KR102782163B1 - Composition for prevention or treatment of oral disease comprising punicalagin - Google Patents

Abstract

The present invention relates to a composition for preventing or treating oral diseases, including punicalagin, and more specifically, to a quasi-drug composition having an effect of preventing or improving toothache, sensitivity, and bad breath. The composition of the present invention has excellent antibacterial activity against bacteria causing dental caries and periodontal disease, and suppresses PGE2, a pain and inflammation marker, in a concentration-dependent manner, and has excellent effects of inhibiting gingivitis formation, suppressing sensitivity symptoms, and eliminating bad breath, and thus has high utility as a quasi-drug composition for preventing or improving at least one oral disease selected from the group consisting of toothache, sensitivity, and bad breath. In addition, the composition of the present invention can be used as a pharmaceutical composition and a food composition.

Description

Composition for prevention or treatment of oral disease comprising punicalagin

The present invention relates to a composition for preventing or treating oral diseases containing punicalagin, and more specifically, to an over-the-counter pharmaceutical composition having the effect of preventing or improving toothache, sensitivity, and bad breath. In addition, the present invention relates to a pharmaceutical composition containing punicalagin, a food composition, and an oral composition containing punicalagin.

Healthy teeth are an important factor in determining the quality of life, so much so that they are called one of the five blessings. Among oral diseases, dental caries and periodontal disease are diseases with high incidence worldwide, causing various clinical symptoms such as pain, impaired chewing function, destruction of periodontal tissue, bad breath, and sensitivity, and are known to be the main factors leading to tooth loss. In addition, the causative factors of oral diseases are increasing due to changes in eating habits.

It is known that there are approximately 600 to 800 kinds of microorganisms in the human mouth, and these microorganisms are controlled by enzymes such as lysozyme secreted in saliva. However, the oral environment rich in nutrients and moisture is a good condition for the growth of microorganisms, and the tongue and dental plaque provide excellent habitats for microorganisms. Some of these microorganisms are opportunistic pathogens, causing diseases such as dental caries, periodontal disease, and dentin hypersensitivity, as well as bad breath.

Antimicrobial agents are a method of suppressing the growth of oral pathogens that reside in dental plaque and cause dental caries, periodontal disease, bad breath, and tooth sensitivity. Various types of antimicrobial agents, including antibiotics with bactericidal and bacteriostatic effects on oral pathogens, have been developed as agents for suppressing and treating dental caries, periodontal disease, and pulp and periapical infections.

However, antibiotics have the disadvantage that they are difficult to use long-term because they can cause systemic side effects on our bodies, as well as the emergence of resistant bacteria and superinfection in the oral cavity, and can only be used as a treatment. In addition, antibacterial agents used in mouthwashes include Sangquinarine, Listerine, Peroxide, and Chlorhexidine. Sangquinarine has an unclear effect on bacteria in the oral cavity, an even more unclear effect on periodontal disease, and is expensive. Listerine is mainly composed of alcohol, so it has a slight bacteriostatic effect, but it only has a temporary effect in the oral cavity, and it has the disadvantage that it can have a harmful effect on tissues when used long-term. In addition, pyroxide, which has recently been added for whitening effects, is toxic to bacteria, but it is also toxic to human tissue, so there are safety issues, and occasionally, bacteria resistant to pyroxide appear. Chlorhexidine is also known to be the best agent among those known to date for the prevention and treatment of periodontal disease as well as for the inhibition of dental plaque formation, but it has the problems of causing tissue irritation, tissue discoloration and degeneration, and especially showing side effects such as a strong irritating taste and a strong odor, and it can induce dysbiosis and is carcinogenic, so it is restricted for use by pregnant women, and so it has the disadvantage of not being able to be used for long-term treatment or especially preventive purposes.

Meanwhile, punicalagin is a phenol compound with molecular formula C ₄₈ H ₂₈ O ₃₀ and molecular weight 1084.71. Pomegranate ( Punica granatum ) and is known to be soluble in solvents such as water and methanol.

Against this backdrop, the inventors of the present invention conducted research to derive a natural material that is effective in preventing and improving oral diseases such as dental caries, periodontal disease, toothache, sensitivity, and bad breath, without concerns about the above-mentioned side effects. As a result, it was found that punicalagin, a natural composition that is safe and can be used with confidence, exhibits an antibacterial effect against bacteria that cause dental caries and periodontal disease, a PGE2 inhibitory effect, a gingivitis formation inhibitory effect, a sensitivity inhibitory effect, and a bad breath elimination effect, and thus a pharmaceutical composition, a food composition, and an oral composition utilizing the same were completed.

