JPH06192114A - Medicine comprising extract of hydrangeae dulcis folium as active ingredient and food and cosmetic containing the same blended therein - Google Patents

Abstract

PURPOSE:To obtain a medicine having antiinflammatory, antiallergic and inhibitory actions on decomposition of hyaluronic acid, and a food and a cosmetic comprising the medicine. CONSTITUTION:The medicine, food and cosmetic comprise an extract of Hydrangeae Dulcis Folium as an active ingredient. The inflammation or swelling of the throat caused by a cold, pollenosis, cough, etc., can be prevented and improved by daily ingestion of this food. The pruritus of the skin, etc., can similarly be improved by using this cosmetic. The extract of the Hydrangeae Dulcis Folium as a strong inhibitor of decomposition of the hyaluronic acid is capable of suppressing the reduction in content of the hyaluronic acid contained in the skin, arterial wall, articular cavity, etc., thereby enhancing the humectant properties and softness of the skin in spite of its indirect action, preventing arteriosclerosis due to aging and contributing to an improvement, etc., in arthritis.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a medicine containing an extract of beetroot tea as an active ingredient, more specifically, a medicine having anti-inflammatory, anti-allergic and hyaluronic acid decomposition-inhibiting effects, and foods and cosmetics containing the same. Regarding

[0002]

2. Description of the Related Art With the development of science and technology, many compounds have been synthesized and drugs having various actions have been found.
For example, a large number of drugs, mainly chemically synthesized products, have been developed as anti-inflammatory agents. And, about 100 years ago, the point of action of aspirin, which has been used clinically, lies in the inhibition of cyclooxygenase, which is the initial enzyme in the process from arachidonic acid to the synthesis of prostaglandins (hereinafter abbreviated as "PG"). Following the clarification, indomethacin and the like, which have a strong medicinal effect by a similar mechanism of action, have been developed.

As an antiallergic agent, disodium cromoglycate (hereinafter referred to as "D
Chemically synthesized products such as SCG) are used as an inhibitor of 5-lipoxygenase that produces leukotriene from arachidonic acid, such as a mugwort extract containing a large amount of caffeic acid.

Although all of the above are related to the components of pharmaceuticals, on the other hand, food materials that are thought to contribute to anti-inflammatory or anti-allergy are also required, but Rakan fruit extract and the like are used only based on the tradition. Is the current situation. In addition, various cosmetic raw materials having anti-inflammatory or antiallergic activity have been searched, but safe and promising ones have not been obtained yet.

On the other hand, as a moisturizer, hyaluronic acid derived from microorganisms and the like is mainly mixed in cosmetics. Recently, allergic effects of tea extract are obtained by using the inhibitory activity against hyaluronidase, which is a hyaluronic acid-degrading enzyme, as an index. Attempts have been made to evaluate activity (Yumie Maeda et al .: Shokuei, Vol. 31, 233-237, 1990),
A correlation between anti-hyaluronidase inhibitory activity and allergic activity has been suggested.

[0006]

[Problems to be Solved by the Invention] For the 21st century, there is a growing demand for prevention rather than treatment of diseases. For example, inflammation and allergies are considered to be an overreaction of the resistance of the human body, but some kind of preventive measures are needed when suffering from the onset of illness. The recent surge in allergic diseases such as hay fever is just amazing. Originally, IgE, which is provided in the human body for the purpose of eliminating parasites, has recently been working on allergens with a decrease in the human body's possession of parasites. We cannot overlook the fact that the intake of many things is increasing. This is because linoleic acid is easily desaturated in the living body to arachidonic acid, and leukotriene produced therefrom causes allergies. As society gets richer, allergies tend to increase due to the reduction of human body possession of parasites and the increase of fat intake, but no sufficiently effective anti-allergic agent has been found. Development is an important issue.

On the other hand, the necessity of maintaining and increasing the hyaluronic acid content in the living body is not limited to the skin problem. The water retention structure by hyaluronic acid also plays an important role in the aorta and joint fluid. As aging is accompanied by a decrease in the hyaluronic acid content of the human body,
It is expected that the need for maintaining the water content retained by hyaluronic acid and thus the flexibility of the skin and blood vessels for the aging society will increase more and more. At present, only hyaluronic acid, which is externally applied as a moisturizer for cosmetics, is of interest, and there are almost no attempts to maintain the hyaluronic acid content in the human body, and thus the water content, which remains an important issue.

The above-mentioned problem can be solved by administering a drug, but it is more preferable that the above-mentioned problem is solved by a food or a cosmetic that is ingested or used on a daily basis. However, until now, promising food materials and cosmetic raw materials having anti-inflammatory, anti-allergic and anti-hyaluronidase activities have not been obtained, and provision of such raw materials and raw materials remains an issue.

[0009]

[Means for Solving the Problems] In order to solve the above problems, the present inventor has selected both enzymes, cyclooxygenase, which is the initial enzyme in the process leading to the synthesis of PG from arachidonic acid, and hyaluronidase, which is a hydrolase of hyaluronic acid. An intensive search was conducted to find a natural product that inhibits and also has antihistamine activity. Here, the purpose of inhibiting hyaluronidase is that inhibition of this enzyme strongly correlates not only with maintaining the level of hyaluronic acid in the human body, but also with anti-inflammatory and anti-allergic activities. This enzyme is activated during inflammation, destroys the connective tissue matrix,
Since inhibition of this enzyme is thought to play a role in enhancing the permeability of cells and blood vessels of the inflammatory system, it is considered that inhibition of this enzyme leads to anti-inflammatory. In addition, hyaluronidase is likely to be involved in a series of processes in mast cells, from the binding of IgE-antigen complex to the receptor to the release (degranulation) of histamine granules. This is because a large morphological change such as degranulation cannot occur without temporary destruction of the water retention structure by hyaluronic acid. In fact, many antihistamines also have hyaluronidase inhibitory activity.

