KR102334997B1 - Compositions for preventing or treating oral disease or bone disease comprising extracts of Sagittaria genus - Google Patents

Abstract

The present invention relates to a composition for preventing or treating oral diseases or bone diseases comprising a plant extract of the genus fluff, and specifically, a pharmaceutical for preventing or treating oral diseases or bone diseases comprising a plant extract of the genus fluff as an active ingredient; It relates to a composition, a food composition, a quasi-drug composition, and a dental composition.
The fluff extract of the present invention is derived from a natural product and has low toxicity, and significantly inhibits the activity of Porphyromonas gingivalis, an oral disease-causing bacterium, and significantly inhibits the formation or differentiation of osteoclasts, , it can be usefully used for the prevention or treatment of oral diseases or bone diseases by promoting the differentiation of osteoblasts.

Description

A composition for preventing or treating oral disease or bone disease comprising a plant extract of the genus fluff as an active ingredient {Compositions for preventing or treating oral disease or bone disease comprising extracts of Sagittaria genus}

The present invention relates to a composition for preventing or treating oral diseases or bone diseases comprising a plant extract of the genus fluff, specifically, a pharmaceutical composition for preventing or treating oral diseases or bone diseases comprising an extract of a plant of the genus fluff as an active ingredient; It relates to a composition, a food composition, a quasi-drug composition, and a dental composition.

Osteoclasts cause the destruction and resorption of abnormal bone tissue due to imbalance with osteoblasts in the bone. Causes such as fibrous dysplasia, in which bone tissue is replaced by fibrous connective tissue and immature bone trabeculae, periodontal disease in which alveolar bone is lost, and rheumatoid arthritis that causes joint destruction and deformation is known to be As such, there is a close relationship between the differentiation of osteoclasts and periodontal disease and bone disease.

In the case of Porphyromonas gingivalis, it is known that lipopolysaccharide (LPS) or cilia-mediated host-mediated inflammatory response inhibits the differentiation of osteoblasts, thereby reducing osteoogenesis (Wilson, 1998; Kadono et al., 1999), the LPS is a pathogenic component present in the bacterial outer membrane and is a well-known stimulator of inflammatory cytokines or eicosanoid synthesis. It is known as a major structure that secretes nitrogen and destroys periodontal tissue. In addition, the differentiation of osteoclasts is directly promoted and activated by LPS itself, which can ultimately induce bone loss (Koide et al., 2000; Taubman et al., 2005; Islam et al., 2007; Hou et al., 2013; Teramachi et al. 2017). As such, there are a number of data that have studied the effects of periodontal disease-inducing strains on the differentiation of osteoblasts or osteoclasts, and there is a close relationship between periodontal disease and bone disease, as there are therapeutic agents that can treat osteoarthritis and periodontal disease together. this exists

Osteoblasts originate from mesenchymal stem cells and are formed. Mineralization including calcium formed by the differentiation of osteoblasts can maintain bone strength as well as calcium and hormones throughout the body. It also plays a very important role in metabolic homeostasis.

The compound that promotes osteoblast activity can be used not only for the prevention or treatment of diseases and symptoms such as osteoporosis and fracture, but also for the treatment of periodontal disease for the purpose of bone strengthening (Zhang et al., Shanghai Kou Qiang). Yi Xue 7(2), pp99-103, 1998; Cahill et al, Biol Blood Marrow Transplant 10(10), pp709-717, 2004), also studies suggesting that it can be usefully used in the treatment of bone growth disorders resulting from burns (Klein et al., Osteoporos Int. 16(6), pp631-635, 2005).

As such, the differentiation of osteoblasts and periodontal disease and bone disease are closely related.

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 cause the appearance of resistant bacteria in the oral cavity and superinfection along with systemic side effects on our body, long-term use is difficult, so it can be used only as a therapeutic agent. In addition, raw Guinarine has the disadvantage of being unclear in its effect on bacteria in the oral cavity, its therapeutic effect on periodontal disease, as well as its high price. It exhibits only a temporary effect, but there is a disadvantage that it may also have a detrimental effect on the tissue when used for a long period of time. In addition, piroxide, which is recently added for whitening effect, is toxic to bacteria, but at the same time, it is also toxic to human tissues, so there is a problem in safety and sometimes bacteria resistant to pyroxide appear in bacteria. In addition, chlorhexidine is known to be the best known agent for the prevention and treatment of periodontal disease as well as the inhibition of plaque formation, but it causes tissue irritation, tissue coloration and degeneration, and has a particularly irritating taste and strong odor. There are disadvantages in that it cannot be used for a long period of time for the purpose of treatment or, in particular, prevention, such as, can cause mycosis mycosis, and has carcinogenic properties, limiting its use in the case of pregnant women. Therefore, there is a need to develop a drug capable of effectively inhibiting the growth of strains causing oral diseases without fear of side effects as described above.

