Plant extract composition as well as preparation method and application thereof

Abstract

The invention relates to the technical field of plant extracts. The invention provides a plant extract composition as well as a preparation method and application thereof. The plant extract composition comprises a ligusticum wallichii extract and an olive extract, the mass ratio of the ligusticum wallichii extract to the olive extract is (10-0.5): (0.01-0.5). The rhizoma chuanxiong extract and the olive extract are combined for use, so that a good synergistic interaction effect is achieved, and a good treatment effect is achieved on thrombosis, particularly venous thrombosis. And the acute toxic effect is not increased.

Description

Technical Field

[0001] The present invention relates to the technical field of plant extracts, in particular to a plant extract composition, a preparation method and application thereof. Background Art

[0002] Thrombotic diseases mainly include arterial thrombotic diseases, venous thrombotic diseases and capillary thrombotic diseases. Common arterial thrombotic diseases include stroke, coronary heart disease and angina pectoris; venous thrombotic diseases mainly include lower limb venous thrombosis, deep vein thrombosis, and pulmonary embolism and cerebral embolism formed after thrombus detachment. The formation of thrombus is mainly related to changes in blood vessel walls, blood components and hemodynamics.

[0003] Venous thrombi are mainly composed of fibrin and red blood cells. Venous thrombi are formed under low shear conditions, a process that can occur from direct damage to the vessel wall during surgery, after trauma, or triggered by mechanical damage secondary to an indwelling intravenous catheter. Typically, venous thrombi originate from the deep veins of the calf, and proximal venous thrombi are more likely to dissociate and embolize to the lungs, forming pulmonary embolism, compared with calf vein thrombi.

[0004] Damage to vascular endothelial cells can cause platelet activation and aggregation, activate the endogenous and exogenous coagulation systems, convert fibrinogen into fibrin, and then entrap a large number of red blood cells, leading to thrombosis. Vascular endothelial cells are a layer of flat epithelial cells in the inner layer of the cardiovascular system. They can synthesize and secrete a variety of bioactive substances such as endothelin (ET), adenosine diphosphatase, vascular endothelial growth factor, nitric oxide, etc. They have many important physiological functions: providing a place for the synthesis of prostacyclin; converting adenosine diphosphate into adenine nucleotides to inhibit platelet adhesion; activating the protein C system, inhibiting coagulation factors Va and VⅢa, thereby producing an anticoagulant effect; it can be seen that the formation and degradation of thrombi in the body involve multiple links, and the influencing factors are very complex and diverse.

[0005] In order to minimize the damage of thrombosis to the body, a variety of drugs for preventing and treating thrombosis have been developed for different links of the coagulation process, including antiplatelet aggregation drugs such as low-dose aspirin; anticoagulants such as warfarin, and thrombolytic drugs such as viper antithrombin. These drugs are difficult to control in dosage and have a high risk of causing bleeding. A variety of new antithrombotic drugs have been developed to address the above adverse reactions, and some have entered the clinical trial stage. For example, Revacept, a platelet adhesion inhibitor composed of the extracellular domain of the GPVI receptor and the human Fc-fragment, has entered clinical phase II, mainly by inhibiting collagen-induced platelet aggregation to exert its antithrombotic effect; among anticoagulants, the endogenous coagulation inhibitor FIXa monoclonal antibody (SB249417) and FIXa oral inhibitor (TTP889) have also entered the clinical trial stage, but their safety and effectiveness also need further study.

[0006] Chuanxiong is the dried rhizome of the plant Chuanxiong Ligusticumchuanxiong Hort. of the Umbelliferae family. It is pungent and warm. It enters the liver, gallbladder and pericardium meridians; it has the effects of promoting blood circulation and qi, dispelling wind and relieving pain. It is widely used clinically for chest pain, chest and flank pain, swelling and pain due to falls, irregular menstruation, amenorrhea and dysmenorrhea, abdominal pain due to lumps, headache, and rheumatic pain. It is a famous authentic Chinese medicinal material produced in Sichuan. Modern research shows that Chuanxiong mainly contains organic acids (representative components are ferulic acid, chlorogenic acid and caffeic acid, etc.), phthalide compounds (representative components such as ligustilide, ligustilide A, etc.), alkaloids (such as ligustrazine) and polysaccharides. In the research on Chuanxiong, people have focused on water-soluble components such as ferulic acid and ligustrazine, but less attention has been paid to its volatile components. The reason is related to the loss of more effective components in the volatile oil obtained previously. Although traditional Chinese medicine lists Chuanxiong as a drug for promoting blood circulation and removing blood stasis, most people believe that its active ingredient should be its water-soluble component because it is routinely used in clinical practice as a decoction.

