP3A4 and CYP3A5 in cynomolgus and rhesus macaques. .  Drug Metab Dispos38 ( 2 ) 209 - 214   2010年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus and rhesus macaques are frequently used in preclinical trials due to their close evolutionary relationships to humans. We conducted an initial screening for genetic variants in cynomolgus and rhesus macaque genes orthologous to human CYP3A4 and CYP3A5. Genetic screening of 78 Indochinese and Indonesian cynomolgus macaques and 34 Chinese rhesus macaques revealed a combined total of 42 CYP3A4 genetic variants, including 12 nonsynonymous variants, and 34 CYP3A5 genetic variants, including nine nonsynonymous variants. Four of these nonsynonymous variants were located at substrate recognition sites or the heme-binding region, domains essential for protein function, including c.886G>A (V296M) and c.1310G>A (S437N) in CYP3A4 and c.1437C>G (N479K) and c.1310G>C (T437S) in CYP3A5. The mutant proteins of these genetic variants were expressed in Escherichia coli and purified. Metabolic activity of these proteins measured using midazolam and nifedipine as substrates showed that none of these protein variants substantially influences the drug-metabolizing capacity of CYP3A4 or CYP3A5 protein. In Indonesian cynomolgus macaques, we also found IVS3+1delG in CYP3A4 and c.625A>T in CYP3A5, with which an intact protein cannot be produced due to a frameshift generated. Screening additional genomes revealed that two of 239 animals and three of 258 animals were heterozygous for IVS3+1delG of CYP3A4 and c.625A>T of CYP3A5, respectively. Some genetic variants were unevenly distributed between Indochinese and Indonesian cynomolgus macaques and between cynomolgus and rhesus macaques. Information on genetic diversity of macaque CYP3A4 and CYP3A5 presented here could be useful for successful drug metabolism studies conducted in macaques.

    DOI: 10.1124/dmd.109.029710

    PubMed

  • Uno Yasuhiro, Fujino Hideki, Iwasaki Kazuhide, Utoh Masahiro .  Macaque CYP2C76 encodes cytochrome P450 enzyme not orthologous to any human isozymes. .  Curr Drug Metab11 ( 2 ) 142 - 152   2010年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkey is used in the study of drug metabolism and toxicity due to its evolutionary closeness to human as compared with other non- human primate species. However, it has become certain that drug metabolism in monkeys is different than in humans. Such species differences have not been fully investigated at a molecular level largely due to the scarcity of information on drug-metabolizing enzyme genes. In cynomolgus monkey, we have identified cDNAs for 21 kinds of cytochromes P450 (CYPs), among which CYP2C76 does not correspond to any human CYP isozymes and is partly responsible for the difference in pitavastatin metabolism between cynomolgus monkey and human. In cynomolgus monkey CYP2C76, we identified numerous genetic variants including a null genotype. Heterozygotes for this null genotype are expected to be poor metabolizers in CYP2C76-mediated drug metabolism. To provide new clues to CYP2C76 function, here, we have taken advantage of sequence information that has been recently deposited to public databases to assess the presence of CYP2C76 orthologs in primate species. In this assessment, we found the CYP2C76 cDNA sequence in rhesus monkey, and a gene sequence highly homologous to cynomolgus monkey CYP2C76 in the marmoset and orangutan genomes, raising the possibility that CYP2C76 could also play a role in these primate species. This review paper gives an overview of CYP2C76 from isolation to molecular characterization, and its implication in drug metabolism.

    PubMed

  • Uno Yasuhiro, Matsuno Kiyomi, Nakamura Chika, Utoh Masahiro, Yamazaki Hiroshi .  Identification and characterization of CYP2C18 in the cynomolgus macaque (Macaca fascicularis). .  J Vet Med Sci72 ( 2 ) 225 - 228   2010年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The macaque is widely used for investigation of drug metabolism due to its evolutionary closeness to the human. However, the genetic backgrounds of drug-metabolizing enzymes have not been fully investigated; therefore, identification and characterization of drug-metabolizing enzyme genes are important for understanding drug metabolism in this species. In this study, we isolated and characterized a novel cytochrome P450 2C18 (CYP2C18) cDNA in cynomolgus macaques. This cDNA was highly homologous (96%) to human CYP2C18 cDNA. Cynomolgus CYP2C18 was preferentially expressed in the liver and kidney. Moreover, a metabolic assay using cynomolgus CYP2C18 protein heterologously expressed in Escherichia coli revealed its activity toward S-mephenytoin 4'-hydroxylation. These results suggest that cynomolgus CYP2C18 could function as a drug- metabolizing enzyme in the liver.

    DOI: 10.1292/jvms.09-0341

    PubMed

  • Iwasaki Kazuhide, Murayama Norie, Koizumi Ryo, Uno Yasuhiro, Yamazaki Hiroshi .  Comparison of cytochrome P450 3A enzymes in cynomolgus monkeys and humans. .  Drug Metab Pharmacokinet25 ( 4 ) 388 - 391   2010年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Drug metabolizing activities of cytochromes P450 (P450s, or CYPs) 3A4 and 3A5 in liver microsomes from the cynomolgus monkey [Macaca fascicularis (mf)] were investigated and compared with those of human P450 3A enzymes. Low activities for dealkylation of ethoxyresorufin and pentoxyresorufin were seen in recombinant monkey mfCYP3A4 and mfCYP3A5 and in recombinant human CYP3A4 and CYP3A5 expressed in bacterial membranes. Hydroxylation activities of mfCYP3A4 and mfCYP3A5 toward coumarin, paclitaxel, diclofenac, flurbiprofen, and S-mephenytoin were below detectable levels, as was also true for CYP3A4 and CYP3A5. Monkey mfCYP3A5 and mfCYP3A4 were highly active in bufuralol 1'-hydroxylation. mfCYP3A5 was efficient at dextromethorphan O-demethylation, although human CYP3A5 was unable to catalyze this reaction. Apparent bufuralol 1'-hydroxylation and dextromethorphan O-demethylation activities of monkey liver microsomes were higher than those of human liver microsomes, possibly because of contributions of mfCYP3A5 to these P450 2D-dependent drug oxidations. mfCYP3A5 and CYP3A5 catalyzed midazolam 1'-hydroxylation at a low substrate concentration more efficiently than the corresponding CYP3A4. mfCYP3A5 had higher testosterone 6beta-hydroxylase activity than mfCYP3A4, but the reverse relationship was observed in oxidation of nifedipine and hydroxylation of dexamethasone. These results demonstrate that monkey P450 3A enzymes have similar substrate selectivity to that of human P450 3A enzymes, but exhibit wider substrate selectivity toward P450 2D substrates.

    PubMed

  • Nakanishi Yasuharu, Matsushita Akinori, Matsuno Kiyomi, Iwasaki Kazuhide, Utoh Masahiro, Nakamura Chika, Uno Yasuhiro .  Regional distribution of cytochrome p450 mRNA expression in the liver and small intestine of cynomolgus monkeys. .  Drug Metab Pharmacokinet25 ( 3 ) 290 - 297   2010年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The cynomolgus monkey is used to study drug metabolism because of its evolutionary closeness to humans. Despite their importance, regional distribution of cytochrome P450 (CYP) enzymes including CYP3As in the liver and small intestine, the major sites of drug metabolism, has not been fully investigated in cynomolgus monkeys. In this study, we measured mRNA expression levels of 14 CYPs in the CYP1, 2, and 3 subfamilies, including orthologs of human CYP3A4 and CYP3A5, in the liver and small intestine of cynomolgus monkeys. Expression levels of each CYP mRNA in various regions of the liver were quantified and comparisons were made between the right lobe, quadrate lobe, left medial lobe, left lateral lobe, and caudate lobe and with four different sections of the right lobe. In the small intestine, the same mRNAs were measured in the duodenum and six different sections from the proximal jejunum to the distal ileum. Expression levels of the CYP mRNAs were not substantially different between liver samples, but varied between the different sections of the small intestine, including CYP3A4. These results suggest that analysis of distinct sections is required for a better understanding of cynomolgus monkey CYPs in the small intestine.

    PubMed

  • Uno Yasuhiro, Matsuno Kiyomi, Nakamura Chika, Utoh Masahiro, Yamazaki Hiroshi .  Identification and characterization of CYP2B6 cDNA in cynomolgus macaques (Macaca fascicularis). .  J Vet Med Sci71 ( 12 ) 1653 - 1656   2009年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cytochrome P450 2B6 (CYP2B6), an important drug-metabolizing enzyme, is involved in the metabolism of prescribed drugs in humans. Despite its importance, cDNA for a CYP2B6 ortholog has not been identified and characterized in cynomolgus macaques, which are frequently used in preclinical studies. In this study, cDNA highly homologous to human CYP2B6 was cloned from the cynomolgus macaque liver. This cDNA contained an open reading frame of 491 amino acids and functional domains characteristic for CYP protein, such as substrate recognition sites and a heme-binding region. Cynomolgus CYP2B6 was expressed predominantly in the liver with some extra-hepatic expression among 10 tissues. Moreover, cynomolgus CYP2B6 revealed activities toward testosterone 16beta- hydroxylation and bupropion hydroxylation. These results suggest that cynomolgus CYP2B6 has a functional role in the liver.

    DOI: 10.1292/jvms.001653

    PubMed

  • Uno Yasuhiro, Matsuno Kiyomi, Nakamura Chika, Utoh Masahiro, Yamazaki Hiroshi .  Cloning, expression, and characterization of CYP3A43 cDNA in cynomolgus macaque (Macaca fascicularis). .  Drug Metab Lett3 ( 4 ) 228 - 233   2009年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus macaques are frequently used in drug metabolism studies due to their evolutionary closeness to humans. Despite their importance, genes encoding drug-metabolizing enzymes have not been fully identified in this species. In this study, the cDNA orthologous to human cytochrome P450 3A43 (CYP3A43) was isolated. Deduced amino acids of this cDNA had a high sequence identity ( approximately 95%) to human CYP3A43 cDNA and contained characteristic motifs for CYP3A proteins, heme-binding region and substrate recognition sites. Among 10 tissues analyzed, cynomolgus CYP3A43 was expressed in liver, adrenal gland, and lung, with the highest expression seen in liver. Cynomolgus CYP3A43 protein heterologously expressed in Escherichia coli exhibited metabolic activity toward midazolam 1'-hydroxylation. These results indicated that cynomolgus CYP3A43 was expressed in liver and encoded a functional drug-metabolizing enzyme, and could contribute to overall drug metabolism in cynomolgus macaque liver if expressed as a protein.

