论文部分内容阅读
目的组织蛋白酶B(cathepsin B,CatB)是一种主要存在于溶酶体的半胱氨酸蛋白水解酶,具有广谱蛋白水解酶活性,能够降解多种基底膜蛋白,为癌细胞的转移打开通道,促进肿瘤细胞向深部组织浸润。因此,研究CatB抑制剂在抗肿瘤转移方面具有潜在的应用价值。本研究筛选CatB抑制剂,发现潜在的抗癌药物先导化合物。方法利用自建的快速、高效的组织蛋白酶B抑制剂的高通量筛选方法,从大量真菌菌库及其来源的代谢产物中筛选能够产生CatB抑制活性产物的菌株,对其发酵产物进行有机溶剂提取、硅胶柱色谱、Sephadex LH-20柱色谱及HPLC制备分离获得活性化合物,经紫外、质谱、核磁等理化数据的分析进行结构鉴定。结果筛选分离得到一个活性化合物F04ZA-1203A,结构解析确定了其为已知化合物thielavin B,该化合物对CatB的抑制IC50为1.85μg/ml。结论F04ZA-1203A(thielavin B)对CatB有强的抑制活性,国内外未见报道。
Objective Cathepsin B (CatB), a cysteine protease mainly present in lysosomes, possesses broad-spectrum proteolytic enzyme activity and is capable of degrading various basement membrane proteins and opening up for the metastasis of cancer cells Channel, promote the tumor cells to deep tissue infiltration. Therefore, the study of CatB inhibitors has potential value in antitumor metastasis. This study screened for CatB inhibitors and found potential anticancer drug lead compounds. Methods A high-throughput screening method of rapid and efficient cathepsin B inhibitors was developed to screen out the strains capable of producing CatB inhibitory activity from a large number of fungal strains and their metabolites. The fermentation products were subjected to organic solvent Extraction, silica gel column chromatography, Sephadex LH-20 column chromatography and HPLC preparation. The compounds were identified by UV, MS and NMR. Results The active compound F04ZA-1203A was screened and isolated. Its structural elucidation confirmed that it was a known compound thielavin B, and its IC50 against CatB was 1.85 μg / ml. Conclusion F04ZA-1203A (thielavin B) has a strong inhibitory activity on CatB, which has not been reported at home and abroad.