[0007] Olea europare L is a plant of the genus Olea in the family Oleacea. It is native to Asia Minor and is now widely distributed in more than 30 countries around the world. my country began to introduce it in the 1950s and 1960s, and it is now widely planted in Sichuan, Yunnan, Gansu and other places. According to statistics, in 2013, the national olive planting area reached 570,000 mu; by 2020, the national planting area has reached 80,000 hectares (about 1.2 million mu). With the development of the olive industry, the output of its byproduct olive leaves has also increased year by year. According to statistics, Gansu alone produces 1,200 tons of olive leaves each year. However, due to the weak research and development capabilities of olive leaves, it may become a potential risk factor that harms the environment and ecology. In-depth research has found that olive leaves contain a variety of active substances, including: polyphenols, flavonoids, terpenes, aromatic compounds, aliphatic compounds and other chemical substances. It has multiple pharmacological effects such as antiviral, antibacterial, anti-tumor, antioxidant, anti-stroke, analgesic, and anti-inflammatory.

[0008] Therefore, there is an urgent need for antithrombotic drugs with good efficacy and few adverse reactions in clinical practice. Based on the stability of the effective substances in Chuanxiong extract in the early stage, our team further discovered through a series of innovative studies that Chuanxiong extract and olive extract have obvious synergistic effects in the field of preventing and treating thrombosis, and are highly safe. Summary of the invention

[0009] In view of this, the technical problem to be solved by the present invention is to provide a plant extract composition, which can synergistically inhibit thrombosis.

[0010] The invention provides a plant extract composition, comprising a rhizoma chuanxiong extract and an olive extract; the mass ratio of the rhizoma chuanxiong extract to the olive extract is 10-0.5:0.01-0.5.

[0011] Preferably, in the Chuanxiong extract, ligustilide ≥5.0% and ligustilide A ≥3.0%.

[0012] Preferably, in the olive extract, oleuropein is ≥ 50.0%.

[0013] Preferably, the preparation method of the Chuanxiong extract comprises: ethanol reflux extraction, supercritical carbon dioxide extraction or subcritical fluid extraction;

[0014] The ethanol reflux extraction comprises: heating chuanxiong and 70% ethanol for reflux extraction, and performing reduced pressure distillation to obtain the product;

[0015] The supercritical carbon dioxide extraction comprises: extracting Chuanxiong with CO2 gas to obtain;

[0016] The subcritical fluid extraction comprises: extracting Chuanxiong with butane subcritical fluid to obtain the product.

[0017] Preferably, the preparation method of the olive extract comprises:

[0018] The olive leaves and solvent are mixed, extracted, and the filtrate is passed through a macroporous resin to obtain the product; the solvent includes ethanol or petroleum ether; the ethanol is a 60% to 70% ethanol aqueous solution.

[0019] The present invention provides a method for preparing a plant extract composition, comprising:

[0020] The product is obtained by mixing the chuanxiong extract and the olive extract.

[0021] The present invention provides use of any one of the plant extract compositions described above in the preparation of a medicament for preventing and / or treating thrombosis.

[0022] The present invention provides a drug for preventing and / or treating thrombosis, comprising the plant extract composition described in any one of the above technical solutions.

[0023] Preferably, the dosage form of the drug includes, but is not limited to, one or more of granules, decoctions, powders, oral liquids, tablets, pills, pastes or capsules.

[0024] Preferably, the prevention and / or treatment of thrombosis includes one or more of inhibiting the formation of thrombus, reducing the weight of thrombus or reducing the length of thrombus.

