KR20150118144A - Oral composition - Google Patents

Abstract

(A) human dipenin and (B) ascorbic acid, which are effective in preventing or treating periodontal disease, and which are excellent in the bactericidal effect against oral bacteria, especially the periodontal disease causative microorganism Porphyromonas jinja valis, Or an acid anhydride or a salt thereof.

Description

Oral composition {ORAL COMPOSITION}

The present invention relates to a method for preventing and treating periodontal disease, which is excellent in the bactericidal effect against oral bacteria, especially Porphyromonas japonica, which is a cause of periodontal disease, and the cytotoxic effect in oral cavity, The present invention relates to an oral composition containing diphencin.

The periodontal disease is an infection caused by a bacterium mainly consisting of anaerobic gram-negative bacteria such as Porphyromonas jingivalivalis. It is an extermal toxin such as leukotoxin produced by bacteria and endotoxin (lipopolysaccharide) Inflammation is induced, and tissue is damaged. On the other hand, in the living body, neutrophils and lymphocytes infiltrate (infiltrate) the periodontal pocket or the gum tissue, and bacteria are phagocytosed, and a specific antibody is generated to exclude these foreign bodies It is pointed out that the excessively damaged active oxygen in the phagocytosis further damages the living tissue. For example, when a gut-derived fibroblast cell is injured by reactive oxygen species, it causes collagen fiber breakdown and cell proliferative capacity deterioration, so that the gums retract (retract) Go ahead.

Therefore, it is considered effective to inhibit cytotoxicity in the oral cavity as well as sterilization of pathogenic bacteria causing periodontal disease in the oral cavity for prevention and treatment of periodontal disease such as periodontitis, and to provide an oral cavity composition having both effects Is required.

Regarding sterilization of periodontal pathogenic bacteria in oral cavity, it is known that human dipentin, a peptide derived from a host, has antibacterial action against a wide range of oral pathogenic bacteria and is safe for human body. Patent Document 1 (Japanese Unexamined Patent Publication (Kokai) No. 2001-288105) discloses that human β-diphencine has a strong antimicrobial activity against Actinobacillus actinomycetemcomitans, a causative organism of periodontitis, and Porphyromonas gingivalis, and the like have been proposed, and a human dipenin-containing skirt has been proposed. In addition, Patent Document 2 (Japanese Patent Laid-Open Publication No. 2009-155214) proposes an oral composition in which an antibacterial effect of a biofilm in the oral cavity is improved by using human diffenzin and dextranase in combination.

However, it is difficult to say that human dyspensin has a sufficient sterilizing effect on periodontal pathogenic bacteria, and improvement of sterilizing power is a problem.

On the other hand, regarding the inhibition of cytotoxicity in the oral cavity, ascorbic acid ester or its salt such as ascorbic acid phosphate ester or the like salt is used to protect the living tissue from oxidative stress injury by dissolving excessive active oxygen produced in the living body, And has an effect of inhibiting the injury of cells in the oral cavity. However, the cytotoxic inhibitory effect still has room for improvement. In addition, the ascorbic acid ester or a salt thereof does not have a sterilizing effect on the causative bacteria of periodontal disease.

Japanese Patent Application Laid-Open No. 2001-288105 Japanese Patent Application Laid-Open No. 2009-155214

The present invention has been made in view of the above circumstances, and it is an object of the present invention to provide an oral composition which is excellent in the germicidal effect against bacteria in the oral cavity, in particular, the causative bacteria of periodontal disease and the cytotoxic effect in the oral cavity, and is effective for prevention and treatment of periodontal disease The purpose.

As a result of diligent studies to achieve the above object, the inventors of the present invention have found that, when (A) human diphenzin and (B) ascorbic acid ester or a salt thereof are used in combination, oral bacteria, particularly, (Hereinafter, abbreviated as Pg bacterium), and the effect of inhibiting the injury to the oral cavity were remarkably improved.

