JP4919060B2 - Oral composition and algin dipine inhibitor - Google Patents

Description

The present invention relates to an oral composition and an algindipine inhibitor that are excellent in periodontitis and halitosis suppression effects and are effective in preventing or treating periodontal disease, and further in preventing or suppressing halitosis.

  Periodontal disease is a chronic infectious disease that occurs due to the loss of the balance between the pathogenic action of periodontal disease-causing bacteria and the defense mechanism of the body against the causative bacteria. When inflammation progresses due to infection with the causative bacteria, the gums swell, causing bleeding and pus. Eventually, the alveolar bone that supports the teeth is destroyed, leading to tooth movement and loss. Tooth swaying and loss due to periodontal disease lowers the chewing function and lowers the quality of life of eating things deliciously. Furthermore, it is known that periodontal disease is a risk factor for systemic diseases such as diabetes, heart disease, and childbirth of low-weight infants in pregnant women. Therefore, prevention or treatment of periodontal disease is important not only from the standpoint of maintaining healthy oral function and quality of life, but also from the viewpoint of reducing the risk of systemic diseases.

  For prevention or treatment of periodontal disease, bactericides, antibiotics and anti-inflammatory agents that suppress inflammation are used for the purpose of eliminating the causative bacteria of periodontal disease. However, disinfectants and antibiotics are removed even from resident bacteria other than the causative bacteria, and there are concerns about the risk of causing the emergence of resistant bacteria and bacterial substitution. Further, the anti-inflammatory agent is a coping therapy action that suppresses the inflammation that has occurred, and the cause itself cannot be excluded, and there is a drawback that the possibility of recurrence is high. Therefore, there is a demand for the development of a more effective and safer prevention or treatment technique for periodontal disease that replaces the technique using a fungicide, an antibiotic, or an anti-inflammatory agent.

  A technique using an antibody is a typical one. That is, the antibody is a technology that can prevent or treat periodontal disease without affecting other oral bacteria by specifically binding to and detoxifying pathogenic bacteria and pathogenic factors. It is an effective means that can eliminate itself. However, it is difficult to produce an antibody that does not have a harmful effect on the living body, which has a problem in safety, and has not yet been put into practical use.

  Under such circumstances, algindipine, a protease produced by periodontal pathogens, inhibits neutrophil function, antibody degradation, and periodontal disease, which are important in the defense function against periodontal pathogens. It has been reported that it has an action of degrading extracellular matrix such as collagen constituting the tissue, and it has been elucidated that algindipine is an important pathogenic factor of periodontal disease. Thus, development of an algindipine inhibitor is expected as a technique for preventing or treating periodontal disease specific to a pathogenic factor instead of an antibody.

  Known cysteine protease inhibitors such as antipain, leupeptin, TLCK or E-64 are known as substances that inhibit algindipine, but all of them are highly toxic and used for the prevention and treatment of periodontal disease It has not been. In addition, peptide derivatives containing arginine have been proposed as antagonistic inhibitors that focus on substrate properties that allow arginine pine to selectively degrade the arginine terminus (see Patent Documents 1 and 2). It has not been put into practical use.

  In addition, it has been reported that highly safe protamine, which is generally used for food as a preservative, has an algindipine inhibitory effect (see Non-Patent Document 1), and inhibits algindipine instead of conventional bactericides and antibiotics. (See Patent Documents 3 and 4), but the effect is not sufficient.

  On the other hand, bad breath is a cause of hindering smooth communication, and further, there is a great need for prevention of bad breath from the feeling of not wanting others to feel bad. In general, most of the people who have objective breath odor have causes in the oral cavity, periodontal diseases such as alveolar pyorrhea and dental caries, oral contamination, poor self-cleaning of the oral cavity due to unclean dentures, and This is due to an increase in bad breath-causing bacteria such as Fuzobacterium. When an inflammatory reaction occurs in a tooth or periodontal tissue, the deamination reaction occurs in sulfur-containing amino acids such as blood cell components and epithelial cells, and odorous substances such as hydrogen sulfide, methyl mercaptan, and dimethyl sulfide are generated as volatile sulfides. In addition, if the plaque is not controlled, a volatile sulfide substance decomposed by bacteria in the oral cavity is generated, resulting in bad breath.

