JP2005132768A - Composition for oral cavity - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a safe composition for oral cavity effective for preventing and treating periodontoses. <P>SOLUTION: The composition for oral cavity contains one or more compounds selected from dehydroabietic acid, hydroxyabietic acid, tetrahydroxyabietic acid and their salts. More specifically, this composition is to be used for preventing/treating periodontoses by effectively inhibiting matrix metalloprotease activity by the above compound(s). <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

The present invention relates to an oral composition containing one or more selected from dehydroabietic acid, hydroxyabietic acid, tetrahydroxyabietic acid and salts thereof. More specifically, one or more selected from dehydroabietic acid, hydroxyabietic acid, tetrahydroxyabietic acid and salts thereof effectively prevent matrix metalloprotease activity, thereby preventing or treating periodontal disease. The present invention relates to an oral composition.

  Matrix metalloprotease (MMP) is a general term for metalloenzymes that degrade extracellular matrix. The extracellular matrix is a biopolymer that fills gaps between cells in mammalian tissue, and contains collagen, proteoglycan, elastin, and the like as components. The metabolism of this extracellular matrix is mainly regulated by the balance between matrix metalloprotease and tissue-derived metalloprotease inhibitor (TIMP) specific for matrix metalloprotease. Abnormal structure of extracellular matrix components and disruption of synthesis / degradation metabolic balance are associated with joint diseases such as osteoarthritis and rheumatoid arthritis, as well as cancer invasion and metastasis. And increased matrix metalloprotease activity are known to be related.

  As matrix metalloproteases, more than 10 types of enzyme molecules such as collagenase (MMP-1 and 8), stromelysin (MMP-3), gelatinase (MMP-2 and 9) are known (Yoshihara, new name: inflammation) And immunization, 2, 177-185, 1994), these are produced by a variety of cells. Collagenase is not normally produced from epithelial cells, but in periodontal tissues it is frequently expressed around the adherent and pocket epithelium (M. Kylmaniemi et al: J. Dent Res., 75: 919-926). 1996). In addition, gelatinase that degrades type IV collagen, which is a component of the basement membrane, has been reported to be produced from epithelial cells by stimulation of factors such as lipopolysaccharide derived from periodontal disease-causing bacteria. Matrix metalloproteinases are thought to be deeply involved in the progression of periodontal disease.

In addition, the amount of matrix metalloprotease in oral irrigation fluid, gingival crevicular fluid and saliva of periodontal disease patients has been reported to reflect the patient's pathology, and the amount of matrix metalloprotease decreases with treatment (M. Makela et al: J. Dent Res., 73: 1397-1406, 1994). It has also been reported that when the amounts of matrix metalloprotease and TIMP are compared with those in healthy subjects, it is considerably higher in patients with gingivitis and higher in patients with periodontitis (A. Haerian et al: J. Clin Periodontol. 22: 505-509, 1995).

To date, tetracycline and its modifications have been reported to inhibit matrix metalloprotease activity in fibroblasts and epithelial cells (L. H. Nip et al: J. Periodont Res., 28: 379-). 385, 1993). In addition, as inhibitors for matrix metalloproteases, a number of hydroxamic acid derivatives (JP 7-505387 A, JP 08-81443 A), esculetin derivatives (JP 08-183785 A) and the like are synthesized, A flavone derived from a natural product or an anthocyanidin as an active ingredient has also been reported (Japanese Patent Laid-Open No. 08-104628). In addition, it has been reported that quinones derived from natural products, tricyclic diterpenes and pentacyclic triterpenes have an inhibitory effect on matrix metalloprotease activity and / or an inhibitory effect on cellular matrix metalloprotease production. (JP-A-11-139947)

Yoshiwara, Shinna: Inflammation and immunity, 2, 177-185, 1994 M. Kylmaniemi et al: Dent Res. 75: 919-926, 1996 M ... Makela et al: J.A. Dent Res. , 73: 1397-1406, 1994 A. Haerian et al: J.A. Clin Periodontol. , 22: 505-509, 1995 L. H. Nip et al. Periodont Res. , 28: 379-385, 1993 JP 07-505387 JP 08-81443 A Japanese Patent Laid-Open No. 08-183785 Japanese Patent Laid-Open No. 08-104628 JP-A-11-139947

  An object of the present invention is to provide a composition for oral cavity that inhibits matrix metalloprotease activity and is excellent in preventing and treating periodontal disease.

In order to achieve the above object, the present inventors have conducted intensive research, and as a result, dehydroabietic acid, hydroxyabietic acid, tetrahydroxyabietic acid and salts thereof have an inhibitory effect on the activity of matrix metalloprotease. The present inventors have found that an oral composition containing this composition is effective for the prevention and treatment of periodontal disease, and has completed the present invention.