That is, human dipentin, which is a peptide derived from a host, has antimicrobial activity against periodontal pathogens and is known as a safe ingredient for human body. However, the inventors of the present invention have studied about the bactericidal effect thereof, There is a problem that there is no sterilization effect. As a result of further studies to solve this problem, it has been found that (A) a combination of human diffpin and (B) ascorbic acid ester or a salt thereof, especially ascorbic acid phosphoric acid ester, ascorbic acid sulfuric acid ester or a salt thereof , The components (A) and (B) act synergistically and synergistically, which enhances the bactericidal effect in the oral cavity derived from human diphenzene, particularly the bactericidal effect against the causative bacteria of periodontal disease, Ester, or a salt thereof, which is effective in inhibiting oxidative stress caused by inhibition of oxidative stress, thereby achieving a superior bactericidal effect in oral cavity and an effect of inhibiting cytotoxicity. In addition, by incorporating the combination system of the components (A) and (B) into the composition for oral cavity, it is possible to obtain a preparation effective for prevention and treatment of periodontal disease, achieving a high bactericidal effect in oral cavity and a cytotoxic inhibitory effect.

In this case, as shown in a comparative example to be described later, the bacteriostatic effect of human diphenzin in the oral cavity was not sufficient, the cytotoxic effect was not recognized, and the cytotoxic inhibitory effect of ascorbic acid ester or its salt was sufficient (A) or (B) is used in combination with the components (A) and (B) in the present invention, The bactericidal effect and the cytotoxic inhibitory effect in the oral cavity could be enhanced. The above-mentioned action and effect of the present invention can be obtained by using histatin, a saliva protein having antibacterial action, which is specific to the combination system of components (A) and (B), instead of component (A) And is not particularly obtained when an ascorbic acid salt such as sodium is used instead of the component (B).

Accordingly, the present invention provides the oral composition described below.

[One]

(B) an ascorbic acid ester or a salt thereof, (A) human diphenzin, and (B) an ascorbic acid ester or a salt thereof.

[2]

The composition for oral use according to [1], wherein the component (A) is human diphenx-β-2 or human dipenin β-3.

[3]

The oral composition according to [1] or [2], wherein the component (B) is at least one selected from ascorbic acid phosphate ester and its salt, ascorbic acid sulfate and salts thereof.

[4]

Wherein the component (B) is ascorbic acid phosphate ester magnesium and / or ascorbic acid phosphate ester sodium.

[5]

The oral composition according to any one of [1] to [4], wherein the composition contains 0.000005 to 0.0005 mass% of component (A) and 0.1 to 2 mass% of component (B).

[6]

The oral composition according to any one of [1] to [5], wherein the component (B) is in a mass ratio of 200 to 400,000 with respect to the component (A).

[7]

The oral composition according to any one of [1] to [6], which is a skirt or a remedy.

[8]

The oral composition according to any one of [1] to [7], which is for preventing or treating periodontal disease.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a human diphenocin-containing bacterium which is excellent in the germicidal effect against oral bacteria, particularly Pg bacteria causing periodontal disease and the cytotoxic effect in the oral cavity, Oral compositions may be provided.

Hereinafter, the present invention will be described in more detail. The oral composition of the present invention is characterized by (A) human diphenzin and (B) ascorbic acid ester or a salt thereof in combination, and by such a combination system, the oral bactericidal effect and The injury inhibiting effect is improved.

(A) Human dyspensin includes human dyspensin [alpha], human dyspensin [beta] -l, human dyspensin [beta] -2 and human dyspensin [ Two or more of them can be used, and in particular, human dyspensin β-2 and human diphenzin β-3 are preferable in terms of bactericidal effect of periodontal disease bacteria.

Commercially available human dyspensin can be used, for example, human dyspensin β-2 manufactured by Peptide Institute and human dyspensin β-3 manufactured by Anygen.

The blending amount of (A) human diffizine is preferably 0.000005 to 0.0005% (mass%, the same applies hereinafter), more preferably 0.00005 to 0.0005% of the whole composition. As the compounding amount increases, the bactericidal effect in the oral cavity and the cytotoxic inhibitory effect are improved, and 0.000005% or more compounding is suitable for manifesting the effect. If the blending amount is too large, the effect can not be improved already, and the feeling of use may be lowered due to the appearance of an unusual taste, so that the content is preferably 0.0005% or less.

The ascorbic acid ester which is the component (B) is one in which one or two or more hydroxyl groups (hydroxyl groups) at any of positions 2, 3, 5 and 6 of ascorbic acid are phosphoric acid, polyphosphoric acid, sulfuric acid, Or other pharmacologically acceptable compounds.

