A Study on Simple and Highly Sensitive Analytical Methods of Pteridines Aimed to be Used for the Detection of Virus Infection

用于病毒感染检测的蝶啶简单、高灵敏度分析方法的研究

• 批准号: 09640722
• 负责人: SUGIMOTO Takashi
• 金额: $ 1.98万
• 依托单位: NAGOYA UNIVERSITY
• 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-09640722/
• 关键词: enyme linked immunosorbent assay; neopterin; biopterin; arylation; arenediazonium salts; 1.3-dimethyllumazine-6-triflate; nucleophilic substitution with enolates; ネオプリテリン; エノラートとの求核置換反応; 位置選択的導入

We investigated enzyme linked immunosorbent assay for neopterin and biopterin by using polystyrene microtitre plate with 96 wells. ELISA was performed by adding the substances to the well with fixed hapten as follows : incubation mixture of antibody and hapten ; goat anti-rabbit IgG ; peroxidase antiperoxidase complex ; o-phenylendiamine and hydrogen peroxide ; sulfuric acid. The absorbance at 490nm was measured on a immunoreader. We have investigated a simpler assay method by employing horseraddish peroxidase-goat antirabbit IgG conjugate in place of goat anti rabbit IgG and peroxidase anti-peroxidase complex in the above procedure. The present two methods exhibited the almost same sensitivity (l-25pmol), reliability, specificity (the two haptens were almost completely distingushed).We also studied new methods to introduce carbon substituents into 6,7-unsubstituted pteridines. 1,3-Dimethyllumazine 5-oxide and arene diazonium salts gave various 6-arylation products, regioselectively. 1,3-Dimethyllumazine gave a mixture of 6- and 7-arylation products. 1,3-Dimethyllumazine 5-oxide and trifluoromethanesulfonic anhydride gave 1,3-dimethyllumazine-6-triflate, which gave various 6-substituted derivatives on reaction with enolates.

采用96威尔斯聚苯乙烯微量滴定板，建立了新蝶呤和生物蝶呤的酶联免疫吸附测定方法。通过向具有固定半抗原的孔中加入如下物质进行ELISA：抗体和半抗原的孵育混合物;山羊抗兔IgG ;过氧化物酶抗过氧化物酶复合物;邻苯二胺和过氧化氢;硫酸。在免疫读数器上测量490 nm处的吸光度。本文用辣根过氧化物酶-羊抗兔IgG偶联物代替羊抗兔IgG和过氧化物酶-抗过氧化物酶复合物，研究了一种简便的测定方法。本研究还探索了在6，7-未取代蝶啶类化合物上引入碳取代基的新方法。1，3-二甲基鲁马嗪5-氧化物和芳烃重氮盐区域选择性地得到了不同的6-芳基化产物。1，3-二甲基鲁马嗪得到6-和7-芳基化产物的混合物。1，3-二甲基二甲嗪-5-氧化物和三氟甲磺酸酐反应得到1，3-二甲基二甲嗪-6-三氟甲磺酸酯，后者与烯醇化物反应得到各种6-取代衍生物。

项目成果

村田静昭: "preparation ant synthe tie appLcation of highyg teachive attigluoremetha-nesulfonylcxy lumigine Anovel methat for 6-substituted lumazines" Pteridines. 8巻. 1-9 (1997)

Shizuaki Murata：“用于 6-取代 2-四氢呋喃的高浓度教学 attigluoremetha-nesulfonylcxy lumigine Anovel methat 的制备和合成应用”，蝶啶，第 8 卷，1-9 (1997)。

杉本隆: "A simple procedure to intrcdue an arylgroup into the ptetidine ring by areedingonium salts in on alkaline spltion" chemistry and liologg of pteridines and folates 1997 Eds Pfleideier & Rokos,Blackwell Science,Beslin. 69-72 (1997)

Takashi Sugimoto：“通过碱裂解中的蝶啶鎓盐将芳基引入蝶啶环的简单程序”蝶啶和叶的化学和 liologg 1997 Eds Pfleideier & Rokos，Blackwell Science，Beslin 69-72 (1997)。

T.Sugimoto, C.Seo, and W.Pfleiderer: "A Sinple Procedure to Introduce an Aryl Group into the Pteridine Ring by Arenediazonium Salts in an Alkaline Solution" Chemistry & Biology of Pteridines & Folates, Eds.W.Pfleiderer & H.Rokos, Blackwell Science, Berlin

T.Sugimoto、C.Seo 和 W.Pfleiderer：“在碱性溶液中通过芳重氮盐将芳基引入蝶啶环的简单程序”化学

村田静昭: "Preraration and Synthetic Application of Highly Reactive 6-Trifluoromothanesulfonyloxy-lumazine : A Novel Method for 6-Subatituted Lumazines" Pteridines. 8巻. 1-9 (1997)

Shizuaki Murata：“高反应性 6-三氟甲磺酰氧基二甲氧麻嗪的制备和合成应用：6-取代二甲氧麻嗪的新方法”蝶啶，第 8 卷，1-9 (1997)。

杉本 隆: "Regiosclective Arylation of 1,3-Dimethyllumazine and Ita 5-Oxide by Diazonium Salts" Pteridines. 8巻. 188-194 (1997)

Takashi Sugimoto：“重氮盐对 1,3-二甲基二甲基二氮嗪和 Ita 5-氧化物的区域选择性芳基化”，蝶啶，第 8 卷，188-194 (1997)。