Oral components

Abstract

To provide an oral composition having an enhanced effect of improving intraoral flora balance.SOLUTION: An oral composition for improving intraoral flora balance in an oral cavity contains protamine and arginine.SELECTED DRAWING: None

Description

Technical Field

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

Background Art

[0002] Streptococcus oralis (hereinafter also referred to as S. oralis) is known as a kind of non-pathogenic bacteria among oral bacteria. S. oralis belongs to the mitis group of the genus Streptococcus. Other bacterial species belonging to the mitis group are also classified as non-pathogenic bacteria like S. oralis.

[0003] Patent Document 1 discloses an oral composition that selectively inhibits the growth of pathogenic bacteria without affecting the growth of bacterial species belonging to the mitis group of the genus Streptococcus. That is, an oral composition having a selective antibacterial action against pathogenic bacteria in the oral cavity is disclosed. As described in Patent Document 1, it is known that increasing the proportion of non-pathogenic bacteria in the oral flora to improve the oral flora balance is useful for preventing and suppressing the progression of oral diseases.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] By the way, there is a demand for further performance improvement in the effect of improving the flora balance in oral compositions.

Means for Solving the Problems

[0006] The oral composition for solving the above problems contains protamine and arginine, and its purpose is to improve the balance of the oral microbiome. The above oral composition preferably further contains lactose.

[0007] The above oral composition preferably increases the proportion of bacterial species belonging to the mitis group by selectively suppressing the growth of the pathogenic bacterial species among the bacterial species belonging to pathogenic bacteria in the oral cavity and the bacterial species belonging to the mitis group of the genus Streptococcus, which are non-pathogenic bacteria in the oral cavity. [Effects of the Invention]

[0008] The oral composition of the present invention can improve the effect of improving the balance of the bacterial flora in the oral cavity. [Modes for carrying out the invention]

[0009] The following describes one embodiment of the oral composition. The oral composition of this embodiment contains protamine and arginine. The oral composition is used to improve the balance of the bacterial flora in the oral cavity.

[0010] <Protamine> The protamine contained in the oral composition will be described below. Protamine is a basic polypeptide. Protamine is one of the components contained in shirako protein extract. Protamine can be obtained, for example, from the milt of fish. Examples of fish include fish of the Salmonidae family in the Salmoniformes order and fish of the Clupeidae family in the Clupeformes order. Thus, it is preferable that the protamine is of fish origin.

[0011] The oral composition may contain a protamine derived from one type of fish, or it may contain a combination of protamines derived from two or more types of fish. Preferably, the oral composition contains a protamine derived from salmon milt.

[0012] The method for extracting protamine is not particularly limited, and known methods can be used as appropriate. The protamine contained in the oral composition may be purified after extraction. Protamine may be included in the oral composition as a salt. Examples of protamine salts include hydrochloride and sulfate.

[0013] The protamine content in the oral composition is not particularly limited, but is, for example, 0.01% by mass or more and 0.10% by mass or less. Hereinafter, the content of each component in the oral composition will be expressed using "%" instead of "% by mass". The upper limit of the protamine content is preferably 0.075%, and more preferably 0.05%. The lower limit of the protamine content is preferably 0.03%, and more preferably 0.05%.

[0014] By having a protamine content within the above-mentioned range and an arginine content within the range described below, the effect of improving the balance of the bacterial flora in the oral cavity can be further enhanced.

[0015] <Arginine> The oral composition contains arginine, a type of amino acid. Specific examples of arginine are not particularly limited, but include, for example, L-arginine and dihydroxypropylarginine. These arginines may also be in the form of salts, such as hydrochloride salts.

[0016] The oral composition may contain, in addition to arginine, one or more other amino acids in combination. The arginine content in the oral composition is not particularly limited, but is, for example, 0.5% to 1.5%. The upper limit of the above content is preferably 1.25%, more preferably 1.0%. The lower limit of the above content is preferably 0.75%, more preferably 1.0%.

[0017] <sugar> The oral composition preferably contains sugar. The type of sugar is not particularly limited, and examples thereof include monosaccharides, disaccharides, oligosaccharides having three or more saccharide units, and sugar alcohols.

[0018] Examples of the monosaccharide include galactose, mannose, arabinose, and their hydrates. Examples of the disaccharide include maltose, sucrose, lactose, and their hydrates.

[0019] Examples of the oligosaccharide include raffinose, galactooligosaccharide, soy oligosaccharide, lactulose oligosaccharide, xylooligosaccharide, gentiobiose oligosaccharide, maltooligosaccharide, cyclic oligosaccharide, milk oligosaccharide, and their hydrates.

