JP5722052B2 - Thrombotic disease prevention food - Google Patents

Description

  The present invention relates to a thrombotic disease preventive food that can be easily ingested as a health supplement, and more particularly to a thrombotic disease preventive food containing a fermented product of a specific microorganism as an active ingredient.

  Looking at the medical care costs by injury and illness classification announced by the Ministry of Health, Labor and Welfare, the medical care costs for cardiovascular diseases consisting of hypertensive diseases, ischemic diseases and cerebrovascular diseases are the largest, accounting for 21.2% of the total medical costs is occupying. All of these diseases involve abnormal formation of blood clots in blood vessels.

  Blood has two actions: a coagulation system and a fibrinolytic system. When the blood vessel wall is damaged, platelets aggregate and primary hemostasis occurs, and then thrombin, a coagulation factor, acts on blood fibrinogen to form fibrin and complete hemostasis. On the other hand, tissue plasminogen activator (tPA), which is a fibrinolytic factor secreted from vascular endothelial cells, is converted into plasmin. Converts and the plasmin degrades fibrin. Failure of the balance between coagulation and fibrinolytic systems is known to cause thrombotic diseases such as cerebral infarction and myocardial infarction, and the development of new fibrinolytic substances greatly contributes to the prevention and treatment of these diseases It is considered possible.

  The present inventors have obtained a patent right (Patent Document 1) for a production method relating to a substance in which an enzyme and a trace component produced by culturing natto fungi are reversibly reduced to a stable state.

Japanese Patent No. 3902015

  An object of the present invention is to provide a functional food that prevents thrombotic diseases by enhancing tPA activity and inducing release of tPA from vascular endothelial cells.

  Under such problems, the present inventors added a low molecular peptide component reversibly reduced to a stable state to enzymes and trace components produced by Bacillus natto and added them to vascular endothelial cultured cells. The inventors found that tPA activity was enhanced and that when mice were orally ingested, blood tPA activity was enhanced, and the present invention was completed.

That is, the present invention
[1] A saccharification product obtained by hydrolyzing starch with amylase is used as a base material for a medium, and a yeast extract is added thereto to prepare a fermentation medium. Bacillus subtilis AK (accession number: FERM), which is a natto bacterium, is added to this medium. Inoculated P-18291), fermented and matured, thrombotic disease preventive food containing as an active ingredient a component obtained by separating the liquid component produced;
[2] The thrombotic disease-preventing food according to [1], wherein the fermentation is performed at pH 4.5 to 6.5 and 28 to 32 ° C. for 2 months or more;
[3] The food for preventing thrombotic disease according to the above [1] or [2], wherein the aging is aging performed for 4 months or more under conditions of pH 4.0 to 6.0 and 13 to 17 ° C;
[4] Thrombotic diseases include deep vein thrombosis, portal vein thrombosis, renal vein thrombosis, jugular vein thrombosis, Bad Chiari syndrome, axillary-subclavian vein thrombosis, cerebral venous sinus thrombosis and pulmonary thromboembolism One or more venous thrombosis selected from the group consisting of cerebral illness, or cerebral infarction, myocardial infarction, mesenteric artery thrombosis, acute limb thrombosis, hepatic artery thrombosis, renal artery thrombosis, splenic artery thrombosis The food for preventing thrombotic disease according to any one of [1] to [3], which is one or more arterial thrombosis selected from the group consisting of cerebral disease and obstructive arteriosclerosis; and [5] active ingredient The thrombotic disease-preventing food according to any one of [1] to [4], comprising 0.05 to 100% by weight based on the weight of the whole food.

  According to the present invention, it is possible to provide a food for preventing thrombotic diseases that can be easily orally ingested in daily life.

It is a graph which shows the molecular weight distribution of the active ingredient of this invention. 3 is a graph showing the effect of enhancing tPA activity by the active ingredient of the present invention, measured using a synthetic substrate S-2251 having a plasmin-specific cleavage site. It is the photograph (A) and graph (B) which show the tPA activity enhancement effect by the active ingredient of this invention measured using the fibrin plate. FIG. 2 is a photograph (A) showing tPA activity in a culture solution and a graph (B) showing the gene expression level of intracellular tPA measured using mouse brain vascular endothelium-derived cells bEnd.3. It is a graph which shows the binding property of the active ingredient of this invention with respect to tPA measured using the biomolecule interaction analyzer (IASYS). 2 is a photograph and a graph showing tPA activity in blood obtained by ingesting an active ingredient of the present invention into a mouse and collecting blood after 2 hours. 1 is a photograph and graph showing the effect of enhancing tPA activity in mouse blood when the mouse is ingested with the active ingredient of the present invention. It is the photograph which showed the enhancement effect of the tPA and uPA activity in the blood which blood ingested the lactic-acid-bacteria fermented extract to the mouse | mouth, and was collected 2 hours after that. It is the photograph which showed the enhancement effect of tPA and uPA activity in the blood which the mouse | mouth took the fermented natto fermented extract and was collected 2 hours after that.

