KR102672932B1 - Composition for prevention or treatment of thrombotic diseases, comprising an extract of Aster scaber as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for treating or preventing thrombotic diseases using an extract of Aster scaber . The Aster scaber extract effectively inhibits aggregation activity and thrombus formation in platelets induced by collagen, thrombin, and ADP, It acts as an effective inhibitor of blood clots or platelet aggregation and can be usefully used to prevent or treat various thrombotic diseases.

Description

Composition for prevention or treatment of thrombotic diseases, comprising an extract of Aster scaber as an active ingredient}

The present invention relates to a composition for preventing or treating thrombotic diseases containing Aster scaber (AS) extract as an active ingredient.

In modern society, traditional eating habits are threatened due to improvements in living standards and changes in lifestyle due to industrialization, and due to these changes in eating habits, various adult diseases such as blood-related diseases such as obesity, high blood pressure, and hyperlipidemia are rapidly increasing.

As a component of the human body, blood has a variety of important functions, including transport of oxygen, nutrients, and waste, buffering action, maintenance of body temperature, regulation of osmotic pressure and maintenance of ionic balance, maintenance of moisture content, fluid regulation, maintenance and control of blood pressure, and biological defense. there is. Therefore, blood and blood circulation are essential for human survival, and when blood vessels are damaged, blood loss must be prevented by quickly forming blood clots. Blood has parallel coagulation and dissolution reactions, and no blood clots (blood clots) are formed during normal blood circulation. However, when blood clots are formed due to external blood vessel damage, internal blood coagulation factors, and an imbalance of thrombolytic factors, blood vessels become blocked and blood circulation is interrupted. In this case, the important functions of the blood do not work, causing serious damage to the human body.

The blood coagulation reaction occurs when platelets adhere and aggregate to the blood vessel wall to form a platelet thrombus due to blood vessel damage or an imbalance of internal blood coagulation factors/dissolution factors, and then the blood coagulation system is activated to form a fibrin thrombus centered on the platelet aggregate. At this time, thrombin is activated through several stages of reaction of numerous blood coagulation factors, producing fibrin monomers from fibrinogen. The fibrin monomers are polymerized by calcium, bind to platelets and endothelial cells, and are cross-linked by factor XIII. By forming fibrin polymers, a permanent blood clot is created.

Excessive formation of blood clots within blood vessels interferes with blood circulation and, in severe cases, blocks blood vessels, causing serious damage to the cardiovascular system and brain tissue, resulting in thrombosis. This thrombosis is caused by 1) abnormalities in vascular endothelial cells, 2) excessive coagulation activity of blood coagulation factors (factor 4) Abnormal action of the dissolution system of the generated blood clot is known to be the cause. These causes act individually or in combination to cause excessive thrombus formation in blood vessels, and enzymes, factors related to thrombus formation, and substances that inhibit platelet aggregation are very useful preventive agents for various thrombotic diseases caused by excessive blood coagulation abnormalities. Or it can be used as a treatment.

For the prevention and treatment of thrombotic diseases, currently, 1) various anticoagulants such as heparin and coumarin, which inhibit the formation of blood clots by inhibiting the activity of blood coagulation factors, and 2) anticoagulants such as aspirin, which inhibit the formation of blood clots by inhibiting platelet aggregation. Platelet agents, and 3) thrombolytic agents such as urokinase, which dissolve already formed blood clots and inhibit the formation of blood clots, are being used. However, not only are they very expensive, but their use is limited due to bleeding side effects, gastrointestinal problems, and hypersensitivity reactions.

Recently, the development of antithrombotic agents has been intensively conducted from natural products derived from edible and medicinal crops that are safe and have antithrombotic activity, especially Ajoene from garlic, flavonoids from ginkgo leaves, rosemary essential oil, and Aes from chestnut trees. Aescin, etc.

Meanwhile, Aster scaber is a perennial plant belonging to the Asteraceae family. It mainly grows in mountains and fields and is distributed throughout Korea. Tuna has been reported to have neuroprotective effects, exhibit antioxidant activity, and is also known to prevent adipocyte differentiation in 3T3-L1 cells. At this time, platelets form aggregates when activated by agents such as collagen, adenosine diphosphate (ADP), and thrombin, which are secreted from activated vascular endothelium, which is commonly seen in the development of cardiovascular diseases. Once activated, platelets can bind to other platelets through integrin αIIbβ3, and this process also initiates the coagulation pathway. Therefore, platelets form aggregates and then form blood clots, causing blood vessel occlusion. This can lead to ischemia or even stroke, so it is important to inhibit platelet aggregation. However, even in these existing technologies, the anti-thrombotic and anti-platelet aggregation effects of Aster scaber have not been studied.

