JP2005220100A - Anti-ageing agent, platelet aggregation inhibitor, antioxidant, anti-allergic agent, skin cosmetic, and food and drink - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain an anti-ageing agent, a platelet aggregation inhibitor, an antioxidant, an anti-allergic agent, a skin cosmetic, and food and drink each containing a natural extract. <P>SOLUTION: The anti-ageing agent, platelet aggregation inhibitor, antioxidant, anti-allergic agent, skin cosmetic, and food and drink contain each an extract from Astragalus complanatus R.Br. as an effective ingredient. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to an anti-aging agent, a platelet aggregation inhibitor, an antioxidant, an antiallergic agent, skin cosmetics, and foods and drinks containing a highly safe plant extract as an active ingredient.

  The epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrices such as elastin, collagen, and hyaluronic acid that are outside these cells and support the skin structure. In young skin, moisture retention, flexibility, elasticity and the like are ensured by maintaining the constancy of the interaction between these skin tissues, and the skin is maintained in a fresh and fresh state.

  However, when there is an influence of certain external factors such as irradiation of ultraviolet rays, significant drying of air, excessive skin washing, etc., or when aging progresses, the production amount of collagen, which is a main component of the extracellular matrix, is increased. It causes a decrease in elasticity due to cross-linking. As a result, the skin retains its moisturizing function and elasticity, and the keratin begins to exfoliate abnormally. Therefore, the skin loses its tension and gloss, and exhibits an aging phenomenon such as rough skin and wrinkles. As described above, changes associated with skin aging, that is, wrinkles, dullness, disappearance of texture, decrease in elasticity, and the like are associated with a decrease / denaturation of dermal matrix components such as collagen.

  Therefore, it is considered that aging of the skin can be prevented or improved by promoting the production of collagen in dermal fibroblasts. As a plant extract having a collagen production promoting action, an extract from pentagon ( Patent Document 1), an extract from a lotus germ (Patent Document 2), an extract from a five-finger peach (Patent Document 3), and the like are known.

  In addition, the cause of skin aging accompanying aging is that the secretion amount of estrogen, a kind of female hormone, decreases. In other words, estrogen is deeply involved in maintaining the health of adult women, and its lack of secretion leads to various medical illnesses, as well as the cause of unfavorable skin changes such as skin sensitivities, decreased elasticity and decreased moisture. It is known to be. Therefore, a substance that exerts the same action as estrogen is transcutaneously or orally administered to women in menopause and after the estrogen secretion declines. For this purpose, steroidal estrogens, nonsteroidal estrogens, flavone compounds and the like have been used as estrogen-like agents.

  On the other hand, platelets agglutinate and activate to cause physiological hemostasis and pathologically thrombus formation, and platelet aggregation is involved in the progression of arteriosclerosis, cancer metastasis, inflammatory diseases, etc. It is believed that For this reason, it is considered that the above-mentioned diseases can be prevented or treated with substances that inhibit and suppress platelet aggregation. As a plant extract having an anti-inflammatory action, an extract from Bergenia (Patent Document 4), wisteria Extracts from tea (Patent Document 5), extracts from red snow tea (Patent Document 6), and the like are known.

In addition, active oxygen is generated by an energy metabolism process in living cells, and superoxide (that is, superoxide anion generated by one-electron reduction of oxygen molecules) (.O ₂ ⁻ ), hydrogen peroxide (H ₂ O _2). ), Hydroxy radical (.OH) and the like. In the living body, radicals generated first based on oxygen are superoxide, and other radicals such as hydroxy radicals are generated via superoxide. Superoxide is converted into hydrogen peroxide by the action of superoxide dismutase (hereinafter referred to as “SOD”) produced in cells.

  The amount of SOD decreases with aging, and the intracellular concentration of superoxide increases due to the decrease in SOD, so that the activity of catalase, which is a detoxifying enzyme for active oxygen, decreases, and superoxide causes damage to the living body. Become. For example, tissue disorders such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke, cataract, diabetes, arteriosclerosis, stiff shoulders, coldness, skin blemishes and wrinkles are brought about. As an SOD-like agent for preventing or treating such a disorder, an extract from urgon (Patent Document 7), a spinach pear fruit juice (Patent Document 8), and the like are known.

  By the way, in recent years, allergic diseases that develop through allergic reactions such as hay fever, allergic rhinitis, bronchial asthma, and atopic dermatitis have become one of the modern diseases seen in a wide age group from children to adults. The allergic reaction that causes it is divided into type I to type IV. For example, in type I allergy (immediate type allergy) represented by allergic rhinitis and bronchial asthma, chemical mediators such as histamine, leukotriene, prostaglandin are released from mast cells and basophils. Causes an inflammatory response with symptoms such as rhinitis and asthma. Further, in type IV allergy (delayed type allergy) typified by allergic contact dermatitis, cytokines released by T cells activate and accumulate eosinophils and macrophages to cause an inflammatory reaction.

