KR20190088259A - Pharmaceutical composition comprising hexene fraction fractionated from ethanol extract of abeliophyllum distichum as an effective component for prevention or treatment of thrombosis and health functional food comprising the same - Google Patents

Abstract

The present invention relates to an antithrombotic composition comprising a hexene fraction of an Abeliophyllum distichum ethanol extract as an active ingredient, and more specifically, to a pharmaceutical composition and a health functional food for preventing or treating/alleviating thrombosis through blood coagulation inhibition comprising a hexene fraction of an ethanol extract prepared from leaves and stems of Abeliophyllum distichum as the active ingredient. As proved through an embodiment of the present specification, the hexene fraction of the Abeliophyllum distichum ethanol extract as the active ingredient of a pharmaceutical composition and a health functional food for prevention or treatment of thrombosis is expected to be used for the purpose of preventing and treating thrombosis such as ischemic stroke or hemorrhagic stroke through improvement of blood circulation since the hexene fraction of the Abeliophyllum distichum ethanol extract not only exhibits strong antithrombotic activity due to inhibition of thrombus formation-related enzymes and inhibition of blood coagulation factors, but also has excellent thermal stability and does not exhibit a loss in an inhibition effect on thrombus formation-related enzymes and blood coagulation factors even in an acidic condition of pH 2 and in blood plasma. Therefore, the present invention is very useful in pharmaceutical and food industries since the active ingredient is processed in various forms such as extract, powder, pill, tablet, and the like to be prepared in forms which can be taken at any time.

Description

TECHNICAL FIELD [0001] The present invention relates to a pharmaceutical composition for preventing or treating thrombosis and a health functional food containing the hexane fraction as an active ingredient. HEALTH FUNCTIONAL FOOD COMPRISING THE SAME}

The present invention relates to the use of Abeliophyllum distichum ethanol extract as an active ingredient, and more particularly to an antithrombotic composition comprising a hexane fraction of an ethanol extract prepared from the leaves and stems of a beech tree as an active ingredient, A pharmaceutical composition for preventing or treating / improving thrombosis, and a health functional food.

The blood as a constituent of human body has various important functions such as oxygen and nutrients, the functions of carrying and buffering of waste products, maintenance of body temperature, control of osmotic pressure and maintenance of ion balance, maintenance of moisture, regulation of fluidity, maintenance and regulation of blood pressure, have. Normal blood circulation facilitates blood circulation by complementary regulation of the blood coagulation system and thrombolysis system in the body. Among them, the mechanism of the blood coagulation system is that the platelets adhere to the blood vessel walls and coagulate to form platelet thrombus , The blood coagulation system is activated and fibrin thrombi are formed centering on platelet aggregation mass.

On the other hand, the production of fibrin thrombus is activated by thrombin involved in fibrin clotting through several steps of a number of blood coagulation factors to finally produce a fibrin monomer from fibrinogen. The fibrin monomers are polymerized by calcium, Cells to form a cross-linked fibrin polymer by factor XIII and produce a permanent thrombus. In addition, thrombin plays a pivotal role in thrombus formation by activating platelet, V factor, and Factor VII to promote blood coagulation. Therefore, the activity inhibitor of thrombin can be used as a prophylactic and therapeutic agent very useful for various thrombotic diseases caused by excessive blood coagulation. On the other hand, activation of prothrombin after sequential activation of factor XII, factor XI, factor XII, factor IX and factor X is finally known to activate thrombin in the endogenous thrombosis pathway, and the specific inhibition of blood clotting factor In the extrinsic thrombosis pathway, thrombogenesis is known to be caused by the activation of factor II (prothrombin), factor V, factor VII, and factor X. Up to now, various anticoagulants such as heparin, coumarin, aspirin, and europine have been used for the prevention and treatment of thrombotic diseases. However, these anticoagulants and thrombolytic agents are not only very expensive but also have hemorrhagic side effects, The use thereof is limited due to the reaction and the like.

