CN108785299A

CN108785299A - Glabridin is preparing the application in treating thrombotic diseases drug

Info

Publication number: CN108785299A
Application number: CN201710292058.1A
Authority: CN
Inventors: 萧伟; 陶晓倩; 苏真真; 王俨如; 丁岗; 王振中
Original assignee: Jiangsu Kanion Pharmaceutical Co Ltd
Current assignee: Jiangsu Kanion Pharmaceutical Co Ltd
Priority date: 2017-04-28
Filing date: 2017-04-28
Publication date: 2018-11-13

Abstract

The invention discloses glabridins to prepare the application in treating or preventing thrombotic diseases drug, belongs to pharmaceutical technology field.Experiment shows that glabridin has apparent inhibiting effect to platelet activating factor receptor；FeCl can be significantly reduced in vivo₃Solution is applied with p- selectins in the thrombus model rat plasma caused by arteria carotis, II b/ of vWF, GP III a and TXB2 levels；Glabridin intravenous injection can significantly inhibit the whole blood PAF concentration that Arterial thrombosis is induced and increase, and can significantly reduce IP in blood platelet₃Content and blood platelet IP₃The expression of receptor shows compounds on platelet PAF-IP₃‑IP₃R‑[Ca²⁺] i signal transduction pathway inhibiting effect, it is one of the mechanism of glabridin D-dimer and aggregation.Glabridin can have the function for the treatment of thrombotic diseases and value with multiple target point, the inhibition thrombosis of multi-path.

Description

Glabridin is preparing the application in treating thrombotic diseases drug

Technical field

The invention belongs to pharmaceutical technology fields, are related to glabridin in preparing treatment or prevention thrombotic diseases drug Using.

Background technology

Thrombotic disease is the current disease for seriously endangering middle-aged and the old's health, and incidence is high, lethality of disabling It is high.Thrombosis is the complex process of multifactor participation, progressive development, and blood because of the activation of blood platelet and coagulates under flow regime Blood factor is activated and the false setting that occurs.Thrombosis plays a driving role in the generation and development of cardiovascular and cerebrovascular disease, And one of the main reason for causing death and disabling.The condition of thrombosis includes that blood vessel internal membrane damage, blood flow state change Become and blood coagulability increases.Therefore, inhibit platelet function and prevent blood clotting can antithrombotic.Clinically apply Antiplatelet and antithrombotic drug it is more, such as blood platelet cox-2 inhibitors aspirin, blood platelet cyclic nucleotide Inhibitor Dipyridamole, adenosine diphosphate (ADP) (ADP) receptor antagonist clopidogrel, the heparin of antithrombase Ш (AT Ш), dimension life Plain K antagonists warfarin etc., the common feature of these drugs are single medicine, single target spot and irreversible inhibition, lead to adverse reaction It happens occasionally.Therefore, multiple target point, multi-path and significant in efficacy, safe antithrombotic reagent is found to be of great significance.Closely The research of Nian Lai, Chinese medicine treatment thrombotic diseases gradually cause the concern of people, and since many traditional Chinese medicinal components belong to natural Drug is extracted, has many advantages, such as Small side effects.

Radix Glycyrrhizae Glycyrrhizae Radix et Rhizoma. are legume Glycyrrhiza Uralensis (Glycyrrhiza Uralensis Fisch.), swollen fruit Radix (Glycyrrhiza inflata Bat.) or glycyrrhiza glabra (Glycyrrhiza Glabra L.) drying root and rhizome, be a kind of common Chinese medicine.The flavones contained in studies have shown that Radix Glycyrrhizae in recent years Constituents have the effects that stronger anti-oxidant, antiatherosclerosis and certain reducing blood lipid, blood pressure lowering.

