Oral components

Abstract

To inhibit protease activity of gingipain.SOLUTION: An oral composition for inhibiting the protease activity of gingipain is provided, the oral composition containing yeast extract. The oral composition further contains at least one selected from citrate and polysorbate.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an oral composition. [Background technology]

[0002] As disclosed in Patent Document 1, Porphyromonas gingivalis (hereinafter referred to as "gingivalis") is known as a bacterium associated with periodontal disease. Porphyromonas gingivalis produces gingipain, a type of protease. Gingipains include Arg-gingipain (Rgp) and Lys-gingipain (Kgp), which have different peptide cleavage site specificities. It has been found that the protease activity of gingipain causes the onset and progression of periodontal disease. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Patent Publication No. 2021-31399 Summary of the Invention [Problem to be solved by the invention]

[0004] It is believed that inhibiting the protease activity of gingipains will lead to the prevention of periodontal disease and the suppression of its progression. There is a need for an oral composition that can inhibit the protease activity of gingipains. [Means for solving the problem]

[0005] The oral composition for solving the above problem contains yeast extract and further contains at least one selected from citrate salts and polysorbates, and is intended to inhibit the protease activity of gingipain.

[0006] In the oral composition, the citrate is preferably sodium citrate or ammonium citrate. The oral composition preferably contains at least one selected from the citrate salts and at least one selected from the polysorbates. [Effects of the Invention]

[0007] The oral composition of the present invention can inhibit the protease activity of gingipain. DETAILED DESCRIPTION OF THE INVENTION

[0008] One embodiment of the oral composition will be described below. The oral composition of this embodiment contains yeast extract. The oral composition further contains at least one selected from a specific citrate salt and a specific emulsifier. The oral composition is for inhibiting the protease activity of gingipain. Hereinafter, the protease activity of gingipain may also be referred to as gingipain activity.

[0009] <Yeast extract> The raw material for the yeast extract is not particularly limited, and known yeasts can be used. The yeast extract can be one produced by a known production method. The yeast extract is, for example, an extract obtained by autolysis of yeast. As the yeast extract, it is preferable to use, for example, an extract obtained by decomposing the cells of baker's yeast (Saccharomyces cerevisiae).

[0010] The content of yeast extract in the oral composition is not particularly limited, but is preferably 0.1% by mass or more and 1.0% by mass or less. The lower limit of the yeast extract content is more preferably 0.3% by mass or more, and even more preferably 0.5% by mass or more. The upper limit of the yeast extract content is more preferably 0.8% by mass or less, and even more preferably 0.6% by mass or less.

[0011] <Citrate> Examples of citrates include sodium citrate, potassium citrate, calcium citrate, and ammonium citrate. It is particularly preferable for the oral composition to contain sodium citrate or ammonium citrate. One type of citrate may be used alone, or two or more types may be used in combination.

[0012] The content of citrate in the oral composition is not particularly limited, but is preferably 0.1% by mass or more and 0.5% by mass or less. The lower limit of the citrate content is more preferably 0.15% by mass or more, and even more preferably 0.2% by mass or more. The upper limit of the citrate content is more preferably 0.4% by mass or less, and even more preferably 0.3% by mass or less.

[0013] The mass ratio of yeast extract to citrate (yeast extract / citrate) is not particularly limited, but is preferably 1 or more and 10 or less. The lower limit of this mass ratio is more preferably 2 or more, and even more preferably 2.5 or more. The upper limit of this mass ratio is more preferably 5 or less, and even more preferably 3 or less.

[0014] <emulsifier> Examples of emulsifiers include anionic surfactants, amphoteric surfactants, and nonionic surfactants. A nonionic surfactant is preferably used as the emulsifier. Polysorbates are preferred as nonionic surfactants. Examples of polysorbates include polysorbate 80, polysorbate 60, and polysorbate 20. It is particularly preferred that the oral composition contains polysorbate 80. One emulsifier may be used alone, or two or more emulsifiers may be used in combination. For example, polysorbate 80 may be combined with other polysorbates.

[0015] The content of the emulsifier in the oral composition is not particularly limited, but is preferably 0.05% by mass or more and 0.2% by mass or less. The lower limit of the emulsifier content is more preferably 0.08% by mass or more, and even more preferably 0.1% by mass or more. The upper limit of the emulsifier content is more preferably 0.18% by mass or less, and even more preferably 0.15% by mass or less.

