Department of Medicinal Chemistry, School of Pharmacy, Fudan University, Shanghai, 201203, China.
Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, 200240, China.
Eur J Med Chem. 2021 Nov 5;223:113670. doi: 10.1016/j.ejmech.2021.113670. Epub 2021 Jun 25.
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.
黏着斑激酶（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，包括细胞黏附、迁移、侵袭、存活和血管生成。在这项研究中，根据将 667 个片段库对接入 FAK 和 PF-562271（PDB ID：3BZ3）共晶复合物的 ATP 口袋中产生的 E-药效团，设计并合成了一系列 7H-吡咯并[2,3-d]嘧啶。5-氟-7H-吡咯并[2,3-d]嘧啶衍生物对 FAK 和 SMMC7721 和 YY8103 细胞系表现出优异的活性。2-((2-((3-(乙酰氨甲基)苯基)氨基)-5-氟-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)-N-甲基苯甲酰胺（16c）被选为进一步的体内生物活性评估，包括在大鼠中的初步药代动力学特征研究和在小鼠中的毒性试验，以及在异种移植肿瘤模型中的肿瘤生长抑制研究。结果表明，16c 高达 200mg/kg 的剂量不会影响动物的体重增加，并与阴性对照组相比，显著抑制肿瘤生长，肿瘤生长抑制率为 78.6%。此外，肿瘤样本的磷酸化抗体阵列分析表明，16c 通过降低 FAK 级联中的磷酸化来抑制肝癌（HCC）细胞的恶性增殖。
