JP4623791B2 - Caries inhibitor, oral preparation and food and drink - Google Patents

Description

【００３５】
実施例２
実施例１で試験した試料について、さらに下記の方法でプラーク形成抑制作用を調べた。
【００３６】
プラーク形成抑制作用の試験：あらかじめ秤量した試験管にショ糖４％を含むブレインハートインフュージョンブロス（日水製薬製）５．３５ｍｌを加える。加熱滅菌処理後、試料溶液０．１５ｍｌおよびストレプトコッカス・ミュータンス６７１５の培養液０．５ｍｌを添加し、３７℃で２０時間培養を行う。培養終了後、上清を静かに除き、試験管管壁のプラーク状付着物をそのまま蒸留水で３回洗浄した後１０５℃で５時間乾燥する。最後に試験管ごとに秤量して、管内のプラーク状付着物の乾燥重量ｗを求める。別に、空試験として、試料溶液の代わりに試料溶液の溶媒を用いて上記と同様の操作を行い、プラーク状付着物の乾燥重量Ｗを求める。測定されたプラーク状付着物の重量ｗおよびＷより、次式によりプラーク形成抑制率を算出する。
【００３７】
プラーク形成抑制率（％）＝（１−ｗ／Ｗ）×１００
試料溶液の濃度を段階的に変更して上記測定を行い、抑制率が５０％になる試料溶液の濃度ＩＣ_５０を内挿法により求める。
その結果は表２のとおりであった。
【００３８】
【表２】
プラーク形成抑制作用

【００３９】
実施例３
下記の混合物を打錠機で打錠して、う蝕予防用健康食品を製造した。
Ｎ−バニリルパルミチン酸アミド ０．１重量部
オリーブ油 ５重量部
ラフィノース １７２重量部
デキストリン １５重量部
グリセリン脂肪酸エステル ８重量部
【００４０】
実施例４
下記の原料を混合して、う蝕防止性の練り歯磨きを製造した。
Ｎ−バニリルステアリン酸アミド ０．１重量部
オリーブ油 １重量部
第二リン酸カルシウム ４５重量部
ＣＭＣ・ナトリウム塩 １重量部
グリセリン ２０重量部
ラウリル硫酸ナトリウム ２重量部
１−メントール １重量部
水 ３０重量部
【００４１】
実施例５
下記の原料をチューインガム製造の常法により処理して、う蝕予防作用を有するチューインガムを製造した。
Ｎ−バニリルオレイン酸アミド ０．１重量部
チューインガムベース ２０重量部
ショ糖 ５０重量部
水飴 ２０重量部
軟化剤 ４重量部
香料 １重量部
【００４２】
実施例６
下記の原料を飴製造の常法に従って混合、濃縮、成形して、う蝕予防作用を有する飴を製造した。
無辛味カプサイシン類縁体混合物※ １重量部
ショ糖 ７０重量部
水飴 ３０重量部
クエン酸 １重量部
香料 ０．１重量部
水 １５重量部
（※ カプサイシンとオリーブ油をリパーゼの存在下に反応させて得られたもので、未反応オリーブ油中にＮ−バニリルオレイン酸アミド４１重量％、Ｎ−バニリルリノール酸アミド７重量％を含有する。）
【００４３】
実施例７
小麦粉１ｋｇ、コーンスターチ１００ｇ、砂糖２５０ｇ、マーガリン１２５ｇ、食塩５ｇ、炭酸ソーダ２５ｇ、炭酸アンモニウム９ｇ、レシチン６ｇ、全卵７５ｇ、乳酸カルシウム５０ｇ、無辛味カプサイシン類縁体混合物２ｇ、および水３５０ｇを混練してドウを調製し、以後、常法により延展、成形、焙焼を行なって、虫歯予防対策がなされたビスケットを製造した。なお、配合した無辛味カプサイシン類縁体混合物はカプサイシンとコーン油をリパーゼの存在下に反応させて得られたもので、未反応オリーブ油中にＮ−バニリルオレイン酸アミド１８重量％、Ｎ−バニリルリノール酸アミド３９重量％を含有する。
【００４４】
【発明の効果】
無辛味カプサイシン類縁体を利用する本発明によれば、無辛味カプサイシン類縁体がグルコシルトランスフェラーゼ阻害作用とプラーク形成抑制作用を兼備することにより優れたう蝕防止効果を達成することができる。
【００４５】
また、本発明によりう蝕予防作用が付与された飲食物は、無辛味カプサイシン類縁体の食欲増進作用、血管拡張・収縮作用、エネルギー代謝亢進作用等の有用作用も併有し、健康増進にきわめて有意義なものとなる。[0001]
[Industrial application fields]
The present invention is a glucosyltransferase inhibitor having an action of inhibiting the activity of an enzyme / glucosyltransferase that synthesizes glucan that causes caries, and an action of suppressing the formation of plaque (plaque) that is a place where oral microorganisms grow. The present invention relates to a plaque formation inhibitor, and an oral preparation and food and drink provided with a caries preventive action.