Currently, bisphosphonate-based therapeutic agents are widely used to treat bone damage caused by osteoclasts, such as osteoporosis. However, the number of cases of various side effects such as osteonecrosis, severe atrial fibrillation, bone or joint incapacity, and musculoskeletal pain in patients taking bisphosphonate drugs is increasing every year (Br J Cancer 98:1736). -1740 (2008)). Therefore, there is a need to develop a drug that can compensate for the disadvantages of the existing bisphosphonate-based drugs, have low toxicity, and can effectively inhibit the formation and differentiation of osteoclasts.

Under this background, the present inventors have low toxicity derived from natural products, inhibit the activity of oral disease-causing bacteria, inhibit the formation or differentiation of osteoclasts, and to find a compound that can promote the formation or differentiation of osteoblasts As a result of diligent research, the plant extract of the fluff remarkably inhibits the activity of Porphyromonas gingivalis, an oral disease-causing bacterium, inhibits the formation or differentiation of osteoclasts, and inhibits the formation or differentiation of osteoblasts. By confirming that it promotes, the present invention has been completed by confirming that it can be usefully used for the prevention or treatment of oral diseases or bone diseases.

One object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of oral diseases or bone diseases comprising a plant extract of the genus fluff as an active ingredient.

Another object of the present invention is to provide a food composition for the prevention or improvement of oral disease or bone disease comprising a plant extract of the genus fluff 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 a plant extract of the genus fluff as an active ingredient.

Another object of the present invention is to provide a dental composition for preventing or improving oral diseases comprising a plant extract of the genus fluff as an active ingredient.

Another object of the present invention is to provide an antibacterial composition for Porphyromonas gingivalis, comprising a plant extract of the genus fluff as an active ingredient.

One embodiment of the present invention for achieving the above object provides a pharmaceutical composition for the prevention or treatment of oral disease or bone disease comprising a plant extract of the genus fluff as an active ingredient.

Specifically, the pharmaceutical composition for preventing or treating oral disease or bone disease of the present invention may include a plant extract of the genus fluff or a fraction thereof.

As used herein, the term “plants of the genus fluff” may include, but is not limited to, fluff, bovine herbaceous plant, and ragweed.

The fluff (Sagittaria aginashi) is a perennial herb of the family Taxaceae, a monocotyledonous plant, and it is known that the leaves are clustered in the rhizome and have long petioles in the shape of arrowheads.

The small gwi namul (Sagittaria trifolia var. edulis) is a perennial grass of the monocotyledonous plant family of the sagittal family, also called jago (慈姑). It grows in water and is known to have underground stems extending to the side and tubers at the end.

Said grass (Sagittaria sagittifolia L.) is a perennial grass of the monocotyledonous progenitor family Taxaceae, which grows in wetlands or shallow water, and is known to have a small cortex at the end of a branch extending sideways.

The present inventors have conducted various studies on substances that have excellent preventive or therapeutic effects for oral diseases or bone diseases from natural resources present in nature. gingivalis) and inhibiting the formation or differentiation of osteoclasts, it was confirmed that it can be usefully used for the prevention or treatment of oral diseases or bone diseases. Through this, it was found that the plant extract of the genus fluff can be used as a pharmaceutical composition for the prevention or treatment of oral diseases or bone diseases. A composition comprising a plant extract of the genus fluff having a preventive or therapeutic effect for oral disease or bone disease has not been known so far, and it can be said that it is very significant in that it was first developed by the present inventors.

For the purpose of the present invention, plants of the genus fluff are not limited to the type, and cultivated ones may be used, commercially purchased ones, or used without limitation on the source.

As used herein, the term "extract" refers to the extraction treatment of the plants of the genus fluff, but is not limited thereto, but the extract obtained by the extraction treatment, the diluted or concentrated solution of the extract, the dried product obtained by drying the extract, and their It may include extracts of all formulations that can be formed using the extract itself and the extract, such as a crude product, a purified product, or a mixture thereof. In addition, it can be extracted from various organs of natural, hybrid, and mutated plants of the genus plant, for example, roots, rhizomes, above-ground parts, stems, leaves, fruits, and the like.

The plant extract of the genus fluff 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 methods such as immersion extraction, hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction may be used.

The fluff plant extract can be extracted with water, an alcohol having 1 (C₁) to 4 (C₄) carbon atoms, or a mixed solvent thereof, but is not limited thereto, and the degree of extraction and loss of the active ingredient according to the organic solvent to be extracted Since the degree may be different, it is possible to select and use an appropriate organic solvent. 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 suspended solid particles. Specifically, it is possible to filter particles using nylon or the like, or to use ultrafiltration, cryofiltration, centrifugation, etc., but is not limited thereto.

Methods such as reduced pressure concentration, reverse osmosis concentration, etc. may be used for concentration of the extract.

For the drying step after concentration, freeze drying, vacuum drying, hot air drying, spray drying, reduced pressure drying, foam drying, high frequency drying, or infrared drying may be used. In some cases, it may further include a process of pulverizing the final dried extract.