    PubMed

  • Iwasaki K, Uno Y .  Cynomolgus monkey CYPs: a comparison with human CYPs. .  Xenobiotica39 ( 8 ) 578 - 581   2009年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Characteristics of twelve cytochromes P450 (CYPs) from cynomolgus monkeys were compared with those of human CYPs that play an important role in drug metabolism. Eleven members of CYP1A, CYP2A, CYP2C, CYP2D, CYP2E, and CYP3A subfamilies from cynomolgus monkeys exhibited a high degree of homologies (more than 90%) in cDNA and amino acid sequences with corresponding human CYPs, and catalysed typical reactions of corresponding human CYPs. One member of the cynomolgus monkey CYP2C subfamily, CYP2C76, exhibited a lower homology (around 70%) in amino acid sequences with other cynomolgus monkey and human CYP2C subfamilies. CYP2C76 catalysed typical CYP2C substrates with low activities, and has not been found in humans. CYPs identified in cynomolgus monkeys were similar to CYP1A1, CYP1A2, CYP2A6, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, and CYP3A5 in humans. These results indicate that cynomolgus monkeys express CYPs similar to human CYPs that are important in drug metabolism.

    DOI: 10.1080/00498250903003135

    PubMed

  • Uno Yasuhiro, Ohuchi Tsukasa, Uehara Shotaro, Kito Go, Kamataki Tetsuya, Nagata Ryoichi .  Sex-related differences in the expression of mfGSTA2, a novel GST identified in cynomolgus monkey (Macaca fascicularis). .  Drug Metab Dispos37 ( 3 ) 453 - 456   2009年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Glutathione S-transferase (GST) is one of the most important phase II drug-metabolizing enzymes, catalyzing the conjugation of electrophilic substrates to glutathione. Unlike in humans, a surprisingly limited number of GST genes have been identified in monkeys. The identification of additional GST genes in this model system would prove to be advantageous, because monkeys remain an important predictor of drug effects and toxicities in humans during preclinical studies. In this study, we report the identification and characterization of the following six cDNAs in cynomolgus monkeys: mfGSTA1, mfGSTA2, mfGSTM5, mfMGST1, mfGSTO1, and mfGSTZ1. These cDNAs encode GSTs highly homologous (approximately 95%) to human GST cDNAs. Among these, the mfGSTA1, mfGSTM5, mfMGST1, mfGSTO1, and mfGSTZ1 cDNAs correspond to a single human GST counterpart, whereas the mfGSTA2 cDNA is highly similar to human GSTA1 and GSTA2 cDNAs. An analysis of tissue samples indicates that these GST genes are predominantly expressed in the liver along with some extrahepatic expression as determined by real-time reverse transcriptase- polymerase chain reaction. It is interesting to note that mfGSTA2 is significantly differentially expressed between males and females in the jejunum, where a striking 8-fold higher expression level is observed in males. These results suggest that a potential sex difference in the metabolism of drugs may be mediated by mfGSTA2. This also provides a basis for the investigation of sex-dependent drug metabolism in monkeys.

    DOI: 10.1124/dmd.108.023747

    PubMed

  • Uno Yasuhiro, Sakuraba Hiroko, Uehara Shotaro, Kumano Takayuki, Matsuno Kiyomi, Nakamura Chika, Kito Go, Kamataki Tetsuya, Nagata Ryoichi .  A null allele impairs function of CYP2C76 gene in cynomolgus monkeys: a possible genetic tool for generation of a better animal model in drug metabolism. .  Drug Metab Dispos37 ( 1 ) 14 - 17   2009年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The monkey CYP2C76 gene does not correspond to any of the human CYP2C genes, and its enzyme is at least partly responsible for the species difference occasionally seen in drug metabolism between monkeys and humans. To establish a line and/or lines of monkeys that are expected to show metabolic patterns highly similar to humans, we set out to find monkeys that lacked CYP2C76 activity. By genetic screening of 73 monkeys and a database search of expressed sequence tags, we found a total of 10 nonsynonymous genetic variants in the coding region of CYP2C76, including a null genotype (c.449TG>A). Some of the variants were differently distributed between two animal groups originating from different geographical regions (Indochina and Indonesia). After screening 170 additional genomic samples, we identified a total of eight animals (six males and two females) that were heterozygous for c.449TG>A, which could be used for establishing a homozygous line. If the homozygotes show drug- metabolizing properties more similar to humans than wild-type monkeys, the homozygotes may serve as a better animal model for drug metabolism. The data presented in this article provide the essential genetic information to perform a successful study by using cynomolgus monkeys and present a possible tool to generate a better animal model for drug metabolism.

    DOI: 10.1124/dmd.108.023622

    PubMed

  • Nakajima Toshiaki, Ohtani Hitoshi, Satta Yoko, Uno Yasuhiro, Akari Hirofumi, Ishida Takafumi, Kimura Akinori .  Natural selection in the TLR-related genes in the course of primate evolution. .  Immunogenetics60 ( 12 ) 727 - 735   2008年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The innate immune system constitutes the front line of host defense against pathogens. Toll-like receptors (TLRs) recognize molecules derived from pathogens and play crucial roles in the innate immune system. Here, we provide evidence that the TLR-related genes have come under natural selection pressure in the course of primate evolution. We compared the nucleotide sequences of 16 TLR-related genes, including TLRs (TLR1-10), MYD88, TILAP, TICAM1, TICAM2, MD2, and CD14, among seven primate species. Analysis of the non-synonymous/synonymous substitution ratio revealed the presence of both strictly conserved and rapidly evolving regions in the TLR-related genes. The genomic segments encoding the intracellular Toll/interleukin 1 receptor domains, which exhibited lower rates of non- synonymous substitution, have undergone purifying selection. In contrast, TLR4, which carried a high proportion of non-synonymous substitutions in the part of extracellular domain spanning 200 amino acids, was found to have been the suggestive target of positive Darwinian selection in primate evolution. However, sequence analyses from 25 primate species, including eight hominoids, six Old World monkeys, eight New World monkeys, and three prosimians, showed no evidence that the pressure of positive Darwinian selection has shaped the pattern of sequence variations in TLR4 among New World monkeys and prosimians.

    DOI: 10.1007/s00251-008-0332-0

    PubMed

  • 宇野泰広,藤野秀樹,鬼頭剛,鵜藤雅裕,福崎好一郎,鎌滝哲也,永田良一 .  CYP2C76:薬物代謝において稀に見られるサルとヒトの種差の原因か .  日本予防医学会雑誌3 ( 1 ) 9 - 13   2008年4月査読

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    担当区分:筆頭著者   記述言語:日本語   掲載種別:研究論文(学術雑誌)  

  • Yasuhiro Uno .  Identification and characterization of CYP2C76 in cynomolgus monkeys .      2008年3月

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    記述言語:英語   掲載種別:学位論文(博士)  

  • Osada Naoki, Hashimoto Katsuyuki, Kameoka Yosuke, Hirata Makoto, Tanuma Reiko, Uno Yasuhiro, Inoue Itsuro, Hida Munetomo, Suzuki Yutaka, Sugano Sumio, Terao Keiji, Kusuda Jun, Takahashi Ichiro .  Large-scale analysis of Macaca fascicularis transcripts and inference of genetic divergence between M. fascicularis and M. mulatta., CYP2C76: is it responsible for species difference between monkeys and humans in drug metabolism? .  BMC Genomics9, 3 ( 1 ) 90, 9 - 90, 13   2008年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Cynomolgus macaques (Macaca fascicularis) are widely used as experimental animals in biomedical research and are closely related to other laboratory macaques, such as rhesus macaques (M. mulatta). We isolated 85,721 clones and determined 9407 full-insert sequences from cynomolgus monkey brain, testis, and liver. These sequences were annotated based on homology to human genes and stored in a database, QFbase http://genebank.nibio.go.jp/qfbase/. RESULTS: We found that 1024 transcripts did not represent any public human cDNA sequence and examined their expression using M. fascicularis oligonucleotide microarrays. Significant expression was detected for 544 (51%) of the unidentified transcripts. Moreover, we identified 226 genes containing exon alterations in the untranslated regions of the macaque transcripts, despite the highly conserved structure of the coding regions. Considering the polymorphism in the common ancestor of cynomolgus and rhesus macaques and the rate of PCR errors, the divergence time between the two species was estimated to be around 0.9 million years ago. CONCLUSION: Transcript data from Old World monkeys provide a means not only to determine the evolutionary difference between human and non-human primates but also to unveil hidden transcripts in the human genome. Increasing the genomic resources and information of macaque monkeys will greatly contribute to the development of evolutionary biology and biomedical sciences.

    DOI: 10.1186/1471-2164-9-90

    PubMed

  • Uno Yasuhiro, Suzuki Yutaka, Wakaguri Hiroyuki, Sakamoto Yoshiko, Sano Hitomi, Osada Naoki, Hashimoto Katsuyuki, Sugano Sumio, Inoue Ituro .  Expressed sequence tags from cynomolgus monkey (Macaca fascicularis) liver: a systematic identification of drug-metabolizing enzymes. .  FEBS Lett582 ( 2 ) 351 - 358   2008年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The liver, a major organ for drug metabolism, is physiologically similar between monkeys and humans. However, the paucity of identified genes has hampered a deep understanding of drug metabolism in monkeys. To provide such a genetic resource, 28655 expressed sequence tags (ESTs) were generated from a cynomolgus monkey liver full-length enriched cDNA library, which contained 23 unique ESTs homologous to human drug- metabolizing enzymes. Our comparative genomics approach identified nine lineage-specific candidate ESTs, including three drug-metabolizing enzymes, which could be important for understanding the physiological differences between monkeys and humans.

    DOI: 10.1016/j.febslet.2007.12.031

    PubMed

  • Uno Yasuhiro, Hosaka Shinya, Matsuno Kiyomi, Nakamura Chika, Kito Go, Kamataki Tetsuya, Nagata Ryoichi .  Characterization of cynomolgus monkey cytochrome P450 (CYP) cDNAs: is CYP2C76 the only monkey-specific CYP gene responsible for species differences in drug metabolism? .  Arch Biochem Biophys466 ( 1 ) 98 - 105   2007年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cynomolgus monkey CYP2C76 does not have a corresponding ortholog in humans, and it is at least partly responsible for differences in drug metabolism between monkeys and humans. To determine if CYP2C76 is the only monkey-specific CYP gene, we identified cynomolgus monkey cDNAs for CYP2A23, CYP2A24, CYP2E1, CYP2J2, CYP3A5, CYP3A8, CYP4A11, CYP4F3, CYP4F11, CYP4F12, and CYP4F45. These CYP cDNAs showed a high sequence identity (93-96%) to the homologous human CYP cDNAs. The monkey CYPs were preferentially expressed in liver among the analyzed tissues. Moreover, all five analyzed monkey CYPs (CYP2A23, CYP2A24, CYP2E1, CYP3A5, and CYP3A8) metabolized typical substrates for human CYPs in the corresponding subfamilies. These results suggest that these 11 monkey CYP cDNAs are closely related to the human CYP cDNAs and thus, unlike CYP2C76, are not apparent monkey-specific cDNAs.

