JP2887696B2 - Glucosyltransferase inhibitors and oral cleansing agents - 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 preventing caries by inhibiting the activity of a glucosyltransferase, an enzyme for synthesizing glucan that causes dental caries, and the glucosyltransferase. The present invention relates to an oral cleansing agent and a food and drink which are provided with a caries preventive action by adding an inhibitor.

[0002]

2. Description of the Related Art It is known that the occurrence of dental caries involves enzymes derived from microorganisms in the oral cavity, particularly enzymes and glucosyltransferases produced by Streptococcus mutans. In other words, glucosyltransferase produces water-insoluble and strongly adherent glucan from sucrose, which adheres to the tooth surface together with oral microorganisms to form plaque, and the sugar in food is metabolized by plaque microorganisms. Organic acid, which in turn erodes the enamel on the tooth surface and causes caries.

[0003] Therefore, in addition to removing plaque adhering to the tooth surface by using a toothpaste, etc., it suppresses the production of glucan by inactivating glucosyltransferase in the oral cavity, thereby preventing the formation of plaque. Is an effective means for preventing dental caries. From such a viewpoint, in recent years, oral cleansing agents such as toothpastes and foods and drinks containing a substance having a glucosyltransferase inhibitory action have been provided. As substances having a glucosyltransferase inhibitory action, conventionally, 1-desoxyxinojirimycin, N-methyl-desoxyxinojirimycin, crude drug tannins, ellagic acid, green tea polyphenol and the like are known.

[0004]

DISCLOSURE OF THE INVENTION The present invention relates to a novel glucosyltransferase inhibitor which is safe and easy to use from natural products, and an oral cleansing agent provided with a caries-preventing action using the same. It is intended to provide food and drink.

[0005]

SUMMARY OF THE INVENTION A novel glucosyltransferase inhibitor which has been successfully provided by the present invention is to extract the above-ground parts of a plant belonging to the genus Rose with water, a hydrophilic organic solvent, butanol, or a mixture thereof. It consists of an extract obtained by processing (hereinafter referred to as rose extract). The present invention also provides a caries-preventing oral cleansing agent and a food or drink, which contain the glucosyltransferase inhibitor according to the present invention as an active ingredient. In this specification, the term "food and drink" means not only food and drink for humans but also feed or feed for livestock and pet animals that can cause tooth decay.

Although it has not been confirmed yet what components in the rose extract are involved in the glucosyltransferase inhibitory action, the rose extract obtained by subjecting the rose extract to a fractionation treatment capable of concentrating the glucosyltransferase inhibitory component Can also be used as the glucosyltransferase inhibitor of the present invention.

[0007] Rose extracts having a glucosyltransferase inhibitory activity include plants belonging to the genus Rosa, such as Maikai, Hamanashi, Hamanasu, Damask rose, Neubara, Mokkobara, Koshinbara, Naniwaiba, Purple Rose and the like and roses thereof. By extracting the above-ground parts of hybrids or horticultural varieties from cultivars. Portions that are rich in active ingredients and are preferable as extraction raw materials are flowers, petals and buds, but leaves, stems and the like also contain active ingredients and can be used as extraction raw materials. In order to extract a component having a glucosyltransferase inhibitory action from the above-mentioned raw materials, the extract is dried and pulverized so as to be easily extracted, or a commercially available crude drug in the form of water or water at room temperature or in a heated state. What is necessary is just to immerse in a hydrophilic organic solvent. Examples of usable organic solvents include methanol, ethanol, acetone, and butanol, and these may be used as a mixture with water.

After the treatment, the extract is obtained by removing the solid matter by filtration or centrifugation, concentrated, and further dried if necessary. The rose extract used as the glucosyltransferase inhibitor of the present invention may be in any of the above-mentioned extract, its concentrated solution and dry powder. Since rose extract has some bitterness and is also hygroscopic, mixing excipients such as cyclodextrin, dextrin, and lactose facilitates the formulation operation and the storage and use of the formulation.

