1998 Fiscal Year Annual Research Report

細胞周期制御におけるカルシウムシグナリングを含めた新しい情報ネットワークの解明

Research Project

Project/Area Number	09044282
Research Institution	Nagoya University
Principal Investigator	仁木一郎名古屋大学, 医学部, 助教授 (10262908)
Co-Investigator(Kenkyū-buntansha)	LAURENT Meij フランス海洋生物学研究所, 教授内藤康仁名古屋大学, 医学部, 助手 (80303618) 渡邊泰男名古屋大学, 医学部, 講師 (10273228)
Keywords	プロテインキナーゼ阻害剤 / 細胞周期 / イソプリン化合物 / cdc2キナーゼ / イソキノリンスルホナミド化合物
Research Abstract	この2年間の研究交流によって実現した、相互の研究情報の交換および第71回日本生化学会年会のシンポジウムにおけるDr.Meijerの発表「Cyclin-dependent kinases as molecular targets inscrening for new anti-mitoatic agents」などを通して、合成化合物のプロテインキナーゼ阻害剤としての新しい可能性が検討された。この国際学術研究における大きな興味は、これまでHシリーズインヒビターとして用いられてきたイソキノリンスルホナミド化合物が、細胞周期関連キナーゼに対して阻害作用を持つか、という点にあった。その背景には、H-7などのイソキノリンスルホナミド化合物が非一般的なキナーゼ阻害剤として働く可能性が示唆されたからである。今回の共同研究で調べられたイソキノリンスルホナミド系化合物(H-1152およびその誘導体)には、cdc2キナーゼに対してもlowμMオーダーで50%の阻害作用を示した。しかし、Dr.Meijerらが開発したオロモウシンやロスコルビチンなどのイソプロピルプリン誘導体のような選択性に乏しく、両者の阻害様式は、同じATP拮抗型でも異なっていることが示唆された。もうひとつの興味ある問題として、ATF拮抗型キナーゼ阻害剤の阻害作用の強さは、単にそのキナーゼのATP要求性を反映しているのではないか、という点があった。最近報告されたRhoキナーゼに対するイソキノリン化合物(H-7およびHA-1077)などがその例に当てはまる可能性が考えられたからである。しかし、プロテインキナーゼの範囲を拡大して検討した結果、ATP要求性は阻害剤の強さに即反映するわけではないことが判明した。これらの研究成果は、新たなキナーゼ阻害剤の開発と、その薬剤への応用を考えるうえでも重要なポイントである。

Research Products

(27 results)

All Other

All Publications (27 results)

