JP7330692B2 - oral composition - Google Patents

Description

TECHNICAL FIELD The present invention relates to oral compositions and the like, and more particularly to oral compositions and the like containing retinol palmitate.

Periodontal disease progresses due to interactions between bacteria in plaque and responses from the body. Therefore, in order to prevent periodontal disease, it is considered effective to control periodontopathogenic bacteria by physical and chemical removal and to control the host's defense function.

In particular, the periodontal bacterium, Porphyromonas gingivalis (Pg fungus), disrupts intercellular adhesion of the gingival epithelium, allowing virulence factors to enter the tissue. This leads to the development and/or progression of periodontal disease.

Japanese Patent Application Publication No. 2018-090585

A main object of the present invention is to provide an oral composition capable of preventing and/or suppressing periodontal disease.

Conventionally, oral care products focused on sterilization and anti-inflammatory properties, and many of these products retroactively had these effects. On the other hand, gingival epithelial cells function as a physical barrier at the forefront against invasion of periodontal disease bacteria, and can play an important role for prevention and/or suppression of periodontal disease. As mentioned above, P.I. g. Strengthening the intercellular adhesion of gingival epithelial cells because bacteria destroy the intercellular adhesion of gingival epithelium and invasion of pathogenic factors into tissues can contribute to the onset and/or progression of periodontal disease. Therefore, it is expected that periodontal disease can be prevented and/or suppressed.

Accordingly, the present inventors searched for a component that exerts the effect of strengthening the intercellular adhesion of gingival epithelial cells, and found that retinol palmitate exerts the effect, and made further improvements.

The invention includes, for example, the subject matter described in the following sections.
Section 1.
An anti-periodontal oral composition containing retinol palmitate.
Section 2.
Item 1. The oral cavity composition according to Item 1, containing 0.0001 to 1% by mass of retinol palmitate.
Item 3.
An oral composition containing 0.0001 to 1% by mass of retinol palmitate.
Section 4.
Item 1. The oral cavity composition according to Item 1, containing 1 to 10,000 IU of retinol palmitate.
Item 5.
An oral composition containing 1 to 10,000 IU of retinol palmitate.
Item 6.
Item 6. The composition for oral cavity according to any one of Items 1 to 5, which is for enhancing adhesion between gingival epithelial cells.
Item 7.
Item 7. The oral composition according to any one of Items 1 to 6, further comprising cetylpyridinium chloride.
Item 8.
Item 8. The oral composition according to Item 7, containing 0.01 to 0.5% by mass of cetylpyridinium chloride.
Item 9.
A gingival epithelial intercellular adhesion enhancer containing retinol palmitate.

Periodontal disease can be prevented and/or suppressed by enhancing intercellular adhesion of gingival epithelial cells.

The outline of the case which examined whether the intercellular adhesion destruction inhibitory effect is acquired by the retinol palmitate of each density|concentration by the amount of FITC permeate|transmitting between Epi4 cells is shown. Fig. 2 shows the results of examining whether or not each concentration of retinol palmitate has an inhibitory effect on intercellular adhesion disruption based on the amount of FITC permeating between Epi4 cells. The result of examining whether various vitamin compounds have the effect of inhibiting the destruction of intercellular adhesion is shown by the amount of FITC permeating between Epi4 cells. The results of an examination using WST-1 to determine whether each concentration of retinol palmitate causes cytotoxicity are shown.

Each embodiment included in the present invention will be described in further detail below. The present invention preferably includes, but is not limited to, oral compositions, uses thereof, and the like, and includes everything disclosed herein and recognized by those skilled in the art.

The oral compositions encompassed by the present invention contain retinol palmitate. The oral composition included in the present invention is sometimes referred to as "the oral composition of the present invention".

Retinol palmitate is an ester of palmitic acid and retinol, and is a kind of vitamin A. The structural formula of retinol palmitate is shown below.

