JP2004051603A - Agent for oral cavity - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new glucosyltransferase inhibitor having an excellent performance, a plaque formation inhibitor, an inhibitor for gingivitis and/or periodontal disease by using a natural material having a high safety and an oral cavity agent, a dental caries-preventing agent, a food or beverage, etc., imparted with the dental caries-preventing activity by adding them. <P>SOLUTION: The glucosyltransferase inhibitor, plaque formation inhibitor and gingivitis and/or periodontal disease inhibitor are characterized by containing a β-glucan and/or mevalonic acid as active ingredients. And further, by adding them, the oral cavity agent, caries-preventing agent, and food and beverage imparted with the caries-preventing activity are provided. <P>COPYRIGHT: (C)2004,JPO

Description

【００９７】
（２）唾液緩衝能の測定
唾液緩衝能を測定可能なＤｅｎｔｏｂｕｆｆ　Ｓｔｒｉｐ　（株式会社モリムラ製）を用いて行った。Ｄｅｎｔｏｂｕｆｆ　Ｓｔｒｉｐ　によれば、唾液緩衝能を四段階に評価することができる。具体的な測定は、まず、唾液サンプルをピペットで取り、ストリップ上に落とし、５分後、ストリップの色とカラーチャートを比較した。なお、このキットにおいて、唾液の緩衝能は大きい順に、即青、青、緑、そして黄色のレベルに分類される。結果を表２に示す。
【００９８】
【表２】
表２

【００９９】
（３）ｍｕｔａｎｓ　ｓｔｒｅｐｔｏｃｏｃｃｉ　の菌数の測定
口腔内のｍｕｔａｎｓ　ｓｔｒｅｐｔｏｃｏｃｃｉの菌数をＤｅｎｔｏｃｕｌｔ−ＳＭキット（株式会社モリムラ製）を用いて行った。ここでＤｅｎｔｏｃｕｌｔ−ＳＭは、唾液中のｍｕｔａｎｓ　ｓｔｒｅｐｔｏｃｏｃｃｉの菌数を調べることができる試験培地である。まず、キット中のバシトラシン錠を加えた培養液に、舌上で回転させたストリップを入れ、３７℃で、２日間培養した。ｍｕｔａｎｓ　ｓｔｒｅｐｔｏｃｏｃｃｉは、ストリップ上に、青色のコロニーとして表れ、ストリップ上のコロニーの密度と、モデルチャートを比較して、唾液１ｍｌ中の菌数を、それぞれおよそ、０個を◎、１０万個のレベルを○、５０万個を△、１００万個を×として判定した。
【０１００】
【表３】
表３

【０１０１】
（４）Ｌａｃｔｏｂａｃｉｌｌｕｓ　の菌数の測定
口腔内のＬａｃｔｏｂａｃｉｌｌｕｓの菌数の測定をＤｅｎｔｏｃｕｌｔ−ＬＢキット（株式会社モリムラ製）を用いて行った。ここで、Ｄｅｎｔｏｃｕｌｔ−ＬＢは、唾液中のＬａｃｔｏｂａｃｉｌｌｕｓの菌数を調べることができる試験培地である。まず、唾液サンプルをキット中に培地が含浸されたスライドの両面に流し、３７℃で、４日間培養した。Ｌａｃｔｏｂａｃｉｌｌｕｓは、スライド上に、白色のコロニーとして表れ、スライド上のコロニーの密度と、モデルチャートを比較して、その菌数を判定した。
【０１０２】
結果を表４に示す。表中、唾液１ｍｌ中の菌数がそれぞれおよそ１０００個（◎で示す）、１万個（○で示す）、１０万個（△で示す）、および１００万個（×で示す）の各レベルを意味する。
【０１０３】
【表４】
表４

【０１０４】
〔試験２〕：口腔内細菌Ｃａｎｄｉｄａの測定
口腔内のＣａｎｄｉｄａの菌数を、Ｏｒｉｃｕｌｔ−Ｎ（株式会社モリムラ製）を用いて調べた。ニッカーソン培地を塗布したスライドの両面に採取した唾液サンプルを流し、３７℃で、２日間培養した。Ｃａｎｄｉｄａは、スライド上に、褐色のコロニーとして表れ、スライド上のコロニーの密度と、モデルチャートを比較して、その菌数を判定した。
【０１０５】
結果を表５に示す。表中、唾液１ｍｌ中の菌数がそれぞれおよそ１０００個（◎で示す）、１万個（○で示す）、１０万個（△で示す）、および１００万個（×で示す）の各レベルを意味する。
【０１０６】
【表５】
表５

【０１０７】
〔試験３〕：歯周病関連要因の評価
成人性歯周炎の主たる病原菌といわれるＰｏｒｐｈｙｒｏｍｏｎａｓ　ｇｉｎｇｉｖａｌｉｓ、Ｔｒｅｐｏｎｅｍａ　ｄｅｎｔｉｃｏｌａ、およびＢａｃｔｅｒｏｉｄｅｓ　ｆｏｒｓｙｔｈｕｓを、それが産生する酵素を特異的に検出する歯周病原菌検査薬（ペリオチェック、サンスター株式会社製）によって調べた。まず、歯周ポケットの最深部に３０秒間挿入したペーパーポイントを、基質・色源体剤を含むバイアル内に直接投入し、酵素溶液を加えた。３７℃で、１５分間、インキュベートした後、青色変化の程度を、標準色調（色見本）と比較して、これらの歯周病原菌由来のペプチターゼ活性を測定した。
【０１０８】
結果を表６に示す。表中、（−）は陰性、（＋）は陽性、（＋＋）は強陽性を意味する。
【０１０９】
【表６】
表６

