生物新機能と創薬をめざす生体内分子科学

2002 Fiscal Year Annual Research Report

• Project/Area Number: 13NP0401
• Research Institution: Gifu University
• Principal Investigator: 鈴木 正昭 岐阜大学, 大学院・医学研究科, 教授 (90093046)
• Co-Investigator(Kenkyū-buntansha): 石川 智久 東京工業大学, 大学院・生命理工学研究科, 教授 (60193281) ; 松田 彰 北海道大学, 大学院・薬学研究科, 教授 (90157313) ; 渡辺 恭良 大阪市立大学, 大学院・医学研究科, 教授 (40144399) ; 米倉 義晴 福井医科大学, 高エネルギー医学研究センター, 教授 (60135572) ; 森 望 国立長寿医療研究センター, 分子遺伝学研究部, 部長(研究職) (00130394)
• Keywords: プロスグランジン / RXRアンタゴニスト / 可視化プローブ / ABCトランスポーター / 4'-チオリボヌクレオシド / ダントロレン誘導体 / Cu-ATSM / PET

Research Abstract

今年度に達成されたの主な具体的成果を箇条書きにした。
(1)分子プローブ設計グループ
神経突起伸展促進および細胞死抑制作用を示す新規エノン型プロスタグランジン(PG)の創製に成功した。これらの作用にはPGによるHO-1の誘導やJNKの活性化抑制が関与していることを明らかにした。核内受容体RXRヘテロダイマーのアンタゴニストHX531およびPA452を創製した。HX531はPPARγ-RXRの機能を阻害し、マウスにおいて抗糖尿病、抗肥満効果を示した。糖部をN型およびS型に強く束縛した新規核酸アナログを合成し、これらを組み込んだ修飾型オリゴヌクレオチドの創製に成功した。4'-チオリボヌクレオシドの立体選択的大量合成法を確立し、これらを組み込んだ4'-チオRNAオリゴマーの合成に成功した。生細胞可視化プローブに関して、フルオレスセイン類の構造と蛍光特性との関係を明らかにした。グルタミン酸トランスポーターブロッカーL-TBOAの構造を基に、グルタミン酸受容体作用が無く、より親和性の高い誘導体TFB-TBOAおよび光感受性のcaged-TBOAの創製に成功した。カップリング反応によるダントロレン誘導体の一般的合成法を確立し、生理的Ca^<2+>放出やCa^<2+>によるCa^<2+>放出機構に特異的に作用する探索分子の創製に成功した。
(2)分子機能解析グループ
ABCトランスポーター基質特異性解析のハイスループット化をめざし、ヒトP-糖タンパク質(ABCBl)の昆虫細胞での発現方法の確立と、96wellプレートを用いたATPase活性測定高速スクリーニング系の構築に成功した。これを用いて医薬品約50種をスクリーニングし、構造-輸送活性相関データベースを構築した。
(3)生体機能評価グループ
Shc、N-Shc、Sckのリン酸化チロシン受容ドメイン、SH2ドメインでのペプチドリガンドとの結合様式を分子モデリング手法を使って解析した。ベンゾチアゾール系化合物TG003がSRタンパク質リン酸化酵素のClk/StylおよびClk/Sty4を阻害することで選択的スプライシングのパターンに影響を与えることを見いだした。プロスタグランジンE受容体サブタイプEP3に共役するGタンパク質G12ファミリーに結合するエフェクター分子としてセリン・スレオニンホスファターゼPP5のクローニングに成功した。
(4)分子イメージンググループ
低酸素腫瘍に特異的に集積するPETトレーサーCu-ATSMの創製に成功した。[^<18>F]FDGと[^<64>Cu]ATSMを用いた担癌ウサギにおけるPET画像の比較から、両者に特徴的な集積が観測され、腫瘍画像診断における相補的検査手法となる可能性を示した。バイオラジオグラフィー法において時間解像度を改善するためPositron Emission Photon Imaging(PEPI)法を開発し、2分ごとの脳スライスイメージングを可能にした。

Research Products

• [Publications] M.Ikenoya: "Inhibition of Rho-kinase-induced myristoylated alanine-rich C kinase substrate (MARCKS) phosphorylation in human neuronal cells by H-1152, a novel and specific Rho-kinase inhibitor"J.Neurochem.. 81・1. 9-16 (2002)
• [Publications] Y.Sasaki: "The novel and specific Rho-kinase inhibitor (S)-(+)-2-methyl-1-{(4-methyl-5-insoquinoline)sulfonyl}-homopiperazine as a probing molecule for Rho-kinase-involved pathway"Pharmacology & Therapeutics. 93・. 225-232 (2002)
• [Publications] T.Hosoya: "[^<125>I]-N-[(3-Azido-5-iodo)benzyl]dantrolene and [^<125>I]-N-{[3-iodo-5-(3-trifuoromethyl-3H-diazirin-3-yl)]benzyl}dantrolene : Photoaffinity Probes Specific for the Physiological Ca^<2+> Release from Sarcoplasmic Reticulum of Skeletal Muscle"Bioorg.Med.Chem.Lett.. 12・22. 3263-3265 (2002)
• [Publications] T.Hosoya: "Dantrolene Analogues Revisited : General Synthesis and Specific Functions Capable of Discriminating Two Kinds of Ca^<2+> Release from Sarcoplasmic Reticulum of Mouse Skeletal Muscle"Bioorg.Med.Chem.Lett.. 11・5. 663-673 (2003)
