基于药效团和分子对接的FLT3抑制剂筛选方法

Pharmacophore modeling and molecular docking based method for screening FLT3 inhibitors

目的建立虚拟筛选与生物检测相结合的方法来筛选FLT3抑制剂。方法建立了基于药效团和对接的虚拟筛选方法用于筛选靶向FLT3的化合物,并建立FLT3-ITD过表达的细胞模型进一步验证抑制剂的药效及作用机制。结果通过筛选商业化合物库,成功发现了1个含有3-苯基吡唑并[1,5-a]嘧啶骨架的化合物(命名74),可以选择性地抑制表达FLT3-ITD的BaF3细胞和AML细胞增殖。进一步研究表明,在细胞实验中,74也能克服FLT3-ITD-D835V、-D835Y、-F691L、-N676K等二次突变。机制研究表明,74通过抑制FLT3介导的通路抑制AML细胞的生长。结论74是1个潜在的抑制FLT3-ITD的苗头分子,它的发现验证了FLT3筛选方法的准确性,并可用来寻找海洋来源的FLT3抑制剂。

Objective To develop the method of virtual screening coupled with biological detection for screening FLT3 inhibitors.Methods The pharmacophore-and docking-based screening methods were developed for virtualy screening the hits targeting FLT3,and FLT3-ITD overexpressing cellular models for further verifying the inhibitory effect and its related mechanism.Results By screening the commercial library,a compound harboring scaffold of 3-phenylpyrazolo[1,5-a]pyrimidine,namely 74 was successfully discovered,which selectively inhibited the proliferation of FLT3-ITD expressing BaF3 cells and AML cells.Further study indicated that compound 74 could also overcome secondary mutations,including FLT3-ITD-D835V,-D835Y,-F691L,-N676K mutations in cellular assays.Mechanically,compound 74 inhibited the growth of AML cells by suppressing FLT3 mediated pathway.Conclusion The results indicated that compound 74 was a promising hit for FLT3-ITD inhibition.The discovery of compound 74 validated the screening methods,which could be further used to find marine-derived FLT3 inhibitors.