  Such allergic reactions and accompanying inflammatory reactions are caused by histamine release in the body, platelet aggregation, generation of active oxygen and radicals, and the like.

Histamine release is a phenomenon in which histamine in mast cells is released extracellularly, and an inflammatory reaction is caused by the released histamine. For this reason, attempts have been made to prevent or treat allergic diseases with substances that inhibit or suppress histamine release. As herbal medicines having an action of suppressing the release of histamine, extracts from plants belonging to the genus Choerospondias (Patent Document 9) and extracts from the root of Lantana (Patent Document 10) have been known so far. It has been.
JP 2002-226323 A JP 2002-29980 A JP 2003-95970 A Japanese Patent Laid-Open No. 11-209299 JP 2001-97873 A JP 2003-212789 A Japanese Unexamined Patent Publication No. 64-50877 Japanese Patent Laid-Open No. 3-83548 JP 2003-55245 A Japanese Patent Laid-Open No. 2002-179583

  The object of the present invention is to first find a natural product having a collagen production promoting action and / or an estrogen-like action, and to provide an anti-aging agent comprising the same as an active ingredient.

  A second object of the present invention is to find a natural product having an inhibitory action on platelet aggregation, and to provide a platelet aggregation inhibitor containing the same as an active ingredient.

  A third object of the present invention is to find an active oxygen scavenging action and / or radical scavenging action from natural products, and to provide an antioxidant containing the same as an active ingredient.

  A fourth object of the present invention is to find a natural product having an inhibitory action on histamine release, and to provide an antiallergic agent comprising the same as an active ingredient.

  A fifth object of the present invention is to find a natural product having a collagen production promoting action or an estrogen-like action from natural products, and to provide a skin cosmetic containing the same.

  Sixth, the present invention finds a natural product having a collagen production promoting action, an estrogen-like action, a platelet aggregation inhibitory action, an active oxygen scavenging action, a radical scavenging action or a histamine release inhibiting action, and a food and drink containing the same The purpose is to provide goods.

  In order to solve the above-mentioned problems, the anti-aging agent, platelet aggregation inhibitor, antioxidant, anti-allergic agent, skin cosmetics and foods and beverages of the present invention are characterized by containing an extract from Root Genge as an active ingredient. To do.

  In the anti-aging agent of the present invention, the extract preferably has a collagen production promoting action and / or an estrogen-like action. Moreover, in the antioxidant of this invention, it is preferable that the said extract has an active oxygen scavenging action and / or a radical scavenging action. Furthermore, in the antiallergic agent of the present invention, the extract preferably has a histamine release inhibitory action.

  The anti-aging agent, platelet aggregation inhibitor, antioxidant and anti-allergic agent of the present invention can be easily produced by an operation such as simple extraction from a natural plant that is readily available.

  In addition, according to the anti-aging agent, platelet aggregation inhibitor, antioxidant or antiallergic agent of the present invention, wrinkle formation and reduced elasticity associated with skin aging; progression of arteriosclerosis associated with platelet aggregation, cancer Metastases or inflammatory diseases, etc .; tissue damage such as rheumatoid arthritis and Behcet's disease associated with increased intracellular active oxygen concentration, myocardial infarction, stroke, cataract, diabetes, arteriosclerosis, stiff shoulders, coldness, skin spots, wrinkles, etc. Disorders, inflammatory diseases, allergic diseases and the like; or inflammatory diseases and allergic diseases associated with histamine release can be prevented and improved.

  Furthermore, the skin cosmetics and foods and beverages of the present invention can be easily produced by an operation such as simple extraction from a natural plant that is readily available. And according to the skin cosmetics of the present invention, it is possible to prevent and improve skin aging, etc., and according to the food and drink of the present invention, skin aging, inflammatory diseases associated with platelet aggregation, etc. Rheumatoid arthritis associated with an increase in active oxygen concentration or allergic diseases associated with histamine release can be prevented / ameliorated.

Hereinafter, the present invention will be described in detail.
[Anti-aging agent, platelet aggregation inhibitor, antioxidant, antiallergic agent]
The anti-aging agent, the platelet aggregation inhibitor, the antioxidant and the antiallergic agent of the present invention contain an extract from Root Genge as an active ingredient.

  Here, in the present invention, the “extract from tsurugenge” includes an extract obtained from tsurugenge as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or Any of these crudely purified products or purified products is included.

  The plant used as an extraction raw material is Astragalus complanatus R.Br. Vulgaris is a legume plant and a perennial. It grows naturally in mountains such as China and Inner Mongolia, and can be easily obtained from these areas. The seeds are said to be especially called Xiaenshi, and are used as a component of traditional Chinese medicine to improve liver and kidney function decline and dizziness.