Accordingly, the present inventors completed the present invention by confirming the anti-thrombotic effect and anti-platelet aggregation effect of Aster scaber extract.

Jeon, S.-M., et al., Antioxidant activity of extracts and fractions from Aster scaber. Journal of the Korean Society of Food Science and Nutrition, 2012. 41(9): p. 1197-1204.

In order to solve the problems described above, the purpose of the present invention is to provide a pharmaceutical composition for preventing or treating thrombotic diseases containing Aster scaber extract as an active ingredient.

In addition, another object of the present invention is to provide a food composition for preventing or improving thrombotic diseases containing Aster scaber extract as an active ingredient.

Finally, another object of the present invention is to provide a health functional food composition for preventing or improving thrombotic diseases containing Aster scaber extract as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating thrombotic diseases containing Aster scaber as an active ingredient.

In one embodiment of the present invention, 'thrombotic disease' includes thrombosis, hypertension, stroke, cerebral infarction, angina, myocardial infarction, arteriosclerosis, peripheral artery occlusion, renal vein occlusion, central retinal vein occlusion, pulmonary thrombosis, and deep vein thrombosis. , portal vein thrombosis, cerebral venous sinus thrombosis, cerebral arteriosclerosis, heart disease, ischemic heart disease, intracranial hemorrhage, aneurysm, atherothrombosis, nephrosclerosis, and pulmonary embolism, but is not limited thereto.

In one embodiment of the present invention, ' Aster scaber extract' has platelet aggregation inhibitory activity, but is not limited thereto.

In one embodiment of the present invention, ' Aster scaber extract' has platelet granule secretion inhibitory activity, but is not limited thereto.

In addition, in order to achieve the above object, the present invention provides a food composition for preventing or improving thrombotic diseases containing Aster scaber as an active ingredient.

Finally, in order to achieve the above object, the present invention provides a health functional food composition for preventing or improving thrombotic diseases containing Aster scaber as an active ingredient.

Since the Aster scaber extract of the present invention effectively inhibits platelet aggregation activity and thrombus formation, it can be usefully used to prevent or treat various thrombotic diseases by acting as an effective inhibitor of thrombosis and platelet aggregation.

Figure 1 is a diagram showing the results of platelet aggregation by site of the Aster scaber extract of the present invention (platelet aggregation is induced by collagen (A), thrombin (B), and ADP (C)).
Figure 2 is a diagram showing the results of the Aster scaber extract of the present invention on inhibition of granule secretion from platelets at each site (ATP analysis (A) and Ca ²⁺ movement analysis (B)).
Figure 3 is a diagram showing the antiplatelet efficacy of the Aster scaber extract of the present invention against various extraction solvents (washed rat platelets were treated with Aster scaber extract using various solvents and then treated with collagen to induce platelet aggregation ).
Figure 4 is a diagram showing the antiplatelet activity of Aster scaber extract extracted using 70% ethanol of the present invention (platelet aggregation is induced by collagen (A), thrombin (B) and ADP (C); ATP analysis (D)).

The present invention provides a pharmaceutical composition for preventing or treating thrombotic diseases containing Aster scaber extract as an active ingredient.

The present invention provides a food composition for preventing or improving thrombotic diseases containing Aster scaber extract as an active ingredient.

The present invention provides a health functional food composition for preventing or improving thrombotic diseases containing Aster scaber extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

Before describing the present invention in detail, the terms used in the present invention are terms used to appropriately express the preferred embodiments of the present invention, which may vary depending on the intention of the user, operator, or customs in the field to which the present invention belongs. . Therefore, definitions of these terms should be made based on the content throughout this specification. Throughout the specification, when a part ‘includes’ a certain component, this means that it does not exclude other components but may further include other components, unless specifically stated to the contrary.

All technical terms used in the present invention, unless otherwise defined, are used with the same meaning as commonly understood by a person skilled in the art in the field related to the present invention. In addition, preferred methods and samples are described in this specification, but similar or equivalent methods are also included in the scope of the present invention. The contents of all publications incorporated by reference herein are hereby incorporated by reference.

In one aspect, the present invention relates to a pharmaceutical composition for preventing or treating thrombotic diseases containing Aster scaber extract as an active ingredient.

