Therapeutic Potential of Three (3) Small Molecule Compounds and Mechanistic Insights into the Lead Compound for Cancer Treatment

Abstract

The need to discover novel anticancer drugs is imperative due to the high death rate associated with cancer. Conventional chemotherapy frequently lacks selectivity and induces significant adverse reactions. This study sought to identify specific small-molecule compounds that can selectively interact with cancer cells while causing minimum harm to normal cells. The cytotoxic effects of three small molecule compounds (compounds 1, 2, 3) were evaluated on different cancer cell lines. The use of computational tools, such as molecular docking and dynamics simulations, facilitated the prediction of pharmacokinetics properties and the identification of molecular targets. The effects of the most potent compound on cellular processes such as apoptosis, gene expression, cancer stemness, and migration in breast cancer cells were investigated using a combination of flow cytometry, real-time PCR, adhesion and wound healing assays. A set of 36 analogues was developed using the structure of the hit compound as a basis. Eight of these analogues were tested for their cytotoxic effects, pharmacokinetic features, and drug-likeness. Compound 2, the identified hit compound, demonstrated significant cytotoxicity against breast cancer cell lines, surpassing doxorubicin (DOX) in selectivity. The molecular targeting analysis revealed substantial interactions with cancer-related pathways, specifically EGFR, AKT1, and VEGFR2. EGFR was subsequently identified as the most likely target, with compound 2 binding to its ATP binding pocket and allosteric site. Compound 2 was shown to have the capacity to trigger apoptosis, hinder the migration of cancer cells, and modify gene expression associated with cancer metastasis, as demonstrated by functional experiments. It also modulated inflammation and immune responses differentially from DOX. Although compound 2 showed promising efficacy, its analogues did not exhibit increased activity, suggesting selectivity in its mechanism of action. This study demonstrates the therapeutic potential of Compound 2 as an anticancer agent by targeting EGFR and possibly other pathways. The results confirm the compound's ability to regulate critical cancer related processes and pathways, providing a basis for further development into a cancer treatment that can be used in clinical settings. The findings support further development and optimisation of Compound 2 and its analogues for further investigations.