Based on structure-guided of thieno[2,3-d]pyrimidine skelecton functionalities and computer-aided drug development strategy, a series of novel 2,4-diamino-thieno[2,3-d]pyrimidine derivatives were designed and synthesized, and their structure confirmed by ¹H NMR, ¹³C NMR and HR-MS. All synthesized compounds were evaluated for antitumor activities against hepatic carcinoma (HepG2) cell lines in vitro, and results showed that all the target compounds exhibited potent antitumor activity. Preliminary comparison indicated that a more bulky substituent group at the C-4 position, and at the same time the introduction of 3-Cl on the phenyl ring with 4-methyl group on the C-2 position of thieno[2,3-d]pyrimidine have remarkable influence on activity. One of the expected target compound 5i showed the best inhibition activities against HepG2 with IC₅₀ value 0.17 μmol/L. To further investigate the structure-activity relationship, the docking experiment of 5i, 5j, 5o and Gefitinib on EGFR were performed. The docking results displayed that compound 5i was able to be tightly embedded in the active pocket of EGFR and the binding energy is most negative with EGFR, which demonstrating that compound 5i can be take a potential lead compound for further lucubrate.

基于噻吩并[2,3-d]嘧啶骨架功能的结构指导和计算机辅助药物开发策略，设计并合成了一系列新型2,4-二氨基-噻吩并[2,3-d]嘧啶衍生物，其结构经¹ H NMR、¹³ C NMR 和 HR-MS确证。所有合成的化合物均在体外评估了对肝癌（HepG2）细胞系的抗肿瘤活性，结果表明所有目标化合物均表现出有效的抗肿瘤活性。初步对比发现，噻吩并[2,3-d]的C-4位上有一个体积较大的取代基，同时在C-2位上有4-甲基的苯环上引入了3-Cl。 ]嘧啶对活性有显着影响。其中一种预期目标化合物5i对HepG2表现出最好的抑制活性，IC ₅₀值为0.17 μmol/L。为了进一步探讨构效关系，进行了5i、5j、5o和吉非替尼与EGFR的对接实验。对接结果显示，化合物5i能够紧密嵌入EGFR的活性口袋中，且与EGFR的结合能为最负，这表明化合物5i可以作为潜在的先导化合物进行进一步的研究。