    DOI: 10.1016/j.abb.2007.07.003

    PubMed

  • Uno Y, Kumano T, Kito G, Nagata R, Kamataki T, Fujino H .  CYP2C76-mediated species difference in drug metabolism: a comparison of pitavastatin metabolism between monkeys and humans. .  Xenobiotica37 ( 1 ) 30 - 43   2007年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The monkey is often used to predict metabolism of drugs in humans since it generally shows a metabolic pattern similar to humans. However, metabolic profiles different from humans are occasionally seen in monkeys for some drugs including pitavastatin. Recently, we have successfully identified a monkey-specific cytochrome P450 (CYP) 2C76, which possibly accounts for a species difference between monkeys and humans because of its sequence and functional uniqueness. The present study on the role of CYP2C76 and other monkey CYP2Cs in pitavastatin metabolism, as an example, has revealed that CYP2C76 is important for the metabolism of the lactone form, indicating a major role of CYP2C76 for the difference in the metabolism of pitavastatin and possibly other drugs between monkeys and humans. The current investigation on the involvement of CYP2C76 in the metabolism of other drugs is expected to reveal further the further importance of this monkey-specific drug-metabolizing enzyme.

    DOI: 10.1080/00498250600968275

    PubMed

  • Liu W-S, Wang A, Uno Y, Galitz D, Beattie C W, Ponce de Leon F A .  Genomic structure and transcript variants of the bovine DAZL gene. .  Cytogenet Genome Res116 ( 1-2 ) 65 - 71   2007年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The Deleted in AZoospermia Like (DAZL) gene is a member of the DAZ family and encodes an RNA-binding protein that is expressed in prenatal and postnatal germ cells of males and females. In the human, there are five highly-related members in the DAZ family, four (DAZ1-4) on the Y chromosome and one (DAZL) on an autosome (HSA3). Mutations in these genes have been linked to severe spermatogenic failure and infertility in men. In the present study, we have cloned and characterized the bovine DAZL (bDAZL) gene. The full-length bDAZL cDNA is predicted to encode a protein of 295 amino acids with an RNA recognition motif. The deduced protein sequence of bDAZL is 96 and 97% similar to human and mouse DAZL, respectively. Fluorescence in situ hybridization (FISH) maps bDAZL to the distal region on BTA1q. The bDAZL gene consists of 11 exons and 10 introns. A bDAZL pseudogene was identified on BTA16. Expression analysis of bDAZL in 13 different tissues by RT-PCR shows that two transcripts, variant 1 (2,996 bp) and variant 2 (1,373 bp), of the bDAZL gene are detected only in testis mRNA. The variants probably result from alternative RNA splicing as variant 1 contains an additional 1,623-bp insertion in the 3' UTR. Our results lay the groundwork for possible single nucleotide polymorphism (SNP) and functional studies of the DAZL gene in cattle.

    DOI: 10.1159/000097419

    PubMed

  • Uno Yasuhiro, Fujino Hideki, Kito Go, Kamataki Tetsuya, Nagata Ryoichi .  CYP2C76, a novel cytochrome P450 in cynomolgus monkey, is a major CYP2C in liver, metabolizing tolbutamide and testosterone. .  Mol Pharmacol70 ( 2 ) 477 - 486   2006年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Monkeys are widely used as a primate model to study drug metabolism because they generally show a metabolic pattern similar to humans. However, the paucity of information on cytochrome P450 (P450) genes has hampered a deep understanding of drug metabolism in the monkey. In this study, we report identification of the CYP2C76 cDNA newly identified in cynomolgus monkey and characterization of this CYP2C along with cynomolgus CYP2C20, CYP2C43, and CYP2C75. The CYP2C76 cDNA contains the open reading frame encoding a protein of 489 amino acids that are only approximately 80% identical to any human or monkey P450 cDNAs. Gene and protein expression of CYP2C76 was confirmed in the liver of cynomolgus and rhesus monkeys but not in humans or the great apes. Moreover, CYP2C76 is located at the end of the CYP2C gene cluster in the monkey genome, the region of which corresponds to the intergenic region adjacent to the CYP2C cluster in the human genome, strongly indicating that this gene does not have the ortholog in humans. Among the four CYP2C genes expressing predominantly in the liver, the expression level of CYP2C76 was the greatest, suggesting that CYP2C76 is a major CYP2C in the monkey liver. Assays for the capacity of CYP2C76 to metabolize drugs using several substrates typical for human CYP2Cs revealed that CYP2C76 showed unique metabolic activity. These results suggest that CYP2C76 contributes to overall drug-metabolizing activity in the monkey liver and might account for species difference occasionally seen in drug metabolism between monkeys and humans.

    DOI: 10.1124/mol.106.022673

    PubMed

  • Benkel Bernhard F, Nguyen Thuy, Uno Yasuhiro, Ponce de Leon F Abel, Hickey Donal A .  Structural organization and chromosomal location of the chicken alpha- amylase gene family. .  Gene362   117 - 124   2005年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Characterization of the Gallus gallus alpha-amylase gene family revealed that the chicken genome contains two distinct amy loci. One of the two loci is expressed in the chicken pancreas while cDNA clones for the second locus were detected in a library constructed from liver mRNA. Fluorescent in situ hybridization to chromosome spreads showed that the two loci are both located on chromosome 8 within the chicken genome. Moreover, each locus contains both an intact, expressed gene copy as well as a pseudogene. The expressed gene and the pseudogene are arranged in a divergent configuration in the pancreatic amy locus, while in the hepatic locus the intact gene and the pseudogene are arranged in tandem. The data suggest a complex pattern of evolution for the chicken amylase gene family which includes multiple gene duplication events, insertion/deletion events, as well as changes in spatial expression patterns.

    DOI: 10.1016/j.gene.2005.07.030

    PubMed

  • Uno Yasuhiro, Sakamoto Yoshiko, Yoshida Kenichi, Hasegawa Takashi, Hasegawa Yoshinori, Koshino Takeshi, Inoue Ituro .  Characterization of six base pair deletion in the putative HNF1-binding site of human PXR promoter. .  J Hum Genet48 ( 11 ) 594 - 597   2003年

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Pregnane X receptor (PXR) regulates transcription of drug metabolism genes such as CYP3A4 and MDR1. Several species of PXR transcripts have been reported, including hPAR-2 with an extended amino-terminus. Database search identified a 6-bp deletion at the putative HNF1 binding element on the proximal region flanking to the hPAR-2 transcription start site. Aspirin-induced asthma (AIA) is a typical drug-induced phenotype due to aspirin or nonsteroidal antiinflammatory drugs, and these drugs are metabolized by CYP2C9 and UGT1A6, which are regulated by PXR. We examined a possible association between the 6-bp deletion variant and AIA; 129 AIA patients and 117 controls were genotyped, and no allelic association was observed. Characterization of the hPAR-2 promoter revealed that the proximal region of 1.5-kb from the transcription start site conferred a promoter activity and that the 6-bp deletion diminished the activity. These results suggest that the putative HNF1 binding element is essential for the transcriptional activity of hPAR-2 and also, that substantial numbers of Japanese are in a deficient state. Because of the biological significance of the 6-bp deletion of PXR, the variant might potentially associate with as yet unknown phenotype.

    DOI: 10.1007/s10038-003-0076-5

    PubMed

▼全件表示

講演・口頭発表等

  • 宇野泰広,山崎浩史 .  カニクイザルにおける薬物代謝酵素の同定・解析 .  第48回日本毒性学会学術年会  2021年7月 

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    開催年月日: 2021年7月

    開催地:神戸  

  • 尾野由佳,岡松岳,杉山颯太,宇野泰広,生城真一,北澤多喜雄,寺岡宏樹 .  ネコにおける異物代謝に関わるシトクロームP450分子種の特徴 .  第163回日本獣医学会学術集会 

     詳細を見る

    開催年月日: 2020年9月

    記述言語:日本語   会議種別:ポスター発表  

    開催地:山口大学  

  • Shiori Honda, Tatsuki Fukami, Takuya Tsujiguchi, Yongjie Zhang, Masataka Nakano, Shotaro Uehara, Yasuhiro Uno, Hiroshi Yamazaki, and Miki Nakajima .  Species differences in hydrolase activities by CES and AADAC between cynomolgus monkeys, marmosets, and humans .  日本薬物動態学会第34年会  国際会議

     詳細を見る

    開催年月日: 2019年12月

    記述言語:英語   会議種別:ポスター発表  

    開催地:つくば  

  • Norie Murayama, Shotaro Uehara, Shu Kawamura, Miu Nishikawa, Shinichi Ikushiro, Yasuhiro Uno, and Hiroshi Yamazaki .  Functional and molecular characterization of UDP-glucuronosyltransferase 1A enzymes in common marmosets .  日本薬物動態学会第34年会 

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    開催年月日: 2019年12月

    記述言語:英語   会議種別:ポスター発表  

    開催地:つくば  

  • Yasuhiro Uno, Hiroshi Yamazaki .  Analysis of microRNAs as cytochrome P450 regulators in cynomolgus macaque .  日本薬物動態学会第34年会 

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    開催年月日: 2019年12月

    記述言語:英語   会議種別:ポスター発表  

    開催地:つくば  

  • 河村秀,村山典恵,小野関駿佑,石井裕,上原正太郎,宇野泰広,生城真一,山崎浩史 .  マーモセット肝UDP-グルクロン酸転移酵素 (UGT) 触媒機能のサルとヒトとの比較 .  日本薬学会第139年会 

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    開催年月日: 2019年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 田中佐季,村山典恵,加藤雅巳,上原正太郎,宇野泰広,山崎浩史 .  医薬品等の抱合反応を触媒するマーモセット肝グルタチオンS-転移酵素分子種 .  日本薬学会第139年会 

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    開催年月日: 2019年3月

    記述言語:日本語   会議種別:ポスター発表  

  • Yasuhiro Uno, Norie Murayama, Shinichi Ikushiro, and Hiroshi Yamazaki .  Characterization of UDP-glucuronosyltransferase 2B enzymes in cynomolgus macaques .  2018 International Meeting on 22nd MDO and 33rd JSSX(日本薬物動態学会第33年会/MDO国際合同学会)  国際会議

