Design, Synthesis, Biological Evaluation, and In Silico Studies of Novel Furo[2,3‐ <i>d</i> ]pyrimidine Derivatives as EGFR Inhibitors With Potential Antitumor Activity

In the pursuit of developing targeted therapies, novel furo[2,3-d]pyrimidine compounds were synthesized, incorporating the characteristic structural features of EGFR tyrosine kinase inhibitors. These compounds were screened for antiproliferative effects across 60 human cancer cell lines. Compound 3f was the most effective. Further evaluation at five concentrations to determine its GI₅₀, TGI, and LC₅₀. In vitro studies were conducted to evaluate its impact on EGFR signaling, cell cycle progression, apoptosis induction, and caspase-3 activation in T-47D cells. Compound 3f showed greater selectivity toward cancerous T-47D cells over normal breast cells (MCF10a). Notably, compound 3f displayed potent EGFR inhibition at submicromolar levels, with an IC₅₀ of 0.121 ± 0.004 μM, which was comparable to the reference inhibitor, erlotinib. Further investigations revealed that compound 3f caused cell cycle arrest at the G2/M phase in T-47D cells. Apoptosis analysis confirmed that compound 3f induced cell death through programmed cell death, with a nearly 19-fold increase in total apoptosis. This apoptotic mechanism was validated by a substantial rise in active caspase-3 levels. Molecular docking studies showed favorable binding of compound 3f within the EGFR active site. Molecular dynamics simulations further confirmed the stability of compound 3f within the active site over a 100-ns simulation period, supporting its sustained interaction with key residues. Based on predictions of toxicity, compound 3f was predicted to possess a favorable safety profile across 30 potential toxicities. Encouraged by its strong anticancer efficacy and safety profile, compound 3f represents a compelling lead candidate for further development as a targeted treatment for breast cancer.