新型TRK激酶小分子抑制剂的筛选与验证

Screening and Verification of Small Molecular Inhibitors against Tropomyosin-Related Kinases(TRKs)

原肌球蛋白相关激酶(Tropomyosin-Related Kinase,TRK)属于受体酪氨酸激酶家族,具有调节细胞增殖、分化、凋亡、代谢等作用.在多种肿瘤细胞中发现TRK激酶编码基因NTRK的融合现象,TRK蛋白过表达或激酶活性组成性激活促进了肿瘤的发生发展.以TRK激酶为靶标的小分子抑制剂正处于研发之中,如Entrectinib等.本研究以Entrectinib为阳性对照,从小分子化合物库中筛选得到Crizotinib、LY2874455等7个新颖的TRK激酶小分子抑制剂,结构计算提示Dovitinib等4个化合物均属于ATP竞争性抑制剂.在NTRK1基因融合的KM12细胞体外增殖实验中,全部的TRK激酶抑制剂均能抑制细胞增殖,且LY2874455最为显著.在细胞的联合用药实验中,发现Crizotinib与Entrectinib的联用具有协同作用.

Tropomyosin-related kinases (TRKs) are a group of receptor tyrosine kinases, which regulate many cellular functions including cell proliferation, cell differentiation, metabolism and apoptosis. Gene fusion of neurotropic tropomyosin receptor kinase genes ( NTRK ) coding for TRKs contributes to constitutive activation or overexpression of TRKs, which increase the risk of tumor genesis. Up to now, a variety of TRK inhibitors including Entrectinib are under research and development. In this study, 7 novel inhibitors against TRKs including Crizotinib, LY2874455 are obtained from a HTRF assay, with Entrectinib as a positive control. Structure calculation demonstrates that Dovitinib and other 3 compounds are ATP competitive inhibitors. All the novel TRK inhibitors show strong suppressive effect on cell proliferative in the colorectal carcinoma cell line KM12 with NTRK1 gene fusion, with LY287445 as the most significant among them. Furthermore, the combination of Crizotinib and Entrectinib was demonstrated to have a synergistic effect on KM12 cell proliferation.