  There are no particular limitations on the part of the vine that can be used as the raw material for extraction, for example, flowers, flower spikes, peels, fruits, cones, stems, leaves, branches, branches and leaves, trunks, bark, rhizomes. , Root bark, roots, seeds, whole plants, etc., and one or more of these can be used as the extraction raw material, but it is particularly preferable to use seeds.

  As for the pickaxe used as an extraction raw material, it is suitable to dry and pulverize immediately after collection. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pre-processing, such as degreasing, with nonpolar solvents, such as hexane and benzene, you may use as an extraction raw material. By performing pretreatment such as degreasing, extraction with a polar solvent of vine leaves can be performed efficiently.

  Details of anti-aging substances, platelet aggregation inhibitory substances, antioxidant substances or antiallergic substances contained in extracts from vines are unknown, but anti-aging forms from vines by means of extraction methods commonly used for plant extraction. An extract having an action, a platelet aggregation inhibitory action, an antioxidant action or an antiallergic action can be obtained.

  As an extraction solvent, it is preferable to use a polar solvent, for example, water, a hydrophilic organic solvent, etc. are mentioned, These can be used individually or in combination of 2 or more types.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, superheating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, and the like. Therefore, the water that can be used as the extraction solvent in the present invention includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using the liquid mixture of water and a lower aliphatic alcohol, the mixing ratio of water and a lower aliphatic alcohol can be set to 7: 3 to 2: 8 (mass ratio).

  The extraction treatment is not particularly limited as long as the soluble component contained in the vine genus can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extract is obtained by immersing the vine root in 1 to 20 times the extraction solvent (mass ratio) of the extraction raw material, extracting the soluble components at room temperature or under reflux, and then filtering to remove the extraction residue. Can be obtained. The obtained extract is diluted, concentrated, dried, purified, etc. according to a conventional method in order to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crude purified product or a purified product thereof. Processing may be performed.

  Purification can be performed by, for example, activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, and the like. The obtained extract can be used as it is as an active ingredient of an anti-aging agent, anti-inflammatory agent, antioxidant or anti-allergic agent, but a concentrated solution or a dried product is easier to use.

  Tsurugenge extract has anti-aging action, platelet aggregation inhibitory action, antioxidant action or anti-allergic action, and it is formulated in a conventional manner as it is or with other molding aids in the form of powder, granules, tablets And can be provided as any dosage form, and can be used as a skin preparation, an internal solution, a solid preparation, an injection, a suppository, and the like.

  When formulating the obtained vine extract, a pharmaceutically acceptable carrier such as dextrin or cyclodextrin and other optional auxiliaries can be added for easy storage and handling. Examples of the auxiliary agent that can be used together with the extract from the pickaxe include excipients, binders, disintegrants, lubricants, stabilizers, flavoring and flavoring agents, and the like.

  The extract from the pickaxe obtained as described above has an anti-aging effect, a platelet aggregation inhibitory effect, an antioxidant effect or an anti-allergic effect. It can be used as an agent, antioxidant or antiallergic agent.

  Here, the anti-aging effect of the extract from the vine is exerted based on, for example, a collagen production promoting action and / or an estrogen-like action. However, the anti-aging action which the extract from a vine leaves has is not necessarily limited to the anti-aging action exhibited based on the said action.

  Moreover, the antioxidant action which the extract from a rugogenge has is exhibited based on, for example, an active oxygen scavenging action and / or a radical scavenging action. However, the antioxidant action which the extract from a vine leaves has is not limited to the antioxidant action exhibited based on the said action. Here, “active oxygen” includes superoxide, hydrogen peroxide, hydroxy radical, singlet oxygen and the like. The term “radical” means a molecule or atom having one or more unpaired electrons, and includes superoxide, hydroxy radical, DPPH and the like.

  Furthermore, the antiallergic action of the extract from vine leaves is exhibited based on, for example, histamine release inhibiting action. However, the antiallergic action of the extract from the vine is not limited to the antiallergic action exhibited based on the above action.

  The anti-aging agent, platelet aggregation inhibitor, antioxidant or anti-allergic agent of the present invention is optionally extracted from other natural extracts having anti-aging action, platelet aggregation inhibiting action, antioxidant action or anti-allergic action. It can be used as an active ingredient by blending products.

  The anti-aging agent of the present invention can prevent skin aging through a collagen production promoting action and / or an estrogen-like action of a vine extract, which is an active ingredient. Therefore, the anti-aging agent of the present invention effectively prevents and improves skin wrinkles, dullness, disappearance of texture, loss of elasticity, etc. due to decrease in collagen and / or decrease in estrogen secretion in dermal fibroblasts. can do. However, the anti-aging agent of the present invention can be used for all uses other than these uses, which are meaningful in suppressing collagen production and / or exhibiting the same action as estrogen.

  In addition, the platelet aggregation inhibitor of the present invention is capable of treating various diseases associated with platelet aggregation, such as progression of arteriosclerosis, cancer metastasis, inflammatory diseases, etc. It can be effectively prevented and treated. However, the platelet aggregation inhibitor of the present invention can be used for all applications other than these applications that are meaningful for inhibiting platelet aggregation.

