WO2014125975A1

WO2014125975A1 - Oral composition

Info

Publication number: WO2014125975A1
Application number: PCT/JP2014/052671
Authority: WO
Inventors: 貴士近澤; 眞里阿部; 苗穂鈴木
Original assignee: ライオン株式会社
Priority date: 2013-02-14
Filing date: 2014-02-05
Publication date: 2014-08-21
Also published as: JP6222216B2; JPWO2014125975A1

Abstract

Provided is an oral composition which has an excellent cytotoxicity inhibitory effect in the oral cavity, and an excellent bactericidal effect against oral bacteria, in particular, porphyromonas gingivalis, a periodontal disease-causing bacteria, and which is effective in the prevention and treatment of peridontal disease. This oral composition is characterized by containing (A) ascorbic acid ester or a salt thereof, and (B) a methylene blue.

Description

Oral composition

The present invention includes an ascorbic acid ester or a salt thereof, which has an excellent cytostatic effect and a bactericidal effect on oral bacteria, particularly porphyromonas gingivalis, which is a causative agent of periodontal disease, and is effective in preventing and treating periodontal disease It relates to the composition for oral cavity.

Periodontal disease is an infection caused by bacteria, mainly anaerobic gram-negative bacteria such as Porphyromonas gingivalis, etc., and exotoxins (leucotoxins etc.) and endotoxins (lipopolysaccharides etc.) produced by the bacteria ) Induces inflammation and damages the tissue. On the other hand, in the living body, neutrophils and lymphocytes infiltrate the periodontal pockets and gingival tissues, phagocytosing bacteria, and creating an immune response that eliminates these foreign substances by creating specific antibodies, In recent years, it has been pointed out that active oxygen generated excessively during phagocytosis further damages biological tissues. For example, when fibroblasts constituting the gingiva are injured by active oxygen, collagen fibers are destroyed and the cell growth ability is reduced, so that the gingiva is retracted and periodontal disease progresses.

Therefore, sterilizing oral bacteria, particularly periodontal disease-causing bacteria, and inhibiting oral cell damage as described above is effective for the prevention and treatment of periodontal diseases such as periodontitis. Conceivable. For this reason, development of the technique which provides the composition for oral cavity which satisfy | fills the bactericidal effect in a buccal cavity and a cytotoxic effect simultaneously is desired.

Ascorbic acid esters such as ascorbic acid phosphates or salts thereof are more stable than ascorbic acid salts and protect excess tissues from oxidative stress by erasing excess active oxygen produced in the body. In addition, the effect of suppressing inflammation has been reported. Such an oral composition containing an ascorbic acid derivative is considered to reduce the cytotoxicity by suppressing oxidative stress and exert an effect of preventing periodontal disease. By using an ascorbic acid derivative and arabitol in combination, oxidative stress Patent Document 1 (Japanese Patent Application Laid-Open No. 2012-180330) proposes that the suppression effect is improved and the effect continues for a long time.
However, the cytotoxic effect of ascorbic acid ester or its salt still has room for improvement, and ascorbic acid ester or its salt itself has no bactericidal action against oral bacteria.

On the other hand, methylene blue, which is generally used as a dye or dyeing liquid and is known as a kind of pigment, has recently been attracting attention because it is considered to exhibit a bactericidal and disinfecting action by an oxidation-reduction action. Patent Document 2 (Japanese Patent No. 5041617) proposes that when a compound that gives zinc ions and methylene blue are used in combination, they have a bactericidal action against oral bacteria.

However, methylene blue is not commonly used as a component of oral compositions and is rarely used, and the sterilizing action is still under study.

JP 2012-180330 A Japanese Patent No. 5041617

The present invention has been made in view of the above circumstances, and is excellent in bacteriostatic effects on oral cell damage and bactericidal effects on oral bacteria, in particular, bacteria causing periodontal disease, and is effective for prevention and treatment of periodontal disease. An object is to provide a composition.

As a result of intensive studies to achieve the above object, the present inventors have found that when (A) an ascorbic acid ester or a salt thereof and (B) methylene blue are used in combination, It has been found that the bactericidal effect against bacteria, in particular Porphyromonas gingivalis (hereinafter abbreviated as Pg bacteria), which is a periodontal disease-causing bacterium, has been remarkably improved.

