Aromatase Reaction of Suicide Substrates of Aromatase and Their Inactivation Mechanisms.

芳香酶自杀底物的芳香酶反应及其失活机制。

• 批准号: 09672246
• 负责人: NUMAZAWA Mitsuteru
• 金额: $ 1.22万
• 依托单位: Tohoku College of Pharmacy
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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-09672246/
• 关键词: Aromatase Inhibitor; 6-Alkyl steroid; 3-Deoxy steroid; Aromatization; 19-Oxygenation; Gas chromatography; Mass spectrometry; Breast cancer; アロマターゼ; 阻害剤; 6-アルキルアンドロゲン; 芳香核化反応; 乳ガン化学療法剤

We have obtained the following results by the project carried out at 1997 and 19981. Kinetics of aromatization of suicide substrates of aromatase, DELTA^<1_>, DELTA^<6_> and DELTA^<1.6_> androstenediones (AD) and their 6-alkyl analogs, with human placental aromatase was analyzed by GC-MS.DELTA^<1_>-AD was a good substrate for the enzyme and its aromatization rate was comparable with that of AD, in contrast, DELTA^<6_> and DELTA^<1.6_>-AD's were the poor substrates. The stereochemistry and length of the 6-alkyl group affected the aromatization to a different extent in each series. There was no direct correlation etween the K_i and K_m values for the inhibitors but the aromatization of DELTA^<1_> and DELTA^<1.6_> AD analogs correlated well to their inactivation reactions.2. [19^<_3>H, 4-^<_14>C]-DELTA^<1_>-AD synthesized chemically, was aromatized accompanying with the release of ^3H_20 and ^3HCOOH, indicating that DELTA^1-AD aromatizes via the same mechanism involved in the AD aromatization.3. Structure-activity relationships of 6-phenylaliphatic AD's as well as their DELTA^<1_>, DELTA^<6_>, and DELTA^<1.6_> analogs as aromatase inhibitors inactivators showed new aspects for a binding manner of the inhibitors to the active site.4. Three regioisomers of AD having a double-bond in A, B-ring system as well as 3-deoxy steroids was oxygenated at C-19 position with aromatase.5.19-Oxygenated 4beta, 5beta-epoxy AD's were aromatized to estrone with aromatiase.

我们通过1997年和1998年实施的项目取得了以下成果1。用GC-MS分析了芳香化酶的自杀底物DELTA^-AD<1_>、DELTA^-AD<6_>和DELTA <1.6_>^-AD及其6-烷基类似物与人胎盘芳香化酶的芳构化反应动力学，DELTA^<1_>-AD是该酶的良好底物，其芳构化反应速率与AD相当，DELTA^-AD<6_>和DELTA^<1.6_>-AD's是较差的底物。6-烷基的立体化学和长度对芳构化反应有不同程度的影响。抑制剂的K_i和K_m值之间没有直接的相关性，但DELTA_i<1_>和DELTA_i <1.6_>AD类似物的芳构化与其失活反应有很好的相关性. [19化学合成的Δ <_3>H，4-<_14>C]-Δ ^<1_>-AD发生芳构化反应，并伴随着^3H_20和^3HCOOH的释放，表明Δ ^-AD的芳构化反应与AD的芳构化反应机理相同。6-苯基脂肪族AD及其类似物<1_><6_><1.6_>作为芳香酶抑制剂失活剂的构效关系显示了抑制剂与活性位点结合方式的新方面。用芳香化酶将AD的三个A、B环上有双键的区域异构体以及3-脱氧甾体在C-19位氧化。5.19-氧化的4 β，5 β-环氧AD用芳香化酶芳构化为雌酮。

项目成果

Mitsuteru Numazawa, Satoshi Yamaguchi: "6-Phenylaliphatic-substituted Androst-4-ene-3,17-diones as Aromatase Inhibitors : Structure-activity Relationships." J.Steroid Biochem.Molec.Biol.67. 41-48 (1998)

Mitsuteru Numazawa、Satoshi Yamaguchi：“6-苯基脂肪族取代的 Androst-4-ene-3,17-二酮作为芳香酶抑制剂：结构-活性关系。”

Mitsuteru Numazawa, Keiko Yamada: "Reaction of Androst-5-en-17-one with Hypobromous Acid and Its Use for Synthesis of 19-Oxygenated 5-ene and 4-en-6-one Steroids." Steroids. 63. 62-69 (1998)

Mitsuteru Numazawa、Keiko Yamada：“Androst-5-en-17-one 与次溴酸的反应及其用于合成 19-氧化 5-ene 和 4-en-6-one 类固醇的用途。”

Masao Nagaoka, Makoto Morio and Mitsuteru Numazawa: "Synthesis of Deuterium-Labeled Androst-5-ene-17beta, 19-diol and Its 4-Ene Isomer as Internal Standards for Determination of the of Aromatase Inhibitors Using GC-MS." Chem.Pham.Bull.47. 263-266 (1999)

Masao Nagaoka、Makoto Morio 和 Mitsuteru Numazawa：“合成氘标记的 Androst-5-ene-17beta、19-二醇及其 4-Ene 异构体，作为使用 GC-MS 测定芳香酶抑制剂的内标。”

沼沢光輝: "Enzymic aromatization of 6-alkyl-substituted androgens, potent competitive and mechanism-based inhibitors of aromatase" Biochemical Journal. 329. 151-156 (1998)

Mitsuteru Numazawa：“6-烷基取代雄激素的酶芳香化，芳香酶的有效竞争性和基于机制的抑制剂”生物化学杂志 329. 151-156 (1998)。

沼沢光輝: "Aromatization of 6-Alkyl-substituted Androgens, Potent Competitive and Mechanism-based Inhibitors of Aromatase" Biochem.J.329. 151-156 (1998)

Mitsuteru Numazawa：“6-烷基取代雄激素的芳香化，芳香酶的有效竞争性和基于机制的抑制剂”Biochem.J.329（1998）。