Two inhibitors of urokinase were isolated from the extracts of the nuclei of peripheral pig leucocytes, by ion exchange chromatography and by gel filtration on Sephadex G-100 and G-150. Only the inhibitor with the higher molecular weight was isolated in the homogenous form, in a single polypeptide chain with molecular weight of 68,000, designated Inhibitor-3. This inhibitor has an isoelectric point from 4.4 to 4.5 and is stable in buffer solutions from pH 3 to 8, and belongs to the type of so-called fast reacting inhibitors.