That is, in the present invention, when (A) one or more selected from ascorbic acid esters or salts thereof, particularly ascorbic acid phosphate esters, ascorbic acid sulfate esters and salts thereof, and (B) methylene blue are used in combination, it is surprisingly surprising. The (A) and (B) components act specifically and synergistically, the oxidative stress inhibitory effect derived from ascorbic acid ester or its salt is enhanced, and the oral cytotoxicity inhibitory effect is enhanced. The bactericidal effect of oral bacteria, particularly the above-mentioned periodontal disease-causing bacteria, is enhanced and expressed. Thereby, by blending the above combination system into an oral composition, it is possible to obtain a preparation effective for preventing and treating periodontal disease by simultaneously imparting an excellent cytotoxic effect and a bactericidal effect on oral bacteria. .

In this case, as shown in a comparative example described later, ascorbic acid ester or a salt thereof has a low cytostatic effect and there is room for improvement, and an oral bactericidal effect is not recognized. In spite of the low bactericidal effect of disease-causing bacteria and the absence of cytostatic effect, as shown in the examples, in the present invention, by the specific action of the combined system of the components (A) and (B), (A) or (B) It is possible to provide a significantly higher oral bactericidal effect and cytotoxicity-suppressing effect than the additive effect of the single blending system of the component, and there is a particularly remarkable effect.
Methylene blue is known to have an action to increase the redox potential because it exhibits an oxidation-reduction reaction, and is considered to exhibit a bactericidal disinfection action. However, methylene blue is derived from ascorbic acid ester or a salt thereof. It is a new finding of the present inventors that it contributes to the improvement of the effect of suppressing cytotoxicity and gives the above-mentioned special effects in the oral composition.

Accordingly, the present invention provides the following oral composition.
[1]
An oral composition comprising (A) ascorbic acid ester or a salt thereof and (B) methylene blue.
[2]
(A) The composition for oral cavity as described in [1] whose component is 1 or more types chosen from ascorbic-acid phosphate, ascorbic-acid sulfate, and its salt.
[3]
The composition for oral cavity according to [2], wherein the component (A) is magnesium ascorbate phosphate and / or sodium ascorbate phosphate.
[4]
The composition for oral cavity according to [1], [2] or [3], containing 0.1 to 2% by mass of component (A) and 0.001 to 0.2% by mass of component (B).
[5]
The composition for oral cavity according to any one of [1] to [4], wherein (B) / (A) is from 1/2000 to 1/2 as a mass ratio.
[6]
The oral composition according to any one of [1] to [5], which is a dentifrice or mouthwash.

According to the present invention, the effect of inhibiting oral cell injury and oral bacteria, particularly P. g. An oral composition containing an ascorbic acid ester or a salt thereof, which has an excellent bactericidal effect against bacteria and is effective in preventing and treating periodontal disease, can be provided.

Hereinafter, the present invention will be described in further detail. The composition for oral cavity of the present invention contains (A) ascorbic acid ester or a salt thereof and (B) methylene blue.

Ascorbic acid ester of component (A), one or more of hydroxyl groups at any of positions 2, 3, 5, and 6 of ascorbic acid is phosphoric acid, polyphosphoric acid, sulfuric acid, fatty acid, or other pharmaceutically acceptable. This is an ester of a compound.
As the ascorbic acid ester or a salt thereof, an ascorbic acid phosphate or a salt thereof, or an ascorbic acid sulfate or a salt thereof is particularly preferable. Ascorbic acid phosphoric acid ester and ascorbic acid sulfuric acid ester are phosphoric acid compounds such as phosphoric acid and polyphosphoric acid, or an ester of sulfuric acid. It has become. Specifically, ascorbic acid-2-phosphate, ascorbic acid-3-phosphate, ascorbic acid-6-phosphate, ascorbic acid-2-polyphosphate, ascorbic acid-2-sulfate It is done. 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 (A), one or more selected from these can be used. Particularly, the component is used for the oral cavity, and is an ascorbic acid phosphate ester from the viewpoint of a gingivitis prevention effect, particularly a cytotoxicity suppression effect. The magnesium salt and sodium salt are preferably used.