[0016] The mass ratio of yeast extract to emulsifier (yeast extract / emulsifier) ​​is not particularly limited, but is preferably from 1 to 20. The lower limit of this mass ratio is more preferably 3 or more, and even more preferably 5 or more. The upper limit of this mass ratio is more preferably 10 or less, and even more preferably 7 or less.

[0017] <Application form, use, and dosage form> The application form of the oral composition is not particularly limited, and can be used as, for example, a pharmaceutical, a quasi-drug, or a cosmetic. Known uses of the oral composition can be appropriately adopted. Examples of uses include chewing agents, oral dissolving agents, oral disintegrating agents, tongue care agents, mouth fresheners, toothpastes, mouthwashes, gargles, liquid dentifrices, biofilm dispersants, anti-halitosis agents, gum massage agents, oral moisturizing agents, tongue coating removers, oral application agents, oral disinfectants, throat disinfectants, oral throat agents, periodontal disease treatment agents, denture adhesives, denture coating agents, denture stabilizers, denture preservatives, denture cleaners, and implant care agents.

[0018] The dosage form of the oral composition is not particularly limited, and for example, by containing a solvent such as water or alcohol, it can be applied to ointments, pastes, sprays, gels, liquids, suspensions, gums, etc.

[0019] The type of water used as a solvent is not particularly limited, and examples thereof include distilled water, pure water, ultrapure water, purified water, and tap water. The type of alcohol used as a solvent is not particularly limited, and examples thereof include ethanol. A mixture of water and alcohol can also be used.

[0020] When the oral composition is in a liquid form, the content of the solvent such as water is not particularly limited, but is preferably 60% by mass or more and 99.8% by mass or less, and more preferably 70% by mass or more and 90% by mass or less.

[0021] <Other ingredients> The oral composition may contain other ingredients in addition to the ingredients described above depending on the intended application, form, use, etc. Examples of other ingredients include antibacterial agents, anti-inflammatory agents, fragrances, humectants, abrasives, alcohols, thickeners, sweeteners, medicinal ingredients, colorants, stabilizers, and pH adjusters. Known ingredients that are incorporated into oral compositions can be used as other ingredients. The oral composition may contain only one of the above other ingredients alone, or may contain two or more of them in combination.

[0022] Examples of antibacterial agents include cetylpyridinium chloride, paraben, sodium benzoate, triclosan, chlorhexidine hydrochloride, isopropylmethylphenol, benzalkonium chloride, benzethonium chloride, and hinokitiol.

[0023] Anti-inflammatory agents include, for example, glycyrrhizinate, tranexamic acid, ε-aminocaproic acid, and Phellodendron Bark extract. Examples of fragrances include anethole, eugenol, carvone, wintergreen, methyl salicylate, thymol, clove oil, sage oil, ocimene oil, and citronellol.

[0024] Examples of abrasives include calcium carbonate, magnesium carbonate, dibasic calcium phosphate, tribasic calcium phosphate, magnesium phosphate, silica, zeolite, sodium metaphosphate, aluminum hydroxide, magnesium hydroxide, calcium pyrophosphate, red iron oxide, calcium sulfate, and silicic anhydride.

[0025] Examples of alcohols include ethyl alcohol, lauryl alcohol, and myristyl alcohol. Examples of thickeners include sodium polyacrylate, carrageenan, sodium carboxymethylcellulose, sodium alginate, xanthan gum, hydroxyethyl cellulose, hydroxypropylmethyl cellulose, methyl cellulose, and propylene glycol alginate.

[0026] Examples of medicinal ingredients include fluorides such as sodium monofluorophosphate, sodium fluoride, stannous fluoride, and strontium fluoride, condensed phosphates such as sodium pyrophosphate and sodium polyphosphate, phosphates such as sodium monohydrogen phosphate and trisodium phosphate, vitamins such as ascorbic acid, sodium ascorbate, pyridoxine hydrochloride, and tocopherol acetate, glucanase enzymes such as dextranase and mutanase, proteases, and degradative enzymes such as lysozyme, inorganic salts such as zinc chloride, zinc citrate, strontium chloride, and potassium nitrate, chelating compounds such as chlorophyll and glycerophosphate, fat-dissolving polyethylene glycol, sodium chloride, aluminum lactate, and strontium chloride.

[0027] Examples of colorants include legally designated dyes such as Green No. 1, Blue No. 1, and Yellow No. 4, and titanium oxide. Examples of stabilizers include sodium edetate, sodium thiosulfate, sodium sulfite, calcium lactate, lanolin, triacetin, castor oil, and magnesium sulfate.