  As an active ingredient contained in the food of the present invention, a saccharified product obtained by hydrolyzing starch containing corn starch with amylase is used as a base material for a medium, and a yeast extract is added thereto as a nitrogen source to prepare a fermentation medium. This medium is a component obtained by inoculating Bacillus subtilis AK (accession number: FERM P-18291), which is Bacillus natto, fermenting and aging, and then separating the produced liquid component.

  Here, the saccharified product used as the base material for the fermentation medium can be corn starch hydrolyzed with amylase separated and purified from corn grain, but crude corn starch is used instead of the purified corn starch. You can also. In addition to corn starch, soy flour, rice bran, or a mixture of these can be used as the substrate for the medium.

  Yeast extract used as a nitrogen source for fermentation media is generally added to bacterial cultures as a nitrogen source, such as dried water-soluble components extracted from brewer's yeast (Saccharomyces cerevisiae Meyen). Can be used.

  In addition to the above-described saccharified product and yeast extract, the fermentation medium used for the production of the active ingredient of the food of the present invention can contain organic substances such as proteins, inorganic salts, and the like as necessary. Examples of organic substances include soybean protein and other vegetable proteins, and examples of inorganic salts include calcium chloride, sodium chloride, and sodium phosphate.

  Further, it is also preferable to add sugars to the saccharified product obtained by hydrolyzing starch as a medium substrate with amylase. Examples of these sugars include sucrose (granulated sugar), glucose (dextrose), and starch syrup. Etc.

  The fermenting bacterium inoculated into the fermentation medium prepared as described above is Bacillus subtilis AK, which is Bacillus subtilis AK. ) FERM P-18291 ”. In addition, if desired, Lactobacillus lactobacilli (Lactobacillus), Lactococcus streptococcus (Streptococcus), yeast (Saccharomyces cerevisiae) and Aspergillus oryzae cells and fungi, which do not inhibit the growth of Bacillus subtilis AK An extract or a fungal fermentation extract can be inoculated in addition to Bacillus subtilis AK. Furthermore, the fermentation medium used for the production of the active ingredient of the food of the present invention, if desired, does not inhibit the growth of Bacillus subtilis AK, plant extracts such as Chinese cabbage, cabbage, carrot, medicinal carrot, parsley, celery, onion Can be added.

  Bacillus subtilis AK is an ordinary natto bacterium, Bacillus subtilis, UV, X-ray irradiation, high / low temperature environment (100 ° C, 0 ° C), competition with lactic acid bacteria, spore-forming medium [meat extract 5.0 wt. Subculture of resistant bacteria found under various conditions such as%, peptone 10.0%, sodium chloride 5.0%, agar 15.0%, vegetables (cabbage, carrot, celery, parsley) 65.0% by weight] Is obtained by repeatedly selecting.

The Bacillus subtilis AK strain thus obtained has the following mycological properties.
(A) Morphological properties 1 Cell shape and size Neisseria gonorrhoeae 1.0-1.2 × 3.0-50μm
2 No presence or absence of cell polymorphism 3 Presence or absence of motility (periflagellate flagella)
4 With or without spores Ellipse Almost center of cell (b) Culture characteristics 1 Meat agar plate culture circular colony White turbidity 2 Mice broth liquid culture upper or lower fungal aggregate (c) Biochemical properties 1 Gram staining Positive 2 Nitrate reduction Positive 3 MR test Negative 4 VP test Positive 5 Indole production Negative 6 Hydrogen sulfide production Negative 7 Use of citric acid Positive 8 Catalase positive 9 Range of growth
pH 5.5-7.0
Temperature 25 ℃ ~ 40 ℃

  The Bacillus subtilis AK strain is fermented for two months or longer on the above-mentioned fermentation medium with a pH of about 4.5 to 6.5 and a temperature of about 28 to 32 ° C. together with other fermenting bacteria including lactic acid bacteria if desired. In the pH range and temperature range lower than these ranges, it is estimated that the predetermined Bacillus natto used in this invention does not efficiently assimilate the sugar and nitrogen sources even when fermented for 2 months or more. The desired effect cannot be obtained sufficiently. On the other hand, in a pH range or temperature range higher than the above range, the desired effect of the tPA release-enhancing substance can be sufficiently obtained because the predetermined Bacillus natto does not efficiently assimilate the sugar and nitrogen sources. Absent.

