微生物来源的Aurora-B激酶抑制剂的分离、结构鉴定和抗肿瘤活性研究  

Isolation,structure identification and anti-tumor activity of potential small-molecular inhibitor of human Aurora-B kinase from microbial nature products

作　　者：牛玉强[1] 姜威[1] 余利岩[1] 王彦昶[1] 李妍[1] 司书毅[1] 

机构地区：[1]中国医学科学院医药生物技术研究所,北京100050

出　　处：《中国医药生物技术》2011年第2期116-120,共5页Chinese Medicinal Biotechnology

摘　　要：目的从微生物代谢产物中分离和纯化Aurora-B激酶抑制剂并检测其抗肿瘤活性。方法以野生型酵母菌Y300和ipl1-321温度敏感型突变株为模式菌跟踪活性组分;从阳性放线菌I07A-01038发酵产物中分离纯化活性化合物并进行结构鉴定;体外酶学实验验证阳性化合物对Aurora-B激酶的抑制活性;MTT法检测阳性化合物对肿瘤细胞增殖的抑制活性;AnnexinV-FITC/PI双染法检测活性化合物对肿瘤细胞早期凋亡的诱导作用。结果从阳性放线菌I07A-01038发酵产物中得到活性化合物酒渣碱甲酯(flazinmethylester),酒渣碱甲酯对ipl1-321突变株具有特异性抑制作用,其在野生型酵母菌株Y300和突变株ipl1-321的IC50分别为48μmol/L和24μmol/L;体外酶学实验证实其对Aurora-B激酶具有抑制作用,其IC50为10μmol/L(ATP=50μmol/L);酒渣碱甲酯对HepG2、A549、Hela肿瘤细胞均具有杀伤活性,IC50分别为13、11和10μmol/L。并能够诱导Hela细胞发生早期凋亡。结论得到一个微生物来源的具有抗肿瘤活性的Aurora-B激酶抑制剂—酒渣碱甲酯。Objective To find potential small-molecular Aurora-B kinase inhibitors from microbial metabolites and research on anti-cancer activity. Methods In vitro biochemical assay was performed to test the inhibition of candidates determined by screening with wild-type yeast strain and ipll-321 temperature sensitive strain. MTT assay was used to evaluate the inhibitory effect of flazin methyl ester on the proliferation of tumor cells. Annexin V-FITC and PI were performed to determine the induction of flazin methyl ester on the early apoptosis of tumor cells. Results The compound 70-4 named flazin methyl ester was isolated and purified from the fermentation products of a positive strain 107A-01038. The flazin methyl ester inhibited the growth of ipll-321 temperature sensitive mutant more dramatically than the wild-type yeast cells, and its MIC was 24μmo//L and 48 μmol/L. The flazin methyl ester inhibited Aurora-B kinase activity in vitro, and its IC50 was 10 ~tmol/L when the ATP concentration was 50 μmol/L. The flazin methyl ester inhibited the growth of tumor cells, such as HepG2, A549 and Hela, and its IC50 was 13, 11 and 10 μmol/L respectively. The flazin methyl ester could induce early apoptosis of tumor cells, Hela. Conclusion A potential small-molecular inhibitor of human Aurora-B kinase, flazin methyl ester, was obtained.

关 键 词：磷酸转移酶类 细胞抑制剂 细胞增殖 细胞凋亡 

分 类 号：R91[医药卫生—药学] R96