  As a bad breath prevention technique, antibacterial action against bad breath causing bacteria and decomposition action of bad breath causing substances by copper or zinc are disclosed (see Patent Document 5). It has also been reported that zinc has an algin dipine inhibitory effect, and that zinc exhibits an algin dipine inhibitory effect synergistic with chlorhexidine and benzamidine (see Non-Patent Documents 2 and 3), and prevention and treatment of periodontal disease. It is also promising as a technology. However, it cannot be said that these technologies also exert a sufficient effect of preventing bad breath, and development of a more effective technology for preventing bad breath is desired.

JP 11-228526 A JP 2001-89436 A Japanese Patent Laid-Open No. 02-250815 Japanese Patent Laid-Open No. 03-261716 Japanese Patent Laid-Open No. 07-165544 Kotani M and 5 others “Inhibitory effects of proteomics on proteolytic and adhesive activities of Porphyromonagingivalis.”, 1999, Infection Immunity, p. 4917-4920 Krauser JA, et al., “Inhibition of argonginingpains (RgpB and HRgpA) with benzamindine inhibitor, vol.8, zinc increases inhibitory, h. 1193-1198 Cronan CA, 3 others, "Inhibition of Porphyromonas gingivalis proteinases (gingipains) by chlorhexidine: synergistic effect of Zn (II)." Oral Microbiol. 212-217

The present invention was made in view of the above circumstances, has a high algindipine inhibitory effect produced by periodontal disease-causing bacteria that are pathogenic factors, is superior in periodontitis suppression effect and halitosis suppression effect, prevention of periodontal disease or An object of the present invention is to provide an oral composition and an algindipine inhibitor effective for treatment, prevention or suppression of halitosis.

  As a result of intensive studies to achieve the above object, the present inventor has (A) protamine and (B) one kind selected from copper gluconate, cupric sulfate, cupric chloride, and cupric acetate. By using together with the above copper compound, a high algindipine inhibitory effect is demonstrated, it is excellent in periodontitis and halitosis suppression effect, and it is effective in preventing or treating periodontal disease, and also in preventing or suppressing halitosis And the present invention has been made.

  In this case, as described above, protamine is generally used as a preservative in foods and has high safety. However, as can be seen from the experimental results described later, it is difficult to say that the effect of inhibiting algindipine is sufficient. In contrast, by blending protamine with a combination of copper compounds selected from the organic and inorganic acid salts of copper described above, these components act synergistically and exhibit a significantly superior argindipine inhibitory effect. In addition, a high gingivitis suppressing effect is exhibited, and a high halitosis suppressing effect is also exhibited. Such a synergistic effect can not be achieved by using protamine in combination with copper chlorophyllin sodium coordinated with copper or a zinc compound known to have an algindipine inhibitory effect, such as zinc gluconate, It is exhibited by the combined use of protamine and the above-mentioned specific copper organic acid salt or inorganic acid salt.

Accordingly, the present invention provides (A) 0.001 to 5% by mass of protamine and (B) one or more copper compounds selected from copper gluconate, cupric sulfate, cupric chloride and cupric acetate. the Ri Na contained and 0.001 to 5 wt%, the oral composition, wherein 0.02 to 50 der Rukoto as mass ratio of component (a) / (B) component and the ( An algin dipine inhibitor comprising an A) component and a (B) component, wherein the (A) component / (B) component is 0.02 to 50 in terms of mass ratio .

  The oral composition of the present invention effectively suppresses inflammation caused by periodontal pathogens, has a high halitosis suppression effect, is highly safe to the human body, and is effective in preventing or treating periodontal disease and preventing or suppressing halitosis It is.

  Hereinafter, the present invention will be described in more detail. The composition for oral cavity of the present invention is selected from (A) protamine and (B) copper gluconate, cupric sulfate, cupric chloride, cupric acetate. 1 or more types of copper compounds are contained.

  The component (A) protamine used in the present invention is a basic protein having a high arginine content that is present in a sperm nucleus such as salmon, trout, herring, mackerel, etc. in association with deoxyribonucleic acid. Extracted and used in the form of sulfate, hydrochloride, etc.

  As protamine, commercially available products generally used as food additives can be used. For example, protamine manufactured by Nichiro Co., Ltd., protamine sulfate manufactured by Organic Synthetic Chemicals Co., Ltd., and the like can be used.