【０１１０】
【発明の効果】
本発明によれば、優れた性能を有するグルコシルトランスフェラーゼ阻害剤、プラーク形成抑制剤、歯肉炎及び／または歯周病抑制剤、さらにはそれらを添加して、う蝕予防作用を付与した口腔用剤、う蝕予防剤、飲食物等を提供することができる。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a glucosyltransferase inhibitor having an action of inhibiting the activity of a glucosyltransferase, an enzyme for synthesizing glucan that causes caries, to prevent the occurrence of tooth decay, and to inhibit the formation of plaque, which is a place where oral microorganisms grow, The present invention relates to a plaque formation inhibitor, a gingival salt, a periodontal disease inhibitor, an oral agent and a food and drink provided with a caries preventive action, which have an action of preventing periodontal disease. Further, in this specification, foods and drinks are meant to include not only foods and drinks for humans but also feeds or feeds for livestock and companion animals that may cause dental caries.
[0002]
[Prior art]
In the development of dental caries, microorganisms in the oral cavity, particularly enzymes, glucosyltransferases produced by Streptococcus mutans, are involved. That is, a part of the sucrose in the food and drink remaining in the oral cavity is converted into a water-insoluble and strongly adherent glucan by the action of glucosyltransferase, which adheres to the surface of the tooth together with the oral microorganisms to form plaque (tooth). To form dirt). Microbes in the dental plaque metabolize sugars in the food to form acids, and this acid demineralizes and erodes tooth enamel.
[0003]
Therefore, in order to prevent dental caries, plaque attached to the tooth surface is not only removed using toothpaste, but also glucan production is inhibited by inhibiting the growth of Streptococcus mutans and the action of glucosyltransferase in the oral cavity. It is most effective to prevent them and thus prevent plaque from forming.
[0004]
From such a viewpoint, in recent years, oral preparations and foods and drinks provided with a caries-preventing action by including a substance having a glucosyltransferase inhibitory action and a substance suppressing plaque formation have been provided. Glucosyltransferase inhibitors conventionally known as suitable for such uses include mutastain, crude drug tannins, ellagic acid, green tea polyphenol, oolong tea extract and the like. In addition, on oral hygiene, to prevent or treat periodontal disease such as gingivitis or alveolar pyorrhea caused by dirty conditions such as food residues and propagation of various microorganisms, or to promote the secretion of sleep fluid, Ethanol, various germicides, anti-inflammatory agents and the like have been used, but their performance has not been sufficient.
[0005]
[Problems to be solved by the invention]
An object of the present invention is to provide a novel glucosyltransferase inhibitor, a plaque formation inhibitor, a gingivitis and / or periodontal disease inhibitor, which has excellent performance, using a highly safe natural product as a raw material, and further comprises adding them. In addition, it is an object of the present invention to provide an oral agent, a caries preventive agent, a food and drink, etc. having a caries preventive action.
[0006]
[Means for Solving the Problems]
As a result of intensive studies, the present inventors have found that β-glucan and mevalonic acid have a caries-preventing action and a periodontal disease-preventing action, and have completed the present invention.
[0007]
That is, the present invention provides a glucosyltransferase inhibitor comprising β-glucan and / or mevalonic acid as an active ingredient.
[0008]
The present invention also provides a plaque formation inhibitor comprising β-glucan and / or mevalonic acid as an active ingredient.
[0009]
The present invention also provides a gingivitis inhibitor and / or a periodontal disease inhibitor, which comprises β-glucan and / or mevalonic acid as an active ingredient.
[0010]
The present invention also provides the glucosyltransferase inhibitor, the plaque formation inhibitor, the gingival salt inhibitor and / or the periodontal disease inhibitor wherein the β-glucan is derived from cereals, microorganisms and / or basidiomycetes. It is.
[0011]
The present invention also provides an oral agent characterized by containing one or more selected from the group consisting of the glucosyltransferase inhibitor, plaque formation inhibitor, gingivitis inhibitor, and periodontal disease inhibitor. Things.
[0012]
Further, the present invention is characterized by containing one or more selected from the group of the glucosyltransferase inhibitor, plaque formation inhibitor, gingivitis inhibitor, periodontal disease inhibitor, a caries preventive agent or It is intended to provide caries-preventing food and drink.
[0013]
BEST MODE FOR CARRYING OUT THE INVENTION
In the present invention, β-glucan is a kind of polysaccharide, and includes a 1-2-β-D-glucopyranose bond, a 1-3-β-D-glucopyranose bond, and a 1-4-β-D-glucopyranose bond. , Which has at least two or more types of 1-6-β-D-glucopyranose bonds, and is selected from the group consisting of β-glucan derived from grains, β-glucan derived from microorganisms, and β-glucan derived from basidiomycetes. Or two or more types are mentioned as preferable examples.
[0014]
Explaining β-glucan derived from cereals, the cereals are preferably grasses. Examples of grasses include grains such as rice, wheat, corn, sorghum, barnyardgrass, millet, millet, barley, oats (crow oats), and rye. it can. In particular, β-glucan obtained by extraction from these grasses is preferable. In the present invention, β-glucan obtained by extraction is also referred to as extracted β-glucan.
[0015]
The β-glucan extracted from the gramineous plant is not particularly limited in its extraction method, and may be extracted by adding an extraction solvent to a gramineous plant as an extraction raw material. In addition, the extract itself in the case of solid-liquid separation, or a liquid or solid state obtained by concentrating β-glucan extracted from the extract by a known method, or by purifying the extract from the extract by a known method to purify the purity. It is possible to use those obtained by any production method such as the above-mentioned liquid or solid, any form, and any purity. Of course, there is no problem even if the extracted components other than β-glucan are mixed. In the present invention, these are all referred to as β-glucans extracted from grasses.
[0016]
For the extraction, the whole plant can be used as a raw material, but it is preferable to use a seed having a relatively high β-glucan content. Any of bran, bran, malt, germ, and endosperm parts obtained in the cereal purification step may be used, including whole pulverized matter (whole grain). Preferably the barley or oats whole grain or oats crushed from the outer periphery of the endosperm portion and the resulting bran, rice bran, bran or germ of wheat or corn, more preferably barley or oats It is the endosperm portion obtained by milling whole grains and grains from the outer periphery and bran generated at that time.
[0017]
Further, β-glucan extracted from a gramineous plant has a 1-2-β-D-glucopyranose bond, a 1-3-β-D-glucopyranose bond, a 1-4-β-D-glucopyranose bond, Β-glucan having at least two kinds of -6-β-D-glucopyranose bonds is preferable, and β-glucan consisting of 1-3,1-4-β-D-glucopyranose bond is preferably contained.
[0018]
Explaining the method of extracting β-glucan from a gramineous plant, β-glucan in a gramineous plant can be dissolved as an aqueous solution as a water-soluble polymer.For example, water, hot water, hot water Alternatively, using a salt solution, furthermore, an acid, an alkaline aqueous solution, an organic solvent, or the like, extraction can be performed at an arbitrary temperature at an arbitrary temperature for 2 to 100 times the amount of powder with respect to powder. Further, β-glucan can be obtained by solid-liquid separation of the extract. Among these, β-glucan extracted with water, hot water or hot water is preferable, and β-glucan extracted with hot water having a temperature of 80 ° C. or lower and 4 ° C. or higher is more preferable. Further, an extraction accelerator may be added at the time of extraction.
[0019]
Specifically, as a method for obtaining a high molecular weight β-glucan from barley, for example, a method of producing a waxy barley as a raw material by water extraction (Japanese Patent Publication No. 4-11197), or a method of producing barley, oats Of β-glucan having a weight-average molecular weight of 100,000 to 1,000,000 by alkali extraction, neutralization, and alcohol precipitation (JP-B-6-83652), using barley bran having a milling yield of 82% or less as a raw material Β-glucan obtained by a method of extracting β-glucan with hot water at 80 to 90 ° C. (Japanese Patent Application Laid-Open No. H11-225706), and β-glucan obtained by these production methods are further purified by known methods. What was made into low molecular weight β glucan. For example, any known polysaccharide hydrolysis reaction can be used as a method for reducing the molecular weight. For example, it is known that a water-soluble polysaccharide is hydrolyzed by heating under pressure in the presence of an acid, and this can be used to reduce the molecular weight. It is also effective to reduce the molecular weight using a hydrolysis reaction by an enzyme. As the enzyme, 1,3-β-glucanase or the like can be used. Furthermore, β-glucan obtained by directly extracting from raw cereal grains by a method described in WO98 / 13056, JP-A-2002-97203 or the like can also be used. Moreover, you may use the extraction promoter etc. of Unexamined-Japanese-Patent No. 2002-105103.
[0020]
The β-glucan derived from cereals used in the present invention is a polymer, and β-glucan having any weight-average molecular weight can be used.However, since water solubility increases as the molecular weight decreases, the molecular weight is 3,000,000 or less, preferably It is preferably 500,000 or less, more preferably 100,000 or less. For example, β-glucan obtained by extraction from a grass plant may be reduced in molecular weight by a known method so as to improve water solubility, or β-glucan having a low molecular weight may be directly extracted.
[0021]
Explaining β-glucan derived from microorganisms, microorganisms are cells that themselves contain a large amount of β-glucan in their cell walls, and are cultured by inoculating microorganisms into their respective growth media and growing cells. The cells can be used as they are. Also, a cultured cell wall residue obtained by crushing cultured cells and removing the contents, or β-glucan extracted from a cultured cell or a cultured cell wall residue obtained by crushing the cultured cells and removing the contents Can be used as it is or purified, but it is preferable to use the extracted β-glucan.
[0022]
It is also possible to use β-glucan secreted and produced outside the cells, and to use the β-glucan after the completion of the culture as it is or after isolating and purifying the β-glucan from the culture. Although it can be used, it is preferable to use β-glucan after performing an operation of isolating and purifying β-glucan from the culture solution.
[0023]
Β-glucan derived from microorganisms is preferably used as it is or purified from β-glucan extracted from cultured cell or cultured cell wall residue obtained by crushing the cultured cell and removing the contents. It is most preferable to use β-glucan secreted and produced outside the cells together with the culture solution or purified from the culture solution.
[0024]
As microorganisms suitable for obtaining β-glucan derived from microorganisms, microorganisms conventionally used for food have high safety and are suitable. That is, yeasts, lactic acid bacteria, natto bacteria, acetic acid bacteria, koji molds, algae such as chlorella and spirulina, and microorganisms belonging to the genus Aureobasidium.
[0025]
Examples of yeast include yeast classified in the genus Saccharomyces used in alcohol brewing and baking processes such as beer, shochu, sake, wine, whiskey, etc., yeasts used in pressed bread yeast, and yeasts used in soy sauce brewing. And Candida genus used for microbial protein production.
[0026]
Examples of lactic acid bacteria include bacterium Lactobacillus and Bifidobacterium, staphylococci Leuconostoc, Pediococcus, Streptococcus, and Streptococcus. ) Genus is commonly used, and other lactic acid bacteria utilizing the genus Enterococcus, the genus Vagococcus, the genus Carnobacterium, the genus Aerococcus, or the genus Tetragenococcus. be able to. Specific lactic acid bacteria strains include Lactobacillus bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus lactis, Lactobacillus casei (L. casei), Lactobacillus brevis (L. brevis), Lactobacillus plantarum (L. plantarum), Lactobacillus salmon (L. sake), Streptococcus thermophilus, Streptococcus, Lactobacillus (S. lactis), Streptococcus cremoris (S. cremoris), Bifidobacterium longum (B Fidobacterium longum, Bifidobacterium bifidum (B. bifidum), Bifidobacterium breve (B. breve), Bifidobacterium infantis (B. infantis), Leuconostoc cremoris, Leuconostoc cremoris Stock Mecenteroides (Ln. Meseneroides), Leuconostoc oenos (Ln. Oenos), Pediococcus acidilactici, Pediococcus cerevisiae (P. cerevisiae), Pediococcus sp. And one or more types of conventionally used lactic acid bacteria such as C. pentosaceus) can be used. These may be used alone or in combination of two or more. Alternatively, the culture of Bifidobacterium and the culture of other lactic acid bacteria may be separately cultured and mixed.
[0027]
As the microorganism belonging to the genus Aureobasidium, any microorganism may be used as long as it is a strain that produces a glucose polymer having a β bond outside the cell by culturing the microorganism, and examples thereof include aureobasidium. A strain of Aureobasidium pullulans, specifically, IFO4466, IFO6353, IFO7757, ATCC9348, ATCC3092, ATCC4333023, or the like can be used.
[0028]