As ascorbic acid ester or a salt thereof, particularly preferred is ascorbic acid phosphoric acid ester or salt thereof, ascorbic acid sulfuric acid ester or a salt thereof. Ascorbic acid phosphoric acid ester and ascorbic acid sulfuric acid ester are those in which one or two or more of the hydroxyl groups at positions 2, 3, 5 and 6 of ascorbic acid are phosphoric acid compounds such as phosphoric acid, polyphosphoric acid, Specific examples thereof include ascorbic acid-2-phosphate ester, ascorbic acid-3-phosphate ester, ascorbic acid-6-phosphate ester, ascorbic acid-2-polyphosphoric acid ester, ascorbic acid- Sulfuric acid esters. Examples of the salt include alkali metal salts such as sodium salt, potassium salt, calcium salt and magnesium salt, and alkaline earth metal salts. As the component (B), one or more selected from these may be used. Particularly, from the viewpoint of the effect of preventing oral bioflavonemia, particularly the effect of inhibiting cytotoxicity, the magnesium salt of ascorbic acid phosphate ester , And sodium salts are suitably used.

The amount of the ascorbic acid ester (B) or its salt is preferably from 0.1 to 2%, more preferably from 0.2 to 1%, based on the whole composition. As the blending amount increases, the oral bacterial bactericidal effect and cytotoxic inhibitory effect are improved, and blending of 0.1% or more is suitable for improvement of oral bacterial bactericidal effect and cytotoxic inhibitory effect. In addition, it is suitable to improve the germ bactericidal effect in the oral cavity and to secure a good feeling of use when it is 2% or less. If too much is added, the bactericidal effect in the oral cavity may be lowered. In addition, there is a case where a feeling of use is lowered due to the appearance of a different taste.

In the present invention, it is preferable that the ascorbic acid ester (B) or its salt (A) is 20 times or more, especially 200 to 400,000 times, more preferably 400 to 200,000 times, more preferably Preferably 1,000 to 10,000,000 times, particularly preferably 1,000 to 10,000 times. When the blending ratio is in this range, the effect of the present invention is further improved.

The oral composition of the present invention can be used as a dentifrice such as a lotion, a liquid skim, a liquid skim, a lubricious skim, , And a mouth rinse, but it is particularly suitable for a skirt preparation. In this case, in accordance with the purpose of the composition, the formulation, etc., well-known components other than the above-mentioned components may be compounded within a range not hindering the effect of the present invention. For example, as a skimming agent, an abrasive, a tackifier, a binder, a surfactant, and, if necessary, a perfume, a sweetener, a colorant, an antiseptic, an effective ingredient and the like are mixed.

Examples of the abrasive include silica-based abrasives such as silica gel, precipitated silica, aluminosilicate, and zirconosilicate, calcium secondary dibasic hydrate and anhydride, calcium tertiary phosphate, calcium quaternary phosphate, calcium pyrophosphate, calcium carbonate, aluminum hydroxide , Alumina, magnesium carbonate, magnesium triphosphate, zeolite, hydroxyapatite, synthetic resin abrasive, and the like. The blending amount of the abrasive is adjusted according to the formulation, and it is preferably 2 to 40%, particularly 10 to 30% in a soft skirt, and 0% in a liquid skirt.

Examples of the viscosifier include sugar alcohols such as sorbitol, xylitol, maltitol and lactitol, and polyhydric alcohols such as glycerin, propylene glycol and polyethylene glycol. The blending amount is usually 5 to 50%, particularly 20 to 45% desirable.

Examples of the binder include cellulose derivatives such as carboxymethylcellulose sodium, methylcellulose and hydroxyethylcellulose, rubbers such as xanthan gum and gum arabic, organic dispersants such as carrageenan, polyvinyl alcohol and sodium polyacrylate, , Gelling aluminum oxide, non-rubber, and laponite. The blending amount is usually 0.1 to 5% in the case of a plain skirt, and 0 to 5% in the case of a liquid skirt or a cleanser.

As the surfactant, anionic surfactant, nonionic surfactant, cationic surfactant, and amphoteric surfactant can be mixed. have.

