Imidazo[1,2-a]Quinoxaline-2-Carbonitrile Derivative (RA-22) Inhibits Self-Renewal and Growth of Cancer Stem and Cancer Cells via Downregulating AKT Pathway

Abstract

EGFR activation in colorectal and breast cancer stimulates downstream pathways like Ras/Raf/MEK/ERK and PI3 K/Akt, fostering cell proliferation, invasion, metastasis, and therapy resistance, underscoring its significance as a therapeutic target in both cancers. In the present work, we rationally designed (E)-4-methyl-1-((3-oxo-1-phenylbutyl)amino)-4-styryl-4,5-dihydroimidazo[1,2-a]quinoxaline-2-carbonitrile (RA-22) as EGFR inhibitor. Our research investigates the role of RA-22 as a target molecule for EGFR, exploring its anticancer potential and mechanism of action across breast cancer and colorectal cancer cell lines. The in-vitro studies showed its cytotoxic response towards MDA-MB-231 and HCT-116 and its inhibitory effect on cancer stem cells in mammosphere/spheroid culture. The compound downregulates the oncogenic signalling proteins like STAT-3, AKT, PAN-AKT, and ERK and also reduces the expression of the anti-apoptotic protein Bcl-2 and increases the apoptotic proteins like Cleaved-PARP, Cleaved-Caspase-3, and Cleaved-Caspase-9.