• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

A Screening System for Development of Novel Insulin Secretagogues

Research Project

Project/Area Number 09557075
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 内分泌・代謝学
Research InstitutionChiba University Graduate School of Medicine

Principal Investigator

SEINO Susumu  Chiba University, Graduate School of Medicine, Professor, 大学院・医学研究科, 教授 (80236067)

Co-Investigator(Kenkyū-buntansha) WATANABE Toshifumi  Takeda Chemical Industries, LTD.Pharmaceutical Research Division, Research Fello, 創薬研究本部, 主任研究員
GONOI Tohru  Chiba University, Research Center for Pathogenic Fungi, and Microbial Toxicoses,, 真菌医学研究センター, 助手 (30134365)
MIKI Takashi  Chiba University, Graduate School of Medicine, Assistant, 大学院・医学研究科, 助手 (50302568)
YANO Hideki  Chiba University, Graduate School of Medicine, Associato Professor, 大学院・医学研究科, 教授 (30288576)
渡邉 敏文  武田薬品工業(株), 創薬研究本部, 主任研究員
稲垣 暢也  秋田大学, 医学部, 教授 (30241954)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥11,900,000 (Direct Cost: ¥11,900,000)
Fiscal Year 1998: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1997: ¥8,400,000 (Direct Cost: ¥8,400,000)
KeywordsATP-sensitive K^+ channels / Insulin / sulfonvlurea receptors / inward rectfier / glibenclamide / diabetes mellitus / ATP感受性K^±チャネル / 内向き整流性K^±チャネル
Research Abstract

Development of novel insulin secretagogues is important for the treatment of diabetes mellitus. Sulfonylureas are widely used in the treatment of type 2 diabetes. Sulfonylureas stimulate insulin secretion by inhibiting ATP-sensitive K^+ channels in pancreatic beta-cells. We have recently shown that the beta-cell K_<ATP> channel comprises the inward rectfier K^+ channel subunit Kir6.2 and the sulfonylurea receptor subunit SURI which shows high affinity for the sulfonylurea glibenclamide. We also cloned an isoform of SUR1, called SUR2A which shows low affinity for glibenclamide. To develop screening system of the effectiveness of novel sufonylureas and their derivatives, we have determined 1) the subunit stoichiometry of the beta-cell K_<ATP> channel, 2) sulfonylurea binding sites in the SUR1 ; and 3) we generated Kir6.2 deficient mice.
1). By using fusion proteins of SUR1 and Kir6.2, we found that the activity of K_<ATP> channels is optimized when the the SUR1 subunit and the Kir6.2 subu … More nit are coexpressed with a molar ratio of 1 : 1. Since inward rectifier K^+ channels are thought to function as homo- or hetero-tetramers, this suggests that the beta-cell K_<ATP> channel functions as a hetero- octamer composed of four Kir6.2 subunits and four SUR1 subunits.
2). Various chimeras between SUR1 and SUR2A were prepared. We examined ^3H labeled glibenclamide binding to COS-1 cells transfected with these chimeras and also the effect of glibenclamide on ^<36>Rb efflux from COS-1 cells transfected with each chimera and Kir6.2. We found that high affinity binding site for glibenclamide locates between 15 th and 16th transmembrane segement of SUR1.
3). We generated K_<ATP> channel-deficient mice by genetic disruption of Kir6.2, which forms the K^+ ion-selective pore of the channel. The homozygous mice (Kir6.2^<-/->) lack K_<ATP> channel activity. No significant insulin secretion in response to either glucose or the sulfonylurea tolbutamide was found in K_<ATP> channel-deficient mice (Kir6.2^<-/->), as assessed by perifusion and batch incubation of pancreatic islets. Our studies should provide useful information for the development of new drugs for insulin secretion. Less

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (51 results)

All Other

All Publications (51 results)