As a result of searching for natural products having the above-mentioned properties, a substance having the above-mentioned three activities in the extract of aquatic tea water was sufficiently found, and the present invention was completed.

That is, the first object of the present invention is to provide a medicine containing an extract of the tea beetle as an active ingredient and selected from an anti-inflammatory agent, an anti-allergic agent and a decomposition inhibitor of hyaluronic acid. Another object of the present invention is to provide foods and cosmetics containing the drug.

The beetroot extract used in the present invention can be obtained, for example, by extracting beetroot tea with an aqueous solvent. The raw material, beetroot tea, is a perennial shrub of the family Rosaceae, which has been used as sweet tea since ancient times in China. This beetroot tea can be subjected to extraction using leaves or stems, especially leaves dried in the sun, as a raw material.

The aqueous solvent used for extraction may be either water alone or an arbitrary mixed solution of water and one or two polar solvents such as lower alcohols such as methanol and ethanol and acetone. However, since the active ingredient of the present invention cannot be efficiently extracted only by the polar solvent, it is desirable that the active ingredient of the present invention is always mixed with water and the mixing ratio is 90% or less of the solvent. Among these solvents, water, ethanol, or a mixture thereof is preferably used from the viewpoint of safety, considering that the extract will be finally mixed in foods and the like.

The ratio of beet tea and solvent at the time of extraction is also not particularly limited, but one beet tea and a solvent 2-1.
000 times weight, especially 5-100 in terms of extraction operation and efficiency
Weight times are preferred. The extraction temperature is conveniently in the range of the boiling point of the solvent at room temperature and normal pressure, and the extraction time is preferably in the range of 10 minutes to 24 hours.

The soybean tea water-based solvent extract thus obtained as it is, concentrated product thereof, dried product obtained by removing solvent from the eluate, etc. can be used in any state, but the preservability is good. From the viewpoint of the safety of the organic solvent, it is preferable to use a dried product.

The medicine of the present invention is prepared by preparing the beetroot extract as it is or by formulating it with a known pharmaceutical carrier. The medicine of the present invention includes oral agents such as tablets, granules, powders, syrups; suppositories, parenteral agents such as external preparations, candy, chewing gum, milk, yogurt, whey beverages, lactic acid bacteria beverages, juices, It has a dosage form to be added to foods such as carbonated drinks, ice cream, puddings, water cans, etc., and is a dosage form to be added to cosmetics such as lotion, cosmetic cream, emulsion, foundation, lipstick, hair styling agent, hair tonic, hair restoration agent, etc. It may be in a dosage form to be blended with daily necessities such as toothpaste, toothpaste, mouthwash, shampoo, rinse and bath salt.

The pharmaceutical carrier that can be used in the preparation of the medicament of the present invention is not particularly limited, and those commonly used can be used, examples of which are:
Solid carriers such as starch, lactose, sucrose, mannitol, carboxymethyl cellulose, corn starch, and inorganic salts; distilled water, physiological saline, glucose aqueous solution, alcohols such as ethanol, liquid carriers such as propylene glycol, polyethylene glycol; various animal and vegetable oils , Oily carriers such as white petrolatum, paraffin and waxes.

In order to produce the above foods, cosmetics, toothpaste, shampoo and the like using the medicine of the present invention, appropriate components usually used can be added depending on the kind of the product. For example, when preparing food,
Glucose, fructose, sucrose, maltose, sorbitol,
Stevioside, rubusoside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L
-Ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, arabic gum, carrageenan, casein, gelatin , Pectin, agar, vitamin Bs, nicotinamide, calcium pantothenate, amino acids, calcium salts, pigments,
It can be produced by appropriately blending ingredients used as usual food materials such as flavors and preservatives.

Further, when preparing cosmetics, oils and fats such as vegetable oils, waxes such as lanolin and beeswax, hydrocarbons, fatty acids, higher alcohols, esters, various surfactants, pigments, fragrances, Vitamin, plant / animal extract component, ultraviolet absorber, antioxidant, antiseptic / bactericidal agent and the like, which are commonly used as raw materials for cosmetics, can be appropriately mixed and produced.

In the case of preparing cosmetics and the like using the beet extract of the present invention, other anti-inflammatory / anti-allergic cosmetic raw materials, for example, licorice extract component (particularly glycyrrhizic acid),
The effect can be enhanced by using diphenhydramine hydrochloride, azulene, dl-α-tocopherol and its derivative, vitamins B ₂ and B ₆ , and the like. Since beet extract strongly inhibits hyaluronidase, which is a hyaluronan degrading enzyme, its moisturizing effect can be further enhanced by using it together with hyaluronic acid. Even though bean tea extract alone has an indirect moisturizing and skin-refining effect by inhibiting the decomposition of hyaluronic acid in the skin, other moisturizing and skin-care cosmetic ingredients such as elastin, collagen, lecithin, squalene, placenta Liquid (placenta extract), glycerins, glycols, fermentation metabolites, lactic acid bacteria culture fluid, vitamins A and C, sodium chondroitin sulfate, sodium 2-pyrrolidone-5-carboxylate (PCA-Na), Bakumondou mucilage polysaccharides, etc. It is desirable to further enhance the effect by using it together with the plant polysaccharides of.