(A) The blending amount of ascorbic acid ester or a salt thereof is preferably 0.1 to 2% (mass%, the same shall apply hereinafter) of the whole composition, and more preferably 0.2 to 1%. As the compounding amount increases, the cytotoxicity-inhibiting effect and the oral bacteria sterilizing effect are improved. It is preferable to add 0.1% or more to improve the cytotoxicity-inhibiting effect and the oral bacteria-sterilizing effect. Moreover, when there are too many compounding quantities, not only the improvement of an effect cannot be expected any longer, but a taste may appear and a usability | use_condition may fall, It is suitable for ensuring favorable usability | use_condition that it is 2% or less. is there.

(B) Methylene blue is 3,7-bis (dimethylamino) phenothiazinium chloride (IUPAC name), and when used in combination with component (A), enhances the cytotoxic effect of component (A). In addition, P. g. High bactericidal action against bacteria can be imparted. In addition, a commercial item can be used for methylene blue.

(B) The blending amount of methylene blue is preferably 0.001 to 0.2%, more preferably 0.005 to 0.2% of the entire composition. As the compounding amount increases, the cytotoxicity-inhibiting effect and the bactericidal effect in the oral cavity are improved. It is preferable to add 0.001% or more to improve the cytotoxicity-inhibiting effect and the oral bacteria-disinfecting effect. Moreover, when there are too many compounding quantities, not only improvement of an effect cannot be expected any longer, but a nasty taste may be expressed and a feeling of use may fall, and it is suitable for ensuring good feeling of use that it is 0.2% or less. It is.

In the present invention, the ratio (B) / (A)) of the amount of (B) methylene blue to the amount of (A) ascorbic acid ester or salt thereof is 1/2000 or more, particularly 1/2000 as a mass ratio. Is preferably 1/25, more preferably 1/500 to 1/2, and even more preferably 1/200 to 1/2. When methylene blue is blended in such a ratio, the present invention The effect is improved. When (B) / (A) is 1/2000 or more, the cytostatic effect and the bactericidal effect on oral bacteria are particularly improved. It can prevent that the cytotoxic-inhibition effect falls that it is 1/2 or less.

The oral composition of the present invention can be produced by adopting conventional methods for various dosage forms such as toothpaste, toothpaste such as toothpaste, liquid toothpaste, liquid toothpaste, and toothpaste, and mouthwash. Especially suitable for dentifrice. In this case, depending on the purpose of the composition, the dosage form, and the like, appropriate known components can be blended in addition to the above components as long as the effects of the present invention are not hindered. For example, in toothpaste, an abrasive, a thickener, a binder, a surfactant, and, if necessary, a fragrance, a sweetener, a colorant, a preservative, and an active ingredient are blended.

As abrasives, silica-based abrasives such as silica gel, precipitated silica, aluminosilicate, zirconosilicate, dicalcium phosphate dihydrate and anhydrous, tricalcium phosphate, tetracalcium phosphate, calcium pyrophosphate, calcium carbonate, water Examples thereof include aluminum oxide, alumina, magnesium carbonate, tribasic magnesium phosphate, zeolite, hydroxyapatite, and synthetic resin abrasive. The blending amount of the abrasive is adjusted depending on the dosage form, and is preferably 2 to 40%, particularly 10 to 30% for toothpaste, and 0% for liquid toothpaste.

Examples of the thickener include sugar alcohols such as sorbit, xylit, malt, and lactite, and polyhydric alcohols such as glycerin, propylene glycol, and polyethylene glycol. The blending amount is usually 5 to 50%, particularly 20 to 45%. preferable.

As the binder, cellulose derivatives such as sodium carboxymethyl cellulose, methyl cellulose, hydroxyethyl cellulose, gums such as xanthan gum and gum arabic, organic binders such as carrageenan, polyvinyl alcohol, sodium polyacrylate, gelling silica, gel Inorganic binders such as curable aluminum silica, bee gum, and laponite. The blending amount is usually 0.1 to 5% for toothpaste and 0 to 5% for liquid toothpaste and mouthwash.