[0002]
In addition, in this specification, it is called food and drink in the meaning which includes not only the thing which a person eats and drinks but the livestock or pet animal's feed or food which may cause a caries.
[0003]
[Prior art]
The occurrence of caries involves the microorganisms in the oral cavity, especially the enzyme glucosyltransferase produced by Streptococcus mutans. That is, a part of sucrose in food and drink remaining in the oral cavity is changed into water-insoluble and highly adherent glucan by the action of glucosyltransferase, and it adheres to the tooth surface together with oral microorganisms and becomes plaque (dental Form). Then, the microorganisms in the plaque metabolize sugar in food to produce an acid, and this acid decalcifies and erodes the tooth enamel.
[0004]
Therefore, in order to prevent dental caries, not only remove the plaque attached to the tooth surface by brushing teeth, but also prevent the production of glucan by inhibiting the growth of Streptococcus mutans and the action of glucosyltransferase in the oral cavity. However, it is most effective to prevent the occurrence of plaque.
[0005]
From such a viewpoint, in recent years, oral preparations and foods and drinks that have been given a caries preventive action by containing a substance having a glucosyltransferase inhibitory action or a substance that suppresses plaque formation have been provided. Examples of glucosyltransferase inhibitors conventionally known as suitable for such applications include mutastein, herbal tannins, ellagic acid, green tea polyphenols, oolong tea extract, and the like.
[0006]
[Problems to be solved by the invention]
An object of the present invention is to provide a novel glucosyltransferase inhibitor, a plaque formation inhibitor, and oral preparations and foods and drinks to which caries preventive action is added by adding them.
[0007]
[Means for Solving the Problems]
Formulations and foods and drinks effective for preventing dental caries provided by the present invention are as follows.
[0008]
(1) A glucosyltransferase inhibitor comprising N-vanillyl fatty acid amide having an acyl group having 12 to 22 carbon atoms as an active ingredient.
[0009]
(2) A plaque formation inhibitor comprising N-vanillyl fatty acid amide having an acyl group having 12 to 22 carbon atoms as an active ingredient.
[0010]
(3) An oral preparation characterized by containing the glucosyltransferase inhibitor (1) above or (and) the plaque formation inhibitor (2) above.
[0011]
(4) A caries preventive food or drink comprising the glucosyltransferase inhibitor (1) above or (and) the plaque formation inhibitor (2) above.
[0012]
N-vanillyl fatty acid amide (3-CH ₃ O-4-OH—C ₆ H ₃ —CH ₂ —NHR; adopted in each of the inventions (1) and (2) as a main agent; Is an analog that differs in the acyl group part of capsaicin (8-methyl-N-vanillyl-6-nonenamide), which is the main pungent component of chili, but is not as hard as capsaicin, and in particular acyl A group having 13 or more carbon atoms is hardly spicy (hereinafter, the N-vanillyl fatty acid amide having an acyl group having 12 to 22 carbon atoms is referred to as a spicy capsaicin analog).
[0013]
Capsaicin has various useful effects on the body, such as increased appetite, vasodilation / contraction, increased saliva secretion, increased gastric acid secretion, increased intestinal peristalsis, decreased circulatory cholesterol, increased energy metabolism, and increased release of bioactive peptides. However, similar effects are being confirmed for the non-spicy capsaicin analogs, and the above-mentioned useful effects due to oral ingestion, which was difficult with capsaicin, have been taken advantage of the feature that there is no strong pungency like capsaicin. Utilization is under consideration.
[0014]
However, it has never been known at all that capsaicin analogs are effective in preventing caries.