[0020] Examples of the sugar alcohol include those obtained by industrially reducing the above-mentioned monosaccharide, disaccharide, or oligosaccharide, and examples thereof include sorbitol, lactitol, reduced isomaltulose (Palatinit (registered trademark)), trehalose, pentaerythritol, dipentaerythritol, tripentaerythritol, arabitol, ribitol, mannitol, threitol, glycerol, talitol, gaulitol, altitol, dulcitol, iditol, inositol, maltitol, isomaltitol, lactitol, turanitol.

[0021] The oral composition may contain only one of the above sugars alone, or may contain a combination of two or more thereof. In addition, the oral composition particularly preferably contains lactose as the sugar.

[0022] The sugar content in the oral composition is not particularly limited, but for example, it is 1.0% or more and 4.0% or less. The upper limit value of the above content is preferably 3.0%, more preferably 2.0%. The lower limit value of the above content is preferably 1.5%, more preferably 2.0%. By the sugar content being within the above numerical range, the effect of improving the bacterial flora balance in the oral cavity can be further enhanced.

[0023] 〈Application form, use, and dosage form〉 The application form of the oral composition is not particularly limited, and for example, it can be used as a pharmaceutical product, quasi-drug, or cosmetic. As the use of the oral composition, known ones can be appropriately adopted. For example, chewing agents, orally dissolving agents, orally disintegrating agents, tongue care agents, oral cooling agents, dentifrices, mouth rinses, gargles, liquid dentifrices, biofilm dispersants, halitosis preventives, gum massagers, oral wetting agents, tongue coating removers, oral coatings, oral bactericides, throat bactericides, oral and throat agents, periodontal disease therapeutics, denture adhesives, denture coating agents, denture stabilizers, denture preservatives, denture cleaners, implant care agents, etc. can be mentioned as uses.

[0024] 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, pastas, sprays, gels, liquids, suspensions, gums, etc.

[0025] The type of water used as the solvent is not particularly limited, and for example, distilled water, pure water, ultrapure water, purified water, tap water, etc. can be used. The type of alcohol used as the solvent is not particularly limited, and for example, ethanol can be used. Water and alcohol can also be mixed and used.

[0026] When the oral composition is configured in a liquid state, the content of the solvent such as water is not particularly limited, but it 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.

[0027] <Other ingredients> Oral compositions may contain other components besides those mentioned above, depending on the intended use, form, and application. Examples of other components include antibacterial agents, anti-inflammatory agents, fragrances, humectants, abrasives, alcohols, thickeners, sweeteners, medicinal components, colorants, stabilizers, and pH adjusters. Other components that are known to be incorporated into oral compositions may be used. Oral compositions may contain only one of the above-mentioned other components individually, or they may contain two or more in combination.

[0028] Examples of antibacterial agents include cetylpyridinium chloride, parabens, sodium benzoate, triclosan, chlorhexidine hydrochloride, isopropylmethylphenol, benzalkonium chloride, benzethonium chloride, and hinokitiol.

[0029] Examples of anti-inflammatory agents include glycyrrhizinate, tranexamic acid, and ε-aminocaproic acid. Furthermore, it is undesirable for oral compositions to contain components that exhibit a non-selective action, such as killing both pathogenic and non-pathogenic bacteria. Even if an oral composition contains such components, it is preferable that the amount is extremely small, so as to have little effect on the selective antibacterial action of the oral composition.

[0030] Examples of fragrances include anethole, eugenol, carvone, wintergreen, methyl salicylate, thymol, clove oil, sage oil, ocimene oil, and citronellol.

[0031] Examples of abrasives include calcium carbonate, magnesium carbonate, dicalcium phosphate, tricalcium phosphate, magnesium phosphate, silica, zeolite, sodium metaphosphate, aluminum hydroxide, magnesium hydroxide, calcium pyrophosphate, red iron oxide, calcium sulfate, and anhydrous silicic acid.

[0032] Examples of alcohols include ethyl alcohol, lauryl alcohol, and myristyl alcohol. Examples of thickening agents include sodium polyacrylate, carrageenan, sodium carboxymethylcellulose, sodium alginate, xanthan gum, hydroxyethylcellulose, hydroxypropylmethylcellulose, methylcellulose, and propylene glycol alginate.

[0033] 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; degrading enzymes such as proteases and lysozyme; inorganic salts such as zinc chloride, zinc citrate, strontium chloride, and potassium nitrate; chelating compounds such as chlorophyll and glycerophosphate; lipid-dissolving polyethylene glycol, sodium chloride, aluminum lactate, and strontium chloride.