On the other hand, the Abelophyllum distichum ) is a rare plant of the first genus in the world, and is a Korean specialty plant that is native to Korea only. The Ministry of Environment has designated wild tree as a protected wild plant and it is now native to Jincheon, Goesan, Yeongdong, Jeollabuk-do, Chungcheongbuk-do, and is a rare natural monument. It is a tree of about 1m in height and belongs to the ash tree. It is spread out in four directions, and it is turned purple and has a square shape. Leaves are opposite, ovate or ovate ovate, narrow at both ends, flat at edge and short stalk. The flowers bloom before the leaves and run on the stamina. The shape of the seeds is called fan-shaped, like a fan.

The present study was carried out to investigate the characteristics of the spruce tree and the characteristics of the growth and growth characteristics of the spruce tree (Park, Seonghyuk, 2016. Korea Master in Youngmyun University, "Characteristics of Vegetation Structure , "Plant Biotechnology" (414: 94-99), recent analysis of the components of leaf and stem of Pinus rigida and antioxidant and whitening of leaf and immature seed of anticancer, anti-inflammatory, Journal of life sciences 27: 536-544; 2011; The Journal of Herbology 26: 95-99), anti-wrinkle (Kim, Nam-young et al., 2015. Korean journal of medicinal crop science 23: 231-236). However, there is no known strong antithrombotic activity due to the inhibition of thrombogenesis-related enzymes and blood coagulation factors in the above-ground microbial trees.

In addition, most patents relating to the present invention include patent for cosmetics utilizing the fragrance of spruce tree and antioxidant and anti-inflammatory activity, and in Korean Patent No. 10-1729210 [ Product composition], registered trademark No. 10-1560672 [Deodorant composition containing vinegar extract], registered trademark 10-1181998 [cosmetic composition for removing odor of elderly people containing vinegar extract as an active ingredient] Patent No. 10-1470887 discloses a perfume composition which reproduces the fragrance of the scarlet tree, a fragrance composition recreating the fragrance of the scarlet flower, and a composition for external application for skin containing the fragrance composition recited in the registered patent 10-1773090 -1191225 discloses a cosmetic composition, and a registered trademark 10-0954695 discloses a cosmetic composition containing an extract of Aspergillus as an active ingredient. In 2007, No. 10-0656287 [an anti-inflammatory agent containing a herb extract], and in a patent No. 10-0706131 [an anti-cancer agent containing a herb extract]. In recent years, Korean Patent Laid-Open No. 10-2017-0122317 [Shampoo composition for preventing hair loss using waxy extract] and Japanese Patent Laid-Open No. 10-2017-0122317 [Shampoo composition for preventing hair loss using waxy extract] Patent No. 10-2017-0122315 [Cosmetic Composition for Skin Using Vine Trefoil Extract], Published Japanese Patent Application No. 10-2015-0068643 [Disclosed is a specific odor eliminating agent for livestock products and aquatic products using an extract of Alaska pollack and its preparation method] 10-2013-0074172 discloses a composition for cosmetics using a fermented product of a fine line tree. However, up to now, no scientific research or patent literature has been known about the potent thrombogenesis-related enzymes and antithrombotic effects of inhibiting blood coagulation factors in the extracts of Echinochloa crus-galli.

KR 10-1729210 B KR 10-2017-0122317 A

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide an antithrombotic composition comprising an extract of hexane will be.

In order to solve the above-mentioned problems, the present invention provides a pharmaceutical composition for preventing or treating thrombosis, which comprises an extract of hexane of an extract of Helicobacter pylori as an active ingredient.

It is preferable that the above-mentioned tree is a leaf or stem of a spike tree.

The present invention also provides a blood coagulation inhibitor containing the above-described effective ingredient.

The present invention also provides a health functional food for preventing or ameliorating thrombosis containing the above-mentioned effective ingredient.