• [Publications] 鈴木正昭: "学術創成研究「生物新機能と創薬をめざす生体内分子科学」-化学・生物学・医学融合型研究プロジェクト-"MEDCHEM NEWS (essay). 12・9. 9-12 (2002)
• [Publications] 萩原正敏: "序 新たなパラダイムを拓く分子プローブの創製:生物学と化学の融合 (特集 分子プローブで変わる生物学-生命機能の解明から創薬まで)"細胞工学. 22・2. 134-135 (2003)
• [Publications] 鈴木正昭: "設計PGプローブによるヒト脳内IP_2受容体の分子イメージング (特集 分子プローブで変わる生物学-生命機能の解明から創薬まで)"細胞工学. 22・2. 165-171 (2003)
• [Publications] T.Kodama: "A highly stereoselective samarium diiodide-promoted aldol reaction with 1'-phenylseleno-2'-keto nucleosides. Synthesis of l'α-branched uridine derivatives"J.Org.Chem.. 67・22. 7706-7715 (2002)
• [Publications] Y.Komatsu: "In vitro selection of hairpin ribozymes activated with short oligonucleotides"Biochemistry. 41・29. 9090-9098 (2002)
• [Publications] H.A.Guse: "Novel hydrolysis-resistant analogues of cyclic ADP-ribose : modification of the "northern" ribose and calcium release activity"Biochemistry. 41・21. 6744-6751 (2002)
• [Publications] Y.Shimamoto: "Cellular and Biochemical Mechanisms of the Resistance of Human Cancer Cells to a New Anticancer Ribo-nucleoside, TAS-106"Jpn.J.Cancer Res.. 93・4. 445-452 (2002)
• [Publications] Y.Kazuta: "Synthesis of (1S,2R)-1-phenyl-2-[(S)-l-aminopropyl]-N, N-diethylcyclopropanecarboxamide (PPDC) derivatives modified at the carbamoyl moiety as a new class of NMDA receptor antagonists"Bioorg.Med.Chem.. 10・6. 1777-1791 (2002)
• [Publications] Y.Kazuta: "Development of versatile cis-and trans-dicarbon -substituted chiral cyclopropane units : synthesis of (1S,2R)-and (1R,2R)-2-aminomethyl-1-(1H-imidazol-4-yl)cyclopropanes and their enantiomers as conformationally restricted analogues of histamine"J.Org.Chem.. 67・5. 1669-1677 (2002)
• [Publications] S.Shuto: "New neplanocin analogues. 12. Alternatives synthesis and antimalarial effect of (6'R)-6'-C-methylneplanocin A, a potent AdoHcy hydrolase inhibitor"J.Med.Chem.. 45・3. 748-751 (2002)
• [Publications] M.Nomura: "Nucleosides and nucleotides. Part 212: Practical large-scale synthesis of 1-(3-C-ethynyl-β-D-ribo-pentofuranosyl)cytosine (ECyd), a potent antitumor nucleoside. Isobutyryloxy group as an efficient anomeric leaving group in the Vorbruggen glycosylation reaction"Tetrahedron. 58・7. 1279-1288 (2002)
• [Publications] H.Sumiyoshi: "Solution-Phase Parallel Synthesis of Carbamates Using Polymer-Bound N-Hydroxysuccinimide"Org.Lett.. 4・22. 3923-3926 (2002)
• [Publications] K.Hanaoka: "Design and synthesis of a novel magnetic resonance imaging contrast agent for selective sensing of zinc ion"Chem.Biol.. 9・9. 1027-1032 (2002)
• [Publications] S.Maruyama: "A novel, cell-permeable, fluorescent probe for ratiometric imaging of zinc ion"J.Am.Chem.Soc.. 124・36. 10650-10651 (2002)
