Design, synthesis, and biological evaluation of novel 4-(4-ethoxyphenyl)-6-(substituted-phenyl)pyrimidin-2-amine/thiol/ hydroxy derivatives as EGFRWT and EGFR T790M inhibitors targeting NSCLC: In-vitro and in-silico exploration

The novel series of 4-(4-ethoxyphenyl)-6-(substituted-phenyl)pyrimidin-2-amine/thiol/hydroxy were designed, synthesized screened for anticancer activity as EGFRWT and EGFR T790M inhibitors. Newly, developed compounds ( RT-1 - RT-12) were screened against cytotoxicity assay using brine shrimp cytotoxicity methods. The results described that all the derivatives were better than doxorubicin (LD50: 120.02 f 0.85 mu g/mL) with an LD50 value range from 65.03 f 01.35-80.54 f 02.25 mu g/mL. Further, the series of derivatives were examined against three cell lines (A549, NCI-H1975, and NCI-H522) of non-small cell cancer (NSCLC). Compounds, RT-8 ( IC 50 : 1.56 f 00.56 and 0.95 f 02.80 mu M), RT-10 ( IC 50 : 1.59 f 00.56 and 0.99 f 01.06 mu M), and RT-11 ( IC 50 : 1.55 f 01.25 and 0.91 f 01.25 mu M) were the most potent against the A549, and NCI-H1975 cancer cell lines. Whereas, only two compounds RT-8 and RT-9 were most potent against the NCI-H522 cell line with IC50 values of 1.01 f 00.69 and 0. 99 f 00.85 mu M corresponds to the standard, afatinib (IC50: 1.03 f 01.25 mu M) respectively. Compounds RT-8 and RT-11 were evaluated for enzyme inhibition activity against EGFRWT and EGFRT790M. The results revealed that compounds RT-8 (IC50 = 0.020 f 013.74 and 0.035 f 02.72 mu M) and RT-11 (IC50: 0.018 f 017.11 and 0.020 f 01.02 mu M) exhibited potent inhibitory activities towards EGFRWT and EGFRT790M. The ADMET parameters were forecasted with online tools SwissADME and Protox. Most of the compounds were in the predictable range and none of the compounds violated the Lipinski and Veber rules. Finally, molecular docking analysis also supports an in-vitro study and describes the good binding pattern of RT-8 and RT-11 against, EGFRWT and EGFRT790M. The molecular simulation study also confirmed that RT-11 was stable with EGFRT790M. Furthermore, this work serves as a foundation for future studies on EGFR inhibitors. These encouraging results present the pharmacophoric features of 4-(4-ethoxyphenyl)-6-(substituted-phenyl)pyrimidin-2-amine/thiol/hydroxy as an interesting lead for the further development of new EGFRWT and EGFR T790M drug candidate.