Glabridin (glabridin, C₂₀H₂₀O₄, No. CAS：59870-68-7) it is specific flavonoids in glycyrrhiza glabra Ingredient, and proportion is larger in glycyrrhiza glabra flavones ingredient, about 11%, structural formula is shown in formula I：

Before glabridin also shows that good research in the research that Prevention of cardiovascular disease drug and cosmetics are developed Scape.Modern pharmacological studies have shown that glabridin have adjust immune, reducing blood lipid, hypoglycemic, oestrogen-like hormone, inhibit tyrosinase, The effects that antibacterial, anti-inflammatory, anti-oxidant, anti-osteoporosis.

But have no that glabridin is preparing the report treated and applied in thrombotic diseases drug at present.

Invention content

The purpose of the present invention is to provide glabridins to prepare the application in treating or preventing thrombotic diseases drug.It is real It tests and shows glabridin to having apparent inhibiting effect to platelet activating factor receptor (PTAFR)；It can significantly drop in vivo Low FeCl₃Solution is applied with p- selectins, II b/ of vWF, GP, III a and TXB2 water in the thrombus model rat plasma caused by arteria carotis It is flat；Glabridin intravenous injection can significantly inhibit the whole blood PAF concentration that Arterial thrombosis is induced and increase, and can significantly drop IP in low platelet₃Content and blood platelet IP₃The expression of receptor shows compounds on platelet PAF-IP₃-IP₃R-[Ca²⁺]i Signal transduction pathway inhibiting effect is one of the mechanism of glabridin D-dimer and aggregation.Show that glabridin can be with The inhibition thrombosis of multiple target point, multi-path has the function for the treatment of thrombotic diseases and value.

The purpose of the present invention is what is be achieved through the following technical solutions：

Application of the glabridin in preparing the drug for treating or preventing thrombotic diseases；The thrombotic diseases include But it is not limited to arterial thrombus and/or phlebothrombosis.

Platelet activating factor (Platelet activating factor, PAF) is a kind of with extensive bioactivity Endogenous phospholipid medium, with its receptor (PTAFR) combine to generate a series of receptor aftereffect.Under physiological status, Internal PAF is primarily involved in intracellular messengers and transmits and adjust, and the effect with reproduction, development of fetus, childbirth and kidney contacts closely. In pathological conditions, PAF can promote blood platelet, neutrophil accumulation, cell degranulation, release oxygen radical, tumour bad Necrosis factor, interleukin, arachidonic acid and metabolite promote thrombosis to increase vasopermeability, induce smooth muscle It shrinks, plays a significant role during a series of occurrence and development of diseases such as asthma, cardiovascular and cerebrovascular microcirculation disorder.Permitted in vivo Contain the paf receptor of specificity in many cells (such as endothelial cell, macrophage) surface.In a specific embodiment, the present invention adopts Inhibiting effect of the glabridin to PTAFR is detected with calcium current, the results showed that glabridin has obvious inhibiting effect to PTAFR.

Preferably, application of the present invention is to prepare the application in inhibiting platelet activating factor receptor drug.Light Licoricidin is used to inhibit a concentration of 50 μm of ol/L~200 μm ol/L of PTAFR；Preferably, a concentration of 150 μm of ol/L.

Ferric trichloride vasoactive damages blood vessel endothelium, further activates adherency, the aggregation capability of blood platelet.The present invention Influence of the glabridin to thrombus model platelet activity in two strains of rats is measured with gavage (oral) administering mode, the results showed that, light is sweet Grass is surely significant to reduce significant reduction FeCl₃Solution is applied with thrombus model rat suppository weight and blood plasma caused by arteria carotis Middle p- selectins, II b/ of vWF, GP III a and TXB2 are horizontal, and glabridin has anti thrombotic action.

Preferably, application of the present invention is to prepare platelet aggregation and activation caused by inhibiting blood vessel endothelium injury Application in drug.Wherein, the blood vessel endothelium injury is by FeCl₃Cause.