[0028] Examples of pH adjusters include citric acid, malic acid, lactic acid, tartaric acid, acetic acid, phosphoric acid, pyrophosphoric acid, glycerophosphoric acid, and various salts thereof such as potassium salts, sodium salts, and ammonium salts, sodium hydroxide, etc. The oral composition is preferably formulated with a pH adjuster to adjust the pH to a range of 4 or more and 9 or less, particularly 5 or more and 7 or less.

[0029] <Action and Effects> The operation of this embodiment will be described. Streptococcus oralis (hereinafter referred to as "S. oralis") is one type of oral flora. It has been found that the protease activity of gingipain produced by gingivalis is inhibited by adding the oral composition of this embodiment to a mixture of S. oralis cells and a culture solution of gingivalis. This is thought to be because the oral composition activates S. oralis, thereby achieving the effect of inhibiting gingipain activity. In other words, it is expected that by holding the oral composition of this embodiment in the mouth, the S. oralis in the oral cavity and specific components contained in the oral composition will interact to inhibit gingipain activity.

[0030] The effects of this embodiment will be described. (1) The oral composition contains yeast extract and a specific citrate or a specific emulsifier, or a specific citrate and a specific emulsifier. The oral composition can inhibit gingipain activity, thereby preventing periodontal disease and suppressing its progression.

[0031] (2) The specific citrate is sodium citrate or ammonium citrate. An oral composition containing sodium citrate or ammonium citrate as the specific citrate can have the effect of inhibiting gingipain activity.

[0032] (3) An oral composition containing a specific citrate and a specific emulsifier has the effect of inhibiting gingipain activity. <Example of change> The above embodiment can be modified as follows: The above embodiment and the following modifications can be combined with each other within the scope of technical compatibility.

[0033] In the above embodiment, it was found that the protease activity of gingipain produced by gingivalis was inhibited by adding the oral composition of the above embodiment to a mixture of S. oralis cells, a normal oral flora, and a culture solution of S. gingivalis. However, the present invention is not limited to this embodiment. Live S. oralis cells may be added to the oral composition. That is, in addition to the S. oralis in the oral cavity interacting with a specific component contained in the oral composition to inhibit gingipain activity, the S. oralis contained in the oral composition may also interact with a specific component contained in the oral composition to inhibit gingipain activity. [Example]

[0034] The oral composition will be described in more detail based on the following examples, but the oral composition is not limited to the configurations described in the examples. Test 1 Gingipain activity was measured by the following method.

[0035] S. gingivalis was cultured in modified GAM medium. The culture solution was diluted with modified GAM medium so that the turbidity at 660 nm was 1.0, and this was used as the S. gingivalis culture solution. S. oralis was cultured in TSB medium. The culture solution was diluted with TSB medium so that the turbidity at 660 nm was 1.0, and this was used as the S. oralis culture solution.

[0036] The S. oralis culture solution was centrifuged and the supernatant was removed to isolate only the S. oralis cells. 100 μL of the sample solution from each of Examples 1 to 4 and Comparative Examples 1 to 7 was added to the S. oralis cells to prepare component-supplemented cells. The types of components contained in each of Examples 1 to 4 and Comparative Examples 1 to 7 are shown in Table 1.

[0037] [Table 1] The concentration of yeast extract in the sample solution containing yeast extract was 0.5% by mass. The concentration of citrate in the sample solution containing citrate was 0.2% by mass. The concentration of anhydrous citric acid in Comparative Example 6 was 0.2% by mass. The concentration of polysorbates in the sample solution containing polysorbates was 0.1% by mass. The remainder of each sample solution was water. The sample solution of Comparative Example 1, i.e., 100 μL of water, was used as the negative control. 100 μL of MRS medium was used as the positive control.

[0038] 100 μL of P. gingivalis culture solution and 100 μL of bacterial cells supplemented with ingredients were added to wells of a 96-well plate and incubated at 37°C under anaerobism for 2 hours. After incubation, 50 μL of the mixture collected from each well was mixed with 50 μL of fluorescent substrate in another well and allowed to react at 37°C for 30 minutes. The fluorescent substrate used was Z-His-Glu-Lys-MCA (Peptide Institute, Inc.). Z-His-Glu-Lys-MCA is benzyloxycarbonyl-L-histidyl-L-glutamyl-L-lysine 4-methylcoumaryl-7-amide (hydrochloride form), a reagent that emits fluorescence upon cleavage by the protease activity of Lys-gingipain (Kgp). The P. gingivalis culture solution contains gingipain produced by P. gingivalis.

