JP2021031400A - Oral composition - Google Patents

Abstract

To efficiently inhibit gingipain produced by P. g bacteria with the fimA genotype being type II or IV.SOLUTION: The present invention relates to use of hinokitiol and/or copper chlorophyllin metal salt.SELECTED DRAWING: None

Description

The present disclosure relates to an oral composition or the like effective against Porphyromonas gingivalis (P. g bacterium) having a fimA genotype of type II. The contents of all documents described in this specification are incorporated herein by reference.

Porphyromonas gingivalis (P. g bacterium) is a periodontal disease bacterium having strong periodontal pathogenicity. P. Gingipine is known as a typical virulence factor produced by g-bacterium. Gingipine is a kind of protease, and Lys-zingipine (Kgp) and Arg-zingipine (Rgp) having different peptide cleavage site specificities exist. While interacting with each other, they cause disruption of connections between gingival epithelial cells, damage to the epithelial cells themselves and / or inhibition of proliferation, and thus inhibition of epithelial barrier destruction and repair. In addition, gingipine is said to have an effect of suppressing phagocytosis by phagocytic cells and intracellular digestion, as well as an effect of assisting the destruction of the complement system and the invasion into epithelial cells. It is also involved in avoiding bacteria g from the immune system. Furthermore, recently, it has been reported that the inhibitory effect of dysbiosis on the immune system leads to dysbiosis of the oral bacterial flora, and the action of these dysbiosis leads to the progression and intractability of periodontal disease.

In recent years, P.I. Attention has been paid to the gene fimA, which encodes fimbrillin, which is a ciliary constituent subunit of g fungus. There are 6 different genotypes (fimA type) of fimA due to the difference in nucleic acid sequence structure, and the severity of periodontal disease and P. gingivalis type II or IV fimA bacteria (particularly type II fimA bacteria) It has been suggested that a strong relevance is observed (Non-Patent Documents 1 and 2). Further, with respect to these six different genotypes (fimA type), the types can be easily confirmed by using, for example, PCR primers specific to each genotype (Non-Patent Document 2). Furthermore, it has been pointed out that there is a correlation between the strength of the fimA type Virulence and the genotype of gindipine (Non-Patent Document 3).

International Publication No. 2016/10524

"Porphyromonas gingivalis Relationship between cilia and periodontal pathogenicity" Japanese Periodontal Journal 45 (4): 357-363, 2003 "Distribution of periodontal disease-related bacterial species using genetic testing methods" Japan Clinical Microbiology Journal Vol. 28 No. 4 2018 Journal of Oral Microbiology 2015, 7: 29124

Therefore, P.I., whose fimA genotype is type II or type IV. It is considered that periodontal disease can be prevented and / or treated (particularly, periodontal disease progression is suppressed) particularly efficiently if the gingipine produced by g-bacterium can be efficiently suppressed.

By using hinokitiol and / or copper chlorophyllin metal salts, the present inventors used P.I. It was found that the gingipine produced by bacterium g can be suppressed particularly efficiently.

The present disclosure includes, for example, the subjects described in the following sections.
Item 1.
Contains hinokitiol and / or copper chlorophyllin metal salts,
For those who carry Porphyromonas gingivalis (P. g.) With fimA genotype II or IV in the oral cavity,
Anti-periodontal disease oral composition.
Item 2.
Contains hinokitiol and / or copper chlorophyllin metal salts,
For those who carry Porphyromonas gingivalis (P. g.) With fimA genotype II or IV in the oral cavity,
The P. g An oral composition for inhibiting a protease produced by a bacterium.
Item 3.
Item 2. The oral composition according to Item 1 or 2, which contains a hinokitiol and a copper chlorophyllin metal salt.
Item 4.
The fimA genotype is type II, P.I. Item 8. The oral composition according to any one of Items 1 to 3, for a person who has g-bacterium in the oral cavity.
Item 5.
P. held in the oral cavity. Among g bacteria, fimA genotype II type P. Item 8. The oral composition according to any one of Items 1 to 4, for a person having the highest amount of g-bacteria.

