發(fā)布時間：2018-11-02 12:26

【摘要】：采用熒光偏振高通量篩選的方法進行PLK1 PBD小分子抑制劑的篩選,對篩選出的陽性化合物F083-0063進行體外抗腫瘤活性研究,以期為尋找抗腫瘤藥物提供先導(dǎo)化合物。模型篩選獲取一個對PLK1 PBD抑制率較高的化合物F083-0063,其在10μg·mL~(-1)時的抑制率達(dá)(99.7±0.4)%;利用軟件Graphpad Prism 5計算IC_(50)為1.9±0.1μmol·L~(-1);噻唑藍(lán)比色法(MTT)研究該化合物對不同細(xì)胞系增殖的影響,結(jié)果顯示F083-0063能抑制多種腫瘤細(xì)胞系的增殖;流式細(xì)胞儀檢測發(fā)現(xiàn),其能促進細(xì)胞凋亡且能導(dǎo)致細(xì)胞G2/M期阻滯;劃痕實驗測定F083-0063對細(xì)胞遷移的影響,結(jié)果顯示其能抑制He La細(xì)胞遷移,在20μmol·L~(-1)時,遷移率低至(37.6±0.7)%。利用分子連接技術(shù)探討化合物與PLK1 PBD結(jié)構(gòu)域的親和力,發(fā)現(xiàn)F083-0063與PLK1 PBD有較好的親和性;免疫印跡法(Western blotting)檢測顯示F083-0063可以引發(fā)周期相關(guān)蛋白表達(dá)的增加。綜上所述,化合物F083-0063有明顯的抗腫瘤活性,并有望成為靶向PLK1 PBD的抗腫瘤先導(dǎo)化合物。
[Abstract]:PLK1 PBD small molecular inhibitors were screened by fluorescence polarization high throughput screening method. The antitumor activity of the selected positive compound F083-0063 was studied in vitro in order to provide a lead compound for searching for antitumor drugs. A compound F083-0063 with high inhibition rate to PLK1 PBD was obtained by model screening, and the inhibition rate of F083-0063was (99.7 鹵0.4)% at 10 渭 g mL~ (-1), IC_ (50) was 1.9 鹵0.1 渭 mol L ~ (-1) calculated by the software Graphpad Prism 5, and the inhibition rate of F083-0063 was (99.7 鹵0.4)% at 10 渭 g mL~ (-1). The effect of F083-0063 on the proliferation of different cell lines was studied by (MTT). The results showed that F083-0063 could inhibit the proliferation of various tumor cell lines. Flow cytometry showed that it could promote cell apoptosis and induce G 2 / M phase arrest. The effect of F083-0063 on cell migration was determined by scratch test. The results showed that F083-0063 could inhibit the migration of He La cells, and the mobility was as low as (37.6 鹵0.7)% at 20 渭 mol L ~ (-1). The affinity of F083-0063 to PLK1 PBD domain was studied by molecular binding technique. F083-0063 had good affinity with PLK1 PBD, and F083-0063 could induce the increase of cyclin-related protein expression by Western blot (Western blotting) assay. In conclusion, compound F083-0063 has obvious antitumor activity and is expected to be a leading antitumor compound targeting PLK1 PBD.
【作者單位】： 黑龍江中醫(yī)藥大學(xué);中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院醫(yī)藥生物技術(shù)研究所;
【基金】：國家自然科學(xué)基金面上項目資助(81370087)
【分類號】：R96;R94
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本文編號：2306011