[0025] Compared with the prior art, the present invention provides a plant extract composition, including Chuanxiong extract and olive extract; the mass ratio of the Chuanxiong extract to the olive extract is 10-0.5:0.01-0.5. The Chuanxiong extract and the olive extract of the present invention are used together, which has a good synergistic effect and has a good therapeutic effect on thrombosis, especially venous thrombosis. And its acute toxicity is not increased. DETAILED DESCRIPTION

[0026] The present invention provides a plant extract composition, a preparation method and application thereof. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve the above. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and they all fall within the scope of protection of the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0027] In this application, the term "and / or" describes the association relationship of associated objects, indicating that there may be three relationships. For example, A and / or B can represent: A exists alone, A and B exist at the same time, and B exists alone. A and B can be singular or plural.

[0028] In this application, "at least one" means one or more, "more than one" means two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single or plural items.

[0029] The present invention provides a plant extract composition, comprising a chuanxiong extract and an olive extract; the mass ratio of the chuanxiong extract to the olive extract is 10-0.5:0.01-0.5, and the best preferred range is 10:0.01-0.7.

[0030] The plant extract composition provided by the invention comprises chuanxiong extract and olive extract.

[0031] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 10:0.01-0.7.

[0032] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 10:0.01-0.5.

[0033] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 10:0.05-0.7.

[0034] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 10:0.01.

[0035] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 10:0.05.

[0036] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 10:0.5.

[0037] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 10:0.7.

[0038] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 0.5-2:0.05-0.2;

[0039] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 0.5:0.05 to 2.0:0.05.

[0040] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 0.5:0.05;

[0041] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 2:0.05;

[0042] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 0.5:0.2;

[0043] In some preferred embodiments of the present invention, the mass ratio of the Chuanxiong extract to the olive extract is 2:0.2;

[0044] According to the present invention, the preparation method of the Chuanxiong extract comprises: ethanol reflux extraction, supercritical carbon dioxide extraction or subcritical fluid extraction.

[0045] The ethanol reflux extraction of the present invention comprises: heating chuanxiong and 70% ethanol for reflux extraction, and distilling under reduced pressure to obtain. More preferably, it comprises: soaking chuanxiong and 7 to 9 times of 70% ethanol for 10 to 12 hours, then heating and refluxing for 2 hours, filtering, and obtaining medicinal residues; adding 3 to 5 times of 70% ethanol to the medicinal residues, heating and refluxing for 2 hours, combining the filtrate, and distilling under reduced pressure to obtain chuanxiong extract. The percentage of ethanol in the present invention refers to the volume percentage.

[0046] In a specific embodiment, Chuanxiong is crushed into coarse powder, added with 8 times the amount of 70% ethanol, soaked for 12 hours, heated under reflux for 2 hours, and filtered; 4 times the amount of 70% ethanol is added to the residue, heated under reflux for 2 hours, the filtrate is combined, and vacuum distillation is performed to obtain Chuanxiong extract.

[0047] In the chuanxiong extract of the present invention, ligustilide is ≥5.0% and ligustilide A is ≥3.0%.

[0048] The supercritical carbon dioxide extraction of the present invention comprises: extracting rhizoma chuanxiong with CO2 gas to obtain the product.

[0049] More preferably, the method specifically comprises: adding 7 to 9 times of CO2 gas to extract Chuanxiong.

[0050] The extraction conditions are: temperature 45°C ~ 55°C, pressure 28 ~ 35MPa, time 120min, CO2 flow rate 18-25L / min, separate the residue, extract the residue again with the same method, and combine the two extracts.

[0051] The supercritical carbon dioxide extraction of the present invention is preferably carried out in a complete set of supercritical extraction equipment, which is not limited in the present invention.

[0052] The subcritical fluid extraction of the present invention comprises: extracting Chuanxiong with butane subcritical fluid to obtain the product.

[0053] The subcritical fluid of the present invention is preferably extracted in a subcritical extraction complete set of equipment, but the present invention is not limited thereto.

[0054] Compress butane gas into a subcritical fluid through an air compressor (critical pressure: 0.47Mpa);

[0055] The preferred embodiment of the present invention specifically comprises: introducing a quantitative amount of butane subcritical fluid (mass-to-volume ratio is 1:8) into an extraction tank; extracting at 45° C. for 55 minutes; filtering the extract from the bag until all the extract is exhausted, and guiding the extract into a desolventizing tank; extracting twice according to the above method, and combining the extracts.

