Synthesis of new pyrazolo[3,4-d]pyrimidines as potential mutant EGFR/HER2 and Bcl2 inhibitors: anticancer evaluation, DFT, molecular docking and ADME studies

Abstract In this work, a novel 2-(6-methyl-4-oxo-4,5-dihydro-1 H -pyrazolo[3,4- d ]pyrimidin-3-yl)acetonitrile 4 was used as starting material to create a new series of pyrazolo[3,4-d]pyrimidines. DFT calculations and spectroscopic data (IR, 1 H NMR, 13 C NMR, and MS) were used to determine the structure of newly generated products. HepG2, MCF7, and HeLa were specific human cell lines used to evaluate these new compounds. When compared to erlotinib four specific compounds 6c , 8b , 10 and 13c showed strong cytotoxic effects. When erlotinib was used as the reference drug, both 8b and 10 exhibited notable effectiveness against EGFR T790M / HER2. A study of proliferation of breast cancer cell line showed that 8b increased in the percentage of DNA content in the G0-G1 phases, while 10 increased in the percentage of DNA content in the S phase. Furthermore, both 8b and 10 decreased in Bcl2 levels in breast cancer cells. Additionally, both compounds had favourable interaction with the EGFR tyrosine kinase domain (TKD) (PDB ID: 5JEB), with binding free energies (ΔG b ) close to -9.2 and − 7.5 kcal/mol, as revealed by molecular docking study. The molecular docking and ADME probe results were in accordance with the experimental data. In summary, the pyrazolo[3,4- d ]pyrimidines 8b and 10 displayed bioactivities on EGFR T790M / HER2, suggesting their potential possible preclinical candidates for future preclinical studies. Graphical Abstract