[0015]
The glucosyltransferase inhibitor according to the present invention utilizes the glucosyltransferase inhibitory action first confirmed for a spicy capsaicin analog, and sucrose is glucan by the action of glucosyltransferase produced by Streptococcus mutans in the oral cavity. It is useful for preventing the change to a plaque component.
[0016]
In addition, the plaque formation inhibitor according to the present invention suppresses the glucan produced in the oral cavity through the process as described above from adhering to the teeth to form plaques, thereby forming the growth site of caries-causing microorganisms. It takes advantage of the first confirmed action of a spicy capsaicin analog that prevents it from being done.
[0017]
The oral preparation and food and drink according to the present invention are those obtained by blending the above-described glucosyltransferase inhibitor or (and) plaque formation inhibitor according to the present invention to give a strong caries-preventing action.
[0018]
Capsaicin has neither a glucosyltransferase inhibitory effect nor a plaque formation inhibitory effect, and even if it exists, there is no way to eliminate the strong pungency of capsaicin, so it is almost impossible to use capsaicin for caries prevention.
[0019]
DETAILED DESCRIPTION OF THE INVENTION
Unlike capsaicin, the spicy capsaicin analog used in the present invention is a substance that does not exist in nature. For example, it can be synthesized by the following method.
[0020]
(1) An aqueous solution of vanillylamine is brought into contact with a fatty acid having 12 to 22 carbon atoms or an ester thereof in the presence of lipase to react the vanillylamine with the fatty acid or the ester thereof.
[0021]
(2) A fatty acid having 12 to 22 carbon atoms or an ester thereof is reacted with capsaicin in the presence of lipase.
[0022]
(3) An edible fat is reacted with capsaicin in the presence of lipase (According to this production method, fat and oil containing a non-savory capsaicin analog is obtained and the fat and fat are used in applications where the coexistence of fat and oil is allowed. Mixtures with capsaicin analogs can be used as is).
[0023]
A method for producing a non-spicy capsaicin analog is described in detail, for example, in the specifications (not disclosed) of Japanese Patent Application Laid-Open No. 11-206396 and Japanese Patent Application No. 10-201078.
[0024]
Among the non-spicy capsaicin analogs, N-vanillyl oleamide and N-vanillyl are particularly preferred for the present invention because they are excellent in glucosyltransferase inhibitory action and plaque formation inhibitory action and have little pungency. Linoleic acid amide, N-vanillyl myristic acid amide, N-vanillyl palmitic acid amide, N-vanillyl stearic acid amide and the like.
[0025]
The glucosyltransferase inhibitor and the plaque formation inhibitor according to the present invention can be added to an oral preparation and the action thereof can be used for preventing glucan production in the oral cavity and suppressing plaque formation. Examples of oral preparations suitable for addition include various toothpastes, mouthwashes, troches, chewing gums, oral pasta, gingival massage creams, gargles, mouth fresheners, and the like. The preferred compounding ratio in these applications is about 0.001 to 2.0% by weight as the spicy capsaicin analog, although it varies depending on the type of spicy capsaicin analog and the type of oral preparation.
[0026]
The addition of the glucosyltransferase inhibitor and the plaque formation inhibitor according to the present invention does not limit other oral preparation components and oral preparation methods, such as calcium hydrogen phosphate, calcium carbonate, insoluble sodium metaphosphate, Polishing agents such as aluminosilicate, silicic anhydride, resin, etc .; surfactants such as long-chain sodium alkyl sulfate, sodium lauryl sulfoacetate, lauryl diethanolamide, sucrose fatty acid ester; CMC, hydroxyethyl cellulose, alginate, carrageenan, gum arabic , Polyvinyl alcohol and other binders; polyethylene glycol, sorbitol, glycerin, propylene glycol and other viscous agents; saccharin, steviosides, glycyrrhizic acid, thaumatin, aspartame and other sweeteners; Preservatives such as roacetic acid and sodium dehydroacetate; menthol, carvone, eugenol, anethole, peppermint oil, spearmint oil, peppermint oil, eucalyptus oil, ginger oil, anise oil, etc .; The raw material to be used can be arbitrarily selected according to the type and use of the product, and can be produced by a conventional method.
