Composition containing hinokitiol and zinc gluconate

Abstract

To provide a composition that inhibits neutrophil collagenase activity.SOLUTION: An oral composition contains hinokitiol and zinc gluconate. A content of the zinc gluconate is 2-150 pts.mass relative to 1 pt.mass of the hinokitiol.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present disclosure relates to a composition containing hinokitiol and zinc gluconate, and uses thereof. [Background technology]

[0002] Matrix metalloproteinases (MMPs) are a group of enzymes that contain a zinc(II) ion at their active site and degrade extracellular matrix metalloproteinases (EMMs). Their main substrates are biopolymers such as collagen, laminin, gelatin, and fibronectin.

[0003] MMPs are thought to maintain homeostasis by degrading unnecessary extracellular matrix and controlling angiogenesis and the construction of new tissues during tissue remodeling and wound healing in vivo. However, excessive MMP activity can lead to the progression of tissue destruction and is thought to be involved in periodontal disease, rheumatoid arthritis, tumor invasion and metastasis, among other phenomena. Therefore, compounds that can regulate MMP activity are being sought. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 5339708 [Non-patent literature]

[0005] [Non-Patent Document 1] Japanese Journal of Periodontology 59(4):185-190,2017 [Non-patent document 2] J Clin Periodontol.2021 Aug;48(8):1051-1065. Summary of the Invention [Problem to be solved by the invention]

[0006] MMP-8, also known as neutrophil collagenase because it is a collagenase produced by neutrophils, is known to primarily degrade extracellular matrix such as collagen and accounts for approximately 80% of the MMP in gingival crevicular fluid. Neutrophils accumulate in gingival crevicular fluid in response to inflammation in the periodontal pocket and produce neutrophil collagenase (MMP-8), which is thought to destroy the junctional epithelium in the periodontal pocket, deepen the periodontal pocket, and progress periodontal disease (Non-Patent Document 1). Therefore, there is a need to identify components that can control the activity of neutrophil collagenase in order to prevent the progression of periodontal disease (Patent Document 1).

[0007] An object of the present disclosure is to provide a composition that inhibits neutrophil collagenase activity. Another object is to provide a composition for preventing the progression of periodontal disease. [Means for solving the problem]

[0008] The present inventors have found that the use of hinokitiol and zinc gluconate in a specific ratio exhibits excellent inhibitory activity against neutrophil collagenase, and have made further improvements.

[0009] The present disclosure encompasses, for example, the subject matter described in the following sections: Section 1. Contains hinokitiol and zinc gluconate, An oral composition, comprising 2 to 150 parts by mass of zinc gluconate per part by mass of hinokitiol. Section 2. Item 1. The composition according to item 1 for preventing the progression of periodontal disease. Section 3. Item 3. The composition according to Item 1 or 2, which is used for at least one purpose selected from the group consisting of inhibiting neutrophil collagenase activity, suppressing gingival recession, suppressing periodontal pocket formation, suppressing periodontal pocket deepening, suppressing periodontal tissue destruction, suppressing gum recession, and suppressing gingival collagen degradation. Section 4. Item 4. The composition according to any one of Items 1 to 3, wherein the content of hinokitiol in the composition is 0.0005% by mass or more. Section 5. Item 5. The composition according to any one of Items 1 to 4, wherein the content of zinc gluconate in the composition is 0.001% by mass or more. [Effects of the Invention]

[0010] Neutrophil collagenase activity can be inhibited, and an effective composition for preventing the progression of periodontal disease can be provided. [Brief explanation of the drawings]

[0011] [Figure 1] 1 shows the results of measuring neutrophil collagenase activity when hinokitiol and zinc gluconate were added. DETAILED DESCRIPTION OF THE INVENTION

[0012] Each embodiment included in the present disclosure will be described in further detail below. The composition encompassed by the present disclosure contains hinokitiol and zinc gluconate. In this specification, the composition may be referred to as the "composition of the present disclosure."

