WO2020060277A1 - Composition for preventing or treating oral diseases or bone diseases - Google Patents

Abstract

The present invention pertains to: a pharmaceutical composition, a food composition, a quasi-drug composition, and a dental composition, which are for preventing or treating oral diseases or bone diseases and contain, as an active ingredient, any one or more selected from the group consisting of a taro extract, a long-tail spleenwort extract, and a slender false brome extract. Specifically, the taro extract, the long-tail spleenwort extract, and the slender false brome extract in the present invention significantly inhibit the activity of the oral disease-causing bacteria, Porphyromonas gingivalis, and significantly inhibit the formation or differentiation of osteoclasts, while having low toxicity due to being derived from natural products, and can thus be useful for preventing or treating oral diseases or bone diseases.

Description

Composition for prevention or treatment of oral disease or bone disease

The present invention relates to a composition for the prevention or treatment of oral diseases or bone diseases comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract, specifically taro extract, tail It relates to a pharmaceutical composition, food composition, quasi-drug composition and dental composition for the prevention or treatment of oral diseases or bone diseases comprising at least one selected from the group consisting of fern extract and forest buckwheat extract.

In osteoclasts, RAW264.7 monocytes of mice are differentiated into multinucleated osteoclasts by a receptor activator of nuclear factor ĸB (RANK) ligand (RANKL). This differentiation process promotes the activity of mitogen-activated protein kinase (MAPK) by binding RANKL from the outside of the cell to RANK, which is a TRAP associated with osteoclast differentiation due to the transcription factor NF-ĸB entering the nucleus. It is possible by increasing the expression of (tartrate-resistant acid phosphatase), MMP-9 (matrix metalloproteinase-9), c-Src tyrosine kinase, etc. The multinuclear osteoclasts formed by this process are able to replace mineralized bone. It serves to absorb. In addition, when RANKL binds to RANK, it promotes the activity of TRAF6 (tumor necrosis factor receptor-associated factor 6), thereby promoting the activity of transcription factors such as MARK, or NF-ĸB, AP-1, NFATc1 (LEE ZH, KIM) HH.Signal transduction by receptor activator of nuclear factor kappa B in osteoclasts.Biochem Biophys Res Commun. 2003 May 30, 305, 211-4). Therefore, blocking the signaling pathway activated by RANKL is recognized as one of the therapeutic approaches in the treatment of diseases associated with osteoclast differentiation.

Osteoblasts cause abnormal destruction and absorption of bone tissue due to imbalance with osteoblasts in the bone, and thereby, osteoporosis, which decreases bone mass and bone density, osteomalacia, where lime is lost from bone, normal Fibrous dysplasia where bone tissue is replaced by fibrous connective tissue and immature bone soju, periodontal disease in which alveolar bone is lost, and rheumatoid arthritis that causes joint destruction and deformation. It is known. As such, there is a close relationship between osteoclast differentiation and periodontal disease and bone disease.

Porphyromonas gingivalis is known to inhibit osteoblast differentiation by host-mediated inflammatory reactions through lipopolysaccharide (LPS) or ciliary mediation (Wilson, 1998; Kadono et al.) 1999), the LPS is a well-known stimulator of the synthesis of inflammatory cytokines or eicosanoids as a pathogenic component present in the outer membrane of the bacterium. It stimulates osteoblasts to osteolytic factors, namely IL-1, IL-6, PGE2, oxidation It is known as a major structure that destroys periodontal tissue by secreting nitrogen. In addition, the differentiation of osteoclasts is directly promoted and activated by the LPS itself, which can ultimately lead to bone loss (Koide et al., 2000; Taubman et al., 2005; Islam et al., 2007; Hou et al., 2013; Teramachi, etc.) 2017). As such, there are a number of data on the effects of periodontal disease-causing strains on the differentiation of osteoblasts or osteoclasts, and there are also therapeutic agents to treat osteoarthritis and periodontal disease. This exists.

Currently, antibiotics and antibacterial agents such as Sangquinarine, Listerine, Peroxide, and Chlorhexidine are widely used as therapeutic agents to treat oral diseases including periodontal disease. However, since antibiotics can induce the appearance of resistant bacteria in the oral cavity and superinfection along with systemic side effects on our body, there is a drawback that long-term use is difficult and can only be used as a therapeutic agent. In addition, raw guinarine has an unclear effect on bacteria in the oral cavity and a more unclear treatment effect on periodontal disease, and also has a disadvantage in that it is expensive. It has a short-term effect, but also has the disadvantage that it may have a harmful effect on tissues when used for a long time. In addition, the recently added pyroxide for the whitening effect is toxic to bacteria, but at the same time it is toxic to human tissues, which is not only a problem in safety, but also sometimes occurs in bacteria resistant to pyroxide. In addition, chlorhexidine is known to be the best among the formulations known to date as a preventive and therapeutic agent for periodontal disease along with inhibition of plaque formation, but it causes irritation to tissues, coloring and degeneration of tissues, and has a particularly strong, irritating and odorous side effect. In addition, there is a problem that cannot be used for a long period of time for the purpose of treatment or prevention, for example, because it can cause myceliacosis and has limited carcinogenicity. Therefore, there is a need to develop a medicine capable of effectively suppressing the growth of strains causing oral diseases without concern for the side effects as described above.

Currently, bisphosphonate-based treatments are widely used to treat bone damage caused by osteoclasts such as osteoporosis. However, in recent years, patients taking bisphosphonate-based drugs have been increasing year by year with various side effects such as osteonecrosis, severe atrial fibrillation, disabling bone or joints, and musculoskeletal pain (Br J Cancer 98: 1736). -1740 (2008)). Therefore, there is a need to develop pharmaceuticals that compensate for the disadvantages of the existing bisphosphonate-based drugs, have low toxicity, and can effectively suppress osteoclast formation and differentiation.

