JPH0499712A - Production of anticarious agent and anticarious substance - Google Patents

Abstract

PURPOSE:To obtain an anticarious agent, containing an extract separated from Hydrangea aspera D. Don as an active ingredient and having inhibitory activity against glucosyltransferase. CONSTITUTION:Leaves or stems, especially the leaves of Hydrangea aspera D. Don which is a perennial shrub belonging to the family Saxifragaceae of the order Rosales are dried in the sun, used as a raw material and extracted with a mixed solvent of water and a lower alcohol such as methanol or ethanol or acetone at ambient temperature to the boiling point of the solvent under ordinary pressure. The resultant extract solution in an amount of 0.00001-0.5wt.% is contained as an active ingredient and further blended with suitable ingredients normally used according to the kind thereof and prepared to afford the objective substance. The aforementioned agent can be prepared as an oral agent such as a dentifrice, a mouthwash or a troche or in a dosage form added to foods such as sponge cakes, sweet jellies of beans, cakes, cookies, beverages, ice creams or caramels.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a method for producing an anti-caries agent and an anti-caries substance, and more specifically, the present invention relates to a method for producing an anti-caries agent and an anti-caries substance, and more specifically, the present invention relates to a method for producing an anti-caries agent and an anti-caries substance, and more particularly, the present invention relates to a method for producing an anti-caries agent and an anti-caries substance. The present invention relates to an anti-caries agent and a method for producing an anti-caries substance from sweet tea.

[Prior Art] Various hypotheses have been proposed regarding the cause of caries in the past, but it is now recognized that caries is a type of bacterial infection based on Miller's chemical bacteria theory.

The caries development mechanism based on this theory is as follows.

That is, oral streptococci, especially Streptococcus mutans (5trept).

An enzyme called glucosyltransferase produced by C. coccus mutans first uses sucrose in the mouth as a substrate to produce a sticky, insoluble polysaccharide (glucan). Streptococcus mutans (hereinafter referred to as r
3. Streptococcus mutans" bacteria adhere to the tooth surface and form dental plaque using the glucans they produce. In this dental plaque, various microorganisms including S. mutans coexist and proliferate, and the metabolism of these microorganisms produces organic acids, and the action of these organic acids lowers the pH of the tooth surface. Demineralization occurs on the enamel surface, causing caries and progressing.

In addition to dental caries, plaque formation is also thought to be a cause of periodontal disease and bad breath.

[Problem to be solved by the invention] Based on this knowledge, as a method for preventing caries, antibacterial agents for oral microorganisms, inhibitors of the above-mentioned glucosyltransferases, and polysaccharides formed by glucosyltransferases using sucrose as a substrate have been proposed. Various studies have been conducted on enzymes that degrade . However, no effective countermeasure has yet been found.

In particular, since dental plaque formed by oral streptococci, mainly S. mutans, is the cause of caries, it is possible to suppress the formation of plaque by inhibiting the glucosyltransferase activity of S. mutans. Although it is believed that glucosyltransferase activity can be an effective means for preventing the occurrence of cavities, at present no substance that inhibits glucosyltransferase activity that can be used in practice has been found.

[Means for Solving the Problems] The present inventors conducted intensive research to find a substance that effectively inhibits glucosyltransferase and does not have harmful effects on the human body, and as a result, the present inventors developed a sweet tea water system. The present invention has been completed based on the discovery that a solvent extract contains a substance that very effectively inhibits the activity of glucosyltransferase produced by S. mutans.

That is, an object of the present invention is to provide an anti-caries agent containing an aqueous solvent extract from light brown tea as an active ingredient.

The usucha used in the present invention is a perennial rose bush that has been used as sweet tea in China since ancient times.

This light tea can be made from the leaves or stems, especially the leaves, dried in the sun and subjected to extraction.

The aqueous solvent used for extraction may be water alone or any mixture of water and one or two polar solvents such as methanol, lower alcohols such as ethanol, and acetone.

However, since the active ingredient of the present invention cannot be efficiently extracted using polar solvents, it is preferable that the mixture is always mixed with water and that the mixing ratio of the solvent is 90% by volume or less.

Among these solvents, water, water,
Preferably, ethanol or a mixture thereof is used.

The ratio of light brown tea to solvent during extraction is not particularly limited either, but it is preferably 2 to 1000 times the weight of the solvent to 1 weight of light brown tea, and particularly 5 to 100 times the weight of the solvent in terms of extraction operation and efficiency.

The extraction temperature is conveniently within the range of room temperature to the boiling point of the solvent under normal pressure, and the extraction time is preferably within the range of 10 minutes to 24 hours.

