JP5925431B2 - Caries prevention agent - Google Patents

Description

  The present invention relates to a caries preventive agent comprising an algal extract of a millaceae, and in particular, a caries having an effect of suppressing adhesion and proliferation to plaques or biofilms in the oral cavity, comprising an extract of an algal extract of a myceae The present invention relates to a preventive agent, and a pharmaceutical composition, a food composition, a dentifrice and a mouthwash containing the preventive agent for caries.

  Oral bacteria grow on adhering to plaques or biofilms in the oral cavity, causing caries, ie caries, alveolar pyorrhea and other periodontal diseases. The plaque or biofilm grows when saliva biopolymers adhere to the tooth surface to form a pellicle film, and oral bacteria adhere to the pellicle. Therefore, in order to prevent dental caries and periodontal disease, it is necessary to suppress or prevent the adhesion of oral bacteria to the pellicle. Sugar alcohols such as erythritol and xylitol are known to have caries-inhibiting effects, but sugar alcohols do not produce lactic acid that causes caries due to metabolism of oral bacteria, making it difficult to produce caries. It is not involved in inhibiting or preventing oral bacterial adhesion.

  When oral bacteria adhere to the pellicles and other oral bacteria, it is considered that a lectin which is a sugar chain binding protein produced by oral bacteria is involved. That is, it is considered that oral bacteria adhere to the pellicle when the biopolymer of saliva stagnated in the pellicle, that is, the polysaccharide or glycoprotein, and the oral bacterial lectin perform sugar chain-specific binding. Then, by administering the lectin that competes with the oral bacterial lectin from the outside into the oral cavity and inhibiting the binding of the oral bacterial lectin to the sugar chain of the pellicle, the adhesion and growth of oral bacteria on the biofilm is suppressed. Alternatively, it can be inhibited to prevent oral bacterial infections such as tooth decay and periodontal disease.

  Non-patent documents 1 to 3 report lectins that actually inhibit the binding between oral bacteria and pellicles based on such an idea. The lectins reported in these documents include plant glucose / mannose recognition lectins and algal mucin recognition lectins. Patent Document 1 discloses that mushroom-derived lectin ABA and the like have an effect of suppressing adhesion and proliferation to plaque or biofilm in the oral cavity.

Teixeira, E .; H. Et al., J Appl. Microbiol. 101: 111-6 (2006). Teixeira, E .; H. Et al., J Appl. Microbiol. 103: 1001-6 (2007). Islam, B.I. Et al., J Appl. Microbiol. 106: 1682-9 (2009).

PCT / JP2009 / 069033 specification

  However, when a caries preventive agent is developed using mushroom extract components, the protein in the extract components requires several milligrams per milliliter, and it is difficult to stably prepare a large amount. In addition, mushroom extract components are not suitable for addition to foods due to their strong odor and puffy taste. Therefore, it is necessary to develop a caries preventive agent derived from a material whose odor and taste do not matter.

  The present invention provides a preventive agent for dental caries comprising an algal extract of Myraceae algae.

  In the caries preventive agent of the present invention, the algae may belong to the genus Mill.

  In the caries preventive agent of the present invention, the algae may belong to at least one species of the genus Mir selected from the group consisting of mill, isemomil, tamamil, cobusimil, chromyl, mottle mill, and nagamil.

  In the caries preventive agent of the present invention, the algae may be a mill.

  In the caries preventive agent of the present invention, the algal extract may be obtained by extracting a crushed algal extract with an aqueous solution.

  In the caries preventive agent of the present invention, the prealgal extract may be a lectin that binds to a sugar chain that competes with GalNAc.

  The present invention provides a pharmaceutical composition comprising the caries preventive agent of the present invention.

  The present invention provides a food composition containing the agent for preventing dental caries of the present invention.

  The present invention provides a dentifrice containing the caries preventive agent of the present invention.

  The present invention provides a mouth rinse containing the caries preventive agent of the present invention.