Examples of the anionic surfactant include alkyl sulfates such as sodium lauryl sulfate, N-acyl sarcosinates such as N-lauroyl sarcosine sodium and N-myristoyl sarcosinate, N -Acyl glutamate, sodium N-methyl-N-acyltaurate, sodium N-methyl-N-acylalanine, sodium -olefinsulfonate and the like.

Examples of the nonionic surfactant include sugar alcohol esters such as sucrose fatty acid esters and maltose fatty acid esters, sugar alcohol fatty acid esters such as maltitol fatty acid esters and lactitol fatty acid esters, sorbitan fatty acid esters, glycerin fatty acid esters, Polyglycerin fatty acid esters such as glyceryl monolaurate, decyl glyceryl monolaurate and decaglyceryl monolaurate, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monostearate and the like Polyoxyethylene fatty acid esters such as polyoxyethylene sorbitan fatty acid esters and polyoxyethylene hardened castor oil, polyoxyethylene higher alcohol ethers such as polyoxyethylene lauryl ether, fatty acid alkanol amides such as lauric acid diethanolamide, Polyoxyethylene polyoxypropylene copolymerization Polyoxyethylene polyoxypropylene fatty acid esters, and the like.

Examples of the cationic surfactant include alkylammonium and alkylbenzylammonium salts. Examples of the amphoteric surfactant include betaine such as alkyl betaine, fatty acid amide propyl betaine and alkyl imidazolinium betaine.

The blending amount of the surfactant is preferably from 0.001 to 10%, particularly preferably from 0.1 to 5%, of the whole composition.

Examples of the perfume include perfume oils such as peppermint oil, spearmint oil, anise oil, eucalyptus oil, wintergreen oil, cassia oil, kurobu oil, time oil, sage oil, lemon oil, orange oil, peppermint oil, cardamon oil, Lavender oil, Rosemary oil, Laurel oil, Camomile oil, Caraway oil, Marjoram oil, Bayou, Lemon glass oil, Oriental oil, Pine needle oil, Neroli oil, Rose oil, Jasmine oil, Grapefruit oil, Sweet oil, Natural fragrances such as iris concrete, absorptive peppermint, absorro rose and orange flower, and processing of these natural fragrances (such as a pre-cut, a wake cut, sorting, liquid extraction, Perfume and the like) and flavorings such as menthol, carboen, anethole, cineol, methyl salicylate, cinnamic aldehyde, ogenol, 3-l-mangthoxypropane-l, 2-diol, , Linalyl acetate, limonene, menton, menthyl acetate, N-substituted-para Carboxylacetate, anisaldehyde, ethyl acetate, ethyl butyrate, allyl cyclohexane propionate, methyl anthranilate, ethyl methylphenyl glycidate , Vanillin, undecalactone, hexanalbutanol, isoamyl alcohol, heptanol dimethylsulfide cyclotene, furfural, trimethylpyrazine, ethyl lactate, ethyl thioacetate, and the like, as well as Strawberry flavor, Known fragrance materials used in oral compositions, such as a combination flavor such as a flavor, a banana flavor, a pineapple flavor, a grapeflavor, a mango flavor, a butter flavor, a milk flavor, a flavor mix flavor and a tropical flavor flavor.

The blending amount is not particularly limited, but it is preferable that the above-mentioned perfume material is used in an amount of 0.000001 to 1% in the formulation. It is preferable that 0.1 to 2% of the fragrance ingredient be used as the fragrance ingredient for an intention flavor.

Examples of the sweetener include saccharin sodium, stevioside, paramethoxycinnamic aldehyde, and perylartin. Examples of the colorant include Blue No. 1, Yellow No. 4, and titanium dioxide.

Examples of the preservative include benzoic acid or salts thereof such as paraoxybenzoic acid ester and sodium benzoate.

