Design, synthesis and anticancer evaluation of 1H-pyrazolo[3,4-d]pyrimidine derivatives as potent EGFRWT and EGFRT790M inhibitors and apoptosis inducers

Abstract In our attempt to develop effective EGFR-TKIs, two series of 1 H -pyrazolo[3,4- d ]pyrimidine derivatives were designed and synthesized. All the newly synthesized compounds were evaluated in vitro for their inhibitory activities against EGFR WT . Compounds 15 b , 15 j , and 18 d potently inhibited EGFR WT at sub-micro molar IC 50 values comparable to that of erlotinib. Moreover, thirteen compounds that showed promising IC 50 values against EGFR WT were tested in vitro for their inhibitory activities against mutant EGFR T790M . Compounds 17 d and 17 f exhibited potent inhibitory activities towards EGFR T790M comparable to osimertinib. Compounds that showed promising IC 50 values against EGFR WT were further tested for their anti-proliferative activities against three cancer cell lines bearing EGFR WT (MCF-7, HepG2, A549), and two cancer cell lines bearing EGFR T790M (H1975 and HCC827). Compounds 15 g , 15 j , 15 n , 18 d and 18 e were the most potent anticancer agents against the EGFR WT containing cells, while compounds 15 e , 17 d and 17 f showed promising anti-proliferative activities against EGFR T790M containing cells. Furthermore, the most active compound 18 d was selected for further studies regarding to its effects on cell cycle progression and induction of apoptosis in the HepG2 cell line. The results indicated that this compound is good apoptotic agent and arrests G 0 /G 1 and G 2 /M phases of cell cycle. Finally, molecular docking studies were performed to investigate binding pattern of the synthesized compounds with the prospective targets, EGFR WT (PDB: 4HJO) and EGFR T790M (PDB: 3W2O).