JP2005179252A - Inhibitor of thrombogenesis - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an inhibitor of thrombogenesis, usable in wide food and drink; and to provide the food and drink containing the inhibitor. <P>SOLUTION: The inhibitor of the thrombogenesis contains Gymnema sylvestre R. Br. or an extract of the Gymnema sylvestre R. Br. Preferably, the extract of the Gymnema sylvestre R. Br. is the extract extracted by using one or more kinds selected from the group consisting of water, a base, an acid and alcohols. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a thrombus formation inhibitor containing Gymnema or an extract of Gymnema, and a food or drink containing the same.

A thrombus can be formed into an insoluble polymer together with platelets, leukocytes and the like and solidified on the inner wall of a blood vessel while a protein called fibrinogen is activated and converted into fibrin. When the body is normal, the fibrinolytic enzyme that works to dissolve fibrin, which is the source of the thrombus, prevents thrombus. However, when the fibrinolytic enzyme is insufficient, fibrin cannot be dissolved and a thrombus can be formed.
The formed thrombus is deposited in the blood vessels, reducing the cross-sectional area of the blood vessels and impeding blood circulation, so that the blood cannot normally supply nutrients and oxygen in cells and tissues, The waste of cells and tissues cannot be discharged, causing problems such as accumulation of toxic substances.
In blood vessels, the symptoms caused by blood clots called blood clots are called thrombosis in a broad sense (hereinafter simply referred to as “thrombosis” in the broad sense) and are caused by blood clots. The pathological condition is divided into thrombosis and embolism in a narrow sense. In a narrow sense, thrombosis is a symptom caused by partial or complete blockage of blood flow at the site where the thrombus is formed. It refers to a pathological condition caused by physical or complete occlusion.
Such thrombosis induces various diseases depending on the site of the blood vessel where the thrombus has occurred. In particular, serious symptoms such as stroke, cerebral hemorrhage, cerebral infarction, heart failure, myocardial infarction, heart palsy occur when it occurs in the cerebral blood vessel or cardiovascular, causing half-body involuntary and death in severe cases is there.

Currently, in order to solve thrombosis, research and development of antithrombotic agents and thrombus formation preventive agents that suppress the formation of thrombus and thrombolytic agents that dissolve the generated thrombus are mainly conducted.
As an antithrombotic agent or an agent for preventing thrombus formation, aspirin that inhibits blood coagulation by inhibiting the adhesion of platelets to the blood vessel wall, heparin (Heparin), coumarin (Coumarin) that blocks the body's intrinsic blood coagulation pathway ) Etc. are currently used in clinical practice. Recently, eicosapentaenoic acid (EPA), prostacycline (PG12) derivatives and the like have been commercialized. However, since these drugs have no specificity, they may affect parts other than the thrombus in the living body, and may cause bleeding or the like when remaining in the living body. In addition, antithrombotic activities such as hirudin, synthetic antithrombin, and ticlopidin have been reported, but have not yet been put into practical use.
As a thrombolytic agent, a plasminogen activator (Plasminogen activator) such as streptokinase or urokinase is intravenously injected into a patient in which a thrombus has been generated to activate the thrombolytic system in the body. Is commonly used. The effect of dissolving thrombus has been proved in many clinical experiments, but, like antithrombotic agents or antithrombotic agents, it has no specificity for thrombus and has side effects such as general bleeding during the treatment of thrombus. is there. Also, tissue-type plasminogen activator (tPA) is considered to be an ideal thrombolytic agent because of its high selectivity for thrombus, but there are some differences as a result of actual application to clinical treatment. However, there were side effects such as general bleeding. In addition, since the half-life in the blood is very short and the duration of the drug is short, the dose must be large in order to maintain the drug in the body, so the treatment cost is much higher than that of conventional thrombolytic agents. There is a problem that it is expensive.

Although these medicines are used to prevent the formation of blood clots, they do not show a significant effect on thrombus removal and may cause serious side effects. Research on ingredients or food ingredients having a function of preventing illness throughout life and regulating or activating constitution has also attracted attention.
As food ingredients, materials such as nattokinase, polyunsaturated fatty acids, glucosamine, and onion skin (for example, see Patent Document 1) are known, but there are problems with flavor and properties, and they are widely applied to foods. could not.
In addition, recently, a patent for a kiwifruit extract (for example, see Patent Document 2) has been published, but there is a drawback that the activity in the neutral range is weak.