[0013] Hinokitiol has the following formula:

[0014] [ka]

[0015] Hinokitiol used in the composition of the present disclosure may be a synthetic product or may be extracted from a natural product (such as Japanese cypress).

[0016] The content of hinokitiol in the composition of the present disclosure is not particularly limited as long as the effect is achieved. For example, the content of hinokitiol in the composition of the present disclosure can be 0.0005% by mass or more. The upper or lower limit of this range may be, for example, 0.0006, 0.0007, 0.0008, 0.0009, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, or 0.3% by mass. For example, it may be 0.0005 to 0.3% by mass or 0.001 to 0.3% by mass.

[0017] The content of zinc gluconate in the composition of the present disclosure is not particularly limited as long as it is within a range that exhibits the desired effect. For example, the content of zinc gluconate in the composition of the present disclosure can be 0.01% by mass or more. The upper or lower limit of this range may be, for example, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, or 0.5% by mass. For example, it may be 0.01 to 0.5% by mass, or 0.05 to 0.3% by mass.

[0018] The content of zinc gluconate per 1 part by mass of hinokitiol in the composition of the present disclosure is preferably 2 to 150 parts by mass. The upper or lower limit of this range may be, for example, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, or 125 parts by mass. For example, it may be 3 to 125 parts by mass or 3 to 100 parts by mass.

[0019] Neutrophil collagenase (EC 3.4.24.34) is a collagenase produced by neutrophils and is also called MMP-8.

[0020] Inhibition of neutrophil collagenase activity means inhibiting the action of neutrophil collagenase to decompose a substrate (for example, collagen such as type I collagen, type II collagen, and type III collagen).

[0021] The composition of the present disclosure inhibits the degradation of substrates by neutrophil collagenase, and therefore can suppress the destruction of tissues (e.g., junctional epithelium, gingival sulcus epithelium, periodontal ligament, etc.) by neutrophil collagenase, the degradation of gingival collagen (collagen fibers that constitute the gingiva), etc. Furthermore, the composition of the present disclosure has an inhibitory effect on neutrophil collagenase activity, and therefore can inhibit the activity of neutrophil collagenase that has already been produced.

[0022] For this reason, the composition of the present disclosure is suitable as an oral composition. More specifically, the composition of the present disclosure can be used for preventing the progression of periodontal disease, such as inhibiting the progression of periodontal disease, inhibiting neutrophil collagenase activity, inhibiting gingival recession, inhibiting periodontal pocket formation, inhibiting periodontal pocket deepening, inhibiting periodontal tissue destruction, inhibiting gingival recession, inhibiting gingival collagen degradation, inhibiting attachment loss, protecting periodontal tissue collagen, and inhibiting periodontal tissue destruction associated with chronic inflammation, and / or for protecting dentin collagen, etc. As used herein, "inhibition of periodontal pocket formation" means inhibiting the formation of periodontal pockets (grooves between the teeth and gums). As used herein, "inhibition of periodontal pocket deepening" means inhibiting the deepening of periodontal pockets (grooves between the teeth and gums). As used herein, "inhibition of attachment loss" means inhibiting the gingival junctional epithelium from peeling off from the tooth surface, causing the attachment position between the gums and the teeth to shift toward the tooth root. As used herein, "inhibition of gingival recession" means inhibiting the entire gums from moving toward the tooth root. As used herein, "inhibition of gingival recession" means inhibiting the exposure of the tooth root due to the entire gums moving toward the tooth root.