Korean Patent Publication No. 10-2015-0050239

Fouad Amr A, et al. Punicalagin alleviates hepatotoxicity in rats challenged with cyclophosphamide, Vol. 45 158p ~ 162p, 2016.

The purpose of the present invention is to provide an over-the-counter pharmaceutical composition for preventing or improving oral diseases, which contains punicalagin or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating oral diseases, comprising punicalagin or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a food composition for preventing or improving oral diseases, comprising punicalagin or a food-wise acceptable salt thereof as an effective ingredient.

Another object of the present invention is to provide an oral composition comprising punicalagin or an acceptable salt thereof as an active ingredient.

In order to solve the above-mentioned problems and achieve the purpose of the present invention, the present invention provides an over-the-counter pharmaceutical composition for preventing or improving oral diseases, which comprises punicalagin or a pharmaceutically acceptable salt thereof as an active ingredient.

In another aspect of the present invention, a pharmaceutical composition for preventing or treating oral diseases is provided, comprising punicalagin or a pharmaceutically acceptable salt thereof as an active ingredient.

As another aspect of the present invention, a food composition for preventing or improving oral diseases is provided, comprising punicalagin or a food-wise acceptable salt thereof as an effective ingredient.

In another aspect of the present invention, an oral composition comprising punicalagin or an acceptable salt thereof as an active ingredient is provided.

The present invention provides a pharmaceutical composition comprising punicalagin or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, “punicalagin” is represented by the chemical formula 1, has a molecular formula of C ₄₈ H ₂₈ O ₃₀ and a molecular weight of 1084.71, and is derived from pomegranate ( Punica It is an organic compound that can be separated from phenol series such as granatum .

The above pomegranate is the fruit of the pomegranate tree, 6~8cm in diameter, round in shape, and covered with a hard, yellowish peel. Both the fruit and the peel are good for preventing high blood pressure and arteriosclerosis, and are effective for gynecological diseases and boils. It is especially effective for dysentery, and contains volatile alkaloids, so it is used as a parasite and tapeworm extermination medicine. The juice is bright in color, and is used to make fruit wine or to make concentrated juice to make drinks or snacks.

The present invention is not particularly limited to a method for obtaining punicalagin, and may be chemically synthesized by a method known in the art, or a commercially available material may be used.

The punicalagin of the present invention may exist in a solvated form as well as an unsolvated form. The punicalagin of the present invention may exist in a crystalline or amorphous form, and all such physical forms are included within the scope of the present invention.

The present invention provides an over-the-counter pharmaceutical composition for preventing or improving oral diseases, comprising the punicalagin or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, specifically, the present invention can provide an over-the-counter pharmaceutical composition for preventing or improving at least one oral disease selected from the group consisting of toothache, sensitivity, and bad breath, comprising the punicalagin or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, “oral disease” refers to various diseases occurring in the oral region, and the oral region refers to the space inside the mouth connecting the front lips to the back palate and the pharynx. In the present invention, the oral disease is a concept that includes all diseases occurring in the oral cavity regardless of their symptoms, and non-limiting examples of the oral disease include dental caries, periodontal disease (periodontitis or gingivitis), toothache, sensitivity, bad breath, etc.

In the present invention, “dental cavities” refers to an infectious disease caused by bacteria living on the tooth surface, and is also called tooth decay, showing symptoms of erosion and loss of the tooth hard tissue. Specifically, the milky white, translucent, and hard substance that covers the surface of the crown of the tooth and protects the tooth dentin is called tooth enamel or enamel, and refers to a cavity caused by damage to the tooth enamel due to acid generated when sugar, starch, etc. are decomposed by oral pathogens living in the mouth. The main cause of dental caries is dental plaque, a bacterial film formed on the surface of teeth. Saliva forms a thin, sticky film on the teeth, and oral microorganisms such as Streptococcus sobrinus and Streptococcus mutans , a type of streptococcus, attach to it to form a biofilm, creating dental plaque, which is further thickened by Fusobacterium . The dental caries of the present invention includes all types of bacteria that cause dental caries, but specifically, it may include at least one selected from the group consisting of Streptococcus mutans , Streptococcus sanguinis , Streptococcus sobrinus , Streptococcus ratti, Streptococcus criceti , Streptococcus anginosus , and lactic acid bacteria, and more specifically, it may be Streptococcus mutans.

