New theobromine derivative as apoptotic anti-triple-negative breast cancer targeting EGFR protein: CADD story

A new EGFR inhibitor has been developed from theobromine (meta methoxy phenyl)acetamide derivative), T-1MMPA, exhibited the essential pharmacophoric characteristics needed to bind toEGFR's pocket. T-1-MMPA's anticancer potential was first estimated through various structure-based computational studies (DFT, docking, MD simulations over 200 ns, MM-GPSA, PLIP, ED, bi-dimensional and ADMET), which revealed that T-1-MMPA effectively bound to and inhibited the EGFR protein. The ADME and toxicity profiles of T-1-MMPA were also predicted computationally before the semi synthesis, and a high degree of drug-likeness was indicated. Then, T1-MMPA (2-(3,7-Dimethyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)-N-(3-methoxyphenyl)acetamide) was prepared to scrutinize the obtained in silico results. Subsequent in vitro studies showed that T-1-MMPA was effective against MDA-MB-231cell lines (triple-negative breast cancer), with an IC50 value of 1.42 & mu;M, compared to the reference drug (0.92 & mu;M) and exhibited a higher selectivity index of 1.9. Interestingly, T-1-MMPA also inhibited the growth of other three cancer cell lines (A549, CaCO-2, and HepG-2) with IC50 values of 1.57, 1.76, and 2.53 & mu;M, respectively. Additionally, T-1-MMPA effectively prevented the healing and migration abilities of the MDA-MB-231 cell lines, arrested the cell growth at the S phase and induced apoptosis, as confirmed by AO/ EB staining assay as well as the flow cytometry. Moreover, T-1-MMPA caused down-regulation of the BCL2 and MMP7 gene expression in the treated MDA-MB-231 cells. Finally, as the computational findings indicated T-1MMPA's hepatic safety, it was further corroborated through in vivo investigation.