[0056] The desolventizing tank is gradually depressurized to normal pressure to completely vaporize the butane in the extract and recover the butane; after the temperature in the tank drops to room temperature, the degassing is performed under reduced pressure. The extract is collected.

[0057] The preparation method of the olive extract of the present invention comprises:

[0058] The olive leaves are mixed with a solvent, extracted, and the filtrate is passed through a macroporous resin to obtain the product.

[0059] According to the present invention, the solvent includes ethanol or petroleum ether; the ethanol is a 60% to 70% ethanol aqueous solution.

[0060] In a specific embodiment of the present invention, olive leaves and 60% ethanol are refluxed for extraction, the reflux temperature is 75° C. to 85° C., the extraction time is 2 hours, and the extraction times are 1 to 3 times, preferably 2 times.

[0061] The mass volume ratio of the olive leaves to 60% ethanol is 1:4-8.

[0062] The combined extracts were concentrated, the concentrate was loaded onto a macroporous resin column by wet method, the column diameter-to-height ratio was 1:8, the volume and flow rate of the loading solution were 10 BV and 2 BV / h, the volume and flow rate of the deionized water elution were 10 BV and 10 BV / h, the eluent was 60% ethanol solution, the volume and flow rate were 8 BV and 3 BV / h, the eluent was collected, concentrated, and dried to obtain the extract.

[0063] In the olive extract of the present invention, oleuropein is ≥50.0%.

[0064] The present invention provides a method for preparing a plant extract composition, comprising:

[0065] The product is obtained by mixing the chuanxiong extract and the olive extract.

[0066] The preparation methods of the chuanxiong extract and the olive extract have been clearly described above in the present invention, and will not be repeated here.

[0067] The present invention provides use of any one of the plant extract compositions described above in the preparation of a medicament for preventing and / or treating thrombosis.

[0068] The present invention provides a drug for preventing and / or treating thrombosis, comprising the plant extract composition described in any one of the above technical solutions.

[0069] The drug for preventing and / or treating thrombosis provided by the present invention can be mixed with commonly used auxiliary materials in pharmacy to prepare an oral preparation that is convenient to take.

[0070] The dosage form of the drug of the present invention includes, but is not limited to, one or more of granules, decoctions, powders, oral liquids, tablets, pills, pastes or capsules.

[0071] The medicine of the present invention also includes conventional auxiliary materials used by those skilled in the art, which are not limited here.

[0072] In some embodiments provided by the present invention, the pharmaceutically acceptable excipient is one or a mixture of two or more of fruit powder, edible flavor, sweetener, acidulant, filler, lubricant, preservative, suspending agent, edible coloring, diluent, emulsifier, disintegrant or plasticizer.

[0073] Fillers refer to auxiliary materials used to increase the weight and volume of tablets and facilitate tableting. Fillers used as tablets require good fluidity and compressibility, strong binding force, and a large capacity for drugs. Preferably, the filler is one or a mixture of two or more of fructose, sugar alcohols, sucrose, starch, pregelatinized starch, microcrystalline cellulose or dextrin. However, those skilled in the art believe that feasible disintegrants are within the scope of protection of the present invention, and the types of fillers are not limited thereto, and the present invention is not limited thereto.

[0074] Lubricant refers to an auxiliary material that can reduce the friction between the tablet and the die wall, so as to prevent the friction from being large and making tableting difficult, and can make the pressure distribution during tableting uniform, and make the density of the tablet uniform, and can also improve the appearance of the tablet, making the tablet surface bright and smooth. Preferably, the lubricant is one or a mixture of two or more of talcum powder, magnesium stearate, silicon dioxide or stearic acid. However, those skilled in the art believe that feasible lubricants are all within the scope of protection of the present invention, and the type of lubricant is not limited thereto, and the present invention is not limited thereto.

