Development of inhibitors for polyamine metabolism including cell death

开发多胺代谢（包括细胞死亡）抑制剂

基本信息

批准号：
12672158
负责人：
SHIRAHATA Akira
金额：
$ 2.5万
依托单位：
Josai University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

Aiming at elucidation of the mechanism of cell death accompanied by the polyamine accumulation in cells, the following research has been done to develop strong inhibitor for spermidine/spermine-N^1-acetyltransferase (SSAT): 1) Development of novel assay method for SSAT and screening assay method for compounds inhibiting SSAT activity, 2) Synthesis of polyamine analogues and examination of their effect on cell growth or apoptosis in cultured cell lines, 3) Changes in cell volume and polyamine distribution in cells accumulated with polyamine.It was found that N1-dansylnorspermine was very useful as a fluorescent substrate for non-radio isotopic SSAT assay method using recombinant human SSAT. The kinetic studies were performed for the recombinant SSAT protein and cellular SSAT activity in HTC cells. It was shown that the method was useful for measuring the activity in biological sample having low level of the enzyme. From the screening for the inhibitor, we found that bis(alkyl)norspermidine was a very potent one, whose IC50 to SSAT activity was much lower than bis(ethyl)spermine, and had inhibition activity for polyamine oxidase.It was also found that the compounds synthesized in this study were effective for cell growth inhibition or induction of apoptosis.The changes in cell volume and distribution of polyamine were examined by Coulter counter and ultrafiltration method, respectively, for the cells accumulated with spermidine. It was suggested that the cell volume was decreased and the distribution of polyamine was changed by the accumulation of cellular polyamine.

Aiming at elucidation of the mechanism of cell death accompanied by the polyamine accumulation in cells, the following research has been done to develop strong inhibitor for spermidine/spermine-N^1-acetyltransferase (SSAT): 1) Development of novel assay method for SSAT and screening assay method for compounds inhibiting SSAT activity, 2) Synthesis of polyamine analogues and examination of their effect on cell growth or培养的细胞系中的凋亡，3）细胞体积和多胺积聚的细胞中多胺分布的变化。它发现N1-丹尼斯林植物是使用重组人类SSAT的非拉迪奥同位素SSAT分析方法的荧光底物非常有用。对HTC细胞中的重组SSAT蛋白和细胞SSAT活性进行了动力学研究。结果表明，该方法对于测量具有低酶水平的生物样品的活性很有用。从抑制剂的筛选中，我们发现双（烷基）杂交是一种非常有效的抑制剂，其IC50至SSAT活性远低于BIS（乙基）的精子，并且具有多胺氧化酶的抑制活性。它也发现了该研究中的化合物在细胞生长抑制或指标中的分布及其分布的拟南条及其分布，并且是多型的变化。分别用于用精子积聚的细胞的Coulter计数器和超滤法。有人提出，细胞体积减少，多胺通过细胞多胺的积累改变了多胺的分布。