[0034] Examples of colorants include legally approved pigments such as Green No. 1, Blue No. 1, and Yellow No. 4, as well as titanium dioxide. Examples of stabilizers include sodium edetate, sodium thiosulfate, sodium sulfite, calcium lactate, lanolin, triacetin, castor oil, and magnesium sulfate.

[0035] Examples of pH adjusting agents include citric acid, malic acid, lactic acid, tartaric acid, acetic acid, phosphoric acid, pyrophosphate, glycerophosphate, and various salts thereof such as potassium salts, sodium salts, and ammonium salts, as well as sodium hydroxide. It is preferable that the oral composition is adjusted to have a pH of 4 to 9, and particularly 5 to 7, by incorporating a pH adjusting agent.

[0036] <Mechanism of Action and Effects> The operation of this embodiment will now be described. The effect of oral compositions on improving the balance of the oral microbiota is exerted through selective antibacterial action against pathogenic bacteria in the oral cavity. For example, oral compositions selectively suppress the growth of pathogenic bacteria among species belonging to the pathogenic bacteria in the oral cavity and species belonging to the mitis group of the Streptococcus genus, which are non-pathogenic bacteria in the oral cavity. As a result, oral compositions increase the proportion of species belonging to the mitis group.

[0037] Oral compositions can also increase the proportion of bacterial species belonging to the mitis group by selectively promoting the growth of S. oralis. In other words, oral compositions can improve the balance of the oral microbiota by selectively suppressing the growth of pathogenic bacteria while promoting the growth of S. oralis.

[0038] Oral pathogenic bacteria include caries pathogens and periodontal pathogens. An example of a caries pathogen is Streptococcus mutans, which belongs to the mutans group of the genus Streptococcus. Examples of periodontal pathogens include Porphyromonas gingivalis and Fusobacterium nucleatum. Hereafter, Streptococcus mutans will be abbreviated as S.mutans, Porphyromonas gingivalis as P.gingivalis, and Fusobacterium nucleatum as F.nucleatum.

[0039] The mitis group of the Streptococcus genus, which are non-pathogenic bacteria found in the oral cavity, includes S. oralis, Streptococcus sanguinis, Streptococcus gordonii, and Streptococcus mitis. Hereafter, Streptococcus sanguinis will be abbreviated as S. sanguinis, Streptococcus gordonii as S. gordonii, and Streptococcus mitis as S. mitis.

[0040] The oral composition can, for example, inhibit the growth of S. mutans. The oral composition can also inhibit the growth of P. gingivalis. The oral composition can also inhibit the growth of F. nucleatum.

[0041] Oral compositions can, for example, increase the proportion of S. oralis. Oral compositions can also increase the proportion of S. sanguinis. Oral compositions can also increase the proportion of S. mitis.

[0042] The effects of this embodiment will now be explained. (1) The oral composition contains protamine and arginine. This gives the oral composition a selective antibacterial effect against pathogenic bacteria in the oral cavity. The oral composition can suitably increase the proportion of bacterial species belonging to the mitis group in the oral microbiota. By suitably increasing the proportion of bacterial species belonging to the mitis group, the oral composition can suitably improve the balance of the oral microbiota. In other words, the oral composition can enhance the effect of improving the balance of the oral microbiota.

[0043] (2) Oral compositions further containing lactose can selectively promote the growth of S. oralis. That is, oral compositions can increase the proportion of bacterial species belonging to the mitis group by selectively suppressing the growth of pathogenic bacteria while promoting the growth of S. oralis. Therefore, the effect of improving the balance of the bacterial flora in the oral cavity can be further enhanced.

[0044] (3) Oral compositions that improve the oral microbiota by adjusting the balance of the oral microbiota can be used to prevent oral diseases and inhibit the progression of oral diseases. Specifically, they can be used to prevent dental caries, inhibit the progression of dental caries, prevent periodontal disease, and inhibit the progression of periodontal disease. [Examples]

[0045] The oral compositions will be described in more detail based on the following examples. However, the oral compositions are not limited to those described in the Examples section. <Culture solution> Culture solutions were prepared by culturing each bacterial species in a suitable medium. The bacterial species used were as follows: Non-pathogenic bacteria included S. oralis, S. sanguinis, and S. mitis. Pathogenic bacteria included S. mutans, P. gingivalis, and F. nucleatum. BHI medium was used for Streptococcus species, namely S. oralis, S. sanguinis, S. mitis, and S. mutans. Modified GAM medium was used for P. gingivalis. GAM medium was used for F. nucleatum.

[0046] (Test 1) Tables 1 and 2 will be used to explain the antibacterial efficacy test.

