JP2021031406A - Gingival tissue destruction inhibitor - Google Patents

Abstract

To provide oral compositions used for inhibiting the destruction of gingival tissue by collagen degrading enzymes, also to provide oral compositions used for inhibiting or improving gingival recession caused by the destruction of gingival tissue by collagen degrading enzymes, and additionally to provide oral compositions used for inhibiting the production of collagen-degrading enzymes due to bacterial infections in the periodontal tissue.SOLUTION: Hinokitiol is contained as an active ingredient to prepare a gingival tissue destruction inhibitor, a gingival recession inhibitor or improver, a gingival barrier function enhancer, or an inhibitor of collagen-degrading enzyme production in periodontal tissue.SELECTED DRAWING: None

Description

The present invention relates to an oral composition used to suppress the destruction of gingival tissue by the collagen degrading enzyme MMP-1. The present invention also relates to an oral composition used for suppressing destruction of gingival tissue and suppressing or ameliorating gingival retraction. Furthermore, the present invention relates to an oral composition used to suppress the expression and production of MMP-1 in periodontal tissues.

Periodontal disease is an inflammatory disease of the periodontal tissue, caused by the host response to bacterial invasion and infection. In particular, Porphyromonas gingivalis is a bacterium that is considered to be the most common pathogen of adult periodontal disease among periodontal diseases.

The gingival (gingival) tissue is a periodontal tissue that supports the teeth, and is composed of gingival cells (gingival epithelial cells, gingival fibroblasts, etc.) and an extracellular matrix such as collagen produced by the gingival cells. In particular, it is said that about 60% of the constituents of periodontal tissue are collagen. When gingival cells are damaged by bacterial infection, inflammation, etc., gingival fibroblasts and immunocompetent cells excessively produce collagenase (collagenase) for an infection defense reaction, and as a result, collagen is decomposed. It causes gingival tissue destruction and gingival retraction (shrinkage, receding). In addition, it is known that bacteria themselves that have escaped infection defense reactions such as immune responses also produce collagen-degrading enzymes and trypsin-like enzymes, and as a result, enable tissue destruction and further invasion into tissues (non-patent documents). 1). Destruction and regression of gingival tissue by such collagen-degrading enzymes lead to the progression of periodontal disease and become a factor of tooth loss.

A group of metal enzymes that act on an extracellular matrix such as collagen to promote its degradation is generally called a matrix metalloprotease (also simply referred to as "MMP" in the present specification). Of these, collagen-degrading enzymes (collagenase: MMP-1 and MMP-8) that promote the decomposition of collagen have been reported to be expressed around the adherent epithelium of periodontal tissue and the periodontal pocket epithelium (non-patent). Document 2). From this, it can be seen that the increase in the expression and production of MMP and the increase in activity are closely related to the above-mentioned destruction of gingival tissue, gingival retraction, and progression of the pathological condition of periodontal disease. As a matter of fact, the amount of MMP contained in the oral lavage fluid, gingival crevicular fluid and saliva of periodontal disease patients reflects the pathological condition of periodontal disease patients. It has also been reported that the amount has decreased (Non-Patent Document 3). In addition, when the amount of MMP in gingitis patients and healthy subjects was measured, it was reported that the amount of MMP possessed by gingitis patients was very large, and that it was remarkable in periodontal disease patients (non-). Patent Document 4). From this, it is considered that a substance that suppresses the production of MMP in the periodontal tissue or a substance that inhibits the enzymatic activity of MMP is useful for suppressing the progression of the pathological condition of the periodontal tissue associated with local inflammation and its exacerbation. Conceivable.

By the way, hinokitiol has been conventionally known to have antibacterial action, anti-inflammatory action, tissue astringent action and the like, and has been added to oral compositions for the purpose of prevention and treatment of gingival inflammation and periodontal disease. Further, it is known that it has a hair follicle function activating action (Patent Document 1) and has an action of enhancing the skin cell proliferation action of psychosaponin (Patent Document 2). However, hinokithiol has the effect of suppressing the production of collagenase-degrading enzyme (fibroblast collagenase: MMP-1), which is a type of matrix metalloproteinase, and Porphyromonas is considered to be a promising causative agent of adult periodontal disease. It was not known that it has an effect of suppressing the increase in MMP-1 production caused by the infection of Lomonas gingivalis (P. gingivalis).