[0009] Since rose extract has an excellent glucosyltransferase inhibitory activity, it is effective in preventing dental caries when incorporated in a small amount in an oral cleansing agent such as toothpaste, mouthwash, and mouthwash. When added to foods and drinks containing a sugar sweetener, for example, soft drinks, confectionery bread, candy, chewing gum, gummy, jelly, etc., foods and drinks that are less likely to cause caries due to the sweetener can be obtained.

[0010]

EXAMPLES In the following examples, "parts" means parts by weight. Example 1 1000 ml of 50% ethanol was added to 100 g of crushed flower buds (dry matter) of a maikai, and the mixture was allowed to stand at 40 ° C. for 24 hours to extract soluble components. The obtained extract was concentrated to dryness under reduced pressure to obtain 25 g of a dried product. The same raw material was subjected to the same extraction treatment by changing the extraction solvent (including an example using a hydrophobic organic solvent for comparison).

Example 2 An extraction was performed in the same manner as in Example 1 by using crushed purple rose flower buds to obtain an extract.

Example 3 1000 ml of 50% ethanol was added to 100 g of dried pear flower (dry matter), and the mixture was heated under reflux for 1 hour to extract soluble components. The obtained extract was concentrated to dryness under reduced pressure to obtain 23 g of a dried product. The same raw material was subjected to the same extraction treatment by changing the extraction solvent (including an example using a hydrophobic organic solvent for comparison).

The glucosyltransferase inhibitory activity of the dried product obtained in Examples 1 to 3, ie, the rose extract, was measured by the following method. Measuring method of glucosyltransferase inhibitory activity: 2% sucrose solution (using 50 mM phosphate buffer at pH 6.5) 1.0 ml 1% sodium azide 0.1 ml crude glucosyltransferase solution 50 μl rose extract aqueous solution 50 μl (rose extract The enzyme reaction solution (rose extract content: 3.13 ppm) obtained by mixing each of the above solutions was placed in a test tube, and the test tube was placed in a tube. Incubate at 37 ° C for 20 hours. 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. Thereafter, 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. As a control, the same measurement is performed when water is added instead of the rose extract aqueous solution. 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 when rose extract is added; before enzyme reaction starts A ₁ : rose absorbance when the extract was added; glucan dispersion a _2: control; enzymatic reaction before the start a _3: control; shown in Table 1 to Table 3 glucan dispersion measurements.

Table 1 Glucosyltransferase activity inhibition rate of the rose extract of Example 1Extraction solvent Inhibition rate (%)  50% ethanol 84 water 65 ethanol 79 methanol 69 acetone 68 70% acetone 77 n-butanol 64 ethyl acetate (comparative example) 28 hexane (comparative example) 21 methylene chloride (comparative example) 26

TABLE 2 Glucosyltransferase activity inhibition rate of rose extract of Example 2Extraction solvent Inhibition rate (%)  50% ethanol 79 water 70 ethanol 67 methanol 68 acetone 70 70% acetone 75 n-butanol 50 ethyl acetate (comparative example) 19 hexane (comparative example) 12 methylene chloride (comparative example) 27

Table 3 Inhibition rate of glucosyltransferase activity of the rose extract of Example 3Extraction solvent Inhibition rate (%)  50% ethanol 72 water 71 ethanol 77 methanol 63 acetone 70 70% acetone 77 n-butanol 55 ethyl acetate (comparative example) 22 hexane (comparative example) 21 methylene chloride (comparative example) 17

EXAMPLE 4 Pulverized Maikai and Purple Lo
And 100 g of stems and leaves of raw grass
Thus, an extract was obtained by the following method. Extraction solvent: 1000 ml of 50% ethanol Extraction conditions: 40 ° C., 24 hours Treatment of the extract: the same as in Example 1 The obtained rose extract was treated with glucosyl as described above.
Table 4 shows the results of measuring the transferase inhibition rate.
You. Table 4Extracted material Inhibition rate (%)  Maikai leaf 57 Maikai stem 41 Purple rose leaf 61 Purple rose stem 39 Tsukushibara leaf 63 Tsukushibara stem 50