Since the thus obtained sweet tea aqueous solvent extract itself has glucosyltransferase inhibitory activity, it can be used as it is as an anti-caries agent, but it can also be further subjected to adsorption treatment and the resulting adsorbed product can be used. Also good.

Adsorption treatment using a synthetic adsorbent is performed by passing the sweet tea water-based solvent extract through a column packed with a synthetic adsorbent, and both fractions that pass through the column are obtained as non-adsorbed fractions. On the other hand, the adsorbed fraction adsorbed on the column can be obtained by removing the non-adsorbed fraction by washing with water and then desorbing it with a solvent.

As the desorption solvent, a polar organic solvent such as aqueous ethanol can be used, but in order to increase the purity of the active ingredient, the concentration of the organic solvent is preferably such that the sweet component of the tea cloth is not desorbed from the resin.

The synthetic adsorbents used for the separation treatment of light brown aqueous solvent extracts are aromatic synthetic adsorbents manufactured by polymerizing methyl chloride and divinylbenzene, and methacrylic synthetic adsorbents manufactured by polymerizing methacrylic acid esters. As an adsorbent, the commercially available synthetic aromatic adsorbent is Diaion HP20.
, HP21 (Mitsubishi Chemical Industries), Amberlight XAD
2. XAD4 (Rohm & Haas, USA), etc.
Among methacrylic synthetic adsorbents, Sepabead HPIλ (G,
HP2MG (Mitsubishi Chemical Industries), Amberlight XAD
7, XAD8 (Rohm and Haas, USA)
etc. can be mentioned.

When treating an aqueous solvent extract of light brown with these synthetic adsorbents, in order to complete the fractionation, the organic solvent in the extract must be removed in advance by vacuum concentration, etc., or the extract must be thoroughly diluted with water, etc. Preferably, the treatment is carried out.

The sweet tea water-based solvent extract and the synthetic adsorbent-unadsorbed fraction and adsorbed fraction obtained as described above can be extracted or treated with a synthetic adsorbent as is, concentrated, or eluted with the solvent removed. Although it can be used in any state, such as a dried product that has been removed, it is preferable to use a dried product in terms of storage stability and safety of organic solvents.

The anti-caries agent of the present invention is prepared by blending the above-mentioned aqueous solvent extract, non-adsorbed fraction, adsorbed fraction, and mixtures thereof with various conventionally used components.

Preferred dosage forms of the anti-caries agent of the present invention include oral preparations such as toothpaste, mouthwash, and troches;
Castella, mizu yokan, dorayaki skin, sponge cake, butter cake, bavarois, custard cream, butter cream, custard pudding, kutsky, sweet bread, steamed bread, jam, lactic acid bacteria drink, carbonated drink, coffee drink, coffee jelly, caramel Examples include dosage forms added to foods such as ice cream, chewing gum, juice, candy, and chocolate.

In the production of these oral preparations and foods, appropriate commonly used ingredients can be used depending on their type. For example, for oral preparations, calcium carbonate, calcium diphosphate, silicic anhydride, magnesium carbonate, glycerin , sorbitol, propylene glycol, polyethylene glycol, carboxymethylcellulose, methylcellulose,
Sodium alginate, carrageenan, carboxyvinyl polymer dioctyl sodium sulfosuccinate, sodium lauryl sulfate, sodium dodecylbenzenesulfonate,
Butyl paraoxybenzoate, hinokitiol, allantoin, glycyrrhizin, alcohol, gum arabic, starch, corn starch, sodium saccharin, stevioside, lupusoside, glucose, lactose, magnesium stearate, potassium phosphate, dipotassium phosphate, menthol, eucalyptus oil , pelapermint, spearmint, pigments, etc., as well as fluorides such as sodium fluoride and sodium monofluorophosphate, lysozyme chloride,
Anti-inflammatory agents such as azulene, sodium chloride, etc. can be appropriately blended. Food products include glucose, fructose, sucrose, maltose, sorbitol, stevioside, rubusoside, corn syrup, lactose, citric acid, tartaric acid,
Malic acid, succinic acid, )'L 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, gum arabic, carrageenan, casein, gelatin, pectin, agar , vitamin B
It can be produced by suitably blending those commonly used as food raw materials, such as nicotinamide, calcium pantothenate, amino acids, calcium salts, pigments, fragrances, and preservatives.

