JP2020097534A - Oral composition - Google Patents

Abstract

To provide an oral composition that can prevent and/or inhibit periodontitis.SOLUTION: An anti-periodontitis oral composition contains retinol palmitate.SELECTED DRAWING: None

Description

The present invention relates to an oral composition and the like, and more particularly to an oral composition and the like containing retinol palmitate.

Periodontal disease progresses due to the interaction between bacteria in the plaque and the response on the living body side. Therefore, in order to prevent periodontal disease, it is considered effective to control periodontopathic bacteria by physical and chemical removal and to control host side defense function.

In particular, the periodontal disease bacterium, Porphyromonas gingivalis (P. g. bacterium), destroys the intercellular adhesion of the gingival epithelium, and the pathogenic factor penetrates into the tissue. This leads to the onset and/or progression of periodontal disease.

JP-A-2018-090585

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

Heretofore, many oral care products have focused on sterilization and anti-inflammatory and have retrospective effects. On the other hand, the gingival epithelial cells function as a physical barrier at the forefront against the invasion of periodontal disease bacteria, and can play an important role for preventing and/or suppressing periodontal disease. As described above, P. g. Bacteria disrupt intercellular adhesion of gingival epithelium, and invasion of pathogenic factors into tissues may contribute to the onset and/or progression of periodontal disease, thus enhancing intercellular adhesion of gingival epithelial cells Therefore, it is expected that the periodontal disease can be prevented and/or suppressed.

Then, as a result of searching for a component having an effect of enhancing the intercellular adhesion of gingival epithelial cells, the present inventors have found that retinol palmitate has the effect, and further improved it.

The present invention includes the subject matter described in the following sections, for example.
Item 1.
An anti-periodontal disease oral composition containing retinol palmitate.
Item 2.
Item 2. The oral composition according to Item 1, containing 0.0001 to 1 mass% of retinol palmitate.
Item 3.
A composition for oral cavity containing 0.0001 to 1% by mass of retinol palmitate.
Item 4.
Item 2. The oral 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 7. The oral composition 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 containing cetylpyridinium chloride.
Item 8.
Item 8. The oral composition according to Item 7, which contains 0.01 to 0.5% by mass of cetylpyridinium chloride.
Item 9.
A gingival epithelial cell-cell adhesion enhancer containing retinol palmitate.

By enhancing intercellular adhesion of gingival epithelial cells, periodontal disease can be prevented and/or suppressed.

The outline of the case in which whether each concentration of retinol palmitate has an inhibitory effect on cell-cell adhesion disruption is examined by the amount of FITC penetrating between Epi4 cells is shown. The results of examining whether or not the retinol palmitate at each concentration has an effect of suppressing intercellular adhesion disruption by the amount of FITC penetrating between Epi4 cells are shown. The results of examining whether or not the intercellular adhesion disruption inhibitory effect is obtained by various vitamin compounds by the amount of FITC that penetrates between Epi4 cells are shown. The result of having investigated using WST-1 whether cytotoxicity is caused by each concentration of retinol palmitate is shown.

Hereinafter, each embodiment included in the present invention will be described in more detail. The present invention preferably includes oral compositions, uses thereof, and the like, but is not limited thereto, and the present invention includes all disclosed in the present specification and recognizable by those skilled in the art.

The oral composition included in the present invention contains retinol palmitate. The oral composition included in the present invention may be referred to as “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 composition for oral cavity of the present invention, the retinol palmitate is preferably contained in an amount of 0.0001 mass (w/w)% or more, and the upper limit is not particularly limited, but is, for example, about 0.0001 to 1 mass%. Is more preferable. The lower limit of the range may be, 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. If it is described in international units (IU), it is preferably contained at 1 IU or more, and the upper limit is not particularly limited, but it is more preferably contained at 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, the oral composition of the present invention, by containing retinol palmitate, has the effect of enhancing the intercellular adhesion of gingival epithelial cells when applied in the oral cavity. Therefore, the oral composition of the present invention can be preferably used for enhancing adhesion between gingival epithelial cells. Further, it can also be preferably used for suppressing the destruction of cell-cell adhesion by Porphyromonas gingivalis (P. g. bacterium). Furthermore, since it exhibits such effects, the oral cavity composition of the present invention can be preferably used as an anti-periodontal disease oral cavity composition.

The oral composition of the present invention has the effect of strengthening the intercellular adhesion of gingival epithelial cells. 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 not infected with bacteria. Therefore, P. g. Bacterial infection for suppressing the periodontal disease progression of the infected person, and/or P. g. It can be preferably used for the prevention of periodontal disease in non-infected persons with oral cavity.

The oral composition of the present invention preferably further contains cetylpyridinium chloride. When containing cetylpyridinium chloride, 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% by weight, and the upper limit is 0.4, 0.3, 0.2, or 0.1% by weight. It may be. For example, it is more preferable to contain 0.02 to 0.3% by mass.

In addition, cetylpyridinium chloride is a bactericide widely used in the field of oral composition, and the oral composition of the present invention also has a bactericidal effect (particularly P. g. bactericidal effect) by containing it. This is preferable.