[0047] [Table 1] Sample solutions for Comparative Examples 1-3 and Examples 1 and 2, as shown in Table 1, were prepared. In Table 1, "○" indicates the presence of the component. "-" indicates the absence of the component. The content of each component is as follows: Protamine content: 0.05%. Arginine content: 1%. Lactose content: 2%. The remainder of each sample solution is distilled water.

[0048] <Preparation of bacterial suspension> The bacterial suspensions to be used in Experiment 1 were prepared. First, a 2x concentration medium was prepared. Next, the turbidity of the culture medium at a wavelength of 660 nm was measured. The culture medium was diluted with the 2x concentration medium until the turbidity was 0.1 to prepare the bacterial suspension. Bacterial suspensions for each bacterial species were prepared using the culture media of S. oralis, S. mutans, P. gingivalis, and F. nucleatum.

[0049] <Test method for antibacterial efficacy testing> 100 μl of bacterial suspension and 100 μl of sample solution were added to the wells of a 96-well plate and incubated at 37°C under anaerobic conditions for 24 hours. As a control, 100 μl of water was added instead of the sample solution and incubated in the same manner. After incubation, turbidity at a wavelength of 660 nm was measured.

[0050] Based on the measured turbidity, the relative activity of each bacterial species was calculated by setting the control turbidity to 1.00. In other words, a relative activity greater than 1.00 indicates that the growth of the bacterial species is promoted. Conversely, a relative activity less than 1.00 indicates that the growth of the bacterial species is suppressed.

[0051] <Results of antibacterial efficacy test> The calculated relative activity was evaluated according to the following criteria. The results are shown in Table 2. • Criteria for evaluating relative activity in antimicrobial efficacy tests ◎: Relative activity is 1.50 or higher. The growth of the bacterial species is promoted. ○: Relative activity is between 0.90 and 1.50. The bacterial species is growing. △: Relative activity is between 0.50 and 0.90. The growth of the bacterial species is suppressed to some extent. ×: Relative activity is less than 0.50. The growth of the bacterial species is suppressed.

[0052] [Table 2] In Table 2, So, Sm, Pg, and Fn refer to S. oralis, S. mutans, P. gingivalis, and F. nucleatum, respectively. Hereafter, each fungal species may be abbreviated similarly.

[0053] In Example 1, the growth of Sm, Pg, and Fn was suppressed, while the growth of So was not suppressed and grew to some extent. This selective antibacterial effect can be seen from a comparison with the results of Comparative Examples 1 and 2, and is the effect obtained by Example 1, which contains protamine and arginine.

[0054] In Example 2, the growth of Sm, Pg, and Fn was suppressed. Furthermore, the growth of So was promoted. This selective antibacterial and growth-promoting effect is evident from a comparison with the results of Comparative Examples 1-3, and is the effect obtained by Example 2, which further contains lactose in addition to protamine and arginine.

[0055] Examples 1 and 2 demonstrate that the growth of pathogenic bacteria is selectively suppressed among pathogenic bacteria in the oral cavity and non-pathogenic bacteria belonging to the mitis group of the genus Streptococcus. Examples 1 and 2 suggest that the proportion of mitis group species in the oral microbiota can be increased, thereby exhibiting an improvement effect on the oral microbiota.

[0056] (Exam 2) Tables 1 and 3 will be used to explain the sterilization efficacy test. The sample solutions used were those from Examples 1 and 2, and Comparative Examples 1 to 3, as shown in Table 1.

[0057] <Preparation of bacterial suspension> The bacterial suspensions to be used in Experiment 2 were prepared. The turbidity of the culture medium at a wavelength of 660 nm was measured. The culture medium was diluted with phosphate-buffered saline to obtain a bacterial suspension with a turbidity of 1.0. Bacterial suspensions were prepared for each bacterial species using the culture media of S. oralis, S. mutans, P. gingivalis, and F. nucleatum.

[0058] <Test method for sterilization effect test> 20 μl of bacterial suspension and 180 μl of sample solution were added to the wells of a 96-well plate. As a control, 180 μl of water was added instead of the sample solution. After a predetermined contact time, 20 μl of the solution from each well was added to 180 μl of inactivation medium. The contact times were 30 seconds and 3 minutes. Subsequently, the cells were incubated at 37°C under anaerobic conditions for 24 hours. Turbidity was measured, and relative activity was calculated in the same manner as in Test 1 above.

[0059] The inactivated culture medium was prepared by adding Tween® 80 and soy lecithin, both known as inactivators of fungicides, to each culture medium. When a fungicide is added to the inactivated culture medium, the fungicide is inactivated.