The present invention measures influence of the glabridin to rabbit hyperlipidemia thrombus model with intravenous administration approach, the results showed that, Glabridin can significantly inhibit the whole blood PAF concentration that Arterial thrombosis is induced and increase, and can significantly reduce IP in blood platelet₃ Content and blood platelet IP₃The expression of receptor shows glabridin to blood platelet PAF-IP₃-IP₃R-[Ca²⁺] i signal transduction pathways It is inhibited.

Preferably, application of the present invention is to prepare blood platelet PAF-IP₃-IP₃R-[Ca²⁺] i signal transduction pathways Application in drug.

The drug is using glabridin as agent is administered orally made of active ingredient and pharmaceutically acceptable auxiliary material Type, injecting medicine-feeding form, topical administration formulations.The dosage that glabridin treats thrombotic diseases is 10~40mg/kgd^-1, excellent It is selected as 40mg/kgd^-1。

The oral administered dosage form is tablet, capsule, granule, pill, oral solutions, soft extract, suspending agent, divides Powder, syrup.

The injecting medicine-feeding form is injection liquor or powder ampoule agent for injection.

The topical administration formulations are suppository, patch or gelling agent.

The tablet includes：Glabridin, starch, sodium carboxymethyl starch, talcum powder, dextrin, magnesium stearate and bonding Agent.Specifically, glabridin, starch, sodium carboxymethyl starch, talcum powder, dextrin and magnesium stearate mass ratio be 350:50: 7.5:0.8:50:0.8；Adhesive is 10% starch slurry.

The content of the capsule includes：Glabridin, starch, low-substituted hydroxypropyl cellulose, superfine silica gel powder, Magnesium stearate and adhesive.Specifically, glabridin, starch, low-substituted hydroxypropyl cellulose, superfine silica gel powder and magnesium stearate Mass ratio is 350:32:6:4.5:1.5；Adhesive is 10% starch slurry.

The granule includes：Glabridin, sucrose and dextrin.Specifically, the matter of glabridin, sucrose and dextrin Amount is than being 350:1000:500.

The pill includes：Glabridin, polyethylene glycol-6000 and Tween-80.Specifically, glabridin, The mass ratio of polyethylene glycol-6000 and Tween-80 is 350:12:80.5.

The injection liquor includes：Glabridin, soybean lecithin, glycerol and water.Specifically, glabridin, soybean Phosphatide, glycerine concentration be respectively 200g/L, 15g/L, 25g/L.When injection, liquor will be injected using 5% glucose injection After 250 times of dilution, intravenous drip.

Specific implementation mode

The present invention provides the purposes of glabridin, those skilled in the art can use for reference present disclosure, be suitably modified work Skill parameter is realized.In particular, it should be pointed out that all similar substitutions and modifications are aobvious and easy for a person skilled in the art See, they are considered as being included in the present invention.The method of the present invention and application are described by preferred embodiment, Related personnel can obviously not depart from the content of present invention, be modified to methods herein and application in spirit and scope or suitably It changes and combines, to realize and apply the technology of the present invention.

Drug, biomaterial or the instrument that the present invention uses are all common commercially available product, can all be bought in market.With reference to Embodiment, the technical solution that the present invention is further explained.

Inhibitory activity of 1 glabridin of embodiment to platelet activating factor receptor (PTAFR)

1, experiment material

1.1 cell strain：The Chinese hamster ovary cell strain CHO-K1 of stable transfection PTAFR, by Jiangsu Kang Yuan medicine company shares Co., Ltd is built using liposomal transfection teclmiques.

1.2 main agents：F12 culture mediums, tire ox blood (FBS) are purchased from Hyclone companies；Zeocin, HBSS are purchased from Invitrogen companies；Calcium current detection kit (6 dyestuff of calcium) is purchased from Molecular Device companies；PAF(C16), WEB2086 is purchased from tocris；HEPES is purchased from Nanjing Sheng Xing Bioisystech Co., Ltd；DMSO is purchased from sigma companies.