P. fimA genotype type II or type IV. Provided is an oral composition capable of efficiently suppressing gingipine produced by g-bacterium.

Due to various bactericidal and anti-inflammatory components, P.I. The result of examination whether it is possible to suppress the gindipine activity of g bacterium (type IV fimA strain) is shown. Due to various bactericidal and anti-inflammatory components, P.I. The result of examination whether it is possible to suppress the gindipine activity of g bacterium (type IV fimA strain) is shown. With sodium copper chlorophyllin, P. The results of examining whether it is possible to suppress the gingipine activity produced by each strain of g bacterium (type II fimA strain or type IV fimA strain) are shown. By hinokitiol, P. The results of examining whether it is possible to suppress the gingipine activity produced by each strain of g bacterium (type II fimA strain or type IV fimA strain) are shown. When copper chlorophyllin sodium and hinokitiol were used in combination, P.I. The results of examining the effect of suppressing the gingipain activity of bacterium g (type II fimA strain) are shown. When copper chlorophyllin sodium and isopropylmethylphenol were used in combination, P.I. The results of examining the effect of suppressing the gingipain activity of bacterium g (type II fimA strain) are shown. When copper chlorophyllin sodium and chlorohexidine hydrochloride were used in combination, P.I. The results of examining the effect of suppressing the gingipain activity of bacterium g (type II fimA strain) are shown. When copper chlorophyllin sodium and cetylpyridinium chloride were used in combination, P.I. The results of examining the effect of suppressing the gingipain activity of bacterium g (type II fimA strain) are shown.

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

The oral compositions included in the present disclosure contain hinokitiol and / or copper chlorophyllin metal salts. As the copper chlorophyllin metal salt, a copper chlorophyllin alkali metal salt is preferable, and among them, copper chlorophyllin sodium or copper chlorophyllin potassium is preferable. The oral composition may be referred to as the oral composition of the present disclosure.

The oral composition of the present disclosure is preferably used as an anti-periodontal oral composition for a person who has Porphyromonas gingivalis (P. g bacterium) having a fimA genotype of type II or IV in the oral cavity. Can be done. More specifically, it can be preferably used, for example, for the prevention and / or treatment of periodontal disease for the person concerned, or for suppressing the progression of periodontal disease.

Further, as described above, in the oral composition of the present disclosure, the fimA genotype is P.I. Since the gingipine produced by the g bacterium can be efficiently suppressed, the P. It can also be said to be an oral composition for inhibiting protease produced by g bacteria.

In addition, P. There are a total of 6 types of fimA genotypes of bacterium g, type I, type Ib, type II, type III, type IV, and type V, and P.I. There are also people who carry g bacteria. Of these, type II is considered to be the most pathogenic. Therefore, the oral composition of the present disclosure is particularly contained in the oral cavity. Among g bacteria, fimA genotype II type P. It can be preferably used for those who have the most g-bacteria.

Among the oral compositions, for example, an oral composition containing hinokitiol and a copper chlorophyllin metal salt is particularly preferable. The combination of hinokitiol and copper chlorophyllin metal salts is particularly excellent, and the fimA genotype is type II or type IV. This is because it exerts the effect of suppressing gingipine produced by bacterium g.

The amount of hinokitiol contained in the oral composition is preferably 0.003% by mass or more, for example. The upper limit is not particularly limited, but for example, 0.2% by mass can be exemplified. The amount range is preferably, for example, about 0.003 to 0.2% by mass. The upper or lower limit of the range is, for example, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.015, 0.02, 0.025, 0. .03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09 , 0.095, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, or 0.19% by mass. May be good. For example, it may be about 0.004 to 0.15% by mass or 0.005 to 0.1% by mass.

The amount of copper chlorophyllin metal salt contained in the oral composition is preferably 0.003% by mass or more, for example. The upper limit is not particularly limited, but for example, 0.2% by mass can be exemplified. The amount range is preferably, for example, about 0.003 to 0.2% by mass. The upper or lower limit of the range is, for example, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.015, 0.02, 0.025, 0. .03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09 , 0.095, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, or 0.19% by mass. May be good. For example, it may be about 0.004 to 0.15% by mass or 0.005 to 0.1% by mass.