JP-A-2002-47145 Japanese Unexamined Patent Publication No. 5-262635

Takeshi Kikuchi et al., "Immunological Consideration on Tissue Destruction in Periodontal Disease," Jpn J Gastroenterol Surg, 59 (4): 185-190, 2017 M. Kylmaniemi et al., J. Dent Res., 75, pp.919-926 (1996) M. Makela et al., J. Dent Res., 73, pp.1397-1406 (1994) A. Haerian et al., J. Clin Periodontol., 22, pp.505-509 (1995)

An object of the present invention is to provide a new use based on the new action found for hinokitiol. Specifically, it is an object of the present invention to provide an agent for suppressing gingival tissue destruction, an agent for suppressing or improving gingival recession, and an agent for suppressing the production of MMP-1 in periodontium, based on the action of hinokitiol.

In daily research, the present inventors have found that the increase in the amount of MMP-1 produced due to infection with periodontal disease bacteria in the periodontal tissue is suppressed and reduced by treatment with hinokitiol. Was newly found. It is believed that these actions of hinokitiol can protect the gingival tissue from destruction by MMP-1 and prevent or ameliorate gingival retraction (shrinkage or retraction). It is also possible to prevent the exacerbation of periodontitis and periodontal disease by protecting the gingival tissue from destruction by MMP-1. The present invention has been completed based on such findings, and includes the following embodiments.

Item 1. A gingival tissue destruction inhibitor characterized by containing hinokitiol as an active ingredient.
Item 2. An agent for suppressing or improving gingival recession, which comprises hinokitiol as an active ingredient.
Item 3. An agent for suppressing the production of collagen-degrading enzyme MMP-1 in periodontal tissue, which comprises hinokitiol as an active ingredient.
Item 4. A group consisting of an action of suppressing gingival tissue destruction, an action of suppressing or improving gingival recession, and an action of suppressing the production of collagenase degrading enzyme MMP-1 in the periodontium by blending hinokithiol into the oral composition. A method of using hinokithiol, which comprises imparting at least one action selected from. The method of use is for producing at least one oral composition selected from the group consisting of an agent for suppressing gingival tissue destruction, an agent for suppressing or improving gingival recession, and an agent for suppressing the production of MMP-1 in periodontal tissue. , Can be rephrased as the use of hinokithiol.

By treating the oral cavity with the gingival tissue destruction inhibitor of the present invention, it is possible to suppress an increase in the production of MMP-1 in the periodontium, and as a result, it is possible to suppress the destruction of the gingival tissue. Further, by treating the oral cavity with the gingival recession suppressing or improving agent of the present invention, the destruction of the gingival tissue by MMP-1 can be suppressed, and as a result, the shrinkage or retreat of the gingiva can be suppressed or improved. Can be done. Furthermore, by treating the oral cavity with the MMP-1 production inhibitor of the present invention, it is possible to suppress an increase in the production of MMP-1 in the periodontal tissue caused by infection with periodontal disease bacteria, and as a result, the gingival tissue. Destruction can be suppressed. In addition, the shrinkage and retraction of the gingiva can be suppressed or improved. As described above, according to the oral compositions for various purposes of the present invention, inflammation of the gingival tissue (including gingival inflammation) caused by MMP-1 and pathological conditions of the periodontal tissue (including periodontal disease) associated with the exacerbation thereof. ) Is useful for suppressing and improving the progress.

Human gingival fibroblasts (HGF-1) in the presence of lipopolysaccharide (LPS-PG) derived from Porphyromonas gingivalis in the absence (comparative example) or presence of hinokithiol (Example 1: 1 ppm, Example). After culturing under the condition of 2: 5 ppm), the production amount (pg / mL) of type 1 human fibroblast collagenase (MMP-1) was measured after 24 hours and 48 hours, and the result was measured per viable cell number. It is a figure which shows the conversion result. The figure shows the average value of n = 6.