Since the ancient tea has been used as sweet tea in China since ancient times, the extract used in the present invention has no problem in terms of safety. However, it is desirable that the content of the beetroot extract in the medicine of the present invention is in the range of 0.01 to 5.0% in terms of dry weight in terms of effects and fragrance and color tone when added.

[0022]

[Effects and effects of the invention] The cyclooxygenase inhibitory activity of the tea extract, which is the active ingredient of the pharmaceutical composition of the present invention, that is, the anti-inflammatory activity due to the inhibition of PG combination is about 1/5 of that of aspirin, and the antihistamine activity is antiallergic. It was comparable to that of DSCG, the active ingredient of the drug intal.
Therefore, the drug of the present invention is useful as a drug having an anti-inflammatory effect and an anti-allergic effect. Also,
By daily ingestion of foods containing beet extract,
It can prevent and improve inflammation, sore throat, hay fever, and cough associated with a cold. Similarly, itching of the skin and the like can be ameliorated by using a cosmetic containing the tea extract.

The beet green tea extract according to the present invention is a potent inhibitor of hyaluronic acid decomposition and suppresses the decrease in the content of hyaluronic acid contained in the skin, arterial wall, joint cavity and the like. Although this is an indirect action, it enhances the moisture retention and flexibility of the skin, prevents arteriosclerosis associated with aging, and contributes to the improvement of arthritis.

[0024] The beet extract is a plant extract of which safety has been established, and has various effects as a hyaluronic acid decomposition inhibitor which acts as an anti-inflammatory agent and an antiallergic agent. Multifunctionality is unique.

[0025]

[Examples] Next, the present invention will be described with reference to Examples relating to a method for producing a tea extract, a cyclooxygenase inhibition test, a hyaluronidase inhibition test, an antihistamine test and an anti-inflammatory agent, an antiallergic agent, and a hyaluronic acid decomposition inhibitor. This will be described in more detail. Practical Example 1 Production of beetroot extract: 100 g of beetroot tea was placed in a 3000 ml Erlenmeyer flask, 1000 ml of hot water was added, and extraction was carried out in a boiling water bath for 3 hours. This was filtered, and the obtained filtrate was freeze-dried to obtain 32.3 g of an extract (golden tea extract 1).

Example 2 Preparation of beet tea extract: 100 g of beetroot tea was placed in a 3000 ml Erlenmeyer flask, and 1000 ml of 50% by volume ethanol was added.
1 was added, and the mixture was gently stirred at room temperature for 1 hour and extracted for 24 hours. This was filtered, and the obtained filtrate was concentrated under reduced pressure to remove ethanol, water was added, and the mixture was freeze-dried to obtain 31.1 g of an extract (golden tea extract 2).

Example 3 Assay of Cyclooxygenase Inhibitory Activity: The extract obtained in Example 1 was measured for its cyclooxygenase inhibitory activity by the following method. The results are shown in Table 1.

(Measurement method) The inhibitory effect on the cyclooxygenase activity of sheep seminal vesicle gland microsomes (Funakoshi Pharmaceutical, manufacturer code A3) was measured by the oxygen absorption method. That is, in a 4 ml reaction tank equipped with an enzyme electrode,
3.385 ml of 0.2 M Tris-hydrochloric acid buffer solution (pH 8.0) containing 10 mM L-tryptophan, 25 μl of 40 μM hemoglobin solution, 100 μl of test sample solution and 30 μl of the above buffer solution containing 0.6 mg of microsome were added, and 5
After preincubation for 10 minutes, the reaction was started by adding 10 μl of 10 mM arachidonic acid solution dissolved in ethanol. The reaction temperature is 30 ° C. The inhibitory rate I (%) was determined by the following calculation formula, where C is the initial enzyme absorption rate of the control in which purified water was added instead of the test sample solution and S was that when the test sample was added. I (%) = 100 × (C−S) / C In both the Chinese tea and the Rakan fruit extract, the dose-dependent curve of the inhibitory activity did not show linearity, and it became an S-shape with an inflection point near I = 50%. It was

[0029]

Example 4 Assay of Hyaluronidase Inhibitory Activity: The extract obtained in Example 1 was measured for its hyaluronidase inhibitory activity by the following method. The results are shown in Table 2.

(Measurement method) Hyaluronidase (Sigma, Type IV) derived from beef testis was used to measure mainly the inhibitory effect of compound 48/80 on the activation stage of the inactive enzyme. The enzymatic activity was measured by colorimetrically quantifying the increase in reducing power of the tetrasaccharide having N-acetylhexosamine, which is produced by hydrolysis of hyaluronic acid, as a reducing end by A ₅₆₅ (Maeda Yumie et al. ,
31: 233-237, 1990). That is, 100 μl of an appropriate amount of test sample was added in 0.1 M acetate buffer (pH 4.0).
0.10 mg of enzyme (100 NF units) dissolved in 50 μl of the same buffer was added, incubated at 37 ° C. for 20 minutes, and then compound 48/80 (5 dissolved in 100 μl of the same buffer).
0 μg), and further incubate at 37 ° C. for 20 minutes. Finally, hyaluronic acid sodium salt (200 µg, derived from a microorganism) dissolved in 250 µl of the same buffer was added, and the mixture was incubated at 37 ° C for 40 minutes, and then 0.4N.
After adding 100 μl of NaOH and ice-cooling, 100 μl of borate buffer (pH 9.1) was added and boiled for 3 minutes. After cooling with ice, p-dimethylaminobenzaldehyde reagent solution 3m
1 and incubated at 37 ° C. for 20 minutes, and then A
₅₆₅ was measured. As a control, the acetate buffer was used instead of the sample solution. Further, as each blank, the above-mentioned acetate buffer was used instead of the enzyme solution. The inhibitory activity was represented by the inhibition rate calculated from the following formula. A: Control solution A ₅₆₅ B: Control solution blank A ₅₆₅ C: Sample solution A ₅₆₅ D: Sample solution blank A ₅₆₅

