Design and Synthesis of New Sensitive Dienophiles, and Application to the Quantitative Analysis of Biologically Active Conjugated Dienes

新型敏感亲双烯体的设计与合成及其在生物活性共轭二烯定量分析中的应用

• 批准号: 05671741
• 负责人: SHIMIZU Masato
• 金额: $ 1.15万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671741/
• 关键词: fluorescent dienophile; fluorometric assay; conjugated dienes; cycloaddition reaction; vitamin D metabolites; retinoic asid; microcystin; chemiluminescence; 蛍光ラベル化; 付加反応

1. Synthesis of highly sensitive dienophilesFluorescent or chemiluminescent dienophiles targeting conjugated diene were designed and synthesized. A highly sensitive chemiluminescent dienophile, in which a chemiluminescent chromophore (acridinium ester group) and a reactive 1,2,4-triazoline-3,5-dione group were connected, was newly synthesized.2.A fluorescent dienophile (DMEQ-TAD) was applied to the fluorometric analysis of vitamin D,vitamin A and microcystins.(1) Fluorometric analysis of plasma vitamin D metabolites was examined and new accurate methods for assaying 25-OHD_3,24,25-(OH)_2D_3 and an active hormone 1,25-(OH)_2D_3 in human plasma were successfully developed.(2) Reactions of DMEQ-TED with retinoic acid and its geometrical isomers having a pentaene system were examined. Except for the 9Z-isomer, reactions gave the mono-adducts in high selectivity while the 9Z-isomer gave only the bis-adducts indicating that DMEQ-TAD might be useful as a fluorescence labeling for retinoic acids in biological fluid.(3) DMEQ-TAD reacted with microcystins, a cyclic heptaptide toxin from blue-green algae, at the conjugated diene part to yield the cycloadducts. The fluorometric method for assaying enviromental microcystins is under investigation.

1. 高灵敏度亲二烯试剂的合成设计并合成了针对共轭二烯的荧光或化学发光亲二烯试剂。新合成了一种由化学发光发色团（吖啶酯基）和反应性1,2,4-三唑啉-3,5-二酮基团连接而成的高度敏感的化学发光亲二烯亲和物。采用荧光亲二烯试剂（DMEQ-TAD）对维生素D、维生素A和微囊藻毒素进行荧光分析。(1)研究了血浆维生素D代谢物的荧光分析方法，成功地建立了测定人血浆中25- ohd_3、24,25-(OH)_2D_3和活性激素1,25-(OH)_2D_3的新方法。(2)考察了DMEQ-TED与维甲酸及其具有五烯体系的几何异构体的反应。除9z -异构体外，反应对单加合物具有高选择性，而9z -异构体仅对双加合物具有高选择性，这表明DMEQ-TAD可作为生物液中维甲酸的荧光标记物。(3) DMEQ-TAD与来自蓝绿藻的环七肽毒素微囊藻毒素在共轭二烯部分反应生成环加合物。荧光法测定环境微囊藻毒素的方法正在研究中。

项目成果

清水正人: "共役ジエンを標的とする新蛍光標識試薬の開発ならびに同試薬のビタミンDおよびA代謝物定量への応用" ビタミン. 68. 15-30 (1994)

Masato Shimizu：“针对共轭二烯的新型荧光标记试剂的开发及其在定量维生素 D 和 A 代谢物中的应用”维生素。 68. 15-30 (1994)

K.Yamanoto, J.Takahashi, K.Hamano, S.Yamada, K.Yamaguchi, H.Deluca: "Stereoselective Syntheses of (22R) -and (22S) -22-Methy1-1alpha, 25-dihydroxyvitamin D_3 : Active Vitamin D_3 Analogs with Rectricted Side Chain Conformation" J.Org.Chem. 58. 2530-2537 (

K.Yamanoto、J.Takahashi、K.Hamano、S.Yamada、K.Yamaguchi、H.Deluca：“(22R)-和(22S)-22-Methy1-1alpha、25-二羟基维生素 D_3 的立体选择性合成：活性维生素

G.Ying, M.Shimizu, S.Yamada, Y.Ozaki, T.Aso: "The Effects of Chemotherapy Including Cisplatin on Vitamin D Metabolism" Endocrine J.40. 737-742 (1993)

G.Ying、M.Shimizu、S.Yamada、Y.Ozaki、T.Aso：“包括顺铂在内的化疗对维生素 D 代谢的影响”内分泌 J.40。

M.Shimizu, T.Yamasaki, S.Yamada: "Fluorescence-Labeling Reaction of Vitamin D Meta-bolites and Analogs with Fluorescent 1,2,4-Triazoline-3,5-dione (DMEQ-TAD)" BioMed.Chem.Lett.9. 1809-1814 (1993)

M.Shimizu、T.Yamasaki、S.Yamada：“维生素 D 代谢物和类似物与荧光 1,2,4-三唑啉-3,5-二酮 (DMEQ-TAD) 的荧光标记反应”BioMed.Chem。

M.Shimizu, K.Yaguchi, Y.Iwasaki, S.Yamada: "Reaction of Geometrical Isomers of Retinoic Acid with 1,2,4-Triazoline-3,5-dione Having Fluorescent Chromophore" Tetrahedron Lett.35. 1917-1920 (1994)

M.Shimizu、K.Yaguchi、Y.Iwasaki、S.Yamada：“视黄酸几何异构体与具有荧光发色团的 1,2,4-三唑啉-3,5-二酮的反应”四面体 Lett.35。