In addition, the Aster scaber extract may be extracted using water, ethanol, methanol, diethyl ether, ethyl acetate, acetone, or a mixture thereof as a solvent, preferably an ethanol extract, and more preferably It may be a 70% ethanol extract, but is not limited thereto.

The extract according to the present invention can be obtained by extraction and separation from nature using extraction and separation methods known in the art, and "extract" as defined in the present invention is obtained by extracting and separating Aster scaber using an appropriate solvent. It is extracted from, and includes, for example, crude extract, polar solvent-soluble extract, or non-polar solvent-soluble extract. The appropriate solvent for extracting the extract from Aster scaber may be any pharmaceutically acceptable organic solvent, and water or an organic solvent may be used, but is not limited thereto, for example, Alcohols with 1 to 4 carbon atoms, including purified water, methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, and benzene ), chloroform, ethyl acetate, methylene chloride, hexane, and cyclohexane can be used individually or in combination. As an extraction method, any one of the following methods can be selected and used: hot water extraction, cold needle extraction, reflux cooling extraction, solvent extraction, steam distillation, ultrasonic extraction, elution, and compression. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method.

There is no limitation to the method for producing the extract of the present invention, and any known method can be used. For example, the extract included in the composition of the present invention can be prepared in powder form from the primary extract extracted by the above-described hot water extraction or solvent extraction method by additional processes such as reduced pressure distillation and freeze-drying or spray drying. In addition, the primary extract was further purified into fractions using various chromatographies such as silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. You can also get it. Therefore, in the present invention, extract is a concept that includes all extracts, fractions, and purifications obtained at each stage of extraction, fractionation, or purification, as well as their dilutions, concentrates, or dried products.

The 'pharmaceutical composition' of the present invention may contain Aster scaber extract alone as an active ingredient, but may additionally contain pharmaceutically acceptable carriers, excipients, diluents, etc. depending on the formulation and purpose of use. there is.

In addition, the 'pharmaceutical composition' of the present invention may further include conventional fillers, weighting agents, binders, disintegrants, surfactants, anti-aggregants, lubricants, wetting agents, flavorings, emulsifiers, etc., and can be administered both parenterally and orally. You can use it. In addition, it can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions according to conventional methods. Suitable agents known in the art are preferably those disclosed in the literature (Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA). Carriers, excipients and diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline. These include cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, and mineral oil. When formulating the composition, it can be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations contain at least one excipient, such as starch, calcium carbonate, sucrose, lactose, It can be prepared by mixing gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, syrups, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. . It can be prepared as a preparation for parenteral administration, and its form can be a sterilized aqueous solution, non-aqueous solution, suspension, emulsion, freeze-dried preparation, suppository, etc. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogeratin, etc. can be used.

In the present invention, the dosage of the composition for preventing or treating thrombotic diseases can be appropriately selected by a person skilled in the art in consideration of the administration method, the age, gender and weight of the recipient, and the severity of the disease.

In the present invention, the pharmaceutical composition refers to the amount of the active ingredient or pharmaceutical composition that induces a biological or medical response in the tissue system, animal or human as considered by researchers, veterinarians, doctors or other clinicians, that is, alleviation of symptoms of the disease or disorder being treated. It can be administered in a therapeutically effective amount, which is an inducing amount. It is obvious to those skilled in the art that the therapeutically effective dosage and frequency of administration of the pharmaceutical composition of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by a person skilled in the art, and can be determined based on the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, the type of dosage form, and the patient's age, weight, and general health. It can be adjusted according to various factors, including condition, gender and diet, administration time, administration route and secretion rate of the composition, treatment period, and concurrently used drugs.

In the present invention, the pharmaceutical composition can be administered to an individual through various routes. All modes of administration are contemplated, for example, oral, rectal or by intravenous, intramuscular, subcutaneous, intrathecal or intracerebrovascular injection.

Additionally, the composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, or biological response regulators to prevent or treat thrombotic diseases or complications.

The term ‘prevention’ used in the present invention includes, but is not limited to, all acts of suppressing or delaying disease using the pharmaceutical composition. Accordingly, the present invention refers to both curative treatment and prophylactic aspects, and those that need to be treated include conditions that already have the disease as well as conditions in which the disease is to be prevented. In addition, it may refer to any act of administering the pharmaceutical composition of the present invention to an individual to improve disease symptoms or benefit the subject.