The gingival tissue destruction inhibitor, the gingival recession inhibitory or ameliorating agent, and the MMP-1 production inhibitor in the periodontium of the present invention are all characterized by containing hinokitiol as an active ingredient.

Hinokitiol is a compound represented by the following formula.

Hinokitiol (2-hydroxy-4-isopropylcyclohepta-2,4,6-trien-1-one, also known as "β-thujaplicin") is found in essential oils such as Taiwan Hinoki and Aomori Hiba. It is a natural product contained in Thujaplicin, and although it has low toxicity, it has a wide antibacterial spectrum and is a crystalline substance having an excellent antibacterial action. A salt is formed when it comes into contact with a metal ion, and in the present invention, the salt is also collectively referred to as hinokitiol. Hinokitiol can be isolated from the essential oils extracted from the above-mentioned Taiwan cypress and Hiba, but recently, it is also produced and provided by chemical synthesis. The hinokitiol used in the present invention may be of natural origin or synthetic product. Further, the essential oil or a crude product of the essential oil may be used regardless of the refined product as long as the effect of the present invention is not impaired.

The blending amount of hinokithiol in the gingival tissue destruction inhibitor, gingival recession inhibitor or ameliorating agent of the present invention, or the MMP-1 production inhibitor in periodontal tissue (hereinafter, these are collectively referred to as "this agent") is The amount of hinokithiol that exerts each effect when each agent is applied to the oral cavity as an oral composition is not particularly limited as long as it is sufficient. For example, when this drug is an oral composition to be applied to the oral cavity as it is, it can be appropriately selected and set from the range of 0.005 to 0.5% by mass. It is preferably 0.01 to 0.3% by mass, and more preferably 0.02 to 0.2% by mass. When this drug is an oral composition that is diluted with water at the time of use and applied to the oral cavity, it may contain hinokitiol so that the concentration after dilution is within the above range. For example, although not limited, when used, for example, diluted 10-fold with water at the time of use, the oral composition may be prepared so as to contain, for example, hinokitiol at a concentration of 0.05 to 5% by mass. it can. Furthermore, this drug has an effect of suppressing gingival tissue destruction, an effect of suppressing or improving gingival recession, or an effect of suppressing the production of MMP-1 in the periodontium in the above-mentioned oral compositions (including those used as they are and diluted before use). When used as an additive for imparting, the content of hinokithiol is not particularly limited as long as the oral composition can be prepared, for example, from the range of 0.1 to 100% by mass. It can be set as appropriate. Incidentally, the effect of suppressing gingival tissue destruction on gingival tissue destruction is due to the action of hinokithiol, which reduces and suppresses the increase in MMP-1 production in periodontal tissue due to periodontal disease infection. As shown in, the target human gingival fibroblasts were described in P.I. It can be evaluated by treatment with a gingival epithelial cell activator in the presence of gingivalis or its lipopolysaccharide (LPS). When the amount of MMP-1 expressed and produced in human gingival fibroblasts is reduced by treatment with gingival tissue destruction suppression as compared with the case without treatment with gingival tissue destruction suppression, gingival tissue destruction suppression As a result, the increase in MMP-1 production in the periodontal tissue due to periodontal disease infection is reduced and suppressed, and the suppression of gingival tissue destruction can suppress the destruction of gingival tissue by MMP-1, that is, the suppression of gingival tissue destruction. It can be judged that it is effective. Further, the gingival recession suppressing or improving effect of the gingival recession suppressing or improving agent can be determined by the same method. The improving effect is an effect obtained by suppressing the gingival recession and returning (recovering) the gingival tissue to a healthy state over time. Gingival retraction includes gingival shrinkage and / and retraction. Further, the effect of suppressing the production of MMP-1 in the periodontal tissue of the present invention can be determined by the same method.