[0023] The composition of the present disclosure is preferably applied to subjects with elevated neutrophil collagenase activity. More specifically, examples include individuals who fall into stage 1, 2, 3, or 4, or grade C, of ​​the diagnostic criteria for periodontal disease (Non-Patent Document 2). The diagnostic criteria for periodontal disease were developed at a workshop cosponsored by the American Academy of Periodontology and the European Federation of Periodontology, and can be found on the Japanese Society of Periodontology website (https: / / www.perio.jp / file / news / info_191220.pdf). Examples of individuals who fall into stage 1 of the diagnostic criteria for periodontal disease include those with a maximum interdental clinical attachment level (the distance from the cement-enamel junction to the gingival sulcus epithelium (measured in 1 mm increments)) of approximately 1 to 2 mm. Examples of individuals who fall into stage 2 of the diagnostic criteria for periodontal disease include those with a maximum interdental clinical attachment level of approximately 3 to 4 mm. Examples of people who fall into stage 3 or 4 according to the diagnostic criteria for periodontal disease include people whose interdental clinical attachment level is 5 mm or greater. Among these, examples of people who fall into stage 3 according to the diagnostic criteria for periodontal disease include people who have lost 1 to 4 teeth due to periodontal disease. Examples of people who fall into stage 4 according to the diagnostic criteria for periodontal disease include people who have lost 5 or more teeth due to periodontal disease. Examples of people who fall into grade C according to the diagnostic criteria for periodontal disease include people whose clinical attachment level has changed by 2 mm or greater over 5 years. Furthermore, examples of subjects to which the composition of the present disclosure can be applied include mammals including humans (for example, dogs, cats, mice, rats, sheep, horses, cows, monkeys, etc.), with humans being preferred.

[0024] The composition of the present disclosure may be, for example, a solid composition, a liquid composition, etc. Furthermore, the composition of the present disclosure (particularly an oral composition) can be made into the form (dosage form) of, for example, an ointment, a paste, a paste, a gel, a liquid, a spray, a mouthwash, a liquid dentifrice, a toothpaste, a gum, a tablet, a drop, etc. in accordance with a conventional method. Among these, a mouthwash, a liquid dentifrice, a toothpaste, an ointment, a paste, a liquid, or a gel is preferred.

[0025] The composition of the present disclosure may further contain, for example, one or more optional components that can be incorporated into oral compositions, as long as the effects are not impaired.

[0026] For example, nonionic surfactants, anionic surfactants, or amphoteric surfactants can be blended as surfactants. Specific examples of 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; glycerin fatty acid esters; sorbitan fatty acid esters; fatty acid monoglycerides; polyoxyethylene alkyl ethers having a polyoxyethylene addition coefficient of 8 to 10 and an alkyl group having 13 to 15 carbon atoms; polyoxyethylene alkylphenyl ethers having a polyoxyethylene addition coefficient of 10 to 18 and an alkyl group having 9 carbon atoms; diethyl sebacate; polyoxyethylene hydrogenated castor oil; and fatty acid polyoxyethylene sorbitan. Examples of anionic surfactants include sulfate ester salts such as sodium lauryl sulfate and polyoxyethylene lauryl ether sodium sulfate; sulfosuccinates such as sodium lauryl sulfosuccinate and polyoxyethylene lauryl ether sodium sulfosuccinate; acylamino acid salts such as sodium cocoyl sarcosinate and sodium lauroyl methyl alanine; and sodium cocoyl methyl taurate. Examples of zwitterionic surfactants include acetate betaine surfactants such as lauryl dimethylaminoacetate betaine and coconut oil fatty acid amidopropyl dimethylaminoacetate betaine; and imidazoline surfactants such as N-cocoyl-N-carboxymethyl-N-hydroxyethylethylenediamine sodium. These surfactants can be used alone or in combination of two or more. The amount of surfactant used is usually 0.1 to 5% by mass based on the total amount of the composition.

[0027] Flavoring agents that can be used include, for example, menthol, carvone, anethole, eugenol, methyl salicylate, limonene, ocimene, n-decyl alcohol, citronellol, α-terpineol, methyl acetate, citronenyl acetate, methyl eugenol, cineole, 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, d-borneol, fennel oil, cinnamon oil, cinnamon aldehyde, peppermint oil, vanillin, etc. These can be blended alone or in combination of two or more types in an amount of, for example, 0.001 to 1.5% by mass based on the total amount of the composition.

[0028] In addition, sweeteners that can be used include, for example, saccharin sodium, acesulfame potassium, stevioside, neohesperidyl dihydrochalcone, perillartine, thaumatin, aspartyl phenylalanyl methyl ester, p-methoxycinnamic aldehyde, etc. These can be blended in an amount of, for example, 0.01 to 1% by mass based on the total amount of the composition.