  Furthermore, the antioxidant of the present invention increases the active oxygen concentration in the body (for example, an increase in the active oxygen concentration due to excessive production of active oxygen in the body, the elderly, etc.) through the antioxidant action of the vine extract, which is an active ingredient. Diseases caused by SOD action increase, etc., such as tissue disorders such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke, cataract, diabetes, arteriosclerosis, stiff shoulders, coldness, skin spots / wrinkles, etc. Can be effectively prevented and improved. However, the antioxidant of the present invention can be used for all uses other than these uses, which are meaningful for eliminating active oxygen.

  The antiallergic agent of the present invention can effectively prevent and improve allergic diseases associated with the release of histamine through the histamine release inhibitory action of the vine extract, which is an active ingredient. However, the antiallergic agent of the present invention can be used for all uses other than these uses, which are meaningful for suppressing the release of histamine.

[Skin cosmetic]
The skin cosmetic of the present invention contains an extract from Astragalus complanatus R. Br. As an active ingredient.

  An extract from vine leaves has an anti-aging effect and is excellent in usability and safety when applied to the skin, and is therefore suitable for blending into skin cosmetics. In this case, the extract from the vine tree may be blended as it is, or an anti-aging agent formulated from the extract from the vine tree may be blended.

  There are no particular limitations on the skin cosmetics that can be blended with the extract from the vine, but specific examples include ointments, creams, emulsions, lotions, packs, jellies, lip balms, lipsticks, bath preparations, etc. is there.

  The compounding agent of the extract from the vine in the skin cosmetics of the present invention can be appropriately adjusted according to the type of skin cosmetics, physiological activity of the extract, etc., but the suitable compounding ratio is converted to a standard extract. And about 0.005 to 10% by mass.

  The skin cosmetics of the present invention are various main agents and auxiliaries usually used in the production of the skin cosmetics, for example, astringents, bactericides / antibacterial agents, as long as they do not interfere with the anti-aging action of the extract from the pickaxe , Whitening agents, ultraviolet absorbers, moisturizers, cell activators, anti-inflammatory / anti-allergic agents, antioxidant / active oxygen scavengers, and the like can be used in combination. When used in this way, it becomes a more general product, and thereby a synergistic effect with other active ingredients used in combination may lead to a superior effect than would normally be expected.

[Food and Drink]
The food / beverage products of this invention contain the extract from a straugeus (Astragalus complanatus R.Br.) as an active ingredient.

  Extracts from vines have anti-aging effects, platelet aggregation-inhibiting effects, antioxidant effects or anti-allergic effects, and are excellent in safety, so they are general foods, health foods, health functional foods or nutritional supplements. It is suitable for blending into any food or drink such as food. In this case, an extract from the vine genus may be blended as it is, or an anti-aging agent, a platelet aggregation inhibitor, an antioxidant or an anti-allergic agent formulated from an extract from the vine genus may be blended.

  When an extract from the above vinegar, an anti-aging agent, a platelet aggregation inhibitor, an antioxidant or an antiallergic agent is added to foods and drinks, the amount of the active ingredient in them is determined in consideration of the purpose of use, gender, symptoms, etc. However, it is preferable that the extract intake per day for adults is about 1 to 1000 mg in consideration of the general intake of the food and drink to be added.

  Foods and drinks that can be blended with the above extract from Root Genge, Anti-aging Agent, Platelet Aggregation Inhibitor, Antioxidant or Antiallergic Agent are various actions that the extract from Root Genge has, for example, anti-aging effect, platelet aggregation inhibition It is not particularly limited as long as it does not hinder the action, antioxidant action, antiallergic action and the like.

  Specific examples include beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks (including concentrated concentrates and powders for adjustment of these drinks); frozen desserts such as ice cream, ice sherbet and shaved ice; Noodles such as udon, harusame, gyoza skin, sweet husk, Chinese noodles, instant noodles, etc .; sweets such as candy, chewing gum, candy, gum, chocolate, tablet confectionery, snack confectionery, biscuits, jelly, jam, cream, baked confectionery Fishery and livestock processed foods such as kamaboko, ham and sausage; dairy products such as processed milk and fermented milk; processed oils and fats such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream and dressing; sauces, sauce Seasonings such as soups, stews, salads, prepared dishes, pickles; various other forms of health and nutrition Supplementary foods: Can be manufactured by blending the above anti-aging agent, anti-inflammatory agent, antioxidant or antiallergic agent into tablets, capsules, drinks, etc. Things can be used together.