One object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of oral disease or bone disease comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract as an active ingredient. .

Another object of the present invention is to provide a food composition for preventing or improving oral disease or bone disease, including any one or more selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract as an active ingredient. .

Another object of the present invention is to provide a quasi-drug composition for the prevention or improvement of oral disease or bone disease comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract as an active ingredient will be.

Another object of the present invention is to provide a dental composition for the prevention or improvement of oral diseases comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract as an active ingredient.

Under these backgrounds, the present inventors have shown that they are less toxic from natural products, and have studied politely to find compounds that can inhibit the activity of oral disease-causing bacteria and inhibit osteoclast formation or differentiation. Oral disease by remarkably inhibiting the activity of Porphyromonas gingivalis , which is an oral disease-causing bacterium, and at least one selected from the group consisting of extracts and forest buckwheat extracts, and significantly inhibits the formation or differentiation of osteoclasts Or by confirming that it can be useful for the prevention or treatment of bone disease, the present invention was completed.

The composition comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract of the present invention as an active ingredient is derived from a natural product and has low toxicity, but also causes oral disease-causing bacteria, Porphyromonas jinjibali S. ( Porphyromonas gingivalis ) by significantly inhibiting the activity and by significantly suppressing the formation or differentiation of osteoclasts can be useful for the prevention or treatment of oral disease or bone disease.

Figure 1a is a graph showing the cytotoxicity (cell viability) according to the treatment concentration of the taro extract.

1B is a graph showing cytotoxicity (cell viability) according to the treatment concentration of tail fern extract.

Figure 1c is a graph showing the cytotoxicity (cell viability) according to the treatment concentration of the forest buckwheat extract.

Figure 2a is a graph showing the effect of inhibiting the production of nitrogen oxides according to the treatment concentration of the taro extract. The significance test was performed through the student t-test, and the significance # of the LPS-treated group compared to the control group was p <0.001. The significance of the taro extract-treated group compared to the LPS-treated group was p <0.05, ** was p <0.01, and *** was p <0.001.

Figure 2b is a graph showing the effect of inhibiting the production of nitric oxide according to the treatment concentration of the tail fern extract. The significance test was performed through the student t-test, and the significance # of the LPS-treated group compared to the control group was p <0.001. The significance of the tail fern extract-treated group compared to the LPS-treated group was p <0.05, ** is p <0.01, and *** is p <0.001.

Figure 2c is a graph showing the effect of inhibiting the production of nitric oxide according to the treatment concentration of the forest buckwheat extract. The significance test was performed through the student t-test, and the significance # of the LPS-treated group compared to the control group was p <0.001. The significance of the forest buckwheat extract treatment group compared to the LPS treatment group is p <0.05, ** is p <0.01, and *** is p <0.001.

Figure 3a is a graph showing the inhibitory effect of inflammatory cytokines, (a) IL-6 and (b) TNF-α production according to the treatment concentration of the taro extract. The significance test was performed through the student t-test, and the significance # of the LPS-treated group compared to the control group was p <0.001. The significance of the taro extract-treated group compared to the LPS-treated group was p <0.05, ** was p <0.01, and *** was p <0.001.

Figure 3b is a graph showing the inhibitory effect of inflammatory cytokines, (a) IL-6 and (b) TNF-α production according to the treatment concentration of the tail fern extract. The significance test was performed through the student t-test, and the significance # of the LPS-treated group compared to the control group was p <0.001. The significance of the tail fern extract-treated group compared to the LPS-treated group was p <0.05, ** is p <0.01, and *** is p <0.001.

Figure 3c is a graph showing the inhibitory effect of inflammatory cytokines, (a) IL-6 and (b) TNF-α production according to the treatment concentration of the forest buckwheat extract. The significance test was performed through the student t-test, and the significance # of the LPS-treated group compared to the control group was p <0.001. The significance of the forest buckwheat extract treatment group compared to the LPS treatment group is p <0.05, ** is p <0.01, and *** is p <0.001.

Figure 4a is a graph showing the growth inhibitory effect of Porphyromonas gingivalis according to the treatment concentration of the taro extract. The significance test was performed through the student t-test, and the significance * compared to the control group was p <0.05, ** indicates p <0.01, and *** indicates p <0.001.

Figure 4b is a graph showing the growth inhibitory effect of Porphyromonas gingivalis according to the treatment concentration of the tail fern extract. The significance test was performed through the student t-test, and the significance * compared to the control group was p <0.05, ** indicates p <0.01, and *** indicates p <0.001.

Figure 4c is a graph showing the growth inhibitory effect of Porphyromonas gingivalis according to the treatment concentration of the forest buckwheat extract. The significance test was performed through the student t-test, and the significance * compared to the control group was p <0.05, ** indicates p <0.01, and *** indicates p <0.001.

Figure 5a is a graph showing the sustained effect of growth inhibition of Porphyromonas gingivalis according to the control group, fluoride varnish treatment group, fluoride varnish + taro treatment group. The significance test was conducted through the student t-test, and the significance ### of the fluoride varnish treatment group compared to the control group was p <0.001. The significance *** of the fluoride varnish + taro treatment group compared to the fluoride varnish treatment group represents p <0.001.

Figure 5b is a graph showing the sustained effect of growth inhibition of Porphyromonas gingivalis according to the control group, fluoride varnish treatment group, fluoride varnish + tail fern treatment group. The significance test was conducted through the student t-test, and the significance ### of the fluoride varnish treatment group compared to the control group was p <0.001. The significance *** of the fluoride varnish + tail fern treatment group compared to the fluoride varnish treatment group was p <0.001.