[0060] <Results of the sterilization effect test> The calculated relative activity was evaluated according to the following criteria. The results are shown in Table 3. • Criteria for evaluating relative activity in sterilization efficacy tests ◎: Relative activity is 1.50 or higher. The growth of the bacterial species is promoted. ○: Relative activity is between 0.90 and 1.50. The bacterial species is growing. △: Relative activity is between 0.50 and 0.90. The growth of the bacterial species is suppressed to some extent. ×: Relative activity is less than 0.50. The growth of the bacterial species is suppressed.

[0061] [Table 3] In Example 1, the relative activity of Sm and Pg decreased when the contact time was 30 seconds and when the contact time was 3 minutes. From these results, it can be seen that the effect of suppressing the growth of Sm and Pg in Example 1 is obtained when the contact time is 30 seconds or longer.

[0062] In Example 2, the relative activity of Sm decreased when the contact time was 30 seconds and when the contact time was 3 minutes. From these results, it can be seen that the effect of suppressing Sm growth in Example 2 is obtained when the contact time is 30 seconds or longer.

[0063] In Example 2, the relative activity of Pg decreased when the contact time was 3 minutes. From this result, it can be seen that in Example 2, the effect of suppressing Pg growth was not obtained when the contact time was 30 seconds, but was obtained when the contact time was 3 minutes or longer.

[0064] In Examples 1 and 2, the relative activity of Fn decreased when the contact time was 3 minutes. From these results, it can be seen that the effect of suppressing Fn growth is not obtained when the contact time is 30 seconds, but is obtained when the contact time is 3 minutes or longer.

[0065] (Exam 3) Table 4 describes the tests that verified the selectivity of the selective antibacterial activity exhibited by protamine. Specifically, in Test 3, the effect of protamine on bacterial species of the Streptococcus genus, mitis group, was verified.

[0066] <Preparation of bacterial suspension> The bacterial suspension to be used in Experiment 3 was prepared. First, a 2x concentration medium was prepared. Next, the turbidity of the culture medium at a wavelength of 660 nm was measured. The culture medium was diluted with the 2x concentration medium until the turbidity was 0.1 to obtain the bacterial suspension.

[0067] Bacterial suspensions of each species were prepared using culture media of the mitis group (Streptococcus genus) including S. oralis, S. sanguinis, and S. mitis, and the mutans group (Streptococcus genus) including S. mutans. <Testing Method> Protamine was dissolved in distilled water to prepare a dilution series. These dilution series were used as sample solutions for the antibacterial efficacy test, as in Test 1. The bacterial suspension used was the one prepared as described above. The concentration at which bacterial growth was not observed was defined as the minimum inhibitory concentration (MIC). The results are shown in Table 4.

[0068] [Table 4] <Test Results> Regarding protamine, the MIC value for S. oralis was higher than that for S. mutans. This confirms that it selectively inhibits the growth of S. mutans while being less effective at inhibiting the growth of S. oralis. Furthermore, the MIC values ​​for S. sanguinis and S. mitis, not just S. oralis, were higher than those for S. mutans. This confirms that the growth of S. sanguinis and S. mitis, not just S. oralis, is less effectively inhibited. From this, it can be seen that it exhibits selectivity in inhibiting the growth of species in the mitis group.

[0069] An oral composition containing protamine and arginine can be expected to increase the proportion of mitis group cells present. (Reagent details) The details of the reagents used in each test described in the Examples section are as follows:

[0070] BHI medium: Becton, Dickinson and Company, Brain Heart Infusion Broth GAM culture medium: Manufactured by Nissui Pharmaceutical Co., Ltd., GAM broth Modified GAM medium: Manufactured by Nissui Pharmaceutical Co., Ltd., Modified GAM broth Protamine: Manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., protamine sulfate Arginine: L-arginine, manufactured by Sigma-aldrich. Lactose: Manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., lactose monohydrate

Claims

1. It contains protamine and arginine. The protamine content is 0.01% by mass or more and 0.1% by mass or less. An oral composition for improving the balance of the bacterial flora in the oral cavity.

2. The oral composition according to claim 1, further containing lactose.

3. An oral composition according to claim 1 or 2, wherein the composition selectively inhibits the growth of the pathogenic bacteria species, thereby increasing the proportion of the mitis group species, among the pathogenic bacteria species in the oral cavity and the mitis group species of the Streptococcus genus which are non-pathogenic bacteria in the oral cavity, and the pathogenic bacteria species whose growth is selectively inhibited include at least one of Porphyromonas gingivalis and Fusobacterium nucleatum.

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