• [Publications] N.Suzuki: "Multiple active intermediate in oxidation reaction catalyzed by synthetic hemethiolate complex relevant to cytochrome p450"J.Am.Chem.Soc.. 124・32. 9622-9628 (2002)
• [Publications] S.Ueno: "Mossy fiber Zn^<2+> spillover modulates heterosynaptic N-methyl-D-aspartate receptor activity in hippocampal CA3 circuits"J.Cell Biol.. 158・2. 215-220 (2002)
• [Publications] T.Hirano: "Improvement and biological applications of fluorescent probes for zinc, ZnAFs"J.Am.Chem.Soc.. 124・23. 6555-6562 (2002)
• [Publications] S.Mizukami: "A fulorescent anion sensor that works in neutral aqueous solution for bioanalytical application"J.Am.Chem.Soc.. 124・15. 3920-3925 (2002)
• [Publications] T.Nagano: "Bioimaging of nitric oxide"Chem.Rev.. 102・4. 1235-1270 (2002)
• [Publications] H.Takakusa: "Design and synthesis of an enzyme-cleavable sensor molecule for phosphodiesterase activity based on fluorescence resonance energy transfer"J.Am.Chem.Soc.. 124・8. 1653-1657 (2002)
• [Publications] M.Nishida: "Activation mechanism of Gi and Go by reactive oxygen species"J.Biol.Chem.. 277・11. 9036-9042 (2002)
• [Publications] S-J.Ma: "A direct continuous spectrophotometric assay for glycosidases with 3-nitro-2-pyridylglycosides by tautomerization of 2-hydroxy-3-nitropyridine"Anal.Biochem.. 302・2. 291-297 (2002)
• [Publications] H.Akamatsu: "New efficient route for solid-phase synthesis of bensimidazole derivatives"J.Comb.Chem.. 4・5. 475-783 (2002)
• [Publications] M.Wakao: "Chemical synthesis of cyclodextrins by using intramolecular glycosylation"J.Org.Chem.. 67・23. 8182-8190 (2002)
• [Publications] M.Izumi: "Solid-phase synthesis of oligosaccharides using novel alkyne-type linkers : selection of reactive sites on the support by Sonogashira reaction"Synlett.. 9. 1409-1416 (2002)
• [Publications] H.Akamatsu: "Reactivity switching on solid support : solid-phase synthesis of teritary amines by reduction of tertiary amides with LiAlH_4"Tetrahedron Lett.. 43・49. 8867-8869 (2002)
• [Publications] Y.Hashimoto: "Structural development of biological response modifiers based on thalidomide"Bioorg.Med.Chem.. 10・3. 461-479 (2002)
• [Publications] T.Ishioka: "Novel non-steroidal/non-anilide type androgen antagonists with an isoxazolone moiety"Bioorg.Med.Chem.. 10・5. 1555-1566 (2002)
• [Publications] T.Noguchi: "Thalidomide and its analog as cyclooxygenase inhibitors"Bioorg.Med.Chem.Lett.. 12・7. 1043-1046 (2002)
• [Publications] 加藤優子: "新規窒素含有ビタミンD_3誘導体の合成"ビタミン. 76・3. 151-151 (2002)
• [Publications] 橋本祐一: "サリドマイドのマルチ創薬テンプレートとしての活用一ドラマタイプ創薬の1例"Bioベンチャー. 2・5. 43-49 (2002)
• [Publications] Y.Hashimoto: "Structural development of biological response modifiers based on retinoids and thalidomide"Mini-Rev.Med.Chem.. 2・6. 543-552 (2002)
• [Publications] H.Kakuta: "Fluorescent bioprobes for visualization of puromycin-sensitive aminopeptidase in living cells"Bioorg.Med.Chem.Lett.. 13・1. 83-86 (2002)
