JP3281716B2 - Glucosyltransferase inhibitors, oral preparations and foods and drinks - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a glucosyltransferase inhibitor having an action of inhibiting the activity of glucosyltransferase, an enzyme for synthesizing glucan that causes dental caries, to prevent the occurrence of cavities, and the inhibition of the glucosyltransferase The present invention relates to an oral agent and a food and drink which are provided with a caries preventive action by adding an agent. In this specification, foods and drinks are meant to include not only foods and drinks for humans but also feeds or feeds for livestock and pets that may cause dental caries.

[0002]

2. Description of the Related Art The development of dental caries involves an oral microorganism, particularly an enzyme glucosyltransferase produced by Streptococcus mutans. In other words, sucrose remaining in the oral cavity of foods and drinks is changed into water-insoluble and strongly adherent glucan by the action of glucosyltransferase, and adheres to the tooth surface together with the oral microorganisms to form plaque. . Microbes in the dental plaque metabolize sugars in the food to form an acid, and this acid demineralizes and erodes tooth enamel.

[0003] Therefore, in order to prevent dental caries, plaque adhering to the tooth surface is not only removed by using toothpaste or the like, but also by inhibiting the growth of Streptococcus mutans and the action of glucosyltransferase in the oral cavity. It is most effective to suppress the formation of glucan and thus prevent the formation of plaque.

[0004] From such a viewpoint, in recent years, oral preparations such as toothpastes and foods and drinks containing substances having a glucosyltransferase inhibitory action have been provided.
Conventionally known glucosyltransferase activity inhibitors useful for such applications include mutastain, crude drug tannins, ellagic acid, green tea polyphenol, oolong tea extract and the like.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel glucosyltransferase inhibitor made from a highly safe natural product, an oral agent having a caries-preventing action by adding the same, and a food and drink. To provide things.

[0006]

The glucosyltransferase inhibitor successfully provided by the present invention is a plant belonging to the legume family, tamarind (Tamarindus indica L.).
The substance extracted from the tamarind husk, which is the part of the seed skin, is used as an active ingredient. The present invention also provides an oral agent and a food or drink characterized by containing the glucosyltransferase inhibitor.

[0007] Tamarind husk is a part of husk (seed coat) that is separated and discarded when producing tamarind seed gum, which is used as a thickener in food processing, from the endosperm part of tamarind seeds. It was not previously known that a substance having an inhibitory action was contained.

The glucosyltransferase activity inhibitor contained in tamarind seeds is water or other aqueous solvents;
C 1-4 lower aliphatic alcohols such as methanol, ethanol, isopropanol and butanol; polyhydric alcohols such as 1,3-butylene glycol, propylene glycol and glycerin; lower aliphatic ketones such as acetone and methyl ethyl ketone; When the tamarind husk is immersed in the mixture at room temperature or in a heated state, it is efficiently extracted. aliphatic hydrocarbons such as n-hexane,
Halogenated hydrocarbons such as methylene chloride and chloroform;
Extraction is possible using a lower aliphatic ester such as ethyl acetate, but the extraction efficiency is not so good, and the obtained extract has a slightly weaker enzyme activity inhibiting effect.

A solid substance is separated from the extract by filtration or centrifugation, the solvent is distilled off, and if necessary, further drying is performed. The extract concentrate and the dried product thereof exhibit a strong glucosyltransferase activity inhibitory activity, and can be used as such as the glucosyltransferase inhibitor of the present invention.

[0010] It has not been confirmed what component in the tamarind husk extract actually inhibits the action of glucosyltransferase. However, since the active ingredient is not removed by the degree of treatment with activated carbon, the extract contains activated carbon. Deodorization and decolorization may be performed. Further purification can be performed, if necessary, as long as it is effective for improving the enzyme inhibitory activity.

The tamarind husk extract not only inhibits the activity of glucosyltransferase, but also exhibits an antimicrobial activity against the aforementioned cariogenic bacterium, Streptococcus mutans, which is not very strong (minimum inhibitory concentration MIC: About 500-2000 μg / ml). Therefore, the tamarind husk extract is effective in preventing the production of glucan by both the above-mentioned enzyme activity inhibitory action and antibacterial action.

[0012] The glucosyltransferase inhibitor of the present invention includes, in addition to the extract obtained by extracting the glucosyltransferase activity inhibitor from tamarind husk as described above or its purified product, any auxiliary agent, shaping Water or an organic solvent for use as an agent or a solution can be appropriately contained. For example, the tamarind husk extract has some astringency, but when this astringency becomes an obstacle in using the glucosyltransferase inhibitor of the present invention in oral preparations and foods and drinks, cyclodextrin, dextrin, Lactose, sugar alcohol (eg, sorbitol, maltitol, xylitol, erythritol, etc.) can be mixed to mask the astringency.

The glucosyltransferase inhibitor of the present invention can be added to oral preparations and its glucosyltransferase inhibitory action can be utilized to prevent glucan production in the oral cavity. Examples of suitable oral preparations to be added include various toothpastes, mouthwashes, troches, chewing gums, oral pasta, gingival massage creams, gargles, mouth fresheners and the like.