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    開催年月日: 2018年10月

    記述言語:英語   会議種別:ポスター発表  

  • Norie Murayama, Shotaro Uehara, Yu Ishii, Shunsuke Onozeki, Shu Kawamura, Shinichi Ikushiro, Yasuhiro Uno, and Hiroshi Yamazaki .  Activities of UDP-glucuronosyltransferase 1A/2B enzymes in marmoset liver microsomes: comparison with cynomolgus macaques and humans .  2018 International Meeting on 22nd MDO and 33rd JSSX(日本薬物動態学会第33年会/MDO国際合同学会)  国際会議

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    開催年月日: 2018年10月

    記述言語:英語   会議種別:ポスター発表  

  • Tsuyoshi Takahashi, Akira Oku, Yasuhiro Uno, Hiroshi Yamazaki, and Toshiyuki Kume .  Characterization of probe substrates for polymorphic organic-anion-transporting polypeptides (OATP/SLCO1B1) in cynomolgus monkeys .  2018 International Meeting on 22nd MDO and 33rd JSSX(日本薬物動態学会第33年会/MDO国際合同学会)  国際会議

     詳細を見る

    開催年月日: 2018年10月

    記述言語:英語   会議種別:ポスター発表  

  • Teruko Imai, Maori Tanaka, Yoshiyuki Igawa, Kayoko Ohura, Masakiyo Hosokawa, and Yasuhiro Uno .  A study of functional analyses for carboxylesterase type 2 isozymes in cynomolgus monkeys .  2018 International Meeting on 22nd MDO and 33rd JSSX(日本薬物動態学会第33年会/MDO国際合同学会)  国際会議

     詳細を見る

    開催年月日: 2018年10月

    記述言語:英語   会議種別:ポスター発表  

  • 宇野泰広,山崎浩史 .  カニクイザルにおけるチトクロムP450遺伝子の解析と薬物動態 .  第45回日本毒性学会学術年会  招待

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    開催年月日: 2018年7月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • Sachiko Sakai, Kenta Mizoi, Masato Takahashi, Yasuhiro Uno, Teruko Imai, and Masakiyo Hosokawa .  Species differences of mouse, cynomolgus monkey and human carboxylesterase using atorvastatin esters with various steric and electronic properties .  第32回日本薬物動態学会年会 

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    開催年月日: 2017年11月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno, Shinichi Ikushiro, Norie Murayama, and Hiroshi Yamazaki .  Identification and characterization of UDP-glucuronosyltransferase 2 transcripts in cynomolgus macaque .  第32回日本薬物動態学会年会 

     詳細を見る

    開催年月日: 2017年11月

    記述言語:英語   会議種別:ポスター発表  

  • Norie Murayama, Yu Ishii, Yasuhiro Uno, Shinichi Ikushiro, and Hiroshi Yamazaki .  UDP-glucuronosyltransferase (UGT) activities in cynomolgus monkey livers .  第32回日本薬物動態学会年会 

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    開催年月日: 2017年11月

    記述言語:英語   会議種別:ポスター発表  

  • Shinichi Ikushiro, Rika Takahira, Yuuka Masuyama, Miyu Nishikawa, Kaori Yasuda, Yasuhiro Uno, Norie Murayama, Hiroshi Yamazaki, and Toshiyuki Sakaki .  Construction of expression system of cynomolgus macaque UDP-glucuronosyltransferase 1A isoforms using budding yeast .  第32回日本薬物動態学会年会 

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    開催年月日: 2017年11月

    記述言語:英語   会議種別:ポスター発表  

  • Kayoko Ohura, Maori Tanaka, Kana Matsumoto, Yoshiyuki Igawa, Akiko Kasahara, Naoya Wada, Kazuishi Kubota, Yasuhiro Uno, and Teruko Imai .  Expression of carboxylesterase 2 isozymes in cynomolgus monkey intestine and their hydrolytic properties .  第32回日本薬物動態学会年会 

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    開催年月日: 2017年11月

    記述言語:英語   会議種別:ポスター発表  

  • 小原栄,和田郁人,泉良和,宇野泰広,橘敬祐,小林直之,和泉博之,斯波真理子 .  デジタルPCR を用いた人工核酸定量法の検討 .  日本核酸医薬学会第3回年会 

     詳細を見る

    開催年月日: 2017年7月

    記述言語:日本語   会議種別:ポスター発表  

  • 田中真織,井川佳之,大浦華代子,細川正清,宇野泰広,今井輝子 .  サルカルボキシルエステラーゼ2分子種の加水分解特性と臓器発現の個体差 .  日本薬剤学会第32年会 

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    開催年月日: 2017年5月

    記述言語:日本語   会議種別:ポスター発表  

  • 村山典恵,加藤雅巳,大越智子,宇野泰広,山崎浩史 .  カニクイザル肝グルタチオン S-転移酵素thetaクラス分子種の酵素機能解析 .  日本薬学会第137年会 

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    開催年月日: 2017年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 小原栄,泉良和,岡谷秀明,宇野泰広,鵜藤雅裕,和泉博之 .  ヒト由来細胞の動物生体内分布の評価-定量PCR法の比較- .  第16回日本再生医療学会総会 

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    開催年月日: 2017年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 石井さくら,上原正太郎,宇野泰広,井上貴史,佐々木えりか,山崎浩史 .  マーモセット,カニクイザルおよびヒト肝によるメトプロロール酸化的代謝の比較 .  日本薬学会第137年会 

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    開催年月日: 2017年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 上原正太郎,中西一志,宇野泰広,井上貴史,佐々木えりか,山崎浩史 .  マーモセットP450 3A酵素の臓器分布および薬物酸化酵素活性:ヒトP450 3Aとの類似性 .  日本薬学会第137年会 

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    開催年月日: 2017年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 結城友香子,上原正太郎,宇野泰広,井上貴史,佐々木えりか,山崎浩史 .  マーモセット,カニクイザルおよびヒト肝および小腸P450酵素によるテルフェナジンの酸化的代謝 .  日本薬学会第137年会 

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    開催年月日: 2017年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 上原正太郎,大塩徹,宇野泰広,井上貴史,佐々木えりか,山崎浩史 .  新規マーモセットP450 2F酵素の同定および機能解析 .  日本薬学会第137年会 

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    開催年月日: 2017年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 三浦智徳,草間崇史,三井満里奈,河野未来,平野貴弘,宇野泰広,鵜藤雅裕,清水万紀子,山崎浩史 .  チトクロム P450 2C9 遺伝子多型を有するカニクイザル個体のエファビレンツ血中動態推移予測 .  日本薬学会第137年会 

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    開催年月日: 2017年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 草間崇史,三浦智徳,三井満里奈,河野未来,平野貴弘,宇野泰広,鵜藤雅裕,清水万紀子,山崎浩史 .  チトクロムP450 2C19多型性を示すカニクイザル個体別のR-ワルファリン血中濃度推移予測 .  日本薬学会第137年会 

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    開催年月日: 2017年3月

    記述言語:日本語   会議種別:ポスター発表  

  • Kazuhide Iwasaki, Yusuke Kitsugi, Kanami Ikeda, Takahiro Yoshikawa, Shotaro Uehara, Yasuhiro Uno, Masahiro Utoh, and Hiroshi Yamazaki .  Effect of CYP2C9 genotype for pharmacokinetics of efavirenz in vivo in cynomolgus monkeys .  11th International ISSX Meeting  国際会議

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    開催年月日: 2016年12月

    記述言語:英語   会議種別:ポスター発表  

  • Shotaro Uehara, Yasuhiro Uno, Takashi Inoue, Erika Sasaki, and Hiroshi Yamazaki .  Identification and characterization of cytochrome P450 4F enzymes in marmosets .  11th International ISSX Meeting  国際会議

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    開催年月日: 2016年12月

    記述言語:英語   会議種別:ポスター発表  

  • 小原栄,和田郁人,宇野泰広,鵜藤雅裕,橘敬祐,小比賀聡,小林直之,和泉博之,斯波真理子 .  LCR(ligase chain reaction)法を用いた人工核酸定量法の開発 .  日本核酸医薬学会第2回年会 

     詳細を見る

    開催年月日: 2016年11月

    記述言語:日本語   会議種別:ポスター発表  

  • Norie Murayama, Kanako Yajima,Mikiko Hikawa, Yuu Ishii, Masaki Takada, Masami Kato, Kanami Shimura, Tomonori Miura, Yasuhiro Uno, Kazuhide Iwasaki, Masahiro Uto, and Hiroshi Yamazaki .  Roles of human cytochrome P450 forms in drug oxidations determined using pooled liver microsomes in which one form had been metabolically inactivated .  第31回日本薬物動態学会年会 

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    開催年月日: 2016年10月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno, Norie Murayama, Masahiro Utoh, and Hiroshi Yamazaki .  Systematic identification and characterization of arylamine N-acetyltransferases in cynomolgus macaque .  第31回日本薬物動態学会年会 

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    開催年月日: 2016年10月

    記述言語:英語   会議種別:ポスター発表  

  • Masahiro Utoh, Takashi Kusama, Tomonori Miura, Marina Mitsui, Mirai Kawano, Takahiro Hirano, Makiko Shimizu, Yasuhiro Uno, and Hiroshi Yamazaki .  Simulation for R-warfarin clearance associated with polymorphic cytochrome P450 2C19 in individual cynomolgus monkeys .  第31回日本薬物動態学会年会 

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    開催年月日: 2016年10月

    記述言語:英語   会議種別:ポスター発表  

  • Kanako Yajima, Norie Murayama, Yasuhiro Uno, Masahiro Utoh, Kazuhide Iwasaki, Hiroshi Yamazaki .  Determination of in vitro fraction metabolized (fm) values for human cytochrome enzymes P450 using Silensomes .  第31回日本薬物動態学会年会 

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    開催年月日: 2016年10月

    記述言語:英語   会議種別:ポスター発表  

  • 坂井知津香,岩野俊介,東恵理子,内田将史,小野寺純,宇野泰広,林亮司,杉山篤,中村和市,宮本庸平,山崎浩史 .  マイクロミニピッグにおける薬物動態関連機能分子の解析 .  第43回日本毒性学会学術年会 

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    開催年月日: 2016年6月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 今井輝子,井川佳之,西澤遥,大浦華代子,宇野泰広,細川正清 .  新規カニクイザル カルボキシルエステラーゼ2分子種の加水分解特性と肝臓および小腸における発現の個体差 .  日本薬学会第136年会 

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    開催年月日: 2016年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 小原栄,岡谷秀明,宇野泰広,鵜藤雅裕,和泉博之 .  ヒト由来細胞の動物生体内分布の評価法構築 .  第15回日本再生医療学会総会 