[0029] Furthermore, sorbitol, ethylene glycol, propylene glycol, glycerin, 1,3-butylene glycol, polypropylene glycol, xylitol, maltitol, lactite, polyoxyethylene glycol, etc. may be blended alone or in combination of two or more kinds as a humectant.

[0030] Examples of binders include cellulose derivatives such as sodium carboxymethylcellulose, carboxymethylethylcellulose salts, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose, ethylcellulose, crystalline cellulose, and crystalline cellulose-carmellose sodium; microbial polymers such as xanthan gum; natural polymers or natural rubbers such as tragacanth gum, karaya gum, gum arabic, carrageenan, dextrin, agar, pectin, pullulan, gellan gum, locust bean gum, and sodium alginate; synthetic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, polyvinyl methyl ether, and sodium polyacrylate; thickening silica; inorganic binders such as Veegum; and cationic binders such as O-[2-hydroxy-3-(trimethylammonio)propyl]hydroxyethylcellulose chloride. These binders can be used alone or in combination.

[0031] As preservatives, parabens such as methylparaben, ethylparaben, propylparaben, and butylparaben, sodium benzoate, phenoxyethanol, alkyldiaminoethylglycine hydrochloride, and the like can be added.

[0032] As coloring agents, 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 Prussian blue, titanium oxide, and the like may be blended.

[0033] The pH adjuster may be citric acid, phosphoric acid, malic acid, pyrophosphoric acid, lactic acid, tartaric acid, glycerophosphoric acid, acetic acid, nitric acid, or chemically acceptable salts thereof, or sodium hydroxide. These may be added 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 amount of pH adjuster added may be, for example, 0.01 to 2% by weight.

[0034] The oral composition of the present disclosure may further contain, as active ingredients, vitamin E compounds such as dl-α-tocopherol acetate, tocopherol succinate, or tocopherol nicotinate; amphoteric bactericides such as dodecyldiaminoethylglycine; nonionic bactericides such as triclosan and isopropylmethylphenol; anionic bactericides such as sodium lauroyl sarcosinate; cationic bactericides such as cetylpyridinium chloride, chlorhexidine hydrochloride, benzalkonium chloride, and benzethonium chloride; dextranase, amylase, protease, mutanase, lysozyme, and lytic enzymes (lytic enzymes). The following may be blended alone or in combination of two or more: enzymes such as sodium monofluorophosphate, potassium monofluorophosphate, and other alkali metal monofluorophosphates; fluorides such as sodium fluoride and stannous fluoride; tranexamic acid, epsilon aminocaproic acid, aluminum chlorohydroxyl allantoin, dihydrocholesterol, glycyrrhetinic acid, glycyrrhizinic acid, copper chlorophyllin sodium, glycerophosphate, chlorophyll, sodium chloride, callopeptide, allantoin, carbazochrome, potassium nitrate, palatinit, and the like.

[0035] Furthermore, it is also possible to add alcohols, silicone, apatite, white petrolatum, paraffin, liquid paraffin, microcrystalline wax, squalane, plastibase, etc. as bases.

[0036] It should be noted that in this specification, the term "comprising" includes "consisting essentially of" and "consisting of." In addition, the present disclosure encompasses all arbitrary combinations of the constituent elements described in this specification.

[0037] Furthermore, the various characteristics (properties, structures, functions, etc.) described in each embodiment of the present disclosure above may be combined in any way to specify the subject matter encompassed by the present disclosure, i.e., the present disclosure encompasses all subject matter consisting of any combination of the combinable characteristics described herein. [Example]

[0038] The contents of the present disclosure will be specifically explained using the following experimental examples. However, the present disclosure is not limited to these in any way. In the following, unless otherwise specified, experiments were performed under atmospheric pressure and room temperature conditions. Furthermore, unless otherwise specified, "%" means "(weight / volume)%."