The amount of protamine is 0.001 to 5% of the total composition (mass percentage, hereinafter the same), especially 0.01% to 1% is preferred. The amount is not sufficiently exhibited compounding effect is less than 0.001%, that Do is difficult to stabilize the formulation exceeds 5%.

  Next, the copper compound of the component (B) used in the present invention is one or more copper inorganic acid salts selected from copper gluconate, cupric sulfate, cupric chloride, and cupric acetate. Or it is an organic acid salt. As these copper compounds, not only anhydrides but also hydrates containing crystal water such as cupric sulfate pentahydrate can be used. In the composition of the present invention, one or more of these copper compounds may be contained, but among them, copper gluconate and / or cupric sulfate is particularly preferably used from the point of use (safety) of foods and pharmaceuticals. It is done.

The amount of the copper compound of the component (B) is 0.001 to 5% of the total composition, particularly preferably 0.01% to 1%. The amount is not sufficiently exhibited compounding effect is less than 0.001%, when more than 5% it becomes difficult to stabilize the formulation and problems such as metallic taste is Ru pronounced at.

Furthermore, in the present invention, (A) / (B) is a mass ratio of 0.02 to 50 , preferably 0.1 to 10, in the blending ratio of (A) component protamine and (B) component copper compound. Yes, within the range of the above blending amount so as to be within this range. (A) / or a mass ratio less than 0.01 (B), when it exceeds 100, synergistic effects have such be exhibited satisfactorily by the combination of both components.

  The composition for oral cavity of the present invention essentially contains protamine and a specific copper compound, and is a dentifrice such as a toothpaste, a toothpaste, a liquid dentifrice, a mouthwash, a gel, an ointment, a mouth-cooling agent. It can be prepared in various dosage forms such as pills, tablets for mouthwash, pasta for oral cavity, and gum, and is particularly suitable as a toothpaste, gel, and mouthwash. Here, when preparing into various dosage forms, in addition to the essential components described above, other components can be blended according to the dosage form within a range that does not impair the effects of the present invention, and can be prepared by ordinary methods.

  For example, in the case of dentifrices, various abrasives, wetting agents, binders, surfactants, sweeteners, fragrances, coloring agents, preservatives, other active ingredients, etc. are within a range that does not interfere with the effects of the present invention. Can be used in normal amounts.

  As abrasives, precipitated silica, silica gel, aluminosilicate, zeolite, zirconosilicate, dicalcium phosphate dihydrate and anhydride, calcium pyrophosphate, calcium carbonate, aluminum hydroxide, alumina, magnesium carbonate, tertiary phosphorus Examples thereof include magnesium acid, insoluble sodium metaphosphate, insoluble potassium metaphosphate, titanium oxide, hydroxyapatite, and synthetic resin-based abrasive (mixing amount: usually 5 to 50% based on the entire composition).

  Examples of the wetting agent include glycerin, sorbitol, propylene glycol, polyethylene glycol, and 1,3-butylene glycol (blending amount: usually 10 to 50% based on the entire composition).

  As binder, carrageenan, hydroxyethylcellulose sodium, carboxymethylcellulose sodium, hydroxyethylcellulose, hydroxypropylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, propylene glycol ester alginate, polyacrylic acid, sodium polyacrylate, xanthan gum, tara gum, Guam gum, locust bean gum, gellan gum, gelatin, curdlan, gum arabic, agar, pectin, polyvinyl alcohol, polyvinyl pyrrolidone, pullulan, etc. (blending amount: usually 0.1 to 5% based on the whole composition) .