Sweet tea has been used as sweet tea in China since ancient times, and the extract used in the present invention, as well as the non-adsorbed fraction and adsorbed fraction obtained by treating it with a synthetic adsorbent, have no safety issues. do not have. However, the blending amount of the active ingredient in the anti-caries agent of the present invention is calculated from 0.00001 to 0.000 on dry weight basis based on the anti-caries activity, aroma and color tone when added.
．． Preferably, the temperature range is 5%.

[Actions and Effects of the Invention] Catechins have been known as compounds that have a growth-inhibiting effect on S. mutans, and a plant containing a large amount of catechins is the tea plant.

However, the activity of catechins to inhibit the growth of S. mutans is extremely weak, and from the viewpoint of taste and smell, almost no anti-caries activity can be expected at practically usable concentrations.

On the other hand, since sweet tea is a perennial shrub of the Rosaceae family and its extract has glucosyltransferase inhibitory activity in extremely small amounts, it is clear that the effects of the present invention are not related to catechins.

The anti-inflammatory agent of the present invention described above is a new type of anti-inflammatory agent that utilizes glucosyltransferase inhibitory activity.
It can be used to prevent caries, etc.

[Example] Next, examples related to a method for producing a light tea extract and a non-adsorbed fraction and an adsorbed fraction obtained by treating it with a synthetic adsorbent, a glucosyltransferase inhibitory activity test, and the production of an anti-fluxing agent will be given, and will demonstrate the present invention. will be explained in more detail.

Example 1 Production of light tea extract: 100 g of sweet tea was placed in a 3000 ml Erlenmeyer flask, 10100 O of hot water was added, and extraction was performed in a boiling water bath for 3 hours. This was filtered, and the resulting moat was freeze-dried.1. extract 3
2.3g was obtained.

Example 2 Production of light tea extract: Put 100g of sweet tea into a 3000ml Erlenmeyer flask,
Add 1000ml of 0% ethanol, and at room temperature,
Extraction was performed for 24 hours with gentle stirring every hour. This was filtered, and the resulting liquid was drifted under reduced pressure to remove ethanol, then water was added and freeze-dried to obtain 1 g of extract 3].

Example 3 Adsorption treatment of light tea extract 3.2 g of extract from 10 g of sweet tea obtained according to the method of Example 1 was dissolved in 200 ml of water, and this was poured into a column (11 x 210 mm) packed with Diaion HP20. It was adsorbed. Both fractions that were not adsorbed to the adsorbent and the washed fraction washed with 200 ml of water after the adsorption operation were combined to form a non-adsorbed fraction, which was lyophilized to obtain 1.7 g of a sample.

The adsorbed M fraction was sequentially desorbed with 20% ethanol (5 times the volume), 30% ethanol (5 times the volume), and 50% ethanol (10 times the volume), concentrated under reduced pressure, and then lyophilized by adding water. Samples weighing 0.47 g, 0.25 g, and 0.67 g were obtained, respectively.

Of these, the adsorbed fraction desorbed with 50% ethanol is
In particular, it contained a large amount of sweet components, and the glucosyltransferase inhibitory activity at a concentration of ioooppm was 43%.

Example 4 Assay of glucosyltransferase inhibitory activity: The glucosyltransferase inhibitory activity of the extracts obtained in Examples 1 and 2 and the non-adsorbed fraction and adsorbed M fraction of the adsorption treatment of Example 3 was measured by the following method. . The results are shown in Table 1.

(Measurement method) 0.6 ml of 500 mK mirin sodium mild WE solution (pH 6.0) containing 5% sucrose, 0.5%, dextran T10, and 0.5% sodium azide, 20
A reaction system was prepared by adding 0.15 ml of an aqueous solution of the sample prepared to 0 ppm, an enzyme solution prepared from S. mutans, and water in a total amount of 3 ml, and the reaction was carried out in a glass test tube. The enzyme solution was prepared by culturing S. mutans MT8148 strain in Todd-1ewitt medium and extracting it from the bacterial cells with 8M urea (
S., Hamada et al., J., Gen., M.
icrobiol, volume 135, pages 335-344 (
1,989)). In addition, the amount of enzyme was set so that the absorbance at 550 nm was approximately 1.0 after a 13-hour reaction at 37°C.

The produced insoluble glucan was disrupted by ultrasonication, and the absorbance (A) at 550 nm was measured. Using the absorbance obtained when water was used instead of the sample as a control (B), the inhibition rate (%) was determined using the following formula.

