JPH0225413A - Preventive for dental caries - Google Patents

Abstract

PURPOSE:To obtain a preventive for dental caries having high safety by compounding crude catechin extracted from tea leaves, fraction separated from crude catechin, fraction for crude catechin ester or pure catechin. CONSTITUTION:The objective agent is produced by compounding crude catechin extracted from tea leaves (e.g., green tea, oolong tea or black tea), fraction separated from crude catechin, fraction of crude catechin ester extract or pure catechin produced by purging those crude catechin, etc., as an active component, properly mixing the active component with conventional components such as abrasive, binder, wetting agent, forming agent, essential oil, flavor, sweetener, preservative and water and forming in the form of a drug by conventional method, It is used in the form applicable to oral cavity, e.g., dentifrice and mouthwash (e.g., toothpaste). It is effective in exterminating or suppressing the proliferation of Streptococcus mutans and remarkably preventing the generation of caries.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a dental caries preventive agent that can prevent dental caries by sterilizing and suppressing the proliferation of cariogenic bacteria in the oral cavity.

(Prior technology) Streptococcus minitans, which is considered to be a cariogenic bacterium, attaches to a thin film called becryl present on tooth enamel, and synthesizes water-insoluble glucan, a polysaccharide, from sucrose. Water-soluble glucan forms dental plaque with a bacterial flora that also includes Streptococcus mutans and other bacteria. Streptococcus mutans demineralizes the enamel surface by fermenting various sugars and producing acids. Additionally, plaque prevents acid from diffusing into saliva, further accelerating demineralization.

Therefore, in order to prevent dental caries, it is desirable to suppress the number of bacteria of Streptococcus mutans, which is a major cause, or to suppress or inhibit dental plaque formation.

Conventionally, caries prevention agents include plaque formation inhibitors, acid-resistant agents,
There are oral bacteria inhibitors, and chlorhexidine gluconate, chlorhexidine hydrochloride, etc. are used as oral bacteria inhibitors.

(Problem to be solved by the invention) In recent years, various additives have been added to chemically synthesized products, such as tocopherol, which is vitamin E, being frequently used as an antioxidant due to growing concerns among consumers about chemically synthesized products. There is a movement towards methods that use naturally derived compounds.

Therefore, it seems that a safer oral bacteria inhibitor is needed as a caries preventive agent that is likely to be ingested through the oral cavity.

(Means for Solving the Problems) Therefore, as a result of intensive research, the present inventors have determined that crude catechin or crude catechin free type fraction extracted from tea leaves, crude catechin ester type fraction, or pure catechin obtained by purifying them. It was discovered that this method is effective in preventing erosion, and the present invention was developed based on this finding.

Therefore, the present invention provides crude catechin, crude catechin free fraction,
The gist of the present invention is an anti-caries agent characterized by using a crude catechin enamel type fraction or pure catechin.

(Effect) According to the present invention, these catechins sterilize or suppress the growth of Streptococcus mutans.

(Example) The tea leaves from which crude catechin, crude catechin free fraction, crude catechin ester fraction, or pure catechin used in the present invention are extracted are not particularly limited as long as they are incorporated into tea. Instead, it is also possible to use tea such as unfermented green tea, semi-fermented tea oolong tea, fermented tea black black, or unprocessed tea leaves.

An example of a method for extracting and producing crude catechin, crude catechin free type fraction, and crude catechin ester type fraction is as follows. Further, to purify pure catechin, these extracted crude catechins are purified by high performance liquid chromatography.

Collect the juice obtained by extracting 1 kg of tea with 20 Il of boiling water for 30 minutes and squeezing it, inject it into a cylindrical column packed with 500 g of hydroxylpropylated dextran gel, and let it flow down immediately by gravity or suction. I let it happen. After the panoramic injection, the column was washed with 3J distilled water to remove caffeine, amino acids, sugars, etc. Next, a 15% hydrophilic organic solvent aqueous solution 31 was allowed to flow down to wash out terpenoids, brown pigments, and brown tannin oxides. Thereafter, a 30% hydrophilic organic solvent aqueous solution 3β was allowed to flow down to elute the A fraction containing andocyanide, L-epigallocatechin, and L-epicatechin from the gel column, and then a 60% hydrophilic organic solvent aqueous solution 31 Flowing down, L epigallocatechin gallate,
The B fraction containing L-epicatechin gallate was eluted from the gel column. These fractions were collected and spray-dried to produce tea catechin compounds.

