JPH0495020A - Enzyme inhibitor - Google Patents

Abstract

PURPOSE:To provide an enzyme inhibitor employed in a oral treatment for preventing the generation of dental caries and periodontitis by compounding the solvent extract of a specific plant. CONSTITUTION:An enzyme inhibitor containing one kind or more of the solvent extracts of plants such as Paeoniae Radix and Eriobotrya japonica Lindl (preferably extracts using ethyl alcohol and/or water). The solvent extract inhibits glucosyl transferase originated from Streptococcus mutans and effectively prevents the formation of bacterial plaques.

Description

[Detailed Description of the Invention] <Industrial Application Field> The present invention relates to an enzyme inhibitor and an oral composition, and more particularly to an oral composition containing a solvent extract of a plant selected from Peony, Biwayoum, J. chinensis, and Chiyu. Regarding.

<Prior Art> Approximately two-thirds of the dental plaque that adheres to the surface of teeth is composed of microorganisms, and has an adverse effect on the oral cavity through the various metabolic activities of these microorganisms. In other words, dental plaque can cause tooth decay (caries) and periodontal disease (alveolar pyorrhea, etc.), which are said to be two diseases in dentistry.
Not only is it the culprit for bad breath, it is also a breeding ground for anaerobic bacteria that cause bad breath. In the formation of this dental plaque, glycoproteins in saliva adhere to the tooth enamel surface and form a thin film (approximately 1 μm in thickness) called a pellicle, which causes Streptococcus mutans (
Streptococcus mutans adheres due to the interaction between the bacterial cell surface substance and the pellicle. It then attaches via a unique insoluble glucan from Streptococcus mutans. This insoluble glucan has properties like a kind of adhesive, and it adheres strongly to the tooth, containing many bacteria in addition to the weakly attached Streptococcus mutans, which does not have the above-mentioned insoluble glucan. Forms dirt. This insoluble glucan has a main chain of α(1→3) glycosidic bonds, a side chain of α(1→゛6) glycosidic bonds, and is formed using sucrose present in the oral cavity. At this time, glucosyltransferase (referred to as dextran synthase double-subordinate GTF) possessed by Streptococcus mutans is involved, and mutan or levan is produced as an insoluble dextran.

A purification method for GTF has finally been established in recent years, and the serotype d/g/
Streptococcus mutans of m is 3-4 species of GT
It has been confirmed that one of these enzymes is an enzyme that synthesizes insoluble glucan, and the rest are enzymes that synthesize water-soluble glucan.

Methods aimed at anti-GTF action and decomposition of insoluble glucan include oral compositions containing enzymes such as α(1→3) glucanase and mutastein to decompose sugars (insoluble glucans, etc.) attached to teeth; Along with the development of chewing gum, extracts extracted from plants (e.g. Genso nato) using organic solvents (JP-A-57-5861O, JP-A-61-26001) were developed.
No. 2) The anti-GTF effect of an extract extracted with water or lower alcohol from an Ayurvedic medicinal herb used in Sri Lanka (Japanese Patent Application Laid-Open No. 60-54312) has been studied.

As mentioned above, many studies have been conducted on the anti-GTF effects of crude drugs, but not all of them have been studied. Peonies are already known for their sedative, analgesic, and
It has been confirmed to have pharmacological effects such as antispasmodic, anti-inflammatory, blood pressure lowering, and vasodilation, and is used for antispasmodic, analgesic, relaxation, and astringent purposes. In addition, Biwayo is used as an antitussive, expectorant, hot water, etc., and Jiyouintin is used as a piling agent, anthelmintic, and has a choleretic effect, and is used to treat hepatitis,
It has been used to treat inflammatory conditions, hives, and swelling, as well as as an inflammatory diuretic and choleretic, and chuyu has been used for purposes such as astringency, hemostasis, hot water, analgesia, and anti-inflammation.