In the oral cavity composition of the present invention, retinol palmitate is preferably contained in an amount of 0.0001% by mass (w/w) or more, and the upper limit is not particularly limited, but for example, it is contained in an amount of about 0.0001 to 1% by mass. is more preferable. The lower limit of the range is, for example, about 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, or 0.001% by mass. good. The upper limit of the range is, for example, 0.5, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, or 0 It may be about 0.01% by mass. In terms of international units (IU), the content is preferably 1 IU or more, and although the upper limit is not particularly limited, it is more preferably about 1 to 10000 IU. The lower limit of the range may be, for example, about 2, 3, 4, 5, 6, 7, 8, 9, or 10 IU. The upper limit of the range may be, for example, about 5000, 1000, 900, 800, 700, 600, 500, 400, 300, 200, or 100 IU.

In addition, since the composition for oral cavity of the present invention contains retinol palmitate, it has the effect of enhancing intercellular adhesion of gingival epithelial cells when applied in the oral cavity. Therefore, the composition for oral cavity of the present invention can be preferably used for enhancing adhesion between gingival epithelial cells. It can also be preferably used for suppressing the disruption of intercellular adhesion by Porphyromonas gingivalis (P.g. fungus). Furthermore, since such effects are exhibited, the composition for oral cavity of the present invention can be preferably used as an anti-periodontal composition for oral cavity.

In addition, since the oral composition of the present invention has the effect of enhancing intercellular adhesion of gingival epithelial cells, P. g. In the oral cavity infected with bacteria, not only the effect of suppressing the progression of periodontal disease, but also P. g. It also has the effect of preventing periodontal disease in the oral cavity that is not infected with bacteria. For this reason, P.I. g. To suppress the progression of periodontal disease in people infected with bacteria in the oral cavity, and/or P. g. It can be preferably used for the prevention of periodontal disease in non-infected oral cavity bacteria.

Moreover, the oral composition of the present invention preferably further contains cetylpyridinium chloride. When cetylpyridinium chloride is contained, it is preferably contained in an amount of 0.01 to 0.5% by mass. The lower limit may be 0.02, 0.03, 0.04, or 0.05 wt%, and the upper limit may be 0.4, 0.3, 0.2, or 0.1 wt%. There may be. For example, it is more preferable to contain about 0.02 to 0.3% by mass.

Cetylpyridinium chloride is a bactericidal agent that is widely used in the field of oral compositions, and by containing it, the oral composition of the present invention also exhibits a bactericidal effect (especially a Pg fungicidal effect). Therefore, it is preferable.

The oral composition of the present invention can be a solid composition or a liquid composition. The oral cavity composition can be used, for example, as a drug or quasi-drug. The form of the oral composition of the present invention is not particularly limited. It can be in the form (dosage form) of toothpaste, gum, and the like. Among them, mouthwashes, liquid dentifrices, toothpastes, ointments, pastes, liquids and gels are preferred.

The composition for oral cavity of the present invention may further contain one or more optional ingredients that can be blended in the composition for oral cavity within a range that does not impair the effects of the present invention.

For example, nonionic surfactants, anionic surfactants or amphoteric surfactants can be blended as surfactants. Specifically, for example, nonionic surfactants include sugar fatty acid esters such as sucrose fatty acid esters, maltose fatty acid esters, and lactose fatty acid esters; fatty acid alkanolamides; sorbitan fatty acid esters; fatty acid monoglycerides; ~10, polyoxyethylene alkyl ethers in which the number of carbon atoms in the alkyl group is 13 to 15; polyoxyethylene alkylphenyl ethers in which the polyoxyethylene addition coefficient is 10 to 18, and the number of carbon atoms in the alkyl group is 9; diethyl sebacate; polyoxyethylene hydrogenated castor oil; fatty acid polyoxyethylene sorbitan; and the like. Examples of anionic surfactants include sulfuric acid ester salts such as sodium lauryl sulfate and sodium polyoxyethylene lauryl ether sulfate; sulfosuccinates such as sodium lauryl sulfosuccinate and sodium polyoxyethylene lauryl ether sulfosuccinate; sodium cocoyl sarcosinate and lauroyl methylalanine. acyl amino acid salts such as sodium; cocoyl methyl taurate sodium; Examples of amphoteric surfactants include betaine acetate surfactants such as betaine lauryldimethylaminoacetate and betaine coconut oil fatty acid amidopropyldimethylaminoacetate; imidazoline surfactants such as sodium N-cocoyl-N-carboxymethyl-N-hydroxyethylethylenediamine. Active agent: amino acid-type active agent such as N-lauryldiaminoethylglycine, and the like. These surfactants can be blended singly or in combination of two or more. The blending amount thereof is usually 0.1 to 5% by mass based on the total amount of the composition.