[0032]

Example 5 Histamine release inhibition test from mast cells: The histamine release inhibitory activity of the extract obtained in Example 1 was measured by the following method (Yuko Hirai et al .: Jpn.
7, vol. 374-380, 1983). The results are shown in Table 3.

(Measurement Method) Bovine serum albumin (BS) was collected from peritoneal cells obtained by injecting Tyrode's solution into the abdominal cavity of exsanguinated Wistar rats.
A) -Mast cells were isolated by multi-layer centrifugation using physiological saline (specific gravity 1.068). The number of cells obtained is 2 x 10
^The cells were suspended in Tyrode's solution containing 0.1% BSA at ⁶ cells / ml to prepare a cell suspension. Sample solution 10 μl
10 μl of the cell suspension was added to the mixture and allowed to stand at 37 ° C. for 10 minutes, and then compound 48/80 as a degranulation inducer was added.
(5 μg / ml) (20 μl) was added, and the mixture was reacted at 37 ° C. for 10 minutes. Then, once ice-cooling, centrifuge (15
The amount of histamine released in the supernatant (0 × g, 5 minutes) was measured by high performance liquid chromatography with a fluorescence detector. The histamine release inhibitory activity was calculated by the following formula. Histamine release inhibition rate (%) = 100 x [1- (SR-
C) / (RC)] C: Amount of histamine released from control cells R: Amount of histamine released from cells when an inducer is added SR: When an inducer is added in the presence of a sample Amount of histamine released from cells

[0035]

Example 6 Tablets: 150 g of beetroot extract 1 was mixed with the same amount of lactose and 5 g of magnesium stearate, and the mixture was tabletted with a single-shot tableting machine to give a diameter of 10 mm and a weight of 300 mg.
Tablets were produced.

Example 7 Condyle Granules: The tablets obtained in Example 6 were crushed, sized and sieved to obtain 20-50 mesh granules.

Example 8 candy: (composition) (weight part) powdered sorbitol 99.7 fragrance 0.2 beet tea extract 0.05 sorbitol seed 0.05 total amount 100

Example 9 Candy: (composition) (weight part) sand sugar 47.0 water candy 49.76 flavor 1.0 water 2.0 beet extract 0.24 total 100

Example 10 Lozenge: (composition) (weight part) gum arabic 6 glucose 73 sugar beet extract 0.05 dipotassium phosphate 0.2 0.2 potassium diphosphate 0.1 lactose 17 flavor 0.1 Magnesium stearate Residual total 100

Example 11 Gum: (composition) (weight part) Gum base 20 Calcium carbonate 2 Stevioside 0.1 Beet tea extract 0.05 Lactose 76.85 Flavor 1 Total amount 100

Example 12 Chewing gum: (composition) (weight part) gum base 20.0 sand sugar 75.62 flavoring agent 1.0 citric acid 1.0 water 2.0 beet extract 0.38 total amount 100

Example 13 Caramel: (composition) (weight part) Granulated sugar 32.0 Water candy 20.0 Powdered milk 40.0 Hardening oil 4.0 Food salt 0.6 Perfume 0.02 Water 3.22 Tencha extract 0.16 Total 100

Example 14 Coffee jelly: (composition) (weight part) Granulated sugar 15.0 Gelatin 1.0 Coffee extract 5.0 Water 78.93 Beet tea extract 0.07 Total amount 100

Example 15 Ice cream: (composition) (weight part) fresh cream (45% fat) 33.8 nonfat dry milk 11.0 granulated sugar 14.8 sweetened egg yolk 0.3 vanilla essence 0.1 water 39.93 Beetle tea extract 0.07 Total amount 100

Example 16 Custard pudding: (composition) (weight part) milk 47.51 whole egg 31.9 top white sugar 17.1 water 3.4 sugar beet extract 0.09 total quantity 100

Example 17 Water yokan: (composition) (weight part) red raw bean jam 24.8 powdered agar 0.3 food salt 0.1 top white sugar 24.9 sugar beet extract 0.1 water 49. 8 total 100

Example 18 Juice: (composition) (weight part) Frozen concentrated Unshu mandarin orange juice 5 Fructose dextrose liquid sugar 11 Citric acid 0.2 L-ascorbic acid 0.02 Chinese tea extract 0.05 Perfume 0 .2 Chromium 0.1 Water remaining Total 100

Example 19 Carbonated beverage: (composition) (weight part) granulated sugar 8.0 concentrated lemon juice 1.0 L-ascorbic acid 0.10 citric acid 0.06 sodium citrate 0.05 coloration Material 0.05 Fragrance 0.15 Carbonic acid water 90.55 Beet tea extract 0.04 Total amount 100