• [Publications] Y.Hashimoto: "Structural development of synthetic retinoids and thalidomide-related molecules"Cancer Chemother. Pharmacol.. (印刷中).
• [Publications] K.Nagasawa: "Total synthesisn of crambescidin 359"Org.Lett.. 4・2. 177-180 (2002)
• [Publications] H.Takahashi: "Novel Mannich-type nucleophilic substitution reaction with tertiary aromatic amines"Tetrahedron Lett.. 43・16. 2935-2938 (2002)
• [Publications] N.Morisaki: "Coordination of sodium cation to an oxygen function and olefinic double bond to form molecular adduct ion in fast bombardment mass spectroscopy"Chem.Pharm.Bull.. 50・7. 935-940 (2002)
• [Publications] H.Takahashi: "Nucleophilic substitution on 4-hydroxymethylanilines under neutral conditions via aza quinone methide intermediate"Tetrahedron Lett.. 43・33. 5751-5753 (2002)
• [Publications] B.Takahashi: "Novel retinoid X receptor antagonists : Specific inhibitors of retinoid synergism in RXR-RAR heterodimer actions"J.Med.Chem.. 45・16. 3227-3330 (2002)
• [Publications] T.Kita: "C_2-Symmetric chiral pentacyclic guanidine : A phase-transfer catalyst for the asymmetric alkylation of tert-butyl glycinate Schiff base"Angew.Chem.Int.Ed.. 41・15. 2832-2834 (2002)
• [Publications] T.Ishiwata: "Total synthesis of batazelladine D"Org.Lett.. 4・17. 2921-2924 (2002)
• [Publications] K.Nagasawa: "Stereoselective synthesis of trycyclic guanidine, the key component of the batzelladin alkaloids"Tetrahedron Lett.. 43・36. 6383-6385 (2002)
• [Publications] H.Kagechika: "Novel Synthetic Retinoids and Separation of the Pleiotropic Retinoidal Activities"Curr.Med.Chem.. 9・5. 591-608 (2002)
• [Publications] T.Imanishi: "BNAs : novel nucleic acid analogs with a bridged sugar moiety"Chem.Commun.(Camb).. 16. 1653-1659 (2002)
• [Publications] K.Morita: "2'-O, 4'-C-ethylene-bridged nucleic acids (ENA) : highly nuclease-resistant and thermodynamically stable oligonucleotides for antisense drug"Bioorg.Med.Chem.Lett.. 12・1. 73-76 (2002)
• [Publications] T.Imanishi: "Down-regulation of a gene-expression by an antisense BNA oligonucleotide"Nippon Yakurigaku Zasshi. 120・2. 85-90 (2002)
• [Publications] S.Obika: "Synthesis and conformation of a novel bridged nucleoside with S-type sugar puckering, trans-3',4'-BNA monomer"Tetrahedron Lett.. 43・24. 4365-4368 (2002)
• [Publications] S.Obika: "Synthesis and conformation of 3',4'-BNA monomers, 3'-O, 4'-C methyleneribonucleosides"Tetrahedron. 58・15. 3093-3049 (2002)
• [Publications] Y.Hari: "Effecttive synthesis of C-nucleosides with 2',4'-BNA modification"Tetrahedron. 58・15. 3051-3063 (2002)
• [Publications] M.Yoshikawa: "Molecular and cytogenetic characterization of the mouse ATP-binding cassette transporter Abcg4"Gene. 293・1-2. 67-75 (2002)
• [Publications] H.Yabuuchi: "ABCC13, an unusual truncated ABC transporter, is highly expressed in fetal human liver"Biochem.Biophys.Res.Commun.. 299・3. 410-417 (2002)