Flavoring agents such as menthol, carboxylic acid, anethole, eugenol, methyl salicylate, limonene, ocimene, n-decyl alcohol, citronol, α-terpineol, methyl acetate, citronenyl acetate, methyl eugenol, cineol, Linalool, ethyl linalool, thymol, spearmint oil, peppermint oil, lemon oil, orange oil, sage oil, rosemary oil, cinnamon oil, perilla oil, wintergreen oil, clove oil, eucalyptus oil, pimento oil, d-camphor, Perfumes such as d-borneol, fennel oil, cinnamon oil, cinnamaldehyde, peppermint oil and vanillin can be used. These may be blended singly or in combination of two or more in an amount of, for example, 0.001 to 1.5% by mass based on the total amount of the composition.

Sweeteners that can be used include saccharin sodium, acesulfame potassium, stevioside, neohesperidyl dihydrochalcone, perillartine, thaumatin, asparatylphenylalanylmethyl ester, p-methoxycinnamic aldehyde, and the like. These can be blended, for example, in an amount of 0.01 to 1% by mass relative to the total amount of the composition.

Furthermore, as wetting agents, sorbit, ethylene glycol, propylene glycol, glycerin, 1,3-butylene glycol, polypropylene glycol, xylit, maltit, lactit, polyoxyethylene glycol, etc. may be blended alone or in combination of two or more. can.

As preservatives, parabens such as methylparaben, ethylparaben, propylparaben and butylparaben, sodium benzoate, phenoxyethanol, alkyldiaminoethylglycine hydrochloride, and the like can be added.

As a coloring agent, legal dyes such as Blue No. 1, Yellow No. 4, Red No. 202 and Green No. 3, mineral dyes such as ultramarine blue, enhanced ultramarine blue and Prussian blue, titanium oxide, and the like may be blended.

As a pH adjuster, citric acid, phosphoric acid, malic acid, pyrophosphoric acid, lactic acid, tartaric acid, glycerophosphoric acid, acetic acid, nitric acid, or chemically possible salts thereof, sodium hydroxide, or the like may be added. These may be blended singly or in combination of two or more so that the pH of the composition is in the range of 4-8, preferably 5-7. The blending amount of the pH adjuster may be, for example, 0.01 to 2% by weight.

The oral composition of the present invention contains not only cetylpyridinium chloride, but also vitamin E compounds such as dl-α-tocopherol acetate, tocopherol succinate, or tocopherol nicotinate and amphoteric compounds such as dodecyldiaminoethylglycine as active ingredients. Fungicides, nonionic fungicides such as triclosan, isopropylmethylphenol, hinokitiol, anionic fungicides such as sodium lauroyl sarcosinate, cationic fungicides such as chlorhexidine hydrochloride, benzalkonium chloride, benzethonium chloride, dextranase, amylase , enzymes such as protease, mutanase, lysozyme, lytic enzyme (retech enzyme), alkali metal monofluorophosphates such as sodium monofluorophosphate and potassium monofluorophosphate, fluorides such as sodium fluoride and stannous fluoride , tranexamic acid, epsilon aminocaproic acid, aluminum chlorohydroxyallantoin, dihydrocholesterol, glycyrrhetinic acid, glycyrrhizic acid, sodium copper chlorophyllin, glycerophosphate, chlorophyll, sodium chloride, calopeptide, allantoin, carbazochrome, hinokitiol, potassium nitrate, palatinit, etc. Or it can mix|blend in combination of 2 or more types. These medicinal ingredients may be used in combination with cetylpyridinium chloride, or may be added to the oral composition of the present invention that does not contain cetylpyridinium chloride.