<Problems to be solved by the invention> However, research on the anti-GTF effects of crude drugs that have been used for various purposes is still insufficient. Therefore, as a result of further investigating the anti-GTF effect of Streptococcus mutans on a wide variety of herbal medicines,
It has been newly discovered that peony, chinensis, chinensis, and chinensis have anti-GTF activity. In other words, solvent extracts of plants such as Peony, Biwayo, Shuinchin, and Chiyu suppress plaque formation caused by Streptococcus mutans and are suitable for preventing tooth decay or periodontal disease; Invented a new enzyme inhibitor.

<Means for Solving the Problems> The present invention provides an enzyme inhibitor characterized by containing one or more solvent extracts of plants selected from Peony, Biwayo, J. chinensis, and Chiyu, and the above-mentioned enzyme inhibitor. It relates to an oral composition characterized by containing as an active ingredient.

The enzyme inhibitor according to the present invention includes toothpaste, toothpaste, water toothpaste,
Used as mouthwashes, troches, tablets, powders, liniments, mouthwashes, mouth fresheners, chewing gum, drinks, candies, etc.The active ingredients are selected from peony, loquat, shoinchin, and chuyu. It is made by blending one or more kinds of solvent extracts of plants.

Solvents used for extracting the plants include water, polar solvents such as alcohol acetone, ethyl ether, and ethyl acetate;
Non-polar solvents such as benzene, hexane and petroleum ether can be used, and mixed solvents such as hydrous alcohols can also be used. In particular, alcohols include lower alcohols having 1 to 5 carbon atoms, such as methyl alcohol, ethyl alcohol, propyl alcohol, isopropyl alcohol, butyl alcohol, and amyl alcohol.

Examples of hydrous alcohols include hydrous methyl alcohol and hydrous ethyl alcohol, which may be used in combination of one or two types.
It can also be used as a mixture of more than one species.

Among these solvents, ethyl alcohol, water, or a mixed solvent thereof is preferable because it can simplify the conventional process of removing solvents that do not have a positive effect on the human body by operations such as vacuum concentration. When such a solvent is used, it is possible to create a composition that is highly safe against accidental ingestion.

The solvent extract used as an active ingredient in the present invention is obtained by extracting one or more of the above-mentioned plants using one or more of the above-mentioned solvents and removing the residue from the extract. obtain.

As the extraction method, not only various conventional methods such as steam distillation can be used, but also simply immersing the plant in a solvent for a long time. Further, the plant may be used for extraction as it is, but the extraction efficiency is better if it is used after being crushed or powdered.

The anti-GTF effect of each plant mentioned above will be explained in detail based on the following test examples. Furthermore, as a control plant, Gensou (Genso), whose solvent extract has been known to have an anti-GTF effect, was used.

Test Example 1: Effect of solvent extracts of plants A solvent extract of plants was produced and GTF for testing was prepared by the method described below, and the anti-GTF effect of each solvent extract was measured by an insoluble glucan adhesion inhibition test. did.

The amount of insoluble glucan was expressed in terms of glucose, and the rate of inhibition of adhesion was expressed as a percentage, with the amount of insoluble glucan attached in a control to which no plant extract was added as 100%. In addition, the solvent used to extract each plant is listed in parentheses in the table. Note that for the control for the aqueous extract of the plant, water was added instead of the aqueous extract of the plant, and for the control for the danol extract of the plant, ethanol was added instead of the ethanol extract of the plant.

1) Production of solvent extract: Purified water (3-) or ethanol (1.5 ΩM) was added to 250 μm of dried pulverized plant material, thoroughly stirred, and then immersed for 72 hours (during which time stirring was performed every 24 hours). ). The mixture was then centrifuged at 3000 rpm for 5 minutes, and the supernatant was collected and used as a solvent extract.

