The Prunus armeniaca L. (bitter apricot) is an apricot fruit tree categorized on the basis of the bitter taste of its seed kernel. In this study, the functional, medicinal, and therapeutic potential of bitter apricot seed kernel oil (BASKO) was evaluated. The qualitative screening of BASKO was performed using standard methodologies. The chemical profile of the oil was analyzed with the help of Fourier transform infrared (FTIR) and gas chromatography and mass spectrometry (GC-MS). Results revealed the presence of different phytochemical constituents comprising steroids, flavonoids, terpenoids, alkaloids, and cardiac glycosides. The antioxidant activity of the oil was determined by a 2,2,diphenyl-1picrylhydrazyl (DPPH) radical inhibition essay. Total phenolic and flavonoid contents were 10.6 ± 1.32 mg GAE/g and 4.75 ± 0.11 mg QE/g, respectively. DPPH inhibition of 89.5% was achieved at 1000 μg/mL of BASKO, with IC50 = 90.44 μg/mL (83.47-96.67 μg/mL with 95% CI). The antimicrobial potential of the BASKO revealed the inhibition of Escherichia coli (20.3 ± 2.08 mm), Salmonella typhi (19.3 ± 2.51 mm), Klebsiella pneumoniae (16.6 ± 1.52 mm), Pseudomonas aeruginosa (17 ± 2 mm), and Staphylococcus aureus (25 ± 1.01 mm). The minimum inhibitory concentration (MIC) value was 250 μL/mL for K. pneumoniae, S. typhi, P. aeruginosa, and S. aureus, whereas 62.5 μL/mL for E. coli. Moreover, BASKO showed antifungal potential against Trichophyton tonsurans (77.3 ± 2.08%), Epidermophyton floccosum (69.6 ± 3.51%), Aspergillus niger (74.3 ± 2.56%), Aspergillus flavus (90 ± 3%), and Mucor mucedo (78.3 ± 2.51%). Antileishmanial activity of oil was evaluated against Leishmania major by MTT assay, and an IC50 value of 89.75 μg/mL was observed. The study revealed that BASKO is a good source of biologically active compounds to be used as functional, therapeutical, and antimicrobial agents in food and pharmaceutical products.