1996 年度実績報告書

真核細胞の細胞周期に作用する低分子プロープ開発研究

研究課題

研究課題/領域番号	07660128
研究機関	理化学研究所
研究代表者	長田裕之理化学研究所, 抗生物質研究室, 主任研究員 (80160836)
キーワード	細胞周期 / タキソ-ル / cdc2キナーゼ / 阻害剤 / トリプロスタチン / アセトフタリジン
研究概要	細胞周期とは、遺伝子の本体であるDNAを合成して2つの娘細胞へ分配するという一連の過程をさす。最近、細胞周期の調節機構が分子レベルで解明されるにしたがって、その制御機構に異常が生じ無秩序に増殖する状態が癌であると理解されるようになった。細胞周期を阻害する薬剤としてタキソ-ルやカンプトテシンが発見され、新しい抗癌剤として期待されている。そこで、我々は微生物代謝産物より新しい細胞周期阻害剤の探索を行った。バイオアッセイ系には、マウス乳癌FM3A細胞から樹立したp34cdc2キナーゼ温度感受性変異株tsFT210細胞を用いた。本細胞は、39°Cでcdc2キナーゼの酵素活性が失活するためにG2期で細胞周期の進行を停止し、32°Cに戻すと再び細胞周期の進行を開始した。この条件下で、微生物培養サンプルを細胞培養液に添加し、細胞周期の停止作用をフローサイトメーターで測定した。スクリーニングの結果、Penicillium sp BM923株が生産するアセトフタリジンが細胞周期のM期で阻害することを見出した。アセトフタリジンは水溶液中で不安定であるため、まず不活性前躯体のヒドロキシメレインを単離し、それを酸性下加熱処理(変換)してアセトフタリジンに導いた。さらに糸状菌Aspergillus fumigatusが、新規なジケトピペラジン系化合物(トリプロスタチン、スピロトリプロスタチン、シクロトリプロスタチンなど)を生産することを明らかにした。トリプロスタチンA、Bはそれぞれ50μg/ml、12.5μg/mlで細胞周期を完全にM期で停止させた。さらにトリプロスタチンは、in vitro系でチューブリンの重合を阻害することを明らかにした。

研究成果
(17件)

すべてその他

すべて文献書誌 (17件)

[文献書誌] Ubukata,M.,et al: "Sparoxomycine A1 and A2. new inducers of the flat reversion of NRK cells transformed by temperature sensitive rous sarcoma virus II. Isolation. ……" J.Antibiot.49. 65-70 (1996)
[文献書誌] Nishiyama,U.,et al: "Structure-activity relationship within a series of degradation products of tautomycin." Biosci.Biotech.Biochem.60. 103-107 (1996)
[文献書誌] Cui,C,B.,et al: "Acetophthalidin,a novel inhibitor of mamrualian cell cycle,produced by a fungus isolated from a sea sediment." J.Antibiot.49. 216-219 (1996)
[文献書誌] Isogai,Y.,et al: "Direct measurement of oscillatory generation of superoxide anions by single phagocytes." FEBS Lett.380. 263-266 (1996)
[文献書誌] Matsuura,N.,et al: "Morphology reversion activity of phosmidosine and phosmidosine B. a newly isolated derivative. on srctransformed NRK cells." J.Antibiot.49. 361-365 (1996)
[文献書誌] Cui,C,-B.,et al: "Novel mammalian cell cycle inhibitors. tryprostatins A. B and other diketopiperazines produced by Aspergillus fumigatus I. Taxonomy. ……" J.Antibiot.49. 527-533 (1996)
[文献書誌] Cui,C,-B.,et al: "Novel mammalian cell cycle inhibitors. tryprostatins A.B and other diketopiperazines produced by Asoergillus fumigatus II. Physico-chemical properties and structure." J.Antibiot.49. 534-550 (1996)
[文献書誌] Cui,Z.,et al: "The multidrug resistance-associated protein (MRP) subfamily (Yrsi/Yorl) of saccharomyces cerevisiae is important for the tolerance to a broad rang of organic anions." J.Biol.Chem.271. 14712-14716 (1996)
[文献書誌] Cui,C,-B.,et al: "Spirotryprostatin B. a novel mammalian cell cycle inhibitor produced by aspergillus fumigatus." J.Antibiot.49. 832-835 (1996)
[文献書誌] Shimizu,T.,et al: "Synthetic studies on reveromycin A : stereoselective synthsis of the spiroketal system." Tetrahedron Lett.37. 6755-6758 (1996)
[文献書誌] Cui,C,-B.,et al: "Novel mammalian cell cycle inhibitors,spirotryprostatins A and B. produced by Aspergillus Lumigatus,which inhibit mammalian cell cycle at G2/M Phase." Tetrahedron.52. 12651-12666 (1996)
[文献書誌] Ubukata,M.,et al: "Sparoxomycins A1 and A2. new inducers of the flat reversion of NRK cells transformed by temperature sensitive rous sarcoma virus. I Taxonomy. ……" J.Antibiot.49. 1096-1100 (1996)
[文献書誌] Sudo,T.,et al: "Isolation and characterization of the gene encoding an aminopeptidase involved in the selective toxicity of ascamycin toward Xanthomonas campestris pv. citri." Biochem.J.319. 99-102 (1996)
[文献書誌] Sodeoka,M.,et al: "Asymmetric synthesis of RK-682 and its analogs,and evaluation of their protein phosphatase inhibitory activities." Tetrahedron Lett.37. 8775-8778 (1996)
[文献書誌] Cui,C.B.,et al: "Novel mammalian cell cycle inhibitors. cyclotryprostatins A-D. produced by aspergillus fumigatus,which inhibit mammalian cell cycle at G2/M phase." Tetrahedron. 53. 59-72 (1997)
[文献書誌] Osada,H.,et al: "Screening of cell cycle inhibitors from microbial metabolites by a bioassay using a mouse cdc2 mutant cell line,tsFT210." Bioorg.Med.Chem.5. 193-203 (1997)
[文献書誌] 長田裕之(分担): "分子腫瘍学「細胞周期と細胞内シグナル伝達の阻害剤の項目」" 中外医学社:佐藤昇志、菊地浩吉編, 401-413 (1996)

