WO2012043743A1 - Anti-bacterial composition and use thereof - Google Patents

Abstract

The present invention provides an anti-bacterial composition containing rice malt or an extract of the rice malt, as a novel anti-bacterial technique. This composition may additionally contain a fermentation raw material for producing the rice malt or an extract of the fermentation raw material.

Description

Antibacterial composition and use thereof

The present invention relates to an antibacterial composition and use thereof.

There are over 700 billion bacteria in the human oral cavity, and these bacteria interact to form dental plaque (dental plaque) on the tooth surface and gingival crevice. . Plaque increases when oral cleaning is inadequate. Gingivitis is an inflammation of the gums caused by plaque. Plaque reacts with calcium components in saliva to form tartar on pumice. Tartar is very hard and cannot be removed by normal brushing. Periodontal disease bacteria attached to the gingival sulcus penetrate into the periodontal pocket while melting the periodontal ligament and alveolar bone. Teeth whose gingiva is unable to maintain adhesion to the root will eventually fall out.

Periodontal disease is a disease in which periodontal tissues such as gums and alveolar bone are destroyed by periodontopathic bacteria in the oral cavity, and about 80% of adults are affected. Periodontal disease is roughly divided into gingivitis in which gingiva is inflamed and gingivitis in a stage that progresses deeply until alveolar bone resorption and periodontal ligament destruction occur. Among the periodontal diseases, a state where inflammation limited to the gingiva occurs is called gingivitis, and a state where inflammation occurs in other periodontal tissues is called periodontitis.

Periodontal disease was originally included in the category of adult diseases, but has recently been recognized as a lifestyle-related disease. It has also been found that periodontal disease increases the risk of developing arteriosclerosis and myocardial infarction. Furthermore, it has become clear that the symptoms of diabetes worsen with periodontal disease. In other words, it can be said that periodontal disease and diabetes have an adverse effect on each other. Indeed, evidence has shown that the bacteria responsible for periodontal disease are involved in systemic disease. That is, when periodontal disease bacteria settle in the periodontal pocket, the periodontal tissue can be destroyed and finally enter the blood. As a result, when periodontal disease bacteria are present in the blood, arteriosclerosis is likely to develop. In addition, “toxins” present in the cells of periodontal disease bacteria also adversely affect blood glucose levels. That is, endotoxin present in blood enhances the production of TNF-α from adipose tissue and liver. Since TNF-α also has an action of suppressing the intake of sugar in the blood, it inhibits the action of a hormone (insulin) that lowers the blood sugar level and makes it difficult to lower the blood sugar level. That is, in both periodontal disease and obesity, the secretion of TNF-α is activated, so that the blood glucose level cannot be controlled well, resulting in the onset of diabetes. Treatment of periodontal disease with antibacterial agents in diabetic patients with periodontal disease not only reduced the TNF-α concentration in the blood but also improved the HbA1c level, which indicates a controlled state of blood glucose level The result is obtained. As a result of caries bacteria and periodontal disease bacteria, it has already been clarified that a person with a heart prosthesis tends to develop endocarditis when undergoing treatment such as tooth extraction. This is because the blood flow is difficult to flow at the site of the valve, so that bacteria entering the body are easily infected and cause inflammation. In older people, the reflexes of swallowing and food are not performed well, which can cause oral bacteria to accidentally enter the respiratory tract and cause false pneumonia. Thus, the prevention and improvement of periodontal disease is important in preventing aspiration pneumonia.

The main causative agent of periodontal disease is Porphyromonas gingivalis (Pg), which is also a source of strong bad breath. Other than Pg, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Prevotella intermedia, etc. are said to be involved in periodontal disease. Pg is an anaerobic gram-negative bacilli and forms a black and malodorous colony on a blood plate. There is a capsule on the surface of Pg cells, which shows resistance to phagocytosis of phagocytes. Furthermore, the catalytic action of the protease produced by Pg not only damages the tissue but also degrades immunoglobulins such as IgG and IgA. Lipopolysaccharide present in the surface layer of Pg cells has the ability to promote bone resorption and adhere to mucosal epithelial cells and erythrocytes by cilia. Blood coagulation is also shown by the blood clotting factor (hemagglutinin) bound to Pg cells. Thus, Pg exhibits strong pathogenicity among oral bacteria and is frequently separated from the affected area of periodontitis occurring in adults.

Pg has 5 types of gene fimA encoding its own pilus protein. fimA is classified into types I to V based on the difference in the amino acid sequence of the encoded protein. Pili are involved in cell attachment and lead cells to apoptosis. Toxicity due to Pg is determined by this ability to induce apoptosis, and the toxicity is strong in the order of type II, type IV, and type III. The most toxic type II is frequently detected from patients with severe periodontal disease, and the least toxic type V is also detected from healthy subjects.

Techniques aimed at treating or preventing periodontal disease using microorganisms have been disclosed (see Patent Documents 1 to 5). For example, Patent Document 1 discloses that lactic acid bacteria, lactic acid bacteria culture solutions, and the like have an effect of suppressing the growth of oral pathogenic bacteria (for example, Pg and mutans bacteria that cause dental caries) and sweeteners such as xylitol. It is described that saccharides such as food and sugar, and various plant extracts, alone, do not exhibit the effect of inhibiting the growth of oral pathogenic bacteria, but significantly enhance the effects of the lactic acid bacteria, lactic acid bacteria culture medium, and the like. Patent Documents 2 to 4 disclose lactic acid bacteria that inhibit the growth of Pg, and Patent Document 4 further enhances the effect by further containing a saccharide that can be assimilated by the lactic acid bacteria described in the literature, It is described that lactic acid bacteria described in the literature significantly suppress the secretion of enzymes from Pg. Patent Document 5 discloses a preventive agent for caries or periodontal disease, which contains live yeast or a mixture of live yeast and dead yeast, and improves immune resistance in periodontal tissue. In order to further increase, it is described that a killed edible bacterium other than yeast is further added.

The toxin produced by Pg is a kind of protease and is called gingipain. This enzyme damages host cells by degrading in vivo proteins, causing various pathological conditions related to periodontal disease, and by destroying vascular tissue, Pg enters the blood vessels, causing arteriosclerosis and It has been found that it is also the cause of heart disease. Cysteine protease inhibitors such as antipain and leupeptin are known as substances that inhibit gingipain, but are not used for the prevention and / or treatment of periodontal disease because of their high toxicity. Patent Documents 6 and 7 disclose a gingipain inhibitor using a plant or a plant extract as an active ingredient in consideration of prevention and / or treatment of periodontal disease.