  The anti-aging agent, platelet aggregation inhibitor, antioxidant and anti-allergic agent of the present invention are preferably applied to humans, but as long as the desired action and effect are exhibited, the anti-aging agent, platelet aggregation inhibitor, antioxidant and anti-allergic agent It can also be applied to animals. For example, the anti-aging agent, platelet aggregation inhibitor, antioxidant, or antiallergic agent of the present invention is added to pet foods such as dog food and cat food, and livestock feed to prevent and treat various diseases of pet animals and livestock. You can also.

Hereinafter, although a manufacture example, a test example, and a compounding example are shown and this invention is demonstrated concretely, this invention is not limited to each following example at all.
[Production Example 1] Manufacture of Tsurugenge extract To 1000 g of Tsurugenge seeds, 3000 mL of an extraction solvent was added, followed by heating and extraction under reflux for 1 hour, and the extraction operation was repeated twice. The obtained extract was concentrated under reduced pressure, and further dried to obtain a vine extract. Table 1 shows the yield of the extract when the above extraction is performed with various concentrations of the extraction solvent.

[Table 1]
Extraction solvent extract yield (% by mass)
Water 11.2
30% ethanol 10.3
50% ethanol 9.6
70% ethanol 5.5

[Test Example 1] Collagen production inhibitory action test The genus extract (samples 1 to 4) obtained in Production Example 1 is as follows according to the method of Webster et al. (Anal. Biochem., Vol. 96, 220, 1979). The collagen production inhibitory action was tested as described above.

Human fibroblasts were seeded in a 24-well plate and cultured in a sample-added medium (sample concentration: 100 ppm; μg / mL) at 37 ° C. and 5% CO ₂ -95% air for several days. β-aminopropionitrile and [ ³ H] -proline were added and further cultured for 24 hours.

  A pepsin / acetic acid solution is added to the whole culture solution and digested for 16 hours under conditions of 4 ° C., then a carrier is added to the digested solution and precipitated with a 0.7 mol / L saline solution, and further under neutral conditions. Redissolved and reprecipitated with a 4.2 mol / L saline solution. The obtained precipitate was washed with 20% ethanol, and then the radioactivity of the precipitate was measured.

Collagen production promotion rate was calculated with the radioactivity when no sample was added as 100%.
Table 2 shows the collagen production promotion rate (%) of each sample.
[Table 2]
Sample extract collagen production promotion rate (%)
1 Water extract 91.1
2 30% ethanol extract 227.0
3 50% ethanol extract 141.0
4 70% ethanol extract 66.7
From the results shown in Table 2, an excellent collagen production inhibitory action was observed particularly in the vine extract extracted with 30% ethanol or 50% ethanol.

[Test Example 2] Estrogen-like action test The method of Thomas et al. (In Vitro Cell. Dev. Biol.) Was used to examine the effect of eavesman-dependent cell proliferation on the vine extract (samples 1 to 4) obtained in Production Example 1. 28A, 595 to 602, 1992), estrogen-like action was tested as follows.

MCF-7 cells derived from human breast cancer are cultured in a 75 cm ² flask until confluent, and the MCF-7 cells are collected by trypsin treatment. 10% FBS (activated charcoal), 1% NEAA and 1 mM sodium pyruvate And 3 × 10 ⁴ cells / mL using a MEM medium containing no phenol red (hereinafter referred to as “MEM medium”).

The prepared MCF-7 cells were seeded at 0.9 mL each in a 24-well plate and cultured under conditions of 37 ° C. and 5% CO ₂ -95% air in order to establish them. Six hours later (day 0), 100 μL of a sample solution prepared in MEM medium to a concentration 10 times the final concentration (125 ppm; μg / mL) was added to the plate, and the culture was continued.

  On the sixth day from the start of the culture, the medium was replaced with a MEM medium containing 0.97 mmol / L of MTT, and cultured for 2 hours. Then, the medium was replaced with isopropanol, and blue formazan produced in the cells was extracted. About isopropanol containing the extracted blue formazan, the light absorbency of 570 nm with the absorption maximum of blue formazan was measured.

  In order to correct the influence of adherent cells, the absorbance at 650 nm was also measured at the same time, and the difference between the two absorbances was taken as a value proportional to the amount of blue formazan produced (the absorbance in the formula below is this corrected absorbance). . As a positive control, 0.02 ppm ethinylestradiol was used. The strength of the estrogen-like action (estrogen-dependent growth action) was calculated by the following equation, assuming that the absorbance when no sample was added was 100%.

Estrogen-like action rate (%) = A / B × 100
In the above formula, “A” represents “absorbance when the sample is added” and “B” represents “absorbance when the sample is not added”.
The results of this test are shown in Table 3.

[Table 3]
Sample extract estrogen-like action rate (%)
1 Water extract 114.7
2 30% ethanol extract 125.5
3 50% ethanol extract 117.9
4 70% ethanol extract 121.6
From the results in Table 3, excellent estrogen-like action was observed in the vine extract extracted using various extraction solvents.

Test Example 3 Platelet Aggregation Inhibitory Action Test The thruge extract (Samples 1 to 4) obtained in Production Example 1 was tested for platelet aggregation inhibitory action as follows.