In addition, Bacillus strains of Bacillus natto, strains of acetobacterium Acetobacter, acetobacilli, Aspergillus aspergillus, Aspergillus, Penicillium, algae such as chlorella and spirulina, dried chlorella Xanthomonas which is known to produce a thickening polysaccharide used as a strain of the genus Aureobasidium, which is known to secrete and produce powder and pullulan extracellularly, and other food additives Xanthomonas genus, Aeromonas genus, Azotobacter genus, Alcaligenes genus, Erwinia genus, Enterobacter obactor) genus, sclerotinia Umm (Sclerotium) genus Pseudomonas (Pseudomonas) genus, Agrobacterium (Agrobacterium) genus, mention may be made of Makurohomopushisu (Macrophomopsis) strain of the genus.
[0029]
To explain β-glucan derived from basidiomycetes, basidiomycetes contain a large amount of β-glucan in the sclerotium where fruit bodies and hyphae aggregate in a lump, and are finely pulverized or extracted from pulverized material. And β-glucan purified from an extract can be used as basidiomycete-derived β-glucan. In addition, cultured cells obtained by germinating spores of basidiomycetes, inoculating mycelium into the respective growth media, and growing the cells can be used as they are. Alternatively, it is extracted from the cultured cell wall residue obtained by crushing the cultured cells and removing the contents, or the cell wall residue obtained by crushing the mycelium cultured cells or the mycelium cultured cells and removing the contents. The β-glucan derived from basidiomycetes can be used as it is or purified. When β-glucan secreted and produced outside the cells is used, the culture solution can be used as it is, or β-glucan separated and purified from the culture medium can be used as the β-glucan derived from basidiomycete.
[0030]
Of these, the fruiting bodies and sclerotia were pulverized, and the β-glucan extracted from them was obtained as it was or purified, by crushing the mycelium cultured cells or mycelium cultured cells and removing the contents. It is preferable to use β-glucan extracted from the cell wall residue as it is, or to use it after purification.Furthermore, it is preferable to use β-glucan secreted and produced outside the cells together with the culture solution or purified from the culture solution. Even more preferred.
[0031]
Cultivars are most preferred as basidiomycetes, but β-glucans from terminal fungi that are not subjected to commercial production can also be used in the present invention. Examples include Agaricus blazei, Amigasatatake, Amitake, Ezokaritake mushroom, Enokitake, Kanzotake mushroom, Kikurage, Kinugasataketake, Kuritake, Saketakebaketake, Sasakurehiyoketake, Coral Haritake, Shiitake, Shoro, Shirotaketake mushroom, Shirotake mushroom, Shirotake mushroom Midsummer in winter, nameko, naratake, naratakemodoki, shimeji mushroom, nikawaurokotake, nikawaharitake, numerisugitake, numerisugitamomodoki, hatstake, hiratake, bukurohyo, kurotake, bunashimeji, bunashimaritake, matsutake mushroom, matsutake mushroom , Mukitake, purple mushroom, yamadoritake, yamabushitake, willow matsutake, and the like.
[0032]
The method of isolating the cell wall residue of microorganisms and basidiomycetes as β-glucan is to add an appropriate amount of solvent to cultured microbial fungi, cultured mycelium, or cultivated sclerotia or fruiting bodies, and to autolyze or hydrolyze The cell wall residue can be isolated as β-glucan by destroying a part of the cell wall and removing the contents by the addition of, and collecting the residual component. In addition, damage to cells of microbial fungi and basidiomycetes by the physical force of a French press or ultrasonic crusher, etc., destroy some of them, remove the contents, and collect the residue to obtain β glucan Can be.
[0033]
There is no particular limitation on the method for extracting β-glucan of microorganisms and basidiomycetes, and the extraction may be performed by adding an extraction solvent to the microorganisms and basidiomycetes used as extraction raw materials. As the extraction solvent, one or a mixture of two or more of water, a salt solution, an aqueous acid solution, an aqueous alkali solution, an organic solvent, and the like can be used. The extraction efficiency can be increased by using an enzyme that degrades the cell wall in combination. The extract may be an extract itself after solid-liquid separation, or a liquid or solid form obtained by concentrating β-glucan extracted from the extract by a known method, or purified by a known method from the extract. It is possible to use those obtained by any production method, such as liquid or solid substances with increased purity, any form, and any purity. Of course, there is no problem even if the extracted components other than β-glucan are mixed. In the present invention, these are all referred to as β-glucans extracted from microorganisms and basidiomycetes. Further, a method of extracting β-glucan from microorganisms and basidiomycetes will be described.The β-glucan according to the present invention can be dissolved as an aqueous solution as a water-soluble polymer, and is, for example, a basidiomycete and is generally commercially available. Dried mushrooms, water, warm water, hot water or salt solution, further acid, alkaline aqueous solution, organic solvent and the like to the crushed powder, powder for 2 hours to 100 times the amount of solvent, any time, any Temperature. Further, β-glucan can be obtained by solid-liquid separation of the extract. Among these, β-glucan extracted with water, hot water or hot water is preferable, and β-glucan extracted with hot water having a temperature of 80 ° C. or lower and 4 ° C. or higher is more preferable. Further, an extraction accelerator such as an enzyme solution may be added during the extraction.
[0034]
In addition, β-glucan derived from microorganisms and basidiomycetes includes 1-2-D-glucopyranose bond, 1-3-β-D-glucopyranose bond, 1-4-β-D-glucopyranose bond, Β-glucan having at least two kinds of -6-β-D-glucopyranose linkages is preferable, and in particular, 1-3-β-D-glucopyranose linkage, β-glucan consisting of 1-4-β-D-glucopyranose linkage, Alternatively, a β-glucan consisting of a 1-3-β-D-glucopyranose bond or a 1-6-D-glucopyranose bond, or a 1-3-β-D-glucopyranose bond, 1-4-β- It preferably contains β-glucan consisting of a D-glucopyranose bond and a 1-6-β-D-glucopyranose bond.
[0035]
The β-glucan used in the present invention is particularly preferably water-soluble. In the development of dental caries, microorganisms in the oral cavity, especially enzymes and glucosyltransferases produced by Streptococcus mutans are involved, and some of the sucrose in the food and drink remaining in the oral cavity is converted into water by the action of glucosyltransferase. It has been shown to turn into insoluble and strongly adherent glucan, which, along with oral microbes, adheres to the tooth surface and forms plaque (plaque). It is considered that the β-glucan used in the present invention has an effect of suppressing the production of glucan in the oral cavity that is strongly water-insoluble and adherent.
[0036]
The microorganisms or basidiomycetes-derived β-glucan used in the present invention is a polymer, and β-glucan having any weight-average molecular weight can be used.However, since the water solubility is improved with the decrease in molecular weight, the molecular weight is 3,000,000 or less. , Preferably 500,000 or less, more preferably 100,000 or less. Β-glucan obtained from microorganisms or basidiomycetes may be reduced in molecular weight by a known method so as to improve water solubility, and β-glucan of low molecular weight may be directly extracted.
[0037]
Next, mevalonic acid will be described. The mevalonic acid referred to in the present invention is involved in the biosynthesis and metabolism of isoprenoid-related substances of an extremely large number of organisms in nature, but the lactone form is mevalonolactone, and mevalonolactone is used as mevalonic acid in an aqueous solution. Exists. In this specification, mevalonic acid and mevalonolactone are treated as synonyms.
[0038]
As a method for mass production of mevalonic acid, for example, a method of producing mevalonic acid from a microorganism is known (Japanese Patent Publication No. 7-89938, Japanese Patent Publication No. 7-89939, Japanese Patent Publication No. 7-89940, Japanese Patent Publication No. 7-89940). In this method, natural R-mevalonic acid can be obtained.
[0039]
In addition, mevalonic acid obtained by chemical synthesis has two types of optical isomers, R-mevalonic acid and S-mevalonic acid. In the present invention, these racemates can be used as they are, Alternatively, only natural R-mevalonic acid may be used after being divided. Of course, it is also possible to use only S-mevalonic acid separately.
[0040]
The mevalonic acid used in the present invention is preferably a natural R-mevalonic acid from the viewpoint of compatibility with living organisms and safety.
[0041]
In the present invention, as the mevalonic acid, a monovalent or divalent metal salt of mevalonic acid, for example, a salt of sodium, potassium, calcium or the like, or an alcohol, for example, methyl alcohol, ethyl alcohol, butyl alcohol, propylene glycol , Glycerin, isopropyl alcohol and the like. In salts, R-mevalonate and S-mevalonate are present, and in esters, R-mevalonate and S-mevalonate are present, and their racemic forms can be used as they are. From the viewpoint of safety and safety, natural R-mevalonates and R-mevalonates are preferred.
[0042]
The glucosyltransferase inhibitor, plaque formation inhibitor, gingivitis inhibitor, and periodontal disease inhibitor of the present invention contain β-glucan and mevalonic acid as active ingredients as described above. Each of them may be used alone as an active ingredient, but a combined effect can be expected when used in combination. Although the content is not particularly limited, in the case of β-glucan, 0.01 to 100% by weight is preferably based on the total amount of each of the glucosyltransferase inhibitor, the plaque formation inhibitor, the gingivitis inhibitor, and the periodontal disease inhibitor. In the case of mevalonic acid, or in the case of using β-glucan and mevalonic acid together, 0.01 to 100% by weight of the total amount of each of the glucosyltransferase inhibitor, the plaque formation inhibitor, the gingivitis inhibitor, and the periodontal disease inhibitor is used. Is preferred.
[0043]
The glucosyltransferase inhibitor according to the present invention contains, in addition to β-glucan and mevalonic acid, other known glucosyltransferase inhibitors, optional auxiliaries, excipients, water or an organic solvent for use as a solution, and the like. Can be done. Similarly, the plaque formation inhibitor, gingivitis inhibitor, and periodontal disease inhibitor of the present invention include, in addition to β-glucan and mevalonic acid, other known plaque formation inhibitors, gingivitis inhibitors, periodontal disease inhibitors , Optional auxiliaries, excipients, water or organic solvents for use as a solution, and the like. The glucosyltransferase inhibitor, plaque formation inhibitor, gingivitis inhibitor, and periodontal disease inhibitor of the present invention may be used alone or in combination of two or more.
[0044]
The glucosyltransferase inhibitor and the plaque formation inhibitor according to the present invention can be added to oral preparations and their effects can be used to prevent glucan production in the oral cavity and to suppress plaque formation. In addition, the gingivitis inhibitor and the periodontal disease inhibitor can be used for suppressing gingivitis and periodontal disease by adding to oral preparations.
[0045]
Examples of suitable oral preparations to be added include various toothpastes, mouthwashes, troches, chewing gums, oral pasta, gingival massage creams, gargles, mouth fresheners and the like.
[0046]
The addition of the glucosyltransferase inhibitor, plaque formation inhibitor, gingivitis inhibitor, periodontal disease inhibitor of the present invention does not limit other oral agent components or oral agent manufacturing methods, and other components. Examples of the components and additives include abrasives such as calcium hydrogen phosphate, calcium carbonate, insoluble sodium metaphosphate, aluminosilicate, silicic anhydride, and resin; long-chain sodium alkyl sulfate, sodium lauryl sulfoacetate, lauryl diethanol amide , Sucrose fatty acid esters, etc .; CMC, hydroxyethyl cellulose, alginate, carrageenan, gum arabic, polyvinyl alcohol, etc .; binders, such as polyethylene glycol, sorbitol, glycerin, propylene glycol; saccharin, stevioside Kind, Sweeteners such as lithilic acid, thaumatin, aspartame; preservatives such as dehydroacetic acid, sodium dehydroacetate; flavors such as menthol, carvone, eugenol, anethole, peppermint oil, spearmint oil, peppermint oil, eucalyptus oil, ginger oil, anise oil Raw materials such as various dyes ordinarily used for the production of oral agents can be arbitrarily selected according to the type and use of the product, and can be produced by a conventional method.
[0047]
In addition, the oral preparations may be used in combination with other known glucosyltransferase inhibitors, plaque formation inhibitors, gingivitis inhibitors, periodontal disease inhibitors, and are further effective antibacterial agents against Streptococcus mutans An agent may be added. By adding an optional anti-inflammatory agent, antibacterial agent, deodorant and the like, a more excellent oral agent can be provided. Examples of anti-inflammatory agents that can be added include extracts of Acacia catechu, Licorice, Ouarushi, Ougon, Kouki, Psycho, Hawthorn, Perilla, Peony, Sakuhakuhi, Kyounin, Tissou, Clove, Tonin, Nikuzuku, Buttonpi, Mulberry leaves, etc. Azulene, allantoin, ursolic acid, oleanolic acid, glycyrrhizic acid, glycyrrhetinic acid or derivatives thereof; tocopherol, tranexamic acid and the like; and examples of antibacterial agents that can be added include gobishi, saicin, sansho, and shaw. Extracts such as ginger, dill, thyme, rosemary, oil-soluble licorice extract and the like; ascorbic acid, mutastin, humic acid, linoleic acid, linolenic acid and the like. Further, examples of the deodorant that can be used in combination include extracts such as amateur, fennel, vulgar oak, calyx, pepper, mace, sage, perilla, ginkgo, oyster leaf, green tea, oolong tea, capsicum, tamarind husk; rosin, oyster Astringent, actizol, chlorophyllin derivatives, ellagic acid, chlorhexidine, Maillard reactants, and the like.