The oral composition of the present invention may be a solid composition or a liquid composition. The composition for oral cavity can be used, for example, as a drug or a quasi drug. Further, the form of the composition for oral cavity of the present invention is not particularly limited, for example, ointment, paste, pasta, gel, liquid, spray, mouthwash, liquid dentifrice, according to a conventional method. It can be in the form (form) of toothpaste, gum and the like. Among them, mouthwash, liquid dentifrice, toothpaste, ointment, paste, liquid and gel are preferable.

The oral composition of the present invention may further contain, alone or in combination, two or more optional components that can be incorporated into the oral composition, within a range that does not impair the effects of the present invention.

For example, as the surfactant, a nonionic surfactant, an anionic surfactant or an amphoteric surfactant can be blended. Specifically, for example, as nonionic surfactants, sugar fatty acid esters such as sucrose fatty acid ester, maltose fatty acid ester, lactose fatty acid ester; fatty acid alkanolamides; sorbitan fatty acid ester; fatty acid monoglyceride; polyoxyethylene addition coefficient is 8 -10, polyoxyethylene alkyl ether having an alkyl group having 13 to 15 carbon atoms; polyoxyethylene addition coefficient of 10 to 18 and polyoxyethylene alkylphenyl ether having an alkyl group having 9 carbon atoms; diethyl sebacate; Polyoxyethylene hydrogenated castor oil; fatty acid polyoxyethylene sorbitan and the like. As anionic surfactants, sulfate ester salts such as sodium lauryl sulfate and sodium polyoxyethylene lauryl ether sulfate; sulfosuccinate salts such as sodium lauryl sulfosulfosuccinate and sodium polyoxyethylene lauryl ether sulfosuccinate; cocoyl sarcosine sodium, lauroyl methylalanine Acyl amino acid salts such as sodium; cocoyl methyl taurine sodium and the like. As the zwitterionic surfactant, acetic acid betaine-type surfactants such as lauryldimethylaminoacetic acid betaine and coconut oil fatty acid amidopropyldimethylaminoacetic acid betaine; imidazoline-type such as N-cocoyl-N-carboxymethyl-N-hydroxyethylethylenediamine sodium Activators; amino acid type activators such as N-lauryldiaminoethylglycine and the like can be mentioned. These surfactants may be used alone or in combination of two or more. The blending amount is usually 0.1 to 5 mass% with respect to the total amount of the composition.

Further, as a flavoring agent, for example, menthol, carboxylic acid, anethole, eugenol, methyl salicylate, limonene, ocimene, n-decyl alcohol, citronell, 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, Fragrances such as d-borneol, fennel oil, cinnamon oil, cinnamaldehyde, peppermint oil, vanillin and the like can be used. These can be blended alone or in combination of two or more, for example, 0.001 to 1.5 mass% with respect to the total amount of the composition.

As the sweetener, for example, saccharin sodium, acesulfame potassium, stevioside, neohesperidyl dihydrochalcone, perillartine, thaumatin, asparatyl phenylalanyl methyl ester, p-methoxycinnamic aldehyde and the like can be used. These can be blended, for example, in an amount of 0.01 to 1% by mass based on the total amount of the composition.

Further, as a wetting agent, sorbit, ethylene glycol, propylene glycol, glycerin, 1,3-butylene glycol, polypropylene glycol, xylit, maltite, lactit, polyoxyethylene glycol, etc. may be blended alone or in combination of two or more. it can.

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

As the colorant, legal dyes such as Blue No. 1, Yellow No. 4, Red No. 202 and Green No. 3, mineral type dyes such as ultramarine blue, reinforced ultramarine blue and dark blue, titanium oxide and the like may be added.

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

The oral composition of the present invention includes not only cetylpyridinium chloride, but also vitamin Es such as dl-α-tocopherol acetate, tocopherol succinate, or tocopherol nicotinate as a medicinal component, and amphoteric amphoteric compounds such as dodecyldiaminoethylglycine. Disinfectant, triclosan, isopropylmethylphenol, nonionic disinfectant such as hinokitiol, anionic disinfectant such as sodium lauroylsarcosine, chlorhexidine hydrochloride, benzalkonium chloride, cationic disinfectant such as benzethonium chloride, dextranase, amylase , Enzymes such as protease, mutanase, lysozyme, lytic enzyme (Litech Enzyme), alkali metal monofluorophosphates such as sodium monofluorophosphate, potassium monofluorophosphate, fluorides such as sodium fluoride, stannous fluoride , Tranexamic acid, epsilon aminocaproic acid, aluminum chlorohydroxyl allantoin, dihydrocholesterol, glycyrrhetinic acid, glycyrrhizinic acid, copper chlorophyllin sodium, glycerophosphate, chlorophyll, sodium chloride, caropeptide, allantoin, carbazochrome, hinokitiol, potassium nitrate, palatinit, etc. alone Alternatively, two or more kinds may be combined and blended. These medicinal components may be used in combination with cetylpyridinium chloride, or may be added to the oral composition of the present invention containing no cetylpyridinium chloride.

