Herbal preparations have been used since ancient times for the treatment of several diseases and constitute a major share of all the officially recognized systems of health in India. The aim of the present study was to assess the in vitro thrombolytic activity of leaf extracts from Leucas aspera, Cynodon dactylon and Murraya koenigii in combination with clot buster enzymes (i.e., Nattokinase-NK and Staphylokinase-SaK). Methanolic leaf extract of L. aspera, C. dactylon and aqueous leaf extract of M. koenigii were prepared and assessed for their thrombolytic activity by clot lysis assay. The average clot lysis activity of L. aspera, C. dactylon and M. koenigii were found to be 45.32±0.53%, 38.56±0.42% and 30.32±0.25%. Combination of clot buster enzymes and leaf extracts showed an maximum clot lysis activity at the proportional ratio 1:1 of NK:L. aspera (47.61±0.35%), NK:C. dactylon (55.88±0.52%), SaK:L. aspera (59.04±0.54%) and SaK:C. dactylon (50.75±0.56%). GC–MS analysis of L. aspera, C. dactylon and M. koenigii extracts revealed the presence of thrombolytic compounds. This is the first report on combined thrombolytic activity of clot buster enzymes and selected herbal extracts. Hence this study demonstrates the combinational properties of the herbal extracts and clot buster enzymes which could be one of the best sources of herbal drugs for thrombotic disorders with minimal side effects. Advances in this research could lead to the development of new combination of thrombolytics. © 2016 Elsevier Ltd