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    開催年月日: 2016年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 河野未来,上原正太郎,宇野泰広,井上貴史,鵜藤雅裕,佐々木えりか,山崎浩史 .  マーモセット肝のワルファリン酸化的代謝の立体選択性 .  日本薬学会第136年会 

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    開催年月日: 2016年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 清水万紀子,三井満里奈,河野未来,上原正太郎,宇野泰広,鵜藤雅裕,井上貴史,佐々木えりか,末水洋志,山崎浩史 .  ヒト肝細胞移植マウスおよびマーモセット生理学的薬物動態モデルを活用したチトクロムP450カクテル基質のヒト血中濃度推移予測 .  日本薬学会第136年会 

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    開催年月日: 2016年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 上原正太郎,岡本絵里子,宇野泰広,井上貴史,佐々木えりか,山崎浩史 .  新規マーモセットP450 2J酵素の同定および機能解析 .  日本薬学会第136年会 

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    開催年月日: 2016年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 上原正太郎,結城友香子,宇野泰広,井上貴史,佐々木えりか,山崎浩史 .  マーモセットおよびカニクイザル小腸P450酵素によるエバスチン酸化的代謝 .  日本薬学会第136年会 

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    開催年月日: 2016年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 鈴木貴子,上原正太郎,村山典恵,宇野泰広,矢島加奈子,鵜藤雅裕,井上貴史,佐々木えりか,山崎浩史 .  マーモセットあるいはカニクイザル肝細胞を用いた薬物暴露によるチトクロムP450誘導の評価 .  日本薬学会第136年会 

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    開催年月日: 2016年3月

    記述言語:日本語   会議種別:ポスター発表  

  • Yoshiyuki Igawa, You Nishizawa, Kayoko Ohura, Yasuhiro Uno, Hosokawa Masakiyo, and Teruko Imai .  Hydrolytic properties of two carboxylesterase 2 isozymes and their expression in individual cynomolgus monkey intestine .  第30回日本薬物動態学会年会 

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    開催年月日: 2015年11月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno and Hiroshi Yamazaki .  Analysis of cytochrome P450 regulators in cynomolgus macaque .  第30回日本薬物動態学会年会 

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    開催年月日: 2015年11月

    記述言語:英語   会議種別:ポスター発表  

  • Tsuyoshi Takahashi, Tatsuyuki Ohtsuka, Yasuhiro Uno, Hiroshi Yamazaki, and Toshiyuki Kume .  Time-dependent inhibition of OATP-mediated transport by cyclosporine A is reproduced in cynomolgus monkeys .  第30回日本薬物動態学会年会 

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    開催年月日: 2015年11月

    記述言語:英語   会議種別:ポスター発表  

  • Yusuke Kitsugi, Kanami Ikeda, Atsushi Matsuoka, Takaaki Tokunaga, Natsumi Tomida, Haruka Shirosawa, Takahiro Yoshikawa, Yasuhiro Uno, Kazuhide Iwasaki, Hiroshi Yamazaki, and Masahiro Utoh .  Pharmacokinetics of efavirenz and its metabolites after oral and intravenous administration of efavirenz to cynomolgus monkeys .  第30回日本薬物動態学会年会 

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    開催年月日: 2015年11月

    記述言語:英語   会議種別:ポスター発表  

  • 山崎浩史,宇野泰広 .  実験動物およびヒトの薬物酸化酵素活性の比較 .  第42回日本毒性学会学術年会  招待

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    開催年月日: 2015年6月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 高平梨可,宇野泰広,池中良徳,石塚真由美,西川美宇,榊利之,生城真一 .  出芽酵母発現系を用いたカニクイザル由来UDP-グルクロン酸転移酵素の異物抱合能の解析 .  第42回日本毒性学会学術年会 

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    開催年月日: 2015年6月

    記述言語:日本語   会議種別:ポスター発表  

  • Shinya Hosaka, Norie Murayama, Masahiro Satsukawa, Shotaro Uehara, Makiko Shimizu, Kazuhide Iwasaki, Shunsuke Iwano, Yasuhiro Uno, and Hiroshi Yamazaki .  Comprehensive evaluation for substrate selectivity of cynomolgus monkey cytochrome P450 2C enzymes .  19th International Conference on Cytochrome P450  国際会議

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    開催年月日: 2015年6月

    記述言語:英語   会議種別:ポスター発表  

  • Shotaro Uehara, Yasuhiro Uno, Norie Murayama, Makiko Shimizu, and Hiroshi Yamazaki .  Molecular cloning, expression and functional characterization of marmoset cytochrome P450 2A6 .  19th International Conference on Cytochrome P450  国際会議

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    開催年月日: 2015年6月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno, Shotaro Uehara, and Hiroshi Yamazaki .  Identification and analysis of CYP2B6 variants in cynomolgus and rhesus macaques .  19th International Conference on Cytochrome P450  国際会議

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    開催年月日: 2015年6月

    記述言語:英語   会議種別:ポスター発表  

  • Masahiro Utoh, Takahiro Yoshikawa, Makiko Shimizu, Kazuhide Iwasaki, Yasuhiro Uno, and Hiroshi Yamazaki .  In vivo R/S-warfarin metabolism mediated by polymorphic P450 2C19 in cynomolgus monkeys .  19th International Conference on Cytochrome P450  国際会議

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    開催年月日: 2015年6月

    記述言語:英語   会議種別:ポスター発表  

  • 宇野泰広,山崎浩史 .  カニクイザルにおけるチトクロームP450とグルクロン酸抱合酵素の解析 .  第42回日本毒性学会学術年会  招待

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    開催年月日: 2015年6月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 志田聡美,村山典恵,宇野泰広,山崎浩史 .  カニクイザル生理学的薬物動態モデルを活用したチトクロムP450 カクテル基質のヒト血中濃度推移予測 .  日本薬学会第135年会 

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    開催年月日: 2015年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 高平梨可,西川美宇,宇野泰広,池中良徳,石塚真由美,榊利之,生城真一 .  カニクイザル由来UDP-グルクロン酸転移酵素におけるポリフェノール類代謝能の解析 .  日本農芸化学会2015年度大会 

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    開催年月日: 2015年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 保坂信哉,村山典恵,薩川正広,清水万紀子,上原正太郎,岩﨑一秀,岩野俊介,宇野泰広,山崎浩史 .  カニクイザルシトクロムP450 2C9の基質特異性に関する検討 .  日本薬学会第135年会 

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    開催年月日: 2015年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 西山咲弥子,清水万紀子,上原正太郎,宇野泰広,山崎浩史 .  サリドマイド1次および2次酸化的代謝に関与するチトクロム P450 の種差 .  日本薬学会第135年会 

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    開催年月日: 2015年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 岡松岳,宇野泰広,打出毅,北澤多喜雄,寺岡宏樹 .  各種ネコ シトクロームP450発現タンパクの代謝活性 .  第37回日本分子生物学会年会 

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    開催年月日: 2014年11月

    記述言語:日本語   会議種別:ポスター発表  

  • 高平梨可,宇野泰広,池中良徳,石塚真由美,西川美宇,榊利之,生城真一 .  出芽酵母発現系を用いたカニクイザル由来UDP-グルクロン酸転移酵素の機能解析 .  第87回日本生化学会大会 

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    開催年月日: 2014年10月

    記述言語:日本語   会議種別:ポスター発表  

  • 西山咲弥子,清水万紀子,上原正太郎,宇野泰広,山崎浩史 .  サリドマイド代謝的活性化における霊長類チトクロム P450 の役割 .  第58回日本薬学会関東支部大会 

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    開催年月日: 2014年10月

    記述言語:日本語   会議種別:ポスター発表  

  • Hiroshi Yamazaki, Yasuhiro Uno, Mitsunori Shukuya, Takahiro Yoshikawa, Yasuka Matsumoto, Akinori Matsushita, Norie Murayama, Koichiro Fukuzaki, and Masahiro Utoh .  Warfarin metabolism mediated by polymorphic P450 2C19 in cynomolgus monkeys .  19th North American Regional ISSX Meeting and 29th JSSX Annual Meeting  国際会議

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    開催年月日: 2014年10月

    記述言語:英語   会議種別:ポスター発表  

  • 宇野泰広,村山典恵,山崎浩史 .  カニクイザル硫酸転移酵素の同定・解析 .  第157回日本獣医学会学術集会 

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    開催年月日: 2014年9月

    記述言語:日本語   会議種別:口頭発表(一般)  

  • 中西康晴, 中村稚加, 岩崎一秀, 鵜藤雅裕, 山崎浩史, 宇野泰広 .  カニクイザルの小腸におけるUDP-グルクロン酸転移酵素活性 .  第41回 日本毒性学会学術年会 

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    開催年月日: 2014年7月

    記述言語:日本語   会議種別:ポスター発表  

  • 岡松岳,川上圭,小松徹也,打出毅,宇野泰広,北澤多喜雄,寺岡宏樹 .  大腸菌発現系を用いたシトクローム P450 3Aのネコ、ヒト、イヌでの動物種差の検討 .  第28回日本中毒学会東日本地方会 

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    開催年月日: 2014年1月

    記述言語:日本語   会議種別:ポスター発表  

  • Kanako Yajima, Norie Murayama, Shotaro Uehara, Chika Nakamura, Yasuhiro Uno, Kazuhide Iwasaki, Hiroshi Yamazaki, and Masahiro Utoh .  Evaluation of cytochrome P450 1A, 2B, 2C, and 3A induction by drugs in cryopreserved human hepatocytes .  第28回日本薬物動態学会年会 

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    開催年月日: 2013年10月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuharu Nakanishi, Chika Nakamura, Kazuhide Iwasaki, Masahiro Utoh, Hiroshi Yamazaki, and Yasuhiro Uno .  Microsomal UDP-glucuronosyltransferase activities in cynomolgus monkey livers .  第28回日本薬物動態学会年会 

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    開催年月日: 2013年10月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno, Yoshinori Ikenaka, and Mayumi Ishizuka .  Systematic identification and characterization of UDP-glucuronosyltransferases in cynomolgus macaque .  第28回日本薬物動態学会年会 

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    開催年月日: 2013年10月

    記述言語:英語   会議種別:ポスター発表  

  • Takahiro Yoshikawa, Natsumi Oyama, Yusuke Kitsugi, Atsushi Matsuoka, Yoshiharu Hayashi, Takatoshi Nakamura, Yasuhiro Uno, Kazuhide Iwasaki, Takanori Sakai, Norie Murayama, Hiroshi Yamazaki, and Masahiro Utoh .  Pharmacokinetics of warfarin and its metabolites after oral administration of warfarin to cynomolgus monkeys (2) -In vivo metabolism of warfarin- .  第28回日本薬物動態学会年会 