  1/10 volume of 77 mM EDTA was added to the blood of Japanese white rabbit, and the precipitate was removed by centrifugation at 1000 rpm for 10 minutes. The supernatant was centrifuged at 2100 rpm for 10 minutes, and the precipitated platelets were collected. The obtained platelets were suspended in a platelet washing solution and centrifuged at 2100 rpm for 10 minutes. The precipitated platelets were collected and suspended in a platelet suspension so that the platelet count was 300,000 / μL.

  1 μL of calcium chloride solution was added to 223 μL of the washed platelet suspension prepared in this manner, and incubated at 37 ° C. for 1 minute. 1 μL of the sample solution was added thereto, and the mixture was further incubated for 2 minutes, and then stirred for 1 minute. Next, 25 μL of a 10 ppm (μg / mL) collagen solution is added as an aggregation inducer, incubated for 10 minutes at 37 ° C., and then the aggregation rate A is measured using a platelet aggregation measuring apparatus (PAM12CL, manufactured by Mevanics Co., Ltd.). did.

  Only the solvent of the sample solution was added instead of the sample solution, and the same operation as described above was performed to measure the aggregation rate B, and the platelet aggregation inhibition rate was determined by the following formula.

Platelet aggregation inhibition rate (%) = {(B−A) / B} × 100
In the above formula, “A” indicates “aggregation rate at the time of adding an aggregation initiator and adding a sample solution”, and “B” indicates “aggregation rate at the time of adding an aggregation initiator and not adding a sample solution”.

The platelet aggregation rate was measured by gradually reducing the concentration of the sample solution, and the sample concentration IC ₅₀ (ppm; μg / mL) at which the platelet aggregation inhibition rate was 50% was determined by interpolation.
The results of this test are shown in Table 4.

[Table 4]
Sample extract IC ₅₀ (ppm)
1 Water extract 1923
2 30% ethanol extract 5100
3 50% ethanol extract-
4 70% ethanol extract −
From the results in Table 4, an excellent inhibitory effect on platelet aggregation was observed in the vine extract extracted with water or 30% ethanol.

[Test Example 4] Superoxide scavenging action test (NBT method)
For the vine extract (samples 1 to 4) obtained in Production Example 1, the superoxide scavenging action was tested as follows.

Take 0.1 mL each of 3 mM xanthine, 0.05 M Na ₂ CO ₃ buffer (pH 10.2), 3 mM EDTA, BSA solution and 0.75 mM NBT in a test tube. Added and left at 25 ° C. for 10 minutes.

  Next, a xanthine oxidase solution was added, stirred rapidly, and allowed to stand at 25 ° C. for 20 minutes. Thereafter, 0.1 mL of 6 mM copper chloride was added to stop the reaction, and the absorbance at a wavelength of 560 nm was measured. Even when the enzyme solution was not added, the same operation and absorbance were measured, and the same measurement was performed when distilled water was added without adding the sample solution. And the superoxide erasure | elimination rate was calculated | required by following Formula.

Erasure rate (%) = {1− (A−B) / (C−D)} × 100
In the above formula, “A” is “absorbance when enzyme solution is added and sample solution is added”, “B” is “absorbance when enzyme solution is not added and sample solution is added”, and “C” is “enzyme solution is added” , “Absorbance when no sample solution is added”, and “D” indicates “absorbance when no enzyme solution is added and no sample solution is added”.

The above-mentioned erasure rate was measured by gradually reducing the sample concentration, and the sample concentration IC ₅₀ (ppm; μg / mL) at which the superoxide erasure rate was 50% was determined by interpolation.
The results of this test are shown in Table 5.

[Table 5]
Sample extract IC ₅₀ (ppm)
1 Water extract 69.4
2 30% ethanol extract 54.6
3 50% ethanol extract 49.1
4 70% ethanol extract 41.3
From the results shown in Table 5, an excellent superoxide scavenging action was observed in the vine extract extracted using various extraction solvents.

[Test Example 5] Radical scavenging action test The scavenge extract (samples 1 to 4) obtained in Production Example 1 was tested for radical scavenging action as follows.

3 mL of the sample solution was added to 3 mL of 1.5 × 10 ⁻⁴ M DPPH ethanol solution, and the container was immediately sealed, shaken and allowed to stand for 30 minutes. Thereafter, the absorbance at a wavelength of 520 nm was measured. As a control, the same operation was performed using a solvent in which the sample solution was dissolved instead of the sample solution, and the absorbance at a wavelength of 520 nm was measured. Further, as a blank, after adding 3 mL of the sample solution to ethanol, the absorbance at a wavelength of 520 nm was measured immediately. From each measured absorbance, the radical scavenging action rate was calculated by the following formula.

Erasure rate (%) = {1− (B−C) / A} × 100
In the above formula, “A” represents “absorbance of control”, “B” represents “absorbance when a sample solution is added”, and “C” represents “absorbance of blank”.