The gingival tissue destruction inhibitor, the gingival recession inhibitory or ameliorating agent, or the MMP-1 production inhibitor in the periodontium of the present invention is an oral composition for suppressing gingival tissue destruction, an oral composition for suppressing or ameliorating gingival recession, respectively. Alternatively, by applying it in the oral cavity as an oral composition for suppressing the production of MMP-1 in the periodontal tissue, each effect can be exhibited. As described above, this agent may be used as it is or diluted with water as an oral composition to be applied into the oral cavity, or it may be added to these oral compositions to make gingival tissue. It may be used as an additive for preparing an oral composition having an action of suppressing destruction, an action of suppressing or improving gingival recession, or an action of suppressing the production of MMP-1 in the periodontium. That is, this drug may be finally used in the form of an oral composition, and can be prepared and provided in a form suitable for it.

Since hinokitiol is a crystal that is slightly soluble in water, for example, when this drug is used as an additive to an oral composition, hinokitiol is prepared in a solution state in which it is dissolved in a soluble solvent such as ethanol or propylene glycol. It is preferable to be done.

Examples of the form of the oral composition include any form such as tablets, pills, granules, liquids, suspensions, emulsions, sheets, gels, foams and pastes. These forms include, specifically, dentifrices (including powder, liquid, cream, and paste formulations), mouthwashes (including liquid and foam formulations), ointments, and pastilles. Agents, gingival cream, gingival gel, artificial tooth stabilizers, troches, chewing agents (including chewing gum, gummy, etc.), patches (films, packs), candies (candy, tablet confectionery [cool confectionery, mint tablets, supplements] Including] etc.) etc. are included. By preparing such a form and applying it in the oral cavity, it is possible to effectively suppress the increase in MMP-1 production, and as a result, the destruction of the gingival tissue by MMP-1 is suppressed, and the involution of the gingiva can be suppressed or improved. .. These actions prevent the inflammation of the gingiva and the exacerbation of periodontal disease, and by making it healthy, the oral cavity related to periodontal tissues such as periodontitis, gingival inflammation, periodontitis, pericoronitis, and pericoronitis. It becomes possible to effectively deal with the prevention or treatment of internal diseases.

These oral compositions can contain various components used in the art according to each form, and do not limit them as long as they do not interfere with the effects of the present invention. For example, in the case of dentifrices, abrasives, wetting agents, binders, foaming agents, thickeners, surfactants, sweeteners, fragrances, coloring agents, preservatives, etc., as long as they do not interfere with the effects of the present invention. Medicinal ingredients (eg, anti-inflammatory agents, etc.) can be blended.

[Abrasive agent] Silica-based abrasives such as precipitated silica, silica gel, aluminosilicate, and zirconosilicate, calcium hydrogen phosphate for toothpaste, calcium dibasic dihydrate, dicalcium phosphate anhydride, phosphorus such as calcium pyrophosphate, etc. Acid-based abrasives, aluminum hydroxide, alumina, titanium dioxide, crystalline zirconium silicate, polymethylmethacrylate, insoluble calcium metaphosphate, light calcium carbonate, heavy calcium carbonate, magnesium carbonate, magnesium tertiary phosphate, zeolite, silicic acid Zirconium, hydroxyapatite, fluoroapatite, calcium-deficient apatite, calcium phosphate tribasic, calcium tetraphosphate, calcium eighth phosphate, synthetic resin-based abrasive.

[Wetting agent] Polyhydric alcohols such as glycerin, concentrated glycerin, diglycerin, sorbitol, maltitol, dipropylene glycol, propylene glycol, 1,3-butylene glycol, xylitol and the like.

[Binder] Cellulose-based binders such as sodium carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxymethyl ethyl cellulose, and methyl cellulose, xanthan gum, carrageenan, guagam, sodium alginate, cationized cellulose, montmorillonite, gelatin, Sodium polyacrylate, etc.

[Effervescent agent] N-acyl amino acid salts such as sodium lauryl sulfate, sodium lauroyl sarcosin, sodium alkyl sulfosuccinate, sodium coconut oil fatty acid monoglycerin sulfonate, sodium α-olefin sulfonate, N-acylglutamate, 2-alkyl-N -Carboxymethyl-N-Hydroxyethyl imidazolinium betaine, multitoll fatty acid ester, sucrose fatty acid ester, polyglycerin fatty acid ester, fatty acid diethanolamide, polyoxyethylene sorbitan monostearate, polyoxyethylene hydrogenated castor oil, polyoxyethylene Fatty acid ester, alkyl glycoside, etc.