Example 20 Lactobacillus beverage: (composition) (weight part) 21% milk solids fermented milk 14.76 fructose glucose liquid sugar 13.31 pectin 0.5 quinic acid 0.08 Fragrance 0.15 Water 71.14 Horse chestnut extract 0.06 Total amount 100

Example 21 Coffee Beverage: (Composition) (Part by Weight) Granulated Sugar 8.0 Degreased Powdered Milk 5.0 Caramel 0.2 Coffee Extract 2.0 Flavor 0.1 Poly Glycerin 0.05 Fatty acid ester Food salt 0.05 Water 84.56 Beet tea extract 0.04 Total amount 100

Example 22 Dentifrice: (Composition) (Part by weight) Dibasic calcium phosphate 42 Glycerin 18 Carrageenan 0.9 Sodium lauryl sulfate 1.2 Sodium saccharin 0.09 Butyl paraoxybenzoate 0.005 Horse chestnut extract 0 .05 Fragrance 1 Water Remaining Total 100

Example 23 Mouthwash: (Composition) (Parts by weight) Sodium lauryl sulfate 0.8 Glycerin 7 Sorbitol 5 Ethyl alcohol 15 Horse chestnut extract 0.05 1-Menthol 0.05 Perfume 0.04 Saccharin sodium 0.1 Water Residual total 100

Example 24 Soft lotion (weakly acidic): (composition) (weight part) glycerin 5.0 propylene glycol 4.0 sodium hyaluronate 0.1 wart tea extract 0.05 polyoxyethylene sorbitan monolaurin Acid ester 1.5 (20E.O.) Polyoxyethylene lauryl 0.5 Ether (20E.O.) Ethanol 10.0 Fragrance 0.1 Dye suitable amount Preservative suitable amount UV absorber suitable amount refined Water 78.75

Example 25 Emollient cream: (composition) (part by weight) beeswax 2.0 stearyl alcohol 5.0 stearic acid 8.0 squalane 10.0 self-emulsifying propylene glycol 3.0 monostearate polyoxy Ethylene cetyl ether 1.0 (20 EO) Fragrance 0.5 Preservative proper amount Antioxidant proper amount Propylene glycol 7.8 Glycerin 4.0 Sodium hyaluronate 0.1 Beet extract 0.1 Triethanolamine 1.0 Purified water 57.5

Example 26 Emollient lotion: (composition) (weight part) stearic acid 2.0 cetanol 1.5 vaseline 3.0 lanolin alcohol 2.0 liquid paraffin 10.0 polyoxyethylene monooleic acid 2. 0 Ester (10 EO) Fragrance 0.5 Antioxidant Appropriate amount Preservative Appropriate amount Propylene Glycourt 4.8 Glycerin 3.0 Sodium hyaluronate 0.1 Beet extract 0.1 Triethanolamine 1. 0 refined water 70.0

Example 27 Milky liquid foundation: (composition) (weight part) stearic acid 2.4 propylene monostearate 2.0 glycol glycol cetostearyl alcohol 0.2 liquid lanolin 2.0 liquid paraffin 3.0 myristin Isopropyl acid 8.5 Propyl paraoxybenzoate Appropriate amount Purified water 64.1 Sodium carboxymethyl cellulose 0.2 Bentonite 0.5 Propylene glycol 3.8 Sodium hyaluronate 0.1 Beet tea extract 0.1 Triethanolamine 1.1 Methyl paraoxybenzoate Appropriate amount Titanium oxide 8.0 Tark 4.0 Coloring pigment Appropriate amount Fragrance Appropriate amount

Example 28 Hair-growth hair tonic: (composition) (weight part) ethanol 70.0 assembly extract 0.2 vitamin E 0.2 L-menthol 0.4 glycyrrhizin 0.1 beetroot extract 0.1 Salicylic acid 0.5 Glycerin 0.5 Perfume Appropriate amount Purified water 28.0

Example 29 Shampoo: (Composition) (Part by weight) Alkyl ether sulfate 16.0 Sodium (AES-Na) Lauric acid diethanolamide 4.0 Propylene glycol 1.9 Beet tea extract 0.1 Preservative , Dye, Fragrance, Proper amount, Purified water 78.0

Example 30 Rinse: (composition) (weight part) stearyl dimethyl chloride 1.4 benzylammonium stearyl alcohol 0.6 glycerin monostearate 1.5 diet salt 0.1 beet extract 0.1 sperm Water production 96.3 or above

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[Procedure amendment]

[Submission date] March 15, 1991

[Procedure Amendment 1]

[Document name to be amended] Statement

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[0002]

2. Description of the Related Art With the development of science and technology, many compounds have been synthesized and drugs having various actions have been found.
For example, a large number of drugs, mainly chemically synthesized products, have been developed as anti-inflammatory agents. And, about 100 years ago, the point of action of aspirin, which has been clinically used, is the inhibition of cyclooxygenase, which is the initial enzyme in the process from arachidonic acid to the synthesis of prostaglandins (hereinafter abbreviated as "PG"). With the elucidation of the above, a drug having a strong medicinal effect by a similar mechanism of action, such as indomethacin, has been developed.

[Procedure Amendment 2]

[Document name to be amended] Statement

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As an antiallergic agent, a chemically synthesized product such as disodium cromoglycate (hereinafter abbreviated as "DSCG"), and caffeic acid as an inhibitor of 5-lipoxygenase that produces leukotriene from arachidonic acid. Mugwort extract and the like, which are contained in large amounts, are used.