Inhibition rate (%) = 100X (B-A)/B (measurement
Results) Table 1 Test sample Inhibition rate (%) Extract of Example 1 Extract of Example 2 Non-adsorbed fraction of Example 3 20% of Example 3 Ethanol eluted fraction of Example 3 30% Ethanol Elution Fraction Example 5 Toothpaste: (Composition) Calcium diphosphate Glycerin Carrageenan Sodium lauryl sulfate Sodium saccharin Butyl paraoxybenzoate Light brown extract Non-adsorbed fraction 1 Flavored water (parts by weight) 0.9 1. 2 0.09 0.0 0 5 0.05 Total amount remaining 10
08 HP-20 non-adsorbed fraction obtained in Example 3.

Example 6 Mouth rinse liquid: (composition) (parts by weight) sodium lauryl sulfate glycerin Sorbitol Ethyl alcohol sweet tea extract statue 1-Menthol Incense fee saccharin sodium water Total amount 8 Extract obtained in Example 2.

Example 7 Lozenge: (If completed) gum arabic glucose Sweet tea extract storage Potassium phosphate 0.8 0.05 0.05 0.04 0.1 Remaining amount (parts by weight) 0.02 0.2 potassium phosphate Milk Sugar Incense fee Magnesium stearate Total amount Extract obtained in Example 1.

Fruit 1M Example 8 Gum: (Composition) Gum base Calcium carbonate Stevioside Light brown 30% ethanol, Fraction★ Lactose flavoring 0.1 0.1 Remaining amount (parts by weight) 0.1 0.01 76.89 Total amount 10
Of the light brown extract adsorption fraction obtained in Example 3, 30%
Ethanol elution fraction.

Example 9 juice: (string) Frozen concentrated Satsuma mandarin juice fructose glucose liquid sugar citric acid L-ascorbic acid Sweet tea 20% ethanol fraction Incense fee Color element water (parts by weight) 0.2 0.02 0.001 0.2 0.1 ” Quantity (composition) powder sorbitol Incense fee Sweet tea extract sorbitol seeds (parts by weight) 997 no 1 0.0 1 0.05 Total amount Extract obtained in Example 1.

(Left below) Total amount 10
0 Of the light tea extract adsorption fraction obtained in Example 3, 20%
Ethanol elution fraction.

Example 10 Sweetfish: Example 11 Bean paste: Example 12 Castella cake: Red raw red bean granulated sugar starch syrup light tea extract 8 Water 41.7 26.7 7.8 0.1 23.8 23.8 41.6 26. 7 7.8 Total amount 100.0 100
．． 01 Example 1. Either of the light tea extracts obtained in 2 or 3 may be used (the same applies to the following examples as well).

However, when using the 20% ethanol elution fraction obtained in Example 3, it may be around 1.15 to 172 as shown in the table).

