Oral squamous cell carcinoma (OSCC) represents the most prevalent sub-type of oral malignancy, characterized by high morbidity and mortality despite ongoing advances in treatment strategies. BaLac, a traditional Ayurvedic formulation derived from Berberis aristata DC, has been used for managing oral ailments like neoplastic lesions. This study evaluated the anticancer potential of BaLac against OSCC and identified lead bioactive candidates with promising pharmacological characteristics through an inte-grative in vitro and in silico approach. The cytotoxic activity of BaLac was determined in KB cells via MTT assay, while apoptosis induction was validated through acridine orange-ethidium bromide dual staining. Phytochemical constituents identified from BaLac and Berberis aristata through UHPLC-MS were subsequently subjected to molecular docking against phosphoinositide 3-kinase (PI3K), a pivotal signaling molecule in OSCC pathogenesis. Binding stability and pharmacokinetic properties were further analyzed using molecular dynamics simulation, Lipinski’s rule, and ADMET profiling. BaLac exhibited dose-dependent cytotoxicity with an IC₅₀ of 573.01 μg/mL and induced apoptosis in KB cells. Virtual screening identified several phytocompounds with strong PI3K-binding affinities, exceeding that of the reference inhibitor buparlisib. Among the screened candidates, homocodeine (unique to BaLac) and (15Z)-9,12,13-trihydroxy-15-octadecenoic acid met drug-likeness and safety criteria, indicating their potential as orally active agents. Molecular dynamics analysis confirmed stable and sustained binding of these compounds within the PI3K catalytic pocket. Overall, BaLac demonstrated significant anticancer properties against OSCC, and its phytoconstituents showed predicted interactions with PI3K based on in silico analyses, supporting further experimental validation, toward its development as a therapeutic candidate for OSCC.