1.3 instrument：FLIPR tetra type microplate reader, Molecular Devices；Pipettor, eppendorf companies；It is super Net workbench is purchased from Shanghai Jie Jia air purifying process Co., Ltd；CDX-K1 inverted microscopes are purchased from Olympus； Countess Auto-counting of Cells instrument is purchased from invitrogen companies；3111 type cell incubators are purchased from Thermo Scientific companies.

2, experimental method

2.1 cell culture

Cell passage is inoculated in 10cm culture dishes, using complete medium (the F12 culture mediums containing 10%FBS), in 37 DEG C, 5%CO₂, saturated humidity incubator in cultivate, (cell is in logarithmic growth after the cell in culture dish covers with substantially Phase), original culture medium is discarded, addition 5mL PBS solutions, which are gently swung, washes, and discards, 0.25% trypsin solution is then added 0.5mL room temperatures digest 1 minute, and the digestion that the fresh complete mediums of 5mL terminate pancreatin is added, by cell be transferred to glass from In heart pipe, 800rpm centrifuges 4min, discards supernatant, and complete medium is added, careful piping and druming is at unicellular.Auto-counting of Cells Instrument meter number, it is about 3 × 10 to be then diluted to cell concentration with complete medium⁵A/mL is inoculated in 96 holes according to 100 μ L of every hole In the saturating tissue culture plate in black wall bottom, the 1st row are not added with cell and only add 100 μ L complete mediums, continue at 37 DEG C, 5%CO₂Culture Overnight incubation in case.

2.2 making up a prescription

The HEPES buffer solution of 1M is prepared, HBSS+20mM HEPES buffer solutions (hereinafter referred to as HBSS buffer solutions) are prepared. According to table 1, with HBSS+20mM HEPES buffer solutions dilution glabridin, positive drug WEB2086 and agonist PAF (C16).

2.3 detection

1. the cell of logarithmic growth phase digests for single cell suspension, counting.

2. cell is diluted to 3 × 10 with complete medium⁵A cell/mL is inoculated with 100 per hole in 96 cell culture plates μL.37 DEG C, 5%CO₂Overnight incubation.

3. being inhaled from tissue culture plate with the volley of rifle fire and abandoning 50 μ L cell culture fluids, 6 dyestuff of calcium that 50 μ L are preheated to room temperature is added. Culture plate is put into incubator and cultivates 30min.

4. use HBSS buffer solution diluted compounds, by after dilution compound and positive drug (WEB2086) be added cell training It supports in plate, 25 μ L sample solution is added per hole.Tissue culture plate is put into incubator and continues to cultivate 30min.

5. arriving 600nM with HBSS buffer solutions dilution agonist PAF (C16), it is added in 96 hole dispensing plates, per 80 μ L of hole, The HBSS blank controls for being not added with agonist are set.

6. the calcium current variation with FLIPR tetra detections per hole (is first scanned 2 minutes, is further continued for after 25 μ L agonists are added Scan 3min).

7. data export Δ RFU (i.e. RFU max-RFUmin in 0-300s).

3 experimental results

Inhibiting rate calculation formula：

Inhibiting rate %=1- [(sample sets-HBSS blank control groups)/(activator group-HBSS blank control groups)] × 100%

Table 1 is shown：Glabridin has platelet activating factor receptor certain inhibitory activity, and there are dose-dependant passes System.

Inhibitory activity of 1 glabridin of table to PTAFR

Influence of 2 glabridin of embodiment to thrombus model platelet activity in two strains of rats

1. experiment material

Cleaning grade SD rats 60, weight 180-220g or so, half male and half female are purchased from Beijing dimension tonneau China experimental animal Technology Co., Ltd.；II b/ of p-selectin, vWF, GP, III a and TXB2 kits are purchased from Shanghai and grind the limited public affairs of careful biotechnology Department；Remaining reagent is that domestic analysis is pure.