[Procedure 3]

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On the other hand, hyaluronic acid derived from microorganisms is mainly mixed in cosmetics as a moisturizing agent, and recently, the inhibitory activity against hyaluronidase, which is a hyaluronic acid-degrading enzyme, has been used as an index for the anti-tea extract. Attempts have been made to evaluate allergic activity (Yumie Maeda et al .: Shokuei, Vol. 31, pp. 233-237, 199).
0), a correlation between hyaluronidase inhibitory activity and antiallergic activity is suggested.

[Procedure amendment 4]

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[0009]

Means for Solving the Problems To solve the above problems, the present inventor solves the above problems by using both cyclooxygenase, which is the initial enzyme in the process of PG synthesis from arachidonic acid, and hyaluronidase, which is a hydrolase of hyaluronic acid. We conducted an intensive search to find a natural product that inhibits the above and also has antihistamine activity. Here, the purpose of inhibiting hyaluronidase is that inhibition of this enzyme strongly correlates not only with maintaining the level of hyaluronic acid in the human body, but also with anti-inflammatory and anti-allergic activities. This enzyme is thought to be activated during inflammation, destroy the connective tissue matrix, and enhance the permeability of inflammatory cells and blood vessels, so inhibition of this enzyme may lead to anti-inflammation. To be done. In addition, hyaluronidase is likely to be involved in a series of processes from the binding of IgE-antigen complex to the receptor in mast cells to the release (degranulation) of histamine granules. This is because a large morphological change such as degranulation cannot occur without temporary destruction of the water retention structure by hyaluronic acid. In fact, many anti-allergic agents also have hyaluronidase inhibitory activity.

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The ratio of beet tea and solvent at the time of extraction is also not particularly limited, but one beet tea and a solvent 2-1.
000 times by weight, particularly 5-100 times by weight in terms of extraction operation and efficiency. The extraction temperature is conveniently in the range of room temperature-boiling point of the solvent under normal pressure, and the extraction time is preferably in the range of 10 minutes to 24 hours.

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[0022]

[Effects and effects of the invention] The cyclooxygenase inhibitory activity of the tea extract, which is the active ingredient of the medicament of the present invention, that is, the anti-inflammatory activity due to the inhibition of PG synthesis is about 1/5 of that of aspirin, and the antihistamine activity is the antiallergic activity. It was comparable to that of DSCG, the active ingredient of the drug intal.
Therefore, the drug of the present invention is useful as a drug having an anti-inflammatory effect and an anti-allergic effect. In addition, by regularly ingesting foods containing the tea extract, it is possible to prevent inflammation, throat swelling, hay fever, cough, etc. associated with colds.
Can be improved. Similarly, itching of the skin and the like can be ameliorated by using a cosmetic containing the tea extract.

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[0024] Beet extract is a plant extract of which safety has been established, and has various effects as a hyaluronic acid decomposition inhibitor in addition to its action as an anti-inflammatory agent and antiallergic agent. , Its versatility is unique.

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[0025]

[Examples] Next, examples of a method for producing a tea extract, a cyclooxygenase inhibition test, a hyaluronidase inhibition test, a histamine release inhibition test, and an anti-inflammatory agent, an antiallergic agent, and a hyaluronic acid decomposition inhibitor, will be given.
The present invention will be described in more detail. Example 1 Preparation of beet tea extract: 100 g of beetroot tea was placed in a 3000 ml Erlenmeyer flask, 1000 ml of hot water was added, and extraction was performed in a boiling water bath for 3 hours. This was filtered, and the obtained filtrate was freeze-dried to obtain 32.3 g of an extract (golden tea extract 1).

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(Measurement method) The inhibitory effect on the cyclooxygenase activity of sheep seminal vesicle microsomes (Funakoshi Pharmaceutical, maker code A3) was measured by the oxygen absorption method. That is, 10 m in a 4 ml reaction tank equipped with an oxygen electrode.
3.835 ml of 0.2 M Tris-hydrochloric acid buffer solution (PH8.0) containing M L-tryptophan, 25 μl of 40 μM hemoglobin solution, 100 μl of test sample solution and 30 μl of the above-mentioned buffer solution containing 0.6 mg of microsomes were added, and pre-treated for 5 minutes. After incubation 10 dissolved in ethanol
The reaction was started by adding 10 μl of mM arachidonic acid solution. The reaction temperature is 30 ° C. Letting C be the initial enzyme absorption rate of the control in which purified water was added instead of the test sample solution, and S be the time when the test sample was added, the inhibition rate I was calculated by the following formula.
(%) Was calculated. I (%) = 100 × (C−S) / C In both the Chinese tea and the Rakan fruit extract, the dose-dependent curve of the inhibitory activity did not show linearity, and it became an S-shape with an inflection point near I = 50%. It was

[Procedure Amendment 10]

[Document name to be amended] Statement

[Name of item to be amended] Detailed explanation of the invention

[Correction method] Change

[Correction content]