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    開催年月日: 2013年10月

    記述言語:英語   会議種別:ポスター発表  

  • 宇野泰広,池中良徳,石塚真由美 .  カニクイザルUGT1Aの同定・解析 .  第156回日本獣医学会学術集会 

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    開催年月日: 2013年9月

    記述言語:日本語   会議種別:口頭発表(一般)  

  • 山崎美穂,清水万紀子,宇野泰広,小林由布子,村山典恵,岩崎一秀,鵜藤雅裕,山崎浩史 .  フラビン含有モノオキシゲナーゼが触媒する薬物酸化酵素活性のヒトと実験動物の種差 .  第133回日本薬学会年会 

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    開催年月日: 2013年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 村山典恵,宇野泰広,細井祥生,岩崎一秀,清水万紀子,鵜藤雅裕,山崎浩史 .  ヒト及びサル肝P450に認められるワルファリン代謝の立体選択性 .  第133回日本薬学会年会 

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    開催年月日: 2013年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 鵜藤雅裕,村山典恵,宇野泰広,小野瀬優維,清水万紀子,岩崎一秀,山崎浩史 .  カフェインからテオフィリンを生成するサル肝P450 .  第133回日本薬学会年会 

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    開催年月日: 2013年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 池田可奈実,村山典恵,松本和華,宿谷光則,余合美咲,杉山洋介,清水万紀子,宇野泰広,岩崎一秀,鵜藤雅裕,山崎浩史 .  ブタ、サルおよびヒト肝チトクロムP450 2Dおよび3A酵素活性 .  第133回日本薬学会年会 

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    開催年月日: 2013年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 山崎浩史,村山典恵,清水万紀子,宇野泰広 .  マーモセット肝チトクロムP450 2D酵素活性 .  第2回日本マーモセット研究会 

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    開催年月日: 2013年2月

    記述言語:日本語   会議種別:ポスター発表  

  • Kayoko Ohura, Seiya Fujihara, Yasuhiro Uno, Masakiyo Hosokawa, and Teruko Imai .  Enzymatic characteristics of monkey and human intestinal carboxylesterase 2 and its age-dependent expression .  第27回日本薬物動態学会年会 

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    開催年月日: 2012年11月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuharu Nakanishi, Hiroyuki Yamashita, Tsuyoshi Yoshikawa, Masahiro Utoh, Chika Nakamura, and Yasuhiro Uno .  Cytochromes P450 enzyme activities in the small intestine of cynomolgus monkeys from Cambodia, China, and Indonesia .  第27回日本薬物動態学会年会 

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    開催年月日: 2012年11月

    記述言語:英語   会議種別:ポスター発表  

  • Shotaro Uehara, Norie Murayama, Chika Nakamura, Hiroshi Yamazaki, and Yasuhiro Uno .  Expression of cytochrome P450 enzymes in small intestine microsomes of 35 cynomolgus monkeys .  第27回日本薬物動態学会年会 

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    開催年月日: 2012年11月

    記述言語:英語   会議種別:ポスター発表  

  • Kanako Yajima, Norie Murayama, Shotaro Uehara, Yoshikazu Izumi, Chika Nakamura, Yasuhiro Uno, Kazuhide Iwasaki, Hiroshi Yamazaki, and Masahiro Utoh .  Evaluation of cytochrome P450 induction by drugs in cryopreserved human hepatocytes and HepaRG cells .  第27回日本薬物動態学会年会 

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    開催年月日: 2012年11月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno, Norie Murayama, Mutsuki Kunori, and Hiroshi Yamazaki .  Systematic identification and characterization of glutathione S-transferases in cynomolgus macaques .  第27回日本薬物動態学会年会 

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    開催年月日: 2012年11月

    記述言語:英語   会議種別:ポスター発表  

  • Tadashi Ishida, Sachiyo Sudou, Yuma Hatano, Yasuhiro Uno,Teruko Imai, and Masakiyo Hosokawa .  Characterization and poperties of cynomolgus monkey carboxylesterase (CES) isozymes .  第27回日本薬物動態学会年会 

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    開催年月日: 2012年11月

    記述言語:英語   会議種別:ポスター発表  

  • Masahiro Utoh, Yasuhiro Uno, Tatsuyuki Ohtsuka, and Toshiyuki Kume .  Identification and analysis of drug transporters in cynomolgus macaques .  第27回日本薬物動態学会年会 

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    開催年月日: 2012年11月

    記述言語:英語   会議種別:ポスター発表  

  • 上原正太郎,村山典恵,中村稚加,山崎浩史,宇野泰広 .  カニクイザルの小腸に発現しているチトクロームP450タンパク質 .  第154回日本獣医学会学術集会 

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    開催年月日: 2012年9月

    記述言語:日本語   会議種別:口頭発表(一般)  

  • 中西康晴, 山下浩幸, 吉川剛, 鵜藤雅裕, 中村稚加, 宇野泰広 .  カンボジア産カニクイザルの小腸におけるチトクロームP450酵素活性 .  第39回日本毒性学会学術年会 

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    開催年月日: 2012年7月

    記述言語:日本語   会議種別:ポスター発表  

  • 藤原斉也,田坂克己,大浦華代子,上原正太郎,宇野泰広,今井輝子 .  ヒトおよびサル小腸の加水分解活性に関与するカルボキシルエステラーゼ分子種の同定 .  医療薬学フォーラム2012/第20回クリニカルファーマシーシンポジウム 

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    開催年月日: 2012年7月

    記述言語:日本語   会議種別:ポスター発表  

  • Hiroshi Yamazaki and Yasuhiro Uno .  Marmoset cytochrome P450: an extensive and unexploited field in drug metabolism studies .  第1回日本マーモセット研究会 

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    開催年月日: 2012年2月

    記述言語:日本語   会議種別:ポスター発表  

  • Tadashi Ishida, Sachiyo Sudou, Yuma Hatano, Yasuhiro Uno, Teruko Imai, and Masakiyo Hosokawa .  Transcriptional regulation and properties of monkey carboxylesterase 1 (CES1A) isozymes .  第26回日本薬物動態学会年会 

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    開催年月日: 2011年11月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno, Yasuharu Nakanishi, Ryota Ise, Hiroyuki Yamashita, Tsuyoshi Yoshikawa, Chika Nakamura, and Masahiro Utoh .  Hepatic expression of cytochromes P450 in cynomolgus monkeys from Cambodia, China, and Indonesia .  第26回日本薬物動態学会年会 

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    開催年月日: 2011年11月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuharu Nakanishi, Naoto Horai, Chika Nakamura, Masahiro Utoh, and Yasuhiro Uno .  Cytochrome P450 enzyme activities in the liver of juvenile cynomolgus monkeys .  第26回日本薬物動態学会年会 

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    開催年月日: 2011年11月

    記述言語:英語   会議種別:ポスター発表  

  • Hiroshi Yamazaki, Chie Emoto, Ryo Koizumi, Makiko Shimizu, Norie Murayama, Kazuhide Iwasaki, Masahiro Utoh, and Yasuhiro Uno .  Comparison of P450 2D and 3A-dependent drug oxidation activities in cynomolgus monkeys, microminipigs, and humans .  17th Annual ISSX North American Regional Meeting  国際会議

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    開催年月日: 2011年10月

    記述言語:英語   会議種別:ポスター発表  

  • 長田直樹、宇野泰広 .  機能喪失とCpGサイトの崩壊による旧世界ザルCYP1A2遺伝子の高い遺伝的多様性 .  日本遺伝学会第83回大会 

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    開催年月日: 2011年9月

    記述言語:日本語   会議種別:口頭発表(一般)  

  • 上原正太郎、村山典恵、山崎浩史、宇野泰広 .  カニクイザル肝臓におけるチトクロームP450タンパク質の発現 .  第152回日本獣医学会学術集会 

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    開催年月日: 2011年9月

    記述言語:日本語   会議種別:口頭発表(一般)  

  • 伊勢良太、吉川剛、山下浩幸、宇野泰広 .  カンボジア産カニクイザル肝臓のチトクロームP450 遺伝子発現解析 .  第38回日本トキシコロジー学会学術年会 

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    開催年月日: 2011年7月

    記述言語:日本語   会議種別:ポスター発表  

  • 中西康晴、山下浩幸、中村稚加、鵜藤雅裕、宇野泰広 .  カンボジア産カニクイザルの肝臓におけるチトクロームP450酵素活性 .  第38回日本トキシコロジー学会学術年会 

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    開催年月日: 2011年7月

    記述言語:日本語   会議種別:ポスター発表  

  • 福田浩司、小山周三、小原栄、有馬昭宏、宇野泰広、中間和浩、福崎好一郎 .  レボノルゲストレル投与したカニクイザルの腹腔鏡検査による排卵日の特定と卵巣への影響 .  第38回日本トキシコロジー学会学術年会 

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    開催年月日: 2011年7月

    記述言語:日本語   会議種別:ポスター発表  

  • 伊勢良太、近藤哲司、加藤寛人、今井統隆、秋山英雄、宇野泰広 .  カニクイザル肝臓におけるチトクロームP450遺伝子発現の年齢差 .  第38回日本トキシコロジー学会学術年会 

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    開催年月日: 2011年7月

    記述言語:日本語   会議種別:ポスター発表  

  • 小山周三、福田浩司、小原栄、有馬昭宏、宇野泰広、中間和浩、福崎好一郎 .  排卵同期化カニクイザルを用いた人工授精による妊娠個体作出の試み .  第38回日本トキシコロジー学会学術年会 

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    開催年月日: 2011年7月

    記述言語:日本語   会議種別:ポスター発表  

  • 田坂克己、大浦華代子、上原正太郎、宇野泰広、今井輝子 .  サルおよびヒトの2型カルボキシルエステラーゼ特性の比較 .  日本薬剤学会第26年会 

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    開催年月日: 2011年5月

    記述言語:日本語   会議種別:ポスター発表  

  • 宇野泰広 .  カニクイザルにおけるチトクロームP450の同定・解析 .  第1回CYP研究会  招待

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    開催年月日: 2010年11月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 宇野泰広 .  カニクイザルにおけるチトクロームP450の同定・解析 .  インハウスセミナー  招待

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    開催年月日: 2010年11月

    記述言語:日本語   会議種別:公開講演,セミナー,チュートリアル,講習,講義等  

  • 宇野泰広 .  カニクイザルにおけるチトクロームP450の同定・解析 .  第6回霊長類医科学フォーラム  招待

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    開催年月日: 2010年11月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • Yasuhiro Uno .  Identification and analysis of drug-metabolizing enzyme genes in cynomolgus monkey .  第25回日本薬物動態学会年会  招待