The above-mentioned erasure rate was measured by decreasing the sample concentration stepwise, and the sample concentration IC ₅₀ (ppm; μg / mL) at which the DPPH radical erasure rate was 50% was determined by interpolation.
The results of this test are shown in Table 6.

[Table 6]
Sample extract IC ₅₀ (ppm)
1 Water extract> 100
2 30% ethanol extract 104.4
3 50% ethanol extract 75.1
4 70% ethanol extract 46.2
From the results shown in Table 6, an excellent radical scavenging action was observed in the vine extract extracted using various extraction solvents, particularly the vine extract extracted with 50% ethanol or 70% ethanol.

[Test Example 6] Hydrogen peroxide scavenging action test The hydrogen peroxide scavenging action test was conducted on the vine extract (samples 1 to 4) obtained in Production Example 1 as follows.

  10 μL of a sample solution was added to 10 μL of a hydrogen peroxide standard solution (concentration: 1.5 mM), and incubated at 37 ° C. for 20 minutes. Then, a coloring reagent (DA-64 (manufactured by Wako Pure Chemical Industries, Ltd.) was added to 10 mM, Triton X- 1 mL of a peroxidase solution (100 units / mL: manufactured by Wako Pure Chemical Industries, Ltd.) was added to 0.1 M PIPES buffer (pH 7.0) containing 0.5% of 100 (trade name, manufactured by Rohm & Hass Co.), 2.98 mL of the total amount adjusted to 100 mL) was added, incubated at 37 ° C. for 5 minutes, and then the absorbance at a wavelength of 727 nm was measured. The same operation and absorbance measurement were performed when no hydrogen peroxide standard solution was added, and the same measurement was also performed when distilled water was added without adding the sample solution. And the elimination rate of hydrogen peroxide was calculated | required by following Formula.

Erase rate (%) = {1− (A−B) / (C−D)} × 100
In the above formula, “A” is “absorbance when hydrogen peroxide standard solution is added and sample solution is added”, “B” is “absorbance when hydrogen peroxide standard solution is not added and sample solution is added”, “C "Represents" absorbance when hydrogen peroxide standard solution is added and no sample solution ", and" D "represents" absorbance when hydrogen peroxide standard solution is not added and sample solution is not added ".

The above-mentioned erasure rate was measured by gradually decreasing the sample concentration, and the sample concentration IC ₅₀ (ppm; μg / mL) at which the erasure rate of hydrogen peroxide was 50% was determined by interpolation.
The results of this test are shown in Table 7.

[Table 7]
Sample extract IC ₅₀ (ppm)
1 Water extract 34.0
2 30% ethanol extract 14.7
3 50% ethanol extract 11.4
4 70% ethanol extract 10.1
From the results shown in Table 7, an excellent hydrogen peroxide scavenging action was observed in the vine extract extracted using various extraction solvents.

[Test Example 7] Histamine release inhibitory effect test The histamine extract (samples 1 to 4) obtained in Production Example 1 was tested for histamine release inhibitory action as follows. In addition, when intracellular histamine is released, hexosaminidase is also released at the same time, so histamine release inhibitory action was evaluated using hexosaminidase release as an index.

A medium (15% FBS-added S-MEM medium) placed in a 25 mL culture flask was seeded with 1.0 × 10 ⁶ RBL-2H3 cells and cultured under conditions of 37 ° C., 5% CO ₂ -95% air. Cultured for days.

The cells were then collected by trypsinization and centrifugation (800 rpm, 4 minutes). The obtained cells were suspended in the medium at 4.0 × 10 ⁵ cells / mL, and mouse monoclonal anti-dinitrophenyl group IgE (DNP-Specific IgE) was added thereto at a concentration of 0.5 μg / mL. This cell suspension was seeded at 100 μL per well of a 96-well plate and cultured at 37 ° C. and 5% CO ₂ -95% air for 24 hours.

  After completion of the culture, the medium in each well was removed and washed twice with Silaganian buffer. Next, 30 μL of Siraganian buffer and 10 μL of the sample solution were added and incubated at 37 ° C. for 10 minutes. Then, 10 μL of dinitrophenylated bovine serum albumin (DNP-BSA) was added, and further incubated at 37 ° C. for 15 minutes. Thereafter, 10 μL of the supernatant was transferred to a new 96-well plate under ice-cooling, 10 μL of 1 mmol / L p-nitrophenyl-N-acetyl-β-D-glucosamide solution was added thereto, and incubated at 37 ° C. for 1 hour. did.

