Tosso Perrer N, Kong Yali, Scher Lauren, Cummins Ryan, Schneider Jeffrey, Rahim Said, Holman K Travis, Toretsky Jeffrey, Wang Kan, Üren Aykut, Brown Milton L
Center for Drug Discovery, Georgetown University Medical Center , New Research Building EP07, 3970 Reservoir Road, NW, Washington, D.C. 20057, United States.
J Med Chem. 2014 Dec 26;57(24):10290-303. doi: 10.1021/jm501372p. Epub 2014 Dec 12.
EWS-FLI1 is an oncogenic fusion protein implicated in the development of Ewing's sarcoma family tumors (ESFT). Using our previously reported lead compound 2 (YK-4-279), we designed and synthesized a focused library of analogues. The functional inhibition of the analogues was measured by an EWS-FLI1/NR0B1 reporter luciferase assay and a paired cell screening approach measuring effects on growth inhibition for human cells containing EWS-FLI1 (TC32 and TC71) and control PANC1 cell lines devoid of the oncoprotein. Our data revealed that substitution of electron donating groups at the para-position on the phenyl ring was the most favorable for inhibition of EWS-FLI1 by analogs of 2. Compound 9u (with a dimethylamino substitution) was the most active inhibitor with GI50 = 0.26 ± 0.1 μM. Further, a correlation of growth inhibition (EWS-FLI1 expressing TC32 cells) and the luciferase reporter activity was established (R(2) = 0.84). Finally, we designed and synthesized a biotinylated analogue and determined the binding affinity for recombinant EWS-FLI1 (Kd = 4.8 ± 2.6 μM).
EWS-FLI1是一种致癌融合蛋白，与尤因肉瘤家族肿瘤（ESFT）的发生有关。利用我们之前报道的先导化合物2（YK-4-279），我们设计并合成了一个类似物的聚焦文库。通过EWS-FLI1/NR0B1报告荧光素酶测定法和配对细胞筛选方法来测量类似物的功能抑制作用，该配对细胞筛选方法用于测量对含有EWS-FLI1的人类细胞（TC32和TC71）和不含该癌蛋白的对照PANC1细胞系的生长抑制作用。我们的数据表明，在苯环对位上取代供电子基团对类似物抑制EWS-FLI1最为有利。化合物9u（具有二甲氨基取代）是活性最高的抑制剂，GI50 = 0.26±0.1μM。此外，还建立了生长抑制（表达EWS-FLI1的TC32细胞）与荧光素酶报告活性之间的相关性（R(2) = 0.84）。最后，我们设计并合成了一种生物素化类似物，并测定了其与重组EWS-FLI1的结合亲和力（Kd = 4.8±2.6μM）。
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