Figure 5c is a graph showing the sustained effect of growth inhibition of Porphyromonas gingivalis according to the control group, fluoride varnish treatment group, fluoride varnish + forest wheat treatment group. The significance test was conducted through the student t-test, and the significance ### of the fluoride varnish treatment group compared to the control group was p <0.001. The significance *** of the fluoride varnish + forest cultivation treatment group compared to the fluoride varnish treatment group was p <0.001.

Figure 6 is a photograph showing the TRAP staining results of (a) TRAP-positive cells, and (b) differentiated osteoclasts according to the treatment concentration of the taro extract. The significance test was performed through the student t-test, and the significance * compared to the control group was p <0.05, ** indicates p <0.01, and *** indicates p <0.001.

7 is a photograph showing the results of TRAP staining of differentiated osteoclasts, and (a) a graph showing the number of TRAP positive cells according to the treatment concentration of the tail fern extract. The significance test was performed through the student t-test, and the significance * compared to the control group was p <0.05, ** indicates p <0.01, and *** indicates p <0.001.

8 is a photograph showing the results of TRAP staining of differentiated osteoclasts, and (a) a graph showing the number of TRAP-positive cells according to the treatment concentration of the forest buckwheat extract. The significance test was performed through the student t-test, and the significance * compared to the control group was p <0.05, ** indicates p <0.01, and *** indicates p <0.001.

In order to achieve the above object, one aspect of the present invention is for the prevention or treatment of oral disease or bone disease comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract as an active ingredient Provided is a pharmaceutical composition.

Specifically, the pharmaceutical composition for preventing or treating oral disease or bone disease of the present invention includes an extract of taro, tail fern or forest buckwheat; Or fractions thereof.

The term of the present invention, "Taro ( Colocasia Antiquorum Var. Exculenta )" is a perennial herb belonging to the Cheonnamseong and is also called cultivation . Leaves, stems, etc. are edible, and in general, edible stems are used. It is said that this main name is called vaccinia, it adds the energy of the spleen and detoxification, and has been applied to various skin diseases and burns.

Taro is known to have various pharmacological properties such as fatty liver, diabetes complications, and depression treatment effects, but its prevention, treatment, or improvement effects on oral diseases or bone diseases have not been known.

The term "tail fern ( Asplenium incisum )" is a small fern plant with short rhizomes. Since the outpost has various effects such as detoxification, it uses raw juice for medicinal purposes. Antibacterial activity and bronchodilatory activity against skin disease causative bacteria have been reported.

The tail fern is known to have various pharmacological properties such as hepatitis, tinnitus, and chronic manganese poisoning treatment effects, but the effect of preventing, treating or improving its oral disease or bone disease is not yet known.

The term "forest buckwheat ( Brachypodium sylvaticum )" is a perennial plant belonging to the family Gramineae and forest stalk ( Brachypodium ), inhabiting the southern regions of Korea, especially the coastal islands.

Forest wheat is known to be used as feed for livestock, but its various pharmacological properties, or its prevention, treatment, or improvement effects on oral diseases or bone diseases have not been known.

The present inventors have conducted various studies on substances that show excellent prevention or treatment effects of oral diseases or bone diseases from natural resources existing in nature, as a result of taro extract, tail fern extract or forest buckwheat extract, Porphyromonas, which is an oral disease-causing bacteria. It was confirmed that it can be effectively used for the prevention or treatment of oral diseases or bone diseases by inhibiting the activity of Porphyromonas gingivalis and inhibiting the formation or differentiation of osteoclasts. Through this, it was found that taro extract, tail fern extract or forest buckwheat extract can be used as a pharmaceutical composition for preventing or treating oral disease or bone disease. The composition comprising any one or more selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract having the effect of preventing or treating the oral disease or bone disease has not been known so far, and the inventors It has a very great significance in that it was first developed by.

For the purposes of the present invention, taro, tail fern or forest wheat is not limited to its kind, and can be used cultivated, commercially available, or used without limitation in its source.

In the present invention, the term, "extract" means that any one or more selected from the taro, tail fern or forest wheat is extracted, but is not limited thereto, an extract obtained by the extraction process, a dilution or concentrate of the extract , The dried product obtained by drying the extract, may be included in the extract itself and the extracts of all formulations that can be formed using the extract, such as a refined product or a mixture thereof. In addition, it can be extracted from various organs of natural, hybrid, and varietal plants of the taro, tail fern or forest clover, for example, roots, rhizomes, ground parts, stems, leaves, and fruits.

The taro extract, tail fern extract or forest buckwheat extract of the present invention can be prepared using general extraction methods, separation and purification methods known in the art. The extraction method is not limited thereto, but a method such as immersion extraction, hot water extraction, cold immersion extraction, reflux cooling extraction or ultrasonic extraction may be used.

The taro extract, tail fern extract or forest buckwheat extract may be extracted with water, alcohol having ₁ to ₁ carbon atoms (C ₁ ) to 4 carbon atoms (C ₄ ), or a mixed solvent thereof, but is not limited thereto. Accordingly, since the extraction degree and loss degree of the active ingredient may be different, an appropriate organic solvent may be selected and used. Specifically, it can be extracted using water, an organic solvent, or a mixed solvent thereof. More specifically, ethanol may be used, but is not limited thereto.

In addition, the solvent extract may further include filtering the extract to remove floating solid particles. As an example, the particles may be filtered using cotton, nylon, or the like, an ultrafiltration method, a freezing filtration method, or a centrifugation method may be used, but is not limited thereto.

For concentration of the extract, a method such as concentrated under reduced pressure or concentrated by reverse osmosis may be used.

After the concentration and drying step, freeze drying, vacuum drying, hot air drying, spray drying, vacuum drying, foam drying, high frequency drying, or infrared drying may be used. Optionally, a process of grinding the final dried extract may be further included.