JP 2002-171934 A (2nd page) JP 2003-171294 A (pages 2-5)

  An object of the present invention is to provide a thrombus formation inhibitor that can be used for a wide range of foods and drinks and foods and drinks containing the same.

  The present inventors have found that the extract of Gymnema has an excellent thrombus formation inhibitory effect as a result of diversified research and studies for the purpose of searching for a thrombus formation inhibitory component using various natural plants, and completed the present invention. I let you.

From the results of the platelet aggregation test, it was found that the thrombus formation inhibitor containing the extract of Gymnema obtained in the present invention has a high effect of suppressing the formation of platelet aggregates.
Gymnema, in particular, is safe because it has no side effects that cause bleeding in the body, unlike conventional drugs because it is derived from a natural plant that humans have been using in their daily diet.
The present invention uses a thrombus formation inhibitor containing an extract of Gymnema in various foods and beverages, pharmaceuticals, etc., and suppresses thrombus formation to prevent cerebral hemorrhage, cerebral infarction, myocardial infarction, arteriosclerosis and coronary artery disease. Cardiovascular diseases can be prevented.

  The Gymnema used in the present invention is a scientific name: Gymnema Sylvestre, and is native to India, and is a creeping plant of Asperpidaceae that is widely distributed from tropical to subtropical regions such as Indonesia and southwestern China. From the standpoint of plant taxonomy, it is a plant belonging to the same family as Gaigamo, Ikema, Milkweed, etc. growing in Japan.

  In the present invention, one or two or more kinds selected from the group of leaves, stems, and vines are usually used as the gymnema part. The form is not particularly limited. In the case of powder, it can be used as it is, but it is preferable that the water-insoluble component is removed by extraction with water or the like.

The extraction method is not particularly limited, such as extraction solvent, extraction temperature, etc. As the extraction solvent, water, bases, acids, alcohols, other non-organic solvents such as saline can be used, preferably One or more selected from the group of water, base, acid, and alcohol.
When an acid or base is used as the extraction solvent, it is preferable to neutralize the extract. The salt produced by the neutralization reaction can be removed by a known method such as dialysis or gel filtration. When water is used as the extraction solvent, the neutralization reaction as described above is not necessary, and it is not necessary to remove the generated salt. Therefore, it is more preferable to use water.
The acid used at this time is not particularly limited, and most acids can be used, but one kind selected from hydrochloric acid and sulfuric acid or a combination of both is preferred.
Further, the base is not particularly limited, and most of the bases can be used, but preferably one kind selected from sodium hydroxide and potassium hydroxide or a combination of both.
The concentration of the acid or base used for extraction is not particularly limited, and varies depending on the strength of the acid or base, but it is preferable to use a concentration of 0.01 to 0.5 mol.

  The alcohols in the present invention are not particularly limited, but are preferably those that can be used for the preparation of materials for food and drink, more preferably one selected from ethanol, isopropyl alcohol, propylene glycol, and glycerin. Above, most preferably ethanol.

  Further, in the above extraction, it is preferable to repeat the extraction step once or more for the extraction residue, because the extraction rate is improved and the yield is improved. The solvent used for extraction in this case may be the same, or another solvent may be used.

The above extract can be used as it is, but it is preferable because removal of insoluble substances by filtration or centrifugation increases the effect of inhibiting thrombus formation and widens the application range.
It is preferable to recover the supernatant obtained by adding ethanol after removing the insoluble material as it is or after concentrating the extract, since the effect of inhibiting thrombus formation is further enhanced. Although it does not specifically limit as a density | concentration of ethanol, 10-95% is preferable as a final concentration from the point of a yield and an effect, and 60-90% is still more preferable.
The extract can be used as it is, but if necessary, it can also be used as a dry powder by means such as spray drying or freeze drying.

The thrombus formation suppression in the present invention is not particularly limited, but is preferably antiplatelet aggregation.
In the present invention, antiplatelet aggregation refers to suppression of platelet aggregation, and is also simply referred to as antiplatelet (Antiplatelet). The anti-platelet activity is determined when, for example, a platelet aggregometer (Aggregometer) is used, and a substance that causes aggregation (ADP, epinephrine, collagen, arachidonic acid, etc.) is added to the platelet suspension (Platelet rich plasma). This can be confirmed by a method of measuring the platelet aggregation rate.