That is, this invention provides the composition for oral cavity which inhibits the activity of a matrix metalloprotease. ADVANTAGE OF THE INVENTION According to this invention, the composition for oral cavity which is safe and effective in prevention and treatment of periodontal disease can be provided.

The present invention
Item 1 An oral composition comprising one or more selected from dehydroabietic acid, hydroxyabietic acid, tetrahydroxyabietic acid, and salts thereof.
Item 2. The oral composition according to Item 1, wherein the oral composition is a mouthwash.
Item 3. The oral composition according to any one of Items 1 and 2, wherein the oral composition is used for preventing periodontal disease.

The present invention is described in detail below.
Components that inhibit the activity of the matrix metalloprotease used in the present invention are abietic acid derivatives such as dehydroabietic acid, hydroxyabietic acid, and tetrahydroxyabietic acid, and salts thereof, and the like, and dehydroabietic acid is particularly preferable. Dehydroabietic acid is a component obtained by purifying a resin contained in pine, and is commercially available, for example, as KE-604 (manufactured by Arakawa Chemical Co., Ltd.).
The amount of the component that inhibits the activity of the matrix metalloprotease such as dehydroabietic acid of the present invention is 0.001 to 10% by weight, and particularly preferably 0.01 to 3.0% by weight. When the blending amount is less than 0.001% by weight, a sufficient effect cannot be obtained, and when the blending amount exceeds 10% by weight, it becomes difficult to formulate as an oral composition.

In the present invention, the activity inhibiting effect of matrix metalloproteinase means that matrix metalloprotease produced by periodontal tissue cells affected by periodontal disease etc. degrades extracellular matrix between tissues and worsens tissue destruction. Is to inhibit. The production inhibitory effect of matrix metalloprotease is effective for the prevention and treatment of symptoms such as periodontitis, gingivitis, periodontitis, periodontitis, periodontal disease such as peri-implantitis, and the like.
Furthermore, in addition to the components of the present invention, it is expected that various symptoms of periodontal disease or the prevention and treatment effect of the disease will be improved when a conventionally known disinfectant is added. Specifically, quaternary ammonium salts, bisbiguanides, and phenols may be considered, but quaternary ammonium salts and bisbiguanides that are cationic fungicides are preferred, and cetylpyridinium chloride is particularly preferred. preferable. The blending amount of these bactericides is 0.001 to 0.5% by weight, particularly 0.01 to 0.1% by weight.

The oral composition of the present invention can be provided as a toothpaste, a liquid toothpaste, a mouthwash, a mouth gel, etc., particularly in the optimal form of a mouthwash, depending on its form. is there. In the form of these oral compositions, it can be produced by a normal production method. In particular, in the case of a mouthwash, a method of producing a porous material under high pressure is also effective.
In the present invention, in addition to the essential components described above, other components such as abrasives, wetting agents, monohydric alcohols, binders, fragrances, sweeteners, pH adjusters, preservatives, dyes, etc. It can mix | blend suitably in the range which does not impair the effect of this invention.

As an abrasive, calcium carbonate, calcium phosphate, dicalcium phosphate, calcium pyrophosphate, insoluble sodium metaphosphate, titanium oxide, amorphous silica, crystalline silica, aluminosilicate, aluminum oxide, aluminum hydroxide, resin, etc. alone or 2 It can mix | blend combining a seed | species or more. These compounding quantities are 10-60 weight% normally with respect to the composition whole quantity.

As the wetting agent, for example, polyhydric alcohols such as glycerin, sorbitol, polyethylene glycol, propylene glycol, ethylene glycol, 1,3-butylene glycol, polypropylene glycol, xylitol, maltitol, lactitol, alone or in combination of two or more. The blending amount is usually 3 to 20% by weight based on the entire composition.

Examples of the monohydric alcohol include ethanol, propyl alcohol, and isopropyl alcohol, and ethanol is particularly preferable. These monohydric alcohols can be blended singly or in combination of two or more, and the blending amount is usually 0 to 15% by weight with respect to the whole composition.

Examples of the binder include cellulose derivatives such as carrageenan and carboxymethyl cellulose, alkali metal alginates such as sodium alginate, gums such as xanthan gum, tragacanth gum and gum arabic, and synthetic binders such as polyvinyl alcohol and sodium polyacrylate. Examples thereof include inorganic binders such as silica gel, aluminum silica gel, and bee gum. The amount of these binders is usually 0.01 to 5% by weight based on the total amount of the composition.