  The present invention provides a method for preventing caries comprising the step of administering the agent for preventing caries of the present invention into the oral cavity.

  The algae of the present invention belong to the family Chlorophyceae, Coleaceae of the order of the order Milly (Codiales, which is sometimes referred to as Bryopsisales). Codium fragile, Icemomil (Codium tomentosum), Tamil (Codium minus), Cobum smirgoum (Codium subtubulosum), Codrum intumaum, Codium intricaum, Codrum Codeumactum, Codium barbatum, Sa Butmir (Codium contractum), including Hiramiru (Codium latum) and Nagamiru (Codium cylindricum), not limited to and may belong to the species codium (Codium).

  The method for recovering the algal extract of the present invention may be any method as long as the activity of suppressing or inhibiting the binding between oral bacteria and the biopolymer of saliva is not substantially impaired.

  The algal extract of the present invention may be collected from any tissue of algal bodies. The algal bodies may be subjected to an extraction treatment after washing, after being crushed with a homogenizer or the like, or powdered by lyophilization, or powdered with a pulverizer. The aqueous solution used for extraction contains an aqueous solution containing NaCl or other salts such as physiological saline, acetone or other water-soluble organic solvent and / or a mixture of alcohol and water, and purified water or deionized water. However, other aqueous solvents well known to those skilled in the art can be used without being limited thereto. Here, the alcohol includes ethanol, ethylene glycol, butylene glycol, and glycerin, but may be an alcohol that is not limited thereto. The aqueous solvent may have an acidic or alkaline pH. In order to adjust the pH of the aqueous solvent, adipic acid, citric acid, gluconic acid, succinic acid, acetic acid, tartaric acid, lactic acid, fumaric acid, malic acid, phosphoric acid, potassium carbonate, sodium carbonate, sodium bicarbonate, phosphorus A pH adjuster may be used, including but not limited to the potassium or sodium salts of acid salts. In order to keep the pH constant, buffers such as, but not limited to, Tris hydrochloric acid buffer, Hepes buffer, phosphate buffer, acetic acid buffer, citrate buffer, and glycine-hydrochloric acid buffer may be used. Extraction methods include, but are not limited to, immersion extraction methods, pressurized extraction methods, and supercritical or subsupercritical extraction methods. The extraction condition may be any condition as long as it does not substantially impair the activity of suppressing or inhibiting the binding between oral bacteria and the biomolecules of saliva. The extraction time is from 10 minutes to 24 hours. The extraction temperature may be 4 ° C. or higher, preferably room temperature or higher, and more preferably 15 ° C. or higher.

  Purification and concentration of the algae extract include, but are not limited to, centrifugation, salting out, dialysis, vacuum concentration, ultrafiltration, gel filtration, ion exchange chromatography, affinity chromatography, etc. It may be performed by using alone or in any combination.

  After purification or concentration, if necessary, the algal extract may be dried under reduced pressure or diluted with the solvent. Moreover, the said extract may be mixed and used.

  In this specification, a lectin refers to a protein other than an antibody, T cell receptor, Toll-like receptor or other immune protein involved in the immune system of an animal and having the ability to specifically bind to a sugar chain. . Normally, lectins can aggregate red blood cells and other animal cells. According to Horikiru Osamu (Chemistry and Biology, 32: 586-594, (1994)), there is a common point that a lectin derived from an algae of the family Mill binds to a sugar chain that competes with GalNac.

  The extract or lectin of the present invention can measure the hemagglutination activity by a method well known to those skilled in the art using blood of humans, pigs, sheep, rabbits and the like. When erythrocytes are significantly aggregated by adding the test solution, it can be estimated that the lectin is contained in the test solution. In addition, the structure of the sugar chain to which the lectin binds can be examined by examining whether or not erythrocyte aggregation is inhibited at a low concentration by adding various sugars to the test solution.