  Following the fermentation, the fungi are aged on the same medium at a pH of about 4.0 to 6.0 and a condition of about 13 to 17 ° C. for 4 months or longer. In the pH and temperature ranges lower than these ranges, amino acids, lipoproteins, lipopolysaccharides (lipopolysaccharides), lipids, etc. that have various activities as fermentation products are sufficiently low in molecular weight even after aging for 4 months or longer. The expected effect of the obtained tPA release enhancing substance is not sufficiently obtained. On the other hand, in the pH range and temperature range higher than the above range, the activity is estimated to decrease, and the obtained tPA release-enhancing substance cannot sufficiently obtain the expected effect as described above.

In order to fractionate the liquid components produced through the fermentation and ripening stages, well-known separation means such as filtration or centrifugation can be employed. The fractionated food stock solution can be used as it is or after being concentrated or diluted. It can be used as an active ingredient of the food for preventing thrombotic diseases of the invention.
As an active ingredient of the food for preventing thrombotic diseases of the present invention, for example, an enzymine stock solution (ENM) manufactured by Enzamin Laboratories Co., Ltd. and an enzamin concentrate (ENM-HL) which is a 20-fold concentrated extract thereof are formulated. Can do.

  Incidentally, the components of the active ingredient thus obtained include substances for facilitating in vivo enzyme synthesis, that is, fragments obtained by reversibly cleaving the enzyme obtained by fermentation into active amino acid residues. In addition, it contains useful substances that can be used in vivo, such as adenine, guanine, cytosine, thymine, and uracil. Such useful substances include the anti-viral tissue active factor advocated by Besredka, the life-source stimulant described by Filatov, and combine these by biochemical reactions to act safely and effectively. It is considered that the culture filtrate was treated as described above.

  The food for preventing thrombotic diseases according to the present invention containing such an active ingredient stimulates a life phenomenon called fibrinolysis by blood, and appropriately controls the balance between the coagulation system and the fibrinolytic system to prevent the development of thrombotic diseases. To prevent.

  The thrombotic disease that can be prevented by the food for preventing thrombotic disease of the present invention is not particularly limited as long as it is a disease caused by a thrombus. For example, deep vein thrombosis, portal vein thrombosis, renal vein Thrombosis, jugular vein thrombosis, Bad Chiari syndrome, axillary-subclavian vein thrombosis, venous thrombosis selected from the group consisting of cerebral venous sinus thrombosis and pulmonary thromboembolism, or cerebral infarction, myocardial infarction, intestine Arterial thrombosis selected from the group consisting of mesenteric arterial thrombosis, lower limb acute arterial thrombosis, hepatic arterial thrombosis, renal arterial thrombosis, splenic arterial thrombosis, and obstructive arteriosclerosis.

  The food for preventing thrombotic diseases of the present invention comprises the active ingredient prepared as described above and excipients conventionally used in the food field (for example, starch or dextrin, cellulose, lactose, maltose, reduced lactose, reduced Maltose, sorbitol, mannitol, erythritol, xylitol, etc.) and adjuvants (eg, solvents, dispersion media, coatings, stabilizers, diluents, preservatives, preservatives, fungicides, antifungal reagents, osmotic reagents, absorption Inhibitors, disintegrants, emulsifiers, binders, lubricants, pigments, etc.) and mixed with pharmaceutical methods conventionally used in the food field, for example, tablets, capsules, granules, powders, extracts, solutions , Syrups, suspensions, emulsions.

  The food for preventing thrombotic diseases of the present invention is about 0.05 to 100% by weight, preferably about 0.1 to 90% by weight, based on the weight of the whole food, in terms of the liquid component produced by bacteria as an active ingredient. More preferably about 1 to 85% by weight, still more preferably about 5 to 80% by weight, and most preferably about 10 to 50% by weight.