[0075] The main function of the disintegrant is to eliminate the binding force of the tablet formed by the adhesive or by pressure, so that the tablet disintegrates rapidly and the tablet can quickly exert its medicinal effect. Preferably, the disintegrant is one of dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, cross-linked polyvinyl pyrrolidone or sodium carboxymethyl cellulose, or a mixture of two or more thereof. However, those skilled in the art believe that feasible disintegrants are within the scope of protection of the present invention, and the types of fillers are not limited thereto, and the present invention is not limited thereto.

[0076] The prevention and / or treatment of thrombosis in the present invention includes one or more of inhibiting the formation of thrombus, reducing the weight of thrombus or reducing the length of thrombus.

[0077] The present invention provides a plant extract composition, comprising a chuanxiong extract and an olive extract; the mass ratio of the chuanxiong extract to the olive extract is 10:0.01-0.5. The chuanxiong extract and the olive extract of the present invention are used in combination, which has a good synergistic effect and has a good therapeutic effect on thrombosis, especially venous thrombosis. However, the acute toxicity thereof is not increased.

[0078] It should be understood that in the various embodiments of the present application, the size of the serial numbers of the above-mentioned processes does not mean the order of execution, some or all of the steps can be executed in parallel or sequentially, and the execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0079] The numerical ranges and parameters involved in the present invention have presented the relevant numerical values ​​in the specific embodiments as accurately as possible. However, any numerical value essentially inevitably contains standard deviations due to individual testing methods. Therefore, unless otherwise explicitly stated, it should be understood that all numerical ranges or specific data used in this disclosure may have certain reasonable deviations within a certain range, for example: within 1% or 0.5%.

[0080] Some cases are recorded in the embodiments and comparative examples of the present invention, wherein the embodiments show certain implementation modes of the present invention. However, this does not mean that the effects of the present invention can only be achieved in these cases.

[0081] In order to further illustrate the present invention, a plant extract composition, a preparation method and application thereof provided by the present invention are described in detail below in conjunction with examples.

[0082] In the tables of the embodiments of the present invention, x±S refers to “mean±standard deviation” well known to those skilled in the art.

[0083] Example 1 Ethanol Extraction of Ligusticum chuanxiong Volatile Oil

[0084] Take 1.0kg of Chuanxiong, crush it into coarse powder, add 8 times the amount of 70% ethanol, soak for 12 hours, heat and reflux for 2 hours, filter; add 4 times the amount of 70% ethanol to the residue, heat and reflux for 2 hours, combine the filtrate, and use vacuum distillation to obtain Chuanxiong volatile oil. The yield is 4.3%, containing 5.0% ligustilide and 3.0% ligustilide A.

[0085] Example 2 Supercritical extraction of volatile oil from Ligusticum chuanxiong

[0086] Take 1.0kg of Chuanxiong, crush it into coarse powder, add it to the supercritical extraction equipment, put it into the extraction tank, and seal the tank mouth. Add 8 times the amount of CO2 gas for extraction. The extraction conditions are: temperature 45℃~55℃, pressure 28~35MPa, time 120min, CO2 flow rate 18-25L / min, separate the residue, extract the residue again with the same method, combine the two extracts, recover CO2, and the yield is about 5.4%. It contains 5.5% of ligustilide and 3.6% of ligustilide A.

[0087] Example 3 Subcritical extraction of volatile oil from Ligusticum chuanxiong

[0088] Take the crude powder of Chuanxiong, put it into a clean cloth bag, 1.0kg per bag, and tie it. Place it in the extraction tank of the subcritical fluid extraction equipment and seal the tank mouth. Compress butane gas through an air compressor (critical pressure: 0.47Mpa) into a subcritical fluid and place it in a storage tank for use. Introduce a certain amount of butane subcritical fluid (mass volume ratio is 1:8) into the extraction tank; extract at 45℃ for 55 minutes; lift the material frame until the extract is completely filtered out of the cloth bag, and guide the extract into the desolventizing tank; extract twice according to the above method and combine the extracts. Gradually depressurize the desolventizing tank to normal pressure to completely vaporize the butane in the extract and recover the butane; after the temperature in the tank drops to room temperature, decompress and degas. Collect the extract, weigh it, and calculate the yield to be 7%. This product contains ligusticum chuanxiong lactone A (C 12 H 16 O2) 5.0%, containing ligustilide (C 12 H 14 O2)7.0%.