The lectin of the present invention binds to a sugar chain that competes with GalNAc. This suggests that the end of the sugar chain to which the lectin of the present invention binds is GalNAc. However, the lectin of the present invention does not necessarily bind to all sugar chains having GalNAc at the terminal. The second to third sugar residues from the end bind only to a sugar chain having a specific structure, but the binding may be inhibited in the presence of GalNAc.

  The lectin contained in the dental caries preventive agent of the present invention may be a protein purified from an algal extract of a mysid algae, a protein expressed and purified in various hosts by recombinant DNA technology, or the alga or Does the extract have sufficient activity to suppress or inhibit the adhesion and growth of oral bacteria, especially mutans bacteria, to the oral hard tissue when administered into the oral cavity? There are cases. That is, the lectin may be used as an algal body of a mycelia or a processed product thereof.

  The dosage form of the dental caries preventive agent of the present invention may be any dosage form as long as it does not substantially impair the activity of inhibiting or inhibiting the binding between the oral bacteria of the present invention and the biopolymer of saliva. Absent.

  The pharmaceutical composition of the present invention includes all pharmaceuticals for suppressing or inhibiting the adhesion and growth of oral bacteria, particularly mutans bacteria, to the oral hard tissue, and the oral bacteria and saliva biopolymers of the present invention. Any other component may be included on the condition that the activity of suppressing or inhibiting the binding to is not substantially impaired.

  The food composition of the present invention may contain any other components provided that it does not substantially impair the activity of suppressing or inhibiting the binding between the oral bacteria of the present invention and the biomolecules of saliva. It doesn't matter. The food composition of the present invention may contain sugar alcohols such as erythritol and xylitol.

  Dentifrices of the present invention include, but are not limited to, toothpastes, liquid toothpastes, and toothpastes. The mouth washes of the present invention include, but are not limited to, mouthwash, gargle tablets, troches and chewing gum.

  The dentifrice or mouth rinse of the present invention contains any other component, provided that it does not substantially impair the activity of inhibiting or inhibiting the binding of the oral bacteria of the present invention to the biomolecules of saliva. It does not matter. Specific examples of the components include, but are not limited to, excipients, emulsifiers, stabilizers, fragrances, and sweeteners.

  In the case of a dentifrice, for example, abrasives, binders, thickeners, surfactants, sweeteners, preservatives, fragrances, colorants, various active ingredients, and the like can be blended. It can manufacture by mixing with. In addition, the compounding quantity of these components can be made into a normal quantity in the range which does not impair the activity which suppresses or inhibits the coupling | bonding of the oral bacteria of this invention and the biopolymer of saliva.

  Abrasives include silica-based abrasives such as precipitated silica, silica gel, aluminosilicate, zirconosilicate, dicalcium phosphate dihydrate and anhydrous, calcium pyrophosphate, zeolite, zirconium silicate, synthetic resin-based abrasive Etc., but not limited to

  Binders include, but are not limited to, sodium carboxymethylcellulose, hydroxyethylcellulose, carrageenan, gelatin and the like. Thickeners include, but are not limited to glycerin and sorbit.

  As the surfactant, an anionic surfactant, a cationic surfactant, a nonionic surfactant, and the like can be blended, and specifically, sodium lauryl sulfate is included, but not limited thereto.

  Sweetening agents include, but are not limited to, saccharin sodium, stevioside, stevia extract.

  Preservatives include, but are not limited to, paraoxybenzoic acid esters and sodium benzoate. Fragrances include, but are not limited to, 1-menthol.

  Colorants include, but are not limited to Blue No. 1 and Yellow No. 4.

  Various active ingredients include, but are not limited to, sodium fluoride, potassium fluoride, ammonium fluoride, and the like.