As the surfactant, an anionic surfactant, a nonionic surfactant, a cationic surfactant, and an amphoteric surfactant can be blended.
Examples of the anionic surfactant include alkyl sulfates such as sodium lauryl sulfate, N-acyl sarcosine salts such as N-lauroyl sarcosine sodium and N-myristoyl sarcosine sodium, N-acyl glutamates such as sodium N-palmitoyl glutamate, Examples include sodium N-methyl-N-acyl taurine, sodium N-methyl-N-acylalanine, sodium α-olefin sulfonate, and the like.
Nonionic surfactants include sugar fatty acid 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, hexaglyceryl monolaurate, Polyglycerin fatty acid esters such as hexaglyceryl monomyristate, decaglyceryl monolaurate, decaglyceryl monomyristic acid, polyoxyethylene sorbitan fatty acid esters such as polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monostearate, polyoxyethylene Polyoxyethylene fatty acid esters such as hydrogenated castor oil and polyoxyethylene higher alcohols such as polyoxyethylene lauryl ether Ethers, fatty acid alkanolamides, such as lauric acid diethanolamide, polyoxyethylene polyoxypropylene copolymers, polyoxyethylene polyoxypropylene fatty acid esters.
Examples of the cationic surfactant include alkyl ammonium and alkyl benzyl ammonium salts, and examples of the amphoteric surfactant include betaines such as alkyl betaines, fatty acid amidopropyl betaines, and alkyl imidazolinium betaines.
The blending amount of the surfactant is preferably 0.001 to 10%, particularly preferably 0.1 to 5% of the entire composition.

Perfumes include peppermint oil, spearmint oil, anise oil, eucalyptus oil, winter green oil, cassia oil, clove oil, thyme oil, sage oil, lemon oil, orange oil, peppermint oil, cardamom oil, coriander oil, mandarin oil, Lime oil, lavender oil, rosemary oil, laurel oil, camomil oil, caraway oil, marjoram oil, bay oil, lemongrass oil, origanum oil, pine needle oil, neroli oil, rose oil, jasmine oil, grapefruit oil, sweetie Natural fragrances such as oil, coconut oil, Iris concrete, absolute peppermint, absolute rose, orange flower, and processing of these natural fragrances (front reservoir cut, rear reservoir cut, fractional distillation, liquid-liquid extraction, essence, Powdered fragrance etc.) and menthol Carvone, Anethole, Cineol, Methyl salicylate, Synamic aldehyde, Eugenol, 3-l-Mentoxypropane-1,2-diol, Thymol, Linalol, Linarel acetate, Limonene, Menthone, Menthyl acetate, N-Substituted-paramenthane 3-carboxamide, pinene, octylaldehyde, citral, pulegone, carbyl acetate, anisaldehyde, ethyl acetate, ethyl butyrate, allylcyclohexane propionate, methyl anthranilate, ethyl methyl phenyl glycidate, vanillin, undecalactone, Hexanal, butanol, isoamyl alcohol, hexenol, dimethyl sulfide, cycloten, furfural, trimethylpyrazine, ethyl lactate, ethyl Oral compositions such as single flavors such as thioacetate, and other flavors such as strawberry flavor, apple flavor, banana flavor, pineapple flavor, grape flavor, mango flavor, butter flavor, milk flavor, fruit mix flavor, tropical fruit flavor, etc. It can be used in combination with known perfume materials used in the above.
Further, the blending amount is not particularly limited, but the above fragrance material is preferably used at 0.000001 to 1% in the preparation composition. Further, as the flavoring fragrance using the fragrance material, it is preferable to use 0.1 to 2% in the preparation composition.

Examples of the sweetener include saccharin sodium, stevioside, paramethoxycinnamic aldehyde, perilartin and the like. Examples of the colorant include blue No. 1, yellow No. 4, titanium dioxide and the like.
Examples of the preservative include benzoic acid such as paraoxybenzoic acid ester and sodium benzoate, or a salt thereof.