The above Streptococcus mutans is a type of streptococcus, a gram-positive bacterium, and is also called a cariogenic bacterium. The above Streptococcus mutans only proliferates on the surface of teeth, and the baby teeth are not fully developed until around 30 months after birth. Therefore, it is difficult for the mutans bacteria to proliferate until this period, as they do after infancy. If the child is not infected with the mutans bacteria from the mouths of adults during infancy and other oral bacteria settle in the mouth, it is difficult for the mutans bacteria to newly enter the already balanced oral ecosystem, and the probability of getting caries throughout life after infancy is significantly reduced. If the causative bacteria have already been infected through the mouths of adults, brushing the teeth frequently and keeping the mouth clean will help prevent caries. However, once the mutans bacteria have settled in the mouth, it is impossible to eradicate the mutans bacteria.

In one embodiment of the present invention, after inoculating a disk containing punicalagin on a medium smeared with Streptococcus mutans, the size of the growth inhibition zone was measured. As a result, it was confirmed that the experimental group containing punicalagin had a growth inhibition zone diameter of 10.0 mm or more, indicating considerably excellent antibacterial activity against Streptococcus mutans (Table 2).

In the present invention, “periodontal disease” is a disease in which inflammation occurs in the gums, periodontal ligaments, and bone tissues supporting the teeth, and is commonly called periodontal disease, and is divided into gingivitis and periodontitis depending on the degree of the disease. It is a relatively mild and quickly recoverable form of periodontal disease, and when it is limited to the gums, i.e., soft tissues, it is called gingivitis, and when this inflammation progresses to the gums and gum bone, it is called periodontitis. There is a V-shaped gap between the gingiva (gum) and the teeth, and when oral pathogens attack the part below the gum line of this groove (sulcus), they release lipopolysaccharide (LPS), which is an inflammatory stimulus, and this causes the gums to swell and bleed, thereby generating inflammation, and thus damaging the periodontal ligament and adjacent tissues. When periodontitis progresses, it damages the periodontal ligament and even the alveolar bone, and eventually causes tooth loss. Nutritional deficiencies such as protein and vitamins, hormonal disorders such as pregnancy or diabetes, smoking, and acquired immunodeficiency syndrome (AIDS) can worsen the disease. In addition, dental plaque and tartar can be cited as other causes of periodontal disease. The periodontal disease of the present invention includes all types of bacteria that cause periodontal disease, but specifically, Actinobacillus actinomycetemcomitans. actinomycetemcomitans ), Porphyromonas gingivalis , Tannerella forsythia , and Treponema denticola The bacteria may be at least one selected from the group consisting of Porphyromonas denticola and Fusobacterium nucleatum , and more specifically, Porphyromonas gingivalis.

The above Porphyromonas gingivalis is a type of Bacteroidete flexible bacteria, a gram-negative bacterium, and an anaerobic bacterium. The above Porphyromonas gingivalis is found in the oral cavity where periodontal disease occurs, and is also found in the upper gastrointestinal tract, respiratory tract, and colon. In patients with chronic periodontal disease, collagen is decomposed by the collagenase enzyme of the above bacteria, and the above bacteria can invade gingival fibroblasts and survive even under a considerable concentration of antibiotics. In addition, the above bacteria can invade gingival epithelial cells and survive for a long time.

In one embodiment of the present invention, after inoculating a disk containing punicalagin on a medium smeared with Porphyromonas gingivalis, the size of the growth inhibition zone was measured. As a result, it was confirmed that the experimental group containing punicalagin had a growth inhibition zone diameter of 10.0 mm or more, indicating considerably excellent antibacterial activity against Porphyromonas gingivalis (Table 2).

In addition, in one embodiment of the present invention, a clinical trial was conducted by selecting experimental subjects with gingivitis, and as a result, it was confirmed that the experimental group using the experimental toothpaste containing the punicalagin had an excellent gingivitis suppression effect and that the gingivitis suppression effect was sustained as normal bleeding was maintained even after 6 months (Table 4).

In the present invention, “toothache” refers to pain in the teeth when eating sweet food, or very cold or hot food, etc., and generally includes not only pain in the teeth themselves, but also pain in the periodontal tissue that supports the teeth to the jawbone. Pain usually occurs when chewing, the gums swell, and a foul-smelling discharge is produced. Toothache shows slightly different pain depending on the cause of the disease. Specifically, pain due to dental caries is not painful in the early stages, but gradually progresses and pain occurs when the nerves inside the teeth are deeply decayed. In the case of impacted teeth, pain is caused by inflammation of the tissues around the teeth. When the teeth are broken or cracked, when they come into contact with cold food or are bitten hard, the teeth crack and stimulate the nerves, causing pain. In the case of pulpitis, pain is felt when cold food comes into contact in the early stages, and when it progresses further, pain is felt when hot food is consumed. When the inflammation progresses and the pulp tissue dies, there is no response to cold or hot things, and pain occurs due to inflammation of the periapical region (the tip of the tooth root).