  In another aspect, the present invention provides a saccharified product obtained by hydrolyzing starch containing corn starch with amylase for producing the thrombotic disease-preventing food as described above, and a yeast extract as a nitrogen source. To prepare a fermentation medium, inoculated with Bacillus subtilis AK (accession number: FERM P-18291), fermented and matured, and fractionated liquid components produced The present invention also relates to a method for preventing or treating a thrombotic disease characterized by the use of an ingredient, the use of Bacillus subtilis AK to produce a food for preventing a thrombotic disease, and the intake of a food for preventing a thrombotic disease.

  Hereinafter, the present invention will be described in more detail based on examples, but the present invention is not limited to these examples.

  Purified water 50 kg was added to yellow corn starch 2.3 kg, soybean peptone 0.5 kg, rice bran 0.5 kg, calcium chloride 80 g, and sodium chloride 150 g, and dissolved by heating. Next, this was cooled, and 50 g of amylase was added to allow sufficient saccharification. After saccharification is completed, 1.5 kg of granulated sugar, 1.5 kg of glucose (glucose), 450 g of yeast extract (Nippon Pharmaceutical Co., Ltd.), 1.5 kg of rice bran, 80 g of sodium phosphate, pressed vegetable juice (cabbage, carrot, celery, parsley) 5 kg and purified water were added to make the total amount 150 kg.

  And sodium hydroxide was added, pH was adjusted in the range of 7.2-7.6, this was put into the culture can and autoclaved at 120 degreeC for 20 minute (s). After cooling this, inoculate with Bacillus subtilis strain AK, ferment for 60 days at pH 4.5-6.5 in a constant temperature room of 30 ± 2 ° C., then pH 4.0-6.0 in a constant temperature room of 15 ± 2 ° C. The mixture was aged for 120 days under the following conditions. The supernatant was sterilized at 110 ° C. for 20 minutes and allowed to cool naturally to clarify the culture solution. This was filtered through a paper filter, adjusted to pH 3.5-3.7 with citric acid, sterilized at 95 ° C., and packed in a 5 gallon can at a temperature of 90 ° C. or higher. In this way, 125 liters of liquid food stock solution (ENM) was obtained.

The results of general analysis in 100 g of the resulting food stock solution (ENM) are shown in Table 1 below.
In addition, a liquid high-speed chromatogram (manufactured by Shodex: GPC) using the Tosoh column (TSKgel G2500PWXL) and a mobile phase of 55: 45: 0.1 mixture of water, acetonitrile and trifluoroacetic acid for the above food stock solution. Figure 21 shows the molecular weight distribution obtained by measuring size exclusion chromatography (SEC) with SYSTEM-21) and comparing the sensitivity of the detector (UV spectrophotometer: mV) with the elution time of a standard product with a known molecular weight. Table 1 shows the ratio (percentage) of the area of the molecular weight fraction in the figure to the whole.

Next, it was evaluated whether the low molecular weight substances contained in the obtained food stock solution (ENM) were denatured or decomposed by heat.
The molecular weight distribution of ENM heated at 121 ° C. for 30 minutes was measured by size exclusion chromatography using a TSK gel G2500PWXL column (manufactured by Tosoh Corporation). In addition, the molecular weight distribution of the same lot of ENM not exposed to high temperature (control ENM) was measured in the same manner. The results are shown in Table 2.

  As a result of molecular weight distribution measurement, ENM was found to be resistant to heat because no changes were observed in the molecular weight distribution of low molecular composition even when exposed to high temperature conditions.

In addition, it was evaluated whether the low molecular weight substances contained in the obtained food stock solution (ENM) were subject to denaturation or decomposition when subjected to strong acid conditions.
ENM (pH 3.7) was stirred while maintaining at 37 ° C., and hydrochloric acid was added to adjust the pH to 1.2. After standing for 15 minutes, the molecular weight distribution of the sample returned to the original pH 3.7 with sodium hydroxide was measured by size exclusion chromatography using a TSK gel G2500PWXL column (manufactured by Tosoh Corporation). In addition, the molecular weight distribution of the same lot of ENM not exposed to strong acid (control ENM) was measured in the same manner. The results are shown in Table 3.

  As a result of the molecular weight distribution measurement, ENM is resistant to strong acids even when exposed to strong acid conditions, so ENM is resistant to strong acids. Have been shown to be resistant.