International Publication Gazette “WO2002 / 080946 Pamphlet (International Publication on Oct. 17, 2002)” Japanese Patent Publication “Japanese Patent Laid-Open No. 2010-124772 (published on June 10, 2010)” Japanese Patent Publication “Japanese Patent Laid-Open No. 2006-328052 (published on December 7, 2006)” Japanese Patent Publication “JP 2003-171292 (June 17, 2003)” Japanese Patent Publication “Japanese Patent Laid-Open No. 2006-219441” (published on August 24, 2006) Japanese Patent Publication “JP 2003-335648 A (published on November 25, 2003)” Japanese Patent Publication “Japanese Patent Laid-Open No. 2007-91703 (published on April 12, 2007)”

Thus, for the purpose of prevention and / or treatment of periodontal disease, research on substances that achieve Pg growth inhibition and gingipaine activity inhibition is underway. However, most reports on lactic acid bacteria have been reported on Pg growth inhibition using microorganisms, and most reports on specific plant extracts have been reported on gingipaine activity-suppressing components.

The present invention has been made in view of the above-described problems, and an object thereof is to provide a new technique relating to antibacterial, particularly a new technique for treating periodontal disease.

In order to solve the above-mentioned problems, the composition of the present invention is an antibacterial composition, and is characterized by containing koji or an extract of the koji. It is preferable that the composition of the present invention further contains a fermentation raw material for obtaining the koji or an extract of the fermentation raw material.

In the composition of the present invention, the fermentation raw material for obtaining the koji is preferably selected from the group consisting of soybeans, defatted soybeans, peanuts, empty beans, wheat, rice, and soy sauce koji. Further, in the composition of the present invention, the koji is used for brewing from Aspergillus filamentous fungi A.zaoryzae, A. sojae, A. kawachii and A. awamori used to ensure (secure) food safety. It is preferable to be produced using a koji mold selected from the group consisting of More preferably, the fermentation raw material for obtaining the koji is selected from the group consisting of soybeans, defatted soybeans and soy sauce koji materials, and the koji is A. oryzae.

The composition of the present invention is preferably used for application in the oral cavity, whether used to treat periodontal disease, used to inhibit Candida growth, May be used to inhibit the growth of H. pylori. That is, by using the composition of the present invention, it is possible to treat periodontal disease, suppress the growth of Candida in the oral cavity, and further suppress the growth of Helicobacter pylori in the stomach. Can do. Thus, since the active ingredient in the composition of the present invention has a broad antibacterial spectrum, it can be said that the composition of the present invention is an antibacterial composition.

In addition, the composition of the present invention may be used as an external preparation from the viewpoint of use for suppressing the growth of Candida, and is preferably used as a cosmetic composition.

The composition of the present invention may be a quasi-drug, a food, or a medicine, and when used in these, the extract of koji is preferably a water extract. The fermentation raw material extract is also preferably a water extract.

The method of the present invention is characterized by including the step of bringing the composition into contact with a causative agent of periodontal disease in the oral cavity of the subject in order to treat periodontal disease. The method of the present invention may further include the step of supplying the composition into the oral cavity of the subject.

The method of the present invention is also characterized by including the step of contacting the above composition with Candida in the oral cavity of a subject in order to treat oral candidiasis. The method of the present invention may further include the step of supplying the composition into the oral cavity of the subject.

The method of the present invention is also characterized by including the step of bringing the composition into contact with H. pylori in the stomach of a subject in order to inhibit the growth of H. pylori. The method of the present invention may further comprise the step of supplying the composition into the stomach of a subject.

If the present invention is used, the growth of periodontal disease bacteria can be inhibited. Moreover, if this invention is used, the toxin which a periodontal disease microbe produces | generates can be inhibited. By using these in combination, the action of periodontal disease bacteria can be more significantly suppressed. Furthermore, since Pg which is a causative agent of periodontal disease is also a main cause of bad breath, if this invention is used, bad breath can be prevented or improved.

If the present invention is used, periodontal disease can be treated, the growth of Candida in the oral cavity can be suppressed, and further, the growth of Helicobacter pylori in the stomach can be suppressed. Thus, the present invention provides an antimicrobial composition.

It is a figure which shows the result of having investigated the growth inhibitory activity with respect to Pg using various koji molds and various koji materials. It is a figure which shows the anti- gingipain activity by a soybean cake and its extract, and a soy sauce cake raw material and its extract. It is a figure which shows the anti-Candida activity by the extract of various cocoons. It is a figure which shows the anti- H. pylori activity by the extract of various cocoons. It is a figure which shows anti-periodontal disease microbe activity by the active ingredient of this invention. It is a figure which shows the minimum growth inhibitory concentration of an anti-periodontal disease microbe by the active ingredient of this invention. It is a figure which shows anti-periodontal disease fungal activity by the composition for oral cavity containing the active ingredient of this invention. It is a figure which shows the anti- H. pylori activity by the active ingredient of this invention.

Hereinafter, the embodiment of the present invention will be described taking treatment of periodontal disease as an example. However, the present invention is not limited to this, and can be applied not only to periodontal disease bacteria but also to H. pylori and Candida bacteria. Antibacterial composition and use thereof.

[1] Active ingredient for treating periodontal disease [Koji]
Aspergillus is essential for brewing traditional Japanese fermented foods such as soy sauce, miso, sake and shochu. Neisseria gonorrhoeae is a general term for molds used in making strawberries.

As used herein, “gonococci” refers to Aspergillus fungi that have been secured for safety based on long-term eating experience. In general, there exists a mold belonging to the genus Monascus called Monascus (such as Monascus purpureus), but the Monascus genus is completely different from the Aspergillus genus in form. Aspergillus fungi usually have a colorless mycelium that stretches and branches. Aspergillus fungi are shaped like a bulbous vesicle at the apex of the conidophore that emerges from the hyphae during asexual reproduction, and the top of the top sac is radiated on top. A phialide is formed, and conidia grows in a chain at its tip. Neisseria gonorrhoeae is sometimes referred to as black koji mold or yellow koji mold depending on the color of its conidia. By the way, mold (Monasucus purpureus) used to make red sake in China, etc., produces a red or pink pigment in the mycelium, and at the time of sexual reproduction, the ascomyce that contains a large number of ascombs is used as the tip of the mycelium. The ascending cones that have formed and disappeared during asexual reproduction are present as single ascending.

Patent Document 1 is a document in which lactic acid bacteria, lactic acid bacteria culture solutions, and the like show the growth inhibitory effect of oral pathogens, and it is described that the koji mold remarkably enhances the growth inhibitory effect of Pg by lactic acid bacteria culture solutions, similar to xylitol. . However, according to Table 18 of Patent Document 1, it can be easily understood that there is no difference in the effect of suppressing the growth of Pg between the case where the koji mold is added and the case where the koji mold is not added. Of course, according to Table 18 of Patent Document 1, it cannot be understood that the koji mold itself has a Pg growth inhibitory effect. Thus, a person skilled in the art who understands that the Monascus genus Koji mold is completely different from the Aspergillus genus Koji mold, and that the Koji mold is not involved in Pg growth inhibition, The invention cannot be easily conceived.

Aspergillus gonococcus produces enzymes such as amylase, protease, and peptidase, which degrades starch and protein as raw materials and converts them into glucose and amino acids. These degradation products contribute to the flavoring characteristic of brewing. Most of the koji making is done using Aspergillus oryzae, and A. oryzae is also used in the production of sake and miso. In the production of soy sauce, 90% of it is A. oryzae and the remaining 10% is A. sojae. As other koji molds, A. kawachii used for shochu making in Kyushu and A. awamori used for producing Awamori in Okinawa are also known.