[0048]
The glucosyltransferase inhibitor, plaque formation inhibitor, gingivitis inhibitor, and periodontal disease inhibitor containing the β-glucan and / or mevalonic acid of the present invention as active ingredients are readily soluble in water at practical concentrations. Therefore, there is no particular difficulty in formulating it into an oral preparation, particularly an aqueous oral solution. The preferred addition rate varies depending on the strength of the activity and the substance to be added, but the total amount of β-glucan and mevalonic acid, which are the active ingredients, is about 0.001 to 5.0% by weight based on the total amount of the oral preparation. And a particularly preferable compounding ratio is about 0.1 to 1.0% by weight.
[0049]
The glucosyltransferase inhibitor, the plaque formation inhibitor, the gingivitis inhibitor, and the periodontal disease inhibitor according to the present invention are used not only as oral agents but also as foods and drinks, particularly when added to foods and drinks containing sucrose. It can be used as a caries preventive agent and a caries preventive food / drink used for preventing the formation of glucans and plaques in the oral cavity and preventing caries. Examples of foods and drinks to which the caries preventive agent is added include soft drinks, confectionery, bread, candy, chewing gum, gummy, jelly, chocolate, tablet confectionery, processed food, pet food, feed, and the like. Thus, these become caries-preventive foods and drinks.
[0050]
【Example】
Hereinafter, the present invention will be described with reference to examples.
[0051]
[Test Example 1] Measurement of content of β-glucan derived from grain
The analysis of β-glucan derived from grains was performed by the McCleary method (enzymatic method) using a β-glucan measurement kit from Megazyme. First, when the measurement sample was a powder, the sample was sifted through a 500 μm (30 mesh) sieve to measure the water content, 100 mg of the sample was placed in a 17 ml tube, and 200 μl of a 50% ethanol solution was added and dispersed. Next, 4 ml of 20 mM phosphate buffer (pH 6.5) was added, mixed well, and heated in a boiling water bath for 1 minute. Mix well and heat in water bath for another 2 minutes. After cooling to 50 ° C., the mixture was allowed to stand for 5 minutes, and then 200 μl (10 U) of a lichenase enzyme solution (the vial included in the kit was diluted with 20 ml of 20 mM phosphate buffer, and the remaining amount was frozen and stored) was added to each tube. The reaction was performed at 50 ° C. for 1 hour. 5 ml of 200 mM acetate buffer (pH 4.0) was added to the tube and mixed gently. The mixture was left at room temperature for 5 minutes and centrifuged to obtain a supernatant. Take 100 μl into three tubes, one with 100 μl 50 mM acetate buffer (pH 4.0) and the other two with 100 μl (0.2 U) β-glucosidase solution (20 ml vial provided with the kit). Was diluted with 50 mM acetate buffer, and the remaining amount was stored frozen), and the mixture was reacted at 50 ° C. for 10 minutes. 3 ml of a glucose oxidase / peroxidase solution was added and reacted at 50 ° C. for 20 minutes, and the absorbance (EA) at 510 nm of each sample was measured. β-glucan content was determined by the following equation.
[0052]
βglucan (%, W / W) = (EA) × (F / W) × 8.46
F = (100) / (absorbance of glucose 100 μg)
W = calculated anhydride weight (mg)
[0053]
When the measurement sample is an extract (liquid) obtained by extracting β-glucan, the content may be measured as an extract (solid or powder) as described below. That is, a two-fold amount of ethanol was added to the β-glucan extract, mixed well, and the precipitate was collected by centrifugation, thoroughly dried and pulverized to obtain a β-glucan extract (solid). After measuring the water content, the β-glucan extract was analyzed by the McCleary method (enzymatic method) using a β-glucan measurement kit from Megazyme. 50 mg of each precipitated sample was placed in a 17 ml tube, and 200 μl of a 50% ethanol solution was added and dispersed. Thereafter, the measurement was performed in the same manner as described above.
[0054]
[Test Example 2] Measurement of β-glucan content derived from microorganisms or basidiomycetes
β-glucan is analyzed by measuring the total amount of polysaccharide precipitated by alcohol by the phenol sulfate method, and then confirming and quantifying β-glucan in the precipitated polysaccharide by Seikagaku Corporation (1-3) -β This was performed using a kit for detecting and measuring β-glucan containing -D- linkage. First, the total amount of polysaccharide in the measurement sample was measured by the phenol sulfate method. That is, 30 μl of distilled water was added to 30 μl of the sample solution, 120 μl of a phosphate buffer (pH 6.9) containing 300 mM NaCl was added thereto, and 540 μl (3 times) of ethanol was further added, and the mixture was added to −15 ° C. for 10 minutes. The polysaccharide was precipitated on standing. After removing the supernatant, 100 μl of distilled water was added to dissolve. 100 μl of a 5% phenol aqueous solution and 500 μl of sulfuric acid were added thereto and reacted. The absorbance at 490 nm was measured using a blank obtained by adding a phenol solution and sulfuric acid to 100 μl of distilled water without adding any sample. A standard curve was prepared using a two-fold dilution series prepared from 10 mg / ml of pullulan as a standard sample, and the amount of polysaccharide was quantified.
[0055]
Next, a solution having a total polysaccharide content of about 1 to 0.1 mg / ml was first diluted 10-fold with 0.5 M NaOH, and then diluted with β-glucan-free distilled water. ^-10 Dilutions were prepared up to. 50 μl of the β-glucan dilution was placed in a tube, 50 μl of the main reaction reagent was added, and the mixture was incubated at 37 ° C. for 30 minutes. Subsequently, 50 μl of a sodium nitrite solution, 50 μl of ammonium sulfamate, and 50 μl of an N-methyl-2-pyrrolidone solution were added and reacted, and the absorbance of the solution was measured at 545 nm (the target wavelength: 630 nm). In addition, a calibration curve was obtained with a β-glucan solution of 7.5 to 60 pg / ml using the attached β-glucan standard, and the concentration of each β-glucan solution was calculated.
[0056]
[Test Example 3] Measurement of β-glucan molecular weight
The measurement of the molecular weight of β-glucan was as follows. That is, 5 mg of the extract was placed in a tube, 0.5 ml of distilled water was added, and the extract was dissolved in boiling water. A sample for HPLC was obtained through a 0.22 μm filter. For separation, a packed column KS-805 (manufactured by Showa Denko KK) from Shodex, which is an HPLC gel filtration column, was used at a flow rate of 0.6 ml / min. The temperature was 50 ° C., the detection was performed using an RI detector, and the separation solvent was water. The molecular weight marker was measured using Shodex pullulan standard solution P-82 (manufactured by Showa Denko KK).
[0057]
When the extracted β-glucan was an extract (liquid), a double volume of ethanol was first added, and the mixture was cooled to −20 ° C. and left for 1 hour to obtain a precipitate. 5 mg of the obtained precipitate was placed in a tube, and thereafter, the same operation as in the case of the extract was performed, and the molecular weight was measured.
[0058]
[Example of preparation of β-glucan raw material from grain and production of extraction accelerator]
Sticky bare barley was ground with a grinding mill and milled to a yield of 82%. The bran generated at this time was designated as bran-1. The barley polished to a yield of 82% was further ground by a grinding mill, and polished to a yield of 55%. The bran generated at this time was referred to as ground product-1. 20 L of tap water was added to the container (50 L), and the temperature was adjusted to 15 ° C. while stirring. 6 kg of bran-1 was added thereto, and the mixture was stirred and extracted for 2 hours. After solid-liquid separation with a continuous centrifuge, the supernatant was freeze-dried to obtain 450 g of an extraction accelerator.
[0059]
Production Example 1 [Example of extracting β-glucan from cereals]
30 L of tap water was added to a container (70 L), 150 g of the above-mentioned extraction accelerator was added with stirring, and after dissolution, 7.5 kg of the above-mentioned pulverized product-1 was added. After stirring and extracting at 50 ° C. for 2 hours, solid-liquid separation was performed using a continuous centrifuge, and then a supernatant was obtained. The obtained supernatant was boiled, and after cooling, 15 L of a slightly viscous β-glucan solution was obtained. A 2-fold amount of ethanol was added to the obtained β-glucan solution, and the precipitate was recovered and dried to obtain 460 g of β-glucan (sample 1). As a result of analysis according to Test Example 1, the purity of β-glucan was 91%. As a result of analysis according to Test Example 2, the extract was detected at a molecular weight of 200,000 to 10,000, and the maximum peak was at a molecular weight of 40,000. In addition, it was confirmed that the maximum peak was β-glucan by the method of Test Example 1.
[0060]
Production Example 2 [Production of R-mevalonic acid]
R-mevalonic acid (sample 2) was produced according to the method described in JP-B-7-89940, page 3, left column, line 11 to page 4, right column, line 4.
[0061]
Production Example 3 [Production of basidiomycete-derived β-glucan]
The fruit body of Kawariharatake was crushed and pulverized, and 50 liters of hot water was added to 10 kg of the pulverized material, and subjected to a hot water extraction treatment under boiling conditions for 3 hours with gentle stirring. After hot water extraction, the mixture was centrifuged to obtain a separated liquid. A 3-fold amount of 99% ethyl alcohol was added to the separated solution to obtain a precipitate, which was lyophilized to obtain 1200 g of β-glucan (sample 3). The amount of β-glucan contained in 1 g of Sample 3 was calculated to be 860 mg. The maximum peak molecular weight was 1,000,000.
[0062]
Production Example 4 [Production of microorganism-derived β-glucan]
One liter of 2% sodium hydroxide was added to 100 g of the cell wall (yeast cell wall E) prepared from commercially available pressed baker's yeast, and the mixture was stirred and extracted at 4 ° C. for 24 hours. The centrifuged extract was neutralized with HCl and precipitated with twice the amount of ethanol to obtain 20 g of β-glucan (sample 4). The amount of β-glucan contained in 10 mg of Sample 4 was calculated to be 4.2 mg. The maximum peak molecular weight was 1.6 million.
[0063]
[Test Example 4] Glucosyltransferase inhibition rate
Inhibition rate of glucosyltransferase activity of Sample 1 (β-glucan), Sample 2 (R-mevalonic acid), Sample 3 (β-glucan), and Sample 4 (β-glucan) obtained in Production Examples 1 to 4 by the following method Was examined. Further, a 1: 1 (weight ratio) mixture of Sample 1 and Sample 2 was used as Sample 5 and examined in the same manner.
[0064]
Glucosyltransferase inhibition rate measurement method:
1.0 ml of a 2% sucrose solution (using a 50 mM potassium phosphate buffer at pH 6.5)
0.1 ml of 1% sodium azide
Crude glucosyltransferase solution 50 μl
50 μl of sample aqueous solution (concentration 0.005 to 2 mg / ml)
0.1 mM 50 mM potassium phosphate buffer (pH 6.5)
The enzyme reaction solution obtained by mixing the above solutions is put into a test tube, and the test tube is left at 37 ° C. for 20 hours with the test tube inclined at 30 °. Thereafter, the test tube is slowly tilted to remove the reaction supernatant, and the glucan generated by the enzyme reaction and attached to the test tube is washed three times with water. Thereafter, 2 ml of water is added, and the glucan is dispersed in water by ultrasonic treatment. The absorbance at a wavelength of 550 nm is measured for the glucan dispersion and the enzyme reaction solution immediately after preparation. In addition, as a control, the same operation is performed when water is added instead of the sample aqueous solution. From the measurement results, the inhibition rate of glucosyltransferase activity is calculated by the following formula.
[0065]
Inhibition rate (%) = [1- (A1-A0) / (A3-A2)] × 100
However,
A0: Absorbance when the sample was added; before the start of the enzyme reaction
A1: Absorbance when sample is added; Glucan dispersion
A2: Control (before starting enzyme reaction)
A3: Control (Glucan dispersion)
[0066]
From the results of the above test, the sample concentration at which the inhibition rate becomes 50% is determined by interpolation and displayed as an IC50 value (the smaller the IC50 value, the stronger the enzyme activity inhibiting action).
As a result, the sample 1 was 2000 μg / ml, the sample 2 was 2400 μg / ml, the sample 3 was 2200 μg / ml, the sample 4 was 2300 μg / ml, and the sample 5 was 1700 μg / ml.
[0067]
[Test Example 5] Plaque formation inhibitory action
For the sample 1 (β-glucan), the sample 2 (R-mevalonic acid), the sample 3 (β-glucan), and the sample 4 (β-glucan) obtained in Production Examples 1 to 4, the plaque formation inhibitory effect was examined by the following method. Was. A 1: 1 (weight ratio) mixture of Sample 1 and Sample 2 was similarly examined as Sample 5.
[0068]
