白细胞弹性蛋白酶抑制剂筛选模型的建立及抑制剂F02ZA-2554A的研究

Studies on the high throughput screening assay for HLE inhibitor and an inhibitor F02ZA-2554A

利用人源的白细胞弹性蛋白酶建立了一种快速、微量、灵敏的高通量体外筛选方法。利用该模型从微生物菌种库菌株的次生代谢产物中筛选得到一阳性真菌F02ZA-2554。该菌株发酵液经有机溶剂提取、硅胶柱层析、及HPLC ODS反相柱层析等活性跟踪下的分离纯化,得到活性化合物F02ZA-2554 A,该化合物对白细胞弹性蛋白酶有中等强度的抑制活性,其IC50为32.4μmol/L。通过对该化合物的紫外、质谱、核磁等理化分析,确定该化合物为蒽醌类化合物中的大黄素甲醚(physcion),该化合物的抗炎活性属首次报道。

Using L-pyroglutamyl-L-prolyl-L-valine-p-NA as the substrate, a high throughput screening assay was developed for screening HLE inhibitors with high S/N ratios （〉5） and Z＇-factors （〉0. 85）. In screening for HI.E inhibitors from metabolites of microorganisms, a compound named F02ZA-2554A was isolated from fungal broth with IC50 of 32.4μmol/L. The spectrum data （UV, IR, MS and NMR） showed that it is physcion, which is the ubiquitous component in many kinds of herbs. These results maybe revealed the mechanism of physcion on anti-inflammation, which has never been reported before.