JP2009179631A - Composition having anti-dental caries function - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a material having an anti-dental caries function, particularly a food and drink composition and an oral composition which reduce the cause of dental caries by dental recalcification or the like. <P>SOLUTION: The food and drink composition and the oral composition have an anti-dental caries function and comprise a buffering agent, and the buffering agent is neither (1) a phosphorylated oligosaccharide which is prepared from potato starch and is a glucan composed of α-1,4-bonded 3-5 glucoses to which one phosphoric acid group is bonded nor (2) a phosphorylated oligosaccharide which is prepared from potato starch and is a glucan composed of α-1,4-bonded 2-8 glucoses to which two phosphoric acid group are bonded. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

The present invention relates to a composition for eating and drinking and an oral composition having an anti-cariogenic function. More specifically, the present invention relates to a composition for eating and drinking and an oral composition having an anti-cariogenic function that reduces the occurrence of dental caries due to remineralization of teeth.

Caries means that organic acids produced by oral bacteria present on the tooth surface are prevented from spreading due to some obstruction and the teeth are exposed to high concentrations of organic acid, thereby demineralizing the tooth surface. Will occur. In this sense, all oral bacteria having the ability to ferment sugars that produce organic acids by metabolism can be the cause. A substrate convenient for organic acid production is a saccharide, which includes monosaccharides such as glucose and sucrose, oligosaccharides, and polysaccharides such as starch, which is a polymer of monosaccharides.

Factors that prevent the diffusion of organic acids can be broadly divided into: (1) retention of starch ingested by the meal in the neck and roots, and (2) sugars that are easily degraded such as sucrose (ie, fermentable The insoluble glucan produced by bacteria using sugar as a substrate is fixed to the tooth surface.

The above factor (1) is considered to be caused by all bacteria having sugar-fermenting ability present in the oral cavity such as lactobacilli. The progress of caries in this case is known to be generally slow, and the creation of a high concentration organic acid production environment is highly passive for bacteria.

The above factor (2) is a major factor in modern caries with many sucrose-containing foods.
Streptococcus mutans and Streptococcus sobrinus are considered as the causative bacteria. Both bacteria are a kind of streptococci having a form in which spherical individual bacteria having a diameter of about 0.6 μm are linked in a bead shape. Both bacteria actively produce water-insoluble α-glucan in the presence of sucrose. This glucan has an extremely strong property of adhering to the tooth surface. Moreover, sucrose is rapidly metabolized and exhibits acid-producing ability. In addition, since the bacteria themselves have strong acid resistance, they can survive even under acidic conditions where other bacteria cannot grow. The water-insoluble glucan has the property of firmly binding bacteria to the tooth surface due to its stickiness, and the water-insoluble glucan adsorbed on the tooth surface prevents the organic acid produced by the bacteria from diffusing. An environment is created where the tooth surface is exposed to high concentrations of organic acids. Compared to the case of factor (1), the creation of a high-concentration organic acid production environment is considered to be an active factor for bacteria. The progress of caries in this case is faster than the factor (1).

In consideration of dental health, a new approach to dental caries prevention has been practiced from a microscopic level of tooth decalcification and remineralization (Non-patent Document 1). The tooth surface is composed of calcium and phosphate crystal structure hydroxyapatite [Ca ₁₀ (PO ₄ ) ₆ (OH) ₂ ] and is called enamel. The enamel is the hardest part of the tooth. The organic acid produced by the bacteria in the plaque and the acid contained in the food cause the precious calcium and phosphate to dissolve (demineralize) from the inside of the enamel. Defending. The organic acid penetrates into the enamel through the inter-pillar space filled with moisture and dissolves hydroxyapatite by a process called decalcification. This loss of calcium and phosphate from the enamel tissue results in initial caries beneath the enamel surface. As will be described later, according to the present invention, the caries at this stage can be repaired, and calcium and phosphate ions have penetrated into the subsurface caries and have been lost by a process called remineralization. Apatite can be restored.

The pH of plaque (plaque) tends to become acidic and exceeds the critical pH at which decalcification begins, every time a food or drink containing fermentable carbohydrates is ingested. This is due to the action of acid producing bacteria in the plaque. The pH of the plaque returns to neutral when subjected to the buffering action of saliva, and calcium ions and phosphate ions in the saliva are remineralized into the tooth through the plaque again.

Therefore, as a means of preventing and treating dental caries, it should not be a nutrient source for oral bacteria that cause caries and should not produce organic acids; it should be a nutrient source for mutans bacteria that cause caries. Do not generate water-insoluble glucan and organic acid; prevent the pH from being reduced by this organic acid so as not to exceed the pH at which demineralization begins (for example, it has a buffering action and has a pH
Attention is focused on promoting remineralization.

  Various substances have been known as anti-cariogenic substances.

Caries start when mutans bacteria use sugar as a nutrient source to produce water-insoluble glucan by the action of the enzyme glucosyltransferase. As this glucan covers the tooth surface, plaque is formed. When mutans bacteria undergo acid fermentation inside the plaque, the teeth melt and become decayed teeth.

As non-cariogenic sugars, several oligosaccharides (Non-patent Document 2) that have not been a nutrient source for mutans have already been proposed. An example of such a non-cariogenic sugar is palatinit (Patent Document 1). The remineralization of teeth was promoted by combining palatinit with fluorine or zinc (Patent Document 2). However, paratinite is not preferred for use in foods because of its poor sweetness quality. Also, for the remineralization effect of palatinit,
A high concentration of about 1 to 20% by weight is required.

Sugar alcohols (especially xylitol) are also known as anti-cariogenic substances (for example, Patent Documents 3 and 4). Patent Document 5 discloses an oral composition containing one or more sugar alcohols selected from xylitol, mannitol, galactitol, and inositol. Patent Document 5 also describes that these sugar alcohols can promote not only bacterial growth suppression but also tooth remineralization. Although sugar alcohol is effective only at high concentrations, it is known to induce soft stool by large intake. Further, as described in this example, the effect of xylitol was hardly confirmed.

Furthermore, polyphenol, which is a tea component, has been reported and used as an anti-cariogenic agent (Non-patent Document 3). However, the use of polyphenols also has taste problems and uses are limited.

At present, it is said that fluorine is the most effective substance that exhibits a remineralization effect. Fluorine exhibits sufficient efficacy at about 2 ppm. The following two points have been clarified as the effect of fluorine. Rather, it is used in anticipation of the effect of (2): (1) promotes remineralization; and (2) a strong crystal structure that is difficult to decalcify by incorporating fluorine into hydroxyapatite crystals. Change. In recent years, such fluorine has been added to various oral compositions. For example, Patent Document 6 discloses an oral composition containing calcium carbonate and a soluble fluorine compound. Fluorine ions are known to increase the remineralization ability of fluorine when combined with a sugar alcohol (for example, Patent Documents 7-9). Patent Document 10 discloses a composition containing a mutanase and a fluorine compound as a composition for oral cavity that effectively prevents dental caries by promoting strengthening and remineralization of the tooth.

Promoting dental remineralization by supplying calcium phosphate is also known in the art (eg, Patent Documents 11-12).

Patent Document 13 discloses an oral composition containing calcium carbonate and an alginate. This composition increases the retention of calcium carbonate on the teeth, has a good pH neutralization effect and a remineralization promoting effect, and provides an excellent caries prevention effect.

In Patent Document 14, the phosphorylated oligosaccharide disclosed therein suppresses the calcification phenomenon in which calcium and phosphorus are deposited and crystallizes, and does not become a nutrient source for mutans bacteria that cause caries, and is insoluble in water. The phosphorylated oligosaccharide has a buffering action,
It describes that it also has an effect of preventing a decrease in pH. The above properties prevent tartar development and do not cause plaque and do not cause acid fermentation of mutans bacteria. Phosphorylated oligosaccharides
It is also disclosed that the composition for eating and drinking or the oral cavity composition has an effect of preventing pH from being lowered by lactic acid, which is a fermentation product in plaque, without affecting the flavor. However, Patent Document 14 does not teach that the phosphorylated oligosaccharide can have a remineralization effect at a low concentration as shown in the present specification.

JP 2000-281550 A Japanese Patent Laid-Open No. 2000-247852 JP 2000-128752 A JP 2000-53549 A Japanese Patent Application Laid-Open No. 11-12143 JP-A-11-130463 Japanese Patent Laid-Open No. 11-21217 JP 2000-72638 A JP 2000-154127 A JP-A-8-12541 JP 11-228369 A JP-A-10-310513 JP 11-29454 A JP-A-8-104696

Yoichi Iijima, Takashi Kumagai; Caries Control Mechanism of demineralization and remineralization. Ishiyaku Shuppan Co., Ltd., 21-51, 1999. S. Hamada et al. Jpn. Soc. Starch Sci. 31, 83-91, 1984 S. Sakanaka et al., Fragrance Journal, 11, 42-49, 1990

Therefore, the present invention relates to a material having an anticaries function, in particular, by remineralization of teeth, etc.
It aims at providing the composition for eating and drinking and the composition for oral cavity which reduce generation | occurrence | production of a caries.

The present inventors diligently studied techniques related to caries prevention using various substances. As a result, the present inventors have found a buffer having a tooth remineralization effect and completed the present invention.

In one aspect of the present invention, a composition for eating and drinking having an anti-cariogenic function is provided. Here, the composition includes a buffer having a pH buffering action in the oral cavity, but the buffer is not the following (1) or (2):
(1) A glucan prepared from potato starch, consisting of 3 to 5 glucose linked with α-1,4 and having 1 phosphate group bound thereto, phosphorylated oligosaccharide; or (2) A phosphorylated oligosaccharide, which is a glucan prepared from potato starch, consisting of 2-8 glucose with α-1,4 linkages, and 2 phosphate groups attached.

In one embodiment, the buffer is a phosphorylated oligosaccharide or a sugar alcohol thereof.

In one embodiment, the buffering agent is a phosphorylated oligosaccharide or a sugar alcohol thereof, the phosphorylated oligosaccharide consists of 3-5 glucose α-1,4 linked, and one A glucan having a phosphate group bound thereto, a phosphorylated oligosaccharide, or a glucan having 2 to 8 glucoses linked by α-1,4 and having two phosphate groups bound thereto, provided that Phosphorylated oligosaccharides or sugar alcohols thereof prepared from starch excluding potato starch; chondroitin sulfate; chondroitin sulfate oligosaccharide; glucose-6
-Selected from the group consisting of phosphoric acid; oligogalacturonic acid; and tartaric acid.

In another embodiment, the buffering agent is in the form of an alkali metal, alkaline earth metal, or iron salt. Preferably, the buffer is in the form of a sodium salt or a calcium salt.

In yet another embodiment, the edible composition of the present invention further comprises an anti-caries effective amount of fluorine or a fluorine-containing material.

In another aspect of the present invention, a composition for eating and drinking having an anti-cariogenic function is provided. Here, this composition contains a buffer having a pH buffering action in the oral cavity, a phosphocalcium compensator, a phosphorus preparation and / or a calcium preparation, provided that the buffer comprises the following (1) or (2):
Not a composition:
(1) A glucan prepared from potato starch, consisting of 3 to 5 glucose linked with α-1,4 and having 1 phosphate group bound thereto, phosphorylated oligosaccharide; or (2) A phosphorylated oligosaccharide, which is a glucan prepared from potato starch, consisting of 2-8 glucose with α-1,4 linkages, and 2 phosphate groups attached.

In one embodiment, the buffer is a phosphorylated oligosaccharide or a sugar alcohol thereof.

In one embodiment, the buffering agent is a phosphorylated oligosaccharide or a sugar alcohol thereof, the phosphorylated oligosaccharide consists of 3-5 glucose α-1,4 linked, and one A glucan having a phosphate group bound thereto, a phosphorylated oligosaccharide, or a glucan having 2 to 8 glucoses linked by α-1,4 and having two phosphate groups bound thereto, provided that Phosphorylated oligosaccharides or sugar alcohols thereof prepared from starch excluding potato starch; chondroitin sulfate; chondroitin sulfate oligosaccharide; glucose-6
-Selected from the group consisting of phosphoric acid; oligogalacturonic acid; and tartaric acid.

In another embodiment, the buffering agent is in the form of an alkali metal, alkaline earth metal, or iron salt. Preferably, the buffer is in the form of a sodium salt or a calcium salt.

In yet another embodiment, the edible composition of the present invention further comprises an anti-caries effective amount of fluorine or a fluorine-containing material.

In still another aspect of the present invention, an oral composition having an anti-cariogenic function is provided. Here, this oral composition contains a buffer having a pH buffering action in the oral cavity, provided that the buffer is
A composition that is not (1) or (2) below:
(1) A glucan prepared from potato starch, consisting of 3 to 5 glucose linked with α-1,4 and having 1 phosphate group bound thereto, phosphorylated oligosaccharide; or (2) A phosphorylated oligosaccharide, which is a glucan prepared from potato starch, consisting of 2-8 glucose with α-1,4 linkages, and 2 phosphate groups attached.

In one embodiment, the buffer is a phosphorylated oligosaccharide or a sugar alcohol thereof.

In one embodiment, the buffering agent is a phosphorylated oligosaccharide or a sugar alcohol thereof, the phosphorylated oligosaccharide consists of 3-5 glucose α-1,4 linked, and one A glucan having a phosphate group bound thereto, a phosphorylated oligosaccharide, or a glucan having 2 to 8 glucoses linked by α-1,4 and having two phosphate groups bound thereto, provided that Phosphorylated oligosaccharides or sugar alcohols thereof prepared from starch excluding potato starch; chondroitin sulfate; chondroitin sulfate oligosaccharide; glucose-6
-Selected from the group consisting of phosphoric acid; oligogalacturonic acid; and tartaric acid.

In another embodiment, the buffer is in the form of an alkali metal, alkaline earth metal, zinc or iron salt. Preferably, the buffering agent is in the form of a sodium salt, calcium salt or zinc salt.

In yet another embodiment, the oral composition of the present invention further comprises an anti-cariogenic effective amount of fluorine or fluorine-containing material.

In another aspect of the present invention, an oral composition having an anti-cariogenic function is provided. Here, this composition contains a buffer having a pH buffering action in the oral cavity, a phosphocalcium compensator, a phosphorus preparation and / or a calcium preparation, provided that the buffer comprises the following (1) or (2): Not a composition:
(1) A glucan prepared from potato starch, consisting of 3 to 5 glucose linked with α-1,4 and having 1 phosphate group bound thereto, phosphorylated oligosaccharide; or (2) A phosphorylated oligosaccharide, which is a glucan prepared from potato starch, consisting of 2-8 glucose with α-1,4 linkages, and 2 phosphate groups attached.

In one embodiment, the buffer is a phosphorylated oligosaccharide or a sugar alcohol thereof.

In one embodiment, the buffering agent is a phosphorylated oligosaccharide or a sugar alcohol thereof, the phosphorylated oligosaccharide consists of 3-5 glucose α-1,4 linked, and one A glucan having a phosphate group bound thereto, a phosphorylated oligosaccharide, or a glucan having 2 to 8 glucoses linked by α-1,4 and having two phosphate groups bound thereto, provided that Phosphorylated oligosaccharides or sugar alcohols thereof prepared from starch excluding potato starch; chondroitin sulfate; chondroitin sulfate oligosaccharide; glucose-6
-Selected from the group consisting of phosphoric acid; oligogalacturonic acid; and tartaric acid.

In another embodiment, the buffer is in the form of an alkali metal, alkaline earth metal, zinc or iron salt. Preferably, the buffering agent is in the form of a sodium salt, calcium salt or zinc salt.

In yet another embodiment, the oral composition of the present invention further comprises an anti-cariogenic effective amount of fluorine or fluorine-containing material.

In still another aspect of the present invention, a method for examining a remineralization effect on a tooth of a sample expected to have an anticaries effect is provided. This method comprises a step (A) of causing a calcium precipitation reaction in the presence of this sample from a solution containing phosphorus, calcium, and a tooth component; a step of measuring the calcium concentration or the amount of calcium precipitation in the solution after the precipitation reaction (B Step of causing calcium precipitation reaction from this solution in the absence of this sample (C); Step of measuring calcium concentration in solution or amount of generated calcium precipitate after precipitation reaction; Step (B) and ( A step (E) of measuring a calcium concentration or a precipitation amount in the solution in D)
Is included.