Test of plaque formation inhibitory action: 5.35 ml of Brain Heart Infusion Broth (manufactured by Nissui Pharmaceutical) containing 2% of sucrose is added to a test tube weighed in advance. After heat sterilization, 0.15 ml of a sample solution and 0.5 ml of a culture solution of Streptococcus mutans 6715 are added, and the cells are cultured at 37 ° C. for 20 minutes. After completion of the culture, the supernatant is gently removed, and the plaque-like deposit on the tube wall of the test tube is washed with distilled water three times as it is, and then dried at 105 ° C. for 5 hours. Finally, each test tube is weighed to determine the dry weight w of the plaque-like deposit in the tube. Separately, as a blank test, the same operation as described above is performed using the solvent of the sample solution instead of the sample solution, and the dry weight W of the plaque-like deposit is obtained. From the weights w and W of the plaque-like deposits measured, the plaque formation inhibition rate is calculated by the following equation.
[0069]
Plaque formation inhibition rate (%) = (1−w / W) × 100
[0070]
The above-mentioned measurement is performed by changing the concentration of the sample solution in a stepwise manner, and the concentration IC50 of the sample solution at which the suppression rate becomes 50% is obtained by the interpolation method.
As a result, sample 1 was 400 µg / ml, sample 2 was 630 µg / ml, sample 3 was 470 µg / ml, sample 4 was 500 µg / ml, and sample 5 was 320 µg / ml.
[0071]
Hereinafter, Examples in which the oral composition of the present invention is blended will be described. The following examples were all excellent in the periodontal disease preventing effect and the effect of improving the trouble due to the periodontal disease, and the safety was also good.
[0072]
Example 1
The following mixture was tableted with a tableting machine to produce an oral preparation for preventing dental caries.
[Formulation]
Sample 1 (β glucan) 120 parts by weight
172 parts by weight of raffinose
Glycerin fatty acid ester 8 parts by weight
[0073]
Example 2
The following mixture was tableted with a tableting machine to produce an oral preparation for preventing dental caries.
[Formulation]
Sample 1 (β glucan) 100 parts by weight
Sample 2 (R-mevalonic acid) 20 parts by weight
172 parts by weight of raffinose
Glycerin fatty acid ester 8 parts by weight
[0074]
Example 3
The following mixture was formed into granules by a fluidized bed granulator to produce an instant tea having a caries preventing action.
[Formulation]
Sample 1 (β glucan) 300 parts by weight
Raffinose 290 parts by weight
100 parts by weight of oolong tea water extract
810 parts by weight of indigestible dextrin
[0075]
Example 4
1 kg of flour, 100 g of corn starch, 250 g of sugar, 125 g of margarine, 5 g of salt, 25 g of sodium carbonate, 9 g of ammonium carbonate, 6 g of lecithin, 75 g of whole eggs, 50 g of calcium lactate, 2 g of sample 1 (β-glucan), and 350 g of water are kneaded and mixed. Was prepared, and then spread, molded, and roasted by a conventional method to produce a biscuit in which caries prevention measures were taken.
[0076]
Example 5
330 g of isomerized sugar, 140 g of sugar and 270 g of water are mixed and heated, and a solution obtained by dissolving 80 g of gelatin, 10 g of sample 1 (β-glucan) in 150 g of water, and 8 g of concentrated lemon juice are poured into a mold. By cooling, a caries-preventive gummy candy was produced.
[0077]
Example 6
25 parts by weight of gum base and 14 parts by weight of starch syrup are mixed in a chewing gum trial mixer, and then 35 parts by weight of sugar, 25 parts by weight of fructose, 14 parts by weight of sample 1 (β-glucan), stevia sweetener / marmilon (Maruzen Pharmaceutical Co., Ltd.) (Company product) 0.4 parts by weight of the mixture was added in several portions and kneaded well. Next, 1 part by weight of glycerin was added, mixed well, taken out of the mixer, and rolled with a roller to produce a chewing gum having a caries-preventing action.
[0078]
Example 7
Mouthwash A having the following composition was produced.
[Formulation]
15 parts by weight of ethanol
Glycerin 10 parts by weight
0.05 parts by weight of ascorbic acid
0.2 parts by weight of sodium citrate
0.2 parts by weight of sodium benzoate
0.2 parts by weight of sodium lauryl sulfate
Saccharin sodium 0.05 parts by weight
Sample 1 (β glucan) 0.2 parts by weight
1-menthol 0.05 parts by weight
Purified water balance
100 parts by weight in total
[0079]
Example 8
Mouthwash B having the following composition was produced.
[Formulation]
15 parts by weight of ethanol
Glycerin 10 parts by weight
0.05 parts by weight of ascorbic acid
0.2 parts by weight of sodium citrate
0.2 parts by weight of sodium benzoate
0.2 parts by weight of sodium lauryl sulfate
Saccharin sodium 0.05 parts by weight
Sample 2 (R-mevalonic acid) 0.2 parts by weight
1-menthol 0.05 parts by weight
Purified water balance
100 parts by weight in total
[0080]
Example 9
Mouthwash C having the following composition was produced.
[Formulation]
15 parts by weight of ethanol
Glycerin 10 parts by weight
0.05 parts by weight of ascorbic acid
0.2 parts by weight of sodium citrate
0.2 parts by weight of sodium benzoate
0.2 parts by weight of sodium lauryl sulfate
Saccharin sodium 0.05 parts by weight
Sample 1 (β glucan) 0.1 parts by weight
Sample 2 (R-mevalonic acid) 0.1 parts by weight
1-menthol 0.05 parts by weight
Purified water balance
100 parts by weight in total
[0081]
Example 10
A toothpaste A having the following composition was produced.
[Formulation]
Silicic anhydride (average particle size = 10 μm) 20.0% by weight
Sodium polyacrylate (degree of polymerization = 15000) 0.5% by weight
Xanthan gum (Mw = 100-1.5 million) 0.5% by weight
Propylene glycol 5.0% by weight
70% sorbite liquid 20.0% by weight
Saccharin sodium 0.1% by weight
Sodium benzoate 0.3% by weight
Sodium lauryl sulfate 1.5% by weight
Sample 1 (β glucan) 0.5% by weight
Spice 1.0% by weight
Purified water residue
100.0% by weight in total
[0082]
Example 11
A toothpaste B having the following composition was produced.
[Formulation]
Silicic anhydride (average particle size = 10 μm) 20.0% by weight
Sodium polyacrylate (degree of polymerization = 15000) 0.5% by weight
Xanthan gum (Mw = 100-1.5 million) 0.5% by weight
Propylene glycol 5.0% by weight
70% sorbite liquid 20.0% by weight
Saccharin sodium 0.1% by weight
Sodium benzoate 0.3% by weight
Sodium lauryl sulfate 1.5% by weight
Sample 2 (R-mevalonic acid) 0.5% by weight
Spice 1.0% by weight
Purified water residue
100.0% by weight in total
[0083]
Example 12
A toothpaste C having the following composition was produced.
[Formulation]
Silicic anhydride (average particle size = 10 μm) 20.0% by weight
Sodium polyacrylate (degree of polymerization = 15000) 0.5% by weight
Xanthan gum (Mw = 100-1.5 million) 0.5% by weight
Propylene glycol 5.0% by weight
70% sorbite liquid 20.0% by weight
Saccharin sodium 0.1% by weight
Sodium benzoate 0.3% by weight
Sodium lauryl sulfate 1.5% by weight
Sample 1 (β glucan) 0.3% by weight
Sample 2 (R-mevalonic acid) 0.2% by weight
Spice 1.0% by weight
Purified water residue
100.0% by weight in total
[0084]
Example 13
A toothpaste D having the following composition was produced.
[Formulation]
Calcium hydrogen phosphate dihydrate (average particle size = 10 μm) 45.0% by weight
Silicic anhydride (average particle size = 10 μm) 2.0% by weight
Sodium carboxymethylcellulose
Substitution degree: 0.7, Mw = 300,000) 0.3% by weight
Sodium carboxymethylcellulose
(Substitution degree: 1.0, Mw = 300,000) 0.7% by weight
Carrageenan (Mw = 500,000) 0.1% by weight
Propylene glycol 3.0% by weight
60% sorbite liquid 25.0% by weight
0.2% by weight methyl paraoxybenzoate
Sodium benzoate 0.5% by weight
Saccharin sodium 0.2% by weight
0.73% by weight of sodium monofluorophosphate
Tranexamic acid 0.05% by weight
Isopropyl methyl phenol 0.1% by weight
Sodium lauryl sulfate 1.2% by weight
Lauroyl sarcosine sodium 0.3% by weight
Sample 1 (β glucan) 0.2% by weight
Spice 1.0% by weight
Purified water residue
100.0% by weight in total
[0085]
Example 14
A liquid dentifrice A having the following composition was produced.
[Formulation]
Precipitable silica (average particle size = 10 μm) 20.0% by weight
Butyl paraoxybenzoate 0.01% by weight
Xanthan gum (Mw = 100 to 1.5 million) 0.2% by weight
Sodium polyacrylate (Mw = 15000) 0.15% by weight
Propylene glycol 2.0% by weight
Sorbit 35.0% by weight
Glycerin 25.0% by weight
Saccharin sodium 0.1% by weight
Spice 1.0% by weight
Dye (Brilliant Blue) 0.001% by weight Sodium lauryl sulfate 1.5% by weight
Decaglyceryl monolaurate 2.0% by weight
Sample 1 (β glucan) 0.1% by weight
0.25% by weight of fragrance
Purified water residue
100.0% by weight in total
[0086]
Example 15
A liquid dentifrice B having the following composition was produced.
[Formulation]
Precipitable silica (average particle size = 10 μm) 20.0% by weight
Butyl paraoxybenzoate 0.01% by weight
Xanthan gum (Mw = 100 to 1.5 million) 0.2% by weight
Sodium polyacrylate (Mw = 15000) 0.15% by weight
Propylene glycol 2.0% by weight
Sorbit 35.0% by weight
Glycerin 25.0% by weight
Saccharin sodium 0.1% by weight
Spice 1.0% by weight
Dye (Brilliant Blue) 0.001% by weight Sodium lauryl sulfate 1.5% by weight
Decaglyceryl monolaurate 2.0% by weight
Sample 2 (R-mevalonic acid) 0.1% by weight
0.25% by weight of fragrance
Purified water residue
100.0% by weight in total
[0087]
Example 16
A liquid dentifrice C having the following composition was produced.
[Formulation]
Precipitable silica (average particle size = 10 μm) 20.0% by weight
Butyl paraoxybenzoate 0.01% by weight
Xanthan gum (Mw = 100 to 1.5 million) 0.2% by weight
Sodium polyacrylate (Mw = 15000) 0.15% by weight
Propylene glycol 2.0% by weight
Sorbit 35.0% by weight
Glycerin 25.0% by weight
Saccharin sodium 0.1% by weight
Spice 1.0% by weight
Dye (Brilliant Blue) 0.001% by weight Sodium lauryl sulfate 1.5% by weight
Decaglyceryl monolaurate 2.0% by weight
Sample 1 (β glucan) 0.07% by weight
Sample 2 (R-mevalonic acid) 0.03% by weight
0.25% by weight of fragrance
Purified water residue
100.0% by weight in total
[0088]
Example 17
A liquid dentifrice D having the following composition was produced.
[Formulation]
Aluminum hydroxide 25.0% by weight
Sodium polyacrylate (degree of polymerization = 15000) 0.1% by weight
Carboxymethylcellulose sodium (Mw = 300,000) 0.2% by weight
Propylene glycol 2.0% by weight
Sorbit 15.0% by weight
Glycerin 40.0% by weight
Saccharin sodium 0.1% by weight
Spice 1.0% by weight
Sodium lauryl sulfate 1.5% by weight
1.0% by weight of decaglyceryl monolaurate
Sample 1 (β glucan) 0.3% by weight
0.25% by weight of fragrance
Purified water residue
100.0% by weight in total
[0089]
Example 18
An oral freshener having the following composition was produced.
[Formulation]
Ethanol 50.0% by weight
Glycerin 15.0% by weight
P. O. E (60) hydrogenated castor oil 3.0% by weight
1-menthol 1.0% by weight
Sample 1 (β glucan) 0.2% by weight
Sample 2 (R-mevalonic acid) 0.1% by weight
0.3% by weight of fragrance
Purified water residue
100.0% by weight in total
[0090]
Example 19
An oral pasta having the following composition was produced.
[Formulation]
Cetanol 10.0% by weight
Squalane 20.0% by weight
Precipitable silica 5.0% by weight
P. O. E (40) hydrogenated castor oil 0.1% by weight
Sorbitan monooleate 1.0% by weight
Sodium lauryl sulfate 0.2% by weight
Glycyrrhetinic acid 0.1% by weight
0.6% by weight of saccharin sodium
Sample 1 (β glucan) 0.3% by weight
Sample 2 (R-mevalonic acid) 0.2% by weight
0.5% by weight of ε-aminocaproic acid
Ethyl salicylate 0.2% by weight
Isoeugenol 0.1% by weight
Mentone 0.05% by weight
0.6% by weight of fragrance
Purified water residue
100.0% by weight in total
[0091]
Example 20
An oral troche A having the following composition was produced.
[Formulation]
Lactose 97.0% by weight
P. O. E (60) Monostearate 0.2% by weight
Sodium lauryl sulfate 0.05% by weight
Chlorhexidine gluconate 0.02% by weight
Stevia extract 0.2% by weight
Sample 1 (β glucan) 0.3% by weight
0.2% by weight of methyl salicylate
Isoeugenol 0.05% by weight
Spice 0.02% by weight
Hydroxyethyl cellulose (Mw = 1,000,000) remaining
100.0% by weight in total
[0092]
Example 21
An oral troche B having the following composition was produced.
[Formulation]
Lactose 97.0% by weight
P. O. E (60) Monostearate 0.2% by weight
Sodium lauryl sulfate 0.05% by weight
Chlorhexidine gluconate 0.02% by weight
Stevia extract 0.2% by weight
Sample 2 (R-mevalonic acid) 0.3% by weight
0.2% by weight of methyl salicylate
Isoeugenol 0.05% by weight
Spice 0.02% by weight
Hydroxyethyl cellulose (Mw = 1,000,000) remaining
100.0% by weight in total
[0093]
Example 22
An oral troche C having the following composition was produced.
[Formulation]
Lactose 97.0% by weight
P. O. E (60) Monostearate 0.2% by weight
Sodium lauryl sulfate 0.05% by weight
Chlorhexidine gluconate 0.02% by weight
Stevia extract 0.2% by weight
Sample 1 (β glucan) 0.15% by weight
Sample 2 (R-mevalonic acid) 0.15% by weight
0.2% by weight of methyl salicylate
Isoeugenol 0.05% by weight
Spice 0.02% by weight
Hydroxyethyl cellulose (Mw = 1,000,000) remaining
100.0% by weight in total
[0094]
(Clinical trial)
With the toothpastes of Examples 10 to 13 above, five subjects each were allowed to perform toothbrushing under the following conditions for a certain period of time. Specifically, after each meal three times a day, toothbrushing was performed for one month. Before and after this test, the effects were evaluated for the following test items.
[0095]
[Test 1] Evaluation of caries-related factors
(1) Measurement of salivary secretion
The measurement of the salivary secretion of the subject was performed as follows. First, the subject was sat upright, given paraffin and chewed. After 30 seconds, saliva was swallowed once, and immediately after that, saliva was exhaled through a funnel into a graduated test tube while chewing for 5 minutes continuously, and the amount of saliva secreted for 5 minutes was measured. Table 1 shows the results. For salivary secretion, ++ indicates that an increase of 30% or more was observed, + indicates that an increase of 5% to less than 30% was observed, 0 indicates an increase or decrease within ± 5%, and-indicates a change of 5% or less. It means that it decreased more than%.
[0096]
[Table 1]
Table 1