The above fractionation state is as shown in Figure 1,
The figure shows a gel column chromatography column (Se) using tea juice.
phadexLII-20 column chromatography). In the figure, Δ is the above A fraction, B is the above B fraction,
The dotted line indicates methanol concentration.

Further, FIG. 2 shows a high performance liquid chromatogram of the A fraction, and FIG. 3 shows a high performance liquid chromatogram of the B fraction.

Fraction A contains 97.8% L-epigallocatechin (code 1
) and a small amount of L-epicatechin, and the B fraction contains 77.2% epigallocatechin gallate (code 2) and 14.6% epicatechin gallate (code 3). was included.

The yield of crude catechin compounds from the extract according to the above example was as shown in Table 1.

Table 1 Note that the conditions for juice extraction in the above are not particularly limited. Hydrophilic vinyl polymer gel or the like can be used instead of the hydroxylpropylated dextran gel. Strict substances can be sufficiently removed by sequentially washing the column with distilled water and a hydrophilic organic solvent, and the concentration of the latter is preferably about 5 to 15% for its washing effect. Extraction of A fraction is 20-30% above.
If the concentration of the hydrophilic solvent is too dilute, it will be difficult to carry out the fractionation (with most of the A fraction coming out within the above concentration range, and if it is too concentrated than the above range, the B fraction will be difficult to perform). The fractions will be mixed and the fractionation will be insufficient.The extraction of the B fraction is maximum at around 40 to 65%, and if it is too thin than this level, the fractionation with the A fraction will be insufficient. In a region that is too dark, there is almost no remaining amount of the B fraction, and the functionality of the fraction is lost.

The dental caries preventive agent according to the present invention can be adjusted in various ways and used in various products applied to the oral cavity, such as toothpastes and mouthwashes.

For example, in toothpaste, dibasic calcium phosphate, calcium carbonate, calcium birophosphate, insoluble sodium metaphosphate, amorphous silica, crystalline silica, aluminosilicate, aluminum oxide, abrasives such as resin, sodium carboxymethyl cellulose, hydroxyethyl cellulose, alginate, etc. , caranagen, gum arabic, binders such as polyvinyl alcohol, wetting agents such as polyethylene glyconopesorbitol, glycerin, propylene glycol, sodium lauryl sulfate, sodium dodecylbenzenesulfonate, hydrogenated coconut fatty acid monoglyceride sodium monosulfate, lauryl sulfoacetic acid. Foaming agents such as sodium, sodium N-lauryl sulfoacetate, N-amylglutamate, sucrose fatty acid ester,
In addition, essential oils such as peppermint and spearmint, fragrance materials such as l-menthol, carboeugenol, and anethole, sodium saccharin, stevioside,
It is produced according to a conventional method by mixing components such as sweeteners and preservatives such as neohesperidyl dihydrochalcone, clycyrrhizin, perillartin, and P-methquin cinnamic aldehyde with water. Also, in mouthwashes and other mouthwashes, ingredients are appropriately added depending on the properties of the product.

Examples of these are shown as Example 11 and Example 2.

Example 1 - Toothpaste formulation example Hydrogen phosphate 50.0 D-Sorbitol 10.0 Sodium lauryl sulfate 20 Sodium Cal, f, quinmethylcellulose 10 Sodium saccharin 01 (wt%) Flavor crude catechin ester type fractionated water Example 2 - Mouthwash formulation example Ethanol Sodium lauryl sulfate Sodium saccharin Flavor Crude catechin ester type Fraction Water balance 20, fl (wt%) 0.05 1.0 2.0 Balance (experimental example) (1) Experimental materials and test bacteria crude Catechin free type fraction and crude catechin ester type fraction were extracted and produced according to the above example.