In one embodiment, in the method, the solution comprises hydroxyapatite, buffer, KH ₂ PO ₄ and CaCl ₂ .

As described above, according to the present invention, a composition for eating and drinking and a composition for oral cavity that reduce the occurrence of dental caries due to remineralization of teeth and the like have been provided.

It is a graph which shows the mineral loss amount in the caries in the remineralization test system using a bovine tooth piece. It is a graph which shows the demineralization depth in the remineralization test system using a bovine tooth piece. It is a graph which shows the result of the calcium salt of the phosphorylated oligosaccharide about the remineralization in the simple test system of Example 4. It is a graph which shows the result of the sodium salt of phosphorylated oligosaccharide about the remineralization in the simple test system of Example 4. It is a graph which shows the influence on remineralization in P / Ca = 0.6 using a phosphorylated oligosaccharide. It is a graph which shows the influence on remineralization by the P / Ca density | concentration change in absence of a phosphorylated oligosaccharide. It is a graph which shows the influence on remineralization by the P / Ca density | concentration change in presence of phosphorylated oligosaccharide sodium salt. It is a graph which shows the influence on remineralization by the P / Ca density | concentration change in presence of phosphorylated oligosaccharide calcium salt. It is a graph which shows the remineralization effect of the phosphorylated oligosaccharide calcium salt and sodium salt in Example 5. It is a graph which shows the remineralization effect of xylitol and xylose in Example 5. 6 is a graph showing the remineralization effect of palatinit and palatinose in Example 5. 6 is a photograph showing the results of TLC analysis in Example 7. It is a graph which shows the synergistic effect in the remineralization of the phosphorylated oligosaccharide and fluorine in Example 7. 6 is a photograph showing the results of TLC analysis of phosphorylated oligosaccharides having a standard solution concentration in Example 8. It is a photograph which shows the result of the TLC analysis which shows the elution amount with time at the time of eating the phosphorylated oligosaccharide containing gum in Example 8. It is a graph which shows the remineralization effect of the various substances in Example 12. It is a graph which shows the remineralization effect of the various substances in Example 13. It is a graph which shows the change of pH in the artificial oral cavity apparatus in Example 14. FIG. It is a graph which shows the amount of saliva by POs Ca containing gum in Example 16, or POs Ca non-containing gum chewing. It is a graph which shows the saliva pH value by POs Ca containing gum in Example 16, or POs Ca non-containing gum chewing. It is a graph which shows P content in the saliva by POs Ca containing gum in Example 16, or POs Ca non-containing gum chewing. It is a graph which shows Ca content in the saliva by POs Ca containing gum in Example 16, or POs Ca non-containing gum chewing. It is a graph which shows the change of Ca / P ratio by POs Ca containing gum in Example 16, or POs Ca non-containing gum chewing. It is a graph which shows the demineralization depth (A) and the mineral loss amount (B) in each treatment tooth in Example 16. It is a graph which shows the remineralization rate in Example 17. It is a graph which shows pH of the saliva secreted by ingestion of the POs Ca containing candy in Example 18. It is a graph which shows the quantity of the saliva secreted by ingestion of the POs Ca containing candy in Example 18. It is a graph which shows the quantity of the components Ca and P in the saliva secreted by ingestion of the POs Ca containing candy in Example 18. It is a graph which shows the result of the remineralization test by the POs Ca containing candy in Example 19, and POs Ca soft candy. It is a figure which shows the chemical formula of typical phosphorylated oligosaccharide.

  The present invention is described in detail below.

In the present specification, the anti-cariogenic function includes both a caries prevention function and a caries treatment function. The caries treatment function refers to a function of repairing a part of a tooth once lost by caries. In this specification, having an “anti-cariogenic function” means having one or more of the following properties: (1) having a pH buffering action and reducing pH by an acid produced by oral bacteria. (2) has the ability to suppress the formation of insoluble glucans produced by oral bacteria; (3)
) Ability to promote remineralization of early carious teeth. Preferably it has two of the above properties, most preferably all of the above properties.

According to the composition of the present invention, phosphoric acid and calcium can be stably provided to a carious tooth. Teeth provided with phosphate and calcium are remineralized,
The part of the tooth lost by caries can be restored.

In particular, according to the present invention, since a buffering agent is added to the oral cavity, phosphate and calcium present in the saliva and the like in the oral cavity are stably used for tooth remineralization. Accordingly, it is possible to repair a tooth that was conventionally considered difficult or impossible.

Demineralized lesions, which are the initial symptoms of caries, are restored to a healthy state by replenishing calcium and phosphate (remineralization) in the demineralized enamel part when conditions in the oral cavity are in place. In order to maintain the healthy state of the teeth, it is necessary that minerals are supplied to the affected area of the demineralized lesion by the action of saliva and that demineralization and remineralization at the micro level are balanced. In general, p
When H decreases, the equilibrium relationship of “demineralization-remineralization” breaks down, and the lesion progresses when “demineralization> remineralization”. On the other hand, in the relationship of “remineralization> demineralization”, the demineralized lesion is restored and the teeth are remineralized. The balance between demineralization and remineralization plays an extremely important role in the oral environment, especially saliva and plaque, in the pH, calcium, and phosphate concentrations (Yoichi Iijima, Takashi Kumagai; Mechanism of ash and remineralization.
Ishiyaku Shuppan Co., Ltd., 21-51, 1999. ). According to the present invention, the oral environment can be adjusted to an environment in which remineralization is likely to occur, so that dental caries can be prevented and decalcification lesions, which are the initial symptoms of dental caries, can be treated, and the teeth can be healthy and durable. can do.
In the present specification, the buffering agent refers to a substance that exhibits a pH buffering action in the oral cavity. Specifically, the buffer is a water-soluble salt obtained from, for example, an anion and a cation of the buffer. The buffering agent can stabilize the pH in the oral cavity by being present in the oral cavity. The buffer stabilizes phosphate ions and calcium ions in saliva. For this reason,
In particular, a substance having a good pH buffering action in the presence of phosphate ions and calcium ions is preferable. A buffer that does not inhibit the stability of phosphate ions and calcium ions in an aqueous solution when added to an aqueous solution containing phosphate ions and calcium ions is more preferable. That is, a buffer that easily reacts with phosphate ions or calcium ions to form a precipitate is not preferable.

Furthermore, in the present invention, it is preferable that a pH buffering effect is obtained in the plaque. If pH buffering action is shown in saliva, usually pH buffering action is also shown in plaque. Therefore, a buffer that exhibits pH buffering action in saliva can be used to act pH buffering in plaques. A hydrogen ion sensitive field effect transistor electrode (PH-6010: manufactured by Nihon Koden Kogyo) was installed on an enamel piece, Yoshizumi Tamazawa et al. (Journal of Japan Prosthodontic Society, 40 Special Issue, P147, 1996), Kazuhiko Abe (Dental) Prospect, 90 (3), P650-6
54, 1997), Takahashi-Abbe, S et al. (Oral Micror).
obiol. Immunol. 16, P94-99, 2001), and can be incorporated into the interproximal portion of the lower partial denture to measure the pH in plaque (plaque) formed in the sensitive part. The pH in the plaque is preferably 6 or more, and more preferably 7 or more. When the pH of the plaque is buffered and returns to neutrality, phosphate ions and calcium ions present in the saliva in the oral cavity can be supplied to the tooth surface and remineralized into the tooth. Plaque p
Although the upper limit of H is not particularly limited, it is difficult to consider a high alkali condition in a realistic living body, preferably 10 or less, more preferably 8 or less.

The buffer is usually used in the form of a salt, but may be used in the form of a free acid if necessary. Even if it is provided in the oral cavity in the form of a free acid, there are alkali metals that can form salts with the free acid in the oral cavity, so even if the free acid is provided in the oral cavity, This is because the acid salt is not substantially different from that provided.

Preferred buffering agents that can be used in the present invention can be easily selected by simple experiments. That is, various known pH buffering agents may be added to a neutral aqueous solution (for example, an aqueous solution having a pH of 6 to 8) containing phosphate ions and calcium ions to confirm the presence or absence of precipitation. A pH buffer that does not form a precipitate by such an experiment can be favorably used as a buffer added to the anti-cariogenic composition of the present invention.

In the absence of a buffer, the oral cavity may be acidified due to the influence of organic acids produced by oral bacteria, and saliva or plaque may be acidified. When saliva or plaque is acidified, teeth Ca and P are eluted as Ca ions and P ions, which is one cause of progression of caries. However, if a buffer is present, the pH of saliva and plaque in the mouth
Is stabilized near neutrality, and caries is less likely to progress.

Since saliva usually shows a pH near neutral, a buffer having a good buffering action at a pH near neutral is preferable.

  Preferably, the buffering agent is a substance that does not react with phosphoric acid in saliva to form a precipitate.

  Preferably, the buffering agent is a substance that does not react with calcium in saliva to form a precipitate.

  Preferably, the buffering agent has an acidic functional group.

Preferably, the buffer has either a phosphate group, a carboxyl group, or a sulfate group.

Preferably, the buffer has 3 or less acidic functional groups in the molecule. More preferably, 2
Have no more. When there are too many acidic functional groups in a molecule | numerator, the ability to provide phosphorus and calcium with respect to a hydroxyapatite will fall easily. For example, phosphorylated oligosaccharides having 1 to 2 phosphate groups in the molecule are superior in caries treatment function than phytic acid having 6 phosphate groups in the molecule. Therefore, it is preferable to use a buffer other than such a substance as phytic acid.

A buffer having an excellent ability to provide phosphorus and calcium to hydroxyapatite is preferred. The ability of the buffer to provide phosphorus and calcium to hydroxyapatite can be conveniently tested by a remineralized simple test system method as described below.

Such buffering agents include phosphorylated oligosaccharides and their sugar alcohols. In the present specification, the phosphorylated oligosaccharide refers to a phosphorylated oligosaccharide having at least one phosphate group in the molecule. The number of phosphate groups in the molecule is preferably 3 or less, more preferably 2 or less. In the present application, the neutral oligosaccharide refers to an oligosaccharide having no phosphate group bonded thereto. For example, a phosphorylated oligosaccharide is a glucan composed of 3 to 5 glucoses linked with α-1,4, and one having a phosphate group bonded to the glucan, or a phosphorylated oligosaccharide Sugars are composed of 2 to 8 glucoses linked with α-1,4, and those having two phosphate groups bound to the glucan are included. Examples of such a buffer include acidic saccharides and sugar alcohols thereof (eg, oligogalacturonic acid, chondroitin sulfate, chondroitin sulfate oligosaccharide, glucose-6-phosphate), organic acids (eg, tartaric acid, citric acid, Malic acid, lactic acid, fumaric acid, and maleic acid), nucleic acids (eg, phosphate esters of various nucleosides or nucleotides), amino acids, and sugar alcohols of the above phosphorylated oligosaccharides.

These buffering agents can be in the form of salts, for example metal salts, in order to work effectively. Metals used for such metal salts include alkali metals, alkaline earth metals, zinc, iron, chromium and lead. For example, potassium, sodium, calcium, magnesium and the like can be mentioned. As the metal salt of the buffer contained in the composition for eating and drinking of the present invention, calcium salt and sodium salt are preferable. Moreover, as a metal salt of the buffer contained in the composition for oral cavity of this invention, a calcium salt, a sodium salt, and a zinc salt are preferable. Zinc salts are not used for eating and drinking, but are known as having an effect of preventing bad breath and treating periodontal diseases, and thus are preferable as metal salts contained in oral compositions. Further, the buffer may be an ammonium salt or 4
It may be in the form of a secondary amine salt.

Chondroitin sulfate usually contains one sulfate group per disaccharide. In chondroitin sulfate A, the sulfate group is bonded to the 4-position of N-acetyl-D-galactosamine, and in chondroitin sulfate C, it is bonded to the 6-position of N-acetyl-D-galactosamine. Chondroitin sulfate B (referred to today as dermatan sulfate) is composed of a disaccharide repeating structure consisting of N-acetyl-D-galactosamine-4-sulfate and L-iduronic acid. Chondroitin sulfate is a disaccharide containing unsaturated hexuronic acid at the non-reducing end by chondroitinase,
It can be broken down to oligosaccharides. For example, chondroitinase ABC (Proteus v
ulgaris cells), chondroitinase ACI (Flavobacterium)
heparinum cells), or chondroitinase ACII (Arthrob)
choteroitin sulfate can be broken down into unsaturated disaccharides having hexosamine at the reducing end (however, the latter two enzymes are
It does not act on dermatan sulfate). Chondroitin sulfate and unsaturated oligosaccharides (preferably disaccharides, tetrasaccharides) obtainable by degradation with such enzymes have a remineralization effect.

Oligogalacturonic acid is an oligosaccharide obtained by polymerizing galacturonic acid known as a constituent sugar of pectin, preferably 2 or more, more preferably 3 or more, still more preferably 4 or more, preferably 10 or less, more preferably 8 or less, and even more preferably 6 or less sugars are synthesized.

As used herein, “sugar alcohol” refers to a compound in which the reducing end of a sugar is reduced.
For example, a sugar alcohol of a phosphorylated oligosaccharide can be produced, for example, by hydrogenating the reducing end of the phosphorylated oligosaccharide. Any method known to those skilled in the art can be used for hydrogenation. For example, the oligosaccharide is made into a slightly alkaline solution having a pH of about 8 using 1N sodium hydroxide solution, about 30 ml of about 3% sodium borohydride solution is added to about 100 ml of this solution, and then about 40 ° C. for 1 hour. It can be reduced by standing. In order to industrially produce sugar alcohol, a method using a nickel catalyst known to those skilled in the art can be generally used.

As a buffer contained in the composition for eating and drinking and the composition for oral cavity of the present invention, α-1,4
A glucan composed of 3 to 5 glucose bound, and a phosphorylated oligosaccharide in which one phosphate group is bound to the glucan, or 2 to 8 glucose bound to α-1,4 And a phosphorylated oligosaccharide in which two phosphate groups are bound to the glucan.

Such phosphorylated oligosaccharides can be prepared from common crude plant starches, preferably starches with many phosphate groups attached. Starch-origin plants that can be used to produce phosphorylated oligosaccharides include potato, sweet potato, cassava, corn, wheat, uruchigome, mochigome, mochi corn, mochi wheat, mochi potato, kudzu, yam, lily, chestnut Etc. Among these, rhizomes, rice, wheat and the like contain a lot of bound phosphate groups and are suitable as raw materials. For example, in potato starch, a relatively large amount of phosphate groups are ester-bonded at the 3rd and 6th positions of glucose constituting the starch. Phosphate groups are mainly present in amylopectin. As starch that can be used to produce phosphorylated oligosaccharides, modified starch can also be suitably used. The modified starch is a starch obtained by chemically binding phosphorus to the natural starch as described above. For example, phosphorylated oligosaccharides can be prepared by chemically binding phosphorus to starch such as corn and waxy corn.

The phosphorylated oligosaccharide contained in the composition for eating and drinking and the composition for oral cavity of the present invention can be produced as follows.

For enzymatic degradation of starch and the like, α-amylase (EC 3.2) which is a starch degrading enzyme is used.
. 1.1), β-amylase (EC 3.2.1.2), glucoamylase (EC 3.2.
1.3), isoamylase (EC 3.2.1.68), pullulanase (EC 3.2.1.
41), and neopullulanase (Kuriki et al., Journal of Bacte
riology, 170, 1554-1559 (1988), and cyclodextrin glucanotransferase (EC 2.4.1.19; hereinafter abbreviated as CGTase), which is a glycosyltransferase, is allowed to act on one or more of them. Or one or more of them and α
-Use glucosidase (EC 3.2.1.20) together.

Α-1,6 in starch by degrading with isoamylase or pullulanase
By cutting the branched structure, a phosphorylated saccharide having no branched structure can be obtained. If isoamylase or pullulanase is not used, a phosphorylated saccharide having an α-1,6 branched structure can be obtained. it can. In addition, by decomposing with glucoamylase, non-phosphorylated glucose bound to the non-reducing end can be sequentially released. By performing such enzyme treatment, the number of phosphate groups per molecular weight of the phosphorylated saccharide after purification can be increased or decreased.