The oral composition can be a solid composition or a liquid composition. The oral composition can be used, for example, as a pharmaceutical product or a quasi-drug. The form is not particularly limited, but according to a conventional method, for example, an ointment, a paste, a pasta, a gel, a liquid, a spray, a mouthwash, a liquid dentifrice, a dentifrice, a gum, etc. It can be in the form (dosage form). Of these, mouthwashes, liquid dentifrices, dentifrices, ointments, pastes, liquids and gels are preferable.

The oral composition may contain any component that can be blended in the oral composition alone or in combination of two or more, as long as the above effects are not impaired.

For example, as a surfactant, a nonionic surfactant, an anionic surfactant or an amphoteric surfactant can be blended. Specifically, for example, sugar fatty acid esters such as sucrose fatty acid ester, maltose fatty acid ester, and lactose fatty acid ester; fatty acid alkanolamides; sorbitan fatty acid ester; fatty acid monoglyceride; polyoxyethylene addition coefficient is 8 as a nonionic surfactant. A polyoxyethylene alkyl ether having a carbon number of 10 to 10 and an alkyl group having 13 to 15; a polyoxyethylene alkylphenyl ether having a polyoxyethylene addition coefficient of 10 to 18 and an alkyl group having a carbon number of 9; diethyl sebacate; Polyoxyethylene hydrogenated castor oil; fatty acid polyoxyethylene sorbitan and the like can be mentioned. Examples of the anionic surfactant include sulfate esters 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 sarcosin and lauroyl methyl alanine. Acylamino acid salts such as sodium; cocoyl methyl taurine sodium and the like can be mentioned. Examples of amphoteric ion surfactants include betaine acetate-type surfactants such as lauryldimethylaminoacetate betaine and coconut oil fatty acid amide propyldimethylaminoacetic acid betaine; and imidazoline-type activators such as N-cocoyl-N-carboxymethyl-N-hydroxyethylethylenediamine sodium. Activator: Amino acid type activator such as N-lauryldiaminoethylglycine and the like can be mentioned. These surfactants can be blended alone or in combination of two or more. The blending amount is usually 0.1 to 5% by mass based on the total amount of the composition.

Also, as flavoring agents, for example, menthol, carboxylic acid, anator, eugenol, methyl salicylate, limonene, osimene, n-decyl alcohol, citronell, α-terpineol, methylacetate, citronenylacetate, methyleugenol, cineol, etc. Linalool, ethyl linalool, timol, spearmint oil, peppermint oil, lemon oil, orange oil, sage oil, rosemary oil, siliceous oil, perilla oil, winter green oil, clove oil, eucalyptus oil, piment oil, d-campul, Fragrances such as d-borneol, orange oil, kehi oil, limonene aldehyde, peppermint oil, and vanillin can be used. These can be blended, for example, 0.001 to 1.5% by mass with respect to the total amount of the composition, either alone or in combination of two or more.

Further, as the sweetener, for example, sodium saccharin, potassium acesulfarm, stebioside, neohesperidyl dihydrochalcone, perillartine, taumatin, aspalathylphenylalanyl methyl ester, p-methoxycinnamic aldehyde and the like can be used. These can be blended, for example, 0.01 to 1% by mass with respect to the total amount of the composition.

Further, as a wetting agent, sorbitol, ethylene glycol, propylene glycol, glycerin, 1,3-butylene glycol, polypropylene glycol, xylit, martit, lacchit, polyoxyethylene glycol and the like may be blended alone or in combination of two or more. it can.

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

As the colorant, legal pigments such as Blue No. 1, Yellow No. 4, Red No. 202, and Green No. 3, mineral pigments such as ultramarine, enhanced ultramarine, and dark blue, titanium oxide, and the like may be blended.