  As the surfactant, an anionic surfactant, a nonionic surfactant, a cationic surfactant, and an amphoteric surfactant can be blended. Anionic surfactants include sodium alkyl sulfates such as sodium lauryl sulfate and sodium myristyl sulfate, sodium N-acyl sarcosine such as sodium N-lauroyl sarcosine, sodium N-myristoyl sarcosine, sodium dodecylbenzenesulfonate, hydrogenated coconut fatty acid N-acyl glutamate such as sodium monoglyceride monosulfate, sodium lauryl sulfoacetate, sodium N-palmitoyl glutamate, sodium N-methyl-N-acyl taurate, sodium N-methyl-N-acylalanine, sodium α-olefin sulfonate Is mentioned. Nonionic surfactants include sugar alcohol fatty acid esters such as sorbitan fatty acid ester, polyoxyethylene sorbitan fatty acid ester, sucrose fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, polyoxyethylene glycerin fatty acid ester, polyethylene glycol Polyhydric alcohol fatty acid ester such as fatty acid ester, polyoxyethylene alkyl ether, polyoxyethylene polyoxypropylene copolymer, polyoxyethylene alkylphenyl ether, ether type surfactant such as polyoxyethylene hydrogenated castor oil, lauric acid And fatty acid alkanolamides such as diethanolamide. Examples of the cationic surfactant include alkyl ammonium and alkyl benzyl ammonium salts. Examples of amphoteric surfactants include betaine acetate, imidazolinium betaine, and lecithin (blending amount: usually 0.5 to 5% based on the entire composition).

  In addition to the above protamine and water-soluble copper compound, the active ingredient includes, for example, antiplasmin agents such as tranexamic acid and epsilon aminocaproic acid, anti-inflammatory agents such as glycyrrhizin, glycyrrhizinate and allantoins, ascorbate and tocopherol esters Vitamins such as dextranase, mutanase, lysozyme, etc., herbal extracts such as buckwheat extract, hornon extract, clove extract, cetylpyridinium chloride, benzethonium chloride, benzalkonium chloride, decalinium chloride, chlorhexidine hydrochloride, chlorhexidine gluconate , Fungicides such as triclosan, isopropylmethylphenol, hinokitiol, sodium chloride, sodium polyphosphate, sodium metaphosphate, sodium orthophosphate, sodium tripolyphosphate Beam, aluminum lactate, inorganic salts and organic salts such as chitosan and the like. In addition, the compounding quantity of these active ingredients can be made into an effective quantity in the range which does not prevent the effect of this invention.

  Examples of the sweetening agent include saccharin sodium, stevioside, stevia extract, paramethoxycinnamic aldehyde, neohesperidyl hydrochalcone, and perilartin. Examples of the colorant include blue No. 1, yellow No. 4, titanium dioxide and the like. Examples of the preservative include paraoxybenzoic acid ester and sodium benzoate.

  Examples of the fragrances include terpenes such as l-menthol, carvone, anethole and limonene, or derivatives thereof, peppermint oil, and the like.

  Examples of the pH adjuster include citric acid, fumaric acid, malic acid, tartaric acid, and lactic acid.

EXAMPLES Hereinafter, although an experimental example, an Example, and a comparative example are shown and this invention is demonstrated concretely, this invention is not restrict | limited to the following Example. In the following examples, the blending amount is mass%.
In the following examples, protamine is manufactured by Nichiro Co., Ltd., copper gluconate is manufactured by Tomita Pharmaceutical Co., Ltd., cupric sulfate, cupric chloride, cupric acetate, and copper chlorophyllin sodium are all manufactured by Shonan Wako Co., Ltd. A thing was used.

[Experimental Example 1] Examination of Alzin Dipain Inhibitory Activity Effect The lower jaw 4 sites (P3, P4, M1 mesial, M2 centrifugation) where gingivitis is observed in beagle dogs (5 years old, female) were used as test sites. 0.5 mL of the gel preparation having the composition shown in FIG. 3 was applied to the test site three times every 3 hours with a syringe. Gingival crevicular fluid (GCF) before treatment and 3 hours after final treatment was collected by inserting GCF collection strips (Periopaper (registered trademark), manufactured by Yoshida Co., Ltd.) into the periodontal pocket of each test site for 30 seconds. . GCF collection strips were immersed in 100 mmol of 10 mmol / L sodium phosphate buffer (pH 6.0) ^{* 1} to extract GCF, and the argindipine activity of the GCF extract was measured ^{* 2} .
From the ratio of the algindipine activity per unit GCF mass before and after the test drug treatment, the inhibitory effect of algindipine in GCF was determined by the following formula. The amount of GCF collected was determined from the difference in mass of GCF collection strips before and after GCF collection.
The results are shown in Table 1.

