生物新機能と創薬をめざす生体内分子科学

2002 年度 実績報告書

• 研究課題/領域番号: 13NP0401
• 研究機関: 岐阜大学
• 研究代表者: 鈴木 正昭 岐阜大学, 大学院・医学研究科, 教授 (90093046)
• 研究分担者: 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)
• キーワード: 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

研究概要

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。

研究成果

• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] Y.Komatsu: "In vitro selection of hairpin ribozymes activated with short oligonucleotides"Biochemistry. 41・29. 9090-9098 (2002)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] H.Sumiyoshi: "Solution-Phase Parallel Synthesis of Carbamates Using Polymer-Bound N-Hydroxysuccinimide"Org.Lett.. 4・22. 3923-3926 (2002)
• [文献書誌] 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)
• [文献書誌] S.Maruyama: "A novel, cell-permeable, fluorescent probe for ratiometric imaging of zinc ion"J.Am.Chem.Soc.. 124・36. 10650-10651 (2002)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] T.Hirano: "Improvement and biological applications of fluorescent probes for zinc, ZnAFs"J.Am.Chem.Soc.. 124・23. 6555-6562 (2002)
• [文献書誌] S.Mizukami: "A fulorescent anion sensor that works in neutral aqueous solution for bioanalytical application"J.Am.Chem.Soc.. 124・15. 3920-3925 (2002)
• [文献書誌] T.Nagano: "Bioimaging of nitric oxide"Chem.Rev.. 102・4. 1235-1270 (2002)
• [文献書誌] 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)
• [文献書誌] M.Nishida: "Activation mechanism of Gi and Go by reactive oxygen species"J.Biol.Chem.. 277・11. 9036-9042 (2002)
• [文献書誌] 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)
• [文献書誌] H.Akamatsu: "New efficient route for solid-phase synthesis of bensimidazole derivatives"J.Comb.Chem.. 4・5. 475-783 (2002)
• [文献書誌] M.Wakao: "Chemical synthesis of cyclodextrins by using intramolecular glycosylation"J.Org.Chem.. 67・23. 8182-8190 (2002)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] Y.Hashimoto: "Structural development of biological response modifiers based on thalidomide"Bioorg.Med.Chem.. 10・3. 461-479 (2002)
• [文献書誌] T.Ishioka: "Novel non-steroidal/non-anilide type androgen antagonists with an isoxazolone moiety"Bioorg.Med.Chem.. 10・5. 1555-1566 (2002)
• [文献書誌] T.Noguchi: "Thalidomide and its analog as cyclooxygenase inhibitors"Bioorg.Med.Chem.Lett.. 12・7. 1043-1046 (2002)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] Y.Hashimoto: "Structural development of biological response modifiers based on retinoids and thalidomide"Mini-Rev.Med.Chem.. 2・6. 543-552 (2002)
• [文献書誌] H.Kakuta: "Fluorescent bioprobes for visualization of puromycin-sensitive aminopeptidase in living cells"Bioorg.Med.Chem.Lett.. 13・1. 83-86 (2002)
• [文献書誌] Y.Hashimoto: "Structural development of synthetic retinoids and thalidomide-related molecules"Cancer Chemother. Pharmacol.. (印刷中).
• [文献書誌] K.Nagasawa: "Total synthesisn of crambescidin 359"Org.Lett.. 4・2. 177-180 (2002)
• [文献書誌] H.Takahashi: "Novel Mannich-type nucleophilic substitution reaction with tertiary aromatic amines"Tetrahedron Lett.. 43・16. 2935-2938 (2002)
• [文献書誌] 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)
• [文献書誌] H.Takahashi: "Nucleophilic substitution on 4-hydroxymethylanilines under neutral conditions via aza quinone methide intermediate"Tetrahedron Lett.. 43・33. 5751-5753 (2002)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] T.Ishiwata: "Total synthesis of batazelladine D"Org.Lett.. 4・17. 2921-2924 (2002)
• [文献書誌] K.Nagasawa: "Stereoselective synthesis of trycyclic guanidine, the key component of the batzelladin alkaloids"Tetrahedron Lett.. 43・36. 6383-6385 (2002)
• [文献書誌] H.Kagechika: "Novel Synthetic Retinoids and Separation of the Pleiotropic Retinoidal Activities"Curr.Med.Chem.. 9・5. 591-608 (2002)
• [文献書誌] T.Imanishi: "BNAs : novel nucleic acid analogs with a bridged sugar moiety"Chem.Commun.(Camb).. 16. 1653-1659 (2002)
• [文献書誌] 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)
• [文献書誌] T.Imanishi: "Down-regulation of a gene-expression by an antisense BNA oligonucleotide"Nippon Yakurigaku Zasshi. 120・2. 85-90 (2002)
• [文献書誌] 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)
• [文献書誌] S.Obika: "Synthesis and conformation of 3',4'-BNA monomers, 3'-O, 4'-C methyleneribonucleosides"Tetrahedron. 58・15. 3093-3049 (2002)
• [文献書誌] Y.Hari: "Effecttive synthesis of C-nucleosides with 2',4'-BNA modification"Tetrahedron. 58・15. 3051-3063 (2002)
• [文献書誌] M.Yoshikawa: "Molecular and cytogenetic characterization of the mouse ATP-binding cassette transporter Abcg4"Gene. 293・1-2. 67-75 (2002)
• [文献書誌] H.Yabuuchi: "ABCC13, an unusual truncated ABC transporter, is highly expressed in fetal human liver"Biochem.Biophys.Res.Commun.. 299・3. 410-417 (2002)
• [文献書誌] 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).
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] M.Negishi: "Rho family GTPases as key regulators for neuronal network formation"J.Biochem.(Tokyo). 132・2. 157-166 (2002)
• [文献書誌] K.Nakamura: "The rostral raphe pallidus nucleus mediates pyrogenic transmission from the preoptic area"J.Neurosci.. 22・11. 4600-4610 (2002)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] Y.Hippo: "Global gene expression analysis of gastric cancer by oligonucleotide microarrays"Cancer Research. 62・1. 233-240 (2002)
• [文献書誌] A.Mukasa: "Distinction in gene expression profiles of oligodendrogliomas with and without alleic loss of 1p"Oncogene. 21・25. 3961-3968 (2002)
• [文献書誌] 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)
• [文献書誌] Y.Fujiwara: "Influence of polyunsaturated fatty acid on gene expression in HepG2 cells"J.Anteroscler.Thromb.. 9・3. 157-162 (2002)
• [文献書誌] J.M.Chong: "Interleukin-1 beta Expression in Human Gastric Carcinoma with Epstein-Barr Virus Infection"J.Virol.. 76・13. 6825-6831 (2002)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] T.Nakamura: "Discrimination between phosphotyrosine-mediated signaling properties of conventional and neuronal Shc adapter molecules"Oncogene. 21・1. 22-31 (2002)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] H.Tsukada: "Comparative effects of methamphetamine and nicotine on the striatal [^<11>C]raclopride binding in unanesthetized monkeys"Synapse. 45・4. 207-212 (2002)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)