Studies on the prediction of fetal toxicity caused by drugs

药物引起的胎儿毒性预测研究

• 批准号: 09470522
• 负责人: KITADA Mitsukazu
• 金额: $ 6.78万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470522/
• 关键词: CYP; CYP3A7; steroid metabolism; drug metabolism; fetal toxicity; CYP3A

Human CYP3A4, CYP3A7 and their chimera CYPs expressed in TN5 or SF9 cells using baculovirus were used for the characterization of human fetal CYP3A7 in steroid metabolism, especially dehydroepiandrosterone (DHEA) and DHEA 3-sulfate (DHEA-3S).(1) Expression levels of CYP3A enzymes and their chimera enzymes in insect cells and the stability of expressed proteins were different among these GYP enzymes. Exogenous NADPH-cytochrome P450 reductase and cytochrome b5 were essential to obtain the maximal activities for 16 - hydtoxylations of DHEA.and DHEA-3S.(2) It was suggested that the amino acid residues from 210 to 279 in CYP3A7 were important to hydroxylate DHEA and DHEA-3S at 16 position.(3) Computer model of CYP3A7 indicated that the protein was stabilized to bind with DHEA and DHEA-3S to form the hydrogen bond at Asp214 with hydroxyl group (DHEA) or -OSO3H (DHEA-3S) at Asp2l4 (oxygen of -CONH2 in Asp).(4) From the kinetic studies, it was suggested that the region from 40 to 109 amino acids residues in CYP3A7 might be important to interact with NADPH-cytochrome P450 reductase and the region from 210 to 279 amino acid residues might be interact with alpha-naphthoflavone.

利用杆状病毒在TN5或SF9细胞中表达的人CYP3A4、CYP3A7及其嵌合体CYPs，用于表征人胎儿CYP3A7在类固醇代谢中的作用，特别是脱氢表雄酮（DHEA）和硫酸脱氢表雄酮（DHEA- 3s）。(1)这些GYP酶在昆虫细胞中CYP3A酶及其嵌合体酶的表达水平和表达蛋白的稳定性存在差异。外源性nadph -细胞色素P450还原酶和细胞色素b5是获得DHEA 16 -羟化活性的必要条件。和DHEA-3S。(2) CYP3A7中210 ~ 279个氨基酸残基对16位羟基化DHEA和DHEA- 3s具有重要意义。(3) CYP3A7的计算机模型表明，该蛋白稳定与DHEA和DHEA- 3s结合，在Asp214处与羟基（DHEA）或-OSO3H (DHEA- 3s) （Asp中-CONH2的氧）形成氢键。(4)动力学研究表明，CYP3A7中40 ~ 109个氨基酸残基可能与nadph -细胞色素P450还原酶相互作用，210 ~ 279个氨基酸残基可能与α -萘黄酮相互作用。

项目成果

Ohmori S,Nakasa H,Asanome K,Kurose Y,Ishii I,Hosokawa M and Kitada M: "Differential catalytic properties in metabolism of endogenous and exogenous substrates among CYP3A enzymes expressed in COS-7 cells." Biochim Biophys Acta. 1380. 297-304 (1998)

Ohmori S、Nakasa H、Asanome K、Kurose Y、Ishii I、Hosokawa M 和 Kitada M：“COS-7 细胞中表达的 CYP3A 酶内源性和外源性底物代谢的不同催化特性。”

Chmoris, Nakasa H, Asanoue, K.Kurose Y, Ishii I, Hosokawa M, Kitada M: "Differential corolytic properties in metabolism of endogenous and exaganous substrate among CYP3A enzymes expressed in Cas celle" Biochim.Biophys.Acta. 1380. 294-304 (1998)

Chmoris、Nakasa H、Asanoue、K.Kurose Y、Ishii I、Hosokawa M、Kitada M：“Cas celle 中表达的 CYP3A 酶内源性和外源性底物代谢中的不同 corolytic 特性”Biochim.Biophys.Acta。

Ohmori S,Fujiki N,Nakasa H,Nakamura H,Ishii I,Itahashi K,Kitada M: "Steroid Hydroxylation by human fetal CYP3A7 and human NADPH-cytochrome P450 expressed in insect cells using baculovirus" Res.Commun Molec Pathol Pharmacol. 100. 15-28 (1998)

Ohmori S、Fujiki N、Nakasa H、Nakamura H、Ishii I、Itahashi K、Kitada M：“使用杆状病毒在昆虫细胞中表达的人胎儿 CYP3A7 和人 NADPH-细胞色素 P450 进行类固醇羟化”Res.Commun Molec Pathol Pharmacol。

Ohmori S,Fujiki N,Nakasa H,Nakamura H,Ishii I, Itahashi K and Kitada M: "Steroid hydroxylation by human fetal CYP3A7 and human NADPH-cytochrome P450 reductase coexpressed in insect cells using baculovirus." Res Commun Molec Pathol Pharmacol. 100. 15-28 (1

Ohmori S、Fujiki N、Nakasa H、Nakamura H、Ishii I、Itahashi K 和 Kitada M：“使用杆状病毒在昆虫细胞中共表达的人胎儿 CYP3A7 和人 NADPH-细胞色素 P450 还原酶的类固醇羟基化。”

Ohmori S,Nakasa H,Asanoma K,Kurose Y,Ishii I,Hisakawa M,Kitada M: "Differential cadalytic properties in metabolism of endgenous and exogenous substrades among CYP3A enzymes expressed in CDS-cells" Biochim Biophys Acta. 1380. 297-304 (1998)

Ohmori S、Nakasa H、Asanoma K、Kurose Y、Ishii I、Hisakawa M、Kitada M：“CDS 细胞中表达的 CYP3A 酶中内源性和外源性底物代谢的不同催化特性”Biochim Biophys Acta。