cake flour whole eggs caster sugar water candy Light tea extract water Total amount Example 13 Is it water? 39.7 32.1 6.7 4.1 100.0 yeah: 17.24 39.7 32.1 6.7 0.16 4.1 100.0 Red bean paste powdered agar salt caster sugar Light tea extract water all amount 24.8 0.3 0.1 24.9 49.8 100.0 24.8 0.3 0.1 24.9 0.1 49.8 100.0 Example 14 Dorayaki skin: Example 16 Hatter cake: cake flour caster sugar whole eggs baking powder Light tea extract Total amount 33.1 33.1 33.1 0.7 100.0 33.0 33.1 33.0 O07 0,2 100,0 cake flour strong flour whole eggs margarine caster sugar Light tea extract all amount 20.0 5.0 25.0 25, ○ 25.0 100.0 20.0 5.0 24.9 25, O 25,0 0,1 100,0 Example 15 sponge cake: Example 17 Bavarois cake flour caster sugar whole eggs Light tea extract all amount 30.3 3”9,4 30.3 100.0 30.3 39.2 30.3 0.2 100.0 milk Fresh cream egg yolk Seratin caster sugar Light tea extract water 48.5 16.2 6.5 2.9 12.9 12.9 48.44 16.2 6.5 2.9 12.9 o, 06 12.9 all amount 100.0 100.0 Example 18 custard cream: fruit Example 20 Custard pudding: milk whole eggs cornstarch caster sugar Light tea extract all amount 66.2 13.2 4.0 16.5 100.0 66.12 13.2 4.0 16.5 0.08 100.0 milk whole eggs caster sugar water Light tea extract Total amount 47.6 31.9 17.1 3.4 1 00.0 47.51 31.9 17.1 3.4 0.09 100.0 Example 19 Buttercream: Example 21 Kutsky: milk margarine egg yolk Granulated sugar Light tea extract all amount 21.7 4”3.6 13.0 21.7 100.0 21.7 43.5 13.0 21.7 0.1 100.0 thin knife whole eggs margarine caster sugar baking powder water Light tea extract Total amount 31.9 Engineering 6.0 19.2 25.5 0.2 7.2 100.0 31.77 16.0 19.2 25.5 0.2 7.2 0.13 100.0 Example 22 sweet bread Example 23 steamed butter strong flour thin knife caster sugar isomerized sugar whole eggs shortening Dry yeast Food Salt water Light tea extract all amount 100.0 100.0 thin knife whole eggs Granulated sugar butter saladan white baking powder Light tea extract Total amount Example 24 jam: 26.2 32.8 26.2 6.6 5.9 2.3 100.0 26.2 32.67 26.2 6.6 5.9 2.3 0.13 100.0 Ichiko Granulated sugar citric acid pectin fragrance Light tea extract 54.8 42.0 0.6 2.4 0.1 54.69 42.0 0.6 2.4 0.1 0.21 all amount 100.0 100.0 Example 25 Example 26 Lactic acid bacteria drink: Carbonated drink: Milk solids content 21% fermented milk fructose wine liquid sugar pectin citric acid fragrance water Light tea extract Total amount 14.76 13.3 1 0.5 0.08 0.15 71.20 100.0 14.76 13.3 1 0.5 0.08 0.15 71.14 0.06 100.0 Granulated sugar concentrated lemon juice L-ascorbic acid citric acid sodium citrate coloring agent fragrance Carbonated water Light tea extract Total amount 100.0 100.0 husband Example 27 Coffee drinks: Example 28 Coffee celery: Granulated sugar skimmed milk powder caramel coffee extract fragrance polyglycerin fatty acid ester Food Salt water sweet tea extract all amount 8.0 5.0 0.2 2.0 0.1 0.05 0, o5 84.6 100.0 8.0 5.0 0.2 2.0 0.1 0.05 0.05 84.56 0.04 100.0 Granulated sugar Seven Latin coffee extract water Light tea extract all amount 15.0 1.0 5.0 79.0 15.0 1.0 5.0 78.93 0.07 100.0 100.0 Example 29 caramel: Granulated sugar water candy powder milk hydrogenated oil Food Salt fragrance water Light tea extract 32.0 20.0 4σ, 0 4.0 0.6 0.02 3.38 all amount 100.0 100.0 Example 30 ice cream Li Room: Example 32 juice: Fresh cream (45% fat) skimmed milk powder Granulated sugar sweetened egg yolk vanilla essence water Light tea extract Total amount Example blood Niyi hmm 33.8 11.0 14.8 0.3 0.1 40.0 100.0 Ga Mu: 33.8 11.0 14.8 0.3 0.1 39°93 0.07 100.0 frozen concentrated fruit juice sugar citric acid L-ascorbic acid fragrance color element water Light tea extract Total amount Example 33 candy 5.0 11.0 O02 0,02 0,2 0,1 83,48 100,0 100,0 gum base sugar fragrance citric acid water Light tea extract all amount 20", 0 76.0 1.0 1.0 2.0 100.0 20.0 75.62 1.0 1.0 2.0 0.38 100.0 water candy fragrance water Light tea extract all amount 50.0 1.0 2.0 100.0 49.76 1.0 2.0 0.24 100.0 Example 34 chocolate: all amount 100.0 100.0 Below Up

Claims (7)

[Claims] (1) Effective composition of aqueous solvent extract from sweet tea Anti-caries agent. (2) Synthetic absorption of aqueous solvent extract from sweet tea Obtained by adsorption treatment using adhesive Synthetic adsorbent non-adsorbed fraction as active ingredient Contains anti-caries agents. (3) Synthetic absorption of aqueous solvent extract from sweet tea Synthetic adsorption by adsorption treatment using adhesive adsorbed onto a solvent, and then adsorbed with a solvent. The synthetic adsorbent adsorption fraction obtained by desorption is Anti-caries agent containing as an active ingredient. (4) The anti-caries agent according to any one of claims 1 to 3, wherein the content of the active ingredient is 0.00001 to 0.5%. (5) Claim 1, wherein the dosage form is an oral hygiene agent. The defense mentioned in any of paragraphs 2 to 4 caries agent. (6) Any of claims 1 to 4 Food containing the anti-caries agent described in the above section. (7) After extracting sweet tea with an aqueous solvent, the extract is Subjected to adsorption treatment using a synthetic adsorbent, by the unadsorbed fraction and/or solvent. Obtain the desorbed synthetic adsorbent adsorption fraction. glucosyltrans which is characterized by Anti-cariogenic substance with ferase inhibitory activity Quality manufacturing method.