The addition of the glucosyltransferase inhibitor of the present invention does not limit the other components of the oral preparation or the method for producing the oral preparation. For example, calcium hydrogen phosphate, calcium carbonate, insoluble sodium metaphosphate,
Abrasives such as aluminosilicate, silicic anhydride, resin;
Surfactants such as long-chain sodium alkyl sulfate, sodium lauryl sulfoacetate, lauryl diethanolamide, sucrose fatty acid ester; CMC, hydroxyethyl cellulose, alginate, carrageenan, gum arabic,
Binders such as polyvinyl alcohol; thickeners such as polyethylene glycol, sorbitol, glycerin, propylene glycol; sweeteners such as saccharin, steviosides, glycyrrhizic acid, thaumatin; preservatives such as dehydroacetic acid, sodium dehydroacetate; menthol; Fragrances such as carvone, eugenol, anethole, peppermint oil, spearmint oil, peppermint oil, eucalyptus oil, ginger oil, anise oil; various pigments and other raw materials commonly used in the manufacture of oral agents, depending on the type and use of the product Arbitrarily select,
It can be manufactured by an ordinary method.

When an oral preparation is prepared by adding the glucosyltransferase inhibitor of the present invention, another glucosyltransferase inhibitor may be used in combination.
An antibacterial agent effective against Streptococcus mutans may be used in combination. Further, by adding an optional anti-inflammatory agent, antibacterial agent, deodorant and the like, a more excellent oral agent can be provided. Examples of anti-inflammatory agents that can be added include Asenyaku, Licorice, Uwaurushi, Ogon, Kouki, Psycho, Hawthorn, Perilla, Paeonia, Souhakuhi, Kyonin, Tissou, Clove, Tonin, Nikuzuku, Buttonpi, Azulene, Allantoin, Ursolic acid, Examples include oleanolic acid, glycyrrhizic acid, glycyrrhetinic acid, tocopherol, tranexamic acid, and the like.Examples of antibacterial agents that can be added include gobishi, saishin, sansho, ginger, dill, thyme, rosemary, and ascorbic acid. , Mustatin, humic acid, linoleic acid, linolenic acid and the like. Further, examples of the deodorant that can be used in combination include: amateur, fennel, vulgaris, ellagic acid, calyx, pepper, mace, sage, perilla, ginkgo, oyster leaves, green tea, oolong tea, pepper, rosin, oyster astringent, actizol, Chlorophylline derivatives, chlorhexidine, Maillard reactants and the like can be mentioned.

The tamarind husk extract which is an active ingredient of the glucosyltransferase inhibitor of the present invention, when dissolved in water, gives a homogeneous solution easily at a practical concentration. There is no point.

Since the effectiveness of the glucosyltransferase inhibitor of the present invention varies depending on the content of the active ingredient, it is difficult to uniformly show a suitable amount of the glucosyltransferase inhibitor of the present invention to an oral preparation.
Describing the case where the standard tamarind husk extract is directly blended into toothpaste or the like, about 0.001 to
An appropriate amount is 5.0% by weight, and a particularly preferable compounding ratio is about 0.5%.
005 to 1.0% by weight.

The glucosyltransferase inhibitor of the present invention can also be added to foods and drinks containing sucrose to prevent the formation of glucan in the oral cavity and thus prevent caries. Examples of suitable additives include soft drinks, confectionery, bread, candy, chewing gum, gummy, jelly, chocolate, pet food, and the like.

[0019]

The present invention will be described below with reference to examples. In each of Examples 2 to 7, “parts” means parts by weight. Example 1 100 g of crushed tamarind husk was added to 100 g of extraction solvent.
0 ml was added, and the mixture was allowed to stand at 40 ° C. for 24 hours to extract soluble components. The obtained extract is concentrated to dryness under reduced pressure to obtain a solid extract. The same extraction procedure was performed using the same tamarind husk and only the extraction solvent was varied, and the resulting extract was examined for the inhibition of glucosyltransferase activity by the following method.

Glucosyltransferase activity inhibition rate measurement method: 2% sucrose solution (using 50 mM potassium phosphate buffer at pH 6.5) 1.0 ml 1% sodium azide 0.1 ml crude glucosyltransferase solution 50 μl aqueous solution of tamarind husk extract (Concentration: about 2 to 0.0005 mg / ml) 50 µl 50 mM potassium phosphate buffer, pH 6.5 0.1 ml The enzyme reaction solution obtained by mixing the above solutions is put into a test tube, and the test tube is tilted at 30 °. At 37 ° C for 20 minutes.
Let stand for a while. Thereafter, the test tube is slowly tilted to remove the reaction supernatant, and the glucan produced by the enzyme reaction and attached to the test tube is washed three times with water. Then, 2 ml of water is added, and the glucan is dispersed in water by sonication. The absorbance at a wavelength of 550 nm is measured for the glucan dispersion and the enzyme reaction solution immediately after preparation. Separately, as a control, the same operation is performed when water is added instead of the aqueous solution of the tamarind husk extract. From the measurement results, the inhibition rate of glucosyltransferase activity is calculated by the following formula.