The thrombus formation inhibitor of the present invention can be applied to foods and drinks, pharmaceuticals, feeds, and the like, and preferably foods and drinks that can be easily consumed by humans.
The food and drink in the present invention is not particularly limited as long as it is a form that can be taken orally, such as a solution, suspension, powder, or solid molded product. More specifically, instant noodles, retort foods, canned foods, microwave foods, instant soups and miso soups, freeze-dried foods, soft drinks, fruit juice drinks, vegetable drinks, soy milk drinks, coffee drinks, tea drinks Beverages such as powdered beverages, concentrated beverages, nutritional beverages, alcoholic beverages, bread, pasta, noodles, cake mixes, flour products such as fried flour, bread crumbs, rice cakes, caramel, chewing gum, chocolate, cookies, biscuits, cakes, Sweets such as pies, snacks, crackers, Japanese sweets, desserts, sauces, processed tomato seasonings, flavor seasonings, cooking mixes, sauces, dressings, soups, curry and stew seasonings, processing Fats and oils such as butter, margarine and mayonnaise, milk beverages, yogurts, lactic acid bacteria beverages, ice creams, chestnuts Agricultural processing such as dairy products such as mussels, frozen foods, processed fishery products such as fish meat ham and sausage, fish paste products, livestock processed products such as livestock ham and sausage, canned agricultural products, jams and marmalades, pickles, boiled beans and cereals Products, nutritional foods, tablets, capsules and the like.

In this invention, when processing into a thrombus formation inhibitor or food-drinks etc., various nutrient components can be strengthened.
Nutritional ingredients that can be enhanced include vitamins such as vitamin A, vitamin B ₁ , vitamin B ₂ , vitamin B ₆ , vitamin B ₁₂ , vitamin C, vitamin D, vitamin E, niacin (nicotinic acid), pantothenic acid, folic acid, Essential amino acids such as lysine, threonine and tryptophan, minerals such as calcium, magnesium, iron, zinc and copper, and for example, α-linolenic acid, EPA, DHA, evening primrose oil, octacosanol, casein phosphopeptide (CPP), Casein calcium peptide (CCP), water-soluble dietary fiber, insoluble dietary fiber, substances that contribute to human health, such as oligosaccharides, and one or more useful substances approved as other foods and food additives Can be used.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, the following Example does not limit the scope of the present invention.

(Example 1) Preparation 1 of a thrombus formation inhibitor
Gymnema dry powder was pulverized to 40 mesh or less, 2 liters of distilled water was added to 80 g of the powder, and extracted at 55 ° C. for 3 hours. Thereafter, the extract and the residue were separated by filtration. Furthermore, 2 liters of distilled water was added to the residue, and extraction was repeated once more under the same conditions. The extracts were combined and then freeze-dried to obtain 28 g of thrombus formation inhibitor A of the present invention.

(Test Example 1) Confirmation of thrombus formation inhibitory effect The antiplatelet activity of the thrombus formation inhibitor of the present invention was determined to be 400 μl of a healthy person's platelet suspension (platelet rich plasma) using a platelet aggregometer (Aggregometer). After adding 80 μl of the sample, 20 μl of ADP (1 mg / ml solution) was added as a substance causing aggregation, and the platelet aggregation rate after 5 minutes was measured.
Separately, water (sample concentration 0 mg / ml) was added as a control, and the platelet aggregation rate was measured by the same method. From the measured platelet aggregation rate, antiplatelet activity (%) relative to the control was calculated according to the following formula.
Antiplatelet activity (%) = (control platelet aggregation rate−platelet aggregation rate at the time of sample addition) / control platelet aggregation rate × 100
Table 1 shows the results of measurement at sample concentrations of 2.5, 5.0, and 7.5 mg / ml.

  From the results in Table 1 above, it was confirmed that the thrombus formation inhibitor of the present invention exhibits high antiplatelet activity. It was also confirmed that the antiplatelet activity increased proportionally by increasing the extract concentration.

(Example 2) Preparation 2 of thrombus formation inhibitor
Gymnema dry powder was pulverized to 40 mesh or less, 2 liters of distilled water was added to 80 g of the powder, and extracted at 55 ° C. for 3 hours. Thereafter, the extract and the residue were separated by filtration. Further, 2 liters of distilled water was added to the residue, and extraction was repeated once again under the same conditions. The extracts were combined and concentrated under reduced pressure to 200 ml. Ethanol was added to this concentrated solution to prepare 1 liter (final ethanol concentration 80%), and then allowed to stand at 4 ° C. for 24 hours to precipitate insoluble components.
The precipitate and the supernatant are separated by centrifugation, the precipitate is dried under reduced pressure, redissolved in 2 liters of water, filtered to remove insoluble components, and the filtrate is lyophilized to obtain the 80% ethanol-insoluble fraction of the present invention. 7 g of a certain thrombus formation inhibitor B was obtained.
Similarly, the supernatant was processed to obtain 21 g of thrombus formation inhibitor C, which is an 80% ethanol-soluble fraction.