(Measurement method) Using a hyaluronidase derived from beef testis (Sigma, Type IV), the inhibitory effect of compound 48/80 on the activation stage of the inactive enzyme was measured. Enzymatic activity, by the increase in reducing power of tetrasaccharide to A ₅₈₅ Colorimetric that the N- acetylhexosamine produced by hydrolysis of hyaluronic acid and the reducing end was determined (Yumi Maeda Megumira: ShokuMamorushi, 31
Vol. 233-237, 1990). That is, an appropriate amount of test sample was added to 100 μM of 0.1 M acetate buffer (pH 4.0).
0.10 mg of enzyme (100 NF units) dissolved in 50 μl of the same buffer was added, incubated at 37 ° C. for 20 minutes, and then compound 48/80 (50) dissolved in 100 μl of the same buffer.
μg), and further incubate at 37 ° C. for 20 minutes. Finally, add hyaluronic acid sodium salt (200 μg, derived from microorganism) dissolved in 250 μl of the same buffer,
After incubating at 37 ℃ for 40 minutes, 0.4NNa
100 μl of OH was added and ice-cooled, and then borate buffer (pH 9.
1) Add 100 μl and boil for 3 minutes. After ice cooling, p-
Add 3 ml of dimethylaminobenzaldehyde test solution,
After incubating at 37 ° C. for 20 minutes, A ₅₈₅ was measured. As a control, the acetate buffer was used instead of the sample solution. In addition, as each blank, the above-mentioned acetate buffer was used instead of the enzyme solution. The inhibitory activity was represented by the inhibition rate calculated from the following formula.

[Equation 1] A: Control solution A ₅₈₅ B: Control solution blank A ₅₈₅ C: Sample solution A ₅₈₅ D: Sample solution blank A ₅₈₅

[Procedure Amendment 11]

[Document name to be amended] Statement

[Name of item to be amended] Detailed explanation of the invention

[Correction method] Change

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Example 5 Test for inhibiting histamine release from mast cells: The histamine release inhibiting activity of the extract obtained in Example 1 was measured by the following method (Yuko Hirai et al .: Jpn.
Vol. 374-380, 1983). The results are shown in Table 3. ─────────────────────────────────────────────────── ───

[Procedure amendment]

[Submission date] March 8, 1994

[Procedure Amendment 1]

[Document name to be amended] Statement

[Name of item to be corrected] 0002

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[0002]

2. Description of the Related Art With the development of science and technology, many compounds have been synthesized and drugs having various actions have been found.
For example, a large number of drugs, mainly chemically synthesized products, have been developed as anti-inflammatory agents. And, about 100 years ago, the point of action of aspirin, which has been clinically used , is the inhibition of cyclooxygenase, which is the initial enzyme in the process from arachidonic acid to the synthesis of prostaglandins (hereinafter abbreviated as "PG"). With the elucidation of the above, a drug having a strong medicinal effect by a similar mechanism of action, such as indomethacin, has been developed.

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[Name of item to be corrected] 0003

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As an antiallergic agent, a chemically synthesized product such as disodium cromoglycate (hereinafter abbreviated as "DSCG"), and caffeic acid as an inhibitor of 5-lipoxygenase that produces leukotriene from arachidonic acid. Mugwort extract and the like, which are contained in large amounts, are used.

[Procedure 3]

[Document name to be amended] Statement

[Name of item to be corrected] 0005

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On the other hand, hyaluronic acid derived from microorganisms is mainly mixed in cosmetics as a moisturizing agent, and recently, the inhibitory activity against hyaluronidase, which is a hyaluronic acid-degrading enzyme, has been used as an index for the anti- tea extract. Allele
Attempts have been made to evaluate ghee activity (Yumie Maeda et al .: Shokuei, Vol. 31, pp. 233-237, 199).
0), a correlation between hyaluronidase inhibitory activity and antiallergic activity is suggested.

[Procedure amendment 4]

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[Correction target item name] 0009

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[0009]

[Means for Solving the Problems] In order to solve the above problems, the present inventor has selected both enzymes, cyclooxygenase, which is the initial enzyme in the process leading to the synthesis of PG from arachidonic acid, and hyaluronidase, which is a hydrolase of hyaluronic acid. An intensive search was conducted to find a natural product that inhibits and also has antihistamine activity. Here, the purpose of inhibiting hyaluronidase is that inhibition of this enzyme strongly correlates not only with maintaining the level of hyaluronic acid in the human body, but also with anti-inflammatory and anti-allergic activities. This enzyme is thought to be activated during inflammation, destroy the connective tissue matrix, and enhance the permeability of inflammatory cells and blood vessels, so inhibition of this enzyme may lead to anti-inflammation. To be done. In addition, hyaluronidase is likely to be involved in a series of processes in mast cells, from the binding of IgE-antigen complex to the receptor to the release (degranulation) of histamine granules. This is because a large morphological change such as degranulation cannot occur without temporary destruction of the water retention structure by hyaluronic acid. In fact, anti-allele
Many ghee agents also have hyaluronidase inhibitory activity.

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[Correction target item name] 0014

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The ratio of beet tea and solvent at the time of extraction is also not particularly limited, but one beet tea and a solvent 2-1.
000 times by weight, particularly 5-100 times by weight in terms of extraction operation and efficiency. Extraction temperature room - is conveniently in the range of the boiling point of the solvent under normal pressure, the extraction time is preferably in the range of 10 minutes to 24 hours.