[Consolidating agent] Glycerin, sorbitol, propylene glycol, polyethylene glycol having a molecular weight of 200 to 6000, ethylene glycol, 1,3-butylene glycol, reduced starch saccharified product, etc.

[Surfactant] Anionic surfactant, nonionic surfactant, amphoteric surfactant, cationic surfactant and the like can be blended. Examples of the anionic surfactant include sodium alkyl sulfate such as sodium lauryl sulfate and sodium myristyl sulfate, sodium N-acylsarcosine such as sodium N-lauroyl sarcosin and sodium N-myristyl sarcosin, sodium dodecylbenzene sulfonate, and hydrogenated coconut fatty acid. Monoglyceride N-acylglutamate such as sodium monosulfate, sodium lauryl sulfoacetate, sodium N-palmitoyl glutamate, sodium N-methyl-N-acyltaurine, sodium N-methyl-N-acylalanine, sodium α-olefin sulfonate, etc. Can be mentioned. Examples of nonionic surfactants include polyoxyethylene hydrogenated castor oil, sucrose fatty acid ester, alkylol amide, polyoxyethylene sorbitan monostearate, polyoxyethylene polyoxypropylene glycol, alkyl glucoside, and decaglyceryl laurate. Used. Examples of amphoteric surfactants include lauryldimethylaminoacetate betaine, N-palm oil fatty acid acyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine, coconut oil fatty acid amide propyl dimethylaminoacetate betaine, and coconut oil fatty acid amide propyl. Betaine etc.

[Sweetness] Saccharin sodium, aspartame, stebioside, stevia extract, paramethoxycinnamic aldehyde, neohesperidyl dihydrochalcone, perillartine, sucralose, xylitol, reduced palatinose, erythritol, maltitol, etc.

[Fragrance] Peppermint oil, sparemint oil, anis oil, eucalyptus oil, winter green oil, cassia oil, clove oil, thyme oil, sage oil, lemon oil, orange oil, peppermint oil, cardamon oil, coriander oil, mandarin oil, lime Oil, lavender oil, rosemary oil, laurel oil, camomill oil, caraway oil, majorum oil, bay oil, lemongrass oil, origanum oil, pine needle oil, neroli oil, rose oil, jasmine oil, iris concrete, absolute peppermint , Absolute rose, orange flower and other natural fragrances, and processed fragrances (pre-reservoir cut, rear-reservoir cut, distilling, liquid extraction, essence, powder fragrance, etc.) and menthol. , Carvone, Anetol, Cineol, Methyl salicylate, Synamic aldehyde, Eugenol, 3-l-mentoxypropane-1,2-diol, Timor, Linarol, Linaryl acetate, Limonen, Menton, Menthyl acetate, N-substituted-Peppermint -3-Carboxamide, Peppermint, Octylaldehyde, Citral, Pregon, Calvia Acetate, Anisaldehyde, Ethylacetate, Ethylbutyrate, Allylcyclohexanepropionate, Methylanthranilate, Ethylmethylphenylglycidate, Vanillin, Undecalactone , Hexanal, propyl alcohol, butanol, isoamyl alcohol, hexenol, dimethylsulfide, cycloten, furfural, trimethylpyrazine, ethyllactate, methyllactate, ethylthioacetate and other single flavors, as well as strawberry flavor, apple flavor, banana flavor, pineapple flavor. , Grape flavor, Mango flavor, Butter flavor, Milk flavor, Fruit mix flavor, Tropical fruit flavor, etc.

[Colorant] Blue No. 1, Yellow No. 4, Green No. 3, etc.

[Preservatives] Parabens such as methylparaben, ethylparaben, propylparaben and butylparaben, sodium benzoate, phenoxyethanol, alkyldiaminoethylglycine hydrochloride and the like.