Inhibition rate (%) = [1- (A ₁ -A ₀ ) / (A
_3- A ₂ )] × 100 where A ₀ : absorbance before the start of the enzyme reaction A ₁ : absorbance of the glucan dispersion A ₂ : control (before the start of the enzyme reaction) A ₃ : control (glucan dispersion)

[0022] From the results of the test, the (strong enzyme inhibitory activity as an IC ₅₀ value is small) tamarind husk extract concentration was determined as IC ₅₀ value when a 50% inhibition. Table 1 shows the results.

[0023]

Table 1 Extraction solvent Extraction yield (%) IC ₅₀ value (μg / ml) 50% ethanol 30.0 0.085 80% ethanol 29.0 0.052 water 13.2 0.16 ethanol 32.6 0.065 Methanol 34.1 0.060 Acetone 22.5 0.055 50% Acetone 30.0 0.045 70% Acetone 27.3 0.042 Ethyl acetate 9.3 0.43 n-Hexane 0.90 .63 methylene chloride 0.4.48 n-butanol 15.7 0.103 50% 1,3-butylene glycol 21.3 0.079

Example 2 The following raw materials were mixed by a conventional method for producing candy, and the mixture was further boiled down to form a candy having caries prevention properties. 70 parts sucrose 30 parts starch syrup 30 parts citric acid 1 part Fragrance 0.1 part Tamarind husk extract 0.01 part (Tamarind husk extract: hexane extract of Example 1)

Example 3 The following raw materials were treated by a conventional method for producing chewing gum to produce a caries-preventing chewing gum. Chewing gum base 20 parts sucrose 55 parts starch syrup 20 parts softener 4 parts incense (mint oil) 0.8 parts tamarind husk extract 0.05 parts (tamarind husk extract: acetone extract of Example 1)

Example 4 The following ingredients were treated in the usual manner for the production of chocolate,
Anti-caries chocolate was produced. Chocolate 45 parts sucrose 15 parts cocoa butter 20 parts whole milk powder 25 parts tamarind husk extract 0.1 part (tamarind husk extract: 50% ethanol extract of Example 1)

Example 5 Dicalcium phosphate 45 parts CMC sodium salt 1 part Glycerin 20 parts Sodium lauryl sulfate 2 parts l-Menthol 1 part Glycyrrhizin 0.1 part Tamarind husk extract 0.1 part Water 30 parts (Tamarind husk extraction Product: Ethanol extract of Example 1) The above-mentioned raw materials were mixed to produce a caries-preventive toothpaste.

Example 6 Ethanol 20 parts Glycyrrhizin 0.2 parts Glycerin 5 parts Chlorhexidine 0.005 parts l-Carbon 1 part Tamarind husk extract 0.5 part Water 70 parts (Tamarind husk extract: water extraction of Example 1) Product) The above-mentioned raw materials were mixed to produce a caries preventive mouthwash.

Example 7 Flour 30 parts Corn flour 15 parts Soybean flour 15 parts Meat meal 20 parts Sucrose 5 parts Tallow 5 parts Salt 1 part Calcium phosphate 2 parts Potassium sorbate 0.5 part Fragrance 0.5 parts Propylene glycol 7 parts 0.1 part of tamarind husk extract (Tamarind husk extract: 50% acetone extract of Example 1) The above raw materials were mixed, and 40 parts of water was added to 100 parts of the obtained mixture, and the mixture was heated with an extruder. While being molded into pellets, dog food was manufactured.

[0030]

EFFECT OF THE INVENTION The glucosyltransferase inhibitor of the present invention, which contains a glucosyltransferase activity inhibitory substance extracted from the husk portion of tamarind, which has been used as a raw material for food production, as an active ingredient, is highly safe and is taken into the body. Don't worry if you do. In addition, since the tamarind husk extract itself shows a strong enzyme activity inhibitory action, the amount of addition required for imparting a caries preventive action by adding it to oral preparations or foods and drinks is extremely small. Therefore, the use of the glucosyltransferase inhibitor of the present invention makes it easier to produce a caries-preventing oral preparation and food / drink than ever before, which can contribute to reducing dental caries.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ identification code FI A61P 43/00 111 A61P 43/00 111 C12N 9/99 C12N 9/99 (58) Fields surveyed (Int.Cl. ⁷ , DB Name) A61K 35/78 A23G 3/30 A23L 1/30 A61K 7/26 C12N 9/99 BIOSIS (DIALOG) CA (STN)

Claims (4)

(57) [Claims] 1. A glucosyltransferase inhibitor comprising, as an active ingredient, a glucosyltransferase activity inhibitor extracted from tamarind husk. 2. An oral preparation comprising the glucosyltransferase inhibitor according to claim 1. 3. A food or drink comprising the glucosyltransferase inhibitor according to claim 1. 4. A glucosyltransferase activity inhibitor, wherein tamarind husk is subjected to extraction treatment with water, monohydric or polyhydric lower aliphatic alcohol, lower aliphatic ketone, halogenated hydrocarbon, aliphatic hydrocarbon, or a mixture thereof. The glucosyltransferase inhibitor according to claim 1, which is an extract obtained by the above method.