The term "fraction" in the present invention means a result obtained by performing a fraction to separate a specific component or a specific component group from a mixture containing various various components.

The fractionation method for obtaining the fraction in the present invention is not particularly limited as long as it can exhibit the prevention or treatment effect of oral diseases or bone diseases, but may be performed according to methods commonly used in the art. For example, a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing through an ultrafiltration membrane having a constant molecular weight cut-off value, and various chromatography (separation according to size, charge, hydrophobicity or affinity) Chromatography), and combinations thereof.

In the present invention, the type of fractional solvent used to obtain the fraction is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the fractional solvent include polar solvents such as water, distilled water, and alcohol; And non-polar solvents such as hexane, ethyl acetate, chloroform and dichloromethane. These may be used alone or in combination of two or more. When using an alcohol in the fractional solvent, preferably, an alcohol having 1 to 2 carbon atoms (C ₁ ) to 4 (C ₄ ) carbon atoms can be used.

In addition, the extract or fraction may be prepared and used in a dry powder form after extraction, but is not limited thereto.

In the present invention, "oral disease" refers to various diseases that occur in the oral area, and the oral area refers to a space in the mouth that is connected to the pharynx from the anterior lip to the posterior canal. In the present invention, the oral disease is a concept that includes all irrespective of the pathology if the disease occurs in the oral cavity, and non-limiting examples of the oral disease include dental caries, periodontal disease (periodontitis or gingivitis), toothache, and bad breath. And the like.

The term "periodontal disease" of the present invention is also commonly referred to as qichi, and is divided into gingivitis and periodontitis depending on the degree of disease. Gingivitis, a form limited to soft tissues, is a relatively light and fast-recovering oral disease called gingivitis, and periodontitis is the case when such inflammation has progressed to the gingiva and alveolar bones.

There is a V-shaped gap between the gingiva (gingival) and the teeth. It is a periodontal disease in which bacteria attack the lower gingival portion of the gingival sulcus and damage the periodontal ligament and adjacent tissue. As inflammation progresses and more tissue is damaged, the groove develops into a periodontal pocket, and the deeper the periodontitis, the deeper the depth of the periodontal pocket. The periodontal disease is caused by inflammation of the periodontal ligament as the periodontal pocket deepens, and osteoclast differentiation is promoted by lipopolysaccharide (LPS) secreted by Porphyromonas gingivalis.

In the present invention, the term "halt breath" is an odor originating from the oral cavity and adjacent organs, and 85 to 90% of the bad breath originates from the oral cavity, particularly from the back of the tongue. The main components of bad breath are volatile sulfur compounds, with 90% of the total amount of volatile sulfur compounds being hydrogen sulfide made from cysteine, methyl mercaptan made from methionine, and dimethyl sulfide. These components are produced primarily by protein enzymes released by anaerobic bacteria, and the back of the tongue is the most important habitat. This part does not clean well by saliva, and has many small depressions, making it a place where bacteria can continue to 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 the development of bad breath, and non-limiting examples of bad breath-causing bacteria include Porphyromonas gingivalis , which secretes many types and large amounts of enzymes.

In the present invention, the term, " Porphyromonas gingivalis ( Porphyromonas gingivalis or P. gingivalis )" is one of several representative causative bacteria causing oral disease. The Porphyromonas ginji ballis is a type of bacterioid related bacteria and is a gram-negative bacterium and an anaerobic bacterium. The porphyromonas gingivalis occurs in the oral cavity where periodontal disease occurs, and in addition, it is also found in the upper gastrointestinal tract, respiratory tract and colon. Accordingly, oral disease can be prevented, improved or treated through antibacterial activity against Porphyromonas gingivalis.

In one embodiment of the present invention, as a result of administering the taro extract, tail fern extract or forest buckwheat extract of the present invention to Porphyromonas ginsalis, it was confirmed that it exhibits antimicrobial activity against Porphyromonas ginsalis in a concentration-dependent manner. (Fig. 4a, 4b, 4c), it was confirmed that the taro extract, tail fern extract or forest buckwheat extract of the present invention also exhibits an antibacterial lasting effect (Fig. 5a, 5b, 5c). Through this, it was confirmed that the taro extract, tail fern extract, or forest buckwheat extract is effective in preventing or treating oral diseases through antibacterial activity and antibacterial lasting activity against Porphyromonas gingivalis.

In the present invention, the term, "bone disease" means to include all diseases induced by a decrease in bone loss or bone density, the balance of osteoblast and osteoclast production and differentiation is lost. Specifically, osteoporosis, osteomalacia, osteoopenia, bone atrophy, osteoarthritis, rheumatoid arthritis, periodontal disease, osteolysis, May include fibrous dysplasia, Paget's disease, osteogenesis imperfect, hypercalcemia, and more specifically osteoporosis due to osteoclast differentiation. However, it is not limited thereto.

In the present invention, the term "osteoclast (osteoclast)" is a cell derived from a macrophage precursor (macrophage precursor), osteoclast progenitor cells are macrophage colony stimulating factor (M-CSF), NF-κB It refers to differentiation into osteoclasts by a receptor activating factor ligand (RANKL) and the like, to form multinucleated osteoclasts through fusion. Osteoclast cells bind to the bone through αvβ3 integrin, etc., create an acidic environment, and secrete various collagenase and protease to cause bone resorption. Osteoclasts do not proliferate as fully differentiated cells and cause apoptosis when the life span of about 2 weeks is reached.