Enzymatic degradation can proceed simultaneously by reacting multiple types of enzymes simultaneously.
In short, starch as a raw material is dissolved in water or a buffer adjusted to a pH at which an enzyme can act. To this reaction solution, liquefied α-amylase, pullulanase, glucoamylase and the like are simultaneously added and reacted by heating. When this method is used, the neutral sugar is released while gelatinizing the starch, the non-phosphorylated glucose bound to the non-reducing end of the phosphorylated sugar, or the raw material in the phosphorylated sugar structure Α-1,6 branched structure derived from can be cleaved. This method is not a two-step reaction,
A phosphorylated saccharide with an increased phosphate content is obtained in a one-step reaction.

In the case of causing an enzyme reaction of two or more stages by causing plural types of enzymes to act in individual steps, the order of the enzymes to be actuated is not specified. However, when the starch concentration is high, it is preferable to first act on enzymes including liquefied amylase. First, when isoamylase or pullulanase is allowed to act, the amylose content increases. Since amylose is more susceptible to aging and precipitation than amylopectin, the raw material is aged and precipitated. And it is not affected by other enzymes.

The origin of the amylolytic enzyme, glycosyltransferase, and α-glucosidase used is not particularly limited. For example, as the origin of α-amylase, a starch degrading enzyme preparation derived from Bacillus or Aspergillus can be preferably used. The reaction conditions for the enzyme may be any temperature and pH at which the enzyme can act. For example, a temperature of 25 ° C. to 70 ° C. and a pH of 4 to 8 are preferably used.

First, starch as a raw material is dissolved in water or a buffer adjusted to a pH at which an enzyme can act. Liquefied α-amylase is added to this solution and heated to react to liquefy the starch while gelatinizing. Then, hold | maintain for suitable time at the temperature of 20-80 degreeC. The amount of liquefied α-amylase to be acted may be small or excessive as long as starch can be liquefied. A suitable amount is 20 to 50,000 U. The holding time at this time is not limited as long as the starch is liquefied to such an extent that aging does not occur in the subsequent steps. Preferably, it hold | maintains at 20-80 degreeC for 30 minutes.

It is not necessary to inactivate the enzyme after completion of liquefaction, but the enzyme may be inactivated by a conventional method such as holding at 100 ° C. for 10 minutes. Further, insoluble matters may be separated and removed by a conventional method such as centrifugation or membrane filtration. Thereafter, the phosphorylated saccharide may be fractionated, but in order to obtain a phosphorylated saccharide having an increased phosphate content, the following operation is further performed.

Briefly, after liquefying the raw material, glucoamylase, isoamylase, pullulanase, and α-glucosidase are added simultaneously or in an appropriate order to saccharify, for example, at a temperature of 40-60 ° C. for 30 minutes. Allowed to act for ~ 40 hours to liberate non-phosphorylated glucose bound to the non-reducing end of neutral and phosphorylated sugars from the raw material;
Then, the α-1,6 branched structure derived from the raw material in the phosphorylated saccharide structure can be cleaved. When the glucoamylase, isoamylase, and pullulanase are used in combination, the combination and order of addition are not limited. Moreover, the addition amount and retention time of the enzyme can be determined according to the phosphoric acid content required for the phosphorylated oligosaccharide. Preferably, the glucoamylase is 5
0 to 700 U, 2 to 100 U of isoamylase and pullulanase, and 50 to 700 U of α-glucosidase can be added. The enzyme can be suitably used even if it is immobilized.

After the reaction of each enzyme is completed, it is not necessary to deactivate the enzyme.
The enzyme may be inactivated by a conventional method such as holding for 5 minutes. Further, insoluble matters may be separated and removed by a conventional method such as centrifugation or membrane filtration.

In order to purify the phosphorylated oligosaccharide from the sugar mixture containing the phosphorylated oligosaccharide, an anion exchange resin can be used because the phosphorylated oligosaccharide is an ionic substance unlike the neutral sugar. The type of resin is not particularly limited, but Chitopearl BCW 2500 type (manufactured by Fujibo), Amberlite IRA type (manufactured by Organo), DEAE-cellulose (manufactured by Whatman), DEAE-Sephadex, QAE-Sephadex (Pharmacia) Manufactured), QAE-cellulose (manufactured by Bio-Rad) and the like can be suitably used. The resin is equilibrated with a buffer adjusted to an appropriate pH. For example, about 10-50 mM acetate buffer (
Conditions such as pH 4-5) may be suitably used. The equilibrated resin is packed into a column and charged with a sugar mixture containing phosphorylated oligosaccharides. After washing away the neutral sugar, the adsorbed phosphorylated oligosaccharide is eluted using an alkaline solution or a salt solution.

When elution of phosphorylated oligosaccharide is performed by increasing the ionic strength of the eluate, the type of salt used is not particularly limited. For example, salts such as sodium chloride, ammonium bicarbonate, potassium chloride, sodium sulfate, ammonium sulfate can be preferably used.

When elution of phosphorylated oligosaccharide is performed by changing the pH of the eluate to alkali, the type of alkali reagent used is not particularly limited. For example, ammonia, sodium carbonate, or sodium hydroxide can be used. However, under strong alkaline conditions, the phosphate group is released from the sugar or the reducing end of the sugar is oxidized. Therefore, preferably, the phosphorylated oligosaccharide is eluted at a pH in the range of weakly acidic to weakly alkaline, more preferably pH 3 to pH.
Perform in the range of 8.

In this case, the salt concentration or pH of the eluate is gradually increased, or the phosphorylated oligosaccharide is eluted by gradually increasing the salt concentration or pH, thereby binding per molecule of phosphorylated sugar. It becomes possible to fractionate phosphorylated oligosaccharide components according to the number of phosphate groups.

To purify phosphorylated oligosaccharides from a sugar mixture containing phosphorylated oligosaccharides, activated carbon can also be used instead of anion exchange resin. The type of activated carbon used is not particularly limited,
Preferably, granular activated carbon that can be packed in a column is used. Activated carbon is prepared using a buffer solution, an acid, an alkali, a salt solution, and distilled water so that the neutral sugar excluding glucose can be adsorbed. For example, an activated carbon having a uniform particle size and degassed packed in a column and washed with distilled water can be suitably used. Phosphorylated oligosaccharides can be obtained in a flow-through fraction by applying a sample to the column and adsorbing neutral sugars.

To purify phosphorylated oligosaccharides from a sugar mixture containing phosphorylated oligosaccharides, one carbon atom
Methods of adding ~ 3 alcohols to precipitate phosphorylated oligosaccharides can also be used.
Briefly, by adding alcohol to the sample solution, only phosphorylated oligosaccharides can be obtained as a precipitate. If the sugar concentration is 10% or more, it is desirable to add 3 times or more alcohol by volume.

In the presence of a metal salt, preferably a calcium salt or an iron salt, in addition to the alcohol, the phosphorylated oligosaccharide forms a phosphorylated oligosaccharide metal salt, which tends to precipitate. For this reason,
In the presence of a metal salt, the phosphorylated oligosaccharide can be easily recovered with a small amount of alcohol, as compared with the above-described precipitation only with alcohol. It is preferably carried out under alkaline conditions. Although the kind of salt to be used is not particularly limited, for example, calcium chloride, magnesium chloride, or ferrous chloride has good solubility and can be suitably used. The precipitation generated by adding alcohol is collected by a commonly used method such as decantation, filtration, and centrifugation.

From the phosphorylated oligosaccharide metal salt obtained as a precipitate by adding a metal salt, the phosphorylated oligosaccharide can be produced by removing the metal salt. The removal (desalting) of the metal body can be performed by a conventional method. Desalting can be easily performed by using, for example, a desktop desalting apparatus Microacylator G3 (manufactured by Asahi Kasei Corporation).

The phosphorylated oligosaccharide solution, phosphorylated oligosaccharide, or phosphorylated oligosaccharide derivative thus obtained is subjected to a commonly used drying method such as hot air drying, fluidized bed drying, or vacuum drying. It can be concentrated or powdered. By removing alcohol as necessary, a phosphorylated oligosaccharide that can be used for food and drink or oral application can be obtained.

Phosphorylated oligosaccharides prepared from potato starch using various amylases have phosphoric acid groups that have relatively many phosphate groups at the 3rd and 6th positions of glucose in the potato starch. The group is mainly in position 3 and / or 6 of the glucose residue
It can be an oligosaccharide attached to a position. For example, a phosphorylated saccharide obtained by allowing glucoamylase to act on potato starch can be cleaved from the non-reducing end side until immediately before the phosphate group is bonded to the 6-position of glucose. Therefore, this phosphorylated saccharide has a structure in which glucose bonded at the 6-position is linked to the 6-position of an oligosaccharide having at the non-reducing end or at least the second glucose from the non-reducing end. If the phosphate group is bonded to the 3rd position of glucose, the structure is bonded to the 3rd position of the second glucose from the non-reducing terminal side. Representative examples of phosphorylated oligosaccharides obtained by hydrolyzing potato starch using various amylases are shown in FIG. Of course, the phosphorylated oligosaccharide having the above structure is not limited to those produced by hydrolyzing potato starch using various amylases, and those having the same structure are similarly anti-cariogenic. It has a function.

In the present specification, the sugar alcohol of the phosphorylated oligosaccharide refers to a compound in which the reducing end of the phosphorylated oligosaccharide is reduced. The sugar alcohol of the phosphorylated oligosaccharide can be produced, for example, by hydrogenating the reducing end of the phosphorylated oligosaccharide. Any method known to those skilled in the art can be used for hydrogenation. For example, the oligosaccharide is made into a slightly alkaline solution having a pH of 8 using a 1N sodium hydroxide solution, and a 3% sodium borohydride solution 30 is added thereto.
After adding ml, it can be reduced by standing at 40 ° C. for 1 hour. In order to industrially produce sugar alcohol, a method using a nickel catalyst known to those skilled in the art can be generally used.

The phosphorylated oligosaccharide or sugar alcohol can be in the form of a metal salt. Metals used to form such metal salts include alkali metals, alkaline earth metals, zinc, iron, chromium, lead and the like. For example, potassium, sodium, calcium, magnesium and the like can be mentioned. As a metal salt of the phosphorylated oligosaccharide contained in the composition for eating and drinking of the present invention, a calcium salt and a sodium salt are preferable. Moreover, as a metal salt of the phosphorylated oligosaccharide contained in the composition for oral cavity of this invention, calcium salt, sodium salt, and zinc salt are preferable. Zinc salts are not used for eating and drinking, but are known as having an effect of preventing bad breath and treating periodontal diseases, and thus are preferable as metal salts contained in oral compositions. Furthermore, the phosphorylated oligosaccharide or sugar alcohol may be in the form of an ammonium salt or a quaternary amine salt. It may be an ammonium salt or a quaternary amine salt.

Such a metal salt can be produced as follows. The precipitation of phosphorylated oligosaccharide salt, which is a compound of phosphorylated oligosaccharide and metal salt, can be obtained by alcohol precipitation as described above. If necessary, the operation of redissolving the recovered precipitate in water or a suitable solution and adding the alcohol again may be repeated. By this operation, impurities such as neutral sugars and excess salts can be removed. Ultrafiltration membranes can also be used to remove impurities such as salts.

The above phosphorylated oligosaccharides are known to have the following properties: (1) Not assimilated by cariogenic bacteria (eg, S. mutans, S. sobrinus); (2)
The pH reduction due to sucrose utilization of these bacteria is suppressed in a concentration-dependent manner; and (3) This suppression is due to the buffering action of phosphorylated oligosaccharides (see JP-A-8-104696). It has also been found according to the invention that salt-form phosphorylated oligosaccharides and their sugar alcohols have the effect of promoting tooth remineralization at very low concentrations. By using such properties of phosphorylated oligosaccharides, a composition for eating and drinking and an oral composition having an anti-cariogenic function different from the conventional ones can be obtained. In particular, it is very preferable that it is sufficiently effective at a low concentration for remineralization in terms of addition to food.

In the composition for eating and drinking and the composition for oral cavity of the present invention, the buffering agent is contained in an amount required for the anti-cariogenic function to work effectively in the oral cavity. For example, in the case of phosphorylated oligosaccharide sodium salt, its concentration that may be present in the oral cavity is 0.01-20%, preferably 0.03.
The added amount may be ˜1%. For example, in the case of phosphorylated oligosaccharide calcium salt,
The added amount may be 0.01 to 20%, preferably 0.03 to 1%, in the oral cavity. For example, in the case of a phosphorylated oligosaccharide zinc salt, it may be added such that its concentration that can be present in the oral cavity is 0.01 to 20%, preferably 0.03 to 1%. Phosphorylated oligosaccharide sodium salt, phosphorylated oligosaccharide calcium salt, and phosphorylated oligosaccharide zinc salt are present in the oral cavity when inorganic calcium and phosphorus are in the oral cavity at about 1.5 mM and 0.9 mM, respectively. It may be most preferred that the concentration possible is about 0.2%.

These addition amounts can be determined in consideration of the residence time in the oral cavity of the food-drinking composition and oral cavity composition of the present invention. For example, the case of the food-drinks composition which requires a chewing behavior is demonstrated. For example, in the case of a chewing gum containing about 20% phosphorylated oligosaccharide, about 10 after chewing
Due to elution from the content of the food and drink composition, a relatively high concentration (about 1% to about 5%) of the phosphorylated oligosaccharide can be present in the oral cavity. On the other hand, after about 20 to 30 minutes, 0
. Only 25% or less of the phosphorylated oligosaccharide is present in the oral cavity. Therefore, the phosphorylated oligosaccharide concentration in the oral cavity is diluted to a quarter or less of the concentration in food. Therefore, in the case of such food, a buffering agent can be added to the food at a concentration of 4 times or less, for example, 1 to 4 times the target oral concentration. On the other hand, compositions that cannot require chewing behavior (for example, drinking water) have a residence time in the oral cavity of 1 minute or less. Such compositions are hardly diluted in the oral cavity. For this reason, it can mix | blend in a composition with the density | concentration of 0.1%-5.0% of the same level as the target intraoral density | concentration substantially similarly to the target intraoral density | concentration. As long as the above-mentioned amount in the mouth can be maintained, the buffer may be contained alone or in combination in the composition for eating and drinking and the composition for oral cavity of the present invention.

In another aspect, the food and beverage composition and oral cavity composition of the present invention are also combined with one or more of phosphocalcium compensator, phosphorus preparation or calcium preparation, in addition to the above-mentioned buffer. Including. In particular, in the case of compositions comprising the calcium salt of this substance, the calcium salt can release excess calcium and change the ratio of calcium to phosphorus in the composition. Also, this added buffer can affect calcium elution from the teeth. Here, if the phosphorus: calcium concentration ratio in saliva in the oral cavity that changes due to this buffering agent is compensated, tooth remineralization can be made more effective. In the case of a normal human body, the molar ratio of phosphorus: calcium in saliva (hereinafter referred to as “Ca / P”) is generally 0.25 to 0.67 (P / Ca = 1.45-3). .9) and there is an excess of phosphorus (ie, approximately 3 moles of phosphorus to 2 moles of calcium to 3.9 moles of phosphorus to 1 mole of calcium)
. On the other hand, hydroxyapatite (this is Ca ₁₀ (PO ₄ ) ₆ (O
H) represented by ₂ ) is 1.67 (P / Ca = 0.6), and in the composition constituting the tooth enamel, Ca / P is 1.0-1. 67 (P / Ca = 0.6 ~
1.0). Therefore, Ca / P is 1.0 to 1.67 (P / Ca = 0.6 to 1.0),
By supplying phosphorus and / or calcium together with a buffer so as to be preferably close to 1.67 (P / Ca = 0.6), crystallization of these substances into hydroxyapatite can be promoted.