[0097]
(2) Measurement of salivary buffer capacity
The measurement was performed using Dentobuff Strip (manufactured by Morimura Co., Ltd.), which can measure salivary buffer capacity. According to Dentobuff Strip, salivary buffer capacity can be evaluated in four steps. Specifically, first, a saliva sample was taken with a pipette, dropped on a strip, and after 5 minutes, the color of the strip and the color chart were compared. In this kit, the buffer capacity of saliva is classified into immediate blue, blue, green, and yellow levels in descending order. Table 2 shows the results.
[0098]
[Table 2]
Table 2

[0099]
(3) Measurement of the number of mutans streptococci
The number of mutans streptococci in the oral cavity was determined using a Dentocult-SM kit (manufactured by Morimura Co., Ltd.). Here, Dentocult-SM is a test medium capable of examining the number of mutans streptococci in saliva. First, the strip rotated on the tongue was placed in the culture solution containing the bacitracin tablet in the kit, and cultured at 37 ° C. for 2 days. Mutans streptococci appears as blue colonies on the strip, and the density of the colonies on the strip is compared with the model chart to determine the number of bacteria in 1 ml of saliva. Was determined as ○, 500,000 as Δ, and 1,000,000 as ×.
[0100]
[Table 3]
Table 3

[0101]
(4) Measurement of bacterial count of Lactobacillus
The number of bacteria of Lactobacillus in the oral cavity was measured using a Dentocult-LB kit (manufactured by Morimura Corporation). Here, Dentocult-LB is a test medium capable of examining the bacterial count of Lactobacillus in saliva. First, a saliva sample was flowed on both sides of a slide in which a medium was impregnated in the kit, and cultured at 37 ° C. for 4 days. Lactobacillus appeared as white colonies on the slide, and the number of bacteria was determined by comparing the density of the colonies on the slide with the model chart.
[0102]
Table 4 shows the results. In the table, the number of bacteria in 1 ml of saliva is about 1000 (shown by ◎), 10,000 (shown by ○), 100,000 (shown by Δ), and 1,000,000 (shown by ×), respectively. Means
[0103]
[Table 4]
Table 4

