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

1998 Fiscal Year Final Research Report Summary

• Project/Area Number: 09640722
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 分離・精製・検出法
• Research Institution: NAGOYA UNIVERSITY
• Principal Investigator: SUGIMOTO Takashi NAGOYA UNIVERSITY,SCHOOL OF INFORMATICS AND SCIENCES,PROFESSOR, 情報文化学部, 教授 (30023609)
• Co-Investigator(Kenkyū-buntansha): MURATA Shizuaki NAGOYA UNIVERSITY,GRADUATE SCHOOL OF HUMAN INFORMATICS,ASSOCIATE PROFESSOR, 大学院・人間情報学研究科, 助教授 (50157781)
• Project Period (FY): 1997 – 1998
• Keywords: enyme linked immunosorbent assay / neopterin / biopterin / arylation / arenediazonium salts / 1.3-dimethyllumazine-6-triflate / nucleophilic substitution with enolates

Research Abstract

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.

Research Products

• [Publications] 杉本 隆: "Arylation of 1,3-Dimethyllumazine by Arenediazonium Salts" Heterocycles.45巻4号. 765-772 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 村田静昭: "Preraration and Synthetic Application of Highly Reactive 6-Trifluoromethanesulfonyloxy-lumazine; A Novel Method for 6-Substituted Lumazines" Pteridines. 8巻. 1-9 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 杉本 隆: "Regioselective Arylation of 1,3-Dimethyllumazine and Its 5-Oxide by Diazonium Salts" Pteridines. 8巻. 188-194 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 村田静昭: "Regioselective Synthesis of 6-Substituted Lumazines by Using Highly Reactive Lumazine 6-Triflate" Chemistry and Biology of Pteridines and Folates 1977,Eds. W.Pfleiderer and H.Rokos,Blackwell Science,Berlin. 17-22 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 杉本 隆: "A Sinple Procedure to Intoruce an Aryl Group into the Pteridine Ring by Arenediazonium Salts in an Alkaline Solution" Chemistry and Biology of Pteridines and Folates 1977,Eds. W.Pfleiderer and H.Rokos,Blackwell Science,Berlin. 69-72 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 村田静昭: "A Novel Method for Introduction of Carbon Substituent into Pteridine" Heterocycles. 50巻1号. 117-124 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] T.Sugimoto and W.Pfleiderer: "Arylation of 1,3-Dimethyllumazine by Arenediazonium Salts" Heterocycles. 45(4). 765-772 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] S.Murata, T.Sugimoto, and K.Murakami: "Preparation and Synthetic Application of Highly Reactive 6-Trifluoromethane sulfonyloxylumazine ; A Novel Method for 6-Substituted Lumazines" Pteridines. 8. 1-9 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] T.Sugimoto, C.Seo, S.Murata, and W.Pfleiderer: "Regioselective Arylation of 1,3-Dimethyllumazine and Its 5-Oxide by Diazonium Salts" Pteridines. 8. 188-194 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 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. 69-72 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] S.Murata, K.Murakami, C.Seo, T.Sugimoto, and M.Kujime: "Regioselective Synthesis of 6-Substituted Lumazines by Using Highly Reactive Lumazine 6-Triflate" Ibid.17-22 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] S.Murata, M.Kujime, T.Sugimoto, K.Murakami, and C.Seo: "A Novel Method for Introduction of Carbon Substituent into Pteridine" Heterocycles. 50(1). 177-124 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] S.Murata, K.Kiguchi, and T.Sugimoto: "Recent Advances in Selective Synthese s of 6-and 7-Substituted Pteridines" Heterocycles. 48(6). 1255-1274 (1998)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] T.Nomura, M.Tazawa, M.Ohtsuki, C.Ichinose, Y.Hagino, A.Ohta, B.Nakashima, K.Mori, T.Sugimoto, Q.Uemo, Y.Nozawa, H.Ichinose, and T.Nagatsu: "Enzymes Related to Catecholamine Biosynthesis in Tetrahymena pyriformis. Presence of GTP Cyclohydrase I" Comparative Biochem.Pysiol.Part B. 120. 753-760 (1998)
  • Description: 「研究成果報告書概要(欧文)」より