As a base, alcohols, silicon, apatite, white petrolatum, paraffin, liquid paraffin, microcrystalline wax, squalane, plastibase, etc. can be added.

In addition, the oral composition of the present invention can be prepared by a known method or a method easily conceived from known methods. For example, it can be prepared by appropriately mixing retinol palmitate and, if necessary, other components (especially cetylpyridinium chloride).

In this specification, the term "comprising" includes "consisting essentially of" and "consisting of." In addition, the present invention encompasses all arbitrary combinations of the constituent elements described herein.

Also, the various characteristics (property, structure, function, etc.) described for each of the embodiments of the invention described above may be combined in any way to identify subject matter encompassed by the invention. That is, the invention encompasses all subject matter consisting of any and all possible combinations of the features described herein.

The present invention will be described in more detail below, but the present invention is not limited to the following examples.

Investigation of inhibitory effect of retinol palmitate on disruption of intercellular adhesion Gingival epithelial cell line Epi4 cells were cultured on a transwell until confluent. The transwell is designed to be suspended with the insert in the middle of the well and is a tool that allows cell permeability assessment.

A cell medium containing 2.5% DMSO containing retinol palmitate was also prepared. For the preparation, a commercially available reagent, retinol palmitate, is used, and the reagent is 0.1% by mass, 0.05% by mass, 0.01% by mass, 0.005% by mass, or 0.001% by mass in the medium. included.

300 μl of the prepared medium containing retinol palmitate at each concentration was added to Epi4 cells cultured on a transwell (culture medium removed) from the upper surface and incubated for 1 hour at 37° C. under 5% CO ₂ concentration.

In addition, pre-cultured P. P. gingivalis W83 was collected by centrifugation at 10000 rpm and the concentration was adjusted to OD600=0.5. After the above incubation, the Epi4 cells on the transwell were coated with each adjusted concentration of P.I. g. 100 μl of the bacterial solution was added and incubated for 2 hours at 37° C. under a CO ₂ concentration of around 5%.

The retinol palmitate reagent used in the study was 215000 IU/g. Therefore, the above 0.1% by mass, 0.05% by mass, 0.01% by mass, 0.005% by mass, or 0.001% by mass is 215 IU and 107.5 IU, respectively, in international units (IU) , 21.5 IU, 10.75 IU, and 2.15 IU. The formula for converting the international unit (IU) of retinol palmitate into mass is 1 IU=0.55 μg. Therefore, the retinol palmitate reagent contains retinol palmitate at a concentration of 215000 IU/g×0.55 μg=118.25 mg/g. Furthermore, since the density of commercially available retinol palmitate reagents is usually 0.9-0.95 mg/ml, 118.25 mg/g×1/0.9-0.95=124-130 mg/ml, the palmitin The acid retinol reagent will contain retinol palmitate. Further, for example, in the above-described study in which a medium containing 0.1% by mass of retinol palmitate reagent was prepared, the initial concentration was 900 to 950×118.25=106.4 to 112.3 μg/ml. about 0.0106 to 0.0112% by mass (w/w)% concentration. Similarly, in studies conducted by preparing media containing 0.05% by mass, 0.01% by mass, 0.005% by mass, or 0.001% by mass of the retinol palmitate reagent, the initial concentration was 0 .1/2, 1/10, 1/20, or 1/100 of 1% by mass. Further, the final concentration (concentration after addition of P.g bacteria) is 3/4 of the concentration value.

