目的 寻找以成纤维细胞生长因子受体1(FGFR1)为靶点, 并具有良好抗肿瘤活性的小分子抑制剂。方法 通过改造单羰基姜黄素类似物3,5-双(2-氟苯亚甲基)哌啶-4-酮(EF24), 合成了其类似物[3,5-双(2-溴苯亚甲基)哌啶-4-酮, B6];采用FGFR1酪氨酸激酶实验验证其靶向性;MTT法检测EF24和B6对正常人肝细胞HL7702、4种肿瘤细胞(人大细胞肺癌NCI-H460、人胃癌细胞SGC-7901、人小细胞肺癌A549、人星形胶质瘤细胞株U251)增殖的影响;Western blotting法检测B6(2.5、5、10 μmol/L)对NCI-H460细胞bFGF/FGFR下游信号蛋白表达的影响;Caspase-3试剂盒检测Caspase-3因子的表达。结果 EF24类似物B6可以以FGFR1为靶点, 抑制NCI-H460细胞FGFR1以及下游信号蛋白丝氨酸/苏氨酸蛋白激酶(AKT)和细胞外调节蛋白(ERK1/2)的磷酸化, 抑制肿瘤细胞的增殖, 促进细胞凋亡。结论 成功改造并获得靶向FGFR1的EF24类似物B6, 其具有良好的体外抗肿瘤活性, 为寻找FGFR1抑制剂候选抗肿瘤药物提供新思路。

Objective To search for the small molecule inhibitor with anti-tumor activity by fibroblast growth factor receptor 1 (FGFR1) as target. Methods The analogue B6 of EF24 was obtained by reforming mono carbonyl curcumin analogues and applied to identifying the target with FGFR1 kinase activity assay. The effect of EF24 and its analogue B6 on the proliferation of HL7702 in normal human and four tumor cells, such as NCI-H460, SGC-7901, A549, and U251; With the concentration of 2.5, 5, and 10 μmol/L, B6 was used to investigate the phosphorylation inhibition of bFGF/FGFR downstream signal protein expression in NCI-H460 cells and Caspase-3 factor expression. Results With the FGFR1 as target, B6 could inhibit the phosphorylation of FGFR1 in NCI-H460 cells, AKT, and ERK1/2; It also could inhibit the proliferation of cancer cells and promote cell apoptosis. Conclusion The analogue B6 of EF24 is obtained from the leading compound EF24 with in vitro anti-tumor effect, which provides the basis of looking for the candidate anti-tumor drugs of FGFR1 inhibitor.

“十二五”国家科技支撑计划项目(2011BAI04B04);浙江省自然科学基金资助项目(LY15H280014)