Focal adhesion kinase (FAK) is a ubiquitous intracellular non-receptor tyrosine kinase, which is involved in multiple cellular functions, including cell adhesion, migration, invasion, survival, and angiogenesis. In this study, a series of 7H-pyrrolo[2,3-d]pyrimidines were designed and synthesized according to the E-pharmacophores generated by docking a library of 667 fragments into the ATP pocket of the co-crystal complex of FAK and PF-562271 (PDB ID: 3BZ3). The 5-fluoro-7H-pyrrolo[2,3-d]pyrimidine derivatives demonstrated excellent activity against FAK and the cell lines SMMC7721 and YY8103. 2-((2-((3-(Acetamidomethyl)phenyl)amino)-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-N-methylbenzamide (16c) was selected for further bioactivity evaluations in vivo, including preliminary pharmacokinetic profiling in rats and toxicity assays in mice, and tumor growth inhibition studies in a xenograft tumor model. The results showed that 16c did not affect the body weight gain of the animals up to a dose of 200 mg/kg, and significantly inhibited tumor growth with a tumor growth inhibition rate of 78.6% compared with the negative control group. Furthermore, phosphoantibody array analyses of a sample of the tumor suggested that 16c inhibited the malignant proliferation of hepatocellular carcinoma (HCC) cells through decreasing the phosphorylation in the FAK cascade.

粘着斑激酶（FAK）是一种普遍存在的细胞内非受体酪氨酸激酶，参与多种细胞功能，包括细胞粘附、迁移、侵袭、存活和血管生成。在这项研究中，一系列的7 ħ吡咯并[2,3- d ]嘧啶根据由对接的667个片段的文库成FAK和的共晶体复合物的ATP袋产生的E-药效设计并合成PF-562271（PDB ID：3BZ3）。5-氟-7 H-吡咯并[2,3- d ]嘧啶衍生物对FAK和细胞系SMMC7721和YY8103表现出优异的活性。2-((2-((3-(乙酰氨基甲基)苯基)氨基)-5-氟-7 H-吡咯并[2,3 - d ]嘧啶-4-基)氨基)-选择 N-甲基苯甲酰胺 ( 16c ) 进行进一步的体内生物活性评估，包括大鼠的初步药代动力学分析和小鼠的毒性试验，以及异种移植肿瘤模型中的肿瘤生长抑制研究。结果表明，16c在高达200mg/kg的剂量下对动物的体重增加没有影响，与阴性对照组相比，对肿瘤生长有显着抑制作用，肿瘤生长抑制率为78.6%。此外，对肿瘤样本的磷酸抗体阵列分析表明，16c通过降低 FAK 级联中的磷酸化来抑制肝细胞癌 (HCC) 细胞的恶性增殖。