Isolation, chemophenetic profiling, antibacterial activity, and in silico pharmacokinetic-toxicological evaluation of secondary metabolites from Viola canescens Wall. ex Roxb.

Viola canescens Wall. ex Roxb., a medicinal species native to the Western Himalayas, is traditionally used to treat inflammatory and infectious disorders. In this study, eight secondary metabolites kaempferol, luteolin, quercetin, 4-hydroxycoumarin, β -sitosterol, palmitic acid, myo-inositol, and o -coumaric acid were isolated from flower extracts and structurally characterized using UPLC, HRMS, and NMR spectroscopy. Antibacterial activity was evaluated against six Gram-positive and Gram-negative bacterial strains using agar well diffusion and broth microdilution assays. Luteolin and o -coumaric acid exhibited the strongest activity, with inhibition zones up to 28.33 ± 1.25 mm and MIC values as low as 1.25 mg/mL. Molecular docking against COX-2 (PDB ID: 1CX2 ) revealed strong binding affinity of quercetin (-10.438 kcal/mol), comparable to the reference inhibitor SC-558. SwissADME analysis indicated favorable gastrointestinal absorption and drug-likeness for polyphenolic metabolites, while ProTox-III predictions classified most compounds in low toxicity categories (GHS IV–VI). These findings support the chemophenetic and pharmacological relevance of V. canescens metabolites. The current computational findings suggested anti-inflammatory potential of V. canescens but detailed validation of its claims through in-vivo studies are still required to increase the utilization of V. canescens.