As shown in the examples described later, if a fermented product of koji mold or an extract thereof is used, the growth of periodontal disease bacteria can be inhibited. Can inhibit. If the koji raw material and the koji produced from the koji raw material are used together, Pg growth inhibition and toxin inactivation can be performed simultaneously, so that the action of periodontal disease bacteria can be more significantly suppressed. In addition, the extract of defatted soybean meal produced from defatted soybeans sterilized at 120 ° C for 30 minutes as a raw material not only significantly inhibits the growth activity of periodontal disease bacteria, but also causes Helicobacter pylori and candidiasis It also significantly inhibits the growth of fungi (Candidasarbicans).

In the present specification, “salmon” is also referred to as a fermentation product or a fermentation product by koji mold, and “salmon raw material” is intended to be a raw material used in the production of persimmons to obtain a fermentation raw material or persimmon. It is also called a fermentation raw material.

[Active substances against periodontal disease bacteria]
As shown in the Examples described later, cocoons or extracts thereof exhibit growth inhibitory activity against periodontal disease bacteria. That is, cocoon or an extract thereof has an activity of inhibiting the growth of periodontal disease bacteria. Furthermore, sputum or its extract has the activity of inhibiting the growth of candidiasis causative bacteria (Candida arbicans) and gastric ulcers and Helicobacter pylori, which is a risk factor for gastric cancer.

The koji mold used for producing the target koji is not particularly limited as long as it does not produce a toxin such as aflatoxin, but is selected from the group consisting of A. oryzae, A. sojae, A. kawachii and A. awamori. The selected gonococci will favorably inhibit Pg growth, with A. oryzae being particularly preferred. Moreover, the raw material for straw used for producing the target straw is not particularly limited as long as it is a grain that can be used for fermentation by Aspergillus spp., But soybean, wheat and rice are preferred, and soybean and wheat are particularly preferred. As the koji material, defatted soybean and soy sauce koji materials (that is, a mixture of defatted soybean and wheat) are also preferably used. The target koji is not particularly limited as long as it is a koji obtained by Aspergillus spp., But soy sauce koji, soybean koji, defatted soybean koji, wheat koji and rice koji are preferred, soy sauce koji, soybean koji, defatted soybean koji and Wheat straw is more preferred, soy sauce lees, soybean meal and defatted soybean meal are particularly preferred.

The following procedures are used to produce soy sauce: (1) Inoculation with koji molds in raw materials (mixture of soy materials boiled and steamed (defatted soybeans) and roasted and broken wheat materials) (2) The obtained koji is poured into saline solution in a charging tank and fermented and matured to obtain soy sauce moromi; (3) The soy sauce as a product is squeezed, fired, etc. obtain. Since said (1) is normally performed over several days, it can be said that the starch and protein which are contained in the raw material are not contained in the manufactured koji. That is, as used herein, “koji” is a raw material grain (eg, rice, soybeans, wheat, etc.) inoculated with koji mold and propagated, and the starch and protein of the raw material are completely degraded. A state is intended.

The solvent used for the extraction of koji is not particularly limited, but for example, water (including hot water), alcohol (for example, methanol, ethanol, propanol, etc.), hydrochloric acid, organic acid (for example, citric acid, acetic acid, etc.) ) And organic solvents (chloroform, ethyl acetate, etc.). From the viewpoint of use in foods, water is most preferable. If necessary, stirring may be performed using an ultrasonic wave, a stirrer, or the like during extraction.

[Active substances against toxins produced by periodontal disease bacteria]
As shown in the examples described later, the raw material for salmon or its extract exhibits inhibitory activity against gingipain, which exhibits trypsin-like activity. That is, the raw material for salmon or its extract inhibits the activity of gingipain. Gindipain is a cysteine protease produced by Pg. There are two enzymes, Arg-gingipain (Rgp) that cleaves at the position of arginine with different peptide cleavage site specificities and Lys-gingipain (Kgp) that cleaves at the position of lysine. .

Gindipain, a proteolytic enzyme secreted by Pg, is a pathogenic factor itself and acts as a toxin, but is also an important factor for Pg growth. As shown in Examples described later, soybean meal or an extract thereof inhibits Pg growth and gingipain activity. On the other hand, the soy sauce koji raw material or its extract does not inhibit the growth of Pg, but strongly inhibits the activity of gingipain. From these facts, it is considered that the inhibition of gingipaine activity by the koji raw material or its extract acts on a pathway independent of the growth of periodontal disease bacteria.

The raw material of the koji used to produce the target koji is not particularly limited as long as it is a cereal or legume, but when used together with the koji mentioned above, it is used for fermentation by Aspergillus spp. Preferred grains are preferred, soy (including defatted soy), wheat and rice are more preferred, soy (including defatted soy) and wheat are particularly preferred. Moreover, as mentioned above, defatted soybeans and soy sauce koji materials (that is, a mixture of defatted soybeans and wheat) are also preferably used as koji materials.

The solvent used for the extraction of the raw material is not particularly limited. For example, water (including hot water), alcohol (eg, methanol, ethanol, propanol, etc.), hydrochloric acid, organic acid (eg, citric acid, acetic acid) And organic solvents (chloroform, ethyl acetate, acetone, hexane, etc.). From the viewpoint of use in foods, water is most preferable. If necessary, the extraction may be performed using ultrasonic waves, a stirrer, an ultrahigh pressure (100 mPa) treatment corresponding to a depth of 10,000 meters at the seabed, or the like.

[2] Utilization of active ingredient (1) Composition In one aspect, the present invention provides a composition containing an active ingredient for treating periodontal disease. As used herein, the term “treatment” is intended to alleviate, ameliorate, or eliminate symptoms, and can be performed therapeutically (after onset) as well as prophylactically (before onset). What can be done is also encompassed. The “active ingredient for treating periodontal disease” is intended to be at least one of the above-mentioned “active substance against periodontal disease bacteria” and “active substance against toxin produced by periodontal disease bacteria”. It may be abbreviated as “active ingredient”. That is, as used herein, the “active ingredient” can be koji or an extract thereof, and can be a koji raw material or an extract thereof. Since periodontal disease bacteria are also a cause of bad breath, as used herein, “treatment of periodontal disease” can include “preventing or improving bad breath”.

As used herein, the “active ingredient” is “added 3 times the weight of water, then extracted for 1 hour with shaking at room temperature, and filtered (0.2 μm It is quantified on the basis of “a dry powder obtained by freeze-drying a sterilized extraction solution”. Specifically, as shown in the examples described later, 60 mL of water was added to 20 g of cocoon for extraction, and 30 ml of extraction solution was freeze-dried to obtain a suisui extract (3.1 g). The amount of active ingredient is defined in the standard. Moreover, in this specification, the extraction solution mentioned above is used as an antibacterial activity sample of a koji extract.