• [Publications] H.Shimizu: "Characterization of the mouse Abcc12 gene and its transcript encoding an ATP-binding cassette transporter, an orthologue of human ABCC12"Gene. (印刷中).
• [Publications] Y.Hosaka: "α1-synthrophin-deficient skeletal muscle exhibits hypertrophy and aberrant formation of neuromuscular junctions during regeneration"J.Cell.Biol.. 158・6. 1097-1107 (2002)
• [Publications] M.Sakamoto: "Micro-dystrophin cDNA ameliorates dystrophic phenotypes when introduced into mdx mice as a transgene"Biochem.Biophys.Res.Commun.. 293・4. 1265-1272 (2002)
• [Publications] Y.Yamaguchi: "G12 and G13 interact with Ser/Thr protein phosphatase type 5 and stimulate its phosphatase activity"Curr.Biol.. 12・15. 1353-1358 (2002)
• [Publications] M.Negishi: "Rho family GTPases as key regulators for neuronal network formation"J.Biochem.(Tokyo). 132・2. 157-166 (2002)
• [Publications] K.Nakamura: "The rostral raphe pallidus nucleus mediates pyrogenic transmission from the preoptic area"J.Neurosci.. 22・11. 4600-4610 (2002)
• [Publications] H.Okada: "Distinct roles of activating transcription factor 6 (ATF6) and double-stranded RNA-activated protein kinase-like endoplasmic reticulum kinase (PERK) in transcription during the mammalian unfolded protein response"Biochem.J.. 366・Pt2. 585-594 (2002)
• [Publications] A.Mouihate: "Suppression of fever at near term is associated with reduced COX-2 protein expression in rat hypothalamus"Am.J.Physiol.Regul.Integr.Comp.Physiol.. 283・3. R800-805 (2002)
• [Publications] H.Katoh: "Socius is a novel Rnd GTPase-interacting protein involved in disassembly of actin stress fibers"Mol.Cell.Biol.. 22・9. 2952-2964 (2002)
• [Publications] H.Tanaka: "Vps4-A (vacuolar protein sorting 4-A) is a binding partner for a novel Rho family GTPase, Rnd2"Biochem.J.. 365・Pt2. 349-353 (2002)
• [Publications] N.Hatae: "Augmentation of receptor-mediated adenylyl cyclase activity by Gi-coupled prostaglandin receptor subtype EP3 in a Gbetagamma subunit-independent manner"Biochem.Biophys.Res.Commun.. 290・1. 162-168 (2002)
• [Publications] Y.Hippo: "Global gene expression analysis of gastric cancer by oligonucleotide microarrays"Cancer Research. 62・1. 233-240 (2002)
• [Publications] A.Mukasa: "Distinction in gene expression profiles of oligodendrogliomas with and without alleic loss of 1p"Oncogene. 21・25. 3961-3968 (2002)
• [Publications] T.Ota: "Targets of transcriptional regulation by two distinct type I receptors for transforming growth factor-beta in human umbilical vein endothelial cells"J.Cell.Physiol.. 193・3. 299-318 (2002)
• [Publications] Y.Fujiwara: "Influence of polyunsaturated fatty acid on gene expression in HepG2 cells"J.Anteroscler.Thromb.. 9・3. 157-162 (2002)
