Introduction: This study aimed to thoroughly profile the phytochemicals and pharmacological properties of Symphyotrichum squamatum's aerial parts to assess its potential as a valuable source of natural therapeutic agents. The research focused on isolating and identifying bioactive compounds and evaluating their antioxidant, cytotoxic, antidiabetic, and anti-inflammatory effects.Method: Chromatographic purification, LC/MS, total flavonoids, total phenolics, molecular docking and different pharmacological activity were used in this study to identify qualitatively and qualitatively the active compounds in extract and its activities.Results: Chromatographic purification of the petroleum ether and ethyl acetate fractions led to the isolation of four compounds: 3α- Friedelinol, Spinasterol, Dioctyl phthalate, and Kaempferol-3,7- diglucopyranoside. Additionally, the LC/MS profile of the hydro-alcoholic extract identified 35 metabolites, indicating a diverse chemical profile rich in fatty acids, phenolic propanoids, and terpenoids. Quantitative assays confirmed the extracts abundance in phenolics (65.9 ± 3.6 mg GAE/g extract) and flavonoids (25.8 ± 1.1 mg QE/g extract), correlating with notable in vitro antioxidant activity, as shown by low SC50 values of 77.00 μg/mL (DPPH) and 66.00 μg/mL (ABTS). The extract exhibited weak cytotoxicity against Hep-G2 and Panc-1 cell lines. Notably, both the extract and the isolated Kaempferol-3,7- diglucopyranoside demonstrated potent, dose-dependent inhibition of key carbohydrate-digesting enzymes, indicating antidiabetic activity. The flavonoid glycoside was particularly effective against α-amylase (IC50 = 24.29 μg/mL). The extract also showed promising anti-inflammatory activity via COX-1 inhibition (IC50 = 137.51 μg/mL). To explain these bioactivities, molecular docking studies were performed, revealing that the essential compounds, namely Kaempferol-3,7-diglucopyranoside and dicaffeoylquinic acids, form stable, high-affinity interactions with the reactive sites of α-amylase, α-glucosidase, and COX enzymes.Discussion: These findings collectively support S. squamatum as a promising candidate for further development in the management of diabetes and inflammation.