In one embodiment of the present invention, after administration of the taro extract, tail fern extract or forest buckwheat extract of the present invention to mouse monocyte cells differentiated into osteoclasts, staining of tarrate-resistant acid phosphate (TRAP), an osteoclast differentiation marker, is performed. As a result, it was confirmed that osteoclast formation was inhibited by significantly inhibiting TRAP activity in a concentration-dependent manner (FIGS. 6, 7 and 8). Through this, it was confirmed that the taro extract, tail fern extract or forest buckwheat extract is effective in preventing or treating bone diseases, such as osteoporosis or oral diseases, through inhibitory activity of osteoclast formation.

In the present invention, the term, "prevention" refers to all actions to suppress or delay the development of bone disease by administration of the pharmaceutical composition according to the present invention, "treatment" suspects bone disease by administration of the pharmaceutical composition And all actions in which the symptoms of the affected individual are improved or beneficially changed.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent, which carrier may include a non-naturally occurring carrier. Specifically, carriers, excipients, and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, polycaprolactone, polylactic acid (Poly Lactic Acid), poly-L-lactic acid, Mineral oil and the like.

The pharmaceutical composition is an oral dosage form such as pills, powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injections according to conventional methods for preventing and treating oral diseases or bone diseases It can be used by formulating in the form of a solution, and in particular, when the pharmaceutical composition of the present invention is for the prevention or treatment of oral diseases, specifically, it may have a formulation of an oral spray, an oral ointment, or an oral varnish. The carrier may include various types of amorphous carriers, microspheres, nanofibers, and rosin.

In the case of formulation, it may be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc., which are usually used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in the extract and its fractions, for example, starch, calcium carbonate, It can be prepared by mixing sucrose or lactose, gelatin, and the like. In addition, lubricants such as magnesium stearate and talc may be used in addition to simple excipients.

Liquid preparations for oral administration include suspensions, intravenous solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used as diluents, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, can be included. have.

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

The content of any one or more or fractions thereof selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract contained in the pharmaceutical composition of the present invention is not particularly limited, but is 0.01 based on the total weight of the final composition It may be included in an amount of 100 to 100% by weight, 0.01 to 50% by weight, and more specifically 0.01 to 20% by weight.

The present invention provides a method for preventing or treating oral disease or bone disease, comprising administering the pharmaceutical composition to an individual.

Since the pharmaceutical composition of the present invention exhibits the prevention or treatment effect of oral disease or bone disease, the method of the present invention comprising the step of administering it to an individual may be usefully used for the prevention or treatment of oral disease or bone disease. have.

In the present invention, the term "individual" refers to all animals including humans who may or may have oral disease or bone disease, for example, monkeys, dogs, cats, rabbits, mormots, rats, mice, cows, sheep, pigs , Goat, bird, fish, etc., and specific examples may include mammals including humans, but are not limited thereto.

In the present invention, the term "administration" means to introduce the composition to the subject in any suitable way, the route of administration can be administered through a variety of routes as long as it can reach the target tissue. Specifically, the pharmaceutical composition of the present invention can be administered orally, intravenously, subcutaneously, intradermally, intranasally, intraperitoneally, intramuscularly, transdermally, etc., and suitable for topical treatment, including intralesional administration, if necessary. It can be administered by methods and routes. For example, it may be administered by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine epidural or cerebrovascular injection, but is not limited thereto.

As a treatment method of the present invention, a pharmaceutical composition containing any one or more selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract as an active ingredient may be administered in a pharmaceutically effective amount.

In the present invention, the term "pharmaceutically effective amount" means an amount sufficient to treat or prevent a disease at a reasonable benefit / risk ratio applicable to medical treatment or prevention, and the effective dose level is the severity of the disease, the activity of the drug , The patient's age, weight, health, sex, patient's sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration and the rate of excretion, the duration of treatment, the drug used in combination or coincidental with the composition of the present invention used. Factors and other factors well known in the medical field can be determined. The pharmaceutical composition of the present invention may be administered alone or in combination with a known pterygium therapeutic agent. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects.

In addition, the dosage of the pharmaceutical composition can be determined by a person skilled in the art in consideration of the purpose of use, the degree of poisoning of the disease, the age, weight, sex, history of the patient, or the type of substance used as an active ingredient. For example, the pharmaceutical composition of the present invention may be administered at 1 mg / kg to 200 mg / kg per adult, specifically 1 mg / kg to 100 mg / kg, more specifically 20 to 40 mg / kg, , The frequency of administration of the composition of the present invention is not particularly limited, but may be administered once a day or divided into doses several times. The above dosage does not limit the scope of the present invention in any way.

As another aspect, it provides a food composition for preventing or improving oral disease or bone disease, including any one or more selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract as an active ingredient.

Specifically, the food composition for preventing or improving oral disease or bone disease of the present invention may include any one or more or fractions thereof selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract.

At this time, the "taro", "tail and fern". The descriptions of "forest wheat", "extract", "fraction", "oral disease", "bone disease", and "prevention" are as described above.

In the present invention, the term, "improvement" is an oral disease that is prevented or treated by using a composition comprising at least one or a fraction thereof selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract as an active ingredient, or Refers to all actions that improve or benefit the suspected bone disease and the symptoms of the affected individual.

The term "food" in the present invention, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages , Vitamin complexes, health functional foods, and the like, all of the foods in the usual sense, and as long as it can contain the taro extract or fractions thereof of the present invention, it is not limited. It may also include forms such as pills, powders, granules, needles, tablets, capsules or liquids.

In the present invention, the term, "health functional food" refers to food manufactured and processed using ingredients or ingredients having useful functionality for the human body according to Act No. 6727 on the Health Functional Food, and 'functional' is the structure of the human body And it means to obtain a useful effect in health applications such as adjusting nutrients for function or physiological action. On the other hand, health foods mean foods that have an active health maintenance or enhancement effect compared to general foods, and health supplements mean foods for the purpose of supplementing health, and in some cases, terms of health functional foods, health foods, and health supplements Can be used interchangeably. The health functional food of the present invention can be manufactured by a method commonly used in the art. It can be manufactured in various types of formulations, and unlike general medicines, it has the advantage of not having side effects that can occur when taking medicines for a long time using food as a raw material, and can be excellent in portability.