2) Preparation of GTF for assay: Streptococcus mutans OMZ 176 strain was cultured in Berman medium (tryptocase peptone (manufactured by BBL) 2% (hereinafter referred to as weight/volume percentage), glucose 0.1%, phosphoric acid Potassium dihydrogen 0.1%, potassium hydrogen phosphate 0
．． 4%, sodium chloride 0. 2%, magnesium sulfate 0.012%, manganese sulfate 0°0015%, I
)H7,4) Inoculated at 300am and incubated at 37℃ in a constant temperature room for 24 hours.
After incubation for an hour, centrifugation was performed at 15,000 g for 20 minutes to obtain a supernatant. This supernatant was subjected to 60% saturated ammonium sulfate salting out (20 hours at room temperature) to precipitate the proteins in the supernatant.
Centrifugation was performed at 00g for 30 minutes to obtain a precipitate. The precipitate was dissolved in 0.01M phosphate buffer (pH 6.5) and dialyzed three times against 0.01M phosphate buffer (pH 6.5) to obtain GTF for assay. And the GT for the test
F was dispensed into small portions, stored frozen at -20°C, and dissolved in water at the time of assay.

3) Adhesion inhibition test: Conducted using a 96-well microplate (made of vinyl chloride). Add O, 1M phosphate buffer (pH 6,0) 204 containing 10% sucrose, solvent extract (concentration 3%) 5- and GTF 25- for assay into the plate,
Leave at 37°C for 16 hours. After removing the reaction solution in the wells, the plate is slowly immersed in physiological saline (1Ω) and washed by shaking 2 to 3 times. 100 d of 0.5N sodium hydroxide was added to each well to dissolve insoluble glucan, and the amount of sugar was measured by the phenol-sulfuric acid method to calculate the amount of attached insoluble glucan. That is, 40 parts of a 0.5N sodium hydroxide solution, 40 parts of purified water, and 2 parts of a 5% phenol solution.
After stirring, add concentrated sulfuric acid and cool with water for 30 minutes.
Absorbance was measured at 490 nm after minutes.

The results are shown in Table 1.

(Margins below) The anti-GTF action of yaw, chinensis, and zhu is extremely effective in inhibiting the formation of dental plaque, and is useful as a preventive agent for dental caries or periodontal disease.

The amount of the enzyme inhibitor containing a solvent extract as an active ingredient in the oral composition of the present invention is not fixed because the effect of the solvent extract as an active ingredient varies depending on the extraction method. in alcohol (for example, if water is the solvent, 1 part by weight of plant to part by weight of water, or if ethyl alcohol is the solvent, 5 parts by weight of ethyl alcohol)
．． When using a solvent extract obtained by immersing 8 parts by weight of plants in 1 part by weight of plants for 72 hours, the total amount of 0.
It is preferably 1 to 5% (by weight, the same hereinafter), particularly 1 to 3%.