In one embodiment of the present invention, as a result of confirming the effect of inhibiting PGE2, known as a pain and inflammation marker, the experimental group including the punicalagin showed the effect of inhibiting PGE2 in a concentration-dependent manner, confirming that a composition including punicalagin can be used for the purpose of preventing or improving toothache (Fig. 1).

In the present invention, “sensitive dentine” refers to hypersensitive dentine, and sensitive dentine symptoms refer to dentine hyperesthesia. Sensitive dentine symptoms are felt to be sensitive when exposed dentine part of teeth comes into contact with cold air or irritating food, and 60 to 98% of adults with periodontal disease show symptoms. Sensitive dentine symptoms can appear when teeth are worn toward the gums due to incorrect brushing habits, excessive occlusal force, acid dissolution, poor oral hygiene, periodontal treatment, restorative treatment, or when teeth are dissolved due to acidification. The fundamental cause of sensitive dentine symptoms is that many tubules in the dentine of teeth are exposed to the outside, and due to exposure, all stimuli are transmitted as they are to the nerves in the pulp, causing the same stimuli to react more sensitively than usual and cause pain. Sensitivity symptoms can range from mild symptoms to severe and persistent pain, and since teeth do not regenerate due to their nature, taking painkillers or anti-inflammatory drugs is not a fundamental solution to the sensitivity phenomenon. Sensitivity symptoms can appear all over the teeth, or can be limited to specific areas such as the upper or lower jaw, or the right or left. The most common areas vary depending on the cause, but are mainly the canines and small molars, and the area where the pain is most severe is the cervical region, which is the border between the gums and teeth, in more than 90% of cases.

In one embodiment of the present invention, a clinical trial was conducted by selecting experimental subjects having teeth with dentin hypersensitivity, and as a result, the experimental group using the experimental toothpaste containing punicalagin had an excellent effect in suppressing dentin symptoms, confirming that a composition containing punicalagin can be used for the purpose of preventing or improving dentin sensitivity (Table 6).

In the present invention, “bad breath” refers to an odor originating from the oral cavity and adjacent organs, and 85 to 90% of bad breath originates from the oral cavity, and particularly from the back of the tongue. The main components of bad breath are volatile sulfur compounds, and 90% of the total amount of volatile sulfur compounds is hydrogen sulfide produced from cysteine, and methyl mercaptan and dimethyl sulfide produced from methionine. These components are mainly produced by protein enzymes secreted by anaerobic bacteria, and the back of the tongue is the most important habitat. This area is not well cleaned by saliva, and there are many small depressions, so it becomes a place where bacteria continuously live. The production of volatile sulfur compounds by anaerobic bacteria is the most important cause of bad breath, but it is also caused by oral diseases such as dental caries, periodontitis, and dry mouth. Many types of anaerobic bacteria are involved in causing bad breath, and a non-limiting example of a bad breath-causing bacteria is Porphyromonas gingivalis , which secretes many types and quantities of enzymes.

In one embodiment of the present invention, a clinical trial was conducted on experimental subjects without dental caries, and as a result, the experimental group using the experimental toothpaste containing punicalagin had an excellent bad breath removal effect, confirming that a composition containing punicalagin can be used for the purpose of preventing or improving bad breath (Table 7).

The quasi-drug composition of the present invention may include an oral quasi-drug. The ingredients included in the quasi-drug composition of the present invention may include, in addition to the effective ingredient, ingredients commonly used in oral quasi-drug compositions, and may include, for example, an abrasive, a humectant, a binder, a foaming agent, a sweetener, a preservative, a medicinal ingredient, a flavoring agent, a pigment, a solvent, a whitening agent, a solubilizer, or a pH adjuster.

The oral pharmaceutical composition of the present invention can be manufactured in any formulation commonly manufactured in the art, and may have formulations such as, but not limited to, toothpaste, mouthwash, mouthwash, gum, candy, mouth spray, oral ointment, oral varnish, mouthwash, and gum massage cream.

As an example, when the oral pharmaceutical composition of the present invention is in the form of a toothpaste, it may include a wetting agent, an abrasive, a binder, a foaming agent, a flavoring agent, a sweetener, a coloring agent, a preservative, a medicinal ingredient, a solvent, a pH regulator, etc.

