Fifty-six methanol extracts obtained from the barks, flowers, leaves and stems of 30 Slovak trees, bushes and herbs used in the traditional medicine of the Small Carpathians, Slovakia, have been screened for antiprotease (trypsin, thrombin and urokinase) activity using chromogenic bioassay. In this study, 14 extracts showed the strong inhibition activity to protease trypsin with IC50 values below 10 microg/mL. The highest inhibition activities were observed for methanol extracts of Acer platanoides IC50 = 1.8 microg/mL, Rhus typhina IC50 = 1.2 microg/mL and Tamarix gallica IC50 = 1.7 microg/mL. However, the results of extracts tested on thrombin were generally different from those observed for trypsin. The most marked inhibition activity to thrombin were estimated for extracts of Castanea sativa IC50 = 73.2 microg/mL, Larix decidua IC50 = 96.9 microg/mL and Rhus typhina IC50 = 20.5 microg/mL. In addition, Acer platanoides and Rhus typhina were the only extracts which showed inhibition activity to urokinase with IC50 = 171.1 microg/mL and IC50 = 38.3 microg/mL, respectively. In addition, Rhus typhina showed the broadest spectrum of inhibition activity to all tested serine proteases and seems to be a prospective new source of natural products as inhibitors of serine proteases.