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目的:以酵母细胞为模式菌高通量筛选具有抗肿瘤活性的Aurora-B激酶抑制药。方法:以野生型酵母菌Y300和ipll-321温度敏感型突变株为模式菌,筛选Aurora-B激酶抑制药;体外酶学实验验证候选化合物对Aurora-B激酶的抑制作用;MTT方法检测阳性化合物对肿瘤细胞的杀伤活性;流式细胞术检测阳性化合物对肿瘤细胞的周期阻滞。结果:经筛选得到12个化合物和5个微生物发酵液;其中浓度在10μg·ml~(-1)时对重组纯化的人Aurora-B激酶活性抑制率高于50%的有7个,其中3个活性较好的化合物IC_(50)分别为10.253,1.826和2.054μmol·L~(-1);其中3个化合物对Hela细胞的IC_(50)分别为4.255,15.326和1.032μmol·L~(-1),对HepG2细胞的IC_(50)分别为5.387,17.465和1.725μmol·L~(-1),对HCT116细胞的IC_(50)分别为5.380,8.528和2.029μm o1.L~(-1)。11号化合物能够诱导HCT116细胞G2/M期阻滞。结论:以Y300和ipl1-321酵母细胞为高通量筛选模型筛选到多个新的具有抗肿瘤活性的Aurora-B激酶抑制药。
OBJECTIVE: To screen the Aurora-B kinase inhibitor with anti-tumor activity using yeast cells as model fungus high-throughput. Methods: Aurora-B kinase inhibitors were screened by the wild-type yeast Y300 and ipll-321 temperature-sensitive mutant strains. The inhibitory effect of the candidate compounds on Aurora-B kinase was verified by in vitro enzymatic experiments. The positive compounds Cytotoxicity to tumor cells was determined by flow cytometry. Results: Twelve compounds and five microbial fermentation broths were screened out. Among them, seven of them were inhibited when the concentration of Aurora-B kinase was 10μg · ml ~ (-1) The IC 50 of the active compounds were 10.253, 1.826 and 2.054μmol·L -1, respectively. The IC 50 values of three compounds against Hela cells were 4.255, 15.326 and 1.032μmol·L -1 -1). The IC 50 of HepG2 cells were 5.387, 17.465 and 1.725 μmol·L -1, respectively. The IC 50 of HCT116 cells were 5.380, 8.528 and 2.029 μm, 1). Compound No. 11 can induce G2 / M arrest in HCT116 cells. Conclusion: Several new anti-tumor Aurora-B kinase inhibitors were screened by using Y300 and ipl1-321 yeast cells as a high-throughput screening model.