2. experimental method

2.1 experiment packets and modeling

60 SD rats routine feedings are randomly divided into 5 groups after a week：Sham-operation group, model group, glabridin be low, in, High dose group, every group each 12.Medication group is dissolved in physiological saline by 100,200,400mg/kg weight standards and carries out gavage light Licoricidin, sham-operation group and model group gavage normal saline, one time a day, continuous 7 days.Reference after last dose 30min Kurz methods replicate carotid thrombosis model.Blunt separation right carotid, it is beneath to pad 3cm²Medical dressing, remove sham-operation Remaining each group outside group is with being soaked with 20%FeCl₃The filter paper item of solution wraps up this section of arteria carotis communis, and sham-operation group physiology salt Water substitutes 20%FeCl₃Solution.Filter paper item is removed after 15min, dressing both ends blood vessel is ligatured after 40min, according to dressing length essence The blood vessel of filter paper item package, and removal of thromboses are really cut, is weighed.Rat heart takes blood, blood sodium citrate anti-freezing to prepare blood Slurry is used for the detection of blood plasma index.

2.2 blood plasma index determinings

With ELISA method detection p- selectins, the level of vWF, II b/ of platelet membrane glycoprotein GP, III a and TXB2, detection side Method is carried out according to kit specification.

2.3 statistical analysis

The data obtained result is with mean ± standard deviationIt indicates, it is for statistical analysis using SPSS16.0 softwares, it is more Comparison among groups use one-way analysis of variance, between group t inspections indicated with significant difference with P < 0.05.

3. experimental result

Influence of 3.1 glabridins to thrombus weight

As shown in table 2, sham-operation group is without thrombosis, and model group thrombus weight mean value is 3.4mg, illustrates that model group is made Mould success.Compared with model group, the thrombus weight of the middle and high dosage group of glabridin is substantially reduced, has significant difference, prompts Glabridin has anti thrombotic action.

2 glabridin of table to rat suppository weight influence (N=12)

Compared with model group, * P<0.05, * * P<0.01.

Influence of 3.2 glabridins to rat plasma index of correlation

Table 3 shows that p- selectins, II b/ of vWF, GPGP, III a and TXB2 levels are apparently higher than sham-operation group, this suggests that three Iron chloride vasoactive damages blood vessel endothelium later, further activates adherency, the aggregation capability of blood platelet.With model group phase Than p- selectins, II b/ of vWF, GP, the III a and TXB2 levels of middle and high dosage group decline to some extent, prompt glabridin logical The aggregation crossed the expression for inhibiting II b/ of blood platelet GPGP, III a and mitigate blood platelet, to reduce thrombosis.

3 glabridin of table to p- selectins, II b/ of vWF, GPGP, III a and TXB2 influence (N=12)

Influence of 3 glabridin of embodiment to rabbit hyperlipidemia thrombus model

1. experiment material

Tonneau China experimental animal is tieed up in male and healthy Japan white big ear rabbit 40, weight 2.2-2.5kg, 5-6 monthly age, Beijing Technology Co., Ltd..

2. experimental method

2.1 hyperlipemia models are established and grouping

Experimental rabbit single cage basal feed feeds 3d, and morning next day takes ear vein blood to look into serum total cholesterol (total on an empty stomach cholesterol,TC).The blood same day was taken to be fed with to 1% cholesterol, the daily 100g/kg of every rabbit.The ends 14d are tested, ear is taken Venous blood checks serum TC.If serum TC level is more than 5 times or more before feeding cholesterol, the success of rabbit hyperlipemia model is indicated. Above-mentioned 40 Hyperlipidemia Rabbits are randomly divided into 4 groups, every group 8：1. sham-operation group, does not replicate thrombus model, do not use up sweet Careless fixed intervention；2. model group replicates merely thrombus model, injecting normal saline；3. glabridin low dose group (50mg/kg), Glabridin 50mg/kg interventions are injected simultaneously replicating thrombus model；4. glabridin middle dose group (100mg/kg), is being replicated Thrombus model is injected glabridin 100mg/kg and is intervened simultaneously；5. glabridin high dose group (200mg/kg) is replicating thrombus Model is injected glabridin 200mg/kg and is intervened simultaneously.