  The algal extract of the mycelial algae of the present invention comprises an aqueous solution containing NaCl or other salts such as physiological saline, a mixture of acetone or other water-soluble organic solvent and / or alcohol and water, purified water or It can be dissolved in other aqueous solvents well known to those skilled in the art including, but not limited to, deionized water. Here, the alcohol includes ethanol, ethylene glycol, butylene glycol, and glycerin, but may be an alcohol that is not limited thereto. The aqueous solvent may have an acidic or alkaline pH. In order to adjust the pH of the aqueous solvent, adipic acid, citric acid, gluconic acid, succinic acid, acetic acid, tartaric acid, lactic acid, fumaric acid, malic acid, phosphoric acid, potassium carbonate, sodium carbonate, sodium bicarbonate, phosphorus A pH adjuster may be used, including but not limited to the potassium or sodium salts of acid salts. In order to keep the pH constant, buffers such as, but not limited to, Tris hydrochloric acid buffer, Hepes buffer, phosphate buffer, acetic acid buffer, citrate buffer, and glycine-hydrochloric acid buffer may be used. The protein concentration of the extract of the present invention may be any concentration as long as it does not substantially impair the activity of inhibiting or inhibiting the binding between oral bacteria and saliva biopolymers. 25 μg / mL or more is preferable, and 125 μg / mL or more is more preferable. The lectin concentration of the present invention may be any concentration as long as it does not substantially impair the activity of suppressing or inhibiting the binding between oral bacteria and salivary biopolymers, but is 0.5 μg / mL. The above is preferable, and 5 μg / mL or more is more preferable.

The graph which shows the inhibitory effect by the extract of a mill and a willow matsutake with respect to adhesion | attachment and proliferation of the mutans bacteria to the substrate coated with saliva. The graph which shows the inhibitory effect by the refinement | purification lectin derived from a mill and a willow matsutake with respect to adhesion and proliferation of a mutans bacterium. The graph which shows the relationship between the protein concentration of the extract derived from the mill of another Example, and the inhibitory effect with respect to adhesion and proliferation of a mutans bacterium.

  The embodiments of the present invention described below are for illustrative purposes only and are not intended to limit the technical scope of the present invention. The technical scope of the present invention is limited only by the appended claims. Modifications of the present invention, for example, addition, deletion, and replacement of the configuration requirements of the present invention can be made on the condition that the gist of the present invention is not deviated.

  All documents mentioned herein are hereby incorporated by reference in their entirety.

Suppressing effect on adhesion and growth of mutans bacteria of extract components of mill and willow matsutake Extract preparations As raw materials, alga bodies of millet (Codyum fragile) and fruit bodies of willow matsutake (Agrocybe cylindracea) were used. The raw materials were purchased from growers in the state of raw seaweed and mushrooms that were not dried. A 10-fold amount of Dulbecco phosphate buffered saline (pH 7.3, D5652, Sigma-Aldrich, hereinafter referred to as “PBS”) is added to 50 to 100 g of the raw material, and a 0.5 mm diameter bead and a bead-type tissue Using a crushing apparatus (Bead-Beater, 1107900, Biospec Product), the mixture was crushed at 5000 rpm for 2 minutes until it became a paste. Thereafter, the mixture was allowed to stand at room temperature for 30 minutes, and an extract of mill and willow matsutake was prepared. The extract was centrifuged at 37,000 × g for 15 minutes, and the supernatant was collected. The protein concentration of the extract was measured using a BCA protein assay kit (Thermo Fisher Scientific Co., Ltd.). A two-fold dilution series of the extract with PBS was prepared and subjected to an experiment to evaluate the effect on the adhesion and growth of mutans bacteria.

The mutans bacteria were cultured overnight at 37 ° C. in Todd-Hewitt Broth, and the supernatant was removed by centrifugation at 1,000 × g for 10 minutes and then suspended in an equal volume of PBS. The same operation was repeated twice and washed, and then the absorbance was measured at 660 nm. Based on the measured absorbance, the mutans bacterium was diluted with PBS so that the OD ₆₆₀ was 0.05 or 0.1, and a mutans bacterium sample solution was prepared. The sample solution was sealed and used after being kept at 25 ° C. for 5 hours.