[0027]
Moreover, when producing an oral preparation, other glucosyltransferase inhibitors may be used in combination, and an antibacterial agent effective against Streptococcus mutans may be added. By adding an optional anti-inflammatory agent, antibacterial agent, deodorant and the like, a better oral preparation can be provided. Examples of anti-inflammatory agents that can be added include extracts of Acacia yak, licorice, waurusushi, cormorant, cormorant, psycho, hawthorn, perilla, peonies, perch, pylon, tiso, clove, tonin, nutmoku, buttonpi, mulberry leaves, etc. Azulene, allantoin, ursolic acid, oleanolic acid, glycyrrhizic acid, glycyrrhetinic acid or a derivative thereof; Examples include extracts such as kyou, dill, thyme, rosemary, oil-soluble licorice extract; ascorbic acid, mutastine, humic acid, linoleic acid, linolenic acid and the like. Furthermore, examples of deodorants that can be used in combination include extracts such as amateur, fennel, radish, keihi, pepper, mace, sage, perilla, ginkgo, oyster leaf, green tea, oolong tea, red pepper, tamarind husk, etc .; rosin, oyster Astringents, actinols, chlorophyllin derivatives, ellagic acid, chlorhexidine, Maillard reactants and the like can be mentioned.
[0028]
The glucosyltransferase inhibitor and the plaque formation inhibitor according to the present invention are added not only to oral preparations but also to foods and drinks, particularly foods and drinks containing sucrose, to prevent the formation of glucan and plaque in the oral cavity. It can be used to prevent erosion. Examples of suitable foods and drinks to be added include soft drinks, confectionery, bread, candy, chewing gum, gummy, jelly, chocolate, tablet confectionery, pet food, and the like. Although the suitable mixture rate in these uses also changes with kinds etc. of food-and-drink to be added, it is about 0.001-2.0 weight% normally as a non-spicy capsaicin analog.
[0029]
Spicy capsaicin analogs are not very soluble in water, but are well soluble in fats and alcohols. Accordingly, when formulated as a glucosyltransferase inhibitor or a plaque formation inhibitor, or when added to oral preparations, foods and drinks, etc., those dissolved in oils or alcohols are used as necessary.
[0030]
【Example】
Hereinafter, the present invention will be described with reference to examples.
Example 1
N-vanillyl myristic acid amide, N-vanillyl palmitic acid amide, N-vanillyl stearic acid amide, and capsaicin were examined for inhibition rate of glucosyltransferase activity by the following method.
[0031]
Glucosyltransferase inhibition rate measurement method:
1.0 ml of 4% sucrose solution (using 50 mM potassium phosphate buffer at pH 6.5)
0.1% sodium azide 0.1ml
Crude glucosyltransferase solution 50 μl
Sample solution (solvent: dimethyl sulfoxide; concentration: 0.05 to 20 mg / ml) 50 μl
50 mM potassium phosphate buffer (pH 6.5) 0.1 ml
[0032]
The enzyme reaction solution obtained by mixing the above solutions is put in a test tube and allowed to stand at 37 ° C. for 5 hours. Thereafter, the test tube is slowly tilted to remove the reaction supernatant, and the glucan produced 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 sonication. The absorbance at a wavelength of 550 nm is measured for the glucan dispersion and the enzyme reaction solution immediately after preparation. 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.
Inhibition rate (%) = [1- (A ₁ −A ₀ ) / (A ₃ −A ₂ )] × 100
However, A ₀ : Absorbance when the sample is added; A ₁ before the enzyme reaction is started A ₁ : Absorbance when the sample is added; Glucan dispersion A ₂ : Control (before the enzyme reaction is started)
A ₃ : Control (glucan dispersion)
[0033]
Determined by interpolation of the sample concentration at which the inhibition ratio becomes 50% from the results of the test, and displays it as an IC ₅₀ value (strong enzyme inhibitory activity as an IC ₅₀ value is small).
The results are shown in Table 1.
[0034]
[Table 1]
Glucosyltransferase inhibitory action

[0035]
Example 2
About the sample tested in Example 1, the plaque formation inhibitory effect was further investigated by the following method.