The composition of the present invention may be a solid agent dissolved in water before use, a liquid such as a suspension, or a paste preferable for application. The composition of the present invention contains an active ingredient, and the shape is not particularly limited as long as it contains an effective amount for expressing its physiological action, and is in the form of tablets, granules, capsules Orally ingestible forms such as are also included. In addition, the content of the active ingredient in the composition of the present invention is not particularly limited, and can be appropriately selected from the viewpoint of its functionality and expression of action.

The composition of the present invention can take any form, but is intended to prevent or treat periodontal disease, or prevent or improve bad breath, and is preferably in a form that easily stays in the oral cavity. , Powder, granule, paste, pill, chewable, gargle, troche, patch, etc., tablet, powder, granule, paste, chewable, gargle, troche, or patch It is more preferable that

Since it aims at prevention or treatment of periodontal disease or prevention or improvement of bad breath, the composition of the present invention is preferably provided as an oral composition. The “oral composition” is not limited as long as it is a form applicable to the oral cavity, and may be a quasi-drug, an edible composition, or a pharmaceutical composition. The active ingredient of the present invention can be blended in various known oral compositions, and the active ingredient of the present invention is blended in the range of 0.001 to 80% by mass with respect to the total amount of the composition. More preferably, it can be blended within the range of 0.01 to 50% by mass, more preferably 0.1 to 50% by mass, and still more preferably 1 to 50% by mass, and is suitably used for the practice of the present invention. From about 30 g of the antibacterial activity sample obtained, about 3 g of the active ingredient of the present invention can be obtained, and from the relationship between the weight of the active ingredient of the present invention described in the Examples and the minimum inhibitory concentration against Pg, the active ingredient of the present invention It is still more preferable that the content is 2.5 to 25% by mass with respect to the total amount of the composition.

In addition, the composition for oral cavity can be prepared according to a conventionally well-known method, and may be prepared by combining any component that is usually blended alone or in combination according to the dosage form. Examples of the components to be blended include abrasives, surfactants, binders, thickeners, sweeteners, preservatives, fragrances, colorants, pH adjusters, excipients, various medicinal ingredients, and the like. However, it is not limited to these.

Preferred oral compositions for the present invention include dentifrices (toothpastes, powder dentifrices, liquid dentifrices), chewable tablets, mouthwashes, mouthwashes (including tablets and powders), ointments, creams (gingival massage). Cream), mouth fresheners, denture cleaners, troches, gums and the like, but are not limited to these.

Favorable edible compositions for the present invention include, but are not limited to, candy, chocolate, drinks, fermented products and the like. When fermented products are used as edible compositions, various ingredients known to be incorporated in fermented foods or fermented beverages, such as sweeteners, flavoring agents, thickeners, flavoring agents, coloring agents, wearing agents, etc. A fragrance | flavor etc. may be mix | blended. The edible composition of the present invention is an edible composition for inhibiting the growth of periodontal disease bacteria or toxins from periodontal disease bacteria, and is extremely useful as a health food (functional food). The production method of the edible composition of the present invention is not particularly limited, and examples thereof include cooking, processing, and production by a generally used method for producing foods or beverages. The active ingredient described above is contained in the food or beverage. It should just be contained.

Preferred pharmaceutical compositions for the present invention include, but are not limited to, sustained release formulations for administration into periodontal pockets. When provided as a pharmaceutical composition, the composition of the present invention can take a desired dosage form using a known method in the pharmaceutical field. For example, after forming as a solid agent by adding excipients, binders, disintegrating agents, lubricants, coloring agents, flavoring agents, etc., if necessary, powders, fine granules, granules , Tablets, coated tablets, capsules and the like.

The pharmaceutical carrier used in the pharmaceutical composition of the present invention can be selected according to the administration form and dosage form of the pharmaceutical composition. In the case of oral preparations, for example, starch, lactose, sucrose, mannitol, carboxymethylcellulose, corn starch, inorganic salts and the like are used as pharmaceutical carriers. Moreover, when preparing an oral preparation, you may mix | blend a binder, a disintegrating agent, surfactant, a lubricant, a fluidity promoter, a corrigent, a coloring agent, a fragrance | flavor, etc. further.

The pharmaceutical composition of the present invention can be produced by a known method in the pharmaceutical field. The content of the active ingredient in the pharmaceutical composition of the present invention is not particularly limited as long as it is an amount that allows the pharmaceutical composition to be administered in an appropriate amount in consideration of the administration form, administration method and the like. The dosage of the pharmaceutical composition of the present invention is appropriately set according to the formulation form, administration method, purpose of use, and age, weight, and symptom of the patient to whom the pharmaceutical is administered, and is not constant. Administration may be carried out in a single dose or divided into several doses within a desired dose range. Moreover, the pharmaceutical composition of the present invention can be orally administered as it is, or can be added to any food or drink and taken daily.

As used herein, the term “composition” may be an embodiment in which the component is contained in a single composition or an embodiment in which the component is separately provided in a single kit. . That is, as used herein, “kit” can be said to be an embodiment of “composition”.

The above-mentioned quasi-drug can be used as a component of an external preparation (for example, a cosmetic composition) applied not only to the oral cavity but also to the skin as a natural preservative that suppresses the growth of Candida. That is, in another aspect, the present invention provides a composition containing an active ingredient for inhibiting the growth of Candida. The composition of the present invention is a quasi-drug used as an external preparation (for example, a cosmetic composition). The composition according to the present invention contains an active ingredient for suppressing the growth of Candida. As an external preparation, various usage forms such as a medicinal skin external preparation, a cosmetic skin external preparation, and a cosmetic composition can be used. As used herein, the term “skin” is intended for skin of the face, neck, chest, back, arms, legs, hands and scalp.

The external preparation as a cosmetic composition may be present in a form usually used in cosmetics. The cosmetic composition according to the present embodiment can be prepared in various forms according to a conventional method, and even if it is newly produced by containing an active ingredient for suppressing the growth of Candida, It may be produced by adding the above-mentioned active ingredient to a conventional cosmetic. The cosmetic composition according to the present embodiment comprises an oil, a moisturizer, an ultraviolet absorber, a whitening agent, an alcohol, a chelating agent, a pH adjuster, an antiseptic, a thickener, and a pigment that are generally used as cosmetic ingredients. , Plant extracts, fragrances and the like can be combined in any combination. Cosmetic compositions include various uses and forms such as water / oil or oil / water type emulsified cosmetics, creams, cosmetic emulsions, lotions, oily cosmetics, lipsticks, foundations, skin cleansers, hair artic , Hair claims, hair styling agents, hair nourishing agents, hair restoration agents, bathing agents.

The cosmetic composition according to this embodiment may be a fluid or a solid. When provided in a fluid, it may have the appearance of a white or colored cream, ointment, emulsion, lotion, serum, paste, or mousse. Furthermore, the cosmetic composition according to this embodiment may be applied in the form of an aerosol. The cosmetic composition according to this embodiment can also be provided in a solid form, in particular in the form of a stick.