Biofilm suppression assay of extract 100 μL of human saliva for 3 hours after brushing was added to a multiplate (flat bottom, 96-well, polystyrene, Nunc Multisoap code: 467140) and coated at 37 ° C for 2 hours. It was. In the experiment, human saliva from three different subjects was used. Thereafter, the multi-plate was washed with PBS, and an extract of mill and willow matsutake diluted with PBS was added and incubated at 37 ° C. for 1 hour. The multiplate was then washed with PBS and 5 × 10 ⁷ Streptococcus mutans in 100 μL PBS was added to each well and incubated at 37 ° C. overnight. The plate was washed with PBS, 100 μL of 0.25% glutaraldehyde / PBS was added, and fixed at room temperature for 30 minutes. After washing with PBS, 100 μL of 2.3% crystal violet (for Gram stain) was added and stained at room temperature for 1 hour. Thereafter, it was washed with running water, 100 μL of ethanol was added, and the dye was eluted for 1 hour at room temperature. The number of bacteria was quantified by measuring the dye concentration in the ethanol solution of each well at an absorption wavelength of 570 nm using a plate reader.

Activity of suppressing or inhibiting the binding of biopolymers oral bacteria and saliva were quantified as inhibition index I _x. That is, the average A _x of the measured values of absorbance of each of the same conditions, the average A _n of measurements of absorbance of the wells was not performed coating with saliva for different subjects, they were subjected to coating by saliva for different subjects , from the average a _p of measurements of absorbance of the wells was not performed incubation with lectin, suppression index I _x of each of the same conditions is calculated using the following formula (I), optionally in the plot.

Formula (I) I _x = 100 × (A _x −A _n ) / (A _p −A _n )

A smaller value of the inhibition index _Ix indicates that the adhesion and growth of S. mutans was inhibited by the extract or purified lectin.

Assay Results FIG. 1 is a graph showing the inhibitory effect of Mill and Willow matsutake extracts on the adhesion and growth of S. mutans on saliva-coated substrates. The error bar for each experimental condition indicates the standard deviation of the measured value of the experimental result repeated twice for each identical condition. The four bar graphs from the right end of FIG. 1 are the results of the control experiment. In order from the right end, lectin-, saliva + and mutans bacteria + (saliva + mutans bacterium (positive control without lectin)), lectin-, saliva -And mutans + (mutans) only, lectin-, saliva + and mutans-(saliva only), lectin-, saliva-and mutans-(well only) Met. The suppression index of each condition was 100 for saliva + mutans, 20 for mutans alone, and no significant suppression index was shown for saliva alone and wells alone. The inhibition index of the mill extract is 90 when the protein concentration is 0.03125 mg / mL, 80 when the protein concentration is 0.0625 mg / mL, 60 when both the 0.125 mg / mL and 0.25 mg / mL. It was 55 when 0.5 mg / mL. The inhibition index of the willow matsutake extract is 145 when the protein concentration is 0.3125 mg / mL, 135 when the protein concentration is 0.625 mg / mL, 130 when the protein concentration is 1.25 mg / mL, and 2.5 mg / mL. At 100 and 65 at 5 mg / mL. From the above results, the effect of suppressing the adhesion and growth of S. mutans was observed in the range of 0.03125 mg / mL to 2 mg / mL in the mill extract, and only 5 mg / mL in the willow matsutake extract. . Therefore, the mill extract has an effect of suppressing the adhesion and growth of mutans bacteria at a concentration less than 1/100 of the willow matsutake extract, and the activity of the factor that suppresses the adhesion and growth of mutans bacteria is very high. It was shown to be expensive.

Purification of mill and willow matsutake lectins Mill and willow matsutake extracts were prepared as in Example 1. Each of the extracts was applied to a column packed with a carrier (GE Health Science, NHS-activated Sepharose) on which a glycoprotein (Asialofetuine, A4781, Sigma-Aldrich) was immobilized. After the substance that does not specifically bind to asialofetin was eluted with PBS, the substance that competitively inhibits the binding with asialofetwin by GalNAc was eluted with PBS containing 200 mM GalNAc. The protein concentration of the extract and the protein concentration of the purified lectin were measured using a BCA protein assay kit (Thermo Fisher Scientific Co., Ltd.).