The pharmaceutical composition for the prevention or treatment of thrombosis according to the present invention and the hexane fraction of the extract of barnyard grass ethanol as an active ingredient of the health functional food can be used for the treatment of thrombosis-related enzyme inhibition and blood coagulation factor , And exhibits excellent thermal stability and does not exhibit the loss of the thrombogenic enzyme and blood coagulation factor inhibitory effect in an acidic condition of pH 2 and in plasma, It is expected that the active ingredient can be used for prevention and treatment of thrombosis such as hemorrhagic stroke, and the effective ingredient can be prepared into various forms such as extract, powder, ring, tablet, etc., It is a very useful invention in the pharmaceutical industry and the food industry.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a photographic view of the above-mentioned tree top used in the embodiment of the present invention. FIG. Here, 1: ground part, 2: leaf, and 3: stem are respectively represented.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention prepared an ethanol extract of Zygomycota leaves and stems prepared by a certain method and then fractionated with hexane, ethyl acetate and butanol sequentially in organic solvent, Finally, a water residue after the fractionation was prepared. The extract and fractions were evaluated for their antithrombotic activity, and the hexane fraction of the extract of the spruce tree was recovered as an anti-thrombogenic component. The fraction had excellent thermal stability and acid stability To thereby make use of the above ingredients as a pharmaceutical composition for antithrombotic activity and as a health functional food.

In order to develop a pharmaceutical composition and a health functional food for prevention or treatment / improvement of thrombosis using spikelets which are known to have various physiological activities in the private sector, Each of the extracts was fractionated with hexane, ethyl acetate and butanol in order to prepare a water residue. The antimicrobial activity of the extracts was measured by thrombin time, thrombin time, The prothrombin time and the activated partial thromboplastin time (aPTT) were evaluated to show that the hexane fraction of the ethanol extract of Pinus rigida exhibited excellent antithrombotic activity following strong anticoagulant inhibition Respectively.

The extracts of 25.9 g and 5.8 g per 100 g of leaf and stem ethanol extracts of Leuconthus can be recovered, and the hexane fractions of leaf and stem ethanol extracts can recover 2.56 g and 1.19 g, respectively, It was expected to be possible. The ethanol extracts of Leuconostoc sp. Leaf and stem showed weak coagulase activity (thrombin and prothrombin) and blood coagulation factor inhibitory activity. However, the hexane fraction of Leuconostoc sp. Leaf extract showed 2.11 times, 1.25 And 1.92-fold increased thrombin time, prothrombin time, and apathy time, and the hexane fraction of the ethanol extract of barley trunks showed 1.36-fold, 1.27-fold, and 1.54-fold increased thrombin Time, prothrombin time, and apathy time, confirming the potent antithrombotic activity of these fractions. In addition, the hexane fractions of the ethanol extract of Echinosophora koreensis showed anticoagulant activity in a concentration dependent manner.

Accordingly, the present invention provides a pharmaceutical composition for the prevention or treatment of thrombosis, which comprises a hexane fraction of a plant extract of Hercules, USA as an active ingredient.

It is preferable that the fine line tree is a ground part of a fine line tree, for example, a leaf or a stem.

In addition, the present invention provides a blood coagulation inhibitor containing the hexane fraction of the above-mentioned bentwood ethanol extract. The above medicinal use is based on the strong hemostatic enzyme and blood coagulation factor inhibitory action of the hexane fraction of the waxy ethanol extract of the present invention.

The present invention also provides a health functional food for preventing or ameliorating thrombosis, which contains the hexane fraction of the above-mentioned extract of Helicobacter sp. As an active ingredient.

Hereinafter, the production method and efficacy test of the hexane fraction of the extract of the vermicelli ethanol of the present invention will be described in more detail.

The inventors of the present invention have found that, in order to achieve the object of the present invention, Preparing an extract of the above-ground wood part; Preparing an organic solvent fraction of hexane, ethyl acetate, butanol, and finally a water residue after fractionation; Experiments were conducted on the antithrombotic activity evaluation of the extract and fractions and the evaluation of the stability of the active substance.

The "hexane fraction of the extract of the non-waxy ethanol extract" contained in the composition of the present invention is prepared by separately preparing leaves, mm or less, and concentrating it under a reduced pressure, and preparing the hexane fraction by a sequential organic solvent fraction.

According to the embodiment of the present invention, the hexane fraction of the ethanol extract of Leucon leaf showed a prolonged thrombin time, prothrombin time and apathy time, which were 2.11 times, 1.25 times and 1.92 times longer than those of the no-added solution at the concentration of 5 mg / The hexane fraction of the stem ethanol extract showed 1.36 times, 1.27 times, 1.54 times longer thrombin time, prothrombin time, and apathy time at the concentration of 5 mg / ml, respectively, But still showed excellent antithrombotic activity. Considering that aspirin (product name: Protect), which is used as an anti-thrombotic agent in clinical practice, prolongs thrombin time, prothrombin time, and apathy time by 1.95 times, 1.40 times, and 1.63 times at 1.5 mg / ml concentration, Activity.