  The above results prove that ENM is a stable raw material against high temperature and strongly acidic conditions, and it is not affected by high temperature or acidic conditions even when food processing or ingestion, and various processing and use Turned out to be possible.

  Next, the prepared ENM is put into a concentrator and subjected to primary concentration at a temperature of 40-60 ° C at a temperature of 40 ° C or lower, sterilized at 90 ° C for 30 minutes, and again put in a concentrator at a temperature of 40 ° C or lower. Thus, ENM-HL adjusted to a concentration ratio of 20 times through secondary concentration at a vacuum degree of 20-60 cmHg was obtained.

Examination of the effect on the plasmin activity of ENM-HL using the synthetic substrate S-2251 Next, the present inventors used the tissue type plasminogen activator (tPA) of ENM-HL (manufactured by Enzamin Laboratories). In order to directly examine the effect on plasmin activity, a synthetic substrate S-2251 having a specific cleavage site by plasmin was used. ENM-HL was diluted serially with physiological saline (100-0.13% by volume) and used as a sample.
To 10 μl of this sample, 90 μl of tPA (10IU / m1), 20 μl of Glu-plasminogen (200 μg / ml) and 100 μl of S-2251 (1 mM) were added and reacted, and the absorbance at 450 nm was measured every 2.5 minutes for 2 hours. The degree of increase (ΔA450nm) was calculated and tPA activity was evaluated.
As a result, significant enhancement of tPA activity by ENM-HL was observed (Fig. 2).

Examination of Effect of ENM-HL on tPA Activity Using Fibrin Plate Next, the present inventors examined the effect of ENM-HL on tPA activity using the fibrin plate method. ENM-HL was serially diluted with physiological saline (100 to 0.35% by volume) and used as a sample. A sample obtained by adding 10 μl of tPA (25 IU / ml) to 10 μl of the sample was added to the fibrin plate and allowed to react for 24 hours to evaluate tPA activity.
As a result, the effect of enhancing tPA activity by ENM-HL was also confirmed in the fibrin plate method (FIG. 3).

Examination of Effect of ENM-HL on tPA Activity and Production in Vascular Endothelial Cells Next, the present inventors examined the effect of ENM-HL on tPA activity and tPA production ability in vascular endothelial cells. As a vascular endothelial cell, mouse brain vascular endothelium-derived cell bEnd.3 cell was used. BEnd.3 cells are seeded at 2 × 10 ⁵ cells / well in a 24-well plate, and after reaching confluence, ENM-HL is added to serum-free medium at a concentration of 0.001 to 0.1% by volume for 6 hours, 12 hours. Incubated for 24 hours. The tPA activity in the culture was evaluated by fibrin zymography.
In addition, RNA was extracted from bEnd.3 cells after 24 hours of culture to synthesize cDNA, and the expression level of tPA mRNA was examined by real-time PCR. As an endogenous control, the GAPDH mRNA expression level was measured and corrected.
By adding ENM-HL, the tPA activity in the culture supernatant was enhanced after 6 hours, 12 hours, and 24 hours of culture (FIG. 4A). In addition, no increase in tPA mRNA expression was observed with ENM-HL, suggesting that tPA production of vascular endothelial cells was not affected (FIG. 4B).

Examination of binding ability of ENM-HL to tPA using biomolecular interaction analyzer (IASYS) The present inventors believe that ENM-HL contains a substance that acts directly on tPA, The binding property of ENM-HL to tPA was examined using an interaction analysis device (IASYS). tPA was immobilized on a cuvette and ENM-HL (1 to 20% by volume) was added to evaluate the binding.
As a result, strong binding to tPA was observed depending on the concentration of ENM-HL (Fig. 5). This suggests that ENM-HL contains a substance that strongly binds to tPA.

Examination of the effect of ENM-HL on tPA activity in the blood of mice Next, the present inventors examined C57BL6 / J mice in order to examine the effects of ENM-HL in vivo. Male C57BL6 / J mice (12 weeks old, body weight 25 ± 3g, 3 mice in each group) were orally administered 0.3 ml of ENM-HL (0.008-25% by volume), blood was collected 2 hours later, and plasma was collected. The euglobulin fraction was isolated. The fractions were evaluated for tPA activity by fibrin zymography.
As a result, enhanced tPA activity was observed at concentrations of ENM-HL 1% by volume and 0.1% by volume (FIG. 6).