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    開催年月日: 2010年10月

    記述言語:英語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • Yasuhiro Uno, Shotaro Uehara, Norie Murayama, and Hiroshi Yamazaki .  Identification and analysis of CYP1D1 in cynomolgus monkey .  第25回日本薬物動態学会年会 

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    開催年月日: 2010年10月

    記述言語:英語   会議種別:ポスター発表  

  • Chie Emoto, Kazuhide Iwasaki, Ryo Koizumi, Masahiro Utoh, Norie Murayama, Yasuhiro Uno, and Hiroshi Yamazaki .  Species difference between cynomolgus monkey and human P450-dependent drug oxidations in liver microsomes .  第25回日本薬物動態学会年会 

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    開催年月日: 2010年10月

    記述言語:英語   会議種別:ポスター発表  

  • Kayoko Ohura, Katsumi Tasaka, Shotaro Uehara, Yasuhiro Uno, and Teruko Imai .  Comparison of enzyme characteristics between monkey and human carboxylesterase 2 .  第25回日本薬物動態学会年会 

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    開催年月日: 2010年10月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno .  Identification and analysis of cytochrome P450s in cynomolgus macaque .  International Primatological Society XXIII Congress Kyoto 2010  招待 国際会議

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    開催年月日: 2010年9月

    記述言語:英語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 小山周三、福田浩司、渡辺翔、松下聡紀、小原栄、宇野泰広、細井美彦 .  カニクイザルにおける着床前遺伝子診断を用いた効率的な個体作出の試み .  第37回日本トキシコロジー学会学術年会 

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    開催年月日: 2010年6月

    記述言語:日本語   会議種別:ポスター発表  

  • 小山周三、藤浪菜穂子、福田浩司、松下聡紀、渡辺翔、小原栄、宇野泰広、土屋英明、鳥居隆三、細井美彦 .  カニクイザル受精卵における着床前遺伝子診断用サンプル採取の検討 .  第57回日本実験動物学会総会 

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    開催年月日: 2010年5月

    記述言語:日本語   会議種別:ポスター発表  

  • 伊勢良太,宇野泰広 .  DNAマイクロアレイによるカニクイザルの遺伝子発現解析の有用性 .  第37回日本トキシコロジー学会学術年会  招待

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    開催年月日: 2010年3月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 古泉良、村山典恵、宇野泰広、清水万紀子、岩崎一秀、山崎浩史 .  ブフラロールの酸化的代謝に認められる特徴的なサルP450 2Dの酵素機能 .  第130回日本薬学会年会 

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    開催年月日: 2010年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 村山典恵、今村裕樹、大久保真穂、小野瀬優維、國兼絵里子、松本史彦、佐々木国玄、清水万紀子、山崎浩史、宇野泰広、鵜藤雅裕、岩崎一秀 .  ヒトと実験動物間で認められる薬物代謝酵素機能の種差 .  第130回日本薬学会年会 

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    開催年月日: 2010年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 細川正清,堀武志,須藤祥代,波多野由麻,倉林英次郎,宇野泰広,今井輝子 .  ヒトおよび実験動物カルボキシルエステラーゼのファミリー間での臓器差およびプロドラッグの代謝活性化の差異 .  第130回日本薬学会年会 

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    開催年月日: 2010年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 宇野泰広、上原正太郎、村山典恵、山崎浩史 .  カニクイザルCYP2G2の同定・解析 .  第150回日本獣医学会学術集会 

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    開催年月日: 2010年3月

    記述言語:日本語   会議種別:口頭発表(一般)  

  • 宇野泰広 .  カニクイザルにおけるチトクロームP450遺伝子の同定・解析 .  第130回日本薬学会年会  招待

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    開催年月日: 2010年3月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 細川正清,須藤祥代,堀武志,宇野泰広,今井輝子 .  サルおよびヒトカルボキシルエステラーゼの構造と機能の比較 .  第130回日本薬学会年会  招待

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    開催年月日: 2010年3月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 宇野泰広 .  サル類におけるチトクロームP450の解析-よりヒトに近いモデル動物作出に向けて- .  第29回日本臨床薬理学会年会  招待

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    開催年月日: 2009年12月

    記述言語:日本語   会議種別:公開講演,セミナー,チュートリアル,講習,講義等  

  • Yasuhiro Uno .  Identification and characterization of cytochrome P450 cDNAs in cynomolgus macaque; an implication for species difference between macaque and human in drug metabolism .  第24回日本薬物動態学会年会  招待

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    開催年月日: 2009年11月

    記述言語:英語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • Ryota Ise, Yasuhiro Uno, Hideo Akiyama, Satoshi Kondo, Hitoshi Nobumasa, and Ryoichi Nagata .  Development and evaluation of the custom-designed DNA microarray for cytochrome P450 in cynomolgus monkey .  第24回日本薬物動態学会年会 

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    開催年月日: 2009年11月

    記述言語:英語   会議種別:ポスター発表  

  • Shizuo Narimatsu, Takanori Naito, Takeshi Shimizudani, Nobumitsu Hanioka, Kazufumi Masuda, Takashi Katsu, Shigeru Yamano, Yasuhiro Uno, and Shinsaku Naito .  The roles of amino acid residues in the oxidation of substrates by primate CYP1A enzymes .  Symposium on Molecular Chirality 2009 

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    開催年月日: 2009年5月

    記述言語:日本語   会議種別:ポスター発表  

  • Ryota Ise, Yasuhiro Uno, Hideo Akiyama, Satoshi Kondo, Hitoshi Nobumasa, and Ryoichi Nagata .  Development of a DNA microarray to evaluate gene expression profiles of drug-metabolizing genes in the cynomolgus monkey .  Society of Toxicology’s 48th Annual Meeting  国際会議

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    開催年月日: 2009年3月

    記述言語:英語   会議種別:ポスター発表  

  • 須藤祥代、宇野泰広、今井輝子、細川正清 .  カニクイザルcarboxylesterase1(CES1)の発現調節機構 .  第129回日本薬学会年会 

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    開催年月日: 2009年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 上原正太郎,村山典恵,鵜藤雅裕,山崎浩史,宇野泰広 .  カニクイザルにおける新規CYP2Aの解析 .  BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会) 

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    開催年月日: 2008年12月

    記述言語:日本語   会議種別:ポスター発表  

  • 宇野泰広,上原正太郎,松下聡紀,小原栄,永田良一,福崎好一郎,鵜藤雅裕,村山典恵,山崎浩史 .  マカクザルにおける新規CYP2C遺伝子の解析 .  BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会) 

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    開催年月日: 2008年12月

    記述言語:日本語   会議種別:ポスター発表  

  • 松下聡紀,上原正太郎,小原栄,永田良一,福崎好一郎,鵜藤雅裕,宇野泰広 .  マカク属のサルCYP3Aにおける遺伝子多型の探索 .  BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会) 

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    開催年月日: 2008年12月

    記述言語:日本語   会議種別:ポスター発表  

  • Kayoko Ohura, Chie Okayama, Katsumi Tasaka, Shotaro Uehara, Yasuhiro Uno, Teruko Imai .  Cloning of monkey intestinal carboxylesterase and stable expression of its cDNA in HEL293 cells .  第23回日本薬物動態学会年会 

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    開催年月日: 2008年10月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno, Shotaro Uehara, Akinori Matsushita, Sakae Kohara, Ryoichi Nagata, Koichiro Fukuzaki, Masahiro Utoh, Norie Murayama, and Hiroshi Yamazaki .  Molecular characterization of novel CYP2C in macaques .  15th Annual ISSX North American Regional Meeting  国際会議

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    開催年月日: 2008年10月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno, Shotaro Uehara, Akinori Matsushita, Sakae Kohara, Ryoichi Nagata, Koichiro Fukuzaki, Masahiro Utoh, Norie Murayama, and Hiroshi Yamazaki .  Functional characterization of novel CYP2Cv4 in macaques .  第23回日本薬物動態学会年会 

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    開催年月日: 2008年10月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuharu Nakanishi, Akinori Matsushita, Kiyomi Matsuno, Hiroko Satoh, Masahiro Utoh, Chika Nakamura, and Yasuhiro Uno .  Distribution of CYP enzyme activities in the liver and small intestine of cynomolgus monkeys .  第23回日本薬物動態学会年会 

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    開催年月日: 2008年10月

    記述言語:英語   会議種別:ポスター発表  

  • 宇野泰広 .  サルにおけるチトクロームP450の解析-よりヒトに近いモデル動物作出に向けて- .  インハウスセミナー  招待

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    開催年月日: 2008年9月

    記述言語:日本語   会議種別:公開講演,セミナー,チュートリアル,講習,講義等  

  • 中島敏晶,大谷仁志,颯田葉子,宇野泰広,明里宏文,石田貴文,木村彰方 .  霊長類におけるToll-like receptor(TLR)関連遺伝子の分子進化と自然選択 .  第10回日本進化学会大会 

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    開催年月日: 2008年8月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 宇野泰広,上原正太郎,福崎好一郎,鵜藤雅裕 .  サル類におけるチトクロームP450の解析 .  第17回サル類疾病ワークショップ  招待

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    開催年月日: 2008年7月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 今井輝子,岡山知恵,大浦華代子,上原正太郎,宇野泰広 .  サル小腸における加水分解酵素の発現とその活性 .  日本薬剤学会第23年会 

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    開催年月日: 2008年5月

    記述言語:日本語   会議種別:ポスター発表  

  • 宇野泰広 .  カニクイザルCYP2C76の同定・解析 .  インハウスセミナー  招待

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    開催年月日: 2008年4月

    記述言語:日本語   会議種別:公開講演,セミナー,チュートリアル,講習,講義等  

  • 宇野泰広 .  カニクイザルCYP2C76遺伝子の同定・解析 .  第10回SNBL研究会  招待

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    開催年月日: 2008年2月

    記述言語:日本語   会議種別:公開講演,セミナー,チュートリアル,講習,講義等  

  • 宇野泰広,松下聡紀,小原栄,永田良一,鵜藤雅裕,福崎好一郎 .  アカゲザルにおける新規CYP2C遺伝子の検出および解析 .  第5回日本予防医学会学術総会 

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    開催年月日: 2007年11月

    記述言語:日本語   会議種別:ポスター発表  

  • Akinori Matsushita, Sakae Kohara, Masahiro Utoh, Ryoichi Nagata, Koichiro Fukuzaki, and Yasuhiro Uno .  Initial screening for CYP3A genetic polymorphisms in macaques .  8th International ISSX Meeting  国際会議

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    開催年月日: 2007年10月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno, Akinori Matsushita, Sakae Kohara, Ryoichi Nagata, Masahiro Utoh, and Koichiro Fukuzaki .  Identification and analysis of novel CYP2C in macaques .  8th International ISSX Meeting  国際会議