In using the cosmetic composition according to this embodiment, a small amount, for example, 1 to 100 mL of the composition is applied to the exposed surface of the skin from a suitable container or applicator and, if necessary, hand or finger or suitable Using an instrument, spread and rub into the skin.

In one embodiment, the composition according to the invention may be formulated as a lotion, cream or gel. The composition according to this embodiment can be packaged in a container suitable for its viscosity and intended use. The composition according to this embodiment, for example, the lotion or cream is packaged in a bottle or roll-ball applicator, or a propellant-driven aerosol device or a pumped container suitable for finger operation. obtain. When the composition according to this embodiment is a cream, it may be contained in a non-deformable bottle or squeeze container, for example, a wide-mouth bottle with a tube or a lid, or enclosed in a capsule. The composition according to this embodiment can be provided in a form housed in a closed container containing a cosmetically acceptable composition.

Examples of external preparations other than cosmetic compositions include solid, semisolid or liquid preparations for transdermal administration, suppositories, and the like. For example, emulsions such as emulsions and lotions, liquid preparations such as external tinctures, ointments such as oily ointments and hydrophilic ointments, patches for transdermal administration such as films, tapes, and poultices It can also be used.

Examples of medicinal skin external preparations and cosmetic skin external preparations include various ointments containing medicinal ingredients. The ointment may be one based on an oily base, or one based on an oil / water or water / oil type emulsion base. The oily base is not particularly limited, and examples thereof include vegetable oils, animal oils, synthetic oils, fatty acids, and natural or synthetic glycerides. The medicinal components are not particularly limited, and for example, analgesic / anti-inflammatory agents, analgesics, bactericides / disinfectants, astringents, emollients, hormones, vitamins and the like can be used as needed. .

The external preparation according to this embodiment can be produced by a known method in the pharmaceutical field. In the external preparation according to the present embodiment, the content of the active ingredient for suppressing the growth of Candida is Candida in a desired dosage range using the skin external solvent in consideration of the administration form, administration method, and the like. If it is the quantity which can administer the active ingredient for suppressing the proliferation of this, it will not specifically limit. Moreover, the compounding quantity of the active ingredient for suppressing the proliferation of Candida bacteria to the external preparation which concerns on this embodiment is not restrict | limited in particular.

(2) Kit In one aspect, the present invention also provides a kit comprising an active ingredient for treating periodontal disease. In another aspect, the present invention provides a kit comprising an active ingredient for suppressing the growth of Candida. As used herein, the term “kit” intends a package with a container (eg, bottle, plate, tube, dish, etc.) containing a particular material. Preferably, an instruction for using the material is provided. As used herein in the context of a kit, “comprising” is intended to mean being contained within any of the individual containers that make up the kit. Further, the kit of the present invention may be a package in which a plurality of different compositions are packed together, or may be a solution in a form of a solution contained in a container. The kit of the present invention may be prepared by mixing two or more different substances in the same container or in separate containers. The “instructions” may be written or printed on paper or other media, or may be affixed to electronic media such as magnetic tape, computer readable disk or tape, CD-ROM, etc. . The kit of the present invention may be used to constitute the above-described composition, and may comprise separately the substances contained in the above-described composition, or separately comprise the above-described composition and further components. May be.

(3) Method for treating periodontal disease The present invention further provides a method for treating periodontal disease. The method of the present invention includes a step of bringing the active ingredient of the present invention (or a composition containing the active ingredient) into contact with the causative bacteria of periodontal disease. The step of bringing the active ingredient of the present invention into contact with the periodontal disease causative agent is to include a troche or chewable agent in the mouth even if it is toothpaste, finger paste application, patch application. Alternatively, it may be a step of gargleing with a gargle. In this step, the time for contacting the active ingredient of the present invention with the causative agent of periodontal disease is appropriately designed according to the means employed, but is preferably in the range of 10 seconds to 10 minutes, 30 More preferably within the range of seconds to 10 minutes, even more preferably within the range of 1 minute to 5 minutes.

Since the method of the present invention requires that the active ingredient of the present invention and the causative bacteria of periodontal disease be brought into contact with each other, the administration to a subject is preferably oral administration. That is, the method of the present invention may further include a step of supplying an active ingredient for treating periodontal disease into the oral cavity of the subject.

Those skilled in the art who read this specification will easily understand that the specific application of the active ingredient in the method of the present invention may be in accordance with the use form of the composition and kit described above.

[3] Further use of the present invention As described above, the present invention can be applied not only to periodontal disease bacteria but also to H. pylori and Candida. That is, the present invention provides an active ingredient for inhibiting the growth of Helicobacter pylori and Candida, a composition containing the active ingredient, a kit comprising the active ingredient, and the active ingredient, the composition, and the Provided is a method for inhibiting the growth of Helicobacter pylori and Candida using a kit.

Candida is a fungus that inhabits the human oral cavity and the like, and is a causative bacterium of a disease (oral candidiasis) that mainly forms moss-like small spots. This bacterium is resident in about 30-40% of human oral cavity. Oral candidiasis occurs when the defense function against immune infection is reduced, or when an antibacterial agent having no effect on this bacterium is administered over a long period of time. Oral candidiasis is usually seen in the elderly and infants, but can be caused by diabetes, cancer radiotherapy or chemotherapy, long-term use of steroids or antibacterials, immunodeficiencies (such as AIDS), and the like. In addition, a decrease in the amount of saliva, keeping dentures, poor oral cleaning, etc. can also cause oral candidiasis.

Also, more than 70% of Japanese over 50 years old are infected with Helicobacter pylori. H. pylori is a microaerobic and spiral gram-negative bacterium. This bacterium adheres to the gastric mucosa in the stomach and survives, but can move while swimming in mucus by the rotational movement of polar flagella located at both ends of the long axis of the microbial cell. H. pylori has been found to be a risk factor for inflammatory diseases such as atrophic gastritis and gastric ulcers, and gastric cancer. For the eradication of Helicobacter pylori for the purpose of preventing and treating gastric ulcer and gastric cancer, three-drug combination therapy in which a proton pump inhibitor is added to two antibiotics such as amoxicillin and clarithromycin is common. However, as a side effect of this therapy, epithelial colitis and renal failure are likely to relapse, and in addition to abnormal taste and diarrhea, the appearance of resistant bacteria is often observed.

H. pylori produces an enzyme (urease) that decomposes urea into CO ₂ and ammonia. As a result, by neutralizing gastric acid with ammonia, H. pylori can survive in a gastric juice environment having a low pH. In addition, urease is greatly involved in the chemotaxis and gastric mucosal damage of Helicobacter pylori, and cell vacuolating toxin (VacA) and mucinase secreted by Helicobacter pylori also damage the gastric mucosa and gastric epithelial cells. On the other hand, when H. pylori injects CagA toxin into gastric mucosal cells, interleukin 8 (IL-8) is induced / produced from the gastric mucosal cells, and as a result, polynuclear leukocytes are assembled. These leukocytes produce active oxygen and attack H. pylori, but also damage gastric mucosal cells themselves, causing atrophic gastric ulcers and the development / progress of cancer. Therefore, there is an urgent need to develop a method for eradicating H. pylori that is effective against H. pylori and is less invasive.