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[Document name to be amended] Statement

[Name of item to be corrected] 0022

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[0022]

ACTION AND EFFECTS OF THE INVENTION The cyclooxygenase inhibitory activity of the tea extract, which is the active ingredient of the pharmaceutical composition of the present invention, that is, the anti-inflammatory activity due to the inhibition of PG synthesis is about 1/5 of that of aspirin, and the antihistamine activity is the antiallergic agent. It was comparable to that of DSCG, the active ingredient of Intar. Therefore, the drug of the present invention is useful as a drug having an anti-inflammatory effect and an anti-allergic effect. Further, by ingesting foods containing the extract of beetroot tea on a daily basis, it is possible to prevent and improve inflammation, sore throat, hay fever, cough and the like associated with a cold. Similarly, itching of the skin and the like can be ameliorated by using a cosmetic containing the tea extract.

[Procedure Amendment 7]

[Document name to be amended] Statement

[Name of item to be corrected] 0024

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[0024] Beet extract is a plant extract of which safety has been established, and has various effects as a hyaluronic acid decomposition inhibitor in addition to its action as an anti-inflammatory agent and antiallergic agent. , Its versatility is unique.

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[Document name to be amended] Statement

[Name of item to be corrected] 0025

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[0025]

[Examples] Next, a method for producing beet tea extract, cyclooxygenase inhibition test, hyaluronidase inhibition test, histamine
Examples relating to release inhibition test and production of anti-inflammatory agent, anti-allergic agent, and inhibitor for degradation of hyaluronic acid,
The present invention will be described in more detail. Example 1 Production of beet tea extract: 100 g of beetroot tea was placed in a 3000 ml Erlenmeyer flask, 1000 ml of hot water was added, and extraction was carried out in a boiling water bath for 3 hours. This was filtered, and the obtained filtrate was freeze-dried to obtain 32.3 g of an extract (golden tea extract 1).

[Procedure Amendment 9]

[Document name to be amended] Statement

[Correction target item name] 0028

[Correction method] Change

[Correction content]

(Measurement method) The inhibitory effect on the cyclooxygenase activity of sheep seminal vesicle microsomes (Funakoshi Pharmaceutical, maker code A3) was measured by the oxygen absorption method. That is, 10 m in a 4 ml reaction tank equipped with an oxygen electrode.
0.235M Tris-hydrochloric acid buffer solution (pH 8.0) containing 3.835 ml of M-L-tryptophan, 25 µl of 40 µM hemoglobin solution, 100 µl of test sample solution and 30 µl of the above-mentioned buffer solution containing 0.6 mg of microsomes were added, and pre-treated for 5 minutes. After incubation 10 dissolved in ethanol
The reaction was started by adding 10 μl of mM arachidonic acid solution. The reaction temperature is 30 ° C. Letting C be the initial enzyme absorption rate of the control in which purified water was added instead of the test sample solution, and S be the time when the test sample was added, the inhibition rate I was calculated by the following formula.
(%) Was calculated. I (%) = 100 × (C−S) / C In both the Chinese tea and the Rakan fruit extract, the dose-dependent curve of the inhibitory activity did not show linearity, and it became an S-shape with an inflection point near I = 50%. It was

[Procedure Amendment 10]

[Document name to be amended] Statement

[Correction target item name] 0031

[Correction method] Change

[Correction content]

(Measurement method) Using a hyaluronidase derived from beef testis (Sigma, Type IV), the inhibitory effect of compound 48/80 on the activation stage of the inactive enzyme was measured. Enzyme activity, an increase in the reducing power of tetrasaccharide to the N- acetylhexosamine produced by hydrolysis of hyaluronic acid and the reducing end by colorimetry at A _585, was determined (Yumi Maeda Megumira: ShokuMamorushi , 31
Vol. 233-237, 1990). That is, an appropriate amount of test sample was added to 100 M acetate buffer (pH 4.0) 100
0.10 mg of enzyme (100 NF units) dissolved in 50 μl of the same buffer was added, incubated at 37 ° C. for 20 minutes, and then compound 48/80 (50) dissolved in 100 μl of the same buffer.
μg), and further incubate at 37 ° C. for 20 minutes. Finally, add hyaluronic acid sodium salt (200 μg, derived from microorganism) dissolved in 250 μl of the same buffer,
After incubating at 37 ° C. for 40 minutes, 0.4 N N
After adding 100 μl of aOH and cooling with ice, borate buffer (pH
9.1) Add 100 μl and boil for 3 minutes. After ice cooling,
After adding 3 ml of a p-dimethylaminobenzaldehyde reagent solution and incubating at 37 ° C. for 20 minutes, A ₅₈₅ was measured. As a control, the acetate buffer was used instead of the sample solution. Further, as each blank, the above-mentioned acetate buffer was used instead of the enzyme solution. The inhibitory activity was represented by the inhibition rate calculated from the following formula. A: Control solution A ₅₈₅ B: Control solution blank A ₅₈₅ C: Sample solution A ₅₈₅ D: Sample solution blank A ₅₈₅

[Procedure Amendment 11]

[Document name to be amended] Statement

[Correction target item name] 0033

[Correction method] Change

[Correction content]

Example 5 Histamine Release Inhibition Test from Mast Cells: The histamine release inhibitory activity of the extract obtained in Example 1 was measured by the following method (Yuko Hirai et al .: Jpn.
Vol. 374-380, 1983). The results are shown in Table 3.

Claims (3)

[Claims] 1. A medicine selected from an anti-inflammatory agent, an anti-allergic agent and a hyaluronic acid decomposition inhibitor, which comprises an extract of beetroot tea as an active ingredient. 2. A cosmetic containing the medicine according to claim 1. 3. A food containing the medicine according to claim 1.