The hexane fraction of the herbal extract of the present invention may be prepared into powder by conventional powdering processes such as vacuum drying and freeze drying, spray drying and the like. They are not degraded by various degradation enzymes in the plasma, and maintain their activity even at 100 ° C heat treatment and pH 2 in human body.

The active ingredient of the present invention can be used for the prevention or treatment of various diseases associated with thrombosis. Such diseases include, for example, arterial thrombosis such as acute myocardial infarction, chest pain, dyspnea, loss of consciousness, ischemic stroke, hemorrhagic stroke, headache, dyskinesia, sensory abnormality, personality change, visual disturbance, epileptic seizure, , Deep vein thrombosis, lower limb edema, pain and acute peripheral artery occlusion. Vein thrombosis can include deep vein thrombosis, portal vein thrombosis, acute renal vein thrombosis, cerebral sinus thrombosis, and central retinal vein occlusion.

The pharmaceutical composition containing the active ingredient of the present invention may be formulated into tablets, capsules, suspensions, emulsions, oral preparations such as syrups and aerosols, injections of sterilized injection solutions And may be administered by various routes including oral administration or intravenous, intraperitoneal, subcutaneous, rectal, topical administration, and the like.

Such pharmaceutical compositions may further comprise carriers, excipients or diluents, and examples of suitable carriers, excipients or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, But are not limited to, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, amorphous cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, And the like. In addition, the pharmaceutical composition of the present invention may further include a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, an antiseptic, and the like.

In a preferred embodiment, the solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient, for example starch, calcium carbonate, Sucrose, lactose, gelatin and the like are mixed and formulated. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used.

Examples of the oral liquid preparation include suspensions, solutions, emulsions, syrups and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, Perfumes, preservatives, and the like.

As a preferable specific example, the preparation for parenteral administration includes sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-drying agents, suppositories, and the like. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Injectables may include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, preservatives, and the like.

The active ingredient of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, The sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

As a preferable example, the effective amount of the active ingredient in the pharmaceutical composition of the present invention may vary depending on the age, sex, and body weight of the patient, and generally 1 to 5,000 mg, preferably 100 to 3,000 mg per kg of body weight per day Or every other day or one to three times a day. However, the dosage may not be limited in any way because it may be increased or decreased depending on route of administration, severity of disease, sex, weight, age, and the like.

The pharmaceutical composition of the present invention can be administered to a subject through various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

In the present invention, "administration" means providing a predetermined substance to a patient by any suitable method, and the administration route of the pharmaceutical composition of the present invention is either oral or non-oral May be administered orally. The composition of the present invention may also be administered using any device capable of delivering an effective ingredient to a target cell.

In the present invention, the term "object" includes, but is not limited to, human, monkey, cow, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit or guinea pig , Preferably a mammal, more preferably a human.

In addition, the health functional food of the present invention can be variously used for foods and beverages effective for prevention or improvement of thrombosis. Examples of foods containing the active ingredient of the present invention include various foods, beverages, gums, tea, vitamin complex, health supplement foods and the like, and they can be used in the form of powder, granule, tablet, capsule or beverage .

The active ingredient of the present invention may generally be added in an amount of 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.

The health functional food of the present invention may contain, as an additional ingredient, a food-acceptable food-aid additive such as natural carbohydrates and various flavors, in addition to containing the above-mentioned compound as an essential ingredient in the indicated ratio.

Examples of the natural carbohydrate include sugar sugars such as glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol.

Examples of the flavoring agents include natural flavoring agents such as tautatin, rebaudioside A and stiglycerin such as glycyrrhizin, and synthetic flavoring agents such as saccharin and aspartame. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the health functional food of the present invention. In addition to the above, the health functional food of the present invention may contain various kinds of nutrients, vitamins, minerals, flavors such as synthetic flavors and natural flavors, colorants and heavy stabilizers, pectic acid and its salts, alginic acid and its salts, Thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like.