JP-A No. 2001-143 discloses a food / beverage product for sterilization / infection prevention of Helicobacter pylori containing a specific lactic acid bacterium (Lactobacillus gasseri). Japanese Patent Application Laid-Open No. 9-241173 discloses a drug and fermented food that can sterilize H. pylori from the stomach or duodenum containing a specific lactic acid bacterium (Lactobacillus salivarius or Lactobacillus brevis) as an active ingredient. In addition, the present inventors have acquired new plant lactic acid bacteria and manufactured fermented beverages having new flavors and functions using these lactic acid bacteria (see Japanese Patent Application Laid-Open No. 2006-42796). However, these reports are all technologies using lactic acid bacteria.

The present inventors have recently found that anti-pylori activity exists in the culture supernatant when gonococci are cultured using citrus fruit juice, and that the main body of the anti-pylori activity is hesperetin, an aglycon of hesperidin. (Patent pending, currently unpublished). JP-A-2003-102430 describes that hesperidin contained in citrus is converted to hesperetin by fermenting citrus using black koji mold. In addition, in order to effectively use hesperetin that has low solubility in water and easily precipitates, a technique for stably dispersing hesperetin in an aqueous solution has been proposed (see Japanese Patent Application Laid-Open No. 2008-184385).

As shown in the examples described later, the extract of defatted soybean meal produced using defatted soybeans sterilized at 120 ° C. for 30 minutes as a raw material, not only significantly inhibits the growth activity of periodontal disease bacteria, Helicobacter pylori) and Candida arbicans are also significantly inhibited. The main body of anti-pylori activity in the present invention is not hesperetin.

In the present specification, the specific application of the active ingredient in the composition, kit and method of the present invention for inhibiting the growth of Helicobacter pylori and Candida may conform to the above-mentioned form of anti-periodontal disease. Those skilled in the art who read the book will readily understand.

It should be noted that all of the academic literatures and patent literatures described in this specification are incorporated herein by reference.

The present invention will be described in more detail by the following examples, but should not be limited thereto.

[1. Growth inhibition of Pg by sputum]
Using 4 types of Aspergillus (A. oryzae, A. sojae, A. kawachii, A. awamori), ferment rice, wheat, soy sauce raw material (a mixture of defatted soybean and wheat), soybeans, broad beans and peanuts A firewood was manufactured.

To manufacture soybean meal, first, the soybean is lightly washed and then immersed in water until the weight is about 1.6 times that of the soybean raw material. Since the water absorption rate during immersion varies depending on the temperature of the immersion water and the type of soybean, it is necessary to conduct a preliminary test each time. Specifically, after preparing 100 g of various round soybeans, each was washed, and each round soybean taken out after 30, 60, 90, 120 minutes immersed in water was wrapped in gauze, and lightly shaken to adhere to the surface. Remove moisture. Next, the weight of each type of whole soybean is measured and the approximate time to reach about 160 g is examined. If the amount of water absorption is too large, it becomes easy to be contaminated with bacteria other than Neisseria gonorrhoeae, and if it is too little, soybeans cannot be steamed successfully, and as a result, it is difficult to undergo enzymatic degradation of Neisseria gonorrhoeae. The supernatant after absorbing water is discarded, and the absorbed soybean is placed on a towel and drained for about 2 hours. The soybean is then wrapped in a mesh cloth and autoclaved at 121 ° C. for 30 minutes. The sterilized soybean is cooled to around 35 ° C. and then inoculated with Aspergillus oryzae conidia. After culturing at 30 ° C. for 24 hours, the temperature is lowered to 25 ° C. and the mixture is further cultured for about 18 hours. The iron making is performed in a place where drying is not possible and the temperature can be maintained. It is desirable to absorb the water droplets that fall due to the heat generated by the koji mold during filter making using filter paper or the like. When preparing a koji extract, use 2 to 3 times the koji weight of water at room temperature with shaking for 2 hours. Concentrate the obtained extract solution as appropriate to obtain a koji extract. obtain. Of course, when the amount of water is large, the concentration of the extract decreases. In addition, although the soybean was mentioned as an example as a fermentation raw material and the manufacture of the koji was demonstrated, the other fermentation raw materials may be performed according to this.

Then, the function of the extract from the obtained straw was examined. In particular, it was examined whether or not it exhibits antibacterial activity against Porphyromonas gingivalis (hereinafter referred to as Pg), which is a causative agent of periodontal disease and can also be a cause of bad breath. For Pg, HNA99 (type V) distributed from Pediatric Dentistry, Hiroshima University Dental School was used.

The extract of persimmon of the present invention is usually obtained using water, but as a solvent for extraction, in addition to water, an organic solvent such as ethanol, ethyl acetate, chloroform, acetone, butanol, hexane, or The mixed solvent which combined these in the appropriate ratio, especially the mixed solvent which combined the hydrophilic organic solvent with water are mentioned. Moreover, various fruit juices and vegetable juices can also be used as a solvent for extraction. From the viewpoint of food safety, the koji extract is preferably a water extract, and the fermentation raw material extract is also preferably a water extract. The extraction temperature may be between 0 ° C. and 100 ° C., but room temperature is the most desirable. The extraction time is in the range of 1 hour to 48 hours, preferably about 2 hours while shaking, and is preferably performed appropriately depending on the raw material of the straw and the straw.

A bioassay using Pg was performed according to the following procedure. All Pg cultures were performed at 37 ° C. under anaerobic conditions. Pg was cultured for 48 hours using PV Brucella HK agar medium (BHK PV, Far East). 28 g of brucella broth powder (manufactured by BD) was dissolved in 1 L of pure water to prepare 2.8% brucella broth. One platinum loop of Pg was collected and suspended in 140 μL of 2.8% Brucella broth. This suspension (130 μL) was applied on the surface of a plate containing BHK PV. This Pg-coated plate was used as an assay plate. A sterilized paper disc (8 mm in diameter) impregnated with 30 μL of sample (such as sputum extract) was then placed on the assay plate. Pg was cultured for 48 hours after placement and antibacterial activity was evaluated based on the size of the resulting inhibition circle diameter.

In soybean koji and wheat koji, even if any of Koji molds (A. awamori SH41, A. kawachii SH46, A. oryzae S-03, A. sojae No.9) is used, a blocking circle against Pg is formed (FIG. 1). In particular, the koji obtained using A. oryzae had very strong antibacterial activity against Pg, and a Pg inhibition circle was formed even in rice koji.