Argindipine inhibitory effect (%) = {(MN) / M} × 100
M; (Algindipine activity before drug treatment) / (GCF mass collected before drug treatment)
N: (Algindipain activity after drug treatment) / (GCF mass collected after drug treatment)

^{* 1} Sodium phosphate buffer Sodium dihydrogen phosphate dihydrate (Wako Pure Chemical Industries, Ltd.): 0.156g
1N sodium hydroxide (manufactured by Wako Pure Chemical Industries): appropriate amount (adjusted to pH 6.0)
Distilled water: remaining
(Total volume up to 100 mL)

^{* 2} Measurement method of algindipine activity 20 mmol / L sodium phosphate buffer (pH 7.5) containing 5 mmol / L cysteine as a reaction buffer ^{* 3} , GCF extract 10 μL, and substrate (10 μmol / L prepared with reaction buffer) 140 μL of Bz-Arg-MCA: carbobenzoxy-L-phenylalanyl-L-arginine-4-methylcoumaryl-7-amide (manufactured by Peptide Institute) was mixed and reacted at 37 ° C. for 15 minutes. The fluorescence intensity of AMC (7-amino-4-methylcoumarin) produced by the reaction was measured at an excitation wavelength of 390 nm and a measurement wavelength of 460 nm (Fluoroscan Ascent, manufactured by Dainippon Pharmaceutical Co., Ltd.), and defined as the activity of algindipine. That is, the stronger the fluorescence intensity, the higher the algindipine activity.

^{* 3} Reaction buffer: mass in 100 mL L-cysteine (manufactured by Sigma Aldrich): 0.0788 g
Sodium dihydrogen phosphate dihydrate (Wako Pure Chemical Industries, Ltd.): 0.312 g
1N sodium hydroxide (manufactured by Wako Pure Chemical Industries, Ltd.): appropriate amount (adjusted to pH 7.5)
Distilled water: remaining
(Total volume up to 100 mL)

  As is clear from Table 1, it was found that the combined use of protamine and copper gluconate, cupric sulfate, cupric chloride or cupric acetate synergistically improves the arginine dipine inhibitory effect. This improving effect is not observed when copper chlorophyllin sodium coordinated with copper or zinc gluconate, which is a zinc compound known to have an inhibitory effect on algindipain, is used in combination with protaamine. It was also found that this action is specific to copper compounds composed of organic acid salts.

[Experimental Example 2] Examination of improvement effect of gingivitis and bad breath The improvement effect of gingivitis and bad breath by algin dipain inhibitory activity was verified by the following method.
For adult males, perform gingivitis assessment ^{* 4} and halitosis assessment ^{* 5} , with a gingivitis score of 0.5 to less than 1.5 and a bad breath intensity of 2 to less than 3 Were selected as subjects. The test subjects were divided into 7 groups (5 in each group) so that the average scores of gingivitis and bad breath were almost the same value, and included about 15 mL of mouthwash with the composition shown in Table 2 twice a day. After rinsing for about 20 seconds, it was brushed with a toothbrush. After 4 weeks, gingivitis and halitosis were evaluated again, and the gingivitis suppressing effect and halitosis suppressing effect of the test composition were determined according to the following evaluation criteria. The results are shown in Table 2.

^{* 4} Evaluation of gingivitis For 6 representative teeth (left and right upper jaws 2, 6 and lower jaw 4), the lingual side of each tooth, the mesial side of the lingual side, the distal side and the buccal side of the tooth, Gingivitis was evaluated according to the following evaluation criteria for a total of 24 sites (6 teeth × 4 sites), and the average value of 24 sites was determined. Furthermore, the gingivitis improvement degree was calculated | required by the following formula, and the gingivitis suppression effect was evaluated by the following evaluation criteria.

<Evaluation criteria for gingivitis>
0 point: No inflammation is observed.
1 point: Mild inflammation. There is a slight color change, but no bleeding is observed by probing.
2 points: moderate inflammation. There is redness, edema, swelling, and bleeding is observed by probing.
3 points: Severe inflammation. Significant redness and spontaneous bleeding are observed.