In the present specification, a substance capable of compensating for Ca / P in this way is referred to as a “phosphocalcium compensator”. As such a calcium phosphate compensator, primary calcium phosphate, secondary calcium phosphate, tertiary calcium phosphate, calcium pyrophosphate, hydroxyapatite powder, amorphous calcium phosphate, bovine bone calcium, eggshell calcium,
Examples include salmon calcium, pearl calcium, fish shell calcium, and α-tricalcium phosphate. Here, Ca / P compensation means that Ca / P is substantially 1.0 to 1.67 (described above).
P / Ca = 0.6 to 1.0). In this case, it is not strictly necessary to be 1.0 to 1.67 (P / Ca = 0.6 to 1.0), and substantially 1.0.
1.0-1.67 (P / Ca = 0.6-1.0) as long as the value can be approximated.
P / Ca = 0.6 to 1.0) may be exceeded. The amount of the compensation agent necessary for compensation varies depending on the type of the buffering agent and the compensation agent to be used.
The range can be easily determined. The calcium phosphate compensator is suitably added in an amount of 1/20 to 20 times, preferably 1/2 to 2 times the molar amount of the phosphate phosphate compensator.

Since there is an excess of phosphorus in saliva, Ca / P is adjusted to 1.0-1 by adding a calcium preparation.
. A case of adjusting to a ratio of 67 (P / Ca = 0.6 to 1.0) is also conceivable. Since the amount of phosphorus in human saliva is 3 to 3.5 mM and the amount of calcium is 0.9 to 2 mM, it is preferable to add about 4 to 5 mM of calcium. Therefore, the buffer calcium salt can be used as a phosphocalcium compensator. In the case of phosphorylated oligosaccharides containing 3% calcium, the addition of about 0.7% phosphorylated oligosaccharide calcium is appropriate. The calcium preparation is not particularly limited, but calcium carbonate, calcium chloride, calcium lactate, calcium gluconate, whey calcium, organic acid calcium, colloidal calcium carbonate, casein phosphopeptide calcium, calcium fluoride, etc. are preferably used. The

Phosphorous formulations may also be included in the food and beverage compositions and oral compositions of the present invention. A phosphorus formulation means a phosphate compound. Examples of such phosphoric acid compounds include sodium phosphate, sodium hydrogen phosphate, potassium phosphate, potassium hydrogen phosphate and the like.

The phosphocalcium compensator, phosphorus preparation, or calcium preparation has a Ca / P of 1.0.
˜1.67 (P / Ca = 0.6-1.0), preferably 1.67 (P / Ca = 0.6), alone or in combination so as to be close to 1.67 (P / Ca = 0.6) And can be added to the oral composition.

The composition for eating and drinking of the present invention is a generic term for human food, animal or fish feed, and pet food. In other words, liquid and powdered beverages such as coffee, tea, Japanese tea, oolong tea, juice, processed milk, sports drinks, bakery items such as bread, pizza, pie, baked confectionery items such as cookies, crackers, biscuits, cakes, Pasta such as spaghetti, macaroni, noodles such as udon, soba, ramen, confectionery such as candy, soft candy, gum, chocolate, snacks such as oysters, potato chips, snacks, frozen confectionery such as ice cream, sorbet, Dairy products such as cream, cheese, powdered milk, condensed milk, milk drinks, Western confectionery such as jelly, pudding, mousse, yogurt, buns, uro,
Japanese sweets such as rice cake, rice cake, soy sauce, sauce, noodle soup, sauce, soup stock, stew base, soup base, compound seasoning, curry base, mayonnaise, ketchup and other seasonings, curry, stew , Soups, bowls and other retort or canned foods, ham, hamburger, meatballs, croquettes, dumplings, pilaf, rice balls and other frozen and refrigerated foods, seafood processed foods such as chikuwa and salmon Includes. In addition, baby milk, baby food, baby food, pet food, pet gum, animal feed, sports food, nutritional supplements, which are used for the absorption of nutritionally effective ingredients such as calcium,
Health foods are also included.

In a preferred embodiment, the food or drink is a food or drink that is highly chewed for eating, such as gum. In the case of foods and drinks that are highly chewed, the buffering agent is easily diffused in the oral cavity, and an anti-cariogenic effect is obtained well. In the case of foods and drinks with a high degree of chewing, the buffering agent can be blended in a proportion of preferably 0.1 to 50% by weight, more preferably 0.5 to 20% by weight of the total weight of the food and drinks. %, More preferably 0.5 to 10% by weight,
Especially preferably, it is 0.5 to 5 weight%. Specifically, for example, a gum containing 0.1 to 50% by weight of a buffering agent. Also preferred are tablet confectionery, candy, gummi, etc., containing 0.1 to 50% by weight of a buffering agent.

In another preferred embodiment, the food or drink is a food or drink that requires little chewing to eat, for example, drinking water such as juice or fresh water. In the case of foods and drinks that require little chewing to eat, the buffer is preferably 0.1% of the total weight of the food and drinks.
Can be blended in a proportion of ˜70 wt%, more preferably 0.1 to 50 wt%,
More preferably, it is 0.2 to 5% by weight. Specifically, for example, juice containing 1 to 30% by weight of a buffering agent is used. Also preferred are vegetable juices, natural fruit juices, milk drinks, milk, soy milk, sports drinks, near-water drinks, nutritional drinks, coffee drinks, cocoa and the like in which 0.1 to 10% by weight of a buffering agent is blended.

In yet another preferred embodiment, the food and drink is a food and drink eaten by chewing as much as a normal meal staple food. Food and drink eaten as a staple food are preferred. For example, cooked rice. In the case of foods and drinks to be eaten as staple foods, since they are eaten in large quantities, there is an advantage that the caries prevention effect is large and can be obtained for a long time even if the concentration of the buffering agent added to the food or drink is small. In the case of foods and drinks that are eaten by chewing at the same level as a normal meal staple food, the buffering agent may be blended preferably in a proportion of 0.01 to 20% by weight of the total weight of the foods and drinks. More preferably, it is 0.02 to 10% by weight, further preferably 0.03 to 5% by weight, and particularly preferably 0.05 to 3% by weight. Specifically, for example, rice containing 0.02 to 10% by weight of a buffer, bread containing 0.01 to 20% by weight of a buffer, and the like.

Of course, food and drink other than the food and drink shown in each of the above preferred embodiments may be used. Specifically, for example, ramen containing 0.1 to 20% by weight of a buffering agent, 0.1 to 0.1% of a buffering agent.
Noodles containing 20% by weight, 0.1-20% by weight of buffer and 0% of buffer
. Pretzel containing 1 to 20% by weight, agar containing 0.1 to 20% by weight of buffer, jelly containing 0.1 to 20% by weight of buffer, 0.1 to 20% by weight of buffer Blended yogurt, cookie blended with 0.1-20% by weight buffer, 0.1-20 buffer
Tablets containing 1% by weight, tofu containing 0.1 to 20% by weight of buffer, 0.1 to 10% of buffer
Chocolate containing 20% by weight, rice cracker containing 0.1-20% by weight of buffer, dumplings containing 0.1-20% by weight of buffer, 0.1-20% by weight of buffer Also preferred are hams and the like.

The “oral composition” in the present specification means all compositions other than food and drink that can be introduced into the oral cavity and come into contact with teeth. It may be a pharmaceutical, a quasi-drug, or other. For example, cosmetics are also included in “oral compositions” (
More specifically, toothpastes that have the effects of preventing tooth decay, whitening teeth, removing plaque, purifying the mouth, preventing bad breath, removing the teeth on the teeth, and preventing calculus deposition ( These can be certified as “cosmetics” by the 2001 revised Pharmaceutical Affairs Law)). Specifically, for example, the oral composition of the present invention includes a dentifrice, a mouthwash, a troche, a mouthwash, a gingival massage cream, a throat candy, artificial saliva, and the like.

In one preferred embodiment, the oral composition is a dentifrice, and the buffering agent can be formulated in a proportion of preferably 0.01 to 20% by weight of the total weight, more preferably 0.
. It is 02-10 weight%, More preferably, it is 0.03-5 weight%, Especially preferably, it is 0.05-3 weight%.

In one preferred embodiment, the oral composition is a mouthwash and the buffer is
The total weight is preferably 0.01 to 20% by weight, more preferably 0.02 to 10% by weight, still more preferably 0.03 to 5% by weight, especially Preferably it is 0.05 to 3 weight%.

In one preferred embodiment, the oral composition is an oral ointment, and the buffering agent can be blended in a proportion of preferably 0.01 to 20% by weight of the total weight, more preferably 0. 0.02 to 10% by weight, more preferably 0.03 to 5% by weight, and particularly preferably 0.05 to 3% by weight.

Also preferred are dentifrices, mouthwashes, troches, mouthwashes, artificial saliva, etc., containing 0.1 to 20% by weight of a buffering agent.

Artificial saliva has been used to improve xerostomia and contains components such as minerals so that it is almost identical to human saliva. Sweeteners and preservatives can be added to the artificial saliva. As sweeteners, sugar alcohols and artificial sweeteners are preferably used, and as preservatives, sodium benzoate, sorbic acid and the like are preferably used. Artificial saliva containing the buffer may not only moisten the tongue and oral laryngeal mucosa to smooth the movement of the tongue and mucosa, but also have a preventive and therapeutic effect on caries.

The composition for eating and drinking and the composition for oral cavity of the present invention further contain fluorine as necessary. The food and beverage composition and oral cavity composition of the present invention contain fluorine in an amount not exceeding 1000 ppm, preferably 0.1 to 500 ppm, more preferably 0.1 to 300 ppm. In order to suitably increase the efficacy of fluorine at 100 ppm or more, the buffer is a pharmaceutical,
It is also suitable for use in quasi drugs and cosmetics. The food-drinking composition and oral cavity composition of the present invention can have a higher effect on tooth remineralization by further containing fluorine. As used herein, “fluorine” means fluorine ion. "Fluorine-containing material"
It means any material that provides fluorine ions, preferably a compound containing fluorine ions, such as sodium monofluorophosphate, sodium fluoride, potassium fluoride, ammonium fluoride, fluorinated amine salt, fluoride And tin oxide. The use of sodium monofluorophosphate and sodium fluoride is preferred.

Fluorine or fluorine-containing materials alone are less effective against tooth remineralization, especially 100
Since it is easily insolubilized at a high concentration of ppm or more, its effectiveness is remarkably reduced. However, it has also been revealed by the present invention that the efficacy of fluorine or fluorine-containing materials is increased in combination with a buffer. In addition, in food, tea with a high content of fluorine (200-300pp
m) and the like can be preferably used. Fluorine or fluorine-containing materials can be incorporated into tooth crystals to produce strong crystals that are resistant to acids. For this reason, the composition for eating and drinking of the present invention and the composition for oral cavity can act on the generation of strong crystals of teeth in addition to the remineralization of teeth, thereby reducing the occurrence of caries.

The food and beverage composition and oral composition of the present invention may also contain other substances known to those skilled in the art to have an anti-cariogenic function. Such substances include various oligosaccharides (Panose (6
² -glucosyl-maltose), isomaltoligosaccharide, palatinose (6-O-α-D-
Glucopyranosyl-D-fructofuranose), trehalose (O-α-D-glucopyranosyl (1-1) -α-D-glucopyranoside), maltooligosaccharide, dairy oligosaccharide ^TM (4
^G- β-D-galactosyl sucrose), fructooligosaccharide, coupling sugar, xylosyl fructoside, cyclodextran, etc.); sugar alcohols (xylitol, erythritol, paratinite, sorbitol, maltitol, mannitol, etc.); tea extract (Containing fluorine, polyphenol, catechin, etc.); herbs (eg, mint, peppermint oil, chamomile, sage, ginger, rosemary, etc.) (Shibuy)
a et al., FRAGRANCE JOURNAL SPECIAL ISSUES, 12, p
150-155, 1992); enzymes (eg, dextranase, mutase, etc.);
A vaccine (for example, secretory immunoglobulin A against S. mutans) and the like can be mentioned. Sugar alcohols are preferred, and xylitol is more preferred. The food-drinking composition and oral cavity composition of the present invention can have a further increased caries prevention effect by containing the above substances.

The remineralization effect of the buffer can be examined by the following method. For example, Inaba. D
Et al., Eur. J. et al. Sci. 105: 74-80, 1997, Inaba. D et al. De
nt. Helth. 47: 67-74, 1997, Iijima. Y et al., Caries
Research. 33: 206-213, 1999, remineralization test systems using bovine tooth pieces are known in the art.

In order to investigate the remineralization effect of the buffering agent that can be included in the beverage composition and oral cavity composition of the present invention, the present inventors are simpler than the remineralization test system described above. A test system was developed. Conditions that are likely to cause remineralization include the following: Calcium and phosphorus are quickly supplied to the tooth surface at the contact surface of the tooth surface (hydroxyapatite), and change into a tooth component (hydroxyapatite); Higher concentrations of calcium or phosphorus are maintained in the containing system; and calcium and phosphorus are not precipitated and lost from the system in places other than the tooth surface. By simplifying such conditions that are likely to cause remineralization, in systems containing hydroxyapatite, calcium and phosphorus are supplied for crystallization, and soluble calcium is reduced. In a system that does not contain hydroxyapatite, phosphorus and calcium do not precipitate, and high solubility is maintained. Therefore, the remineralization effect could be examined by comparing the extent of calcium dissolution in these two systems.

This simple test system is described below. As a standard method for quantitatively measuring the distribution of dental mineral concentration, the TMR (Transversal microradiography) method has been used in many studies on demineralization and remineralization. However, there are limitations such as the time required for evaluation and the need for advanced techniques in experimental methods. Therefore, it is desired to develop a simple evaluation system that can quickly grasp changes in the concentration of dental minerals. On the other hand, the remineralization phenomenon in the initial carious lesion of tooth enamel is considered to proceed in the following two processes: Component calcium (Ca) ions and phosphorus (P) ions are supplied to the demineralization section; 2
. The supplied Ca ions and P ions are used for crystal growth of enamel in the demineralized part. In other words, when considering the two processes of remineralization, the remineralization promoting substance is C
It was considered to be a substance that inhibits insolubility and precipitation of aP but does not inhibit HAp crystal growth. This test system has a correlation as described above with a system using bovine teeth as used conventionally, and is a simple and excellent method.

In one aspect of the present invention, the present invention relates to a method for examining a remineralization effect on a tooth of a sample expected to have an anti-cariogenic effect. This method comprises a step (A) of subjecting a solution containing phosphorus, calcium, and a tooth component to a calcium precipitation reaction in the presence of the sample; a step of measuring a calcium concentration or a calcium precipitation amount in the solution after the precipitation reaction (B ); A calcium precipitation reaction from the solution in the absence of the sample (C); after the precipitation reaction, a step of measuring the calcium concentration in the solution or the amount of calcium precipitate formed (D); and steps (B) and ( (E) comparing the calcium concentration or the amount of precipitation in D). In a preferred embodiment, the solution is hydroxyapatite, buffer, KH ₂ PO ₄ and C
aCl ₂ may be included. As the “tooth component” to be included in the solution, any substance capable of depositing phosphorus and calcium to form hydroxyapatite generated by remineralization can be used. Although it is preferred to use hydroxyapatite, mammalian teeth such as cows and sections thereof or crushed pieces thereof may also be used. Regarding the preparation of the solution for the calcium precipitation reaction, the above-mentioned phosphorus, calcium, and tooth components may be added in any order. Preferably, however, the sample is put first and then the phosphorus, calcium, and tooth component suspension or desorption is added. Add in order of ionic water to make a solution. It is preferable to adjust the pH of the solution after adding the KH ₂ PO ₄ solution. The calcium precipitation reaction is usually performed at room temperature for 10 hours to several days (preferably 10 hours to 7 days, more preferably 18 hours to 42 hours).
Generated by incubating. Calcium solubility in solution can be measured using any procedure known to those skilled in the art. The calcium solubility in the solution can be measured, for example, by the OCPC method (using calcium C test Wako manufactured by Wako Pure Chemical Industries, Ltd.). Or the calcium precipitation amount in a solution can also be measured. The amount of calcium precipitation in the solution can be measured using any procedure known to those skilled in the art. Calcium solubility in solution can be measured using any procedure known to those skilled in the art. As such a method, for example, the ICP method (Inductive
(Coupled Plasma method), atomic absorption analysis, ion electrode method and the like.