As used herein, the term 'treatment' refers to an approach to achieve a beneficial or desirable clinical result, whether or not the beneficial or desirable clinical result is detectable or possible, and whether partial or total, for the purposes of the present invention. It includes alleviation of symptoms, reduction of disease severity, stabilization (i.e., not getting worse) of disease, delay or reduction in disease progression, improvement or temporary relief and alleviation of disease condition without any effect.

In one aspect, the present invention provides a food composition for preventing or improving thrombotic disease containing Aster scaber extract as an active ingredient.

When the Aster scaber extract of the present invention is used as a food additive, it can be appropriately used according to conventional methods, such as adding the Aster scaber extract as is or mixing it with other foods or food ingredients. .

Additionally, the mixing amount of the active ingredient, Aster scaber extract, can be appropriately changed depending on the purpose of use (prevention, health, or therapeutic treatment).

There are no particular restrictions on the types of foods using the above active ingredients. Examples of foods to which the Aster scaber extract of the present invention can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, and various soups. , beverages, tea, drinks, alcoholic beverages, vitamin complexes, etc., and includes all health foods in the conventional sense.

When the food composition of the present invention is manufactured into a beverage, it may contain additional ingredients such as various flavoring agents or natural carbohydrates, like conventional beverages. The natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Natural sweeteners such as dextrin and cyclodextrin or synthetic sweeteners such as saccharin and aspartame may be used.

In addition to the above, the food composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, and alcohol. , carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may include pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages. These ingredients can be used independently or in combination.

In one aspect, a health functional food composition for preventing or improving thrombotic diseases containing Aster scaber extract as an active ingredient is provided.

The health functional food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc.

In the present invention, 'health functional food composition' refers to a food manufactured and processed using raw materials or ingredients with functionality useful to the human body in accordance with Act No. 6727 on Health Functional Food, and has It means taking it for the purpose of controlling nutrients or obtaining useful health effects such as physiological effects.

The health functional food composition of the present invention may contain common food additives, and its suitability as a food additive is determined according to the general provisions and general test methods of the food additive code approved by the Food and Drug Administration, unless otherwise specified. Determination is made according to relevant standards and standards. Items listed in the 'Food Additive Code' include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as dark pigment, licorice extract, crystalline cellulose, high-quality pigment, and guar gum; Examples include mixed preparations such as sodium L-glutamate preparations, noodle additive alkaline preparations, preservative preparations, and tar coloring preparations. For example, the health functional food in the form of a tablet is made by granulating a mixture of the active ingredient of the present invention with excipients, binders, disintegrants and other additives in a conventional manner, and then adding a lubricant and compression molding, or The mixture can be directly compression molded. In addition, the health functional food in the form of tablets may contain flavoring agents, etc., if necessary. Among capsule-type health functional foods, hard capsules can be manufactured by filling a regular hard capsule with a mixture of the active ingredient of the present invention mixed with additives such as excipients, and soft capsules can be prepared by mixing the active ingredient of the present invention with additives such as excipients. It can be manufactured by filling the mixture with a capsule base such as gelatin. The soft capsule may contain plasticizers such as glycerin or sorbitol, colorants, preservatives, etc., if necessary. The health functional food in the form of a pill can be prepared by molding a mixture of the active ingredient of the present invention and excipients, binders, disintegrants, etc., using a known method. If necessary, it can be coated with white sugar or other coating agents. Alternatively, the surface can be coated with substances such as starch or talc. Health functional food in the form of granules can be manufactured into granules by mixing a mixture of excipients, binders, disintegrants, etc. of the active ingredients of the present invention by a known method, and may contain flavoring agents, flavoring agents, etc., if necessary. You can.

Hereinafter, the present invention will be described in more detail through preparation examples and examples. However, the preparation examples and examples are presented as examples of the present invention, and if it is judged that a detailed description of a technology or configuration well known to those skilled in the art may unnecessarily obscure the gist of the present invention, the detailed description will be omitted. may be possible, and the present invention is not limited thereby. The present invention is capable of various modifications and applications within the description of the claims described below and the scope of equivalents interpreted therefrom.

<Preparation Example 1> Preparation of chemicals and reagents

Collagen and thrombin were purchased from Chrono-log (Havertown, PA, USA). The ATP assay kit was purchased from Biomedical Research Service Center (Buffalo, NY, USA).