After incubation, the medium was removed, and after washing with PBS, FITC-dextran (4 kDa) adjusted to 1 mg/ml was added from the top of the transwell and incubated at 37° C. for 1 hour. FITC-dextran that passed through the transwell was recovered, and fluorescence intensity (excitation light: 490 nm, emission light: 520 nm) was measured using a fluorescence plate reader (Gemini XPS). The permeation rate of FITC at a retinol palmitate concentration of 0 IU was converted to 100%, and the transmittance when treated with retinol palmitate of each concentration was calculated. It can be said that the lower the permeation rate, the higher the intercellular adhesion destruction inhibitory effect. FITC is an abbreviation for Fluoresceinisothiocyanate isomer-I.

The outline of the above experiment is shown in FIG. FIG. 2 shows the study results (FITC transmittance calculated from the FITC transmittance amount). In FIG. 2, the concentration of retinol palmitate is represented by the final concentration (IU).

Investigation of the inhibitory effect of various vitamins on the destruction of intercellular adhesion Various vitamin compounds (specific compound names are shown in FIG. 3) reagents (final concentration 0.0075%) were used, and the concentration was such that OD600 = 0.1. adjusted P. In the same manner as described above, except that the culture solution of the g bacteria was used, it was examined whether various vitamins have the effect of inhibiting the destruction of intercellular adhesion. The results are shown in FIG. Among various vitamin compounds, it was confirmed that retinol palmitate exhibits a particularly excellent effect of inhibiting the destruction of intercellular adhesion.

Examination of cytotoxicity of retinol palmitate Gingival epithelial cell line Epi4 cells were cultured in 96 wells until confluent.
Cell culture medium containing 2.5% DMSO containing each concentration of retinol palmitate was added and incubated at 37° C. for 3 hours.

WST-1, which is a reagent for cell proliferation activity, was added and incubated at 37°C for 40 minutes. WST-1 is a tetrazolium salt compound represented by the following structural formula, and is used for proliferative capacity measurement in which the number of viable cells is analyzed by decomposition of the salt.

After incubation, the absorbance (OD (450 nm)) was measured with an absorbance plate reader (xMark microplate reader). The absorbance at a retinol palmitate concentration of 0 IU was converted to 100%, and the cell survival rate when treated with each concentration of retinol palmitate was calculated. The results are shown in FIG. No cytotoxicity was observed with retinol palmitate.

Investigation of bactericidal activity of retinol palmitateP . G bacteria (P. gingivalis W83) were cultured in GAM medium, and O. g bacteria were cultured in GAM medium. D(600) was adjusted to 1.0. A drug (retinol palmitate, cetylpyridinium chloride (CPC), or a combination thereof) adjusted to each concentration was mixed with the adjusted bacterial solution and allowed to stand for 3 minutes. After 3 minutes, 10 times this amount of fungicide inactivation medium (0.07% lecithin + 0.5% Tween 80) was added to stop the bactericidal effect, and cultured under anaerobic conditions at 37°C for 24 hours. After culturing, the absorbance (OD (600)) of the medium was measured with an absorption plate reader (xMark microplate reader), and the bactericidal effect was determined from the presence or absence of turbidity. It can be said that the lower the absorbance, the higher the bactericidal effect. The results are shown in Table 1. No bactericidal activity was observed in retinol palmitate, and no effect of inhibiting the bactericidal effect of CPC was observed.

Retinol palmitate does not exhibit cytotoxicity and bactericidal activity. It was found that this was not because the disruption of intercellular adhesion was suppressed due to damage to g bacteria, but because the disruption of intercellular adhesion was suppressed due to enhanced intercellular adhesion of Epi4 cells.

Claims (5)

An anti-periodontal oral composition containing retinol palmitate and cetylpyridinium chloride, except oral compositions containing panthenol or pantothenate and oral compositions containing beta-carotene . The oral composition according to claim 1, containing 0.0001 to 1% by mass of retinol palmitate. The oral composition according to claim 1, containing 1 to 10,000 IU of retinol palmitate. The composition for oral cavity according to any one of claims 1 to 3, which is for enhancing adhesion between gingival epithelial cells. The oral composition according to any one of claims 1 to 4, containing 0.01 to 0.5% by mass of cetylpyridinium chloride.