The primary goal of this study is to synthesize and characterize N-substituted Acetamido derivatives of acridone, where the acetamido moiety has been considered a linker which is crucial for several biological activities, including anti-cancer activity. In this context, the anti-proliferative activity of synthesized derivatives was evaluated against human breast (MCF-7, MDA-MB-231), lung (A-549), and skin (A-431) cancer cell lines. Results revealed that compounds 8 h, 8i, 9 h, and 9i showed the most potent activity against MCF-7 cell lines with IC50 values of 13.96 µM, 8.25 µM, 9.45 µM, and 6.76 µM, respectively. In addition, all these compounds were found to be non-toxic against normal cells (NIH/3T3). Further, AKT kinase inhibition assay results showed that compounds 8i and 9i have the efficacy to inhibit the AKT kinase with potential anti-cancer activity. The cell cycle analysis revealed that compounds 8i and 9i could arrest the G0/G1 phase of the cell cycle and absorption titration with CT-DNA identified that these molecules could interact with DNA. In order to understand the drug-likeness properties, all the compounds were evaluated by various in silico screening, and these compounds exhibited optimal physicochemical features as excellent lead molecules. Finally, in vitro results were validated using a molecular docking study, which revealed binding interactions in the active site of AKT.