When preparing the food composition, it may be prepared by adding raw materials and ingredients that are conventionally added in the art, and the type is not particularly limited. For example, as a conventional food, various herbal extracts, food additives, food supplements, or natural carbohydrates may be included as additional ingredients, but are not limited thereto.

As another aspect, a quasi-drug composition for preventing or improving oral disease or bone disease comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract as an active ingredient is provided.

Specifically, the quasi-drug composition for preventing or improving oral disease or bone disease of the present invention may include any one or more or fractions thereof selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract.

At this time, the description of the "taro", "tail fern", "forest buckwheat", "extract", "fraction", "oral disease", "bone disease", "prevention" and "improvement" are as described above .

In the present invention, the term "quasi-drug" is a fiber, rubber product or the like used for the purpose of treating, alleviating, treating or preventing a disease of a person or animal, has a weak effect on the human body or does not act directly on the human body, Or a non-mechanical and similar product, one of the agents used for sterilization, pesticide, and similar purposes for the prevention of infection type, and diagnoses human or animal diseases. Of items used for the purpose of treatment, alleviation, treatment, or prevention, other than devices, machines, or devices, and items used for the purpose of having a pharmacological effect on the structure and function of a person or animal, other than devices, machinery, or devices Means goods. In addition, the quasi-drug may include an external preparation for skin or a personal hygiene product.

The external preparation for skin is not particularly limited thereto, and specifically, it may be prepared and used in the form of an ointment, lotion, spray, patch, cream, powder, suspension, gel or gel. Without being limited thereto, specifically, soap, cosmetics, wipes, tissue, shampoo, skin cream, face cream, toothpaste, lipstick, perfume, makeup, foundation, ball touch, mascara, eye shadow, sunscreen lotion, hair care products, It may be an air freshener gel or a cleaning gel. In addition, another example of the quasi-drug composition of the present invention is a disinfecting cleaner, shower foam, oral brushing solution, oral spray, wipes, detergent soap, hand wash, humidifier filler, mask, ointment or filter filler.

When the taro extract of the present invention or a fraction thereof is used as a quasi-drug additive, the extract or fraction can be added as it is or used with other quasi-drugs or quasi-drug components, and can be suitably used according to a conventional method, and the active ingredient mixture amount is used It can be determined accordingly.

As another aspect, a dental composition for preventing or improving oral diseases comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract as an active ingredient is provided.

Specifically, the dental composition for preventing or improving oral disease of the present invention may include any one or more or a fraction thereof selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract.

At this time, the description of the "taro", "tail fern", "forest wheat", "extract", "fraction", "oral disease", "prevention" and "improvement" are as described above.

In the present invention, the term "dental use" may mean a use for preventing or treating diseases and damages in the teeth and its supporting tissues and oral cavity.

The dental composition can be applied to dental products such as antibacterial fluorine varnish by adding it to fluorine varnish, oral brushing solution, dental crust relieving agent, antibacterial dental adhesive, dental filling material, dental restorative material, dental It can be applied to and used in dental materials such as root canal fillers, dental root canal sealers, sonar fissure sealants, dental coatings, and dental cement.

In addition, the dental composition can be used for all articles used for oral health. Examples of the article include dental products such as dental floss, toothbrush, interdental toothbrush, electric toothbrush, tongue cleaner, mouthwash, toothbrush sterilizer, dental devices such as braces, implants, or all dental tools and instruments used in dental care. However, it is not limited thereto.

Specifically, the article is immersed in the dental composition, the dental composition is applied or sprayed on the surface of the article, the surface of the article is washed several times with the dental composition, or the dental composition seeps Any towel or the like can be used for the article by a method such as wiping the surface of the article, and the method is not particularly limited as long as the article and the dental composition can be contacted for a predetermined time. In addition, the time for performing the method is not limited as long as the dental composition can react sufficiently with the surface of the article, and those skilled in the art can appropriately select it.

Hereinafter, the present invention will be described in more detail by the following examples. However, the following examples are only for illustrating the present invention and the scope of the present invention is not limited to them.

Example 1. Preparation of each plant extract

Through the cooperation of domestic plant classifiers, cultivation experts, and herbal medicine discrimination experts, native taro, tail fern and forest wheat were collected in Korea, and dried outposts were put in 99.9% methanol and extracted at 45 ° C for 3 days. After 15 minutes of sonication at the time of extraction, a 2 hour suspension was performed 10 times per day. Subsequently, it is filtered using a non-fluorescent cotton filter, concentrated using a rotary evaporator (N-1000SWD; EYELA) at 45 ° C, and 24 using a module spin 40 (Biotron co.). Dry for an hour. Thereafter, the extract was removed in the form of 20 mg / tube, and then stored in a low temperature room at -4 ° C. The above procedure was performed at the Korea Plant Extract Bank, and a sample of 20 mg / tube was dissolved in DMSO for use in cell experiments to a concentration of 50 mg / ml, which was used by diluting it with a cell culture medium.

Example 2. Anti-inflammatory effect of each extract

RAW264.7 cells were purchased from ATCC (Rockville, MD), 10% fetal bovine serum (FBS, HyClone Logaa, UT), streptomycin (100 μg / mL), and penicillin (100 U / mL) The added growth medium (CM, DMEM) was cultured in humidified air at 37 ° C and 5% CO ₂ .