[Medicinal Ingredients] Fluorine compounds such as sodium fluoride, sodium monofluorophosphate, tin fluoride, water-soluble phosphate compounds such as potassium salt of orthophosphate and sodium salt, sodium polyphosphate, tranexamic acid, epsilon-aminocaproic acid , Allantin, Allantin Chlorhydroxyaluminum, Ascorbic acid, dl-α-tocopherol acetate, dl-α-tocopherol nicotinate, pyridoxin hydrochloride, dihydrocholesterol, α-bisabolol, chlorhexidine salts, sodium azulene sulfonate, guaizrene sulfonic acid, Copper compounds such as glycyrrhetin, glycyrrhetinic acid, copper gluconate, aluminum lactate, strontium chloride, potassium nitrate, velverine, hydroxamic acid and derivatives thereof, zeolite, maleic anhydride copolymer, polyvinylpyrrolidone, polyethylene glycol, epidihydrocholesterin , Cetylpyridium chloride, benzethonium chloride, isopropylmethylphenol, sodium lauroyl sarcosin, dihydrocholesterol, trichlorocarbanilide, zinc citrate, soft touki extract, sardine extract, chamomile, chowji, rosemary, ginger, benibana , Resoteam chloride, sodium chloride, etc.

The blending amount of these optional components can be a normal amount used in the art as long as the effects of the present invention are not impaired.

This drug, which is thus prepared in the form of an oral composition, can be treated in the oral cavity of a target subject by normal use according to the form. For example, when it has the form of a dentifrice, each effect of this drug can be exhibited by brushing the subject's teeth with this drug. In addition, when it has the form of a mouthwash, each effect of this drug can be exhibited by washing (rinsing) the oral cavity of the subject with this drug. The same applies when this drug has the other forms described above. Although not limited, in order to exert the effect of the present invention more, when applied into the oral cavity, the concentration of hinokitiol in the oral cavity including saliva is in the range of 1 nM to 20 mM, preferably 10 nM to 10 mM. It is preferably prepared to be 10 nM to 5 mM.

The number of times this drug is applied to the oral cavity is not particularly limited, and it can be applied once or multiple times a day, preferably after meals and / and before and after bedtime.

As described above, in the present specification, the terms "including" and "containing" include the meanings of "consisting of" and "consisting of substantially".

Hereinafter, the present invention will be described with reference to experimental examples in order to help understanding the structure and effects of the present invention. However, the present invention is not limited by these experimental examples. The following experiments were performed under room temperature (about 25 ° C.) and atmospheric pressure conditions, unless otherwise noted.

Experimental Example 1 Effect of hinokithiol on the proliferation of gingival epithelial cells Human gingival fibroblasts (HGF-1) were subjected to the presence or absence of hinokithiol in the presence of lipopolysaccharide (LPS-PG) derived from Porphyromonas gingivalis. After culturing under the conditions, the amount of type 1 human fibroblast collagenase (MMP-1) produced was measured over time, and the effect of hinokithiol on the expression and production of collagen-degrading enzyme in gingival cells was evaluated.

1. 1. Material cells: Normal human gingival fibroblasts (HGF-1 cells) (ATCC CRL-2014)
Medium: D-MEM High Glucose (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.)
Reagents for measuring live cells:
(1) MMP-1 ELISA kit (Human Total MMP-1 DuoSet ELISA, manufactured by R & D Systems)
(2) Cel Counting Kit-8 Cel (manufactured by Dojin Chemical Laboratory Co., Ltd.)
LPS-PG: Lipopolysaccharide derived from Porphyromonas gingivalis [Pg] (manufactured by Nacalai Tesque, Inc.)
Hinokitiol: Tokyo Chemical Industry Co., Ltd. Solvent for hinokitiol: Dimethyl sulfoxide 50 ppm, POE cured castor oil 50 ppm

2. Experimental method (1) Measurement of MMP-1 amount Normal human gingival fibroblasts (HGF-1 cells) were seeded on a 96-well plate containing a medium at ^{a ratio of 1 × 10 4} cells / well, and incubator (37). The cells were cultured for 24 hours (initial culture) in 5% CO _2). After the initial culture (24 hours), the medium was removed, the test sample solutions (Comparative Examples, Examples 1 and 2) shown in Table 1 and LPS-PG (final concentration 10 μg / mL) were added, and the incubator was further added. Incubate at (37 ° C., 5% CO ₂ ) for 24 hours or 48 hours (exposure culture). After 24 hours and 48 hours from the culture, the sample solution (culture solution) was collected, and the amount of MMP-1 (pg / mL) in the culture solution was measured by the ELISA method according to the manual of the MMP-1 ELISA kit.