As active ingredients, ascorbic acid esters and salts thereof, and those other than methylene blue, for example, nonionic fungicides such as isopropylmethylphenol, cationic fungicides such as cetylpyridinium chloride, chlorhexidine hydrochloride, benzalkonium chloride, benzethonium chloride, Anti-inflammatory agents such as tranexamic acid, epsilon aminocaproic acid, allantoin, glycyrrhetinic acid, glycyrrhizic acid, enzymes such as dextranase, mutanase, amylase, protease, fluorides such as sodium fluoride, sodium monofluorophosphate, orthophosphoric acid Water-soluble phosphate compounds such as potassium salt and sodium salt, copper compounds such as copper gluconate and copper chlorophyllin sodium, sodium chloride, potassium nitrate, aluminum lactate, zinc chloride, zinc citrate, chloride Examples include inorganic salts such as trontium, vitamins such as tocopherol acetate, zeolites, azulene, dihydrocholesterol, chlorophyll, soft extract of thyme, thyme, ogon, clove, hamamelis and other plant extracts, calculus inhibitors, anti-plaque agents, etc. It is done. These active ingredients can be blended in an effective amount as long as the effects of the present invention are not hindered.

Hereinafter, although an Example, a comparative example, and a formulation example are shown and this invention is demonstrated concretely, this invention is not restrict | limited to the following Example. In the following examples, “%” means “% by mass” unless otherwise specified.

[Examples and Comparative Examples]
Sample solutions having the compositions shown in Tables 1 to 3 were prepared by a conventional method, and the cytostatic effect and the bactericidal effect on oral bacteria were evaluated by the following methods. The results are shown in the table.

<Experimental Example 1> Cytotoxicity inhibitory effect test A commercially available gingival fibroblast Gin-1 (manufactured by DS Pharma Biomedical Co., Ltd.) was used in a 10% fetal bovine serum (FBS) -containing Dulbecco's Modified Eagle Medium (D-MEM). And 37 ° C. and 5% CO ₂ . 400 μL of Gin-1 prepared to 5 × 10 ⁴ cells / mL was seeded in a 48-well plate and further cultured for 24 hours. After removing the culture solution, 400 μL of each sample solution shown in the table was added (diluted 20-fold with D-MEM containing 10% FBS) and treated with drugs for 48 hours.
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 removing the solution, 200 μL of a cell activity reagent (Calcinin AM; manufactured by Invitrogen) was added and incubated at 37 ° C. under 5% CO ₂ for 30 minutes. Thereafter, the fluorescence intensity was measured under conditions of Ex / Em = 485 nm / 538 nm using a plate reader (Fluoroskan Ascent; manufactured by Labsystems).
The result calculated from the following formula (1) was taken as the cytotoxicity inhibition rate. Cytotoxicity inhibition test was performed at n = 10. The average value was calculated, and the cytotoxicity-inhibiting effect was evaluated based on the following criteria.
Cytotoxicity inhibition rate (%) = (fluorescence intensity when treated with hydrogen peroxide after treatment with sample solution) / (fluorescence intensity when treated with no hydrogen peroxide after treatment with sample solution) × 100 (1)

Criteria for evaluation of cytostatic effect;
A: Cytotoxicity inhibition rate is 90% or more. O: Cytotoxicity inhibition rate is 70% or more and less than 90%. Δ: Cytotoxicity inhibition rate is 50% or more and less than 70%. X: Cytotoxicity inhibition rate is less than 50%.

<Experimental example 2> Oral bacteria bactericidal effect test Todd Hewitt broth added with hemin (manufactured by Wako Pure Chemical Industries, Ltd., 500 μg / mL) and menadione (manufactured by Wako Pure Chemical Industries, Ltd., 100 μg / mL) A bacterial suspension was prepared from Porphyromonas gingivalis strain ATCC33277 pre-cultured with Nippon Becton Dickinson Co., Ltd. at 1 × 10 ⁹ cells / mL with physiological saline. The bacterial solution and the sample solution shown in the table were mixed at 1: 1 (volume ratio) and allowed to stand for 3 minutes. Thereafter, the solution was diluted 10-fold with physiological saline four times and smeared on a blood agar plate using a spiral plater. As a control, a mixture of physiological saline and bacterial solution at 1: 1 (volume ratio) instead of the drug was used.
The blood plate was cultured at 37 ° C. under anaerobic conditions (95 vol% nitrogen, 5 vol% carbon dioxide) for 5 days, and the number of colonies formed was counted. This oral bacteria sterilization test was conducted with n = 5. The average value was calculated, the number of colonies in the sample solution relative to the number of colonies in the control solution was determined as the viable cell rate, and the bactericidal effect in the oral cavity was evaluated based on the following criteria.