[2. Growth inhibition of Pg by straw or its raw material]
The antibacterial activity of soy sauce cake and soybean meal when A. oryzae was used was further investigated using soy sauce cake, whole soybeans, soybean meal, and a mixture of defatted soybeans and wheat. The above mixture is a raw material for producing soy sauce cake, and specifically, a mixture of crushed roasted wheat and water that has been sprinkled with defatted soybeans and autoclaved. Soy sauce cake (O-s) is obtained by inoculating the above mixture with A. oryzae S-03. Soybean koji (Od) is a koji obtained by inoculating whole soybeans with water and autoclaving them with A. oryzae S-03. Samples extracted from these straws with twice the amount of water and then lyophilized were used as samples. What was just made to absorb water in a whole soybean and autoclaved was also used.

Each sample dissolved in 100 mg / mL water was sterilized by filtration to prepare untreated, heat-treated at 100 ° C. for 10 minutes, and treated with pepsin or trypsin. A paper disk soaked with these simultaneously was placed on a plate seeded with Pg and then cultured at 37 ° C. for 48 hours to examine the growth inhibitory activity of each sample against Pg.

The mixture of defatted soybean and wheat had no growth inhibitory activity against Pg, but soy sauce cake (Os) and soybean cake (Od) had growth inhibitory activity against Pg (results) Is not shown). Furthermore, the growth inhibitory activity against Pg in soy sauce lees (Os) and soybean lees (Od) was not lost by trypsin treatment, but was lost by pepsin treatment or heat treatment (results not shown). Thus, soy sauce cake (Os) and soybean meal (Od) contain substances that inhibit the growth of Pg, and it is assumed that the active substances are proteins.

[3. Growth-inhibiting active substance for Pg]
Subsequently, 200 mL of a soy sauce cake (O-s) stock solution was extracted twice with three times the amount of chloroform. Subsequently, the obtained chloroform phase was dehydrated and concentrated, and subjected to a bioassay as a chloroform fraction. At the same time, the obtained aqueous phase was extracted twice with ethyl acetate, the obtained ethyl acetate phase was dehydrated and concentrated, and subjected to bioassay as an ethyl acetate fraction and the remaining aqueous phase as a water fraction. Further, a thin layer formed between the aqueous phase and the ethyl acetate phase was used as an intermediate layer for bioassay.

As a result, a blocking circle was formed by the chloroform fraction, the ethyl acetate fraction, and the water fraction, but the blocking circle by the chloroform fraction and the ethyl acetate fraction was very large (results not shown). Thus, the Pg growth inhibitory active substance in soy sauce cake (O-s) is contained in the chloroform fraction and the ethyl acetate fraction.

An attempt was made to purify to a single substance using the above fraction. A bioassay using the W83 strain (type IV Pg distributed from Hiroshima University School of Dentistry, Pediatric Dentistry) for the substance in the band indicated by the arrow in the figure by developing the chloroform fraction and the ethyl acetate fraction with TLC. As a result, a blocking circle was confirmed in the ethyl acetate fraction (results not shown). Therefore, a large amount of this substance was produced and subjected to NMR analysis.

[4. Inhibition of activity against toxin produced by Pg]
The inhibitory activity against the action of gingipain, which is a cysteine protease produced by Pg and exhibits trypsin-like activity, was examined. Extract from koji or fermentation raw material (4 mg / mL or 10 mg / mL) together with Z-His-Glu-Lys-MCA (purchased from Peptide Institute), reaction solution (50 mM sodium acetate (pH 6.8), 100 mM In NaCl, 10 mM DTT) at 37 ° C. for 2 minutes. Next, 100 μg / mL gingipaine solution was added to the reaction solution, and the mixture was reacted at 37 ° C. for 10 minutes. The fluorescence of aminocoumarin produced by this reaction was measured at 355/460 nm. MCA is 4-methyl-7-aminocoumarin, and Z-His-Glu-Lys-MCA is a substrate for Lys-gingipain. As a result, it was found that soybean meal and its extract, and soy sauce cake raw material and its extract strongly inhibit the activity of gingipain (FIG. 2).

[5. Further antibacterial activity)
The antibacterial activity of the koji extract against Candida albicans was examined. After placing a paper disc soaked with water extract (100 mg / mL) of various koji produced using A. sojae No. 9 on a plate seeded with Candida albicans IFO 01385, the temperature was changed to 37 ° C. In each sample, the growth inhibitory activity against Candida was examined (FIG. 3). When producing defatted soybean meal, the defatted soybean was sterilized at 120 ° C. for 30 minutes and then inoculated with conidia of Aspergillus oryzae. Moreover, all the extracts used what performed the heat processing for 10 minutes at 100 degreeC. As shown, the soy sauce koji raw material did not have anti-Candida fungi activity, but the defatted soybean koji or soy sauce koji extract was found to have high anti-Candida fungus activity.

Furthermore, the antibacterial activity of Helicobacter pylori was examined for persimmon or an extract of the persimmon. It is said that H. pylori is currently infected by about 50% of Japanese people, especially 70% of people over the age of 50. H. pylori infection causes gastric and duodenal ulcers, and also causes gastric cancer. Has also been found to be deeply involved.

A paper disc impregnated with water extract (100 mg / mL) of various cocoons produced using A. jasojae 9 No. 9 is placed on a plate seeded with Helicobacter pylori (an isolate from a patient at Hiroshima University Hospital). After placement, each sample was cultured at 37 ° C. for 72 hours to examine the growth inhibitory activity of each sample against H. pylori (FIG. 4). In addition, all each extract used what performed the heat processing for 10 minutes at 100 degreeC. As shown, all of the koji extracts were found to have anti-H. Pylori activity as high as the soy sauce koji raw material.

[6. (Summary)
Table 1 shows the results of examining each antibacterial activity of koji or fermentation raw material. When producing defatted soybean meal, the defatted soybean was sterilized at 120 ° C. for 30 minutes and then inoculated with conidia of Aspergillus oryzae. Moreover, all the extracts used what performed the heat processing for 10 minutes at 100 degreeC. A. oryzae S-03 was used for construction. Further, “+/−” in the table indicates that a slight amount of gingipain inhibitory activity was present.

Thus, the composition of the present invention not only suppresses the growth activity of periodontal disease bacteria but also suppresses the growth activity against Candida and H. pylori. That is, the composition of the present invention can be widely used as an antibacterial composition.

[7. Antibacterial activity in defatted soybean meal extract against periodontal disease bacteria)
Three times the amount of distilled water (60 mL) was added to 20 g of defatted soybean meal obtained using Aspergillus oryzae (A. oryzae S-03), and the mixture was extracted for 1 hour with stirring at room temperature. The extract (30 mL) was sterilized by filtration using a membrane filter (pore size 0.2 μm). This filtrate was freeze-dried to obtain 3.1 g of a powder (defatted soybean meal extract).