<Calculation formula for improvement of gingivitis>
(Gingivitis improvement) = (A−B) / (Number of subjects in each group)
A: (Total of gingivitis score average of 24 sites before test)
B: (Total of gingivitis scores after the test, average of 24 sites)

<Evaluation criteria for gingivitis inhibitory effect of test composition>
A: There is a remarkable effect. Gingivitis improvement is 0.5 points or more ○: Effective. Gingivitis improvement degree is 0.25 point or more and less than 0.5 point Δ: Some effect. Gingivitis improvement is greater than 0 and less than 0.25 ×: No effect. Gingivitis improvement is 0 point or less

^{* 5} Evaluation of bad breath The expert evaluator evaluated the bad breath strength of 1 liter of breath collected in a Tedora bag according to the following evaluation criteria. Furthermore, the bad breath improvement degree was calculated | required by the following formula, and the bad breath suppression effect of the test composition was calculated | required by the following evaluation criteria.

<Evaluation criteria for bad breath intensity>
0 point: Bad breath is not recognized.
1 point: A slight bad breath is recognized.
2 points: Bad breath is observed.
3 points: Significant bad breath is observed.

<Calculation formula of bad breath improvement>
(Degree of bad breath improvement) = (Average value of bad breath before test start)-(Average bad breath intensity after test)
In addition, a bad breath intensity average value is the value which averaged the bad breath intensity of five test subjects in each group.

<Evaluation criteria for the bad breath suppression effect of the test composition>
A: There is a remarkable effect. Bad breath improvement degree is 1.5 points or more. Bad breath improvement degree is 0.5 point or more and less than 1.5 point Δ: Some effect. Bad breath improvement degree is 0.2 point or more and less than 0.5 point ×: No effect. Bad breath improvement is less than 0.2 points.

  As is apparent from Table 2, the combined use of protamine and copper gluconate provides a high gingivitis suppression effect and bad breath suppression effect, and is particularly effective when protamine / copper gluconate is 0.1 to 10 by mass ratio. Was confirmed to be high.

[Example 10] Ointment protamine 0.05
Cupric sulfate 0.5
Glyceryl monomyristate 5
Citric acid 0.1
Liquid paraffin 15
White petrolatum balance
Total 100.0%

[Example 11] Protamine gel agent 0.01
Copper gluconate 0.5
Hydroxyethylcellulose 1.5
Polyethylene glycol 4000 15
Ethyl paraoxybenzoate 0.03
Water balance
Total 100.0%

[Example 12] Dentifrice (Toothpaste)
Calcium phosphate 25.0
Silicic anhydride 5.0
Sodium lauryl sulfate 1.2
Carboxymethylcellulose 0.8
Carrageenan 0.5
Sodium fluoride 0.2
Protamine 0.01
Copper gluconate 0.05
Sorbitol 15.0
Fragrance 1.0
Water balance
Total 100.0%

[Example 13] Dentifrice (Toothpaste)
Silicic anhydride 15.0
Sodium lauryl sulfate 1.0
Xanthan gum 0.5
Sodium alginate 0.5
Saccharin sodium 0.1
Sodium monofluorophosphate 0.1
Protamine 0.02
Copper gluconate 0.05
Sorbitol 10.0
Xylitol 5.0
Fragrance 1.0
Water balance
Total 100.0%

[Example 14] Mouthwash ethanol 10.0
Protamine 0.01
Copper gluconate 0.05
Tranexamic acid 0.05
Sorbitol 5.0
Xylitol 5.0
Polyoxyethylene (60) hydrogenated castor oil 0.5
Saccharin sodium 0.2
Fragrance 0.8
Water balance
Total 100.0%

[Example 15] Gum gum base 20.0
Fragrance 1.0
Minamata 20.0
Powdered sugar 10.0
Trisodium citrate 1.5
Protamine 0.01
Copper gluconate 0.3
Sorbitol 15.0
Xylitol 10.0
Water balance
Total 100.0%

Claims (2)

(A) 0.001-5 mass% of protamine , and (B) 1 or more types of copper compounds chosen from copper gluconate, cupric sulfate, cupric chloride, cupric acetate 0.001-5. Ri Na contained mass%, (a) component / (B) oral composition in which the components and wherein 0.02 to 50 der Rukoto as a mass ratio. It consists of (A) protamine and (B) one or more copper compounds selected from copper gluconate, cupric sulfate, cupric chloride and cupric acetate, and (A) component / (B) component An algin dipine inhibitor having a mass ratio of 0.02 to 50.