[0089] Example 4 Ethanol Extraction of Olive Leaf Extract

[0090] Weigh 1.0kg of olive leaves, add 12L of 60% ethanol solution, reflux extract for 2h, pour out the extract, add 10L of 60% ethanol solution, reflux extract for 2h, filter, combine the two extracts, concentrate to 10L, and use the concentrate as the loading solution. Take a macroporous resin wet column, the column diameter-to-height ratio is 1:8, the volume and flow rate of the loading solution are 10BV and 2BV / h respectively, the deionized water elution volume and flow rate are 10BV and 10BV / h respectively, the eluent is 60% ethanol solution, the elution volume and flow rate are 8BV and 3BV / h respectively, collect the eluent, concentrate, and dry to obtain an extract with a yield of 12% and oleuropein content>60.0%.

[0091] Example 5 Ethanol Extraction of Olive Leaf Extract

[0092] Take 1.0 kg of olive leaves (washed and dried), crush into coarse powder, add 8 times the amount of 70% ethanol, soak for 12 hours, heat and reflux (reflux temperature is 75℃~85℃) for 2 hours, filter; add 4 times the amount of ethanol to the residue, heat and reflux (reflux temperature is 75℃~85℃) for 2 hours, combine the filtrate, and pass through macroporous resin. The yield is 2.2%, containing 50% oleuropein.

[0093] Verification Example

[0094] 1. Experimental Materials

[0095] 1.1 Experimental drugs and reagents

[0096] The sample preparation for each example is shown above, and the sample preparation for animal experiments is shown in Table 1

[0097] Table 1 Composition of samples used in animal experiments

[0098]

[0099] Rivaroxaban tablets, 10 mg, batch number: BJ67929, produced by Bayer HealthCare Co., Ltd.

[0100] Sodium carboxymethyl cellulose, batch number: 2014102701AR, produced by Chengdu Kelong Chemical Reagent Factory.

[0101] Ferric chloride, hexahydrate (FeCl3), 500g, content ≥98.0%, batch number: 2022090101, Chengdu Kelong Chemical Co., Ltd.

[0102] Lipopolysaccharide (LPS), 100 mg, content ≥ 98.0%, batch number: 0000189842, produced by Sigma

[0103] 1.2 Experimental instruments

[0104] Analytical balance: Sartorius, Germany, model BP211D, e=0.1 mg, d=0.01 mg.

[0105] 1.4 Experimental animals and environment

[0106] 1.4.1 Experimental mice

[0107] SPF-grade male SD rats, weighing 150-180 g and 200-220 g, were purchased from the Experimental Animal Center of Sichuan Academy of Chinese Medicine, with production license number SCXK(Chuan)2018-19; they were all kept in the SPF barrier system of the Institute of Experimental Animal Center of Sichuan Academy of Chinese Medicine, with license number SYXK(Chuan)2018-100, with an indoor temperature of 20-22 °C, a relative humidity of 40%-70%, and 12 h alternating light and dark lighting.

[0108] 2. Experimental Methods

[0109] 2.1 Orthogonal design to explore the synergistic effect between Chuanxiong volatile oil and olive extract

[0110] 72 healthy male SD rats weighing 200-220g were selected and designed with two factors and three levels. The specific dosing regimen is shown in Table 2. The drug was administered twice a day for a total of 7 times. 40 minutes after the last administration, the rats were anesthetized and fixed in the supine position, the neck was depilated and disinfected, the midline incision of the neck was made, and the left common carotid artery was separated by 3cm. Stimulating electrodes were placed at both ends of the blood vessel (positive electrode at the proximal end and negative electrode at the distal end), with a 1cm interval between the stimulating electrodes, and a laser Doppler probe was placed between the two electrodes to measure the blood flow before and after stimulation. The laser Doppler was used to measure the blood flow before electrical stimulation. After stabilization, the blood vessel was stimulated with direct current (stimulation intensity 4mA) to start timing. When the blood flow dropped to 1 / 4 of the blood flow before electrical stimulation, the timing was stopped and the time taken was recorded. The time displayed from the start to the stop of the timing is the arterial thrombosis time.