In addition, the health functional food of the present invention may contain flesh for producing natural fruit juice, fruit juice drink, vegetable drink and the like. These components may be used independently or in combination. The ratio of such additives is generally selected in the range of 0.01 to about 20 parts by weight per 100 parts by weight of the active ingredient of the present invention.

Hereinafter, specific methods of the present invention will be described in more detail by way of examples. The following examples are only a preferred embodiment of the present invention, and the scope of the present invention is not limited to the scope of the following examples.

[Example]

Example  1: Ethanol extracts of leaves and stems from axel Fraction  Analysis of ingredients and components of each sample

(Yuncheon Farm: Endangered Species of Wild Fauna and Flora: 2012-2), and the leaves were separated into leaves and stems and used for the production of ethanol extracts, respectively. Ethanol extracts were prepared by adding 10 times ethanol (95%, Duksan, Korea) to each sample and then extracting the extracts three times at room temperature. The extracts were collected by filtration and concentrated under reduced pressure to prepare powders. Subsequently, the ethanol extracts were fractionated with hexane, ethyl acetate and butanol, and finally the water residue was recovered. The extraction efficiency of ethanol extract, the organic solvent fraction efficiency, and the component analysis results of the extract / fraction are shown in Table 1. Total polyphenols, total flavonoids, total sugars and reducing sugar contents were measured by the compositional analysis of the prepared ethanol extracts and fractions. Total polyphenol content was determined by adding 50 μl of Folin-ciocalteau and 100 μl of saturated Na ₂ CO ₃ solution to 400 μl of the extract solution, leaving it at room temperature for 1 hour and measuring the absorbance at 725 nm. Tannic acid was used as a standard reagent. The total flavonoid content of each sample was measured by stirring for 18 hours in methanol. To the 400 μl of the filtered extract, 4 ml of 90% diethylene glycol was added, and 40 μl of 1N NaOH was added thereto, followed by reacting at 37 ° C. for 1 hour. As a standard reagent, rutin was used. Reducing sugar was quantified by DNS method and total sugar was quantified by phenol-sulfuric acid method.

As a result, as shown in Table 1, the extraction efficiency was 25.9% for the ethanol extract of Leucumber leaf and 5.8% for the ethanol extract of the stem, and the extraction efficiency was 4.5 times higher than that of the leaf stem. The ethyl acetate fractions (33.7%), the hexane fractions (9.9%) and the water residues (3.5%) were the most abundant in the leaf extract fractions. On the other hand, in the case of the stem extract, the butanol fraction (50.5% of the extract), the water residue (26.6%), the hexane fraction (20.6%) and the ethyl acetate fraction (12.2% The contents of total flavonoids and total flavonoids were higher in the leaf extracts than in the stem extracts. In the fractions, the leaves showed higher total flavonoids and total flavonoids in the ethyl acetate fraction and the stem fraction in the butanol fraction. On the other hand, in the case of total sugar and reducing sugar content, the leaves showed the highest content in the butanol fraction and the stem in the water residue. These results indicate that the leaves and stems of Zebranus are distinctly different components.

Example  2: Leaf and stem ethanolic extract and its object Fraction  Evaluation of blood coagulation inhibitory activity

The blood coagulation inhibitory activity (thrombogenesis inhibitory activity) of ethanol extracts and fractions thereof from the leaves and stems of Zygomycota prepared in Example 1 was evaluated and compared with the previously reported method (Sohn et al., 2004. Kor. Similar to Pharmacogn 35. 52-61; Kwon et al., 2004. J. Life Science, 14. 509-513; Li et al 2010. J. Life Science, 20. 922-928), thrombin time, prothrombin time, The apathy time was measured and evaluated. Plasma was obtained from commercial control plasma (MD Pacific Technology Co., Ltd., Huayuan Industrial Area, China), and thrombin time, prothrombin time and apitime time were measured by the following procedure.

Thrombin Time

50 μl of 0.5 U thrombin (Sigma Co., USA) and 50 μl of 20 mM CaCl ₂ and 10 μl of various concentrations of the sample extract were mixed in a tube of Amelung coagulometer KC-1A (Japan) for 2 minutes at 37 ° C., After the addition, the time until the plasma coagulated was measured. As a control, aspirin (Sigma Co., USA) was used and DMSO was used as a solvent control instead of the sample. DMSO showed a solidification time of 30.5 sec. The thrombin inhibitory effect was expressed as the average value of the experiments repeated three or more times. The thrombin inhibitory activity was expressed by the value obtained by dividing the solidification time at the time of addition of the sample by the solidification time of the solvent control.