[0104]
[Test 2]: Measurement of oral bacteria Candida
The number of Candida bacteria in the oral cavity was examined using Oricult-N (manufactured by Morimura Co., Ltd.). Saliva samples collected on both sides of the slide to which the Nickerson medium was applied were flowed and cultured at 37 ° C. for 2 days. Candida appeared as brown colonies on the slide, and the number of bacteria was determined by comparing the density of the colonies on the slide with the model chart.
[0105]
Table 5 shows the results. In the table, the number of bacteria in 1 ml of saliva is about 1000 (shown by ◎), 10,000 (shown by ○), 100,000 (shown by Δ), and 1,000,000 (shown by ×), respectively. Means
[0106]
[Table 5]
Table 5

[0107]
[Test 3]: Evaluation of periodontal disease-related factors
Porphyromonas gingivalis, Treponema denticola, and Bacteroides forsythus, which are said to be the main pathogens of adult periodontitis, were examined by a periodontopathogen test agent (Periocheck, manufactured by Sunstar Co., Ltd.) that specifically detects the enzymes produced by them. . First, a paper point inserted into the deepest part of the periodontal pocket for 30 seconds was directly charged into the vial containing the substrate and the chromogen, and the enzyme solution was added. After incubation at 37 ° C. for 15 minutes, the degree of blue change was compared with a standard color tone (color sample) to measure the peptidase activity from these periodontopathogens.
[0108]
Table 6 shows the results. In the table, (-) means negative, (+) means positive, and (++) means strong positive.
[0109]
[Table 6]
Table 6