Further, in order to examine the anti-cariogenic function, an artificial oral cavity device for obtaining a decalcified enamel as close as possible to reality can be used (for an artificial oral cavity device, for example, Jpn.J.Ora).
l Biol. 20: 288-291, 1984). For example, this device
Means may be provided for instilling the electrode, the enamel section mounted around the electrode, and the mutans streptococcal suspension, the media solution, and the carbohydrate solution. By this apparatus, mutans cocci can be fixed on the electrode surface while synthesizing a water-insoluble glucan to form an artificial plaque, and a low pH can be created. Furthermore, similar artificial plaques are formed on the enamel slices, and a clear decrease in the hardness of the enamel is obtained.

The present invention will be specifically described below with reference to examples. This example does not limit the invention. The materials, reagents, etc. used in the examples are available from commercial sources unless specified otherwise.

Example 1
This example shows a method for producing a phosphorylated oligosaccharide for use in the composition of the present invention.

A 1% solution of potato starch was dissolved in a solution containing 5 ml of 6 mM sodium chloride and 2 mM calcium chloride and gelatinized by rapidly raising the temperature to 100 ° C., and then α-amylase (Fuctamirase; manufactured by Hankyu Kyoei Bussan Co., Ltd.) ) For 35 U, 3 at 50 ° C.
Hold for 0 minutes. A small amount of this reaction solution is taken to make a 0.2% sugar solution, and 0.01M iodine-
After adding 1/10 amount of potassium iodide solution and confirming that iodine coloration is negative, pullulanase (manufactured by Hayashibara Biochemical Laboratories) 2U and glucoamylase (manufactured by Toyobo) 6U at the same time 4
It was allowed to act at 0 ° C. for 20 hours. The reaction was stopped, and this solution was subjected to centrifugation, and then the supernatant was applied to an anion exchange resin (Chitopearl BCW2501; manufactured by Fujibo) equilibrated with 20 mM acetate buffer (pH 4.5). Thorough washing with the same buffer was performed to remove neutral sugars, followed by elution with the same buffer containing 0.5 M sodium chloride. Each elution fraction was concentrated using an evaporator, desalted and lyophilized to obtain phosphorylated oligosaccharide.

The phosphorylated oligosaccharide obtained as described above was again applied to an anion exchange resin column (Chitopearl BCW2501) equilibrated with 20 mM acetate buffer (pH 4.5). The column was sufficiently washed with the same buffer to remove neutral sugars. Fractions eluted with the same buffer containing 0.15 M sodium chloride and then with the same buffer containing 0.5 M sodium chloride were collected. As a result of analysis based on the above structure determination method, these fractions were desalted and freeze-dried, so that 3 or more and 5 or less α-1,4 glucose were bound from the 0.15 M sodium chloride elution fraction. Phosphorylated oligosaccharide (PO-1 fraction) in which one phosphate group is bound to glucan is obtained, and from 0.5M sodium chloride elution fraction, 2 or more and 8 or less glucose α-1,4 bonds As a result, phosphorylated oligosaccharide (PO-2 fraction) in which two or more phosphate groups were bonded to the obtained glucan was obtained.

  The structural analysis of the phosphorylated oligosaccharide was performed as follows.

First, the phosphate group was removed from the phosphorylated oligosaccharide. 100 μl of 3% phosphoric acid oligosaccharide solution, 100 μl of 10 mM magnesium chloride, 0.3 mM zinc chloride, and 0.05%
60 mM sodium carbonate buffer (pH 9.4) containing sodium azide was mixed with 100 μl of 30 U / ml alkaline phosphatase (EC 3.1.3.1; E.co.
li origin; Sigma) was added and reacted at 40 ° C. for 18 hours. The reaction is stopped by removing alkaline phosphatase using an ultrafiltration membrane, and a reaction liquid (hereinafter referred to as reaction liquid A) containing a phosphate group-removed sugar (hereinafter referred to as dephosphorylated sugar) as a component. Obtained.

5,000 U / ml β-amylase (derived from sweet potato; manufactured by Sigma) dissolved in 10 μl of 200 mM acetate buffer (pH 4.8) was added to 10 μl of the obtained reaction solution A, and the mixture was incubated at 37 ° C. for 2 hours. (Hereinafter, this solution is referred to as reaction solution B). Similarly, reaction solution A10
300 μg of glucoamylase (derived from Rhizopus; manufactured by Toyobo) dissolved in 10 μl of 60 mM acetate buffer (pH 4.5) was added to μl and maintained at 35 ° C. for 18 hours (hereinafter, this solution was referred to as reaction solution) C).

The reaction liquids A to C were analyzed to confirm the product. These reaction solutions were analyzed by standard maltooligosaccharides of various degrees of polymerization by high performance liquid chromatography using an anion exchange resin column CarboPac PA-100 (φ4 × 250 mm, manufactured by Dionex) or thin layer chromatography using silica gel. It confirmed by comparing with. Elution of dephosphorylated saccharide using high performance liquid chromatography is carried out by increasing the concentration of 1M sodium acetate using 100 mM sodium hydroxide as a basic solution. Detection was performed by pulsed amperometry (Dionex). Analysis of dephosphorylated saccharide by thin layer chromatography was carried out by multiplying dephosphorylated saccharide with acetonitrile / water = 80/20, spraying with a solution of sulfuric acid / methanol = 1/1, and maintaining at 130 ° C. for 3 minutes. It was done by doing.

By analyzing the reaction solution A, the chain length of the phosphorylated oligosaccharide was confirmed. When the reaction solution B was analyzed, only maltose, or maltose and maltotriose (and slightly glucose) were detected. Therefore, it was confirmed that this dephosphorylated sugar is a glucan in which glucose is α-1,4 linked. Further, when the reaction solution C was analyzed, only glucose was detected. Therefore, it was confirmed that this dephosphorylated sugar consists of α-linked glucose.

The average sugar chain length (hereinafter expressed as DP with glucose as one unit) was determined from the sugar content of each degree of polymerization constituting dephosphorylated sugar. The total amount of sugar in the total phosphorylated saccharide is quantified by the phenol sulfate method, and the number of bound phosphate groups is quantified as inorganic phosphoric acid after wet ashing (starch-related carbohydrate experimental method, biochemical experimental method 19, Nakamura Michinori) 31, 1986, Academic Publishing Center). 1
The number of bonded phosphate groups per molecule was calculated from the amount of inorganic phosphate quantified after wet ashing and DP using the following formula.

（実施例２）
リン酸化オリゴ糖の分子内にリン酸基を１個有するＰＯ−１画分およびリン酸基を２個
有するＰＯ−２画分のそれぞれ１０ｇを１００ｍｌの蒸留水に溶解した。これらの水溶液
を電気透析器（旭化成製マイクロアシライザーＧ３、ＡＣ２１０−４００膜）で脱塩し、
強カチオン交換樹脂（日新化成製Ｄｏｗｅｘ５０ｗ、２０−５０ＭＥＳＨ、Ｈ−Ｆｏｒｍ
）でイオン交換した後、ｐＨ２．７の糖溶液を得た。本溶液を１Ｎ水酸化ナトリウム溶液
または水酸化カルシウム溶液を用いて中和した後、凍結乾燥し、リン酸化オリゴ糖のナト
リウム塩またはカルシウム塩を調製した。

(Example 2)
10 g each of the PO-1 fraction having one phosphate group in the molecule of the phosphorylated oligosaccharide and the PO-2 fraction having two phosphate groups were dissolved in 100 ml of distilled water. These aqueous solutions were desalted with an electrodialyzer (Asahi Kasei Microacylator G3, AC210-400 membrane),
Strong cation exchange resin (Nisshin Kasei Dowex50w, 20-50MESH, H-Form
) To obtain a sugar solution having a pH of 2.7. This solution was neutralized with 1N sodium hydroxide solution or calcium hydroxide solution, and then lyophilized to prepare sodium or calcium salt of phosphorylated oligosaccharide.

The phosphorylated saccharide (in the form of sodium salt or calcium salt) used in the following examples is
It is a phosphorylated saccharide mixture containing 80% or more of the PO-1 fraction phosphorylated saccharide, and the remainder being the PO-2 fraction phosphorylated saccharide.

(Example 3)
This example demonstrates verification in a system using bovine teeth to clarify the effect of phosphorylated oligosaccharides on remineralization in early caries.

This experiment is basically based on Inaba. D et al., Eur. J. et al. Sci. 105: 74-80,
1997, Inaba. D et al. Dent. Helth. 47: 67-74, 1997
Iijima. Y et al., Caries Research. 33: 206-213, 19
It carried out based on the description content of 99.

The tooth pieces used in this experiment were prepared as follows: A 3 mm square cow tooth piece was placed with the enamel side up. The surface other than the enamel surface was coated with a composite resin. The enamel was treated with water-resistant paper. The decalcification treatment was performed as follows: The tooth pieces were immersed in a 1% lactic acid gel (pH 5.0) containing 6% carboxymethylcellulose gel at 37 ° C. for 3 weeks. The remineralization treatment was performed as follows: decalcified teeth were treated with 1.5 mM CaCl ₂ ,
20 mM 2- [4- (2-hydroxyethyl)]-1- containing 0.9 mM KH ₂ PO ₄
It was immersed in piperidinylethanesulfonic acid (HEPES) buffer (pH 7.0) at 37 ° C. for 1 week.

The following eight test groups were prepared: (1) Decalcification treatment only (blank; “Bla” in FIGS.
nk "); (2) Remineralization treatment only (negative control;" Control "in FIGS. 1 and 2)
(3) Remineralization solution + 2 ppm fluorine (F) (positive control; “2p” in FIGS.
pmF "); (4) Remineralization solution + 4.0% sodium phosphorylated oligosaccharide ("
POs Na 4% "); (5) Remineralization solution + 1.0% sodium phosphorylated oligosaccharide (
(6) Remineralization solution + 0.2% sodium phosphorylated oligosaccharide (“POs Na 0.2%” in FIGS. 1 and 2); (7) Relime Solution + 0.2%
Phosphorylated oligosaccharide calcium (“POs Ca 0.2%” in FIGS. 1 and 2); and (8)
Remineralized solution + 0.07% phosphorylated oligosaccharide calcium ("POs Ca 0" in Figs. 1 and 2)
. 07% ").

After each treatment, a 200 μm thin piece was prepared from the treated tooth piece, and a mineral concentration distribution analysis was performed from the micrographic image (not shown). When the tooth piece is decalcified, the mineral is eluted and lost, resulting in a cavity (start of caries). A graph of mineral loss by this mineral concentration analysis analysis (Fig. 1, the vertical axis shows mineral loss) and demineralization depth (
FIG. 2 shows the deashing depth (μm) on the vertical axis. According to FIG. 1, both phosphorylated oligosaccharide sodium and phosphorylated oligosaccharide calcium had the least amount of mineral loss at the lowest concentration tested. This amount of mineral loss was less than (2) positive control. Phosphorylated oligosaccharide sodium and phosphorylated oligosaccharide calcium showed a low demineralization depth (FIG. 2). This may indicate that the cavity has been filled by remineralization. Interestingly, the demineralization depth remained deep in the positive control with fluorine addition (2).

After each treatment, an analysis of the calcium and phosphorus concentrations in the post-remineralization solution was also performed. The solution was centrifuged at 10,000 g for 2 minutes and the supernatant was analyzed. The concentration of phosphorus is
By the molybdic acid method (new edition analytical chemistry experiment (first edition), pp. 313 to 314, described in Kagaku Dojin Co., Ltd.), and the calcium concentration was determined by the OCPC method (Wako Pure Chemical Industries, Ltd .:
Measured using “Calcium C Test Wako” kit). The results are shown in Table 1.
Shown in

表１より、リン酸化オリゴ糖の添加によって、溶液中に溶解しているカルシウムおよび
リン濃度が高く維持されていることが分かった。

From Table 1, it was found that the calcium and phosphorus concentrations dissolved in the solution were kept high by the addition of phosphorylated oligosaccharide.

Thus, this experiment shows that the addition of phosphorylated oligosaccharides maintains a high concentration of dissolved phosphorus and calcium in the solution, so that these solubilized phosphorus and calcium are supplied to the carious portion, and It is suggested that it can be used for remineralization. Such a phenomenon is considered to occur in the human oral cavity.

Example 4
This example shows verification in a simple recalcification test system in order to clarify the influence of phosphorylated oligosaccharide on remineralization in the initial caries.

(Remineralization test system procedure)
In order to more easily verify the remineralization phenomenon, the conditions under which remineralization is likely to occur have been simplified. In systems containing hydroxyapatite, calcium and phosphorus are supplied for crystallization, and dissolved calcium is reduced. On the other hand, in a system that does not contain hydroxyapatite, calcium and phosphorus do not precipitate, and their high solubility is maintained. Based on these facts, the following test system was designed.

Add 500 μl volume of solution by adding in the following order: (1) 200 mM
50 μl of HEPES buffer (pH 7.0), (2) 200 μl of deionized water or sample,
(3) 18 mM KH ₂ PO ₄ solution 50 μl, (4) 30 mM calcium chloride solution 50 μl
And (5) Hydroxyapatite suspension (5 mg / ml) or deionized water. (3
) Is added, and the pH of the solution is adjusted with 0.1N potassium hydroxide solution. The resulting solution is stirred and incubated at 37 ° C. for 1-7 days. Next, the mixture is centrifuged at 12,000 rpm for 3 minutes, and the calcium concentration in the obtained supernatant is measured using the OCPC method (same as above). This measures the absorbance at 570 nm using Calcium C Test Wako (Code; 27-21801). The percent of solubilized calcium is obtained by multiplying the calcium concentration in the supernatant divided by the added calcium concentration by 100. The percent remineralization is determined by determining the difference between the value obtained with deionized water in (5) and the value obtained when adding hydroxyapatite.

(Effects of various concentrations of phosphorylated oligosaccharides on remineralization)
Using the simple test system described above, various concentrations of phosphorylated oligosaccharide sodium and calcium salts were incubated at 37 ° C. for 18 or 42 hours. The results of the sodium salt of phosphorylated oligosaccharide and the calcium salt of phosphorylated oligosaccharide for remineralization are shown in FIG. 3 and FIG. 4, respectively (both FIG. 3 and FIG. %
), The horizontal axis indicates the sample (%), the control indicates no sample addition; at each sample concentration, the left bar is treated for 18 hours and the right bar is treated for 42 hours). Phosphorylated oligosaccharide sodium salt had a high ability to dissolve added calcium even at low concentrations (FIG. 3). Phosphorylated oligosaccharide calcium salts have a low ability to further dissolve extraneous additive calcium salts, but rather release excess calcium and change the ratio of calcium / phosphorus in the solution to produce a new precipitate As a result, there is a tendency that the calcium concentration in the system cannot be maintained high (FIG. 4).

Therefore, phosphorylated oligosaccharide sodium salt was able to exhibit a dissolving action without changing the concentration ratio of calcium and phosphorus in the system. In the case of phosphorylated oligosaccharide calcium salt, the ratio of Ca / P is set to 1 by simultaneously supplying phosphorus (phosphoric acid, phosphorus compound, etc.).
. It was considered necessary to maintain 67 (corresponding to P / Ca ratio = 0.6). Alternatively, it is necessary to set the addition concentration so as not to affect the ratio.

(Influence on remineralization effect at Ca / P concentration ratio = 1.67 (corresponding to P / Ca concentration ratio = 0.6) using phosphorylated oligosaccharide)
When the calcium / phosphorus concentration ratio is Ca / P concentration ratio = 1.67 (P / Ca concentration ratio = 0.6) and the calcium salt of PO is used, the calcium source is phosphorylated oligosaccharide. And the concentration of phosphorus was set. A sodium salt was also set according to the phosphorylated oligosaccharide concentration. The concentration settings are shown in Table 2 below.