[0111] As shown in Table 2, thrombosis can be formed in the common carotid artery of rats under electrical stimulation. The Chuanxiong extract of Example 3 and the combination of the Chuanxiong extract of Example 3 and the olive extract of Example 4 in different proportions can significantly prolong the time of thrombosis formation in the carotid artery of rats under electrical stimulation, and the difference is statistically significant compared with Experiment No. 1.

[0112] Table 2 Orthogonal design explores the effects of different proportions of Example 3 and Example 4 on electrical stimulation of arterial thrombosis

[0113]

[0114] Compared with the model group: *P<0.05; **P<0.01; ***P<0.001.

[0115] In order to further clarify the role of each drug in the prescription in thrombosis, as well as the interaction between each drug (antagonism, enhancement, synergy, etc.), the project team used SPSS statistical software (SPSS Statistics V26.0) to perform orthogonal design single-factor variance analysis on the original data in Table 2. When the software running results show that the significant data is less than 0.05, it indicates that the substance plays an important role in this effect; when the significant data is less than 0.01, it indicates that the substance plays a very important role in this effect;

[0116] Table 3 Example 3, Example 4 Interaction orthogonal design variance analysis table between-subject effect test, dependent variable: coagulation time

[0117]

[0118]

[0119] aR square = 0.588 (adjusted R square = 0.515)

[0120] From the orthogonal design analysis of variance table in Table 3, it can be seen that in prolonging the carotid artery thrombosis of rats with electrical stimulation, both Chuanxiong and olive extracts played a very important or important role (P<0.001 or P<0.05). When the two were used in combination, there was a significant synergistic effect between Chuanxiong and olive extracts in prolonging the carotid artery thrombosis time of rats (P<0.05).

[0121] 2.2 Effects of various examples on suspension-induced inferior vena cava thrombosis in rats

[0122] Male SD rats weighing 260-280g were randomly divided into 6 groups and given oral administration of each drug as shown in Table 1. The model group was given an equal volume of solvent edible oil by oral administration. The drugs were given by oral administration twice a day for a total of 7 times. 30 minutes after the third administration, the animals were anesthetized, the abdomen was opened, and the 1.0 cm long inferior vena cava segment below the confluence of the left renal vein and the inferior vena cava was separated. A "0" silk thread was passed below the confluence of the left renal vein into the inferior vena cava, and the two ends of the ligature were passed through the right side of the abdominal wall and led out of the abdominal cavity. In the same way, a ligature was placed above the common iliac vein. The intestine and its mesentery were returned to the abdominal cavity, and the position of the ligature was adjusted to avoid compressing the intestine. The two ligatures were tied and fixed outside the abdominal wall, 2.0 mL of normal saline was injected into the abdominal cavity, and the abdomen was closed. The animals were fed normally after surgery. The drug was continued for 4 times, and the animals were anesthetized 1 hour after the last administration. The embolized segment of the blood vessel was taken, and the residual blood was absorbed with filter paper, and the length and weight of the thrombus were measured respectively.

[0123] As can be seen from Table 4, 48 hours after the suspension-induced inferior vena cava thrombosis in rats, the inferior vena cava thrombosis was obvious, and the thrombus weight and thrombus length were statistically significant compared with the control group; each embodiment had a good inhibitory effect on the formation of venous thrombosis in rats induced by suspension surgery, among which Examples 3, 4, and 5 had the best effect.

[0124] Table 4 Effects of various examples on suspension-induced inferior vena cava thrombosis in rats

[0125]

[0126] Compared with the model group: *P<0.05; **P<0.01; ***P<0.001.