The obtained powder is dissolved in distilled water, a solution (solution I) corresponding to 1.8 mg of defatted soybean cake extract is spotted on a filter paper (diameter 8 mm), and periodontal disease Porphyromonas gingivalis (Pg) HNA99 is used as a test bacteria. Antibacterial activity was measured. Pg was anaerobically cultured at 37 ° C. for 72 hours. As a result, a blocking circle having a diameter of 17 mm was formed (FIG. 5a). Prepare 2.5-fold diluted solution (solution II), 5-fold diluted solution (solution III), and 10-fold diluted solution (solution IV) of the above solution, similarly spot the same amount of solution on filter paper, The antibacterial activity of the test bacteria was measured. As a result, inhibition circles of 12 mm, 9.5 mm, and 8.4 mm were formed by the 2.5-fold diluted solution, the 5-fold diluted solution, and the 10-fold diluted solution, respectively. As shown in FIG. 5b, a linear correlation was observed between the weight of defatted soybean meal extract and the minimum inhibitory concentration (MIC) against Pg. The amounts of defatted soybean meal extract contained in the spots of solutions II to IV correspond to 0.72 mg, 0.36 mg and 0.18 mg, respectively.

[8. Oral composition containing defatted soybean meal extract]
According to a known procedure, the following ingredients were mixed in the following proportions (% by weight) to prepare an oral cleaning solution (mouse wash):
Glycerin 5.0
Propylene glycol 3.0
Sodium lauryl sulfate 1.5
Polyoxyethylene (60) hydrogenated castor oil 0.4
Saccharin sodium 0.3
Sodium citrate 0.15
Methyl paraben (preservative) 0.1
Perfume Appropriate amount Distilled water Total 100 (wt%).

35 μL of prepared oral cleaning solution (sample 1), a solution of 2 mg of defatted soybean cake extract in sample 1 (sample 2), 35 μL of prepared 10-fold diluted solution of oral cleaning solution (sample 3), and defatted soybean in sample 3 A solution (sample 4) in which 2 mg of cocoon extract was dissolved was spotted on a filter paper (diameter 8 mm), and antibacterial activity was measured using periodontal disease bacteria Porphyromonas gingivalis (Pg) HNA99 as a test bacterium. Pg was anaerobically cultured at 37 ° C. for 72 hours. The results are shown in FIG.

From the results of Samples 1 and 3, the prepared oral cleaning solution has antibacterial activity against Pg, but loses antibacterial activity against Pg when diluted 10 times. And the antibacterial activity with respect to Pg by a defatted soybean meal extract was confirmed from the result of samples 3 and 4. Further, from the results of Samples 1 and 2, even when the defatted soybean meal extract was contained in the oral cleaning liquid, the antibacterial activity against Pg was not inhibited by the components of the oral cleaning liquid. Thus, an oral composition containing a defatted soybean meal extract is sufficiently practical.

[9. Antibacterial activity in fruit juice extract derived from defatted soybean meal against H. pylori
Three times the amount of distilled water or various fruit juices (3 mL) was added to 1 g of defatted soybean meal obtained using Aspergillus oryzae (A. oryzae S-03), and the mixture was extracted for 1 hour with stirring at room temperature. The extract was sterilized by filtration using a membrane filter and used as an antibacterial activity sample. The fruit juice was used after adjusting commercially concentrated fruit juice to 1-fold (100% reduction) to pH 6.0. Moreover, since the fruit juice has a higher viscosity than distilled water, a membrane filter having a pore size of 0.45 μm was used.

35 μL of each antibacterial activity sample was spotted on a filter paper having a diameter of 8 mm, and the antibacterial activity was measured using Helicobacter pylori as a test bacterium. The results are shown in FIG.

Banana, grapes, dates, peaches, siquasa, oranges, pineapples, and plums did not show a growth inhibition circle against H. pylori, but the extract of defatted soybean meal extracted from these juices was a marked growth inhibition circle. Formed. However, almost no anti-pylori active substance was observed in the defatted soybean meal extract extracted with Unshu mandarin or pomegranate juice. An extract of defatted soybean meal extracted with distilled water was used as a control (the growth inhibition circle diameter for H. pylori was 12 mm). Extracts of defatted soybean meal extracted with banana, grape, dates, peach, shiquasa and orange juices form a growth inhibition circle of 15 to 17 mm, and extraction with these juices yields anti-pylori active substances quantitatively. It was effective. It was also found that the pomegranate juice itself contained an anti-pylori active substance.

The present invention is not limited to the above-described embodiments, and various modifications are possible within the scope shown in the claims, and embodiments obtained by appropriately combining technical means disclosed in different embodiments. Is also included in the technical scope of the present invention.

By using the present invention, it is possible to develop a prophylactic or therapeutic agent for periodontal disease, or a prophylactic or improving agent for bad breath, which is more effective than before. Moreover, if this invention is used, the antimicrobial technique which has a wide antimicrobial spectrum can be provided.

Claims (22)

1. An antibacterial composition containing cocoon or an extract of the cocoon.
2. The composition according to claim 1, further comprising a fermentation raw material for obtaining the koji or an extract of the fermentation raw material.
3. The composition according to claim 1 or 2, wherein the fermentation raw material for obtaining the koji is selected from the group consisting of soybeans, defatted soybeans, wheat, rice, and soy sauce koji materials.
4. The composition according to any one of claims 1 to 3, wherein the koji is produced using koji molds selected from the group consisting of A. oryzae, A. sojae, A. kawachii and A. awamori.
5. The composition according to claim 1 or 2, wherein the fermentation raw material for obtaining the koji is selected from the group consisting of soybeans, defatted soybeans and soy sauce koji materials, and the koji is A. ooryzae.
6. The composition according to any one of claims 1 to 5, wherein the extract of koji is a water extract.
7. The composition according to any one of claims 2 to 6, wherein the extract of the fermentation raw material is a water extract.
8. The composition according to any one of claims 1 to 7, which is used for application in the oral cavity.
9. The composition according to claim 8, which is used for treating periodontal disease.
10. The composition according to claim 8 or 9, which is used for suppressing the growth of Candida.
11. The composition according to any one of claims 8 to 10, which is used for suppressing the growth of Helicobacter pylori.
12. The composition according to any one of claims 1 to 7, which is a quasi-drug as an external preparation.
13. The composition according to any one of claims 1 to 7, which is used for makeup.
14. A quasi-drug comprising the composition according to any one of claims 1 to 13.
15. A food comprising the composition according to any one of claims 1 to 11.
16. A pharmaceutical comprising the composition according to any one of claims 1 to 11.
17. A method for treating periodontal disease, comprising a step of bringing the composition according to any one of claims 1 to 9 into contact with a causative agent of periodontal disease in the oral cavity of a subject.
18. The method according to claim 17, further comprising the step of supplying the composition according to any one of claims 1 to 9 into the oral cavity of the subject.
19. A method for treating oral candidiasis, comprising a step of bringing the composition according to any one of claims 1 to 10 into contact with Candida in the oral cavity of a subject.
20. The method according to claim 19, further comprising the step of supplying the composition according to any one of claims 1 to 10 into the oral cavity of the subject.
21. A method for inhibiting the growth of Helicobacter pylori, which comprises the step of bringing the composition according to any one of claims 1 to 11 into contact with H. pylori in the stomach of a subject.
22. The method according to claim 21, further comprising the step of supplying the composition according to any one of claims 1 to 11 into the stomach of the subject.

Images