The above humectant is used to make the powder in the toothpaste ingredients into a paste and prevent the toothpaste from hardening in the air. Glycerin, sorbitol, propylene glycol, polyethylene glycol, etc. may be used alone or in combination of two or more, in an amount of 1 to 60 wt%, specifically 10 to 50 wt%, of the total weight of the composition.

The foaming agent above increases the cleaning effect by spreading the toothpaste in the oral cavity and acts as a surfactant to wash away oral contamination. Surfactants such as sodium lauryl sulfate, sodium lauryl sarcosinate, sodium alkyl sulfosuccinate, and sucrose fatty acid ester may be used alone or in combination of two or more, in an amount of 0.5 to 10 wt%, specifically 0.5 to 5 wt%, of the total weight of the composition.

The above binder prevents separation between the powder and liquid components in the toothpaste, and can be used alone or in combination of two or more of cellulose derivatives such as sodium carboxymethyl cellulose, methyl cellulose, and hydroxy propyl cellulose, and sodium alginate, carrageenan, and xanthan gum, in an amount of 0.1 to 5 wt%, specifically 0.3 to 2 wt%, of the total weight of the composition.

The above abrasive removes attachments on the tooth surface without damaging the tooth surface and allows the tooth to have its original gloss. Calcium carbonate (CaCO ₃ ), dibasic calcium phosphate (CaHPO ₄ , CaHPO ₄ 2H ₂ O), anhydrous silicic acid (SiO ₂ 2H ₂ O), aluminum hydroxide (Al(OH) ₃ ), potassium pyrophosphate, magnesium carbonate, etc. may be used alone or in combination of two or more thereof, in an amount of 1 to 60 wt%, specifically 10 to 50 wt%, of the total weight of the composition.

The above flavoring agent is intended to provide a refreshing feeling and odor to the toothpaste and improve the feeling of use. Peppermint oil, spearmint oil, menthol, etc. may be used alone or in a mixture of two or more types, and may be used in an amount of 1 to 60 wt%, specifically 0.01 to 5 wt%, of the total weight of the composition.

The above sweetener is used to remove the unpleasant taste caused by the toothpaste raw material and to improve the refreshing feeling. Saccharic acid, aspartame, xylitol, licorice acid, etc. can be used alone or in combination of two or more types, in an amount of 1 to 60 wt%, specifically 0.01 to 5 wt%, of the total weight of the composition.

The effective ingredients are for the prevention of dental caries, periodontal disease, toothache, sensitivity, and bad breath removal, and may include fluoride, zinc chloride, chlorhexidine, aminocaproic acid, tranexamic acid, cetylpyridium chloride, pyridoxine chloride, triclosan, tocopheryl acetate, sodium monofluorophosphate, etc., either alone or in combination of two or more. In the present invention, punicalagin may be used as an additional effective ingredient.

The oral pharmaceutical composition of the present invention may be used alone or in combination, or may be used in combination with another oral pharmaceutical composition other than the present invention.

In another aspect of the present invention, a pharmaceutical composition comprising the punicalagin or a pharmaceutically acceptable salt thereof as an active ingredient is provided. In addition, specifically, the present invention can provide a pharmaceutical composition for preventing or treating at least one oral disease selected from the group consisting of toothache, sensitivity and bad breath, comprising the punicalagin as an active ingredient. Punicalagin, oral disease, toothache, sensitivity and bad breath are as described above.

The pharmaceutical composition of the present invention can be formulated in the form of tablets, pills, powders, granules, capsules, suspensions, oral solutions, emulsions, syrups, aerosols, sterile injection solutions, etc. according to conventional methods for preventing and treating oral diseases.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations can be prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, syrups, etc., and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives can be used.

Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, suppositories, etc. Non-aqueous solutions and suspensions may include vegetable oils such as propylene glycol, polyethylene glycol, olive oil, and injectable esters such as ethyl oleate.

In addition, the pharmaceutical composition of the present invention may further comprise a carrier, excipient or diluent. As the carrier, excipient or diluent, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, hydroxypropyl methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil such as silicon dioxide, etc. may be used.

The specific dosage of the pharmaceutical composition according to the present invention can be variously selected by those skilled in the art according to factors such as the formulation method, the patient's condition and weight, the patient's sex, age, degree of disease, drug form, administration route and period, excretion rate, reaction sensitivity, etc., and the dosage and frequency of administration do not limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, and humans through various routes. All modes of administration can be envisaged, and for example, administration can be by oral, intravenous, intramuscular, or subcutaneous injection.