  • [Publications] Yanase, H.et al.: "Cellular distribution of sulfonylurea receptor 2 mRNA in the ovary and testic of rats." Biomed.Res.19. 199-204 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sanchez J.A.et al.: "Modulation of reconstituted ATP-sensitive K^+ channels by GTP-binding proteins." J.Physiol.507. 315-324 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Dunne, M.J.et al.: "Familial persistent hyperinsulinemic hypoglycaemia of infancy, and mutations in the sulfonylurea receptor." New Engl.J.Med.336. 703-706 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Inoue, H.et al.: "Sequence variants in the pancreatic islet beta-cell inwardly rectifying K^+ channel Kir6.2(Bir)gene : identification and lack of role in caucasian patients with NIDDM." Diabetes. 46. 502-507 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Suzuki, M.et al.: "Localization of the ATP-sensitive K^+ channel subunit, Kir6.2, in mouse pancreas." Diabetes. 46. 1440-1444 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nestorowicz, A.et al.: "A nonsense mutation in the inward rectifier potassium channel gene, Kir6.2, is associated with familial hyperinsulinism." Diabetes. 46. 1743-1748 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Inagaki, N., Gonoi, T.and Seino, S.: "Subunit stoichiometry of the pancreatic beta-cell ATP-sensitive K^+ channel." FEBS lett.409. 232-236 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ueda, K.et al.: "MgADP antagonism to Mg-independent ATP binding of the sulfonylurea receptor SUR1." J.Biol.Chem.272. 22983-22986 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Miki, T.et al.: "Abnormalities of pancreatic islets by targeted expression of a dominant-negative K_<ATP> channel." Proc.Natl.Acad.Sci.USA. 94. 11969-11973 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Suzuki, M.et al.: "Kir6.1 : a subunit of ATP-sensitive K^+ channels in mitochondria." Biochem.Biophys.Res.Commun.241. 693-637 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sanchez J.A.et al.: "Modulation of reconstituted ATP-sensitive K^+ channels by GTP-binding proteins." J.Physiol.507. 315-324 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yanase, H.et al.: "Cellular distribution of sulfonylurea receptor 2mRNA in the ovary and testis of rats." Biomed.Res.19. 199-204 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Gribble, F.M.et al.: "Tissue specificity of sulfonylureas studies on cloned cardiac and beta-cell K_<ATP> channels." Diabetes. 47. 1412-1418 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Miki, T.et al.: "Defective insulin secretion and enhanced insulin action in K_<ATP> channel-deficient mice." Proc.Natl.Acad.Sci.USA. 95. 10402-10406 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Inagaki, N.& Seino, S.: "ATP sensitive potassium channels : structures, functions, and pathophysiology." Jpn.J.Physiol. 48. 397-412 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ueda, K.et al: "Cooperative binding of ATP and MgADP in the sulfonylurea receptor is modulated by glibenclamide." Proc.Natl.Acad.Sci.USA. 96. 1268-1272 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tanabe, K.et al.: "Direct photoaffinity labeling of the Kir6.2 subunit of the ATP-sensitive K+ channel by 8-azido ATP." J.Biol.Chem.274. 3931-3933 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Miki, T.et al.: "The structure and function of the ATP-sensitive K^+ channel in pancreatic beta-cells." J.Mol.Endo.(in Press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Seino, S.: "ATP-sensitive potassium channels : a model of heteromultimeric potassium channel/receptor assemblies." Annu.Rev.Physiol.(in Press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Gonoi, T.and Seino, S.: "Structure and function of ATP-sensitive K^+ channels.In Pharmacology of Ionic Channel Function : Activators and Inhibitors Springer-Verlng, Heidelberg." Handbook of Experimental Pharmacology. (in Press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Miki, T.et al.: "Structure and function of ATP-sensitive K^+ channels.Academic Press, San Diego, CA" In Potassium Ion Channels : molecular structure, functin, and diseases.(in Press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Seino,S.: "ATP-sensitive potassium channels:a model of heteromultimeric potassium channel/receptor assemblies." Annu.Rev.Physiol.(in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Miki,T.et al.: "The structure and function of the ATP-sensitive K^+ channel in pancreatic β-cells." J.Mol.Endo.(in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ueda,K.et al: "Cooperative binding of ATP and MgADP in the sulfonylurea receptor is modulated by glibenclamide." Proc.Natl.Acad.Sci.USA. 96. 1268-1272 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Tanabe,K.et al.: "Direct photoaffinity labeling of the Kir6.2 subunit of the ATP-sensitive K^+ channel by 8-azido-ATP." J.Biol.Chem.274. 3931-3933 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Inagaki,N.