Background: Diabetes prevalence is progressively rising everywhere, particularly in developing countries. Lichens have evolved into a rich source of innovative bioactive chemicals with their anti-oxidant capabilities, widening the scope of well-documented and effective diabetes treatment. Objective: The main objectives of this study were to perform in vitro alpha-amylase, alphaglucosidase inhibition analysis, and anti-oxidant of lichen Physica aipolia. Method: Folin Ciocalteu’s reagent was used to determine the total phenolic content for biological activities. The Aluminum trichloride method was used to determine total flavonoid content, and the free radical assay method was used to determine the antioxidant activity of P. aipolia using 2,2- diphenyl-1-picrylhydrazyl (DPPH). Moreover, substrate 2-Chloro-4-nitrophenyl-α-D-maltotrioside (CNPG3) and substrate-nitrophenyl-α-D-glucopyranose (p-NPG) were used for the determination of alpha-amylase and alpha-glucosidase inhibition activities respectively, and molecular docking was performed by Auto Dock Vina. Results: Total phenolic and flavonoid contents present in P. aipolia were 37.41 ± 2.87 mg GAE/g and 5.53 ± 0.95 mg QE/g, respectively. For anti-oxidant inhibition activity, crude extract of methanol showed an IC50 value of 15.324 ± 0.80 μg/mL. Furthermore, a methanolic crude extract showed significant inhibition activities against alpha-amylase (IC50 = 178.50 ± 1.10 μg/mL) and alphaglucosidase (IC50 = 76.10 ± 0.91 μg/mL). During in silico analysis, zeorin showed an effective binding affinity with -7.9 kcal/mol, and Atranorin showed -7.4 kcal/mol in the orthosteric site of the protein. Conclusion: Physica aipolia exhibits the potential to impact biological functions, demonstrating antidiabetic properties, as confirmed by molecular docking analysis in silico.