Results of Purification Table 1 shows the amount and yield of lectin protein purified from mill and willow matsutake. 5.8 mg of sugar chain binding protein was collected from the mill, and 0.2 mg of sugar chain binding protein was collected from 49 g of willow matsutake. The lectin purification yields of mill and willow matsutake were 8.11% and 0.02%, respectively.

Biofilm Inhibition Assay with Purified Lectin The inhibitory effect of purified lectin on adhesion and growth of S. mutans was evaluated by the same procedure as in Example 1. As controls, ABA (Agaricus bisporus agglutinin (blood agglutinin from mushrooms), 300092, biochemical biobusiness) and BSA (bovine serum albumin, 81-003-3, Millipore) were used.

Results of Assay FIG. 2 is a graph showing the inhibitory effect of purified lectins derived from mill and willow matsutake on adhesion and growth of mutans bacteria. The error bar for each experimental condition indicates the standard deviation of the measured value of the experimental result repeated twice for each identical condition.

  The four bar graphs from the right end of FIG. 2 are the results of the control experiment. In order from the right end, lectin-, saliva- and mutans bacteria- (well only), lectin-, saliva- and mutans bacteria + (mutans bacteria). Only), lectin-, saliva + and mutans bacteria- (saliva only), and lectin-, saliva + and mutans bacteria + (saliva + mutans bacteria (positive control without lectin)). Met. The suppression index of each condition was 100 for saliva + mutans, 14 for mutans alone, and no significant suppression index was shown for saliva alone and wells alone. In addition, the inhibition indexes when the concentrations of ABA and BSA were 100 μg / mL were 24 and 97, respectively. The inhibitory indices for purified lectin concentrations of 0.5 μg / mL, 5 μg / mL and 40 μg / mL were 102, 76 and 26 for the mill and 91, 84 and 39 for the willow matsutake. From the above results, it was shown that the purified lectin of mill and willow matsutake remarkably suppresses adhesion and growth of mutans bacteria. In addition, 40 μg / mL mill purified lectin was shown to inhibit the attachment and growth of S. mutans to the same extent as 100 μg / mL ABA.

Inhibitory Effect of Mill-derived Protein Prepared by Another Protocol on Adherence and Growth of S. mutans. Preparation of Mill-derived Protein 17.49 kg of raw mill was dried at 40 ° C. The dried mill alga body having a dry weight of 1.5 kg was pulverized by mixer pulverization, 30 L of purified water was added, and the mixture was stirred at 15 ° C. for 16 hours to extract soluble components. From the solution containing the soluble component, solids were removed by continuous centrifugation at 4,000 × g, and the supernatant was collected. The supernatant was dried under reduced pressure at 60 ° C. The protein concentration of the extract was measured using a BCA protein assay kit (Thermo Fisher Scientific Co., Ltd.). A two-fold dilution series of the extract with PBS was prepared and subjected to an experiment to evaluate the effect on the adhesion and growth of mutans bacteria.

Preparation of protein from mushrooms and willow matsutake 9.1 g mushroom and 50 kg willow matsutake were dried and the soluble components were extracted with purified water after drying as in the mill described in the previous section. The protein concentration of the extract was also measured in the same manner as the mill.

Biofilm Inhibition Assay The inhibitory effect of the extract such as mill on the adhesion and growth of S. mutans was evaluated by the same procedure as in Example 1.

Results FIG. 3 is a graph showing the relationship between the protein concentration of the mill-derived purified water extract and the inhibitory effect on adhesion and growth of mutans bacteria. As is apparent from FIG. 3, the purified water extract derived from the mill had a protein index concentration rapidly decreased between 25 μg / mL and 50 μg / mL, and the 50% inhibitory concentration was 40 μg / mL.