After the reaction was completed, 250 μL of a 0.1 mol / L NaCO—Na ₂ HCO ₃ solution was added, and absorbance A at 415 nm was measured with a microplate reader at 650 nm as a control. Absorbance measurement was also performed on a cell supernatant to which a Siraganian buffer was added instead of the sample solution and a 0.1 mol / L NaCO-Na ₂ HCO ₃ solution reacted in the same manner (measured at this time). The absorbance was C). The same operation was carried out for a sample in which a Siraganian buffer was added instead of DNP-BSA (the absorbance measured at this time was D). And the hexosaminidase release inhibitory action rate was calculated | required by following Formula.

Release inhibition rate (%) = [1-{(A-C-D) / (B-D)}] × 100
The sample concentration was decreased stepwise to measure the inhibition rate, and the sample concentration IC ₅₀ (ppm; μg / mL) that inhibits the release of hexosaminidase by 50% was determined by interpolation.
The results of this test are shown in Table 8.

[Table 8]
Sample extract IC ₅₀ (ppm)
1 Water extract-
2 30% ethanol extract> 400
3 50% ethanol extract 322
4 70% ethanol extract 292
From the results shown in Table 8, an excellent histamine release inhibitory action was observed in the vine extract extracted with a water-ethanol extraction solvent, particularly the vine extract extracted with 50% ethanol or 70% ethanol.

[Formulation Example 1]
The following raw materials were uniformly mixed, granulated by a conventional method, and then tableted to produce a tablet-like health food.
Turgenge 50% ethanol extract (Production Example 1) 32 g
50g sugar
Crystalline cellulose 10g
Sucrose fatty acid ester 8g

[Formulation Example 2]
Cyclodextrin Minamata (Seldex SL-20: manufactured by Nippon Food Processing Co., Ltd.) 2.994 g was mixed with 0.006 g of the 50% ethanol extract obtained from Production Example 1, and the cyclodextrin of the tsurugen extract. Inclusions were produced. Using this inclusion and the following raw materials, a soft drink was produced according to a conventional method.
3 g of Cyclodextrin inclusion product
Liquid sugar 10g
87g of water

[Composition Example 3]
The following raw materials were mixed, and a soft capsule-shaped health food was produced according to a conventional method.
C. rugenge 30% ethanol extract (Production Example 1) 90g
Turmeric oleoresin turmeric 25g
113g vegetable oil
Glycerin fatty acid ester 12g
60g beeswax

[Composition Example 4]
A lotion having the following composition was produced by a conventional method.
Tsurugenge 30% ethanol extract (Production Example 1) 2g
Glycerin 3g
1,3-butylene glycol 3g
Oleic acid polyoxyethylene sorbitan (20E.0.) 0.5g
Methyl paraoxybenzoate 0.15g
Citric acid 0.1g
Sodium citrate 0.1g
Oil soluble licorice extract 0.1g
Seaweed extract 0.1g
Xylobiose Mixture 0.5g
Kujin extract 0.1g
Fragrance 0.05g
Purified water remainder (total amount is 100 g)

[Formulation Example 5]
An emulsion having the following composition was produced by a conventional method.
Tsurugenge 30% ethanol extract (Production Example 1) 1g
Jojoba oil 4g
2g olive oil
Squalane 2g
Cetanol 2g
2g glyceryl monostearate
Polyoxyethylene cetyl ether (20E.0.) 2.5g
Oleic acid polyoxyethylene sorbitan (20E.0.) 2g
Twilight extract 0.1g
Ginkgo biloba extract 0.1g
Conchiolin 0.1g
Oat extract 0.1g
Chamomile extract 0.1g
1,3-butylene glycol 3g
Methyl paraoxybenzoate 0.15g
Fragrance 0.05g
Purified water remainder (total amount is 100 g)

The skin aging agent of the present invention is used for the prevention / improvement of skin wrinkle formation associated with a decrease in collagen or estrogen secretion, and the anti-inflammatory agent of the present invention is used for the prevention / improvement of inflammatory diseases associated with platelet aggregation. For the improvement, the antioxidant of the present invention is used for the prevention / improvement of rheumatoid arthritis associated with the increase of intracellular active oxygen concentration, and the anti-allergic agent of the present invention is used for the prevention / reduction of allergic diseases associated with the release of histamine. Can greatly contribute to improvement.

Claims (9)

  An anti-aging agent comprising an extract from Astragalus complanatus R.Br. as an active ingredient.   The anti-aging agent according to claim 1, wherein the extract has a collagen production promoting action and / or an estrogen-like action.   A platelet aggregation inhibitor comprising an extract from Astragalus complanatus R.Br. as an active ingredient.   An antioxidant comprising an extract from Astragalus complanatus R. Br. As an active ingredient.   The antioxidant according to claim 4, wherein the extract has an antioxidant action and / or a radical scavenging action.   An antiallergic agent characterized by containing an extract from Astragalus complanatus R.Br. as an active ingredient.   The antiallergic agent according to claim 6, wherein the extract has a histamine release inhibitory action.   A skin cosmetic containing an extract from Astragalus complanatus R.Br. Food and drink containing an extract from Astragalus complanatus R.Br.