2.2 anti-arterial platelet dependence thrombosis experiments

It is slightly improved by Ubatuba methods, prepares thrombotic model.Left carotid artery is detached, makes free segment length about Then 1.5cm injects isometric glabridin 50mg/kg, 100mg/kg, 200mg/kg and physiological saline from ear vein respectively, Each dosage group glabridin is all made of normal saline dissolving.After 30min is administered, with 2 stainless steel electrode (electrode diameters 1.5mm, spacing 2.0mm) free artery is gently provoked, with 2.0mA galvanic current stimulation 10min, then by semiconductor point temperature Degree meter places artery distal end, continuous to measure artery surface temperature.The time suddenly declined is stimulated to temperature since electric current i.e. The time (occlusive thrombus formation time, OT) is formed for occluding thrombus, shows that intra-arterial is formed at this time Platelet thrombus.5min after thrombosis takes blood 15mL to be detected for experiment from the puncture of the rabbit chambers of the heart.

2.3 whole blood PAF assays

5mL whole bloods are injected in advance immediately added with being shaken up in the test tube of equivalent methanol, are vibrated, centrifugation takes supernatant with having Solvent (chloroform：Methanol=1:2, V/V) extract lipid, take out chloroform phase, nitrogen be blown to it is micro, using high-performance thin layer chromatography (HPTLC) Measure PAF contents.

2.4 blood platelet IP₃Concentration mensuration

1 × 10⁹It is added in/ml platelet suspensions³H- phosphoinositides (IP₃) 1850kBq/mL, set 5%CO₂In incubator 3h is cultivated, 10mmol/L LiCl processing cells 15min, PBS wash cell, and chlorination is imitative：Methanol=1:2 stopped reactions simultaneously dissolve Cell.Cell solution water phase is taken, the miniature column precipitator of Dowex 1 × 8Formate types (Sigma) is packed into, collects IP₃Section eluent, Each pipe radioactivity of liquid flashing determining.

2.5 blood platelet IP₃R expression measures

The platelet concentration of separation is adjusted to 1 × 10⁸/ mL adds anti-IP respectively₃R monoclonal antibodies (LSBio, article No. LS- C138557), room temperature is protected from light lower standing 15min, adds IgG-FITC secondary antibodies (SouthernBiotech, article No. 4090-02) It is incubated 10min, flow cytomery IP3R protein expression intensity at room temperature.

2.6 statistical analysis

3, experimental result

3.1 glabridins are to IP in whole blood PAF, blood platelet₃Concentration and IP₃The influence of R expression

As shown in table 4, Arterial thrombosis can induce whole blood PAF concentration and significantly increase, IP in induced platelet₃Concentration is aobvious Write raising and IP₃R protein expressions dramatically increase.Glabridin intravenous administration can reduce PAF, IP in various degree₃And IP₃R's It is horizontal.

4 glabridin of table is to IP in PAF, blood platelet₃Concentration and IP₃R expression influence (N=8)

Compared with sham-operation group,^##P<0.01；Compared with model group, * * P<0.01.