Example 2-1. Cytotoxicity evaluation

4Х10 ³ RAW264.7 cells were plated on GM's 96-well plate. After incubation for 24 hours, in the presence of lipopolysaccharide (LPS, 1 μg / mL) in cells, the taro extract, tail fern extract and forest buckwheat extract prepared in Example 1 for 1 day were each concentration (0, 3, 10). And 30 μg / mL). Cell viability (%) was measured three times using Cell Counting Kit-8 (CCK-8, Dojindo, Rockville, ML, USA) according to the manufacturer's manual. Absorbance was measured with a microplate reader (VERSA max ^™ , Molecular Devices, Sunnyvale, CA) and converted to cell number using a standard curve.

As a result, as shown in FIGS. 1A, 1B, and 1C, when the taro extract, tail fern extract, and forest buckwheat extract were respectively treated, it was confirmed that cell viability was not exhibited by showing cell viability close to 100% at all concentrations. These results show that the antimicrobial effect or osteoclast production inhibitory effect to be confirmed later is not due to the toxic effect on cells, but the antibacterial effect or osteoclast production inhibitory effect by taro extract, tail fern extract or forest buckwheat extract.

Example 2-2. Nitric oxide production inhibitory activity

The cells were streaked with 1Х10 ⁶ cells in a 60 mm dish, and each of the taro extract, tail fern extract and forest buckwheat extract prepared in Example 1 for 2 hours in concentration (0, 3, 10 and 30 μg / mL) Treatment. Then, LPS (1 μg / mL) was added. After incubation for 24 hours, nitrogen oxide (NO) levels were analyzed using a grease reaction. Cell culture medium (100 μL) was mixed with grease reagent and incubated for 10 minutes at room temperature. Absorbance was measured with a microplate reader at 540 nm. As a blank in all experiments, fresh culture medium was used. Nitrite (NO ₂ ) was measured by a standard curve using sodium nitrite (NaNO ₂ ).

As a result, as shown in FIGS. 2A, 2B, and 2C, when only LPS was added, the level of nitrogen oxide was significantly increased compared to the control group, but only LPS was added when LPS was added after treatment with taro extract, tail fern extract or forest buckwheat extract It was confirmed that the nitrogen oxide level was significantly reduced than in one case. These results show that the taro extract, tail fern extract or forest buckwheat extract exhibits anti-inflammatory effects by inhibiting the production of nitric oxide, an inflammatory mediator in cells.

Example 3. Antibacterial effect of each extract

Example 3-1. Antibacterial activity of each extract

To evaluate the oral antimicrobial activity of taro extract, tail fern extract or forest buckwheat extract, a bacterium Porphyromonas gingivalis ( P. gingivalis , ATCC33277 ) was used. BHI broth containing hemin and menadione was used as a medium to activate Porphyromonas gingivalis bacteria, and after incubation for 72 hours in a 37 ° C CO ₂ incubator, each well of a 96-well plate ( into the well) 90 μL each, 10 μL each of the taro extract, tail fern extract and the forest buckwheat extract prepared in Example 1 were added. As a control, 100 μL of only bacteria was used, and 100 μL of BHI broth was additionally added to each well, resulting in a final concentration of 125 μg / mL, 250 μg / mL, and 500 μl of taro extract, tail fern extract, or forest wheat extract. It was made to be μg / mL. Thereafter, the sample was cultured for 72 hours. Absorbance was measured with an ELISA kit (Spectra MAX250, Molecular Devices Co.) at 600 nm to determine the concentration at which growth of bacteria was inhibited.

As a result, as shown in Figures 4a, 4b, 4c, when processing the taro extract, tail fern extract or forest buckwheat extract, it was confirmed that the concentration-dependent growth of Porphyromonas gingivalis. Specifically, the growth of Porphyromonas gingivalis is about 20% inhibited when 500% of the concentration of 125 μg / ml and 250 μg / ml of taro extract, tail fern extract or forest buckwheat extract is treated, respectively, of 500 μg / ml When each extract was treated, it was confirmed that the growth of Porphyromonas gingivalis was inhibited by about 40%.

Example 3-2. Antibacterial lasting activity of each extract

Evaluation of the antimicrobial sustained activity of the taro extract, tail fern extract or forest buckwheat extract against Porphyromonas gingivalis was performed as follows. The film (OHP film) in the form of a disk with a diameter of 5 mm was put in sterile packaging and sterilized with EO gas, and then a sample was prepared as shown in Table 1, and evenly applied to the entire surface of the film with the same thickness, evenly filling the entire surface. At this time, three films were made per sample, placed in a petri dish with a diameter of 35 mm, and dried in a clean bench for 30 minutes while UV was turned on. After adding 2000 μL of sterilized distilled water to a petri dish with a film coated with a sample, the coated surface was directed downward so that the sample could be eluted in water. After storing at 80 rpm in a shaking water bath at 37 ° C. for 3 days, the film was taken out from the petri dish and the antibacterial activity was evaluated using an agar diffusion test.

Sample 1 (control)	Film itself
Sample 2	Fluorine varnish KR610
Sample 3	Taro 50 mg / ml (50 μL) + Fluorine varnish KR610 (50 μL)
Sample 4	Tail fern 50 mg / ml (50 μL) + Fluorine varnish KR610 (50 μL)
Sample 5	Forest wheat 50 mg / ml (50 μL) + Fluorine varnish KR610 (50 μL)

The agar diffusion method was performed as follows. Porphyromonas gingivalis was incubated in a BHI broth medium in a 37 ° C. CO ₂ incubator for 72 hours, and then inoculated onto agar medium coated with a base agar.