In another aspect of the present invention, the present invention provides a food composition comprising the punicalagin or a food-wise acceptable salt thereof as an active ingredient. In addition, specifically, the present invention can provide a food composition for preventing or improving at least one oral disease selected from the group consisting of toothache, sensitivity, and bad breath, comprising the punicalagin as an active ingredient. Punicalagin, oral disease, toothache, sensitivity, and bad breath are as described above. The food composition can be used in the form of a health functional food, but is not limited thereto.

The punicalagin or a food-wise acceptable salt thereof included in the food composition of the present invention may be included in the form of an animal or plant containing punicalagin, an extract thereof, a fraction thereof, or a processed product thereof. In addition, the composition may include a food-wise acceptable food additive in addition to the effective ingredient.

In the present invention, "food supplement additive" means a component that can be added to food as an auxiliary, and can be appropriately selected and used by those skilled in the art as added in the manufacture of health functional foods of each formulation. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc., but the types of food supplement additives of the present invention are not limited by the above examples.

The food composition of the present invention may include a health functional food. In the present invention, the term "health functional food" refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and pills using raw materials or ingredients having useful functionality for the human body. Here, "functionality" means obtaining a useful effect for health purposes such as regulating nutrients for the structure and function of the human body or physiological effects. The health functional food of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art during the manufacturing process. In addition, the formulation of the health functional food can be manufactured without limitation as long as it is a formulation recognized as a health functional food. The food composition of the present invention can be manufactured in various forms of formulations, and unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long period of time by using food as a raw material, and is highly portable, so the health functional food of the present invention can be consumed as a supplement for preventing or improving oral diseases.

There is no limitation on the form that the health functional food of the present invention can take, and can include all foods in the conventional sense, and can be used interchangeably with terms known in the art such as functional food. In addition, the health functional food of the present invention can be manufactured by mixing other appropriate auxiliary ingredients that can be included in foods and known additives according to the selection of a person skilled in the art. Examples of foods that can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and can be manufactured by adding it to juice, tea, jelly, and juice manufactured using the punicalagin according to the present invention as a main ingredient. In addition, foods used as feed for animals are also included.

The quasi-drug composition comprising the punicalagin of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient has an excellent effect in preventing or improving oral diseases. Specifically, the composition of the present invention has excellent antibacterial activity against bacteria causing dental caries and periodontal disease, and suppresses PGE2, a pain and inflammation marker, in a concentration-dependent manner, and has excellent effects in inhibiting gingivitis formation, inhibiting tooth sensitivity symptoms, and eliminating bad breath, and thus has high utility as a quasi-drug composition for preventing or improving at least one oral disease selected from the group consisting of toothache, tooth sensitivity, and bad breath. In addition, the composition comprising the punicalagin of the present invention can be used as a pharmaceutical composition and a food composition.

The following drawings attached to this specification illustrate preferred embodiments of the present invention and, together with the contents of the invention described above, serve to further understand the technical idea of the present invention; therefore, the present invention should not be interpreted as being limited to matters described in such drawings.
Figure 1 shows the results of confirming the PGE2 inhibitory effect by treating macrophages with LPS (control group) and LPS and punicalagin (experimental group), which are inflammatory stimuli, respectively.

Hereinafter, the composition and effects of the present invention will be described in more detail through examples and experimental examples. These examples and experimental examples are only intended to illustrate the present invention, and the scope of the present invention is not limited by these examples and experimental examples.

Experimental example 1 : Dental caries and antibacterial effect against periodontitis-causing bacteria

In order to confirm the preventive or therapeutic effect of the above punicalagin on dental caries and periodontitis, an antibacterial activity experiment was performed. In order to confirm the effect of inhibiting the growth of oral pathogens, Streptococcus mutans, a representative bacterium causing dental caries, and Porphyromonas gingivalis, a representative bacterium causing periodontal disease, were tested. The antibacterial activity test was conducted using the paper disk assay method using gingivalis .

After increasing the activity of each of the above oral pathogens under the optimal culture conditions of Table 1 below, they were cultured for 4 to 6 hours in the optimal medium for each bacteria to adjust the turbidity of the culture solution to Macfarland turbidity No. 0.5 (1.5 X 10 ⁸ ), and 0.1 ml of each oral pathogen was evenly spread on the plate medium. Thereafter, the punicalagin was inoculated onto the sterilized paper disc at a concentration of 10 mg/disc and absorbed and dried for 1 hour. Then, after culturing for 24 to 48 hours at the optimal temperature for each of the above oral pathogens, the size of the growth inhibition zone (diameter mm) was measured, and the results are shown in Table 2 below.

