Developmental study for effective high-through-put screening system for new drug registration

新药注册高效高通量筛选系统的开发研究

• 批准号: 11557175
• 负责人: KAMATAKI Tetsuya
• 金额: $ 5.25万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557175/
• 关键词: P450; HIGH THROUGH PUT; PLATE READER; INHIBITION; DRUG METANBOISM; DRUG INTERACTION; RECOMBINANT; DRUG DEVELOPMENT; ハイスループットシステム; チトクロームP450; CYP3A4; 大腸菌発現系; チトクロームb_5; 薬物間相互作用; 水溶性ポリマー

The role of human cytochrome P450 (CYP) in the metabolic activation of N-alkylnitrosamines was examined by Ames test using genetically engineered Salmonella typhimurium (S. typhimurium)YG7108 cells expressing each form of human CYP together with human NADPH-cytochrome P450 reductase (OR). The relationship between the structure of N-alkylnitrosamines and CYP form(s) involved in the activation was evaluated. Eleven strains of S. typhimurium YG7108 cells expressing each form of CYP (CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4 or CYP3A5) were employed. Eight N-alkylnitrosamines These N-alkylnitrosamines were mainly activated by CYP2E1, CYP2A6 and CYP1A1. N-alkylnitrosamines with relatively short alkyl chains such as NDMA and NMEA were primarily activated by CYP2E1 as judged by mutagen-producing capacity. With the increase of the number of the carbon atoms of the alkyl chains, the contribution of CYP2A6 increased. CYP2A6 played major roles in the activation of NDEA, NDPA, NMPA, NMBA and NEBA. Interestingly, CYP1A1 became a molecular form of CYP playing a major role in the metabolic activation of NDBA.

通过艾姆斯试验，使用基因工程鼠伤寒沙门氏菌 (S.typhimurium)YG7108 细胞表达每种形式的人 CYP 以及人 NADPH-细胞色素 P450 还原酶 (OR)，检查人细胞色素 P450 (CYP) 在 N-烷基亚硝胺代谢激活中的作用。评估了 N-烷基亚硝胺的结构和参与活化的 CYP 形式之间的关系。使用表达每种形式的 CYP (CYP1A1、CYP1A2、CYP1B1、CYP2A6、CYP2C8、CYP2C9、CYP2C19、CYP2D6、CYP2E1、CYP3A4 或 CYP3A5)的 11 株鼠伤寒沙门氏菌 YG7108 细胞株。八种N-烷基亚硝胺 这些N-烷基亚硝胺主要由CYP2E1、CYP2A6和CYP1A1激活。从诱变剂产生能力来看，烷基链较短的N-烷基亚硝胺（例如NDMA和NMEA）主要被CYP2E1激活。随着烷基链碳原子数的增加，CYP2A6的贡献增加。 CYP2A6 在 NDEA、NDPA、NMPA、NMBA 和 NEBA 的激活中起主要作用。有趣的是，CYP1A1 成为 CYP 的分子形式，在 NDBA 的代谢激活中发挥重要作用。

项目成果

鎌滝哲也: "薬物療法学"南山堂 (in press).

Tetsuya Kamataki：“药物治疗学”Nanzando（正在出版）。

Noritaka Ariyoshi et al.: "Handbook of Experimental Pharmacology"Springer-VerlagBerlin. (2001)

Noritaka Ariyoshi 等人：“实验药理学手册”Springer-VerlagBerlin。

Ken-ichi Fujita, Kazuo Nakayama, Yoshiyuki Yamazaki, Kazuhiro Tsuruma, Masami Yamada, Takehiko Nohmi, Tetsuya Kamataki: "Construction of Salmonella typphimurium YG7108 stains each co-expressing a form of human cytochrome P450 with NADPH-cytochrime P450 re

Ken-ichi Fujita、Kazuo Nakayama、Yoshiyuki Yamazaki、Kazuhiro Tsuruma、Masami Yamada、Takehiko Nohmi、Tetsuya Kamataki：“构建鼠伤寒沙门氏菌 YG7108 染色剂，每种染色剂与 NADPH-细胞色素 P450 共表达一种形式的人细胞色素 P450

Noritaka Ariyoshi et al.: "Studies on the genetic polymorphism of CYP2A6."Drug Metabolism and Disposition. 15. 57-61 (2000)

Noritaka Ariyoshi 等人：“CYP2A6 遗传多态性的研究”。药物代谢和处置。

Ken-ichi Fujita et al.: "Establishment of Salmonella strain expressing catalytically active human UDP-glucuronosyltransferase 1A1 (UGT1A1)"Life Science. 66. 1955-1967 (2000)

Ken-ichi Fujita 等人：“表达催化活性人 UDP-葡萄糖醛酸基转移酶 1A1 (UGT1A1) 的沙门氏菌菌株的建立”生命科学。