(Test Example 2) Confirmation of thrombus formation inhibitory effect About the thrombus formation inhibitors B and C obtained in Example 2, the antiplatelet activity was measured in the same manner as in Test Example 1 at a sample concentration of 5.0 mg / ml. . The results are shown in Table 2.

  From the results in Table 2 above, it was found that the antiplatelet activity was higher in the soluble fraction with 80% ethanol.

(Example 3) Preparation of thrombus formation inhibitor-containing food (tablet confectionery) 50 g of thrombus formation inhibitor B obtained in Example 2, 30 g of lactose, DHA-containing powder oil (Suncoat DY-5; manufactured by Taiyo Chemical Co., Ltd.) ) 12 g, 4 g of sucrose fatty acid ester, and 4 g of yogurt flavor were mixed and tableted so that 1 tablet would be 300 mg to obtain a thrombus formation inhibitor-containing food or drink (tablet cake) of the present invention.

(Example 4) Preparation of thrombus formation inhibitor-containing beverage (vegetable juice mixed drink) 1 g of thrombus formation inhibitor B obtained in Example 2 and 3 g of guar gum degradation product (Sunfiber R; manufactured by Taiyo Chemical Co., Ltd.) It added, mixed and dissolved in 100 ml of commercially available vegetable juice mixed drinks, and the thrombus formation inhibitor containing food / beverage products (vegetable fruit juice mixed drink) of this invention were obtained.

The embodiment of the present invention and the target product are as follows.
(1) A thrombus formation inhibitor characterized by containing Gymnema or an extract of Gymnema.
(2) The thrombus formation inhibitor as described in (1) above, wherein the extract of Gymnema is extracted with any of water, base, acid, and alcohols.
(3) The thrombus formation inhibitor as described in (1) or (2) above, wherein the Gymnema or Gymnema extract is extracted from Gymnema or Gymnema with water.
(4) The thrombus formation inhibitor according to any one of (1) to (3), wherein the agent is fractionated with ethanol from Gymnema or an extract of Gymnema or Gymnema.
(5) The thrombus formation inhibitor according to any one of (1) to (4), wherein the agent is fractionated as a soluble component with ethanol from Gymnema or an extract of Gymnema or Gymnema.
(6) The thrombus formation inhibitor according to (5) above, wherein the ethanol concentration when fractionated with ethanol is 10 to 95% as a final concentration, and is a soluble fraction thereof.
(7) The thrombus formation inhibitor according to (5) or (6) above, wherein the ethanol concentration when fractionated with ethanol is 60 to 90% as a final concentration, and is a soluble fraction thereof.
(8) The thrombus formation inhibitor according to any one of (1) to (7) above, wherein the thrombus formation inhibitory action is an action of inhibiting platelet aggregation.
(9) Food / beverage products containing the thrombus formation inhibitor in any one of said (1)-(8).
(10) A pharmaceutical comprising the thrombus formation inhibitor according to any one of (1) to (8).
(11) A feed comprising the thrombus formation inhibitor according to any one of (1) to (8).

  The thrombus formation inhibitor containing the Gymnema or Gymnema extract obtained in the present invention has a high antiplatelet activity that inhibits the adhesion of platelets to the blood vessel wall and suppresses the formation of platelet aggregates. It can be used in medicines to prevent cardiovascular diseases such as cerebral hemorrhage, cerebral infarction, myocardial infarction, arteriosclerosis and coronary artery disease by suppressing the production of platelet aggregates.

Claims (4)

A thrombus formation inhibitor characterized by containing Gymnema or an extract of Gymnema. The thrombus formation inhibitor according to claim 1, wherein the extract of Gymnema is extracted using at least one selected from the group consisting of water, base, acid, and alcohols. The thrombus formation inhibitor according to claim 1 or 2, wherein the extract of Gymnema is a soluble fraction further fractionated with ethanol. Food / beverage products containing the thrombus formation inhibitor in any one of Claims 1-3.