Examples of flavoring agents include anethole, menthol, peppermint oil, spearmint oil, lemon oil, orange oil, sage oil, rosemary oil, cinnamon oil, perilla oil, winter green oil, clove oil, eucalyptus oil, pimento oil, Carvone, cinnamic aldehyde, cineol, menthone, limonene, methyl salicylate and the like can be blended alone or in combination of two or more thereof within a range not impairing the effects of the present invention. The amount is usually 0.0001 to 1.0% by weight based on the total amount of the composition.

Examples of the sweetening agent include palatinit, saccharin sodium, acesulfame potassium, stevioside, neohesperidyl dihydrochalcone, glycyrrhizin, perilartine, thaumatin, asparatylphenylalanyl methyl ester, ρ-methoxycinnamic aldehyde, and the like. Can be blended alone or in combination of two or more. The compounding quantity is 0.01 to 1 weight% normally with respect to the whole composition, Preferably it is 0.05 to 0.5 weight%.

Examples of the pH adjuster include citric acid, phosphoric acid, malic acid, pyrophosphoric acid, lactic acid, tartaric acid, glycerophosphoric acid, acetic acid, nitric acid, or a chemically possible salt thereof, sodium hydroxide, and the like. Can be blended alone or in combination of two or more so that the pH of the composition is in the range of 5-9. The amount is usually 0.01 to 2% by weight based on the entire composition.

Furthermore, the oral composition of the present invention includes vitamin Es such as dl-α-tocopherol acetate, tocopherol succinate, or tocopherol nicotinate, dextranase, amylase, protease, mutanase, lysozyme, lytic enzyme (lytech enzyme). ), Enzymes such as tranexamic acid and epsilon aminocaproic acid, aluminum chlorohydroxyl allantoin, dihydrocholesterol, glycyrrhizin salts, glycyrrhetinic acid, glycerophosphate, chlorophyll, sodium chloride, caropeptide, water-soluble inorganic phosphate compounds, It can mix | blend individually or in combination of 2 or more types. The composition for oral cavity of this invention can mix these components and can manufacture it according to a normal method.

Hereinafter, the present invention will be described more specifically with reference to test examples and examples. However, the present invention is not limited to the following test examples and examples. Further, unless otherwise specified, [%] is [% by weight].

Test Example 1 Measurement of inhibitory activity against matrix metalloprotease Material 1-1
Test compound: dehydroabietic acid 1-2
Preparation of reaction solution The test compound was dissolved in methanol and diluted 100-fold or more with a substrate buffer (5 mM CaCl ₂ , 50 mM Tris buffer containing 1 mM ZnCl ₂ , pH 7.4). At this time, the test was carried out at 3 concentrations (10 μg / ml, 30 μg / ml, 100 μg / ml) for each compound.

1-3
Cells and culture conditions Gingival fibroblasts (HGF cells) were primary cultured and passaged in Dulbecco's modified Eagle medium (DMEM) containing 10% fetal bovine serum (FBS).

1-4
Cell-treated HGF cells were seeded in a 10 cm diameter dish and cultured in DMEM medium supplemented with 10% serum until confluent, and then the cell surface was washed with serum-free DMEM. After washing, the cells were cultured in serum-free DMEM for 48 hours, the culture supernatant was transferred to a filter, and centrifuged and concentrated. The concentrated solution was used for the experiment.

2. Measurement of inhibitory effect on collagenase activity The inhibitory effect of each test compound on HGF cell-derived collagenase was measured by gelatin zymography using gelatin as a substrate. After separating the enzyme protein by slab SDS-PAGE, collagenase is observed as a colorless band on a blue background (gelatin gel) by reacting with a substrate coexisting in the gel. Slab electrophoresis was performed according to the Laemli method, and gelatin was added to a separation gel of 10% acrylamide to a final concentration of 0.3 mg / ml. Cell supernatant was applied and run.
After electrophoresis, the gel was immersed in Triton buffer (2.5%, Triton X-100-containing 50 mM Tris buffer, pH 7.5), and SDS was removed while shaking for 20 minutes. Next, it was immersed in a substrate buffer containing the test compound and reacted at 37 ° C. for 20 hours. As a negative control, a test compound to which no test compound was added was used. After the enzyme reaction, it was stained with 0.02% Coomassie brilliant blue R250 and decolorized until a band could be observed with an acetic acid / methanol aqueous solution.

  The area of the MMP activity band when the test compound was added was classified according to the following criteria in comparison with the area when no test compound was added (negative control), and the MMP activity inhibitory effect was evaluated. The measurement results of enzyme inhibitory activity are shown in Table 1.