Further, crude catechin was prepared by mixing these free type fractions and ester type fractions at a ratio of 1:1. The pure catechin used was one in which only (-)epigallocatechin gallate was purified from the ester fraction by high performance liquid chromatography.

As the test bacterium, Streptococcus mutans swingbrit, which belongs to serotype C, which is abundant in the human oral cavity, was used.

(2) Test of lowest concentration of catechin inhibiting growth of Streptococcus mutans The assay method used a liquid medium dilution method. First, the medium prepared for catechin (Y AST NlTR0GEN B to SB
l11108 MINDACIDS 4g, DEX
TRO3 [42.5g, ammonium sulfate 2.5g,
Polypeptone 31g! Potassium monophosphate 0.5g,
Adjust the pH of 1000- of water to 7.1 with sodium hydroxide)
Put 9/10 amount of the sample into a test tube, add a dilution series of the sample prepared separately, inoculate test bacteria drop by drop after culturing in Plain Heart Infusion Broth medium for 24 hours, and incubate these for 24 hours 37 days. Cultured at ℃. Growth inhibition was determined by measuring O, D, -1,0660 nm (D absorbance). The results are shown in Table 2.

Table 2 6) Assay of bactericidal activity of Streptococcus mutans by catechin Streptococcus mutans was inoculated into the above-mentioned medium prepared for catechin, cultured for 24 hours, and the initial number of bacteria was counted on a plain heart infusion agar medium. and add a dilution series of the sample to the culture solution.
After 3 minutes, the number of bacteria after addition was counted on the plate.

The results are shown in Table 3.

Table 3: Not proliferating ± Slightly proliferating+, Slightly proliferating ・Proliferating, Extremely proliferating (4) Toothbrushing against the lowest concentration of catechin that inhibits the growth of Streptococcus mutans Effects of major components Add a dilution series of the crude catechin ester type fraction to the medium prepared for catechin as described above in the same manner as in the minimum growth inhibition concentration assay, and then add a dilution series of the toothpaste ingredients to be assayed. The test bacteria were inoculated in the same manner as the minimum growth inhibition concentration test, and after culturing at 37°C for 24 hours, O, D, = 1.0.660.
The determination was made by measuring the absorbance at nm. The results are shown in Table 4.

From the results in Tables 2 and 3 of Table 4, it is clear that the crude catechin, crude catechin free fraction, crude catechin ester fraction, or pure catechin of the present invention effectively sterilizes or inhibits the growth of Streptococcus mutans. From the results in Table 4, it is found that ingredients such as toothpaste do not affect the oral bacteria suppressing effect of catechin.

The above-mentioned catechins can suppress the proliferation of Streptococcus minitans when added at a concentration of 0.01 to 0.02% by weight, and bring about a remarkable reduction effect when added at a concentration of about 0.5% or more. A nearly sterile state can be created by mixing the ingredients to a certain degree.

(Effects of the Invention) As described above, the caries preventive agent of the present invention sterilizes or inhibits the proliferation of Streptococcus mutans by using crude catechin, crude catechin free fraction, crude catechin ester fraction, or pure catechin. Good prevention of caries. Moreover, crude catechin, crude catechin free fraction, crude catechin ester fraction, or pure catechin are natural products and are made from tea leaves that have been drunk for a long time.
The caries preventive agent of the present invention has extremely high safety in use.

[Brief explanation of the drawing]

Figure 1 is a gel column chromatogram of tea juice, Figure 2
The figure shows the composition of the A fraction, and FIG. 3 shows the composition of the B fraction. Itoen Co., Ltd. Figure 2■ Figure 3

Claims (1)

[Claims] A caries prevention agent characterized by containing crude catechin or a crude catechin free fraction extracted from tea leaves, a crude catechin ester fraction, or pure catechin obtained by purifying them.