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    開催年月日: 2007年10月

    記述言語:英語   会議種別:ポスター発表  

  • 宇野泰広,松下聡紀,小原栄,永田良一,鵜藤雅裕,福崎好一郎 .  マカク新規CYP2C遺伝子の検出および解析 .  第144回日本獣医学会学術集会 

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    開催年月日: 2007年9月

    記述言語:日本語   会議種別:口頭発表(一般)  

  • Hideki Fujino, Yasuhiro Uno, Takayuki Kumano, Go Kito, Ryoichi Nagata, and Tetsuya Kamataki .  CYP2C76-mediated species difference in drug metabolism: A comparison of pitavastatin metabolism between monkeys and humans .  第21回日本薬物動態学会年会 

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    開催年月日: 2006年11月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno, Shinya Hosaka, Kiyomi Matsuno, Chika Nakamura, Go Kito, Tetsuya Kamataki, and Ryoichi Nagata .  Characterization of nine cytochrome P450 cDNAs in cynomolgus monkey .  第21回日本薬物動態学会年会 

     詳細を見る

    開催年月日: 2006年11月

    記述言語:英語   会議種別:ポスター発表  

  • 宇野泰広 .  よりヒトに近いモデル動物の作出にむけたサルCYP2C76の解析 .  第40回日本実験動物技術者協会総会  招待

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    開催年月日: 2006年10月

    記述言語:日本語   会議種別:公開講演,セミナー,チュートリアル,講習,講義等  

  • Yasuhiro Uno .  Identification and characterization of monkey-specific CYP2C76: Towards establishment of the better animal model .  16th International Symposium on Microsomes and Drug Oxidations  招待 国際会議

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    開催年月日: 2006年9月

    記述言語:英語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 中島敏晶、大谷仁志、颯田葉子、宇野泰広、明里宏文、石田貴文、木村彰方 .  霊長類におけるToll-like receptor(TLR)関連遺伝子の分子進化と自然選択 .  第15回日本組織適合性学会 

     詳細を見る

    開催年月日: 2006年9月

    記述言語:日本語   会議種別:ポスター発表  

  • 宇野泰広 .  サルCYP2C76の同定と解析-よりヒトに近いモデル動物の作出に向けて .  第10回薬物動態談話会セミナー  招待

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    開催年月日: 2006年8月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 鈴木穣,宇野泰広,中井謙太,菅野純夫 .  完全長cDNAを用いた種特異的転写産物/プロモーターの同定と解析 .  第79回日本薬理学会年会 

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    開催年月日: 2006年3月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • 宇野泰広,櫻庭大子,鬼頭剛,鎌滝哲也,永田良一 .  よりヒトに近いモデル動物の作出に向けたカニクイザルCYP2C76遺伝子多型の検出・解析 .  第126回日本薬学会年会 

     詳細を見る

    開催年月日: 2006年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 宇野泰広 .  カニクイザルCYP2C76遺伝子の同定・解析 .  SNBL研修セミナー  招待

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    開催年月日: 2006年1月

    記述言語:日本語   会議種別:公開講演,セミナー,チュートリアル,講習,講義等  

  • 宇野泰広 .  サル特異的CYP2C76の検出および機能解析(よりヒトに近いモデル動物の作出にむけて) .  日本薬物動態学会談話会特別例会  招待

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    開催年月日: 2005年11月

    記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

  • Yasuhiro Uno, Go Kito, Tetsuya Kamataki, and Ryoichi Nagata .  Characterization of newly identified CYP2C76 in cynomolgus monkey .  14th North American ISSX and 20th JSSX Joint Meeting  国際会議

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    開催年月日: 2005年10月

    記述言語:英語   会議種別:ポスター発表  

  • 宇野泰広,鬼頭剛,鎌滝哲也,永田良一 .  カニクイザル新規CYP2Cのゲノム構造 .  第125回日本薬学会年会 

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    開催年月日: 2005年3月

    記述言語:日本語   会議種別:ポスター発表  

  • Yasuhiro Uno, Go Kito, Tetsuya Kamataki, and Ryoichi Nagata .  Analysis of gene structure for novel CYP2C in cynomolgus monkey .  第27回日本分子生物学会 

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    開催年月日: 2004年12月

    記述言語:日本語   会議種別:ポスター発表  

  • 宇野泰広,鬼頭剛,鎌滝哲也,永田良一 .  カニクイザル新規CYP2Cのゲノム構造 .  第19回日本薬物動態学会 

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    開催年月日: 2004年11月

    記述言語:英語   会議種別:口頭発表(一般)  

  • 宇野泰広 .  カニクイザルCYP遺伝子の解析-薬物代謝の種差と性差の解明に向けて- .  SNBL研修セミナー  招待

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    開催年月日: 2004年5月

    記述言語:日本語   会議種別:公開講演,セミナー,チュートリアル,講習,講義等  

  • Yasuhiro Uno, Go Kito, Tetsuya Saito, Hiroshi Yamazaki, Tetsuya Kamataki, and Ryoichi Nagata .  Isolation and analysis of cynomolgus monkey CYP cDNAs .  Pharmaceutical Sciences World Congress 2004  国際会議

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    開催年月日: 2004年3月

    記述言語:英語   会議種別:ポスター発表  

  • 宇野泰広,鬼頭剛,斎藤鉄也,山崎浩史,鎌滝哲也,永田良一 .  カニクイザルCYP2C遺伝子の発現解析 .  第124回日本薬学会年会 

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    開催年月日: 2004年3月

    記述言語:日本語   会議種別:ポスター発表  

  • 宇野泰広,鬼頭剛,斎藤鉄也,山崎浩史,井ノ上逸郎,鎌滝哲也,永田良一 .  カニクイザル肝臓におけるCYPをコードするcDNAクローンの検出および解析 .  第26回日本分子生物学会 

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    開催年月日: 2003年12月

    記述言語:日本語   会議種別:ポスター発表  

  • 宇野泰広,鬼頭剛,斎藤鉄也,山崎浩史,鎌滝哲也,永田良一 .  ヒトCYP2Cと相同性の低いカニクイザルCYP2C .  第18回日本薬物動態学会 

     詳細を見る

    開催年月日: 2003年10月

    記述言語:英語   会議種別:口頭発表(一般)  

  • Yasuhiro Uno and F. Abel Ponce de Leon .  Chromosomal localization and gene expression analysis of chicken slow-type myosin binding protein C, nebulin, and phosphoglucomutase genes .  12th North American Colloquium on Animal Cytogenetics and Gene Mapping 

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    開催年月日: 2001年7月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno and F. Abel Ponce de Leon .  Isolation of chicken slow type myosin binding protein C (MyBP-C) gene and analysis of its expression pattern during muscle development .  87th Poultry Science Association Annual Meeting 

     詳細を見る

    開催年月日: 1998年8月

    記述言語:英語   会議種別:ポスター発表  

  • Yasuhiro Uno and F. Abel Ponce de Leon .  Identification of differentially expressed sequences in chicken skeletal muscle development .  87th Poultry Science Association Annual Meeting 

     詳細を見る

    開催年月日: 1998年8月

    記述言語:英語   会議種別:ポスター発表  

▼全件表示

知的財産権

  • 検出キット,及び検出方法

    宇野泰広,鵜藤雅裕,小原栄

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    出願番号:特願2015-136553  出願日:2015年7月

    特許番号/登録番号:6468555  登録日:2018年12月 

    出願国:国内  

  • サルの新規薬物代謝酵素

    宇野泰広,上原正太郎,鵜藤雅裕

     詳細を見る

    出願番号:特願2014-069496   出願日:2014年3月

    特許番号/登録番号:5782544  登録日:2015年6月 

    出願国:国内  

  • サルCYP2CおよびCYP2D遺伝子の多型の検出およびその利用

    宇野泰広,上原正太郎,松下聡紀,小原栄,鵜藤雅裕

     詳細を見る

    出願番号:特願2013-191104  出願日:2013年9月

    特許番号/登録番号:6342634  登録日:2018年4月 

    出願国:国内  

  • サル CYP3A 遺伝子多型の検出と利用

    宇野泰広,松下聡紀,鵜藤雅裕,福崎好一郎,永田良一

     詳細を見る

    出願番号:特願2009-71595  出願日:2009年3月

    特許番号/登録番号:5543120  登録日:2014年4月 

    出願国:国内  

  • サル薬物代謝酵素をコードする遺伝子の多型、およびその利用

    宇野泰広,永田良一,鬼頭剛,和泉博之

     詳細を見る

    出願番号:特願2006-553925  出願日:2006年1月

    特許番号/登録番号:5063117  登録日:2012年7月 

    出願国:国内  

  • サルの新規薬物代謝酵素、およびそれをコードする核酸

    宇野泰広,永田良一,鬼頭剛

     詳細を見る

    出願番号:特願2006-553924  出願日:2006年1月

    特許番号/登録番号:4972413  登録日:2012年3月 

    出願国:国内  

▼全件表示

受賞

  • 第25回日本薬物動態学会奨励賞

    2010年10月   日本薬物動態学会  

    宇野泰広

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    受賞区分:国内学会・会議・シンポジウム等の賞  受賞国:日本国

  • Graduate Student Research Certificate of Excellence

    1998年8月   Poultry Science Association  

    Yasuhiro Uno

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    受賞区分:国内学会・会議・シンポジウム等の賞  受賞国:アメリカ合衆国

  • University of Minnesota Animal Biotechnology Center Travel Award

    1998年7月   University of Minnesota  

    Yasuhiro Uno

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    受賞国:アメリカ合衆国

共同研究・競争的資金等の研究

  • 主な獣医動物種での薬物代謝研究の基盤構築を目指した責任酵素の網羅的な同定・解析

    2020年4月 - 2023年3月

    科学研究費補助金  基盤研究(C)

 

担当経験のある授業科目

  • 獣医薬理学B

    2020年11月
    -
    現在
    機関名:鹿児島大学

     詳細を見る

    科目区分:学部専門科目 

  • 基礎獣医学特別講義(分子薬理毒性学特別講義)

    2020年10月
    -
    現在
    機関名:鹿児島大学

     詳細を見る

    科目区分:大学院専門科目 

  • 獣医毒性学B

    2020年6月
    -
    現在
    機関名:鹿児島大学

     詳細を見る

    科目区分:学部専門科目 

  • 獣医薬理毒性学実習

    2020年4月
    -
    現在
    機関名:鹿児島大学

     詳細を見る

    科目区分:学部専門科目 

  • 獣医薬理学D

    2019年12月
    -
    現在
    機関名:鹿児島大学

     詳細を見る

    科目区分:学部専門科目