[0127] 2.3 Effects of each example on LPS+FeCl3-induced inferior vena cava thrombosis in rats

[0128] Male SD rats with a healthy body weight of 240g-260g were randomly divided into groups and given drugs by gavage as shown in Table 2. The model group was given an equal volume of solvent edible oil by gavage. Twice a day, a total of 7 times. The drug was given three times before modeling. 30 minutes after the third drug administration, except for the control group, the other groups were intraperitoneally injected with saline 3.0mg / kg LPS. 4 hours later, 10% chloral hydrate 350mg·kg was intraperitoneally injected. -1 Anesthetize the rats, make a midline incision in the abdomen to expose the inferior vena cava, separate the surrounding tissues from the inferior vena cava segment from the infrarenal vein to the iliac lumbar vein, place a small piece of plastic film (5cm*7mm) under the separated inferior vena cava, and cover the surface of the segment with a filter paper of about 6mm*6mm soaked in 6% FeCl3 solution for 3 minutes. Remove the filter paper after 3 minutes. Suture the abdomen, keep the animals warm after surgery, and eat and drink freely. Continue to give the drug 4 times, anesthetize the animals 48 hours later, remove the blood vessel part with thrombosis, measure the length and weight of the thrombus, and take blood from the abdominal aorta for four coagulation tests and renal function tests.

[0129] As can be seen from Table 5, 48 hours after LPS+FeCl3-induced inferior vena cava thrombosis in rats, the inferior vena cava thrombosis was obvious, and the thrombus weight and thrombus length were statistically significant compared with the control group; each embodiment had a good inhibitory effect on the formation of LPS+FeCl3-induced venous thrombosis in rats, among which Examples 3, 4, and 5 had the best effect.

[0130] Table 5 Effects of various examples on LPS+FeCl3-induced inferior vena cava thrombosis in rats

[0131]

[0132] Compared with the model group: *P<0.05; **P<0.01; ***P<0.001.

[0133] From the experiment 2.1, it can be seen that Chuanxiong extract can significantly prolong the time of thrombosis in the electrically stimulated carotid artery of rats, and olive extract also has a similar but weaker anti-thrombotic effect. When the two are used in combination, they have a significant synergistic effect. This anti-thrombotic effect has been further confirmed in different animal models such as 2.2 and 2.3.

[0134] In summary, putting Chuanxiong extract and olive extract together in a certain proportion has a synergistic preventive and therapeutic effect on thrombosis caused by various reasons, and is expected to become an ideal innovative new drug for the prevention and treatment of thrombotic diseases, meeting the clinical treatment options of patients.

[0135] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A plant extract composition, characterized in that The invention is composed of a rhizoma chuanxiong extract and an olive extract; the mass ratio of the rhizoma chuanxiong extract to the olive extract is 10-0.5:0.01-0.5; The preparation method of the Chuanxiong extract comprises: ethanol reflux extraction, supercritical carbon dioxide extraction or subcritical fluid extraction; The ethanol reflux extraction comprises: heating chuanxiong and 70% ethanol for reflux extraction, and performing reduced pressure distillation to obtain the product; The supercritical carbon dioxide extraction comprises: extracting Chuanxiong with CO2 gas to obtain; The subcritical fluid extraction comprises: extracting Chuanxiong with butane subcritical fluid to obtain; The preparation method of the olive extract comprises: The olive leaves and solvent are mixed, extracted, and the filtrate is passed through a macroporous resin to obtain the product; the solvent includes ethanol or petroleum ether; the ethanol is a 60% to 70% ethanol aqueous solution.

2. The plant extract composition according to claim 1, characterized in that In the chuanxiong extract, ligustilide≥5.0% and ligustilide A≥3.0%.

3. The plant extract composition according to claim 1, characterized in that In the olive extract, oleuropein is ≥50.0%.

4. A method for preparing the plant extract composition according to any one of claims 1 to 3, characterized in that: include: The product is obtained by mixing the chuanxiong extract and the olive extract.

5. Use of the plant extract composition according to any one of claims 1 to 3 in the preparation of a medicament for preventing and / or treating thrombosis.

6. A drug for preventing and / or treating thrombosis, characterized in that: A plant extract composition comprising any one of claims 1 to 3.

7. The drug according to claim 6, characterized in that The dosage form of the drug includes, but is not limited to, one or more of granules, decoctions, powders, oral solutions, tablets, pills, pastes or capsules.

8. The drug according to claim 6, characterized in that The prevention and / or treatment of thrombosis includes one or more of inhibiting the formation of thrombus, reducing the weight of thrombus or reducing the length of thrombus.

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