& Seino,S.: "ATP sensitive potassium channels:structures,functions,and pathophysiology." Jpn.J.Physiol.48. 397-412 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Miki,T.et al.: "Defective insulin secretion and enhanced insulin action in K_<ATP> channel-deficient mice." Proc.Natl.Acad.Sci.USA. 95. 10402-10406 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yanase,H.et al.: "Cellular distribution of sulfonylurea receptor 2 mRNA in the ovary and testis of rats." Biomed.Res.19. 199-204 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sanchez J.A.et al.: "Modulation of reconstituted ATP-sensitive K^+ channels by GTP-binding proteins." J.Physiol.507. 315-324 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Suzuki,M.et al.: "Kir6.1:a subunit of ATP-sensitive K^+ channels in mitochondria." Biochem.Biophys.Res.Commun.241. 693-637 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Miki,T.et al.: "Abnormalities of pancreatic islets by targeted expression of a dominant-negative K_<ATP> channel." Proc.Natl.Acad.Sci.USA. 94. 11969-11973 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ueda,K.et al: "MgADP antagonism to Mg-independent ATP binding of the sulfonylurea receptor SUR1." J.Biol.Chem.272. 22983-22986 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Inagaki,N.et al.: "Subunit stoichiometry of the pancreatic β-cell ATP-sensitive K^+ channel." FEBS lett.409. 232-236 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Dunne,M.J.et al.: "Familial persistent hyperinsulinemic hypoglycaemia of infancy,and mutations in the sulfonylurea receptor." New Engl.J.Med.336. 703-706 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Inoue,H.et al.: "Sequence variants in the pancreatic islet β-cell inwardly rectifying K^+ channel Kir6.2(Bir)gene:identification and lack of role in caucasian patients with NIDDM." Diabetes. 46. 502-507 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Suzuki,M.et al.: "Localization of the ATP-sensitive K^+ channel subunit,Kir6.2,in mouse pancreas." Diabetes. 46. 1440-1444 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Nestorowicz,A.et al.: "A nonsense mutation in the inward rectifier potassium channel gene,Kir6.2,is associated with familial hyperinsulinism." Diabetes. 46. 1743-1748 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Gonoi,T.and Seino,S.: "Handbook of Experimental Pharmacology" Springer-Verlng,Heidelberg,(in press), (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Miki,T.et al.: "In Potassium ion channels:molecular structure,functin,and diseases." Academic Press,San Diego,CA,(in press), (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Mizuno,N.et al.: "Altered Bcl-2 and Bax expression and intracellular Ca^<2+> signaling in apoptosis of pancreatic β-cells and the impairment of glucose-induced insulin secretion." Endocrinology(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Sanchez J.A.et al.: "Modulation of reconstituted ATP-sensitive K^+ channels by GTP-binding proteins." J.physiol.(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Suzuki,M.et al.: "Kir6.2:a subunit of ATP-sensitive K^+ channels in mitochondria." Biochem.Biophys.Res.Commun.241. 693-637 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kotake,K.et al.: "Noc2:a putative zinc finger protein involved in excytosis in endocrine cells." J.Biol.Chem.272. 29407-29410 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nestorowicz,A.et al.: "A nonsense mutation in the in ward rectifier potassium channel gene,Kir6.2,is associated with familial hyperinsulinism." Diabetes. 46. 1743-1748 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Mizuta,M.et al.: "Localization and functional role of synaptotagmin III in insulin secretory vesicles in pancreatic β-cells." Diabetes. 46. 2002-2006 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Miki,T.et al.: "Abnormalities of pancreatic islets by targeted expression of a dominant-negative K_<ATP>channel." Proc.Natl.Acad.Sci.USA. 94. 11969-11973 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ueda,K.et al.: "MgADP antagonism to Mg-independent ATP binding of the sulfonylurea receptor SUR1." J.Biol.Chem.272. 22983-22986 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Inagaki,N.et al.: "Subunit stoichiometry of the pancreatic β-cell ATP-sensitive K^+channel." FEBS lett.409. 232-236 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Suzuki,M.et al.: "Localization of the ATP-sensitive K^+channel subunit,Kir6.2,in mouse pancreas." Diabetes. 46. 1440-1444 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Komatsuzaki,K.et al.: "A novel system that reports the G-proteins linked to a given receptor:a study of type 3 somatostatin receptor." FEBS Lett.406. 165-170 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Mizuta,M.et al.: "Suramin inhibits glucose-induced Ca2+ response in single rat pancreatic β cells." Jpn.J.Pharmacol.73. 179-182 (1997)

    • Related Report
      1997 Annual Research Report

URL: 

Published: 1997-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi