Isolation and mechanism study of cell cycle inhibitors from microbial metabolites.

微生物代谢产物中细胞周期抑制剂的分离及机制研究。

• 批准号: 09460058
• 负责人: OSADA Hiroyuki
• 金额: $ 10.05万
• 依托单位: RIKEN
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-09460058/
• 关键词: G1 phase; protein synthesis inhibition; cell cycle; Curvularia sp.; new compound; sesquiterpene; trichotecene; 細胞周期阻害剤; 微小管; チューブリン; ピロネチン; ビオチン標識; チユープリン; トリプロスタチン; サイトカラシン; サイクリン; テルペプチン; ホスミドシン

We have been screening microbial metabolites of soil actinomycetes and fungi to obtain new cell cycle inhibitors. The cell cycle analysis was carried out by flow cytometry using tsFT210 cell, a temperature-sensitive mutant cell line of cdc2 kinase. During this screening, a new cell cycle inhibitor named curvularol was discovered from the fermentation broth of a fungus, Curvularia sp. 97-F166, isolated in Nagasaki Prefecture. Curvularol was purified by a successive column chromatography and obtained as a colorless needle crystal. Its molecular weight and molecular formula were determined to be 266 and C_<15>H_<22>O_4, respectively, according to high-resolution FAB mass spectrometry. The structure of curvularol was elucidated by the detailed analyses of its 1D and 2D NMR spectra. It is a new sesquiterpen structurally related to trichotecenes to some extent.Curvularol showed a weak antimicrobial activity at the concentration of 100 μg/ml, but showed a strong cytotoxicity at the concentration range of 50〜500 ng/ml, in vitro. The effect of curvularol against the cellular macromolecular synthesis was examined, and the primary target of curvularol was suggested to be protein synthesis. It blocked the cell cycle progression at G1 phase of various cell lines including, tsFT210, src^<ts>-NRK at 50 ng/ml.

我们一直在筛选土壤放线菌和真菌的微生物代谢产物，以获得新的细胞周期抑制剂。通过使用TSFT210细胞流式细胞仪进行细胞周期分析，TSFT210细胞是CDC2激酶的温度敏感突变细胞系。在此筛选过程中，从真菌的发酵液中发现了一种新的细胞周期抑制剂。 97-F166，在长崎县隔离。通过成功的柱色谱法纯化曲线醇，并作为无色针晶体获得。根据高分辨率FAB质谱法，其分子量和分子公式分别确定为266和C_ <15> H_ <22> O_4。通过其1D和2D NMR光谱的详细分析来阐明曲线醇的结构。这是一种在一定程度上与毛滴虫结构相关的新的倍烯培养子。白菜在100μg/ml的浓度下显示出弱的抗菌活性，但在50-500 ng/ml的浓度范围内显示出强的细胞毒性，在体外。检查了曲线醇对细胞大分子合成的影响，并建议曲线醇的主要靶标是蛋白质合成。它阻止了各种细胞系的G1相的细胞周期进程，包括TSFT210，SRC^<ts> -NRK在50 ng/ml时。

项目成果

M.Watabe, et al: "Activation of MST/Krs and c-Jun N-terminal kinases by different signaling pathways during cytotrienin A-induced apoptosis."J.Biol.Chem.. 275. 8766-8771 (2000)

M.Watabe 等人：“在细胞三烯蛋白 A 诱导的细胞凋亡过程中，通过不同的信号传导途径激活 MST/Krs 和 c-Jun N 末端激酶。”J.Biol.Chem.. 275. 8766-8771 (2000)

Y.Honda, et al: "Isolation, and biological properties of a new cell cycle inhibitor, Curvularol, isolated from Curvularia sp.RK97-F166."J.Antibiot.. 54. 10-16 (2001)

Y.Honda 等人：“从 Curvularia sp.RK97-F166 中分离出的新型细胞周期抑制剂 Curvularol 的分离和生物学特性。”J.Antibiot.. 54. 10-16 (2001)

M.Maruyama, et al: "Immunolocalization of p38 MAP kinase in mouse brain."Brain Res.. 887. 350-358 (2000)

M.Maruyama 等人：“小鼠大脑中 p38 MAP 激酶的免疫定位。”Brain Res.. 887. 350-358 (2000)

渡部正彦 他: "Effect of microgravity on cell proliferation signal."日本マイクログラビティ応用学会誌. 17. 87-90 (2000)

Masahiko Watanabe 等人：“微重力对细胞增殖信号的影响。”日本微重力应用学会杂志 17. 87-90 (2000)。

Cui C.B.: "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)

Cui C.B.：“新型哺乳动物细胞周期抑制剂，环胰前列腺素A-D，由烟曲霉产生，可抑制哺乳动物细胞周期G2/M期”四面体。