Colorectal cancer (CRC) is a highly aggressive malignancy characterized by rapid progression, a strong tendency to metastasize, and poor clinical outcomes. The synthetic compound IK-2550, chemically identified as N-(3-(imidazo[1,2-a]pyridin-2-yl)phenyl)-2-methylbenzamide (molecular weight: 327.39 g/mol), is a novel small molecule whose anticancer mechanisms remain largely unexplored in HCT116 colorectal cancer cells. This study evaluates the anticancer activity of IK-2550 in HCT116 cells and investigates its underlying signaling mechanisms. Using MTS assays, flow cytometry, and Western blot analysis, the effects of IK-2550 on cell growth, proliferation, and apoptosis were assessed in vitro. Treatment with IK-2550 significantly reduced cell viability, increased the Bax/Bcl-2 ratio, promoted cytochrome C release, and activated the caspase cascade, leading to the cleavage of poly-(ADP-ribose) polymerase (PARP). Additionally, IK-2550 downregulated murine double minute-2 (Mdm2) expression while enhancing p53 levels. One of the most critical signaling axes in CRC, the PI3K/Akt/mTOR pathway, is frequently dysregulated, driving tumor initiation, progression, metastasis, and drug resistance. IK-2550 was found to inhibit this pathway by decreasing the phosphorylation of PI3K, Akt, and mTOR, thereby triggering apoptosis in HCT116 cells. In addition, the generation of ROS was increased during IK-2550-induced apoptosis. Collectively, these findings highlight IK-2550 as a potential therapeutic candidate for colorectal cancer through its ability to suppress the PI3K/Akt/mTOR pathway and induce apoptosis.