A series of 3-N-aryl substituted-2-heteroarylchromones was efficiently synthesized via Pd-mediated oxidative coupling under the influence of hetero-atoms neighboring group participation. Synthesized molecules were evaluated against human microtubule affinity regulating kinase 4 (MARK4) enzyme inhibitors, a recently identified anti-cancer drug target. Among 21 synthesized molecules, compounds 2, 3, and 14 exhibited excellent in vitro inhibitory effect against MARK4 with IC value (50% of ATPase activity) at 2.12 ± 0.22 μM, 1.98 ± 0.34 μM and 5.56 ± 0.42 μM respectively. The fluorescence binding and dot blot assay of these compounds were found in μM range, indicating a better binding affinity. In vitro study of these compounds against cancerous cells (MCF-7 and HepG2) inhibited the cell viability, induced apoptosis and lowered the tau-phosphorylation. Cell viability studies of compounds 2, 3, and 14 showed inhibition of cancerous cells growth with IC values of 3.22 ± 0.42, 4.32 ± 0.23 μM and 16.22 ± 1.33 μM for human breast cancer cells (MCF-7) and 6.45 ± 1.12, 5.22 ± 0.72 μM and 19.12 ± 1.43 μM for human liver carcinoma cells (HepG2) respectively. ROS quantification of these compounds showed oxidative stress to cancerous cells and molecular docking study showed hydrogen bonding, charge or polar and van der Waals interactions with the active site residues of MARK4. Owning to high binding fit nicely in the active site, offering the possibilities to be used as novel therapeutic molecules in the drug discovery against MARK4-related diseases.