• [Publications] J.M.Chong: "Interleukin-1 beta Expression in Human Gastric Carcinoma with Epstein-Barr Virus Infection"J.Virol.. 76・13. 6825-6831 (2002)
• [Publications] A.Saiura: "Detection of an up-regulation of a group of chemokine genes in murine cardiac allograft in the absence of interferon-gammma by means of DNA microarray 1"Transplantation. 73・9. 1480-1486 (2002)
• [Publications] Y.Midorikawa: "Identification of genes associated with de differentiation of hepatocellular carcinoma with expression profiling analysis"Jpn.J.Cancer Res.. 93・6. 636-643 (2002)
• [Publications] M.Takahashi: "Fish oil feeding alters liver gene expressions to defend against PPARalpha activation and ROS production"Am.J.Physiol.Gastrointest Liver Physiol.. 282・2. G338-348 (2002)
• [Publications] T.Nakamura: "Grit, a GTPase-activating protein for the Rho family, regulates neurite extension through association with the TrkA receptor and N-Shc and CrkL/Crk adapter molecules"Mol.Cell Biol.. 22・24. 8721-8734 (2002)
• [Publications] T.Nakazawa: "Comparative expression profiles of Trk receptors and Shc-related phosphotyrosine adapters during retinal development : potential roles of N-Shc/ShcC in brain-derived neurotrophic factor signal transduction and modulation"J.Neurosci.Res.. 68・6. 668-680 (2002)
• [Publications] T.Nakamura: "Discrimination between phosphotyrosine-mediated signaling properties of conventional and neuronal Shc adapter molecules"Oncogene. 21・1. 22-31 (2002)
• [Publications] T.Muratani: "Characterization of nociceptin/orphanin FQ-induced pain responses by the novel receptor antagonist N-(4-amino-2-methylquinolin-6-yl)-2-(4-ethylphenoxymethyl)benzamide monohydrochloride"J.Pharamocol.Exp.ther.. 303・1. 424-430 (2002)
• [Publications] M-R.Zhang: "Development of an automated system for synthesizing ^<18>F-labeled compounds using [^<18>F]fluoroethyl bromide as a synthetic precursor"Appl.Radiat.Isot.. 57・3. 335-342 (2002)
• [Publications] M-R.Zhang: "Synthesis and preliminary evaluation of [^<18>F]FEtP4A, a promising PET tracer for mapping acetylcholinesterase in vivo"Nucl.Med.Biol.. 29・4. 463-468 (2002)
• [Publications] H.Tsukada: "Comparative effects of methamphetamine and nicotine on the striatal [^<11>C]raclopride binding in unanesthetized monkeys"Synapse. 45・4. 207-212 (2002)
• [Publications] N.Harada: "Age-related changes in the striatal dopaminergic system in the living brain : a multiparametric PET study in conscious monkeys"Synapse. 45・1. 38-45 (2002)
• [Publications] N.Harada: "Age differences in phosphodiesterase type-IV and its functional response to dopamine D_1 receptor modulation in the living brain : a PET study in conscious monkeys"Synapse. 44・3. 139-145 (2002)
• [Publications] K.Sakurada: "Synapsin I is phosphorylated at Ser603 by p21-activated kinases (PAKs) in vitro and in PC21 cells stimulated with bradykinin"J.Biol.Chem.. 277・47. 45473-45479 (2002)