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構造、配列、代謝情報を基盤とする抗がん性核酸誘導体の設計

Research Project

Project/Area Number 12217001
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionHokkaido University

Principal Investigator

松田 彰  北海道大学, 大学院・薬学研究科, 教授 (90157313)

Project Period (FY) 2000 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥43,200,000 (Direct Cost: ¥43,200,000)
Fiscal Year 2004: ¥9,700,000 (Direct Cost: ¥9,700,000)
Fiscal Year 2003: ¥8,800,000 (Direct Cost: ¥8,800,000)
Fiscal Year 2002: ¥9,000,000 (Direct Cost: ¥9,000,000)
Fiscal Year 2001: ¥7,700,000 (Direct Cost: ¥7,700,000)
Fiscal Year 2000: ¥8,000,000 (Direct Cost: ¥8,000,000)
Keywords固形がん / 放射線 / 代謝拮抗剤 / ECyd / CNDAC / 併用効果 / survivin / Chk1 / UCK-2 / Bcl-2 / ネクローシス / アポトーシス / X線 / G2 / M期checkpoint / シスプラチン / HMG蛋白 / 4-チオリボヌクレオシド / ミトコンドリア / Bax / トランスロケーション / z-VAD-fmk / ウリジン・シチジンキナーゼ / 核酸代謝酵素 / デコイ核酸 / DNA(cytosine-5)methyltransferase / 4'-チオシチジン誘導体
Research Abstract

CNDACは、その三リン酸体がDNA中に取込まれると、β-脱離反応により一本鎖切断を起こすように分子設計した抗癌性ヌクレオシドである.本研究では、その機序解析を詳細に行い、以下の諸点を明らかにした.1)5種類の血液癌細胞、3種類の固形癌細胞および患者から採取した2種類の繊維芽細胞を用いてG2/M期の割合をCNDAC未処理細胞と処理細胞間で比較したところ、未処理細胞では14〜29%なのに対して処理細胞では33〜55%と有意に増加していた.2)G2期停止に伴って、Chk1のSer-317,Ser-345のリン酸化が起こり、さらにChk1の標的であるCdc25CのSer-216のリン酸化が進行する.その結果、Cdk1のTyr-15がリン酸化されたままになりCyclin B1レベルを上昇させG2期停止に至る.3)CNDAC処理細胞にUCN-01(Chk1阻害剤)を添加すると、G2期細胞の割合が急速に減少し、一時的にG1期細胞が増える.その後、G1期細胞が減少し、アポトーシスが開始される.UCN-01単独ではこのような作用は観察されないので、UCN-01はG2期停止をabrogateし、アポトーシスによる細胞死を大幅に増加させる.4)ML-1細胞をCNDAC処理したところ、二本鎖切断が主に起きていることが明らかになった.この効果もまた、UCN-01添加で増強された.以上のように、CNDACのG2期停止に至る機序の解明とそれに基づくChk1の阻害剤による作用増強が可能になった.
がん細胞にX線を照射するとDNAの二本鎖切断を起こすために、G2 checkpoint経路が活性化されG2期で停止し、DNAの修復を待つ.一方、ECydの代謝物であるECyd 5'-三リン酸はRNAポリメラーゼを強力に阻害し、これが引金となりアポトーシスを起こす.この両者を併用した場合に、X-線照射ではヒト胃がん細胞MKN45細胞はネクローシスしか起こさないのに対して、ECydを添加するとアポトーシスへと細胞死の方向を変えることが明らかになった.この機序を明らかにすべく種々検討の結果、ECyd処理により、抗アポトーシス蛋白の一種であり各種がん細胞に高率に発現しているsurvivinがmRNAレベルでも蛋白レベルでも急激に減少することが明らかになった.SurvivinはCdc2-cyclin B1によりリン酸化を受け、M期で抗アポトーシスを発揮しているが、ECyd処理によりsurvivinが合成されなくなったためG2 checkpointが解除され、caspase依存的なアポトーシスが誘導されたと考えられる.

Report

(5 results)
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • 2000 Annual Research Report

Research Products

(20 results)

All 2004 Other

All Journal Article Publications

  • [Journal Article] Antitumor activity of sugar-modified cytosine nucleosides.2004

    • Author(s)
      Matsuda, A., et al.
    • Journal Title

      Cancer Sci. 95

      Pages: 105-111

    • NAID

      10012710063

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Structural basis for the specificity, catalysis and regulation of human uridine-cytidine kinase.2004

    • Author(s)
      Suzuki, N.N., et al.
    • Journal Title

      Structure 12

      Pages: 751-764

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Synthesis and physical and physiological properties of 4'-thioRNA : Application to post-modification of RNA aptamer toward NF-κB.2004

    • Author(s)
      Hoshika, S., et al.
    • Journal Title

      Nucleic Acids Res. 32

      Pages: 3815-3825

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A crucial role of uridine/cytidinekinase2inantitumoractivity of 3'-ethynyl nucleosides.2004