Originally the experimental results showed that, glabridin has very strong anti-hyperlipidemia Arteries of Rabbits blood platelet dependence thrombus shape At effect.IP₃It is signal transduction substance important in blood platelet, IP₃-IP₃R-Ca²⁺The activation of release signal approach is blood platelet One of main mechanism of activation.PAF is the strongest platelet aggregation found so far, it is now recognized that PAF activates blood The mechanism of platelet is that blood platelet acyl inositol (IP) is promoted to decompose, IP in blood platelet₃Generation increases, by acting on IP₃R promotes Platelet dense pipe-line system Ca²⁺Release and Ca²⁺Miscarriage life, so as to cause platelet aggregation.The study show that glabridin Intravenous injection can significantly inhibit the whole blood PAF concentration that Arterial thrombosis is induced and increase, and can significantly reduce in blood platelet IP₃Content and blood platelet IP₃The expression of receptor.Therefore, it is considered that glabridin is to blood platelet PAF-IP₃-IP₃R-[Ca²⁺] i signals turn Approach inhibiting effect is led, is one of the mechanism of glabridin D-dimer and aggregation.

4 capsule of embodiment

By 350g glabridins and 32g starch, 6g low-substituted hydroxypropyl celluloses, 4.5g superfine silica gel powders, 1.5g stearic acid Magnesium and the mixing of appropriate 10% starch slurry, are packed into capsule, obtained capsule preparations 1000.3 times a day, 1 tablet each time.

5 granule of embodiment

350g glabridins and 1000g sucrose and 500g dextrin are mixed, 1000 packet granules are conventionally made. 3 times a day, 1 tablet each time.

6 tablet of embodiment

350g glabridins and 50g starch, 7.5g sodium carboxymethyl starches, 0.8g talcum powder, 50g dextrin, 0.8g is stearic Sour magnesium and the suitable mixing of appropriate 10% starch slurry, are conventionally made 1000, tablet.3 times a day, 1 tablet once.

6 pill of embodiment

350g glabridins and 12g polyethylene glycol-6000,80.5g Tween-80s, appropriate liquid paraffin are mixed, pressed Pill 1000 is made in more solito.3 times a day, 1 tablet each time.

7 injection of embodiment

By 200g glabridins and 15g injections soybean lecithin, 25g glycerol for injection, water for injection is settled to 1000mL, Injection 1000 is conventionally made.One time a day, every time 1, at least use 5% glucose injections of 250mL dilute Release rear intravenous drip.

It the above is only the preferred embodiment of the present invention, it is noted that those skilled in the art are come It says, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should be regarded as Protection scope of the present invention.

Claims

1. application of the glabridin in preparing the drug for treating or preventing thrombotic diseases.

2. application according to claim 1, it is characterised in that the application is to prepare inhibition platelet activating factor Application in receptor agents.

3. application according to claim 1, it is characterised in that the application is to inhibit blood vessel endothelium injury to draw in preparation Application in the platelet aggregation and pharmacological activation that rise；Wherein, the blood vessel endothelium injury is by FeCl₃Cause.

4. application according to claim 1, it is characterised in that the application is to prepare blood platelet PAF-IP₃-IP₃R- [Ca²⁺] application in i signal transduction pathway drugs.

5. application according to claim 1, it is characterised in that the drug is using glabridin as active ingredient and medicine Oral administered dosage form, injecting medicine-feeding form, topical administration formulations made of acceptable auxiliary material on.

6. application according to claim 5, it is characterised in that the oral administered dosage form is tablet, capsule, particle Agent, pill, oral solutions, soft extract, suspending agent, dispersant, syrup；The injecting medicine-feeding form is injection liquor or note It penetrates and uses powder-injection；The topical administration formulations are suppository, patch or gelling agent.

7. a kind of drug treating or preventing thrombotic diseases, it is characterised in that the drug is using glabridin as active ingredient.

8. drug according to claim 7, it is characterised in that the drug as active ingredient and pharmaceutically may be used using glabridin Oral administered dosage form, injecting medicine-feeding form or topical administration formulations are made in the auxiliary material of receiving.

9. drug according to claim 8, it is characterised in that the oral administered dosage form is tablet, capsule, particle Agent, pill, oral solutions, soft extract, suspending agent, dispersant, syrup；

The injecting medicine-feeding form is injection liquor or powder ampoule agent for injection；