Proteases play a key role in a variety of pathologies, including cancer, pancreatitis and thrombosis, which can be treated with low molecular inhibitors. Analyses of trypsin, thrombin and urokinase inhibition activity for selected extracts (BH4, diethyl ether; BH5, ethyl acetate, BH6, n-buthanolic) and flavonoids luteolin, cynaroside, (luteolin 7-O-glucoside), favanomarein from Bidens tripartia herb were determined. Quercetin was used as a positive control in tests on serine proteases (trypsin, thrombin and urokinase). BH5 showed a highly selective effect with a value of percentage inhibition activity (IA) = 91.68% for thrombin. Interesting results were observed in IA = 90.57% for BH6 for urokinase. The highest value of IA = 97.75% on plasmin was observed for BH4. Among the tested flavonoid compounds, the highest percentage inhibition activity with respect to the tested enzymes was shown by cynaroside indicating IA = 83.77-97.88%. The obtained results suggest that both extracts and the tested compounds are potential therapeutic resources for the development of serine proteases inhibitors.