It is also possible to add alcohols, silicon, apatite, white petrolatum, paraffin, liquid paraffin, microcrystalline wax, squalane, plastibase and the like as a base.

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

The term "comprising" includes "consisting essentially of" and "consisting of." in the present specification also includes "consisting essentially of" and "consisting of". Further, the invention includes all combinations of the constituent elements described in this specification.

In addition, the various characteristics (properties, structure, functions, etc.) described in the above embodiments of the present invention may be combined in any manner to identify the subject matter included in the present invention. That is, the invention includes all subject matter consisting of any combination of the combinable properties described herein.

Hereinafter, the present invention will be described more specifically, but the present invention is not limited to the following examples.

Examination of intercellular adhesion disruption inhibitory effect of retinol palmitate Gingival epithelial cell line Epi4 cells were cultured on a transwell until they became confluent. The transwell is a tool designed to hang the insert so that the insert is located in the middle of the well, and enables cell permeability evaluation.

In addition, a medium for cells containing 2.5% DMSO containing retinol palmitate was prepared. Retinol palmitate, which is a commercially available reagent, was used for the preparation, and the reagent contained 0.1% by mass, 0.05% by mass, 0.01% by mass, 0.005% by mass, or 0.001% by mass in the medium. It was made to be contained.

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

Furthermore, P. G. bacterium (P. gingivalis W83) was recovered by centrifugation at 10,000 rpm, and the concentration was adjusted so that OD600=0.5. Epi4 cells on the transwell after the above-mentioned incubation were added to P. 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-mentioned 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, if expressed 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 1IU=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 reagent is usually 0.9 to 0.95 mg/ml, about 118.25 mg/g×1/0.9 to 0.95=124 to 130 mg/ml, the palmitin The acid retinol reagent contains retinol palmitate. In addition, for example, in the study conducted by preparing a medium containing 0.1% by mass of the retinol palmitate reagent described above, the initial concentration was 900 to 950×118.25=106.4 to 112.3 μg/ml. The concentration is about 0.0106 to 0.0112 mass% in mass (w/w)% concentration. Similarly, in a study conducted by preparing a medium 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. It becomes 1/2, 1/10, 1/20, or 1/100 of 1 mass%. In addition, the final concentration (concentration after addition of P. g bacteria) is 3/4 of the concentration value.

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

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

Examination of intercellular adhesion disruption inhibitory effect of various vitamins Various vitamin compounds (specific compound names are shown in Fig. 3) Reagent (final concentration 0.0075%) was used, and concentration was adjusted so that OD600 = 0.1. Adjusted P. In the same manner as above, except that a culture solution of g. The results are shown in Figure 3. It was confirmed that among various vitamin compounds, retinol palmitate has a particularly excellent inhibitory effect on intercellular adhesion destruction.

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

WST-1, which is a cell growth activity reagent, 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 measuring the proliferative ability by analyzing the number of viable cells by the decomposition of the salt.

After the incubation, the absorbance (OD (450 nm)) was measured with an absorption plate reader (xMark microplate reader). The retinol palmitate concentration was converted so that the absorbance at 0 IU was 100%, and the cell viability upon treatment with each concentration of retinol palmitate was calculated. The results are shown in Fig. 4. No cytotoxicity was observed with retinol palmitate.

Examination of bactericidal activity of retinol palmitate P. g. bacterium (P. gingivalis W83) was cultured in a GAM medium to give O. It was adjusted to D(600)=1.0. The drug (retinol palmitate, cetylpyridinium chloride (CPC), or a combination thereof) adjusted to each concentration was mixed with the adjusted bacterial solution and left for 3 minutes. After 3 minutes, 10 times this amount of bactericide inactivating medium (0.07% lecithin + 0.5% Tween 80) was added to stop the bactericidal effect, and the cells were cultured at 37°C under anaerobic conditions for 24 hours. After culturing, the absorbance (OD (600)) of the medium was measured with a light absorption plate reader (xMark microplate reader), and the bactericidal effect was judged 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 for retinol palmitate, and no effect of inhibiting the bactericidal effect of CPC was observed.

Since retinol palmitate has no cytotoxicity or bactericidal activity, it was found in the study using the above transwell that the retinol palmitate lowered the FITC permeability. It was found that this was not because the cell-cell adhesion disruption was suppressed due to the g. g. strain being impaired, but because the cell-cell adhesion disruption was suppressed because the cell-cell adhesion of Epi4 cells was enhanced.

Claims (8)

An anti-periodontal disease oral composition containing retinol palmitate. The oral composition according to claim 1, which contains 0.0001 to 1% by mass of retinol palmitate. A composition for oral cavity containing 0.0001 to 1% by mass of retinol palmitate. The oral composition according to claim 1, which contains 1 to 10,000 IU of retinol palmitate. An oral composition containing 1 to 10,000 IU of retinol palmitate. The composition for oral cavity according to any one of claims 1 to 5, which is for enhancing adhesion between gingival epithelial cells. The oral composition according to claim 1, further comprising cetylpyridinium chloride. The oral composition according to claim 7, which contains 0.01 to 0.5% by mass of cetylpyridinium chloride.