上記簡易試験系を使用して、３７℃で１５時間インキュベートした。この結果を図５に
示す（縦軸に再石灰化促進率（％）を示し、横軸にＣａ濃度（ｍＭ）を示し、黒四角は、
リン酸化オリゴ糖塩無添加のコントロールを、白菱形は、リン酸化オリゴ糖ナトリウム塩
（ＰＯｓ Ｎａ）を、白丸は、リン酸化オリゴ糖カルシウム塩（ＰＯｓ Ｃａ）を表す）
。図５に示したように、Ｃａ／Ｐ濃度比＝１．６７（Ｐ／Ｃａ濃度比＝０．６）の比率を
一定にして、カルシウムの添加濃度を上昇させた場合には、リン酸化オリゴ糖のナトリウ
ム塩およびカルシウム塩ともに、同様な結果が得られた。添加したカルシウム塩が６ｍＭ
以上になると、リン酸化オリゴ糖の添加による効果は低下した。

Incubation was carried out at 37 ° C. for 15 hours using the above simple test system. The results are shown in FIG. 5 (the vertical axis shows the remineralization promotion rate (%), the horizontal axis shows the Ca concentration (mM),
(Phosphorylated oligosaccharide salt-free control, white rhombus represents phosphorylated oligosaccharide sodium salt (POs Na), white circle represents phosphorylated oligosaccharide calcium salt (POs Ca))
. As shown in FIG. 5, when the ratio of Ca / P concentration ratio = 1.67 (P / Ca concentration ratio = 0.6) is kept constant and the concentration of added calcium is increased, phosphorylated oligos Similar results were obtained for both sodium and calcium sugar salts. Added calcium salt 6mM
When it became above, the effect by addition of phosphorylated oligosaccharide fell.

(Influence on remineralization effect with various Ca / P using phosphorylated oligosaccharide)
The concentration ratio of calcium and phosphorus was changed as shown in Table 3 below, and incubated at 37 ° C. for 17.5 hours or 1 week using the above simple test system (however, in Table 3,
P / Ca).

この結果を図６Ａ〜Ｃに示す（縦軸に再石灰化促進率（％）を示し、横軸にＰ／Ｃａを
示す）。図６Ａは、リン酸化オリゴ糖塩無添加のコントロールの結果を示し、白四角は１
７．５時間処理を、黒菱形は、１週間処理を表す。図６Ｂは、リン酸化オリゴ糖ナトリウ
ム塩の結果を示し、白三角は１７．５時間処理を、黒三角は、１週間処理を表す。図６Ｃ
は、リン酸化オリゴ糖カルシウム塩の結果を示し、白丸は１７．５時間処理を、黒丸は、
１週間処理を表す。図６Ａ〜Ｃに示したように、Ｃａを１．５ｍＭと一定にして、リン濃
度を変化させてＰ／Ｃａ比を変化させた場合、リン酸化オリゴ糖のナトリウム塩およびカ
ルシウム塩ともに、比較的効果的に再石灰化が生じると考えられた。本結果により、カル
シウム塩の方が高濃度のリンにおいても安定しているとも考えられる。

The results are shown in FIGS. 6A to 6C (the vertical axis indicates the remineralization promotion rate (%), and the horizontal axis indicates P / Ca). FIG. 6A shows the results of the control without addition of phosphorylated oligosaccharide salt.
7.5 hour treatment, black diamond represents 1 week treatment. FIG. 6B shows the results for phosphorylated oligosaccharide sodium salt, the white triangle represents the 17.5 hour treatment and the black triangle represents the one week treatment. 6C.
Shows the result of phosphorylated oligosaccharide calcium salt, white circle is treated for 17.5 hours, black circle is
Represents 1 week treatment. As shown in FIGS. 6A to 6C, when the P / Ca ratio was changed by changing the phosphorus concentration while keeping Ca constant at 1.5 mM, both the sodium salt and calcium salt of the phosphorylated oligosaccharide were relatively It was thought that remineralization would occur effectively. From this result, it is considered that the calcium salt is more stable even at a high concentration of phosphorus.

(Example 5)
This example shows a comparison of the effects of phosphorylated oligosaccharides and other anti-cariogenic substances on remineralization. Xylose, xylitol, palatinose, and palatinit were used as anti-cariogenic substances. In order to examine the effect on remineralization, the simplified system in Example 4 was used. In a simple system, incubation was performed at 37 ° C. for 8 days. These results are shown in FIGS. 7A to 7C (the vertical axis indicates the remineralization acceleration rate (%), and the horizontal axis indicates the sample concentration (%)).
. FIG. 7A shows the results of phosphorylated oligosaccharide salt, black triangles represent calcium salts and white triangles represent sodium salts. FIG. 7B shows the results for xylitol, black circles represent xylitol and white circles represent xylose. FIG. 7C shows the results for palatinit, where black squares represent paratinite and white squares represent palatinose. According to FIGS. 7A-C, for phosphorylated oligosaccharides, 0.1
Other anti-cariogenic substances (xylitol, palatinose, and palatinit) have been reported previously (Japanese Patent Laid-Open No. 2000-128752;
As shown in Japanese Patent Application Laid-Open No. 2000-247852 and the like, the remineralization effect was shown at an addition concentration of 20% (xylose has a low remineralization percentage at any concentration).

(Example 6)
This example shows the verification of the decalcification inhibitory effect of phosphorylated oligosaccharides.

A decalcification solution having the following composition was prepared: 6.0 mM calcium chloride solution, 3.6 mM potassium dihydrogen phosphate, 2% lactic acid solution, and 5 mg / ml hydroxyapatite solution,
pH 5.0. 125 μl of this demineralized solution and 125 μl of a phosphorylated oligosaccharide sodium salt solution having a final concentration of 0.2% and 2% were stirred and incubated at 37 ° C. for 2 days. Subsequently, the mixture was centrifuged at 12,000 rpm for 3 minutes, and the calcium concentration of the resulting supernatant was determined by OCP.
Measurement was made using the C method. The calcium concentration at the time of addition was compared with the calcium concentration after treatment. The difference in calcium concentration after treatment from the calcium concentration at the time of addition in the presence of the test sample (
A test sample was considered to have a demineralization-inhibiting effect if it was low compared to a control (without the test sample). 0.2% compared to control without phosphorylated oligosaccharide (5 mM)
And 2% phosphorylated oligosaccharide sodium salt solutions were both low (3 mM and 2 m
M), therefore, phosphorylated oligosaccharide sodium salt was considered to have a decalcification inhibitory effect.

(Example 7)
This example shows a synergistic effect of phosphorylated oligosaccharide with fluorine on the remineralization effect.

With the composition shown in Table 4 below, the remineralization effect in the presence or absence of phosphorylated oligosaccharide was verified.

再石灰化に対する効果を調べるために、実施例４における簡易系を用いた。簡易系にお
いて、３７℃で５日間のインキュベートを行った。次いで、可溶性カルシウムの量をＯＣ
ＰＣ法を用いて測定した。薄層クロマトグラフィー（ＴＬＣ）によって、リン酸化オリゴ
糖の定性的確認を行った。ＴＬＣ分析条件は、以下の通りである：シリカゲルプレート（
Ｍｅｒｃｋ社製）、エタノール／脱イオン水／酢酸＝７０／３０／２、１回室温展開、サ
ンプル添加量５μｌ、マーカーとして、１％リン酸化オリゴ糖１μｌおよび１％マルトト
ライオース１μｌを使用。

In order to examine the effect on remineralization, the simplified system in Example 4 was used. In a simple system, incubation was performed at 37 ° C. for 5 days. The amount of soluble calcium is then
Measurement was made using the PC method. Qualitative confirmation of phosphorylated oligosaccharides was performed by thin layer chromatography (TLC). TLC analysis conditions are as follows: silica gel plate (
(Merck), ethanol / deionized water / acetic acid = 70/30/2, developed at room temperature once, sample addition amount 5 μl, 1 μl of 1% phosphorylated oligosaccharide and 1 μl of 1% maltotriose were used as markers.

The result of TLC analysis is shown in FIG. In FIG. 8, various concentrations (ppm) of fluorine are shown in each lane, the upper spot represents maltotriose, and the lower spot represents phosphorylated oligosaccharide. FIG. 9 shows the synergistic effect in remineralization of phosphorylated oligosaccharides and fluorine (
The vertical axis shows the remineralization acceleration rate (%), the horizontal axis shows the fluorine concentration (ppm); at each value, the left bar shows the control of the group without added phosphorylated oligosaccharide, and the right bar Is 0.
2% phosphorylated oligosaccharide added group is shown). Because fluorine is a highly reactive halogen element,
The effects of fluorine on phosphorylated oligosaccharides and calcium determination were investigated. Under the conditions of this experiment, the effect of the addition of fluorine seemed to be no problem (FIG. 8). C with addition of fluorine
Since the concentration ratio balance between a and P, which is difficult to insolubilize, is lost, the remineralization rate is reduced by the increase in fluorine concentration. However, when 0.2% phosphorylated oligosaccharide sodium salt was added, the remineralization effect tended to increase rather, and a remarkable synergistic effect could be confirmed (FIG. 9).

(Example 8)
This example shows the results of blending phosphorylated oligosaccharides into chewing gum and elution into the human oral cavity.

A plate gum (chewing gum on a plate; the weight per plate gum is about 3.2 g) containing a calcium salt of phosphorylated oligosaccharide (calcium content: 3.2%) shown in Table 5 below was prepared. .

本ガムを咀嚼した際の経時的な口腔内への溶出量を薄層クロマトグラフィー（ＴＬＣ）
で分析した。ＴＬＣ条件は、以下の通りである：展開プレート、シリカゲルプレート；展
開溶媒、エタノール／脱イオン水／酢酸＝７０／３０／２、展開温度、室温で１回展開；
スポットサンプル量、３μｌ；検出、検出液（硫酸／エタノール＝１：１）をプレートへ
噴霧した後、１３０℃で３分間処理することで、スポットが発色する。

Thin layer chromatography (TLC) shows the amount of elution into the oral cavity over time when chewing the gum
Analyzed with TLC conditions are as follows: development plate, silica gel plate; development solvent, ethanol / deionized water / acetic acid = 70/30/2, development temperature, development once at room temperature;
Spot sample amount, 3 μl; detection, detection solution (sulfuric acid / ethanol = 1: 1) is sprayed on the plate and then treated at 130 ° C. for 3 minutes, so that the spot develops color.

FIG. 10 shows the results of TLC analysis of phosphorylated oligosaccharides at standard solution concentrations. Each lane shows elution of various concentrations of phosphorylated oligosaccharide (1% xylitol is shown as the control on the left and 1% maltotriose (G3) is shown on the right), and the lower spot contains the phosphorylated oligosaccharide. The upper spot represents xylitol and maltotriose. FIG. 11 shows the elution amount over time when the phosphorylated oligosaccharide-containing gum is eaten. Each lane shows the elution of chewing time (1% phosphorylated oligosaccharide as the control on the left side, 1% xylitol and maltotriose (G3) on the right side), and the lower spot represents the phosphorylated oligosaccharide. The upper spot shows xylitol and maltotriose. This phosphorylated oligosaccharide is not degraded by salivary amylase. From both these figures, about 10 after the start of chewing
In minutes, phosphorylated oligosaccharides are present in the oral cavity at a relatively high concentration, and after 20 minutes 0.2%
It can be seen that it remains at a concentration of about 5%.

Example 9
This example demonstrates the effect of phosphorylated oligosaccharides on sucrose fermentation.

S. Mutans 8148 strain was cultured at 37 ° C. for 14 hours in 1,000 ml of brain heart infusion medium (manufactured by DIFCO). Next, the cells were 6,000 rpm
And collected by centrifugation for 20 minutes. The cells were washed with phosphate buffered saline (PBS, pH 7.2).
) And then suspended in the same PBS to 40% (v / v). The reaction mixture (250 μl) for the pH measurement test was obtained by adding 125 μl of 40% cell suspension and 80 mM sucrose 62.
. 5 μl, various oligosaccharides (5% phosphorylated oligosaccharide sodium and calcium salts)
Consists of 62.5 μl of aqueous solution. While incubating at 37 ° C., the pH of the reaction mixture was continuously measured with a Toa radio wave pH meter.

S. 0.684% sucrose or 0 in 20% cell suspension of mutans 8158 strain
. When 684% glucose is added, the pH of the reaction solution becomes 5.0 or less within 5 minutes.
It dropped to almost 4.0 in minutes. 5% phosphorylated oligosaccharide (PO-1 and PO-2)
Co-existence of the pH was clearly suppressed in all cases (data not shown). 5% PO
When the sodium salt and calcium salt were added, the pH drop caused by the fermentation of 0.684% sucrose was effectively suppressed (data not shown).

(Example 10)
In this example, a sugar alcohol of phosphorylated oligosaccharide was prepared.

10 g each of the PO-1 fraction having one phosphate group in the molecule of the phosphorylated oligosaccharide and the PO-2 fraction having two phosphate groups were dissolved in 100 ml of distilled water. Using a 1N sodium hydroxide solution to make a slightly alkaline solution having a pH of about 8, 30 ml of a 3% sodium borohydride solution was added to 100 ml of this solution, and then allowed to stand at 40 ° C. for 1 hour to reduce oligosaccharides. did. By the above method, the reducing end of the phosphorylated oligosaccharide was hydrogenated. After adjusting the pH of this hydrogenated solution to 7.5 with 1N hydrochloric acid solution and terminating the reaction, the 0.22 μm membrane permeation solution was desalted with an electrodialyzer (Asahi Kasei Micro Acylizer G3, AC210-400 membrane). , Strong cation exchange resin (Nisshin Kasei Dowex50w, 20-50MESH, H-Fo
rm) to obtain a sugar solution having a pH of 2.7. This solution was neutralized with 1N sodium hydroxide solution or calcium hydroxide solution, and then lyophilized to prepare a sodium salt or calcium salt of a phosphorylated oligosaccharide sugar alcohol.

Example 11
In this example, chondroitin sulfate oligosaccharide (unsaturated disaccharide (dimer)) was prepared.

4.8 g of sodium chondroitin sulfate (C type; manufactured by Katayama Chemical) was dissolved in 500 ml of distilled water (pH 6.0), and 15 U of chondroitinase ACII (Arthrobact)
er aurescens (manufactured by Seikagaku Corporation) was added and reacted at 37 ° C. for 23 hours. The reaction was stopped in a boiling bath and desalted as in Example 10 to prepare a sodium or calcium salt of chondroitin sulfate oligosaccharide.

Example 12
In this example, the remineralization effect of various substances was examined.

The remineralization simple test system of Example 4 was used, and the substances shown in Table 6 below were used as samples. All materials were prepared at a final concentration of 0.1%.

上記の表６中、番号１のＰＯｓＮａはリン酸化オリゴ糖（ＰＯ−１画分）のナトリウム
塩であり、番号２のＰＯ−２Ｎａはリン酸化オリゴ糖（ＰＯ−２画分）のナトリウム塩で
あり、番号３のＰＯｓＨ Ｎａはリン酸化オリゴ糖（ＰＯ−１画分）の糖アルコールのナ
トリウム塩であり、番号４のＰＯ−２Ｈ Ｎａはリン酸化オリゴ糖（ＰＯ−２画分）の糖
アルコールのナトリウム塩であり、番号５のＧ３はマルトトライオースであり、番号６の
Ｇｌｃ−６−Ｐはグルコース−６−リン酸であり、番号７のＳｅｒ−Ｐは、セリンリン酸
であり、番号８はコンドロイチン硫酸Ｃであり、番号９は、オリゴガラクツロン酸であり
、番号１０は、コンドロイチン硫酸の不飽和二糖（表６および図１２中、Ｄｉｍｅｒ Ｎ
ａ）であり、番号１１のＤ．Ｗ．は脱イオン水である。

In Table 6 above, POsNa number 1 is the sodium salt of phosphorylated oligosaccharide (PO-1 fraction), PO-2Na number 2 is the sodium salt of phosphorylated oligosaccharide (PO-2 fraction). Yes, POsH Na of number 3 is a sodium salt of sugar alcohol of phosphorylated oligosaccharide (PO-1 fraction), PO-2H Na of number 4 is sugar alcohol of phosphorylated oligosaccharide (PO-2 fraction) No. 5 G3 is maltotriose, No. 6 Glc-6-P is glucose-6-phosphate, No. 7 Ser-P is serine phosphate, No. 8 Is chondroitin sulfate C, number 9 is oligogalacturonic acid, number 10 is an unsaturated disaccharide of chondroitin sulfate (in Table 6 and FIG. 12, Dimer N
a) and D. W. Is deionized water.

