Studies On FXa Inhibitors Of Vaccariae Semen, Persicae Semen And Curcumae Radix

Thrombotic disease has seriously endangered human health, but the traditional anticoagulants have many restrictions and potentially fatal risks, such as narrow therapeutic window, easy to cause hemorrhagic disease, need clinical monitoring and so on. So it has been replaced by new anticoagulants owed better safety and effects. Modern research shows the inhibition of factor Xa (FXa) activities can effectively anticoagulat without affecting the underlying hemostatic, so FXa inhibitors have become the ideal anticoagulants. The exploration of new type active compounds to inhibit FXa is important.Traditional Chinese medicine has been used for a long history for the treatment of arterial and venous thrombosis, and some pharmacological research also showed that parts of Traditional Chinese medicine have certain anticoagulant activities. In the previous study, we screened the potential inhibitory activity of 26 species of those plants which were used as traditional Chinese medicine for the treatment of thrombosis. And the result showed that the ethanol extracts of Vaccariar Semen, Persicae Semen and Curcumae Radix possessed inhibitory activities against the FXa. On the basis of prophase research, this research isolated and purified bioactive ingredients form the traditional Chinese medicine to obtain a new research direction for future development of anticoagulants owed independent intellectual property rights. The main research contents:1. Studies on FXa inhibitors of Vaccariar SemenOn the basis of spectroscopic methods, thirteen compounds were isolated and identified, including a-spinasterol, N-(3-methoxy-4-hydroxyphenethyl)-tetracosanamide, stigmast-1, 5-dien-3-ol, N-(3,4-dihydroxyphenethyl)-tetracosanamide, 16-hydroxygypsogenic acid, gypsogenic acid, protocatechuic acid, stigmast-7, 22-dien-3-one, Segetalins A, Segetalins B, Adenosine, Vitexin and pinnatifin E. And all compounds were tested for their FXa inhibition activities and showed weak inhibition activity to FXa with the IC50 of 279.55μmol/L,286.98 μmol/L,307.54μmol/L, 210.57μmol/L,222.67μmol/L,165.23μmol/L,293.67μmol/L,654.65 μmol/L,198.18μmol/L,249.30μmol/L,380.71μmol/L,221.83μmol/L, and 278.17μmol/L (all P<0.01), respectively.2. Studies on FXa inhibitors of Persicae SemenOn the basis of spectroscopic methods, ten compounds were isolated and identified, including 9-Octadecenoic acid-2,3-dihydroxypropyl ester, daucosterol, 2-hydroxy-2-phenylacetamide, benzoic acid, 2-hydroxy-9-octadecenoic acid, n-heptadecanoic acid, methyl elaidate, p-hydroxybenzoic acid, hexadecanoic acid and amygdalin. And all compounds were tested for their FXa inhibition activities and showed weak inhibition activity to FXa with the IC50 of 265.80μmol/L,187.25μmol/L,565.46μmol/L,850.54μmol/L,454.06 μmol/L,303.52μmol/L,440.56μmol/L,691.75μmol/L,373.58μmol/L and 220.12μmol/L (all P<0.01), respectively.3. Studies on FXa inhibitors of Curcumae RadixOn the basis of spectroscopic methods, ten compounds were isolated and identified, including gweicurculactone, a-epoxide, hexadecanoic acid, trilinolein, di-n-butyl phthalate, 1-docosanol, curcumenol, β-sitosterol, 5-hydroxymethyl-furaldehyde, 5,5-oxydimethylene-bis-(2-furfural). And all compounds were tested for their FXa inhibition activities and showed weak inhibition activity to FXa with the IC50 of 287.31μmol/L,233.78 μmol/L,303.55μmol/L,376.83μmol/L,402.31μmol/L,301.20μmol/L,202.39μmol/L,255.90 μmol/L,173.61μmol/L and 166.88μmol/L (all P<0.01), respectively.4. Synergistic effect of a factor Xa inhibitor, Glycyrretinic acid, and antiplatelet agents on whole blood coagulation and arterial thrombosis in ratsIn vitro, the blood was sampled by the method of abdominal aorta in the rats and the plasma was prepared to determine activeated partial thromboplastin (APTT), thrombin time (TT). And synergistic action on anticoagulation of glycyrretinic acid and antiplatelet agents were evaluated on the methods of Berenbaum. The results showed that a factor Xa inhibitor, glycyrretinic acid, pius tetramethylpyrazine (TMP) markedly prolonged APTT and TT time. In vivo, the coagulation time (CT) was measured using the capillary methods, and the bleeding time was measured using tail-cutting methods. The bleeding time and coagulation time was prolonged after the treatment by synergistic action of glycyrretinic acid and tetramethylpyrazine.This paper studied on the active compounds with inhibition to FXa from Vaccariar Semen, Persicae Semen and Curcumae Radix. It resulted that gypsogenic acid, Segetalins A, daucosterol, 5-hydroxymethyl-furaldehyde and 5,5- oxydimethylene-bis-(2-furfural) both had some inhibition to FXa. And it also studied on the synergistic effect of Glycyrretinic acid and antiplatelet agents on whole blood coagulation and arterial thrombosis in rats. It resulted that a factor Xa inhibitor, glycyrretinic acid, pius tetramethylpyrazine (TMP) markedly prolonged APTT/TT time and the coagulation time without markedly bleeding. Thought these compounds have weak activities compared with listed drugs, their different structure types provide a new directive for future development of FXa inhibitors.