Focal adhesion kinase (FAK) is an intracellular non-receptor tyrosine kinase responsible for development of various tumor types. Aiming to explore new potent inhibitors, two series of 2,4-disubstituted-7H-pyrrolo[2,3-d]pyrimidine derivatives were designed and synthesized on the base of structure-based design strategy. Biological evaluation indicated that most of these new compounds could potently inhibit FAK kinase, leading to the promising inhibitors against the proliferation of U-87MG, A-549, and MDA-MB-231 cancer cell lines. Among them, the optimized compound 18h potently inhibited the enzyme (IC₅₀ = 19.1 nM) and displayed stronger potency than TAE-226 in U-87MG, A-549 and MDA-MB-231 cells, with IC₅₀ values of 0.35, 0.24, and 0.34 μM, respectively. Compound 18h is a multi-target kinase inhibitor. Furthermore, compound 18h also exhibited relatively less cytotoxicity (IC₅₀ = 3.72 μM) toward a normal human cell line, HK2. According to the flow cytometry and wound healing assay results, compound 18h effectively induced apoptosis and G0/G1 phase arrest of MDA-MB-231 cells and suppressed the migration of U-87MG, A-549 and MDA-MB-231 cells. The docking study of compound 18h was performed to elucidate its possible binding modes and to provide a structural basis for the further structural guidance design of FAK inhibitors. Collectively, these data support the further development of compound 18h as a lead compound for FAK-targeted anticancer drug discovery.

黏着斑激酶（FAK）是负责各种肿瘤类型发展的细胞内非受体酪氨酸激酶。为了探索新的有效抑制剂，在基于结构的设计策略的基础上，设计并合成了两个系列的2,4-二取代-7 H-吡咯并[2,3- d ]嘧啶衍生物。生物学评估表明，这些新化合物中的大多数可以有效抑制FAK激酶，从而导致有希望的抑制剂对抗U-87MG，A-549和MDA-MB-231癌细胞的增殖。其中，优化的化合物18h 在U-87MG，A-549和MDA-MB-231细胞中均有效抑制酶（IC ₅₀ = 19.1 nM），并显示出比TAE-226更强的效力，IC ₅₀分别为0.35、0.24和0.34μM。化合物18h是多靶激酶抑制剂。此外，化合物18h 对正常人细胞株HK2的细胞毒性也较小（IC ₅₀ = 3.72μM）。根据流式细胞术和伤口愈合试验结果，化合物18h有效诱导MDA-MB-231细胞凋亡和G0 / G1期阻滞，并抑制U-87MG，A-549和MDA-MB-231细胞的迁移。进行化合物18h的对接研究以阐明其可能的结合方式，并为FAK抑制剂的进一步结构指导设计提供结构基础。这些数据共同支持化合物18h的进一步开发 作为FAK靶向抗癌药物发现的先导化合物。