<Preparation Example 2> Chamchae ( Aster scaber ) Extract preparation

Aster scaber was purchased from the Korea Plant Bank (Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea), and extracts for each part were prepared from the entire tuna, flowers, aerial parts, and roots and used in the experiment. Aster scaber was ground for each part and the extract was extracted through rotary evaporation using different solvents (50% ethanol, 70% ethanol, 100% ethanol or water) for 2 hours (Buchi, Flawil, Switzerland). Concentrated. The concentrated extract was frozen and lyophilized to obtain a powder that was used in the experiment.

<Preparation Example 3> Preparation of rat platelets

Seven-week-old Sprague-Dawley rats were housed in a pathogen-free environment (23 ± 2°C, humidity 50 ± 10%, 12-h light/dark cycle) and acclimatized for 1 week before experiments.

Blood was collected from rats via cardiac puncture and immediately added to acid citrate dextrose (ACD) solution as an anticoagulant. It was then centrifuged at 1,000 rpm for 7 minutes. Platelet-rich plasma was carefully separated and centrifuged at 2,000 rpm for 7 minutes. Pelletized platelets were resuspended in Tyrode buffer (137 mM NaCl, 12mM NaHCO3, 5.5mM glucose, 2mM KCl, 1mM MgCl2, and 0.3mM NaHSO4, pH 7.4), and washed platelets were obtained. Animal testing was approved by the Kyungpook National University Animal Experiment Ethics Committee (KNU-2013-001).

<Preparation example 4> Statistical analysis

The statistical significance of the data was analyzed by one-way analysis of variance (ANOVA) and measured using Dunnett’s post hoc test (SAS Institute Inc., Cary, NC, USA). All data were expressed as mean ± standard deviation (SD). A P value of 0.05 or less was considered statistically significant.

<Example 1> Chamchae ( Aster scaber ) Antiplatelet activity of extracts by region

1-1. Tuna ( Aster scaber ) Evaluation of platelet aggregation by region of extract

Platelet aggregation of Aster scaber extract was evaluated using a platelet aggregation meter (Chrono-log, Havertown, PA, USA). Washed platelets were incubated with or without Aster scaber extract for 2 minutes at 37°C in the presence of 1 mM CaCl ₂ . Cultures were then stimulated with collagen, thrombin, or ADP and incubated for an additional 5 min with continuous agitation.

Whole plants, flowers, aerial parts and roots of Aster scaber were prepared separately according to Preparation Example 2 above. Each tuna extract was made to 200 μg/mL and its antiplatelet activity was compared. As a result, the antiplatelet activity of the aerial part extract of Aster scaber was the least potent against all agents, while the extracts of the whole plant, flowers and roots of Aster scaber were significantly effective against collagen, thrombin and ADP. There was (see Figure 1).

1-2. Tuna ( Aster scaber ) Evaluation of inhibition of granule secretion from activated platelets in extracts from each region

The ATP activity analysis method is as follows. The washed platelets first underwent platelet aggregation. Platelets were incubated at 37°C for 2 minutes in the presence of 1 mM CaCl ₂ with or without Aster scaber extract. Afterwards, the cultures were stimulated with collagen (2.5 μg/mL) and continuously incubated for 5 min with continuous agitation. Upon completion of the reaction, the platelets were placed on ice to immediately quench the reaction and the supernatant was collected. Platelets were centrifuged, supernatants were collected, and ATP activity was measured using an ATP assay kit according to the manufacturer's protocol (Biomedical Research Service Center, State University of New York at Buffalo).

Additionally, [Ca ²⁺ ] _i movement analysis is as follows. Platelet-rich plasma was pre-incubated with Fura-2AM for 1 h at 37°C to obtain washed platelets, treated with 1 M CaCl ₂ with or without treatment of Aster scaber extract, and then incubated with collagen under continuous agitation. stimulated. [Ca ²⁺ ] _i was measured through the fluorescence intensity of Fura-2AM using a UV spectrophotometer.

Whole plants, flowers, aerial parts and roots of Aster scaber were prepared separately according to Preparation Example 2 above. Each Chamchae extract was made to 200 μg/mL and its granule secretion inhibitory activity in activated platelets was compared. As a result, whole plants, flowers and roots of Aster scaber showed a significant decrease in both calcium ion mobilization and ATP secretion. However, no significant decrease was seen in platelets treated with the aerial part extract of T. aquila. These results were identical to the results of the antiplatelet analysis above. In comparison, the flower extract of Aster scaber was confirmed to be the most powerful among all extracts (Figures 2A and B).