Example 5 Using 10 to 12 3-week-old ICR mice (♂) in each group,
After feeding the following feed for 30 days, the mandible was fuchsin-dissolved.
The solution was stained, and the state of the occurrence and progress of caries was determined. What
In addition, the rose extract used for the group D feed was obtained in Example 1.
It was obtained using 50% ethanol as the extraction solvent.
You. Group A: Normal feed (Charles River Japan Co., Ltd .; CRF-
1) Group B: Caries-inducing feed basic blended feed (0% sucrose concentration) Group C: Caries-inducing feed (30% sucrose concentration) Group D: Caries-inducing diet (1000% sucrose concentration) with rose extract
The results are shown in Table 4. The criteria for determining caries are as follows:
It is. Level 3: Extensive dentin with visible collapse of the cusps
Level 2: Dentin caries Level 1: Enamel caries Level 0: No caries recognized Table 4Test group Number of solids Number of test teeth Level 0 Level 1 Level 2 Level 3  A 11 22 14 800 B 10 20 16 4000 C 12 24 5 9 10 0 D 10 20 17 30 00

Example 6 The following raw materials were mixed by a conventional method for producing candy, and the mixture was further boiled and molded to produce a candy for preventing caries. Sucrose 70 parts Mizuame 30 parts Citric acid 1 part Fragrance 0.1 part Maikai extract 0.01 part Water 20 parts (Note) Maikai extract: 50% ethanol extract of Example 1

Example 7 The following raw materials were treated by a conventional method for producing chewing gum to produce an anti-carious chewing gum. Chewing gum base 20 parts Sucrose 55 parts Water syrup 20 parts Softener 4 parts Fragrance 0.8 parts Maikai extract 0.05 parts (Note) Maikai extract: 50% ethanol extract of Example 1

Example 8 The following ingredients were treated in the usual manner for chocolate production,
Anti-caries chocolate was produced. Chocolate 45 parts sucrose 15 parts cocoa butter 20 parts whole milk powder 25 parts purple rose extract 0.1 part (note) purple rose extract: 70% acetone extract of Example 2

Example 9 Dicalcium phosphate 45 parts Carboxymethylcellulose sodium 1 part Glycerin 20 parts Sodium lauryl sulfate 2 parts Flavor 1 part Glycyrrhizin 0.1 part Hamanashi flower extract 0.1 part Water 30 parts (Note) Hamanashi no Flower extract: 50% ethanol extract of Example 3 The above ingredients were mixed to produce a caries preventive mouthwash.

Example 10 Flour 30 parts Cornflower 15 parts Soy flour 15 parts Sucrose 5 parts Tallow 5 parts Salt 1 part Calcium phosphate 2 parts Calcium sorbate 0.5 part Fragrance 0.5 parts Maikai flower bud extract 0.1 Part (Note) Extract of Maikai flower buds: 50% ethanol 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 further mixed, and then pelletized with an extruder. To produce dog food.

[0024]

According to the present invention, natural products are used as raw materials,
A novel glucosyltransferase inhibitor which is safe and easy to use is provided. Since there is no concern if this inhibitor is taken into the body, it is possible to provide not only an oral cleansing agent imparted with a caries-preventing action but also a caries-preventive food or drink by incorporating it.

──────────────────────────────────────────────────続 き Continuation of front page (72) Inventor Akemi Tateno 14703-10 Mukotocho, Onomichi-shi, Hiroshima Maruzen Kasei Co., Ltd. (56) References JP-A-4-202107 (JP, A) JP-A-4-178230 (JP, A) JP-A-4-99712 (JP, A) JP-A-4-66534 (JP, A) JP-A-3-176418 (JP, A) JP-A-3-101687 (JP, A) Kaihei 1-301614 (JP, A) (58) Field surveyed (Int. Cl. ⁶ , DB name) A61K 7/26 A23G 3/00 A23L 1/03

Claims (3)

(57) [Claims] 1. A glucosyltransferase inhibitor comprising an extract obtained by subjecting an aerial part of a plant belonging to the genus Ros to an extraction treatment with water, a hydrophilic organic solvent, butanol, or a mixture thereof. 2. A caries-preventing oral cleansing agent comprising the glucosyltransferase inhibitor according to claim 1 as an active ingredient. 3. A food or drink, to which the glucosyltransferase inhibitor according to claim 1 is added.