This study presents a comprehensive analysis of purine analogues, focusing on their biological evaluations, including antioxidant, cytotoxic activity, and enzyme inhibition, alongside in-depth computational assessments using density functional theory (DFT) and molecular docking. A series of purine analogues were synthesized, characterized, and subjected to antioxidant assays to determine their radical scavenging capabilities, and cytotoxicity was evaluated against various cancer cell lines (A549 and DLD-1) to assess their potential therapeutic efficacy. The experimental findings indicated that certain purine derivatives exhibited significant antioxidant properties and cytotoxic effects, highlighting their potential as bioactive compounds. Furthermore, the synthesized purine analogues showed α-glucosidase enzyme inhibitory activity at all concentrations investigated except molecule 2a. The geometry of compounds was optimized under the DFT/B3LYP/6–311++G (d,p) level of calculation in order to investigate the electronic behaviors. The MEP surface and HOMO-LUMO analysis of compounds were studied to predict the reactive sites for electrophilic and nucleophilic attacks and biological activity and stability. Molecular docking simulations against two distinct protein receptors (PDB ID: 3KJF and 3WZE) to explore their impact on cancer cells, considering the promising cytotoxic activity of compounds.