<Example 2> Tuna using various solvents ( Aster scaber ) Antiplatelet activity of extract

2-1. Cham fishing using various solvents ( Aster scaber ) Evaluation of platelet aggregation of extracts

Platelet aggregation of Aster scaber extract was evaluated using a platelet aggregation meter (Chrono-log, Havertown, PA, USA). Washed platelets were incubated with or without Aster scaber extract for 2 minutes at 37°C in the presence of 1 mM CaCl ₂ . Cultures were then stimulated with collagen, thrombin, or ADP and incubated for an additional 5 min with continuous agitation.

Extracts prepared using various solvents for the whole plant of Aster scaber were prepared separately according to Preparation Example 2 above. Each Chamchae extract was prepared at 200 μg/mL, 400 μg/mL, and 800 μg/mL, and the antiplatelet activity was compared. As a result, the 70% EtOH extract was confirmed to have the strongest inhibitory activity on platelet aggregation in a dose-dependent manner compared to the water extract, 50% ethanol extract, and 100% ethanol extract (Figure 3). Therefore, it was confirmed that the 70% ethanol extract had the strongest antiplatelet activity among all solvents.

2-2. Tuna ( Aster scaber ) Evaluation of platelet aggregation in 70% ethanol extract

Platelet aggregation was evaluated in the 70% ethanol extract of As ter scaber using a platelet aggregation meter (Chrono-log, Havertown, PA, USA). Washed platelets were incubated with or without Aster scaber 70% ethanol extract for 2 minutes at 37°C in the presence of 1 mM CaCl ₂ . Cultures were then stimulated with collagen, thrombin, or ADP and incubated for an additional 5 min with continuous agitation.

ATP activity analysis was performed as follows. The washed platelets first underwent platelet aggregation. Platelets were incubated at 37°C for 2 minutes in the presence of 1 mM CaCl ₂ with or without Aster scaber extract. Afterwards, the cultures were stimulated with collagen (2.5 μg/mL) and incubated continuously for 5 min with constant agitation. Upon completion of the reaction, the platelets were placed on ice to immediately quench the reaction and the supernatant was collected. Platelets were centrifuged, supernatants were collected, and ATP activity was measured using an ATP assay kit according to the manufacturer's protocol (Biomedical Research Service Center, State University of New York at Buffalo).

The whole plant of Aster scaber was extracted using 70% ethanol. In addition, 70% ethanol extracts of tuna tuna were prepared at 100 μg/mL, 200 μg/mL, and 400 μg/mL, and their antiplatelet activity was compared. As a result, it was confirmed that the 70% ethanol extract of the whole Aster scaber plant inhibited platelet aggregation induced by various agents in a dose-dependent manner (Figures 4A to C). Additionally, in the ATP analysis, it was confirmed that 70% ethanol extract of the whole plant of Aster scaber inhibited collagen-induced granule secretion from platelets in a dose-dependent manner (Figure 4D).

Through the above examples, it was confirmed that various parts of Aster scaber exhibit various antiplatelet activities. Therefore, the present inventor's goal was to identify the most potent part of Aster scaber and a solvent that extracts the most potent bioactive compounds that inhibit platelet aggregation. As a result, it was confirmed that the entire plant, including flowers, aerial parts, and roots, generally exhibits strong antiplatelet activity, and it was also inferred that the flowers and roots of Aster scaber also contain powerful bioactive compounds. In addition, in an antiplatelet activity experiment using various solvents of Aster scaber extract, it was confirmed that the extract using 70% ethanol exhibited the strongest antiplatelet activity compared to water, 50% ethanol, and 100% ethanol extracts, thereby confirming the present invention. Completed.

Claims (6)

A pharmaceutical composition for preventing or treating thrombosis containing Aster scaber flower extract as an active ingredient,
The composition wherein the thrombosis is caused by activated platelet aggregation and increased granule secretion of platelets. delete According to paragraph 1,
The composition is a pharmaceutical composition for preventing or treating thrombosis, which has platelet aggregation inhibitory activity. According to paragraph 1,
The composition is a pharmaceutical composition for preventing or treating thrombosis, which has the activity of inhibiting platelet granule secretion. A food composition for preventing or improving thrombosis containing Aster scaber flower extract as an active ingredient,
The composition wherein the thrombosis is caused by activated platelet aggregation and increased granule secretion of platelets. A health functional food composition for preventing or improving thrombosis containing Aster scaber flower extract as an active ingredient,
The composition wherein the thrombosis is caused by activated platelet aggregation and increased granule secretion of platelets.