As the pH adjuster, citric acid, phosphoric acid, malic acid, pyrophosphate, lactic acid, tartaric acid, glycerophosphate, acetic acid, nitric acid, chemically possible salts thereof, sodium hydroxide and the like may be blended. 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 blending amount of the pH adjuster may be, for example, 0.01 to 2% by weight.

To the extent that the effect is not impaired, the oral composition further contains vitamin Es such as dl-α-tocopherol acetate, tocopherol succinate, or tocopherol nicotinate, and cations such as cetylpyridinium chloride as medicinal ingredients. Sex bactericides, anionic bactericides such as sodium lauroyl sarcosin, amphoteric bactericides such as dodecyldiaminoethylglycine, nonionic bactericides such as triclosan and isopropylmethylphenol, dextranase, amylase, protease, mutanase, lysozyme, lytic bacterium Enzymes such as enzymes (Litec Enzyme), alkali metal monofluorophosphates such as sodium monofluorophosphate and potassium monofluorophosphate, fluorides such as sodium fluoride and stannous fluoride, tranexamic acid and epsilon aminocaproic acid, Aluminum chlorhydroxylphenolin, dihydrocholesterol, glycyrrhetinic acid, glycyrrhizic acid, glycerofluoride, chlorophyll, sodium chloride, caropeptide, allantin, carbazochrome, hinokithiol, potassium nitrate, palatinit, etc. can be blended alone or in combination of two or more. ..

Further, as a base, alcohols, silicon, apatite, white petrolatum, paraffin, liquid paraffin, microcrystalline wax, squalane, plastibase and the like can be added.

In addition, the oral composition can be prepared by a known method or a method easily conceived from a known method. For example, it can be prepared by appropriately mixing hinokitiol and / or copper chlorophyllin metal salt and, if necessary, other components.

In addition, in this specification, "including" also includes "consisting essentially" and "consisting of" (The term "comprising" includes "consisting essentially of" and "consisting of."). The present disclosure also includes all combinations of the constituent requirements described herein.

In addition, the various properties (property, structure, function, etc.) described for each embodiment of the present disclosure described above may be combined in any way in specifying the subject matter included in the present disclosure. That is, the present disclosure includes all subjects consisting of any combination of each combinable property described herein.

Hereinafter, embodiments of the present disclosure will be described in more detail with reference to examples, but the embodiments of the present disclosure are not limited to the following examples. The amounts of the various components to be examined may be indicated by the concentration (ppm) below, and although the mass per 1 ml of the liquid used is slightly larger than 1 g, it can be converted to about 1000 ppm = 0.1 mass%.

Search for Gingipine activity-suppressing components The effects of various bactericidal and anti-inflammatory components on Gingipine activity-suppressing components were investigated. Ingredients to be examined include sodium copper chlorophyllin, hinokithiol, isopropylmethylphenol (IPMP), cetylpyridinium chloride (CPC), chlorohexidine hydrochloride (CHX), dipotassium glycyrrhizinate (GK2), β-glycyrrhetinic acid, tranexamic acid, allantin, aminocaproic acid. , Vitamin B6 (VB6), and Vitamin E Acetate (VEA; also referred to as dl-α-tocopherol acetate). In addition, P. As the g-bacterium, W83, which is a type IV fimA strain, was used.

P. Bacteria g (P. gingivalis W83) was cultured in GAM medium, and the absorbance (OD (600)) was adjusted to 1.0. The bacterial solution was centrifuged at 10000 rpm, and the supernatant was collected. Gingipine is contained in the collected supernatant. 200 μL of a solution prepared by adjusting various study components to each concentration by 10% DMSO was mixed with 20 μL of the supernatant and left for 3 minutes. After 3 minutes, it was mixed with a substrate of gingipine (Z-Phe-Arg-MCA; Peptide Institute, Ltd.) diluted 100-fold with PBS, and allowed to stand at 37 ° C. for 1 hour in the dark. Z-Phe-Arg-MCA is Benzyloxycarbonyl-L-phenylalanyl-L-arginine 4-methylcoumaryl-7-amide (Hydrochloride Form), which is a reagent that is cleaved by Arg-zingipine (Rgp) activity to emit fluorescence. After 1 hour, the fluorescence intensity (excitation light: 380 nm, emitted light: 440 nm) was measured with a fluorescence plate reader (Gemini XPS). The fluorescence intensity at a concentration of the study component of 0 μg / mL was converted to 100%, and the gingipine activity when the study component of each concentration was treated was calculated. The examination was carried out with n = 2 for each concentration, and the calculated value was an average value. The smaller the calculated value, the more the gingipine activity was suppressed. The results are shown in FIGS. 1a and 1b.