Evaluation criteria ◎ ・ ・ ・ Inhibitory activity: Strong (no band visible at all)
○ ... Inhibitory activity: Medium (band width is about 1/2)
Δ: Inhibitory activity: weak (band width is about 2/3 to 3/4)
X: Inhibitory activity: None (the band width does not change at all)

Furthermore, preparations were prepared according to the following formulas according to a conventional method.
Example 1 Toothpaste
Ingredient amount%
Dehydroabietic acid 1.0
Dl-a-tocopherol acetate 0.05
Chlorhexidine hydrochloride 0.05
Sorbit 40.0
Silicic anhydride 20.0
Sodium carboxymethylcellulose 1.3
Sodium lauryl sulfate 1.0
Methyl paraoxybenzoate 0.1
Saccharin sodium 0.1
Anethole 0.1
Menthol 0.2
Peppermint oil 0.4
Glycerin 5.0
Purified water remaining
Total 100.0

Example 2 Toothpaste
Ingredient amount%
Dehydroabietic acid 0.1
Cetylpyridinium chloride 0.05
Sodium monofluorophosphate 0.70
Calcium hydrogen phosphate 35.0
Sorbit liquid 25.0
Alkyl glucoside 1.0
Hydroxyethyl cellulose 1.0
Paraoxybenzoic acid ester 0.1
Propylene glycol 2.0
Saccharin sodium 0.3
Peppermint 0.2
Methyl salicylate 0.1
Anethole 0.2
Menthol 0.3
Cineol 0.1
Purified water remaining
Total 100.0

Example 3 Mouthwash
Ingredient amount%
Dehydroabietic acid 0.2
Palatinit 30.0
Pluronic 1.0
Citric acid 0.1
Sodium citrate 0.3
Stevia extract 0.2
Anethole 0.2
Peppermint 0.5
Spearmint 0.05
Ethanol 10.0
Sorbitol 10.0
Purified water remaining
Total 100.0

Example 4 Toothpaste
Ingredient amount%
Dicalcium phosphate 30.0
Glycerin 10.0
Sorbitol 20.0
Sodium carboxymethylcellulose 1.0
Sodium lauryl sulfate 1.5
Carrageenan 0.5
Saccharin sodium 0.1
Fragrance 1.0
Sodium benzoate 0.3
Hydroxyabietic acid 0.2
Dipotassium glycyrrhizinate 0.05
Purified water remaining
Total 100.0

Example 5 Mouthwash
Ingredient amount%
Ethanol 10.0
Glycerin 5.0
Citric acid 0.01
Sodium citrate 0.1
Polyoxyethylene (60EO) hydrogenated castor oil 0.5
Methyl paraoxybenzoate 0.1
Fragrance 0.2
Dehydroabietic acid 0.05
Triclosan 0.2
Purified water remaining
Total 100.0

Example 6 Mouthwash
Ingredient amount%
Ethanol 10.0
Tranexamic acid 0.05
Glycerin 5.0
Citric acid 0.01
Sodium citrate 0.1
Polyoxyethylene hydrogenated castor oil 0.5
Methyl paraoxybenzoate 0.1
Tetrahydroxyabietic acid 0.1
Birch extract 0.5
Purified water remaining
Total 100.0

Example 7 Oral Pasta
Ingredient amount%
Liquid paraffin 13.0
Cetanol 10.0
Glycerin 25.0
Sorbitan monopalmitate 0.6
Polyoxyethylene sorbitan monostearate 5.0
Sodium lauryl sulfate 0.1
Benzethonium chloride 0.1
Tocopherol nicotinate 0.2
Cetylbenzethonium chloride 0.05
Methyl salicylate 0.1
Saccharin 0.2
Fragrance 0.25
Rosemary extract 0.2
α-Amylin 0.05
Dehydroabietic acid 0.5
Purified water remaining
Total 100.0

Example 8 Oral Pasta
Ingredient amount%
Carboxymethylcellulose 0.2
Glycerin 40.0
Dehydroabietic acid 1.0
Purified water remaining
Total 100.0

Example 9 Mouthwash
Ingredient amount%
Ethanol 10.0
Glycerin 5.0
Citric acid 0.01
Sodium citrate 0.1
Polyoxyethylene (60EO) hydrogenated castor oil 0.5
Methyl paraoxybenzoate 0.1
Hydroxyabietic acid 0.1
Cetylpyridinium chloride 0.5
Purified water remaining
Total 100.0

Claims (3)

An oral composition comprising one or more selected from dehydroabietic acid, hydroxyabietic acid, tetrahydroxyabietic acid, and salts thereof. The oral composition according to claim 1, wherein the oral composition is a mouthwash. The composition for oral cavity according to any one of claims 1 and 2, wherein the composition for oral cavity is for preventing periodontal disease.