(2) Measurement of the number of viable cells The number of viable cells in the sample solution (culture solution) collected after 24 hours and 48 hours of the culture was measured using the CCK kit. Specifically, first, for the sample (initial sample) obtained by the initial culture, the medium is removed from the well, the gingival fibroblasts are washed with PBS, and the CCK diluted solution (CCK solution: medium =) attached to the CCK kit is used. After adding 100 μL of 1:10) and allowing to _{stand in an incubator (37 ° C., 5% CO 2} ) for 1 hour, the absorbance at wavelengths of 450 nm and 650 nm was measured. For the sample (exposed sample) obtained by the exposure culture, the medium and the test sample solution were removed from the sample solution collected from the well, the gingival fibroblasts were washed with PBS, and the CCK diluted solution (same as above). Was added in 100 μL and allowed to stand in an incubator (37 ° C., 5% CO ₂ ) for 1 hour, and then the absorbance at wavelengths of 450 nm and 650 nm was measured. For the number of living cells in the exposed sample (sample solution), compare the _{measured difference between the measured absorbance at a wavelength of 450 nm and the absorbance at 650 nm (absorbance 450 nm} -absorbance _{650 nm} ) with the difference measured for the initial sample (absorbance _{450 nm} -absorbance _{650 nm).} And calculated.

3. 3. Results For Comparative Examples and Examples 1 and 2, the amount of MMP-1 (pg / mL) in the sample solution (culture solution) is shown in Table 2 and FIG. The values shown in Table 2 and FIG. 1 are all MMP- per viable cell number (relative value), where 1 is the number of viable cells contained in the sample after culturing the sample of the comparative example for 24 hours or 48 hours. It is converted to 1 amount (pg / mL). The result is the average value of the number of tests (n = 6).

As shown in Table 2 and FIG. 1, when lipopolysaccharide (LPS-PG) derived from Porphyromonas gingivalis was added to human gingival fibroblasts and cultured, in the absence of hinokithiol, in human gingival fibroblasts. It was confirmed that the amount of expression and production of the collagen-degrading enzyme MMP-1 in the above was increased with time (comparative example). On the other hand, by culturing human gingival fibroblasts in the presence of hinokitiol (Examples 1 and 2), the amount of MMP-1 cultured in the absence of hinokitiol (Comparative Example) was compared with MMP-1. It was confirmed that the production amount was significantly reduced. From this, it was found that hinokitiol has an effect of suppressing and reducing the amount of MMP-1 whose production is increased by bacterial infection in the gingival tissue. From this, according to hinokitiol, it is considered that by suppressing the overproduction of collagen-degrading enzyme due to bacterial infection, it is possible to prevent or improve the destruction of periodontal tissue and gingival recession caused by collagen-degrading enzyme.

[Prescription example]
According to the formulation below, the formulations of the present invention in the form of dentifrice (Table 3), mouthwash (Table 4), and gel (Table 5) can be prepared.

Claims (4)

A gingival tissue destruction inhibitor characterized by containing hinokitiol as an active ingredient. An agent for suppressing or improving gingival recession, which comprises hinokitiol as an active ingredient. An agent for suppressing the production of collagen-degrading enzyme MMP-1 in periodontal tissue, which comprises hinokitiol as an active ingredient. By blending hinokitiol into the oral composition, the oral composition is selected from the group consisting of an action of suppressing gingival tissue destruction, an action of suppressing or improving gingival recession, and an action of suppressing the expression or production of collagen degrading enzyme MMP-1. A method of using hinokitiol, which comprises imparting at least one action to be performed.