1996 年度 実績報告書

真核細胞の細胞周期に作用する低分子プロープ開発研究

研究代表者

長田 裕之 理化学研究所, 抗生物質研究室, 主任研究員 (80160836)

研究成果

[文献書誌] Ubukata,M.,et al: "Sparoxomycine A1 and A2. new inducers of the flat reversion of NRK cells transformed by temperature sensitive rous sarcoma virus II. Isolation. ……" J.Antibiot.49. 65-70 (1996)

[文献書誌] Nishiyama,U.,et al: "Structure-activity relationship within a series of degradation products of tautomycin." Biosci.Biotech.Biochem.60. 103-107 (1996)

[文献書誌] Cui,C,B.,et al: "Acetophthalidin,a novel inhibitor of mamrualian cell cycle,produced by a fungus isolated from a sea sediment." J.Antibiot.49. 216-219 (1996)

[文献書誌] Isogai,Y.,et al: "Direct measurement of oscillatory generation of superoxide anions by single phagocytes." FEBS Lett.380. 263-266 (1996)

[文献書誌] Matsuura,N.,et al: "Morphology reversion activity of phosmidosine and phosmidosine B. a newly isolated derivative. on srctransformed NRK cells." J.Antibiot.49. 361-365 (1996)

[文献書誌] Cui,C,-B.,et al: "Novel mammalian cell cycle inhibitors. tryprostatins A. B and other diketopiperazines produced by Aspergillus fumigatus I. Taxonomy. ……" J.Antibiot.49. 527-533 (1996)

[文献書誌] Cui,C,-B.,et al: "Novel mammalian cell cycle inhibitors. tryprostatins A.B and other diketopiperazines produced by Asoergillus fumigatus II. Physico-chemical properties and structure." J.Antibiot.49. 534-550 (1996)

[文献書誌] Cui,Z.,et al: "The multidrug resistance-associated protein (MRP) subfamily (Yrsi/Yorl) of saccharomyces cerevisiae is important for the tolerance to a broad rang of organic anions." J.Biol.Chem.271. 14712-14716 (1996)

[文献書誌] Cui,C,-B.,et al: "Spirotryprostatin B. a novel mammalian cell cycle inhibitor produced by aspergillus fumigatus." J.Antibiot.49. 832-835 (1996)

[文献書誌] Shimizu,T.,et al: "Synthetic studies on reveromycin A : stereoselective synthsis of the spiroketal system." Tetrahedron Lett.37. 6755-6758 (1996)

[文献書誌] Cui,C,-B.,et al: "Novel mammalian cell cycle inhibitors,spirotryprostatins A and B. produced by Aspergillus Lumigatus,which inhibit mammalian cell cycle at G2/M Phase." Tetrahedron.52. 12651-12666 (1996)

[文献書誌] Ubukata,M.,et al: "Sparoxomycins A1 and A2. new inducers of the flat reversion of NRK cells transformed by temperature sensitive rous sarcoma virus. I Taxonomy. ……" J.Antibiot.49. 1096-1100 (1996)

[文献書誌] Sudo,T.,et al: "Isolation and characterization of the gene encoding an aminopeptidase involved in the selective toxicity of ascamycin toward Xanthomonas campestris pv. citri." Biochem.J.319. 99-102 (1996)

[文献書誌] Sodeoka,M.,et al: "Asymmetric synthesis of RK-682 and its analogs,and evaluation of their protein phosphatase inhibitory activities." Tetrahedron Lett.37. 8775-8778 (1996)

[文献書誌] Cui,C.B.,et al: "Novel mammalian cell cycle inhibitors. cyclotryprostatins A-D. produced by aspergillus fumigatus,which inhibit mammalian cell cycle at G2/M phase." Tetrahedron. 53. 59-72 (1997)

[文献書誌] Osada,H.,et al: "Screening of cell cycle inhibitors from microbial metabolites by a bioassay using a mouse cdc2 mutant cell line,tsFT210." Bioorg.Med.Chem.5. 193-203 (1997)

[文献書誌] 長田裕之(分担): "分子腫瘍学「細胞周期と細胞内シグナル伝達の阻害剤の項目」" 中外医学社:佐藤昇志、菊地浩吉編, 401-413 (1996)

1996 年度実績報告書

長田裕之理化学研究所, 抗生物質研究室, 主任研究員 (80160836)