放线菌KS-5555产生的抑制VCAM-1表达的活性物质的研究

Studies on active compounds which prevent the transcriptional activity of VCAM1 from actinomyces KS5555

使用一个含有人ＶＣＡＭ－１基因启动子和荧光素酶编码基因的转染细胞株，从微生物代谢产物中筛选能够抑制由ＴＮＦ－α诱导的ＶＣＡＭ－１表达的活性物质。在筛选过程中，应用有机溶媒萃取、ＯＤＳ柱层析及ＨＰＬＣ等方法，从一株放线菌ＫＳ－５５５５的发酵液中分离到了ＫＳ－５５５５－Ｅ、Ｆ、Ｇ３种生物活性物质。它们对与人ＶＣＡＭ－１基因的启动子相连接的荧光素酶基因的表达显示出较强的抑制活性，其ＩＣ５０值分别为０．３２、０．１４、０．１９μｍｏｌ／Ｌ。经各种理化特性及ＵＶ、ＭＳ、ＨＰＬＣ等分析，确定化合物ＫＳ－５５５５－Ｅ、Ｆ、Ｇ分别与脂酰ＣｏＡ合成抑制剂ｔｒｉａｃｓｉｎＣ、Ｄ、Ａ的结构相同。

We used a stably transfected cell line carrying the promoter region of human VCAM1 gene fused to the coding region of firefly luciferase gene to screen samples that prevent the TNFinduced stimulation of VCAM1 transcription from microbial metabolites.In the course of screening, three physiologically compounds KS5555E,F,G were isolated from actinomyces KS5555 by solvent extraction, ODS column chromatography and HPLC etc. These compounds showed strong inhibitory activity for the transcription of firefly luciferase gene fused to the promoter region of the human VCAM1 gene, the values of IC50were 0.32, 0.14, 0.19μmol/L respectively. The structures of KS5555 E,F,G were elucidated to be identical with the acylCoA synthetase inhibitor triacsin C,D,A by means of physicochemical properties and UV, MS spectra and HPLC analysis.