[Publications] Iida Y.: "Myosin light chain phosphorylation controls insulin secretion at a proximal step in the secretory cascade." Am.J.Physiol.(Endocrinol.Metab.36). 273. E789-E782 (1997)
[Publications] Kabir N.: "Immunocytochemical detection and spatial distribution of myosin light-chain kinase in preimplantation mouse embryos." J.Exp.Zool.278. 147-155 (1997)
[Publications] Watanabe Y.: "Identification of a specific amino acid cluster in the calmodulin-binding domain of the neuronal nitric oxide synthase" FEBS Lett.403. 75-78 (1997)
[Publications] Naito Y.: "Isoform-specific activation and structural diversity of CaM kinase I" J.Biol.Chem.272. 32704-32708 (1997)
[Publications] Yokokura H.: "Isolation and comparison of rat cDNAs encoding Ca^<2+>/calmodulin-dependent protein kinase I isoforms." Biochim.Biophys.Acta(Short sequence paper). 1338. 8-12 (1997)
[Publications] Hidaka H.: "Cascade activation of the calmodulin kinase family." in Signal Transduction in Health and Disease,edited by J.Corbin Raven Press.New York,U.S.A.(Proceedings of the 9th International Conference on Second Messengers and Phosphoproteins.Oct.27-Nov.1.1995.Nashville.U.S.A.). 151-157 (1997)
[Publications] 渡邊泰男: "NOSの分子生物学的解析による構造機能活性相関ならびに活性化メカニズムの検討" 日本薬理学雑誌(Folia Pharmacol.Jpn.). 110. 69-75 (1997)
[Publications] Schutte B.: "The effect of the cyclin-dependent kinase inhibitor olomoucine on cell cycle kinetics." Exp Cell Res.236・1. 4-15 (1997)
[Publications] De Azevedo WF.: "Inhibition of cyclin-dependent kinases by purine analogues: crystal structure of human cdk2 complexed with roscovitine." Eur.J.Biochem.243・1-2. 518-526 (1997)
[Publications] Meijer L.: "Biochemical and cellular effects of roscovitine,a potent and selective inhibitor of the cyclin-dependent kinases cdc2,cdk2 and cdk5." Eur.J.Biochem.243・1-2. 527-536 (1997)
[Publications] Havlicek L.: "Cytokinin-derived cyclin-dependent kinase inhibitors: synthesis and cdc2 inhibitory activity of olomoucine and related compounds." J.Med.Chem.40・4. 408-412 (1997)
[Publications] Pippin JW.: "Direct in vivo inhibition of the nuclear cell cycle cascade in experimental mesangial proliferative glomerulonephritis with Roscovitine,a novel cyclin-dependent kinase antagonist." J.Clin.Invest.100・10. 2512-2520 (1997)
[Publications] Krucher NA.: "The cyclin-dependent kinase (cdk) inhibitors,olomoucine and roscovitine,alter the expression of a molluscan circadian pacemaker." Cell Mol.Neurobiol.17・5. 495-507 (1997)
[Publications] Meijer L.: "Chemical inhibitors of cyclin-dependent kinases." Methods Enzymol.283. 113-128 (1997)
[Publications] Planchais S.: "Roscovitine,a novel cyclin-dependent kinase inhibitor,characterizes restriction point and G2/M transition in tobacco BY-2 cell suspension." Plant J.12・1. 191-202 (1997)
[Publications] Watanabe H.: "An essential role of myosin light-chain kinase in the regulation of agonist-and fluid-flow-stimulated Ca^<2+> influx in endothelial cells." FASEBJ.12. 341-348 (1998)
[Publications] Niwa T.^-: "Acetylcholine activates intracellular movement of insulin granules in the pancreatic β-cells via IP3-dependent mobilization of intracellular Ca^<2+>" Diabetes. 47. 1699-1706 (1998)
[Publications] 仁木一郎: "インスリン顆粒運動の制御と細胞内Ca^<2+>の分業仮説." Peptide Hormones in Pancreas. 18. 112-116 (1998)
[Publications] Watanabe Y.: "Neuronal nitric oxide synthase-membrane phospholipid interaction" Arch.Biochem.Biophys.358. 68-73 (1998)
[Publications] Watanabe Y.: "Inter-isoformal regulation of nitric oxide synthase through heteromeric dimerization" Biochim.Biophys.Acta. 1388. 199-208 (1998)
[Publications] Nishio M.: "HMN-1180,a small molecule inhibitor of neuronal nitric oxide synthase" J.Pharmacol.Exp.Ther.287. 1063-1067 (1998)
[Publications] Sakaguchi H.: "Unique inhibitory action of the synthetic compound 2-[N-(2-aminoethyl)-N-(5-isoquinolinesulfonyl)]amino-N-(4-chlorocinnamyl)-N-methylbenzylamine(CKA-1306)against calcium/calmodulin-dependent protein kinase I" Biochem.Pharmacol.56. 329-334 (1998)
[Publications] Gray NS.: "Exploiting chemical libraries,structure and genomics in the search for kinase inhibitors." Science. 281・5376. 533-538 (1998)
[Publications] Niki I.: "Intracellular Ca^<2+> receptors in the pancreatic β-cell." Molec.Cell Biochem.(in print). (1998)
[Publications] Niki I.: "Ca^<2+> signalling and the insulin secretory cascade in pancreaticbeta-cell." Jap.J.Pharmacol.(in print). (1999)
[Publications] Ono-Saito N.: "H-series protein kinase inhibitors and a case of a H-series inhibitor for clinical application." Pharmacol.Therap.(in print). (1999)
[Publications] 丹羽多恵: "グルコースによるインスリン分泌顆粒運動の活性化機構." Peptide Hormones in Pancreas. (in print). (1999)

1998 Fiscal Year Annual Research Report

細胞周期制御におけるカルシウムシグナリングを含めた新しい情報ネットワークの解明

Principal Investigator

仁木 一郎 名古屋大学, 医学部, 助教授 (10262908)

Research Products

[Publications] Iida Y.: "Myosin light chain phosphorylation controls insulin secretion at a proximal step in the secretory cascade." Am.J.Physiol.(Endocrinol.Metab.36). 273. E789-E782 (1997)

[Publications] Kabir N.: "Immunocytochemical detection and spatial distribution of myosin light-chain kinase in preimplantation mouse embryos." J.Exp.Zool.278. 147-155 (1997)

[Publications] Watanabe Y.: "Identification of a specific amino acid cluster in the calmodulin-binding domain of the neuronal nitric oxide synthase" FEBS Lett.403. 75-78 (1997)

[Publications] Naito Y.: "Isoform-specific activation and structural diversity of CaM kinase I" J.Biol.Chem.272. 32704-32708 (1997)

