{"created":"2023-07-27T06:58:42.588361+00:00","id":57636,"links":{},"metadata":{"_buckets":{"deposit":"22f54dd9-62bf-4393-980f-4132ce45f2f2"},"_deposit":{"created_by":18,"id":"57636","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"57636"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00057636","sets":["2812:2813:2833"]},"author_link":["26718","95962"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2002-03-25","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"2p.","bibliographicVolumeNumber":"1999 – 2000","bibliographic_titles":[{"bibliographic_title":"平成12(2000)年度 科学研究費補助金 基盤研究(B) 研究成果報告書概要"},{"bibliographic_title":"2000 Fiscal Year Final Research Summary","bibliographic_titleLang":"en"}]}]},"item_9_creator_33":{"attribute_name":"著者別表示","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{}],"nameIdentifiers":[{},{}]}]},"item_9_description_21":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"実験動物との種差の問題から、ヒト由来の組織、培養細胞又はヒト酵素遺伝子発現系を用いてヒトにおける代謝反応を明らかにすることが必要となっている。第1相の薬物代謝反応において大きな役割を果たすチトクロムP450酵素(以下CYP又はP450)を中心にいくつかのヒトの薬物代謝酵素が試薬として製造あるいは輸入販売されている。最近市販されているP450関連試薬では、P450だけでなく、NADPHからの電子伝達に必須なNADPH-P450還元酵素(NPR)を同時に発現させた、いわゆる共発現系のミクロゾームが主流になっているが、各種発現系の選択とその結果の解釈に議論がなされている。我々は、ヒト肝に存在する薬物代謝型の主要分子種であるP450についてNPRとの共発現pCWュプラスミドを大腸菌に導入し、その膜画分の調製した。さらに大腸菌膜を用いたP450共発現系にさらにNPRやチトクロームb_5(b_5)を添加して触媒活性の至適条件を検索し、市販のリンパ芽球、バキュロウイルス発現系やヒト肝ミクロゾームの薬物酸化酵素活性と比較した。\n発現機構が未解明である薬物相互作用の一つにフェニトインとテガフールがある。抗てんかん薬フェニトインと抗癌剤テガフールを併用することによりフェニトインの血中濃度が上昇し、意識障害などのフェニトイン中毒と思われる臨床症状が現れることが報告されている。そこで本研究では、これらの薬物相互作用の発現機序を解明するために、ヒト肝P450によるテガフールとフェニトインの代謝について検討を行った。ヒト肝ミクロソームによる5-FU生成は、主にCYP1A2、CYP2A6およびCYP2C8によって触媒され、これらのP450の寄与の程度はヒト肝P450の組成比に依存して個人差が認められることが示された。ヒト肝において、定常血中濃度(100μM)付近のテガフールからの5-FU生成は主にミクロソーム画分の酵素(P450)によって触媒され、テガフール濃度が高い場合にはサイトゾル画分のチミジンホスホリラーゼにより触媒されることを示した。ヒト肝ミクロソームによるフェニトイン酸化活性は肝サイトゾルの添加によって増加し、サイトゾル存在下の3′,4′-diHPPH生成活性は主代謝物である4′-HPPH生成活性と同レベルであった。4′-HPPH生成には主にCYP2C9、一部CYP2C19が関与した。一方、3′,4′-diHPPH生成には主にCYP2C9、CYP2C19およびCYP3A4が関与しており、これらのP450の寄与の程度はヒト肝P450の組成比に依存して個人差が認められることを明らかにした。","subitem_description_type":"Abstract"},{"subitem_description":"Drug oxidation activities of twelve recombinant human cytochrome P450 (P450) coexpressed with human NADPH-P450 reductase (NPR) in bacterial membranes (P450/NPR membranes) were determined and compared with those of other recombinant systems and of human liver microsomes. Addition of exogenous membrane-bound NPR to the P450/NPR membranes enhanced the catalytic activities of CYP2C8, CYP2C9, CYP2C19, CYP3A4, and CYP3A5 ; however, enhancement of activities of CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2B6, CYP2D6, and CYP2E1 in membranes was not observed after the addition of NPR in 4-molar excess to each P450. Exogenous purified human cytochrome b_5 (b_5) further enhanced catalytic activities of CYP2A6, CYP2B6, CYP2C8, CYP2E1, CYP3A4, and CYP3A5/NPR membranes. Catalytic activities of CYP2C9 and CYP2C19 were enhanced by addition of b_5 in reconstitution systems but not in the P450/NPR membranes. Apo b_5 (devoid of heme) enhanced catalytic activities when added to the both systems, except for CYP2E1/NPR membranes and the reconstituted systems containing purified CYP2C8 or CYP2E1 in comparison with b_5. Catalytic activities in P450/NPR membranes plus b_5 systems were roughly similar to those measured with microsomes of insect cells coexpressing P450 with NPR (and b_5) and/or of human liver microsomes, based on equivalent P450 contents. These results suggest that interactions of P450 and NPR coexpressed in membranes and reconstituted systems appear to be different in some human CYP2 family enzymes, possibly due to a conformational role of b_5. P450/NPR membrane systems containing b_5 are useful models for prediction of the rates for liver microsomal P450-dependent drug oxidations.","subitem_description_type":"Abstract"}]},"item_9_description_22":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"研究課題/領域番号:11557191, 研究期間(年度):1999 – 2000","subitem_description_type":"Other"},{"subitem_description":"出典：「ヒト薬物代謝酵素を応用した化学物質の体内動態予測の基盤に関する研究」研究成果報告書　課題番号11557191\n（KAKEN：科学研究費助成事業データベース（国立情報学研究所））\n（https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-11557191/115571912000kenkyu_seika_hokoku_gaiyo/)を加工して作成","subitem_description_type":"Other"}]},"item_9_description_5":{"attribute_name":"提供者所属","attribute_value_mlt":[{"subitem_description":"金沢大学 薬学部","subitem_description_type":"Other"}]},"item_9_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00063906","subitem_identifier_reg_type":"JaLC"}]},"item_9_relation_28":{"attribute_name":"関連URI","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/search/?kw=30191274"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/search/?kw=30191274","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-11557191/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-11557191/","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-11557191/115571912000kenkyu_seika_hokoku_gaiyo/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-11557191/115571912000kenkyu_seika_hokoku_gaiyo/","subitem_relation_type_select":"URI"}}]},"item_9_version_type_25":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_ab4af688f83e57aa","subitem_version_type":"AM"}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2021-10-15"}],"displaytype":"detail","filename":"PH-PR-YAMAZAKI-H-kaken 2002-2p.pdf","filesize":[{"value":"106.3 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"PH-PR-YAMAZAKI-H-kaken 2002-2p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/57636/files/PH-PR-YAMAZAKI-H-kaken 2002-2p.pdf"},"version_id":"61dea688-206e-425d-b54c-bef044835b55"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"ヒト薬物代謝酵素を応用した化学物質の体内動態予測の基盤に関する研究","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"ヒト薬物代謝酵素を応用した化学物質の体内動態予測の基盤に関する研究"},{"subitem_title":"Prediction of drug disposition by human drug metabolizing enzymes","subitem_title_language":"en"}]},"item_type_id":"9","owner":"18","path":["2833"],"pubdate":{"attribute_name":"公開日","attribute_value":"2021-10-15"},"publish_date":"2021-10-15","publish_status":"0","recid":"57636","relation_version_is_last":true,"title":["ヒト薬物代謝酵素を応用した化学物質の体内動態予測の基盤に関する研究"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-07-27T14:43:20.310864+00:00"}