Anti-aggregation activity of crude water extract of Galega officinalis L. fractionated on Sephadex G-25 and Sepharose 4B.

The present study describes a method for preparation of biologically active fraction from crude water extract of Galega officinalis L. by gel filtration on Sephadex G-25 and Sepharose 4B. In an in vitro experiment (at a dose of 12.0 +/- 0.45 micrograms/ml) fractionated extracts inhibited adenosine diphosphate (ADF) induced platelet aggregation by 50%. Inhibitory effects on collagen (0.18 mg/ml) and thrombin (0.7 U/ml) induced platelet aggregation were observed at doses of 0.18 +/- 0.65 microgram/ml and 20 +/- 0.82 micrograms/ml, respectively. The optimum activity was observed at a temperature of 30-42 degrees C. It was found that the fraction contained 15.23% protein. As shown by amino acid analysis several amino acids (alanine, glycine, valine, lysine, asparagine, arginine and serine) accounted for 50% of its protein content. These amino acids formed tri- and tetrapeptides (RGD, RGDS, KRDS, RGDS), which inhibited platelet aggregation. The RGD and AGVD fibrinogen amino acid sequences responsible for the recognition and binding to the glycoprotein IIb/IIIa receptors consisted of the same amino acids.