The results are shown in FIG. 12 (the vertical axis indicates the remineralization acceleration rate (%), and the horizontal axis indicates the substance used).
. In the figure, those having a higher remineralization ratio than the result with deionized water were judged to have a remineralization effect. Sodium salt of phosphorylated oligosaccharide alcohol, glucose-6-phosphate, sodium salt of chondroitin sulfate C, sodium salt of unsaturated disaccharide of chondroitin sulfate was superior to or better than phosphorylated oligosaccharide sodium salt Remineralization effect was shown.

(Example 13)
In this example, the remineralization effect of various substances was examined.

The remineralization simple test system of Example 4 was used, and the substances shown in Table 7 below were used as samples.

表７中、番号１のＰＯｓＮａはリン酸化オリゴ糖（ＰＯ−１画分）のナトリウム塩（最
終濃度０．２％）であり、番号２のＰＯｓＮａはリン酸化オリゴ糖（ＰＯ−１画分）のナ
トリウム塩（最終濃度２．０％）であり、番号３はパラチノース（最終濃度２．０％）で
あり、番号４はパラチノース（最終濃度２０％）であり、番号５はキシリトール（Ｗａｋ
ｏ２４４−００５２）（最終濃度２％）であり、番号６はキシリトール（Ｗａｋｏ２４４
−００５２）（最終濃度２０％）であり、番号７はトレハロース（Ｗａｋｏ ０２２５２
）（最終濃度２％）であり、番号８はトレハロース（Ｗａｋｏ ０２２５２）（最終濃度
２０％）であり、番号９はソルビトール（Ｋａｔａｙａｍａ２８−４７７０）（最終濃度
２％）であり、番号１０はソルビトール（Ｋａｔａｙａｍａ２８−４７７０）（最終濃度
２０％）であり、番号１１のＧ３はマルトトライオース（最終濃度２％）であり、番号１
２のＤ．Ｗ．は脱イオン水（コントロール）であり、番号１３は有機酸（酒石酸）（最終
濃度０．２％）であり、番号１４は有機酸（酒石酸）（最終濃度１．４％）であり、番号
１５はデキストラン硫酸（最終濃度０．２％）である。

In Table 7, POsNa of No. 1 is a sodium salt (final concentration 0.2%) of phosphorylated oligosaccharide (PO-1 fraction), and POsNa of No. 2 is phosphorylated oligosaccharide (PO-1 fraction) Sodium salt (final concentration 2.0%), number 3 is palatinose (final concentration 2.0%), number 4 is palatinose (final concentration 20%), number 5 is xylitol (Wak
o244-0052) (final concentration 2%), number 6 is xylitol (Wako244)
-0052) (final concentration 20%), number 7 is trehalose (Wako 02252)
) (Final concentration 2%), number 8 is trehalose (Wako 02252) (final concentration 20%), number 9 is sorbitol (Katayama 28-4770) (final concentration 2%), number 10 is sorbitol ( Katayama 28-4770) (final concentration 20%), G3 of number 11 is maltotriose (final concentration 2%), number 1
2 D.E. W. Is deionized water (control), number 13 is an organic acid (tartaric acid) (final concentration 0.2%), number 14 is an organic acid (tartaric acid) (final concentration 1.4%), number 15 Is dextran sulfate (final concentration 0.2%).

The results are shown in FIG. 13 (the vertical axis indicates the remineralization acceleration rate (%), and the horizontal axis indicates the substance used).
. In the 20% addition group of xylitol, palatinose, and sorbitol, the remineralization effect as previously reported (same as above) was confirmed. Moreover, the remineralization effect was confirmed also in the dermatan sulfate similarly to chondroitin sulfate. Organic acids were also as effective as phosphorylated oligosaccharides.

(Example 14)
This example shows verification of the caries prevention effect in the artificial oral cavity device.

S. sobrinus 6715 strain (brain heart infusion medium (DI
Culture medium, heart infusion liquid medium (manufactured by DIFCO)
, Sample solutions (each solution refrigerated during the test), each at 6 ml / hour / tube,
It was fed onto bovine teeth (about 5 × 5 mm) left in a thermostatic chamber (37 ° C.). P on the tooth surface over time
H was measured. The results are shown in FIG. 14 (the vertical axis indicates the change in pH and the horizontal axis indicates the elapsed time; the white circle is the addition of 1% sugar (GF) only, and the black triangle is 1% GF + 5% phosphorus. Represents the addition of oxidized oligosaccharides (POs)). 16 hours later, scrape plaque on teeth, 500n
Turbidity was measured in m. Further, the amount of insoluble glucan (WIG) formed was measured by the phenol sulfuric acid method. The strength of the teeth was measured with a hardness meter. It was shown as a value (ΔH) subtracted from the strength of the untreated product. The results are shown in Table 8 below.

１％ＧＦ（砂糖）では、有機酸が発生し、１０時間もするとｐＨが５．６以下になり、
プラーク内に保持されていることが明らかであった。プラークも十分に形成されていた。
歯も脱灰が進み、もろくなっていた。一方、１％ＧＦおよび５％リン酸化オリゴ糖を含む
溶液は、全くプラークが形成されず、ｐＨも低下しなかった。すなわち、う蝕菌定着を阻
害しプラークを形成させず、歯の脱灰が抑制され、歯の強度変化がなかった。この結果よ
り、リン酸化オリゴ糖のう蝕予防効果が明らかになった。この現象は、ヒト口腔内におい
ても同様に生じていることが考えられる。

With 1% GF (sugar), an organic acid is generated, and after 10 hours, the pH becomes 5.6 or less.
It was clear that it was retained in the plaque. Plaques were well formed.
The teeth were demineralized and became brittle. On the other hand, in the solution containing 1% GF and 5% phosphorylated oligosaccharide, no plaque was formed and the pH was not lowered. That is, caries colonization was inhibited and plaque was not formed, tooth decalcification was suppressed, and there was no change in tooth strength. From this result, the caries preventive effect of phosphorylated oligosaccharide was clarified. This phenomenon is considered to occur in the human oral cavity as well.

(Example 15)
This example shows the preparation of phosphorylated oligosaccharides from various starches.

The starches used in this example were rice starch (trade name: Better Friend, manufactured by Shimada Chemical) and tapioca starch (manufactured by Sanwa Starch).

100 g of starch was placed in 800-1000 ml of water, and bacteria B. richenfo
50 μl of rmis-derived starch liquefied α-amylase (BLA) 5000 U / ml (Fuctamirase, obtained from Hankyu Kyoei Bussan Co., Ltd., 1%) was added, and gelatinized in a water bath at 50 ° C. for 48 hours. Further, this gelatinized starch was mixed with 50 μl of BLA 5000 U / ml (Fuctamirase, obtained from Hankyu Kyoei Bussan Co., Ltd., 1%), pullulanase 200 U / m.
l (promozyme: obtained from Novo Nordisk) 50 μl, glucoamylase (
GA) (416 U / ml) (obtained from Toyobo), 50 μl, and incubated at 50 ° C. for 48 hours. This was centrifuged at 8,000 rpm for 20 minutes. The supernatant was subjected to an anion exchange resin (Chitopearl BCW2501; manufactured by Fujibo) equilibrated with 10 mM acetate buffer (pH 4.5). Thorough washing with the same buffer was performed to remove neutral sugars, followed by elution with the same buffer containing 0.5 M sodium chloride. Each elution fraction was concentrated using an evaporator, desalted and lyophilized to obtain phosphorylated oligosaccharide.

The phosphorylated oligosaccharide obtained as described above was again applied to an anion exchange resin column (Chitopearl BCW2501) equilibrated with 20 mM acetate buffer (pH 4.5). The column was sufficiently washed with the same buffer to remove neutral sugars. Fractions eluted with the same buffer containing 0.15 M sodium chloride and then with the same buffer containing 0.5 M sodium chloride were collected. As a result of analysis based on the above structure determination method, these fractions were desalted and freeze-dried, so that 3 or more and 5 or less α-1,4 glucose were bound from the 0.15 M sodium chloride elution fraction. Phosphorylated oligosaccharide (PO-1 fraction) in which one phosphate group is bound to glucan is obtained, and from 0.5M sodium chloride elution fraction, 2 or more and 8 or less glucose α-1,4 bonds As a result, phosphorylated oligosaccharide (PO-2 fraction) in which two or more phosphate groups were bonded to the obtained glucan was obtained. The structural analysis of the above phosphorylated oligosaccharide followed Example 1.

(Example 16)
This example shows that chewing gum containing phosphorylated oligosaccharides had the effect of promoting enamel remineralization in early caries.

Sugarless gum (containing 45% xylitol) and POsCa-free sugarless gum (47%) containing 2.5% of POs Ca derived from potato starch as an average content
. Two types of granulated gum (containing about 1.5 g / grain) having 5% xylitol were prepared by a conventional method. In addition, all the experimental reagents used were special grade reagents. In addition, content is a ratio with respect to the gum total weight, respectively.

An enamel block was prepared by cutting an experimental surface standardized with a diamond saw (manufactured by LUXO) using a crown enamel portion of bovine teeth as a tooth material. (7 × 7 × 3 mm) Six samples of these enamel blocks were used as a room temperature polymerization resin (UNIFAST Trad, GC
The whole was arranged into a plate having a size of 15 × 50 mm and a thickness of 7 mm. Next,
Smooth fresh enamel was exposed by polishing the surface with No. 800 water-resistant paper. On the other hand, the dentin surface side was embedded in advance with an impression compound (manufactured by GC). For one enamel block-embedded plate prepared in this way, 0.1 M lactic acid gel (6 wt% ca
rboxymethylcellulose, pH 5.0) in 100 ml for 4 weeks at 37 ° C
Artificial caries were generated by immersing under the above conditions.

In 17 healthy subjects, POs Ca-containing gum or 2 POs Ca-free gums (3.0 g) were chewed for 20 minutes. This test was conducted without informing the subject of the gum type. 1 minute after starting chewing gum, then 1 to 3 minutes, 3 to 6 minutes, 6 to 10 minutes, 10 to 20 minutes 10 using
Aliquots were collected into ml plastic test tubes. About these saliva, saliva volume, saliva p
After immediately measuring the H value, the saliva supernatant was diluted 10-fold with distilled water, and then the 0.45 μm membrane (Mi
LIPORE's filtrate was measured for the Ca and inorganic P contents by the OCPC method (calcium C
Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.) and the molybdic acid method.

In 12 healthy subjects, POs Ca-containing gum or 2 POs Ca-free gums (3.0 g) were chewed for 20 minutes. The gum type was performed face down on the subject. Whole saliva (saliva A) up to 10 minutes in the first half after the start of chewing gum and total saliva (saliva B) in the last 10 minutes were collected in a 50 ml plastic test tube using a plastic funnel. Immediately after measuring the saliva amount and pH value of these collected saliva, immediately after the enamel block embedding plate in which the artificial caries was generated was placed in a plastic container (10 ×
30 × 60 mm) was injected with 7 ml or more of each saliva. This amount is such that the enamel block embedding plate is sufficiently immersed. The plate was immersed in saliva A for 10 minutes, and then immersed in saliva B for 10 minutes. Thereafter, the plate was taken out, and the plate surface was thoroughly washed with distilled water. This immersion operation was performed at 37 ° C., and this series of operations was repeated four times a day. After the operation for 1 day, the enamel block-embedded plate was refrigerated at 100% humidity. In this test, fresh human saliva was collected every day in the same manner, and was performed continuously for 4 days.
In addition, about the saliva used for the test, after diluting 10 times with distilled water using a part of the supernatant, 0.
Filtration was performed through a 45 μm membrane (Millipore). Filtrate Ca and inorganic P
The content was quantified daily by the method described above.

Each tooth enamel after being immersed in human saliva is treated with a hard tissue cutter (Isom) after the immersion test.
et, Buhler, USA), cut out sections of about 500 &micro; m thickness,
The sections were polished to a thickness of about 200 &micro; m with a No. 800 waterproof paper, and microradiographs of each section were taken (PW-1830, Philips, The Net).
herlands). Shooting conditions are tube voltage 25 kV, tube current 25 mA, tube / subject distance 3
70 mm. Subsequently, the image quantification method of Inaba et al. (Eur. J. Oral. Sci. 105).
: 74-84, 1997), the demineralization depth (Ld, μm) and the amount of mineral loss ΔZ (v
ol% · μm).

In 17 healthy subjects, POs Ca-containing gum or 2 POs Ca-free gums (3.0 g) were each chewed for 20 minutes. In this case, the amount of saliva (FIG. 15; the horizontal axis indicates the chewing time of the gum, the vertical axis indicates the amount of saliva (ml)), the saliva pH value (FIG. 16; the horizontal axis indicates the time of chewing the gum, and the vertical axis indicates pH indicates), Ca content in saliva (FIG. 18; horizontal axis indicates gum chewing time, vertical axis indicates calcium content (mg)), and P content in saliva (FIG. 17;
The abscissa indicates the gum chewing time, and the ordinate indicates the phosphorus amount (mg) over time, and is expressed as an integrated value from the start. In addition, changes in the Ca / P ratio in saliva (FIG. 19; the horizontal axis indicates the chewing time of the gum and the vertical axis indicates the Ca / P ratio) were also calculated. In both figures, POs C
a-containing gum (+ POs Ca-containing gum) is square, POs Ca-free gum (-POs
Ca-containing gum) is indicated by diamonds.

As a result, no statistical difference depending on the gum type was obtained for saliva secretion (FIG. 15), pH change (FIG. 16), and P content change (FIG. 17). In chewing the gum for 20 minutes, about 30 ml of saliva was secreted, and the saliva pH rose from 7.0 at the beginning of gum chewing to about 7.5 after 5 minutes. The amount of P in saliva secreted by chewing gum was about 5 mg, and it was revealed that a sufficient amount for remineralization was present compared to the amount of Ca.
On the other hand, the Ca content at 20 minutes from the start of chewing is higher than that of POs Ca-free gum.
It was revealed that the a-containing gum was dissolved about 4 times more in saliva (FIG. 18). Since P was originally present in a certain amount in saliva (FIG. 16), the Ca / P ratio was also significantly higher (p <0.001) in the POs Ca-containing gum (FIG. 19). Further, in the above analysis results, no statistically significant difference due to gender was obtained.

Table 9 below shows the analysis results of saliva A and saliva B when 12 healthy subjects chew 2 POs Ca-containing gums or 2 POs Ca-free gums (3.0 g) for 20 minutes. Table 9 shows a comparison of saliva volume, pH, and mineral content.

いずれのガムにおいても唾液量は唾液Ａが唾液Ｂの約２倍量分泌されていた。ＰＯｓ Ｃ
ａ非含有ガムに比較してＰＯｓ Ｃａ含有ガムにおいて唾液ＡではＣａ含量に有意な差が
得られたが、唾液Ｂでは、その差は少なくなった。Ｐ量はガムによる差および唾液Ａおよ
びＢ間の差は認められなかった。そのため、Ｃａ／Ｐ比はＰＯｓ Ｃａ含有ガム咀嚼時の
唾液ＡにおいてＰＯｓ Ｃａ非含有ガム咀嚼群の約４倍高い値となった。

In any gum, the amount of saliva was secreted about twice as much as saliva A than saliva B. POs C
A significant difference was obtained in the Ca content in the saliva A in the POs Ca-containing gum compared with the non-containing gum, but the difference was reduced in the saliva B. There was no difference in the amount of P between gum and saliva A and B. Therefore, the Ca / P ratio was about 4 times higher in saliva A during chewing POs Ca-containing gum than in the POs Ca-free gum chewing group.

Subsequently, the results of evaluating the effect of promoting remineralization on each treated tooth in 12 people are shown in FIG. 20A (the vertical axis indicates the demineralization depth (Ld, μm)) and the figure as the demineralization depth and mineral loss amount, respectively. 20B (the vertical axis indicates the amount of mineral loss ΔZ (vol% · μm)). In both figures, the horizontal axis is blank, POs Ca-containing gum, POs
It shows in order of Ca non-containing gum. A recovery of decalcified enamel teeth was observed in POs Ca-containing gums, both significantly in demineralization depth (Ld) and mineral loss (ΔZ), compared to non-POs Ca-containing gums. That is, in the POs Ca-containing gum chewing group, a result of promoting remineralization was obtained (p <0.001).