Evaluation criteria for bactericidal effect in oral cavity;
A: The viable cell rate is less than 10% compared to the control. A: The viable cell rate is 10% or more and less than 40% compared to the control. B: The viable cell rate is 40% or more and less than 80% compared to the control. More than 80% viable bacteria compared to control

Details of the raw materials used are shown below.
(A) Magnesium ascorbyl 2-phosphate (Wako Pure Chemical Industries, Ltd.)
Made by Co., Ltd.)
Ascorbic acid-2-phosphate sodium ester (Wako Pure Chemical Industries, Ltd.)
Ascorbic acid-2-sulfate sodium (Wako Pure Chemical Industries, Ltd.)
Made)
(B) Methylene blue (Wako Pure Chemical Industries, Ltd.)

The following are examples of prescriptions. All of the following examples had excellent cytotoxic effect and oral bactericidal effect.

[Prescription Example 1] Toothpaste (A) Ascorbic acid-2-phosphate magnesium 0.2%
(B) Methylene blue 0.01
Silicic anhydride 20.0
Sodium lauryl sulfate 1.0
Propylene glycol 3.0
Sorbit (100% product) 30.0
Sodium carboxymethylcellulose 1.0
Saccharin sodium 0.1
Fragrance 1.0
Purified water balance
Total 100%

[Prescription Example 2] Toothpaste (A) Sodium ascorbyl 2-phosphate 0.5%
(B) Methylene blue 0.005
Silicic anhydride 20.0
Sodium lauryl sulfate 1.0
Propylene glycol 3.0
Sorbit (100% product) 30.0
Sodium carboxymethylcellulose 1.0
Saccharin sodium 0.1
Fragrance 1.0
Purified water balance
Total 100%

[Prescription Example 3] Toothpaste (A) Ascorbic acid-2-sulfate sodium 0.7%
(B) Methylene blue 0.05
Silicic anhydride 20.0
Sodium lauryl sulfate 1.0
Propylene glycol 3.0
Sorbit (100% product) 30.0
Sodium carboxymethylcellulose 1.0
Saccharin sodium 0.1
Fragrance 1.0
Purified water balance
Total 100%

[Formulation Example 4] Liquid dentifrice (A) Ascorbic acid-2-phosphate magnesium 0.3%
(B) Methylene blue 0.001
Xylitol 3.0
Glycerin (100% product) 5.0
Polyoxyethylene (60) hydrogenated castor oil 0.5
Sodium citrate 0.3
Propylene glycol 3.0
Fragrance 0.2
Ethanol 8.0
Purified water balance
Total 100%

[Formulation Example 5] Liquid Dentifrice (A) Ascorbic acid-2-phosphate magnesium 0.3%
(B) Methylene blue 0.001
Xylitol 3.0
Glycerin (100% product) 2.0
Polyoxyethylene (60) hydrogenated castor oil 0.5
Sodium citrate 0.3
Sodium benzoate 0.3
Methyl benzoate 0.1
Propylene glycol 3.0
Saccharin sodium 0.002
Polyethylene glycol (400) 5.0
Fragrance 0.2
Purified water balance
Total 100%

Claims

(A) Ascorbic acid ester or a salt thereof, and (B) methylene blue.
The composition for oral cavity according to claim 1, wherein the component (A) is at least one selected from ascorbic acid phosphate, ascorbic acid sulfate and salts thereof.
The composition for oral cavity according to claim 2, wherein the component (A) is magnesium ascorbate phosphate and / or sodium ascorbate phosphate.
The oral composition according to claim 1, 2 or 3, comprising 0.1 to 2% by mass of component (A) and 0.001 to 0.2% by mass of component (B).
The composition for oral cavity according to any one of claims 1 to 4, wherein (B) / (A) is from 1/2000 to 1/2 as a mass ratio.
The composition for oral cavity according to any one of claims 1 to 5, which is a dentifrice or a mouthwash.