[0039] After the reaction, the fluorescence intensity was measured using a fluorescence plate reader (Gemini XPS, manufactured by Molecular Devices) at an excitation wavelength of 380 nm and a fluorescence wavelength of 440 nm. The gingipain activity in the negative control was set to 100%. The gingipain activity in each of Examples 1 to 4 and Comparative Examples 1 to 7 was calculated based on the relative value to the fluorescence intensity of the negative control. The results are shown in Table 1. The gingipain activity of the positive control was 12%.

[0040] Details of the reagents used in this test are as follows: Modified GAM medium: Modified GAM bouillon "Nissui" manufactured by Nissui Pharmaceutical Co., Ltd. TSB medium: Tryptic soy broth manufactured by Becton, Dickinson and Company MRS medium: Lactobacillus MRS broth manufactured by Becton, Dickinson and Company Yeast extract: Becton, Dickinson and Company, BBL Yeast Extract Polysorbate 80: Sigma-Aldrich Sodium citrate: Fujifilm Wako Pure Chemical Industries, Ltd. Ammonium citrate: Fujifilm Wako Pure Chemical Industries, Ltd. <evaluation> It is believed that the smaller the gingipain activity value, the stronger the inhibition of gingipain protease activity. The calculated gingipain activity was evaluated based on the following evaluation criteria.

[0041] Evaluation criteria for gingipain activity ○ (Acceptable): Less than 50%. × (Not acceptable): Greater than 50%.

[0042] As shown in Table 1, the effect of inhibiting gingipain activity was obtained when Example 1, which used a combination of yeast extract and sodium citrate (a citrate salt), was added. The effect of inhibiting gingipain activity was also obtained when Example 2, which used a combination of yeast extract and polysorbate 80 (an emulsifier), was added.

[0043] When Example 3, which uses a combination of yeast extract and ammonium citrate (a citrate salt), was added, the effect of inhibiting gingipain activity was obtained.When Example 4, which uses a combination of yeast extract, ammonium citrate, and polysorbate 80, was added, the effect of inhibiting gingipain activity was also obtained.

[0044] The effect of inhibiting gingipain activity was not obtained when Comparative Examples 2 to 5 were added. That is, when yeast extract, citrate, and polysorbates were used alone, the effect of inhibiting gingipain activity was not obtained.

[0045] Furthermore, when Comparative Example 6, which used a combination of yeast extract and anhydrous citric acid, was added, the effect of inhibiting gingipain activity was not obtained.Also, when Comparative Example 7, which used a combination of ammonium citrate and polysorbate 80, was added, the effect of inhibiting gingipain activity was not obtained.

[0046] The results of Test 1 above show that the effect of inhibiting gingipain activity is an effect that can only be obtained when yeast extract is combined with a specific citrate or a specific emulsifier. It is also shown that the effect of inhibiting gingipain activity can be obtained by further combining a specific emulsifier with the combination of yeast extract and a specific citrate.

[0047] Test 2 In the above-mentioned Test 1, the component-supplemented cells were prepared by adding the sample solution to S. oralis cells. Then, the gingivalis culture solution and the component-supplemented cells were mixed. In Test 2, instead of this step, 100 μL of the gingivalis culture solution and 100 μL of the sample solution were added to the well and incubated at 37°C under anaerobism for 2 hours. In other words, S. oralis cells were not added. The subsequent steps were the same as in Test 1.

[0048] The results of Test 2 are explained below. In the example where a sample solution combining yeast extract and sodium citrate was used, the gingipain activity was 100%. In addition, in the example where a sample solution combining yeast extract and polysorbate 80 was used, the gingipain activity was 106%. In other words, in neither example was the effect of inhibiting gingipain activity obtained. From the results of Tests 1 and 2, it is believed that the effect of inhibiting gingipain activity is obtained by activating S. oralis using a composition combining yeast extract with a specific citrate or a specific emulsifier, or a specific citrate and a specific emulsifier.

Claims

1. An oral composition to be applied in the presence of S. oralis bacteria, An oral composition comprising a yeast extract and at least one selected from citrates and polysorbates, the composition being used to inhibit the protease activity of gingipain.

2. An oral composition comprising a yeast extract and live S. oralis cells, and further comprising at least one selected from citrates and polysorbates, the composition being used to inhibit the protease activity of gingipain.

3. The citrate is sodium citrate or ammonium citrate. The oral composition according to claim 1 or 2.

4. Contains at least one selected from the citrate salts and at least one selected from the polysorbates The oral composition according to any one of claims 1 to 3.

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