[0036]
Test for inhibiting plaque formation: 5.35 ml of Brain Heart Infusion Broth (Nissui Pharmaceutical) containing 4% sucrose is added to a pre-weighed test tube. After the heat sterilization treatment, 0.15 ml of the sample solution and 0.5 ml of the culture solution of Streptococcus mutans 6715 are added, and the culture is performed at 37 ° C. for 20 hours. After completion of the culture, the supernatant is gently removed, and the plaque-like deposits on the test tube wall are washed three times with distilled water 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 measured weight w and W of the plaque-like deposit, a plaque formation inhibition rate is calculated by the following formula.
[0037]
Plaque formation inhibition rate (%) = (1−w / W) × 100
The above measurement is performed while changing the concentration of the sample solution stepwise, and the concentration IC _{50 of the} sample solution at which the inhibition rate is 50% is obtained by interpolation.
The results are shown in Table 2.
[0038]
[Table 2]
Inhibition of plaque formation

[0039]
Example 3
The following mixture was tableted with a tableting machine to produce a health food for caries prevention.
N-vanillyl palmitic acid amide 0.1 parts by weight olive oil 5 parts by weight raffinose 172 parts by weight dextrin 15 parts by weight glycerin fatty acid ester 8 parts by weight
Example 4
The following raw materials were mixed to produce a toothpaste with a dental caries-preventing property.
N-vanillyl stearamide 0.1 parts by weight olive oil 1 part by weight dicalcium phosphate 45 parts by weight CMC sodium salt 1 part by weight glycerin 20 parts by weight sodium lauryl sulfate 2 parts by weight 1-menthol 1 part by weight water 30 parts by weight 0041
Example 5
The following raw materials were processed by a conventional method for producing chewing gum to produce a chewing gum having a caries-preventing action.
N-vanillyl oleic acid amide 0.1 part by weight chewing gum base 20 parts by weight sucrose 50 parts by weight starch syrup 20 parts by weight softener 4 parts by weight perfume 1 part by weight
Example 6
The following raw materials were mixed, concentrated and molded in accordance with a conventional method for producing cocoons to produce a candy having a caries-preventing action.
Spicy capsaicin analog mixture * 1 part by weight sucrose 70 parts by weight waterpox 30 parts by weight citric acid 1 part by weight fragrance 0.1 parts by weight water 15 parts by weight (* obtained by reacting capsaicin and olive oil in the presence of lipase In the unreacted olive oil, 41% by weight of N-vanyl oleic acid amide and 7% by weight of N-vanillyl linoleic acid amide are contained.)
[0043]
Example 7
1 kg of flour, 100 g of corn starch, 250 g of sugar, 125 g of margarine, 5 g of sodium chloride, 25 g of sodium carbonate, 9 g of ammonium carbonate, 6 g of lecithin, 75 g of whole egg, 50 g of calcium lactate, 2 g of spicy capsaicin analog mixture, and 350 g of water Thereafter, it was spread, molded and roasted by a conventional method to produce a biscuit with anti-cavity measures. The blended spicy capsaicin analog mixture was obtained by reacting capsaicin and corn oil in the presence of lipase. N-vanillyl oleic acid amide 18% by weight, N-vanillyl in unreacted olive oil Contains 39% by weight of linoleic acid amide.
[0044]
【The invention's effect】
According to the present invention using a non-savory capsaicin analog, an excellent caries-preventing effect can be achieved by the non-savory capsaicin analog having both a glucosyltransferase inhibitory action and a plaque formation inhibitory action.
[0045]
In addition, the food and drink provided with a caries preventive action according to the present invention has useful effects such as an appetite enhancing action, vasodilation / contraction action, energy metabolism enhancing action and the like of the spicy capsaicin analog, which is extremely useful for health promotion. It will be meaningful.

Claims (3)

Effective at least one selected from N-vanillyl oleic acid amide, N-vanillyl linoleic acid amide, N-vanillyl myristic acid amide, N-vanillyl palmitic acid amide, and N-vanillyl stearic acid amide A glucosyltransferase inhibitor characterized by containing as a component. Effective at least one selected from N-vanillyl oleic acid amide, N-vanillyl linoleic acid amide, N-vanillyl myristic acid amide, N-vanillyl palmitic acid amide, and N-vanillyl stearic acid amide Plaque formation inhibitor characterized by containing as a component.   An oral preparation comprising the glucosyltransferase inhibitor according to claim 1 or (and) the plaque formation inhibitor according to claim 2.