As other components of the present invention, appropriate components may be used depending on the type of oral composition. For example, in toothpaste, powdered toothpaste, water toothpaste, mouthwash, and other toothpastes, abrasives, binders, thickeners, foaming agents, fragrances, sweeteners, preservatives, etc. can be added.Ingredients include geraniol denatured alcohol, Heacetylated saccharide denatured alcohol, phenylethyl alcohol denatured alcohol, brucine denatured alcohol, linalool denatured alcohol, diethyl phthalate denatured alcohol, linaryl acetate denatured alcohol, benzyl acetate denatured alcohol, 10% denatonium benzoate alcohol solution denatured alcohol, Sodium N-acyl-L-glutamate, ascorbic acid, allantoin chlorhydroxyaluminum, sodium alginate,
Aloe extract, benzoic acid, sodium benzoate, titanium mica, ethanol, ethylenediaminetetrapolyoxyethylenepolyoxybrobylene, edetate disodium, edetate tetrasodium dihydrate, zinc chloride, sodium chloride, alkyldiaminoethylglycine hydrochloride, Chlorhexidine hydrochloride, pyridoxine hydrochloride, eugenol, scutellariae extract, γ-oryzanol, kaolin, brown algae extract, carrageenan, carboxyvinyl polymer, sodium carboxymethyl cellulose, hydrated amorphous silicon oxide, licorice extract, dried aluminum hydroxide gel, agar, dilute hydrochloric acid, xanthan gum , guaiazulene, citric acid, sodium citrate, glycerin, glycyrrhizic acid and its salts, chlorhexidine gluconate, gunjo, aluminum magnesium silicate, sodium silicate, light calcium carbonate, cinnamic alcohol, cinnamic aldehyde, crystalline cellulose, N- Hydrogenated beef tallow fatty acid acyl-L-sodium glutamate, synthetic sodium/magnesium silicate, wheat germ oil, saccharin, sodium saccharin, methyl salicylate, aluminum oxide, titanium oxide, Sikon extract, ground calcium carbonate, tartaric acid, distilled peppermint water, sucrose fatty acid Ester, new oil type glyceryl monostearate, aluminum hydroxide, sodium hydroxide, sodium N-stearoyl-L-glutamate, stevia extract, spearmint oil, purified water,
Sage extract, sodium cetyl sulfate, soap base,
Gelatin, sodium fibrin glycolate, sorbitol, potassium sorbate, talc, sodium bicarbonate,
Thymol, sodium dehydroacetate, sodium copper chlorophyllin, sodium lactate, lactose, glycerin, white sugar, peppermint oil, calcium hydrogen phosphate for toothpaste, isobutyl paraoxybenzoate, isopropyl paraoxybenzoate, ethyl paraoxybenzoate, butyl paraoxybenzoate, Propyl paraoxybenzoate, methyl paraoxybenzoate, hydroxyethyl cellulose, calcium pyrophosphate, 1,3-butylene gelicol, glucose, propatool, propylene glycol, bentonite, sodium polyacrylate, polyethylene glycol 30o
, polyethylene glycol 400, polyethylene glycol 1500, polyethylene glycol 6000, polyoxyethylene hydrogenated castor oil, polyoxyethylene stearyl ether, polyoxyethylene cetyl ether,
Polyoxyethylene (200) polyoxypropylene glycol (70), polyvinylpyrrolidone, sodium polyphosphate, d-borneol, microcrystalline wax, maltitol, beeswax, myristic acid jetanolamide, anhydrous ethanol, anhydrous citric acid, silicic anhydride , anhydrous aluminum silicate, anhydrous sodium pyrophosphate, methyl cellulose, ■-menthol, dl-menthol, sorbitan monooleate, polyoxyethylene sorbitan monooleate (20 mol addition), polyoxyethylene sorbitan monopalmitate, (20 mol) addition), sorbitan monolaurate, polyoxyethylene sorbitan monolaurate (20 mole addition), coconut oil fatty acid jetanolamide, eucalyptus oil, sodium lauryl sulfate, jetanolamide lauryl sulfate, sodium lauroyl sarcosinate, liquid paraffin, phosphoric acid Examples include sodium hydrogen phosphate, sodium dihydrogen phosphate, trisodium phosphate, calcium hydrogen phosphate, magnesium phosphate, and fragrance. In addition, troches, liniments, tablets, powders, mouthwashes, mouth fresheners, chewing gum,
Even in drinks, candies, aerosols, etc., ingredients are appropriately blended according to the properties of the product.

In the present invention, in addition to the above-mentioned plant solvent extract, dextranase, lytic enzyme (lysozyme chloride, etc.), mutanase 1, sorbic acid, alexidine, hinokitiol, cetylpyridinium chloride, alkylglycine, alkyldiaminoethyl Glycine salt, allantoin, β-aminocaproic acid, tranexamic acid, azulene, vitamin E, sodium monofluorophosphate, sodium fluoride, cuprous fluoride, water-soluble mono- or di-phosphate, quaternary ammonium compound It is also possible to incorporate active ingredients such as. Other herbal medicine extracts include sagebrush, oren, ryokyu, perilla, kashu, kuromoji, nantenjitsu, hawthorn, botanhi,
Extracts of koju, chinensis, chinensis, chinensis, perilla, ginseng, Japanese rhubarb, rhubarb, red wattle, fenugreek, bellflower, fenugreek, oak, chinensis, clove, acacia japonica, sycamore, and the like may also be used.

<Function> The oral composition according to the present invention contains one or more solvent extracts of plants selected from the above-mentioned peony, chinensis, chinensis, and chinensis, thereby inhibiting GTF derived from Streptococcus mutans. It effectively suppresses the formation of plaque, thus effectively preventing the formation of major surfaces and periodontal disease.