    • Author(s)
      Murata, D., et al.
    • Journal Title

      Drug Metab.Dispos. 32

      Pages: 1178-1182

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A novel anticancer ribonucleoside, 1-(3-C-ethynyl-β-D-ribo-pentofuranosyl) cytosine, enhances radiation-induced cell death in tumor cells.2004

    • Author(s)
      Inanami, O., et al.
    • Journal Title

      Radiat.Res. 162

      Pages: 635-645

    • Related Report
      2004 Annual Research Report
  • [Publications] Nomura, M., Shuto, S., Matsuda, A.: "Synthesis of the cyclic and acyclic acetal derivatives of 1-(3-C-ethynyl-β-D-ribo-pentofuranosyl)cytosine, a potent antitumor nucleoside. Design of prodrugs to be selectively activated in tumor tissues via a bio-reduction hydrolysis mechanism."Bioorg.Med.Chem.. 11. 2453-2461 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Minakawa, N., Kato, Y., Uetake, K., Kaga, D., Matsuda, A.: "An improved large scale synthesis of 1,4-anhydro-4-thio-D-ribitol."Tetrahedron. 59. 1699-1702 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Sukeda, M., Ichikawa, S., Matsuda, A., Shuto, S.: "The first radical method for the introduction of an ethynyl group using a silicon tether and its application to the synthesis of 2-deoxy-2-C-ethynylnucleosides."J.Org.Chem.. 68. 3465-3475 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Maseoka, A., Matsubara, M., Hasegawa, R., Tanaka, T., Kuris, S., Terato, H., Ohyama, Y, Karino N., Matsuda, A., Ide, H.: "Mammalian 5-formyluracil-DNA glycosylase. 2. Role of SMUG1 uracil-DNA glycosylase in repair of 5-formyluracil and other oxidaized and deaminated base lesions."Biochemistry. 42. 5003-5012 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Minakawa, N., Kojima, N., Hikishima, S., Sasaki, T., Kiyosue, A., Atsumi, N., Ueno, Y., Matsuda, A.: "New base pairing motifs. The synthesis and thermal stability of oligodeoxynucleotides containing imidazopyridopyrimidine nucleosides with the ability to form four hydrogen bonds."J.Am.Chem.Soc.. 125. 9970-9982 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nomura, M., et al.: "Practical large-scale synthesis of 1-(3-C-ethynyl-b-D-ribo-pentofuranosyl)cytosine (ECyd), a potent antitumor nucleoside. Isobutyryloxy group as an efficient anomeric leaving group in the Vorbruggen glycosylation reaction"Tetrahedron. 58. 1279-1288 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shimamoto, Y., et al.: "Cellular and biochemical mechanisms of the resistance of human cancer cells to a new anticancer ribo-nucleoside, TAS-106"Jpn. J. Cancer Res.. 93. 445-452 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shimamoto, Y., et al.: "Sensitivity of human cancer cells to the new anticancer ribo-nucleoside TAS-106 is correlated with expression of uridine-cytidine kinase2"Jpn. J. Cancer Res.. 93. 825-833 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Obata, T., et al.: "Deletion mutants of human deoxycylidine kinase mRNA in cells resistant to antitumor cytosine nucleoside"Jpn.J.Cancer Res.. 92. 793-798 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Azuma, A., et al.: "Cellular pharmacokinetics and pharmacodynamics of the deoxycytidine analog 2'-C-cyano-2'-deoxy-1-β-D-arabino-pentofuranosylcytosine (CNDAC)"Biochem.Pharmacol.. 61. 1497-1507 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Shimamoto, Y., et al.: "Antitumor activity and pharmacokinetics of TAS-106, 1-(3-C-ethynyl-β-D-ribo-pentofuranosyl)cytosine"Jpn.J.Cancer Res.. 92. 343-351 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Y.Nomura, et al.: "2-Chloro-2'-deoxyadenosine induces apoptosis through the Fas/Fas ligand pathway in human leukemia cell line MOLT-4."Leukemia. 14. 299-306 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Naka, et al.: "The stereoselective synthesis of 4'-β-thioribonucleosides via the Pummerer reaction."J.Am.Chem.Soc.. 122. 7233-7243 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] M.Nomura, et al.: "Development of an efficient intermediate, α-[2-(trimethylsilyl) ethoxy]-2-N-[2-(trimethylsilyl) ethoxycarbonyl] folic acid, for the synthesis of folate (γ)-conjugates, and its application to the synthesis of folate-nucleoside conjugates."J.Org.Chem.. 65. 5016-5021 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Asai, et al.: "Targeting and antitumor efficacy of liposomal 5'-O-dipalmitoylphosphatidyl 2'-C-cyano-2'-deoxy-1-β-D-arabino-pentofuranosylcytosine in mice lun bearing Bl6BL6 melanoma."Cancer Lett.. 162. 49-56 (2001)

    • Related Report
      2000 Annual Research Report

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Published: 2000-03-31   Modified: 2018-03-28  

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