Examples of the active ingredient include non-human diphenzenes and ascorbic acid esters and salts thereof, nonionic bactericides such as isopropylmethylphenol, catechins such as cetylpyridinium chloride, chlorhexidine hydrochloride, benzalkonium chloride, An enzyme such as dextrinase, mitanase, amylase, protease, sodium fluoride, sodium monofluorophosphate, etc. may be used as an anti-inflammatory agent such as an anti-inflammatory agent, A water-soluble phosphoric acid compound such as a potassium salt and a sodium salt of polyphosphoric acid, a copper compound such as copper gluconate and sodium copper chlorophyllin, an inorganic salt such as sodium chloride, potassium nitrate, aluminum lactate, zinc chloride, zinc citrate, Vitamins such as tocopherol acetate, zeolite, azulene, dihydrocholesterol, chlorophyll, Angelica koreana extract, time, It may include gold, sperm, plant extracts such as witch hazel, tartar agents, anti-plaque, etc. These effective ingredients can be blended in an effective amount within a range not hindering the effect of the present invention.

Example

Hereinafter, the present invention will be described in detail by way of examples, comparative examples, and formulations, but the present invention is not limited to the following examples. In the following examples,% represents mass% unless otherwise specified.

[Practical example, comparative example]

A sample solution having the composition shown in Table 1 was prepared by a routine method, and the cytotoxic inhibitory effect and bactericidal effect in the oral cavity were evaluated by the methods shown in the following experimental examples. The results are listed in the table.

<Experimental Example 1> Test for inhibiting cytotoxicity

(D-MEM) containing 10% fetal bovine serum (FBS) at 37 ° C under a condition of 5% CO _{2 in} the presence of a commercially available gum fibroblast gin-1 (DS Pharma Biomedical) And then pre-cultured. Gin-1, which had been prepared at 5 × 10 ⁴ cells / ml, was inoculated on a 48-well plate at 400 μl and cultured for another 24 hours. After removing the culture solution, 400 μl each of the sample solutions shown in the table was added (diluted 20-fold with D-MEM containing 10% FBS) and treated for 48 hours with the drug.

After completion of the treatment, 1 mM aqueous hydrogen peroxide solution (supposing active oxygen) was added for 60 minutes. After completion of the treatment, the sample solution was removed, and 400 μl of a solution containing 1 mM hydrogen peroxide (D-MEM containing 10% FBS) was added and treated for 60 minutes. After the solution was removed, 200 쨉 l of a cell activation reagent (Calcein AM; manufactured by Invitrogen) was added, and the mixture was incubated at 37 째 C under 5% CO ₂ for 30 minutes. Thereafter, the fluorescence intensity was measured under the condition of Ex / Em = 485 nm / 538 nm using a plate reader (Fluoroskan Ascent, manufactured by Labsystems).

The result calculated from the following equation (1) was used as a cytotoxic inhibition rate. The cytotoxicity inhibition test was carried out at n = 10. The average value was calculated, and the cytotoxic inhibitory effect was evaluated based on the following criteria.

(%) = (Fluorescence intensity in the case of hydrogen peroxide treatment after sample solution treatment) / (fluorescence intensity in case of no hydrogen peroxide treatment after sample solution treatment) 占 100 (One)

Rating criteria for cytotoxic inhibitory effect;

?: Cytotoxic inhibition rate is 90% or more

○: the cytotoxic inhibition rate is 70% or more and less than 90%

?: Cytotoxic inhibition rate is 50% or more and less than 70%

X: Less than 50% inhibition of cytotoxicity

<Experimental Example 2> Bactericidal effect test in oral cavity

(Manufactured by Wako Pure Chemical Industries, Ltd., 500 占 퐂 / ml) and Menadione (100 占 퐂 / ml, manufactured by Wako Pure Chemical Industries, Ltd.) A suspension of bacteria was prepared so as to have a concentration of 1 x 10 ⁹ cells / ml of physiological saline solution of Porphyromonas jinja valis ATCC33277 Co., Ltd. The bacterial solution and the sample solution indicated in the table were mixed at a ratio of 1: 1 (volume ratio) and left to stand for 3 minutes. Thereafter, 4-step dilution was performed with physiological saline 10-fold, and the plate was plated on a blood agar plate with a spiral platter. As a control, a physiological saline solution and a bacterial solution were mixed at a ratio of 1: 1 (volume ratio) instead of the drug.

Blood plates were incubated for 5 days under anaerobic conditions (95 vol% nitrogen, 5 vol% carbon dioxide) at 37 DEG C, and the number of colonies formed was measured. This oral bacterial sterilization test was carried out at n = 5. The average value was calculated, and the number of colonies of the sample solution relative to the number of colonies of the control solution was determined as the viable cell count, and the bactericidal effect in the oral cavity was evaluated based on the following criteria.