Strain Optimal conditions Gram stain
(Gram staining) Strain name temperature badge characteristics Gram (+) Streptococcus mutans 37 BHI Facultative anaerobic Gram (-) Porphyromonas gingivalis 37 TSA Hemin Menadione Badge Anaerobic

Results of measurement of antibacterial activity of punicalagin against oral pathogens Growth inhibition ring diameter (mm) Streptococcus mutans Porphyromonas gingivalis Unprocessed - (Growth is not inhibited) - (Growth is not inhibited) Punicalagin treatment 10.6 10.3

As shown in Table 2 above, compared to the untreated group that was not treated with punicalagin, the group treated with punicalagin showed significantly superior antibacterial activity against Streptococcus mutans and Porphyromonas gingivalis, with a growth inhibition zone diameter of approximately 10.0 mm for the two types of oral pathogens.

Experimental example 2: Effect of inhibiting gingivitis formation

In order to confirm the efficacy of the above punicalagin in preventing or treating gingivitis, a clinical trial was conducted by selecting a group of experimental subjects.

First, a control toothpaste was made using sodium carboxymethyl cellulose, sodium lauryl sulfate, glycerin, colloidal silicon dioxide, silica, sodium cocoyl isethionate, dodicin, and sweeteners, air fresheners, and coloring agents commonly used in the manufacture of toothpaste, and an experimental toothpaste was made by adding 0.01 wt% of the punicalagin to the control toothpaste.

In order to select experimental subjects, a detailed oral examination was performed on gingivitis patients with even teeth and no missing teeth, 30 subjects by gender at 10-year intervals from 30 to 50 years of age. Then, 120 experimental subjects were selected and divided into groups of 60 subjects each, and a clinical trial on the treatment effect of gingivitis was performed as follows.

Specifically, the experimental subjects were divided into a control group and an experimental group, and the control group was instructed to use the manufactured control group toothpaste three times a day, two hours after meals and before going to bed, and the experimental group was instructed to use the manufactured experimental group toothpaste three times a day at the same time. After that, dental floss was performed to score the initial gingivitis index, and the control group was instructed to use the manufactured control group toothpaste and the experimental group was instructed to use the manufactured experimental group toothpaste, and the gingivitis index was examined after 1 week, 1 month, 3 months, and 6 months by performing an oral examination. The gingivitis index was measured by inserting a periodontal probe into the gingival sulcus, burning the area around each tooth without applying force, and measuring the bleeding after 30 seconds, and recording the score according to the criteria shown in Table 3 below, and the results are shown in Table 4.

Gingivitis Index Standard score detail 0 points Bleeding-free state 1 point Normal bleeding condition 2 points Onboard bleeding condition 3 points Interdental triangular bleeding condition 4 points Bleeding condition of entire gums

Anti-gingival inflammation effect hour Control group Experimental group 0 weeks 1.04 1.03 1 week 1.56 1.05 1 month 2.71 1.06 3 months 2.83 1.08 6 months 2.93 1.09

As shown in Table 4 above, compared to the control group, the experimental group that used toothpaste containing punicalagin maintained a normal bleeding state even after 6 months, indicating that the effect of suppressing gingival inflammation persisted.

Experimental example 3: Pain and inflammation Marker PGE2 Inhibitory effect

In order to confirm the effect of the above punicalagin on preventing or treating toothache, the inhibitory effect of PGE2, known as a pain and inflammation marker, was confirmed.

First, macrophages were seeded at a concentration of 1.5 X 10 ⁵ cells/ml in DMEM medium containing 10% FBS and cultured in a 24-well plate at 37°C, 5% CO ₂ for 24 hours. Then, 1 μg/ml of LPS, an inflammation-inducing stimulus, and punicalagin were treated at various concentrations and cultured for an additional 24 hours. A PGE2 ELISA assay kit (Thermo SCIENTIFIC) was purchased and the supernatant was used to measure and analyze the PGE2 inhibitory effect according to the concentration of punicalagin according to the instructions (Fig. 1).

As shown in Fig. 1, compared to the group treated with only LPS, punicalagin at a concentration of 0.2 ppm had the effect of inhibiting PGE2 by about 60%, and punicalagin at a concentration of 2 ppm had the effect of inhibiting PGE2 by about 80%. Therefore, it was found that punicalagin can be used for the purpose of preventing or treating toothache.