[0110]
【The invention's effect】
According to the present invention, a glucosyltransferase inhibitor, a plaque formation inhibitor, a gingivitis and / or periodontal disease inhibitor having excellent performance, and an oral agent to which a caries-preventing action is imparted by adding them. , A caries preventive, food and drink, and the like.

Claims (6)

A glucosyltransferase inhibitor comprising β-glucan and / or mevalonic acid as an active ingredient. A plaque formation inhibitor comprising β-glucan and / or mevalonic acid as an active ingredient. A gingivitis inhibitor and / or periodontal disease inhibitor comprising β-glucan and / or mevalonic acid as an active ingredient. The glucosyltransferase inhibitor, plaque formation inhibitor, gingival salt inhibitor and / or periodontal disease inhibitor according to any one of claims 1 to 3, wherein the β-glucan is derived from a grain, a microorganism, and / or a basidiomycete. An oral agent comprising one or more selected from the group consisting of the glucosyltransferase inhibitor, the plaque formation inhibitor, the gingivitis inhibitor, and the periodontal disease inhibitor according to claims 1 to 4. A caries preventive agent comprising one or more members selected from the group consisting of the glucosyltransferase inhibitor, the plaque formation inhibitor, the gingivitis inhibitor, and the periodontal disease inhibitor according to claims 1 to 4. Or caries-preventing food or drink.