After the top agar was hardened, the film stored at 80 rpm in a 37 ° C. shaking bath for 3 days was placed on an agar medium. At this time, the surface of the film coated with the sample was allowed to be bonded to the agar medium downward. Subsequently, after incubation in a CO ₂ incubator at 37 ° C. for 72 hours, the diameter of the Porphyromonas gingivalis inhibitor band formed around the OHP film was measured in a right angle direction to determine the average as an inhibition zone (mm).

As a result, as shown in FIGS. 5A, 5B, and 5C, the fluoride varnish treatment group showed a significant antibacterial effect compared to the control group, and the fluoride varnish + taro treatment group, the fluoride varnish + tail fern treatment group, and the fluoride varnish + forest remodeling. It was confirmed that all the treatment groups exhibited a significant antibacterial lasting effect compared to the fluoride varnish treatment group.

Through the above results, the taro extract, tail fern extract or forest buckthorn extract of the present invention exhibits excellent antibacterial and antibacterial lasting effects against strains causing oral diseases, so the composition comprising any one or more of the extracts may be used for oral diseases. It was found that it can be effectively used for treatment.

Compared to the control group, the bovine varnish treatment group showed an increase of about 60%, and the treatment group of the combination of the fluoride varnish and each extract increased about 150% compared to the control group, and about 50% compared to the control group. It was confirmed to increase.

Example 4. Inhibitory effect of osteoclast differentiation of each extract

A 5 week old male ICR mouse was purchased and the femur and tibia were taken and washed with α-MEM containing antibiotics (100 units / mL penicillin and 100 μg / mL streptomycin) to bone marrow cells ). The bone marrow cells were cultured for 1 day in α-MEM containing 10% fetal bovine serum (FBS) and macrophage colony-stimulating factor (M-CSF) 10 ng / mL. Non-adherent bone marrow cells were dispensed into Petri dishes and cultured for 3 days in the presence of M-CSF (30 ng / mL). After washing non-adherent cells, adherent cells were used as bone marrow-derived macrophage (BMM). In order to differentiate BMM cells into osteoclasts, BMM cells (1 Х 104 cells / well) were applied to a 96-well plate in the presence of RANKL (10 ng / mL) and M-CSF (30 ng / mL), and Examples Each extract prepared in 1 was treated by concentration (0, 3, 10 and 30 μg / mL), and the final concentrations were 125, 250 and 500 μg / ml, and cultured for 4 days. As a negative control, cells treated with RANKL only were used. After incubation for 4 days, multinucleated osteoclasts were fixed with 3.7% formalin for 10 minutes and treated with 0.1% Triton X-100 for 10 minutes to obtain permeability. The osteoclasts were stained red by treatment with TRAP solution (Sigma Aldrich, USA). Thereafter, in order to measure the activity of osteoclast differentiation marker, tartrate-resistant acid phosphatase (TRAP), 100 μl of TRAP buffer solution (100 mM sodium citrate pH 5.0, 50 mM sodium tartrate, 3 mM p-nitrophenyl phosphate) was added and 37 After reacting at ° C for 5 minutes, the reaction was terminated by adding 100 μl of 0.1 N NaOH. Thereafter, the absorbance at 405 nm was measured to measure TRAP activity.

As a result, as shown in Figures 6, 7 and 8, when treated with taro extract, tail fern extract or forest buckwheat extract respectively to RANKL-derived osteoclasts stained red while inhibiting TRAP activity in a concentration-dependent manner It was confirmed that the number of osteoclasts was significantly reduced.

Specifically, when treated with 30 μg / mL taro extract showed a TRAP activity inhibition rate of about 50%, and when treated with a tail fern extract 30 μg / mL, it showed a TRAP activity inhibition rate of about 25%, forest wheat extract 30 μg / It was confirmed that the TRAP activity of about 14% was suppressed when the mL was treated.

As described above, the taro extract, tail fern extract or forest buckwheat extract of the present invention exhibits excellent osteoclast differentiation and production inhibitory effect through the above results, so the composition containing any one or more of the extracts is used for the treatment of oral diseases and bone diseases. It was found that it can be effectively used.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be implemented in other specific forms without changing its technical spirit or essential characteristics. In this regard, the embodiments described above should be understood as illustrative in all respects and not restrictive. The scope of the present invention should be construed as including all changes or modifications derived from the meaning and scope of the following claims rather than the above detailed description and equivalent concepts thereof.

Claims (11)

1. A pharmaceutical composition for the prevention or treatment of oral disease or bone disease, comprising any one or more selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract.
2. The pharmaceutical composition of claim 1, wherein the extract is extracted with a solvent selected from the group consisting of water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.
3. The pharmaceutical composition of claim 1, wherein the oral disease is periodontal disease or bad breath.
4. The pharmaceutical composition according to claim 3, wherein the periodontal disease is periodontitis, gingivitis or a combination thereof.
5. The method of claim 1, wherein the bone disease is osteoporosis (osteoporosis), osteomalacia, osteopenia, bone atrophy, osteoarthritis, rheumatoid arthritis, periodontal disease. , Osteolysis, fibrous dysplasia, Paget's disease, osteogenesis imperfect, hypercalcemia, any one or more selected from the group consisting of, pharmaceutical composition .
6. The pharmaceutical composition of claim 1, wherein the bone disease is osteoporosis.
7. The pharmaceutical composition of claim 1, wherein the prevention or treatment of the oral disease is achieved through the inhibition of Porphyromonas gingivalis activity.
8. The pharmaceutical composition of claim 1, wherein the prevention or treatment of the oral disease or bone disease is achieved through suppression of osteoclast differentiation.
9. A food composition for preventing or improving oral disease or bone disease, comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract.
10. A quasi-drug composition for the prevention or improvement of oral diseases or bone diseases, comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract.
11. Dental composition for the prevention or improvement of oral diseases comprising at least one selected from the group consisting of taro extract, tail fern extract and forest buckwheat extract.

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