<Example> The composition of the present invention will be specifically explained with reference to Examples below. However, the present invention is not limited to the examples.

Example 1 The following formulations were mixed to obtain a Streptomyces brush containing the Chuyu solvent extract of the present invention.

Dibasic calcium phosphate dihydrate 50.0% glycerin 18.0% carboxymethylcellulose 1.0% sodium lauryl sulfate 10% fragrance
1.0% saccharin
01% Chuyu (50% ethyl alcohol solution extract)
3.0% sodium fluoride 0.1% purified water
Residue 100.0% Example 2 The following formulations were mixed to obtain a water toothpaste containing the chinensis solvent extract of the present invention.

Ethanol 148% Glycerin 100% Polyoxyethylene (60) Hydrogenated Castor Oil 0.4% Sodium Monohydrogen Phosphate 002% Phosphoric Acid 2
Sodium hydrogen 001% Sodium saccharin 0.003% Butyl paraoxybenzoate 0.007% Diyouinthine (extracted with ethyl alcohol)
1.0% dye
0.2% purified water
100.0% Example 3 The following formulations were mixed to obtain a water toothpaste containing the peony solvent extract of the present invention.

Ethanol 5.0% Sorbitol 70% aqueous solution 10.0% Sodium lauryl sulfate
1.0% saccharin sodium 0
05% Butyl paraoxybenzoate 0.05% Peony (water extraction) 2.0% Fragrance -0.2% Purified water Appropriate 100.0% Example 4 The following formulations were mixed, and the solvent extraction of the present invention was performed. A mouthwash containing the product was obtained.

Ethanol 20.0% Glycerin 15.0% Polyoxyethylene/polyoxypropylene polymer 1, O% Sodium saccharin 0.02% Butyl paraoxybenzoate 0.05% Diyointhine (ethanol extract) 3.0% Color
Trace fragrance 0
．． 2% purified water suitable 1
00.0% Example 5 The following formulations were mixed to obtain a candy containing the peony solvent extract of the present invention.

Granulated sugar 46.3% starch syrup
36.7% citric acid
1. 6% Peony (water extract) 1.0
% Fragrance 0. 1% water
Compatibility: 100.0% Example 6 The following formulations were mixed to obtain a candy containing the loquat extract of the present invention.

Reduced malt starch syrup 90.0% citric acid
1.6% Biwayo (water extract) 1.
0% Fragrance 0. 1% water
Continuous 100.0% Example 7 The following formulations were mixed and filled into an aerosol can with a propellant to obtain a mouth spray containing the solvent extract of chinensis of the present invention.

Ethanol (90 proof)
41.75% deionized water 41.75%
Sorbitol 10.00% Glycerin 3.00% POE Sorbitan Diisostearate 1. 50
% Diyointin (Ethanol extraction 1) 1.
00% Sodium Saccharin 0.50% Example 8 The following formulations were mixed to obtain a dental cream containing the peony solvent extract of the present invention.

Sorbitol 42.45% glycerin
24.00% Cyropunto 704 (silica-containing polish) 22.0 0% deionized water 3.00% polyethylene glycol 600 3,00% Sorbitan diisostearate
1. 50% titanium dioxide 0.
50% Sodium Benzoate 0.50% Sodium Carboxymethyl Cellulose
0. 35% sodium saccharin 1.70
% Peony (water extract) 1.00% <Effects of the Invention> The oral composition according to the present invention is made from one or more solvent extracts of plants selected from the above-mentioned Peony, Biwayou, Jyoinchin, and Chiyu. By containing an enzyme inhibitor, G. mutans derived from Streptococcus mutans
The formation of dental plaque by TF inhibition is effectively suppressed, and therefore the occurrence of dental caries and periodontal diseases are effectively prevented.

(that's all) Patent applicant: Earth Pharmaceutical Co., Ltd.

Claims (2)

[Claims] (1) An enzyme inhibitor characterized by containing one or more solvent extracts of plants selected from peonies, peonies, peonies, and chiyus. (2) An oral composition characterized by containing the above-mentioned enzyme inhibitor as an active ingredient.