(Example 17)
In this example, the enamel remineralization promoting effect of chewing gum containing phosphorylated oligosaccharide was shown in the human oral cavity.

In the same manner as in Example 16, two types of granulated gums (about 1.5 g / grain) were prepared, POs Ca-containing gum and POs Ca-free sugarless gum. In addition, all the experimental reagents used were special grade reagents.

An enamel disc (φ5mm; thickness 1.5mm) is prepared from the crown enamel part of the bovine tooth, and the surface of the tooth head on the buccal side is polished with # 800 water-resistant paper to expose a smooth fresh enamel. I let you. The enamel disc thus prepared caused artificial caries by being immersed in 100 ml of 0.1 M lactic acid solution (pH 5.0) for 3 days at 37 ° C. After decalcification, three enamel discs were mounted on the removable right palate plate in the upper right molar palate area.

In 12 healthy subjects (6 males, 6 females; average age: 21 years old), POs
Two pieces (3.0 g) of Ca-containing gum, POs Ca-free gum, or sucrose gum (including 62% sucrose) were chewed for two minutes (one tablet; about 1.5 g) at a time for 20 minutes. . This test was taken four times a day. The gum type was carried out with the tester and the subject lying down. Each gum group was run for 2 consecutive weeks, with a 1 week interval between each test group. The palate plate was attached for 20 minutes during chewing gum and after chewing. During the test period,
The subject avoided the use of a fluorinated agent, and stored the palate plate so as not to dry under the condition of 100% humidity except when wearing.

The mounting test teeth were collected from each subject's palate plate 1, 2, and 4 weeks later. About 200 & m thickness
icro; m sections were cut from each enamel and microradiographs of each section were taken (
PW-1830, Philips, The Netherlands). The shooting conditions were a tube voltage of 25 kV, a tube current of 25 mA, and a tube / subject distance of 370 mm. Next, the decalcification depth (ld, μm) was measured by the image quantification method of Inaba et al. (Eur. J. Oral. Sci. 105: 74-84, 1997). The ld value was defined in the mineral distribution profile as the distance from the dentition surface position to the lesion where the mineral content reached a level of 95% of the mineral content of healthy tissue. The remineralization rate was calculated as the ld value decrease rate (%) relative to the starting ld value after decalcification. The result of the remineralization rate is shown in FIG. In FIG. 21, the abscissa represents the sucrose gum group (Suc) in the order of 1 week, 2 weeks, and 4 weeks, respectively.
), POs Ca-free gum group (Xyl), and POs Ca-containing gum group (POs). The vertical axis represents the remineralization rate (%).

The remineralization rate of the POs Ca-containing gum group (POs) was 67%, 54%, and 76% after 1 week, 2 weeks, and 4 weeks, respectively. The POs Ca-free gum group (Xyl) was 12-23%, which was lower than the POs Ca-containing gum group. The sucrose gum group (Suc) showed a positive remineralization rate until 2 weeks later, but finally reached a negative value at 4 weeks, indicating decalcification.

Also in the human oral cavity test, a higher remineralization promoting effect was obtained in POs Ca-containing gum than in POs Ca-free gum and sucrose gum. That is, all 12 subjects ate all types of gums every 2 weeks, and significant results were obtained in POsCa-containing gums. From this, it was confirmed by a human oral test that a high remineralization promoting effect can be obtained by blending POs Ca into the gum. It was also confirmed in the oral cavity that taking POs Ca-containing gum products on a daily basis promoted remineralization of the initial caries and was very effective in preventing caries.

(Example 18)
In this example, component analysis of saliva secreted when a candy containing phosphorylated oligosaccharide was ingested was performed.

  Candy was prepared with the composition shown in Table 10 below.

健常成人４名を被検者として、キャンデー１個（４．７ｇ）を摂取した際の分泌唾液を採
取した。キャンデーは口腔内で摂取後およそ１０分間存在した。唾液の採取は、（１）０
〜１分間、（２）１〜３分間、（３）３〜６分間、（４）６〜１０分間の４つの時点に分
けて実施した。分泌唾液は、漏斗を通して１５ｍｌ試験管内に採取した。採取直後に分泌
唾液を攪拌し、唾液のｐＨおよび分泌量を測定した。この結果をそれぞれ図２２および２
３に示す。図２２は、横軸に摂取時間（分）および縦軸にｐＨを示す。口腔内の唾液ｐＨ
は７で一定していた。図２３は、横軸に摂取時間（分）および縦軸に唾液量（ｍｌ／分）
を示す。摂取時間の経過を通して唾液の分泌量に大きな変化はなかった。

With 4 healthy adults as subjects, the secretory saliva when one candy (4.7 g) was ingested was collected. The candy was present in the oral cavity for approximately 10 minutes after ingestion. Saliva collection is (1) 0
It was divided into four time points: ˜1 minute, (2) 1 to 3 minutes, (3) 3 to 6 minutes, and (4) 6 to 10 minutes. Secretory saliva was collected through a funnel into a 15 ml test tube. Immediately after collection, the secretory saliva was stirred, and the pH and secretion amount of the saliva were measured. The results are shown in FIGS. 22 and 2, respectively.
3 shows. FIG. 22 shows the intake time (minutes) on the horizontal axis and pH on the vertical axis. Saliva pH in the oral cavity
Was constant at 7. FIG. 23 shows the intake time (min) on the horizontal axis and the amount of saliva (ml / min) on the vertical axis.
Indicates. There was no significant change in salivary secretion over the course of the intake.

Then 1800 μl of saliva is placed in 4 centrifuge tubes, 200 μl of 1N HCl solution is added to each tube, mixed well, centrifuged at 10,000 × g for 3 minutes, and 0.
5 μm membrane treatment was performed. Of the obtained supernatant, 10 μl of calcium content was measured by the OCPC method, and 50 μl of phosphorus content was measured by the molybdenum method. The calcium content and phosphorus content are shown in FIG. FIG. 24 shows the intake time (min) on the horizontal axis, calcium or phosphorus content (mM) on the left vertical axis, and the Ca / P ratio on the right vertical axis. There was no significant change in the calcium content and phosphorus content throughout the intake time (maintained around 0.6).

Example 19
In this example, candy and soft candy blended with phosphorylated oligosaccharides were prepared, and the effect of promoting remineralization was examined.

In the formulation shown in Table 11 below, candy (4.7 g / piece) and soft candy (4
. 0 g / piece) was prepared according to a conventional method.

上記キャンデーおよびソフトキャンデーに蒸留水１０ｍｌを添加し、これを沸騰浴中で溶
解させた。得られた抽出溶液のｐＨを微量ｐＨメーターを用いて測定した。次いで１０，
０００×３分、遠心分離し、０．５μｍ膜処理を行った。得られた上清溶液のうち１０μ
ｌについてＯＣＰＣ法を用いてカルシウム濃度を測定し、５０μｌについてバナドモリブ
デン酸法を用いて無機リン濃度を測定した。この結果を表１２に示す。表１２は、ソフト
キャンデーおよびキャンデーの抽出物におけるＣａおよびＰのミネラル含量を示す。

10 ml of distilled water was added to the above candy and soft candy, and this was dissolved in a boiling bath. The pH of the obtained extraction solution was measured using a trace pH meter. Then 10,
Centrifugation was performed at 000 × 3 minutes, and 0.5 μm membrane treatment was performed. 10 μm of the obtained supernatant solution
For l, the calcium concentration was measured using the OCPC method, and for 50 μl, the inorganic phosphorus concentration was measured using the vanadomolybdic acid method. The results are shown in Table 12. Table 12 shows the Ca and P mineral content in soft candy and candy extracts.

さらに、上記表１２の分析結果より、この抽出溶液を２倍希釈および１０倍希釈した溶
液において、以下の表１３の組成にカルシウム、リン含量を補正した上で、ハイドロキシ
アパタイトの再石灰化促進効果を評価した。表１３は、再石灰化促進の評価系の組成を示
す。

Furthermore, from the analysis results of Table 12, the recalcification promoting effect of hydroxyapatite is obtained after correcting the calcium and phosphorus contents in the composition of Table 13 below in a solution obtained by diluting the extracted solution twice and 10 times. Evaluated. Table 13 shows the composition of the evaluation system for promoting remineralization.

この結果を図２５に示す。図２５は、縦軸に再石灰化促進率（％）、そして横軸には順に
ソフトキャンデー抽出溶液、キャンデー抽出溶液、および２％マルトオリゴ糖溶液の再石
灰化促進率の結果を示す。

The result is shown in FIG. FIG. 25 shows the results of the remineralization promotion rate (%) on the vertical axis and the remineralization promotion rate of the soft candy extraction solution, the candy extraction solution, and the 2% maltooligosaccharide solution in order on the horizontal axis.

For both candy and soft candy, a high remineralization promoting effect was observed at 10-fold dilution.

(Example 20)
Dentifrices having the compositions shown in Table 14 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 21)
Dentifrices having the compositions shown in Table 15 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 22)
Dentifrices having the compositions shown in Table 16 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 23)
Dentifrices having the compositions shown in Table 17 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 24)
Dentifrices having the compositions shown in Table 18 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 25)
Dentifrices having the compositions shown in Table 19 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 26)
Dentifrices having the compositions shown in Table 20 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 27)
Dentifrices having the compositions shown in Table 21 below were produced according to a conventional method. PO-Zn was prepared in the same manner as in Example 2 except that a 1N zinc hydroxide solution was used for neutralization.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 28)
Dentifrices having the compositions shown in Table 22 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 29)
Mouthwashes having the compositions shown in Table 23 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 30)
Mouthwashes having the compositions shown in Table 24 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 31)
Mouthwashes having the compositions shown in Table 25 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 32)
Mouthwashes having the compositions shown in Table 26 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 33)
Mouthwashes having the compositions shown in Table 27 below were produced according to a conventional method. POs Zn was prepared in the same manner as in Example 2 except that a 1N zinc hydroxide solution was used for neutralization.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 34)
An oral ointment having the composition shown in Table 28 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 35)
An oral ointment having the composition shown in Table 29 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 36)
Dentifrices having the compositions shown in Table 30 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 37)
Dentifrices having the compositions shown in Table 31 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 38)
Dentifrices having the compositions shown in Table 32 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 39)
Dentifrices having the compositions shown in Table 33 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 40)
Dentifrices having the compositions shown in Table 34 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 41)
Dentifrices having the compositions shown in Table 35 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 42)
Dentifrices having the compositions shown in Table 36 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 43)
Dentifrices having the compositions shown in Table 37 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 44)
Dentifrices having the compositions shown in Table 38 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 45)
Mouthwashes having the compositions shown in Table 39 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 46)
Mouthwashes having the compositions shown in Table 40 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 47)
Mouthwashes having the compositions shown in Table 41 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 48)
Mouthwashes having the compositions shown in Table 42 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 49)
Mouthwashes having the compositions shown in Table 43 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 50)
An oral ointment having the composition shown in Table 44 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 51)
An oral ointment having the composition shown in Table 45 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 52)
Dentifrices having the compositions shown in Table 46 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 53)
Dentifrices having the compositions shown in Table 47 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 54)
Dentifrices having the compositions shown in Table 48 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 55)
A dentifrice having the composition shown in Table 49 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 56)
Dentifrices having the compositions shown in Table 50 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 57)
Dentifrices having the compositions shown in Table 51 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 58)
Dentifrices having the compositions shown in Table 52 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 59)
Dentifrices having the compositions shown in Table 53 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 60)
Dentifrices having the compositions shown in Table 54 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 61)
A mouthwash having the composition shown in Table 55 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 62)
Mouthwashes having the compositions shown in Table 56 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 63)
Mouthwashes having the compositions shown in Table 57 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 64)
A mouthwash having the composition shown in Table 58 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 65)
A mouthwash having the composition shown in Table 59 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

Example 66
An oral ointment having the composition shown in Table 60 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 67)
An oral ointment having the composition shown in Table 61 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

Example 68
Dentifrices having the compositions shown in Table 62 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 69)
Dentifrices having the compositions shown in Table 63 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 70)
Dentifrices having the compositions shown in Table 64 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 71)
Dentifrices having the compositions shown in Table 65 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 72)
Dentifrices having the composition shown in Table 66 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 73)
Dentifrices having the compositions shown in Table 67 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 74)
Dentifrices having the compositions shown in Table 68 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 75)
Dentifrices having the compositions shown in Table 69 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 76)
Dentifrices having the compositions shown in Table 70 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 77)
Mouthwashes having the compositions shown in Table 71 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 78)
Mouthwashes having the compositions shown in Table 72 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 79)
Mouthwashes having the compositions shown in Table 73 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 80)
Mouthwashes having the compositions shown in Table 74 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 81)
A mouthwash having the composition shown in Table 75 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 82)
An oral ointment having the composition shown in Table 76 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 83)
An oral ointment having the composition shown in Table 77 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 84)
Dentifrices having the compositions shown in Table 78 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 85)
Dentifrices having the compositions shown in Table 79 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 86)
Dentifrices having the compositions shown in Table 80 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 87)
Dentifrices having the compositions shown in Table 81 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 88)
Dentifrices having the compositions shown in Table 82 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

Example 89
Dentifrices having the compositions shown in Table 83 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 90)
Dentifrices having the compositions shown in Table 84 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 91)
Dentifrices having the compositions shown in Table 85 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 92)
Dentifrices having the compositions shown in Table 86 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 93)
Mouthwashes having the compositions shown in Table 87 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 94)
Mouthwashes having the compositions shown in Table 88 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 95)
Mouthwashes having the compositions shown in Table 89 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

Example 96
Mouthwashes having the compositions shown in Table 90 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 97)
Mouthwashes having the compositions shown in Table 91 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 98)
An oral ointment having the composition shown in Table 92 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

Example 99
An oral ointment having the composition shown in Table 93 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 100)
Dentifrices having the compositions shown in Table 94 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 101)
Dentifrices having the compositions shown in Table 95 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 102)
Dentifrices having the compositions shown in Table 96 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 103)
Dentifrices having the compositions shown in Table 97 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 104)
Dentifrices having the compositions shown in Table 98 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 105)
Dentifrices having the compositions shown in Table 99 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 106)
Dentifrices having the compositions shown in Table 100 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 107)
Dentifrices having the compositions shown in Table 101 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 108)
Dentifrices having the compositions shown in Table 102 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 109)
Mouthwashes having the compositions shown in Table 103 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 110)
Mouthwashes having the compositions shown in Table 104 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 111)
Mouthwashes having the compositions shown in Table 105 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 112)
Mouthwashes having the compositions shown in Table 106 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 113)
Mouthwashes having the compositions shown in Table 107 below were produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 114)
An oral ointment having the composition shown in Table 108 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 115)
An oral ointment having the composition shown in Table 109 below was produced according to a conventional method.

これらの配合によれば、良好な抗う蝕機能が達成される。

According to these formulations, a good anti-cariogenic function is achieved.

(Example 116)
Artificial saliva having the composition shown in Table 110 below was produced according to a conventional method.

この唾液は、再石灰化促進効果に優れ、そして口腔内のｐＨを中性に戻すのに優れる。

This saliva is excellent in the effect of promoting remineralization and excellent in returning the pH in the oral cavity to neutral.

(Example 117)
Artificial saliva having the composition shown in Table 111 below was produced according to a conventional method.

この唾液は、再石灰化促進効果に優れ、そして口腔内のｐＨを中性に戻すのに優れる。

This saliva is excellent in the effect of promoting remineralization and excellent in returning the pH in the oral cavity to neutral.

Artificial saliva can be similarly adjusted by adding a buffer other than POs Ca and POs Na.

Claims (1)

A composition for eating and drinking having an anti-cariogenic function, wherein the composition contains a buffer having a pH buffering action in the oral cavity, but the buffer is not the following (1) or (2) Composition: (1) A glucan prepared from potato starch, consisting of 3-5 glucose with α-1,4 linkages, and having one phosphate group attached thereto, phosphorylated oligosaccharides; Or (2) a phosphorylated oligosaccharide, which is a glucan prepared from potato starch, consisting of 2-8 glucose with α-1,4 linkages, and 2 phosphate groups attached to it.

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