[0039] Inhibition of neutrophil collagenase activity 1. Hinokitiol and zinc gluconate were dissolved in the solvent (buffer provided with the kit + 5% DMSO) to a concentration twice the desired addition concentration. Equal amounts of hinokitiol and zinc gluconate of each concentration were mixed to prepare a mixed solution of the desired concentration. 2. Neutrophil collagenase activity was evaluated using the MMP-8 fluorimetric drug discovery kit (Enzo Life Sciences, Inc., BML-AK415-0001). Specifically, 20 μl of each material solution prepared in step 1 above, 20 μl of 200-fold diluted MMP-8, and 50 μl of the buffer provided with the kit were mixed and incubated at 37°C for 60 minutes. A positive control sample was prepared by adding an equal volume of buffer without adding any material, and a sample was prepared by adding an equal volume of buffer without adding MMP-8 to measure the background signal of the substrate alone. 3. The fluorescent substrate (BML-P126-9090) was dissolved and diluted 10-fold with buffer (40 μM concentration). 10 μl of the diluted fluorescent substrate was added, and after 20 minutes of reaction at 37°C, the fluorescence intensity was measured at excitation light of 328 nm and emission light of 420 nm. The fluorescence intensity of the positive control sample minus the fluorescence intensity of the substrate alone as background (NC) was set at 100%, and the enzyme activity was calculated as the ratio of the fluorescence intensity of each sample with added material solution minus the fluorescence intensity of the substrate alone as background.

[0040] To evaluate the effect of the combined use of hinokitiol and zinc gluconate, the Combination Index (CI) value was calculated according to the following formula (TING-CHAO CHOU, et al. Trends Pharmacol Sci. 1983;4:450-454, C Patrick Reynolds, et al. Methods Mol Med. 2005;110:173-83.). CI=(D)1 / (Dx)1+(D)2 / (Dx)2 (D) 1: Hinokitiol addition concentration (D) 2: Zinc gluconate addition concentration (Dx) 1: The concentration of hinokitiol alone that inhibits neutrophil collagenase activity by 50% (IC50 value) (Dx) 2: The concentration of zinc gluconate alone that inhibits neutrophil collagenase activity by 50% (IC50 value) When CI<1, it was judged that there was a synergistic effect, when CI=1, there was an additive effect, and when CI>1, there was an antagonistic effect. The results are shown in Table 1. The concentrations in Table 1 indicate the concentrations of each material when adjusted in 1 above, which are five times higher than the concentrations in the reaction system in 3 above. This is because when oral compositions such as toothpaste are used, they are diluted by saliva, etc., so the evaluation was conducted under the assumption that they would actually be used in the oral cavity.

[0041] [Table 1]

[0042] As shown in Table 1, a synergistic effect was observed in the inhibitory activity against neutrophil collagenase when the zinc gluconate:hinokitiol ratio was between 100:1 and 3.125:1. Furthermore, in the combinations where a synergistic effect was confirmed (100:1, 50:1, 25:1, 12.5:1, 6.25:1, and 3.125:1), neutrophil collagenase activity was confirmed to be reduced by approximately 45, 42, 41, 44, 39, and 49%, respectively. Some of the results are shown in Figure 1. The concentrations in Figure 1 indicate the concentrations of each material at the time of preparation in step 1 above.

Claims

1. Contains hinokitiol and zinc gluconate, An oral composition comprising 2 to 150 parts by mass of zinc gluconate per part by mass of hinokitiol.

2. The composition according to claim 1 for preventing the progression of periodontal disease.

3. The composition according to claim 1 or 2, which is for use in at least one application selected from the group consisting of inhibiting neutrophil collagenase activity, inhibiting gingival recession, inhibiting periodontal pocket formation, inhibiting periodontal pocket deepening, inhibiting periodontal tissue destruction, inhibiting gum recession, and inhibiting gingival collagen degradation.

4. The composition according to any one of claims 1 to 3, wherein the content of hinokitiol in the composition is 0.0005% by mass or more.

5. The composition according to any one of claims 1 to 4, wherein the content of zinc gluconate in the composition is 0.01% by mass or more.

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