  Table 2 below is a table showing the purification results of purified water extracts of mills, mushrooms and willow matsutake, and 50% inhibitory concentrations. The solids recovered from the purified water extract of mill, mushroom and willow matsutake were 461 g, 25.5 mg and 1550 g, respectively, and the protein content therein was 8.3 g, 9.9 mg and 506 g, respectively. Met. The 50% inhibitory concentrations of the purified water extract of mill, mushroom, and willow matsutake were 40 μg / mL, 90 μg / mL, and 4000 μg / mL, respectively.

  From the above results, it was revealed that in the purified water extract of this example, the mill can suppress the biofilm formation of mutans bacteria at the lowest concentration.

  The mill is almost tasteless and odorless seaweed, and there is a record that has been eaten in Japan as a sea pine since ancient times. Therefore, the mill extract or the purified lectin is not problematic from the viewpoint of safety. In addition, it is disclosed in Japanese Patent Application Laid-Open No. 9-48715 that an extract derived from seaweeds of a large number of green algae including the genus Mill exhibits antibacterial activity against the periodontal pathogen Porphyromonas gingivalis. It is disclosed. However, the publication discloses that the extract does not affect mutans bacteria, and that the antibacterial properties are not changed by either an extract by water or an extract by chloroform-methanol. In the extraction with chloroform-methanol, the protein is considered not to be distributed to the chloroform-methanol liquid layer due to denaturation, so the antibacterial component disclosed in the above publication cannot be a protein. Therefore, it is concluded that the caries preventive agent of the present invention is neither disclosed nor suggested in the publication.

Preparation of toothpaste A toothpaste is prepared according to the following formulation.
Calcium carbonate 50.00%
Glycerin 20.00%
Carrageenan 0.50%
Carboxymethylcellulose 1.00%
Lauryl diethanolamide 1.00%
Sucrose monolaurate 2.00%
Fragrance 1.00%
Saccharin 0.10%
Mill extract 0.05%
Water residue

Preparation of toothpaste A toothpaste is prepared according to the following formulation.
Aluminum hydroxide 45.00%
Gelling silica 2.00%
Sorbit 25.00%
Carboxymethylcellulose 1.00%
Sodium lauryl sulfate 2.00%
Fragrance 1.00%
Saccharin sodium 0.10%
Mill extract 0.04%
Water residue

Preparation of mouthwash A mouthwash is prepared according to the following formulation.
Ethanol 20.00%
Fragrance 1.00%
Polyoxyethylene hydrogenated castor oil 0.30%
Sodium monofluorophosphate 0.10%
Saccharin sodium 0.05%
Mill extract 0.04%
Water residue

Preparation of chewing gum Chewing gum is prepared according to the following recipe.
Sugar 53.00%
Gum base 20.00%
Glucose 10.00%
Fragrance 0.50%
Mill extract 0.10%
Minamata residue

Claims (8)

Characterized in that it comprises an aqueous solution extract from milled algae of mil family algae, anticaries agents.   The caries preventive agent according to claim 1, wherein the algae belongs to the genus Mill.   The algae belong to at least one species of the genus Mir selected from the group consisting of mill, isemomil, tamamil, cobsimil, chromyl, mottle mill, high mill, namban high mill, nezaci mill, hige mill, sakibutomiru, hiramil, and nagamil. The caries preventive agent according to claim 2, wherein   The caries preventive agent according to claim 3, wherein the algae is a mill. The dental caries preventive agent according to any one of claims 1 to 4, wherein the aqueous solution extract is a lectin that binds to a sugar chain that competes with GalNAc.   A pharmaceutical composition comprising the caries preventive agent according to any one of claims 1 to 5.   A dentifrice comprising the caries preventive agent according to any one of claims 1 to 5. A method for producing a caries preventive agent, comprising milling algae and extracting with an aqueous solution and formulating the obtained extract.

Images