[Publications] Yokokura H.: "Isolation and comparison of rat cDNAs encoding Ca^<2+>/calmodulin-dependent protein kinase I isoforms." Biochim.Biophys.Acta(Short sequence paper). 1338. 8-12 (1997)

[Publications] 渡邊泰男: "NOSの分子生物学的解析による構造機能活性相関ならびに活性化メカニズムの検討" 日本薬理学雑誌(Folia Pharmacol.Jpn.). 110. 69-75 (1997)

[Publications] Schutte B.: "The effect of the cyclin-dependent kinase inhibitor olomoucine on cell cycle kinetics." Exp Cell Res.236・1. 4-15 (1997)

[Publications] De Azevedo WF.: "Inhibition of cyclin-dependent kinases by purine analogues: crystal structure of human cdk2 complexed with roscovitine." Eur.J.Biochem.243・1-2. 518-526 (1997)

[Publications] Meijer L.: "Biochemical and cellular effects of roscovitine,a potent and selective inhibitor of the cyclin-dependent kinases cdc2,cdk2 and cdk5." Eur.J.Biochem.243・1-2. 527-536 (1997)

[Publications] Havlicek L.: "Cytokinin-derived cyclin-dependent kinase inhibitors: synthesis and cdc2 inhibitory activity of olomoucine and related compounds." J.Med.Chem.40・4. 408-412 (1997)

[Publications] Pippin JW.: "Direct in vivo inhibition of the nuclear cell cycle cascade in experimental mesangial proliferative glomerulonephritis with Roscovitine,a novel cyclin-dependent kinase antagonist." J.Clin.Invest.100・10. 2512-2520 (1997)

[Publications] Krucher NA.: "The cyclin-dependent kinase (cdk) inhibitors,olomoucine and roscovitine,alter the expression of a molluscan circadian pacemaker." Cell Mol.Neurobiol.17・5. 495-507 (1997)

[Publications] Meijer L.: "Chemical inhibitors of cyclin-dependent kinases." Methods Enzymol.283. 113-128 (1997)

[Publications] Planchais S.: "Roscovitine,a novel cyclin-dependent kinase inhibitor,characterizes restriction point and G2/M transition in tobacco BY-2 cell suspension." Plant J.12・1. 191-202 (1997)

[Publications] Watanabe H.: "An essential role of myosin light-chain kinase in the regulation of agonist-and fluid-flow-stimulated Ca^<2+> influx in endothelial cells." FASEBJ.12. 341-348 (1998)

[Publications] Niwa T.^-: "Acetylcholine activates intracellular movement of insulin granules in the pancreatic β-cells via IP3-dependent mobilization of intracellular Ca^<2+>" Diabetes. 47. 1699-1706 (1998)

[Publications] 仁木一郎: "インスリン顆粒運動の制御と細胞内Ca^<2+>の分業仮説." Peptide Hormones in Pancreas. 18. 112-116 (1998)

[Publications] Watanabe Y.: "Neuronal nitric oxide synthase-membrane phospholipid interaction" Arch.Biochem.Biophys.358. 68-73 (1998)

[Publications] Watanabe Y.: "Inter-isoformal regulation of nitric oxide synthase through heteromeric dimerization" Biochim.Biophys.Acta. 1388. 199-208 (1998)

[Publications] Nishio M.: "HMN-1180,a small molecule inhibitor of neuronal nitric oxide synthase" J.Pharmacol.Exp.Ther.287. 1063-1067 (1998)

[Publications] Sakaguchi H.: "Unique inhibitory action of the synthetic compound 2-[N-(2-aminoethyl)-N-(5-isoquinolinesulfonyl)]amino-N-(4-chlorocinnamyl)-N-methylbenzylamine(CKA-1306)against calcium/calmodulin-dependent protein kinase I" Biochem.Pharmacol.56. 329-334 (1998)

[Publications] Gray NS.: "Exploiting chemical libraries,structure and genomics in the search for kinase inhibitors." Science. 281・5376. 533-538 (1998)

[Publications] Niki I.: "Intracellular Ca^<2+> receptors in the pancreatic β-cell." Molec.Cell Biochem.(in print). (1998)

[Publications] Niki I.: "Ca^<2+> signalling and the insulin secretory cascade in pancreaticbeta-cell." Jap.J.Pharmacol.(in print). (1999)

[Publications] Ono-Saito N.: "H-series protein kinase inhibitors and a case of a H-series inhibitor for clinical application." Pharmacol.Therap.(in print). (1999)

[Publications] 丹羽多恵: "グルコースによるインスリン分泌顆粒運動の活性化機構." Peptide Hormones in Pancreas. (in print). (1999)

仁木一郎名古屋大学, 医学部, 助教授 (10262908)