Experimental example 4 : Sirin Inhibitory effect

In order to confirm the preventive or therapeutic effect of the above punicalagin, a clinical trial was conducted by selecting experimental subjects, and the control group toothpaste and experimental group toothpaste used in the clinical trial were each manufactured using the same method as the toothpaste of Experimental Example 2.

The subjects of the experiment were 40 volunteers who agreed to participate in this experiment with dentin hypersensitivity teeth, and the total number of experimental teeth was 80. In addition, among the subjects of the experiment, there were 20 men and 20 women, and their ages ranged from 20 to 50. The subjects were not informed of the contents of the toothpaste, and the total experimental period was 2 weeks.

The experiment was conducted by measuring the reaction of the experimental subjects after applying a temperature stimulus. Before conducting the experiment, the hypersensitive area of the dentin hypersensitivity teeth of each experimental subject was checked in advance, and cold water of about 5℃ was dropped with a dropper on the sensitive area of the tooth, and a score was given according to the criteria shown in Table 5 below. The control group used the control group toothpaste, and the experimental group used the experimental group toothpaste three times a day for two weeks. After two weeks, cold water of about 5℃ was dropped with a dropper again, and a score was given. Statistical processing was performed by testing the stimulation scores before and two weeks after the experiment using a paired Student-t test, and the reaction scores to the temperature stimulus two weeks later are shown in Table 6 below.

Subjects' rating criteria after stimulation score detail 0 points There is no discomfort at all. 1 point A little uncomfortable or cold. 2 points It's very cold. 3 points sick.

The degree of suppression of siren hour Control group Experimental group 0 weeks 2.14±0.13 2.14±0.09 2 weeks 2.13±0.12 1.70±0.20 p>0.05 *p<0.05

※ p>0.05: 95% confidence, no statistically significant difference. *p<0.05: 95% confidence, there is a statistically significant difference.

As shown in Table 6 above, compared to the control group, the experimental group that used toothpaste containing punicalagin showed suppression of tooth sensitivity after two weeks. Therefore, it was found that punicalagin can be used for the purpose of preventing or treating tooth sensitivity.

Experimental example 5: Bad breath removal effect

In order to confirm the effect of the above punicalagin on preventing or treating bad breath, a clinical trial was conducted by selecting experimental subjects, and the control group toothpaste and experimental group toothpaste used in the clinical trial were each manufactured using the same method as the toothpaste of Experimental Example 2.

A cross-over test was conducted on the control group toothpaste and the experimental group toothpaste by selecting 50 men and women without dental caries as the experimental subjects. Commercially available garlic powder was dispersed in water, left for 24 hours, and then diluted so that the Halimeter measurement value was 700 ppb or higher, and the diluted solution was used as a bad breath inducer. The experimental subjects gargled with 15 ml of the diluted garlic powder solution for 30 seconds, and after 1 minute, the degree of bad breath was measured with a Halimeter. The control group used the control group toothpaste, and the experimental group used the experimental group toothpaste, respectively, and brushed their teeth for 30 seconds to 1 minute. The degree of bad breath was measured with a Halimeter 1 minute, 5 minutes, and 30 minutes after brushing, to determine the persistence of bad breath suppression, and the results are shown in Table 7 below.

Bad breath removal effect hour Control group Experimental group 1 minute 54.8% 96.0% 5 minutes 48.1% 89.2% 30 minutes 40.0% 87.1%

As shown in Table 7 above, compared to the control group, the experimental group that used toothpaste containing punicalagin had approximately 96% of bad breath removed after brushing their teeth, and even after 30 minutes of brushing, the bad breath removal rate was over 87%, showing that the bad breath removal effect was far superior to the control group. It was found that punicalagin can be used for the purpose of preventing or treating bad breath.

Claims (9)

A pharmaceutical composition for preventing or improving oral diseases, containing punicalagin or a pharmaceutically acceptable salt thereof as an active ingredient,
A pharmaceutical composition, wherein the oral disease is periodontitis, gingivitis or otitis media.
delete delete delete A pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is for oral use.
In claim 5, the formulation of the non-medical composition is any one selected from the group consisting of toothpaste, mouthwash, mouthwash, gum, candy, mouth spray, oral ointment, oral varnish, mouthwash, and gum massage cream.
A pharmaceutical composition for preventing or treating oral diseases, comprising punicalagin or a pharmaceutically acceptable salt thereof as an active ingredient,
A pharmaceutical composition, wherein the oral disease is periodontitis, gingivitis or otitis media.
A food composition for preventing or improving oral diseases, comprising punicalagin or a food-based acceptable salt thereof as an active ingredient.
A food composition, wherein the oral disease is periodontitis, gingivitis or gingivitis.

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