Evaluation criteria for oral bacterial bactericidal effect;

&Amp; cir &amp;: The viable cell ratio was less than 10%

○: Compared with the control, the viable cell ratio is less than 10% to 40%

[Delta]: 40% to less than 80% of live cells compared with the control

X: the viable cell ratio was 80% or more as compared with the control

Details of the raw materials used are shown below.

(A) Human dipencin (human dipentin beta-2, manufactured by Peptide Research Institute)

(B) Ascorbic acid-2-phosphate ester magnesium (manufactured by Wako Pure Chemical Industries, Ltd.)

(B) Sodium ascorbyl-2-phosphate ester (manufactured by Wako Pure Chemical Industries, Ltd.)

(B) Sodium ascorbic acid-2-sulfate (manufactured by Wako Pure Chemical Industries, Ltd.)

Sodium ascorbate (manufactured by Wako Pure Chemical Industries, Ltd.)

Histatin 5 (manufactured by Wako Pure Chemical Industries, Ltd.)

[Table 1]

Hereinafter, a prescription example is shown. All of the following examples have excellent cytotoxic effect and bactericidal effect in the oral cavity.

[Prescription Example 1] Open skirt

(A) Human depensin β-2 (manufactured by Peptide Research Institute) 0.00001%

(B) ascorbic acid-2-phosphate ester magnesium

 (Manufactured by Wako Pure Chemical Industries, Ltd.) 0.5

Anhydrous silicic acid 20.0

Sodium lauryl sulfate 1.0

Propylene glycol 3.0

Sour bit liquid (100% product) 30.0

Xanthan Rubber 1.0

Sodium saccharin 0.1

Spice 1.0

Purified water                                   balance

Total 100.0%

[Prescription Example 2] Liquid skirt

(A) 0. &lt; RTI ID = 0.0 &gt; 5% &lt;

(B) ascorbic acid-2-phosphate ester magnesium

 (Manufactured by Wako Pure Chemical Industries, Ltd.) 0.5

Xylitol 3.0

Glycerin (AI = 100) 2.0

Polyoxyethylene (60) Hardened castor oil (HCO-60) 0.5

Sodium citrate 0.3

Citric acid 0.1

Sodium benzoate 0.3

Benzoic acid methyl 0.1

Fragrance 0.2

Saccharin sodium 0.002

Propylene glycol 3.0

Polyethylene glycol 5.0

Purified water                                   balance

Total 100.0%

[Prescription Example 3] Liquid skirt

(A) 0. &lt; RTI ID = 0.0 &gt; 01% &lt;

(B) ascorbic acid-2-phosphate ester magnesium

 (Manufactured by Wako Pure Chemical Industries, Ltd.) 0.5

Xylitol 3.0

Glycerin (100% product) 5.0

Polyoxyethylene (60) Hardened castor oil (HCO-60) 0.5

Sodium citrate 0.3

Citric acid 0.1

Fragrance 0.2

Ethanol 8.0

Propylene glycol 3.0

Purified water                                   balance

Total 100.0%

Claims (8)

(B) an ascorbic acid ester or a salt thereof, (A) human diphenzin, and (B) an ascorbic acid ester or a salt thereof. The method according to claim 1,
Wherein the component (A) is human depensin [beta] -2 or human depensin [beta] -3. 3. The method according to claim 1 or 2,
Wherein the component (B) is at least one selected from ascorbic acid phosphate ester and its salt, ascorbic acid sulfuric acid ester and salts thereof. The method of claim 3,
Wherein the component (B) is magnesium ascorbyl phosphate ester and / or sodium ascorbyl phosphate ester. 5. The method according to any one of claims 1 to 4,
(A) in an amount of 0.000005 to 0.0005 mass%, and (B) in an amount of 0.1 to 2 mass%. 6. The method according to any one of claims 1 to 5,
Wherein the component (B) is in a mass ratio of 200 to 400,000 times with respect to the component (A). 7. The method according to any one of claims 1 to 6,
Wherein the oral composition is a cosmetic or dermatological agent. 8. The method according to any one of claims 1 to 7,
Wherein the composition is for preventing or treating periodontal disease.