Prothrombin time ( prothrombin  time)

Add 70 μl of standard plasma (MD Pacific Co., China) and 10 μl of various concentrations of sample solution to tubes of Amelung coagulometer KC-1A (Japan), heat at 37 ° C for 3 minutes, add 130 μl of PT reagent, The time from the start of the experiment to the start of the experiment was expressed as the average value of the experiments repeated three times. As a control, aspirin (Sigma Co., USA) was used and DMSO was used as a solvent control instead of the sample. DMSO showed a clotting time of 16.7 seconds. The prothrombin inhibitory activity was expressed as the value obtained by dividing the solidification time at the time of addition of the sample by the solidification time of the solvent control.

aPTT (activated Partial Thromboplastin  Time)

After adding 100 μl of plasma and 10 μl of various concentrations of sample extract to a tube of Amelung coagulometer KC-1A (Japan) and heating at 37 ° C for 3 minutes, 50 μl of aPTT reagent (Sigma, ALEXIN ^™ ) And cultured for 3 minutes. Then, 50 μl CaCl ₂ (35 mM) was added and the time until the plasma coagulated was measured. As the solvent control, DMSO was used instead of the sample. In this case, the solidification time was 58.1 seconds. The results of aPTT were expressed as the mean value of three repeated experiments. The activity of inhibiting blood coagulation factor was represented by aPTT divided by the aPTT of the solvent control when the sample was added.

As a result, as shown in Table 2, the ethanol extracts of the leaves and stems showed weak thrombin inhibition and blood coagulation factor inhibition at a concentration of 5 mg / ml. The thrombin time was 1.17 to 1.22 times and the apathy time was 1.19 to 1.38 The boat was extended. Among the various fractions, the hexane fraction and the ethyl acetate fraction showed better anticoagulant activity than the ethanol extract, but the butanol fraction and the water residue showed weaker activity than the ethanol extract. Therefore, concentration-dependent anticoagulant activities of hexane fraction and ethyl acetate fraction were measured, and the results are shown in Table 3.

As a result, the hexane fraction of each leaf and stem extract showed better activity as the concentration was increased. At the concentration of 7 mg / ml, thrombin time, prothrombin time, and apathy time were 2.54 ~ 3.47 times, 1.56 ~ 1.69 times and 1.63 ~ 3.09 times, respectively. Leaf extract showed better anticoagulant activity than stem extracts. At this time, aspirin used as a control was lengthened by 1.95 times, 1.40 times, and 1.63 times of thrombin time, prothrombin time, and apathy time at a concentration of 1.5 mg / ml. These results suggest that the hexane fraction of Euglena ethanol extract can be developed as a potent antithrombotic agent that can replace gastrointestinal aspirin.

Example  4: Ethyl alcohol extract of Leucocephalus Hexen Fraction  Plasma, acid and thermal stability assessment

The plasma stability, thermal stability and acid stability of the antioxidant activity of the hexane fraction of the ethanol extracts of the leaves and stems of Example 1 were confirmed. The extract maintained its excellent activity without heat treatment at 100 ° C for 1 hour, 1 hour treatment with pH 2 (0.01M HCl), and 1 hour treatment with plasma without loss of anticoagulant activity. These results suggest that ethanol extracts from the barnyardgrass can be practically used as antithrombotic agents, and it may be possible to supplement and replace aspirin with side effects such as gastrointestinal complaints.

Claims (4)

A pharmaceutical composition for the prevention or treatment of thrombosis, comprising an extract of hexane fraction of a plant extract of Echinochloa crus-galli as an active ingredient. 2. The pharmaceutical composition according to claim 1, wherein the fine line is a leaf or stem of a fine line tree. 4. A blood coagulation inhibitor comprising the effective ingredient according to any one of claims 1 to 3. A health functional food for preventing or improving thrombosis comprising the effective ingredient according to any one of claims 1 to 3.

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