Examination of the sustained effect of ENM-HL on tPA activity in mouse blood Next, in order to examine the sustained effect of tPA activity enhancing effect in vivo, 1% concentration of ENM-HL in which increased tPA activity was observed was changed to C57BL6. / J mice were orally administered, and blood was collected 1, 2, 3 and 4 hours later. Euglobulin fractions were isolated and evaluated for tPA activity by fibrin zymography.
As a result, it was confirmed that the enhancing effect of tPA activity was maintained for 4 hours or more, with the maximum value after 2 hours (FIG. 7).

  From the experimental results as described above, it was shown that ENM-HL enhances tPA activity in blood and the effect is maintained for a relatively long time.

Examination of the effect of medium-added components in mouse blood on tPA activity (1) Lactic acid bacteria fermented extract Next, the present inventors examined the effects of other fungal fermented extracts in vivo. Male C57BL6 / J mice (12 weeks old) were orally administered 300 μl of lactic acid bacteria fermentation extract (stock solution, 1-50% by volume) diluted with physiological saline, and euglobulin fraction was isolated from plasma collected 2 hours later . The tPA and uPA activities of the fractions were evaluated by fibrin zymography.
As a result, the effect of enhancing the activity of tPA and uPA was not observed in the lactic acid bacteria fermented extract (FIG. 8).
(2) Natto fermented extract Next, the present inventors examined the effects of other medium-added components in vivo. Male C57BL6 / J mice (12 weeks old) were orally administered 300 μl of Bacillus natto fermentation extract (20-fold concentrated solution, 0.2-25% by volume) diluted with physiological saline, and fractionated euglobulin from plasma collected 2 hours later. Was isolated. The tPA and uPA activities of the fractions were evaluated by fibrin zymography.
As a result, the effect of enhancing the activity of tPA and uPA was not observed in the Bacillus natto fermented extract (FIG. 9).

tPA is also used as a therapeutic agent for thrombotic diseases such as cerebral infarction, and its enhanced activity (activity enhancement) is considered to be effective for the treatment and prevention of these thrombotic diseases. This time, it was clarified that a component of a low molecular weight peptide derived from Bacillus natto enhances tPA activity in vivo. Since this ingredient has already been regarded as a health and nutritional food, it is expected to lead to the prevention of thrombotic diseases when taken as a food.
The food for preventing thrombotic diseases of the present invention belongs to the fields of nutritional functional foods, nutritional supplements, health supplements, foods for specified health use, particularly health supplements. The thrombotic disease-preventing food of the present invention comprises a microorganism culture that has been conventionally used for foods and has been confirmed to be safe, and can be easily ingested. The food for preventing thrombotic diseases of the present invention can effectively prevent the onset of thrombotic diseases by taking it easily and daily, and can reduce a great amount of medical costs related to thrombotic diseases.

Claims (5)

A saccharified product obtained by hydrolyzing starch with amylase is used as a base material for a medium, and a yeast extract is added thereto to prepare a fermentation medium. Bacillus subtilis AK (accession number: FERM P-18291), which is a natto bacterium, is added to this medium. ), Fermented and matured, and then a pharmaceutical composition for preventing thrombotic diseases comprising as an active ingredient a component obtained by separating the liquid component produced. The pharmaceutical composition for preventing thrombotic diseases according to claim 1, wherein the fermentation is fermentation performed at pH 4.5 to 6.5 and 28 to 32 ° C for 2 months or more. The pharmaceutical composition for preventing thrombotic diseases according to claim 1 or 2, wherein the aging is aging performed at pH 4.0 to 6.0 and 13 to 17 ° C for 4 months or longer. Thrombotic disease consists of deep vein thrombosis, portal vein thrombosis, renal vein thrombosis, jugular vein thrombosis, Bad Chiari syndrome, axillary-subclavian vein thrombosis, cerebral venous sinus thrombosis and pulmonary thromboembolism One or more venous thrombosis selected from the group, or cerebral infarction, myocardial infarction, mesenteric artery thrombosis, acute limb thrombosis, hepatic artery thrombosis, renal artery thrombosis, splenic artery thrombosis and occlusion The pharmaceutical composition for preventing thrombotic disease according to any one of claims 1 to 3, which is one or more arterial thrombosis selected from the group consisting of arteriosclerosis. The pharmaceutical composition for preventing thrombotic diseases according to any one of claims 1 to 4, comprising 0.05 to 100% by weight of the active ingredient based on the weight of the whole food.