Of the various components used in the study, only hinokitiol and sodium copper chlorophyllin were found to have an inhibitory effect on gindipine activity.

Examination of the inhibitory effect of hinokitiol and sodium copper chlorophyllin on gindipine activity P. In addition to W83, OMZ314, which is a type II fimA strain, and W50, which is a type IV fimA strain, are used as strains of g bacteria, and Z-His-Glu-not only Z-Phe-Arg-MCA but also Z-His-Glu- is used as a substrate for gingipine. Using Lys-MCA (Peptide Institute, Ltd.), the inhibitory effect of hinokitiol and sodium copper chlorophyllin on gindipine activity was examined in the same manner as described above. Z-His-Glu-Lys-MCA is Benzyloxycarbonyl-L-Histidyl-L-Glutamyl-L-Lysine 4-methylcoumaryl-7-amide (Hydrochloride Form), which is cleaved by Lys-zingipine (Kgp) activity. It is a reagent that emits fluorescence.

The results using sodium copper chlorophyllin are shown in FIG. 2a, and the results using hinokitiol are shown in FIG. 2b. In these figures, the horizontal axis indicates the concentration (ppm) of sodium copper chlorophyllin or hinokitiol.

Examination of the effect of suppressing gingipine activity by a combination of bactericidal components P.I. The activity-suppressing effect of gingipine produced by bacterium g (OMZ314, which is a type II fimA strain) was examined. Specifically, the effect of using sodium copper chlorophyllin in combination with hinokitiol, isopropylmethylphenol (IPMP), chlorohexidine hydrochloride (CHX), or cetylpyridinium chloride (CPC) was examined in the same manner as above. did. The results of the combination of sodium copper chlorophyllin and hinokithiol are shown in FIG. 3a, the results of the combination of sodium copper chlorophyllin and IPMP are shown in FIG. 3b, and the results of the combination of sodium copper chlorophyllin and CHX hydrochloride are shown in FIG. 3c. The results of the combination of sodium copper chlorophyllin and CPC are shown in FIG. 3d, respectively. In these figures, the vertical axis represents gindipine (Rgp) activity (%). The concentration of each bactericidal component is shown in% by mass. In addition, "copper black" indicates sodium copper chlorophyllin.

Examination of bactericidal components other than hinokithiol Even when used in combination with sodium copper chlorophyllin, it shows the same effect of suppressing gingipine activity as when sodium copper chlorophyllin is used alone, or rather than when sodium copper chlorophyllin is used alone. However, the effect of suppressing gingipane activity was reduced. On the other hand, when copper chlorophyllin sodium and hinokitiol were used in combination, a particularly excellent effect of suppressing gingipane activity was obtained.

Claims (5)

Contains hinokitiol and / or copper chlorophyllin metal salts,
For those who carry Porphyromonas gingivalis (P. g.) With fimA genotype II or IV in the oral cavity,
Anti-periodontal disease oral composition. Contains hinokitiol and / or copper chlorophyllin metal salts,
For those who carry Porphyromonas gingivalis (P. g.) With fimA genotype II or IV in the oral cavity,
The P. g An oral composition for inhibiting a protease produced by a bacterium. The oral composition according to claim 1 or 2, which contains a hinokitiol and a copper chlorophyllin metal salt. The fimA genotype is type II, P.I. The oral composition according to any one of claims 1 to 3, for a person who has g bacteria in the oral cavity. P. held in the oral cavity. Among g bacteria, fimA genotype II type P. The oral composition according to any one of claims 1 to 4, for a person having the highest amount of g-bacterium.