As used herein, the term "fraction" refers to a result obtained by performing fractionation in order to separate a specific component or a specific component group from a mixture including various components.

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

In the present invention, the type of the fractionation 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 fractionation solvent include a polar solvent such as water, distilled water, alcohol; Non-polar solvents, such as hexane, ethyl acetate, chloroform, and dichloromethane, etc. are mentioned. These may be used alone or in combination of two or more. In the case of using an alcohol in the fractionation solvent, an alcohol having 1 (C₁) to 4 (C₄) carbon atoms may be preferably used.

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

In the present invention, “oral disease” refers to various diseases occurring in the oral region, and the oral region refers to a space in the mouth connected from the front lip to the pharynx in the rear palate. In the present invention, the oral disease is a concept that includes any disease occurring in the oral cavity, regardless of the pathology, and non-limiting examples of the oral disease include dental caries, periodontal disease (periodontitis or gingivitis), toothache, sore throat, bad breath. and the like, preferably periodontitis or gingivitis.

As used herein, the term "periodontal disease" is often referred to as gingivitis, and is divided into gingivitis and periodontitis according to the severity of the disease. It is a relatively light and fast recovery form of oral disease limited only to the gums, that is, soft tissues, and is called gingivitis.

There is a V-shaped gap between the gingiva (gum) and the teeth. Periodontal disease is when bacteria attack the subgingival part of the gingival sulcus and damage the periodontal ligament and adjacent tissues. As inflammation progresses and more tissue is damaged, the groove develops into a periodontal pocket. As the periodontal pocket deepens, the periodontal ligament becomes inflamed, and osteoclast differentiation is promoted by lipopolysaccharide (LPS) secreted by Porphyromonas gingivalis, leading to loss of alveolar bone.

As used herein, the term "Porphyromonas gingivalis or P. gingivalis" is one of several representative causative bacteria that cause oral diseases. The Porphyromonas gingivalis is a kind of Bacteroid-like bacteria and is a Gram-negative bacteria and anaerobic bacteria. The Porphyromonas gingivalis occurs in the oral cavity where periodontal disease has occurred, and is also found in the upper gastrointestinal tract, respiratory tract and colon. Accordingly, oral diseases 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 plant extract of the genus fluff of the present invention to Porphyromonas gingivalis, it was confirmed that antibacterial activity against Porphyromonas gingivalis was shown (FIG. 1). Through this, it was confirmed that the plant extract of the genus fluff was effective in preventing or treating oral diseases through antibacterial activity against Porphyromonas gingivalis.

As used herein, the term "bone disease" means to include all diseases induced by bone loss or a decrease in bone density, which appear due to a loss of balance between the generation and differentiation of osteoblasts and osteoclasts. Specifically, osteoporosis, osteomalacia, osteopenia, bone atrophy, osteoarthritis, rheumatoid arthritis, periodontal disease, osteolysis, It may include fibrous dysplasia, Paget's disease, osteogenic imperfect, hypercalcemia, etc., and more specifically, osteoporosis due to osteoclast differentiation. However, the present invention is not limited thereto.

As used herein, the term "osteoclast" is a cell derived from a macrophage precursor, and the osteoclast precursor cells are macrophage colony stimulating factor (M-CSF), NF-κB It is differentiated into osteoclasts by receptor activator ligand (RANKL), etc., and means to form multinucleated osteoclasts through fusion. Osteoclasts bind to bone through αvβ3 integrin, etc. and create an acidic environment, while secreting various collagenases and proteases to cause bone resorption. Osteoclasts do not proliferate into fully differentiated cells and cause apoptosis at the end of their life span of about 2 weeks.

More specifically, as for osteoclasts, RAW264.7 monocytes from mice are differentiated into multinucleated osteoclasts by RANKL (receptor activator of nuclear factor ĸB (RANK) ligand). In this differentiation process, RANKL outside the cell binds to RANK and promotes the activity of mitogen-activated protein kinase (MAPK). It is possible by increasing the expression of (tartrate-resistant acid phosphatase), MMP-9 (matrix metalloproteinase-9), and c-Src tyrosine kinase, etc. acts as an absorber. In addition, when RANKL binds to RANK, it promotes the activity of tumor necrosis factor receptor-associated factor 6 (TRAF6), thereby promoting the activity of MARK, or transcription factors such as NF-ĸB, AP-1, and 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, it is known that blocking the signaling pathway activated by RANKL is recognized as one of the therapeutic approaches in the treatment of diseases related to osteoclast differentiation.

As used herein, the term “osteoblast” refers to a cell that is generated by differentiation from mesenchymal stem cells, and plays a role in increasing bone density by making bone mass.

More specifically, representative factors in the formation of calcium by differentiation of osteoblasts include bone morphogenetic protein-2 (BMP-2) and runt-related transcription factor2 (RUNX2). BMP-2 not only regulates its own gene expression by promoting the activity of RUNX2, but also increases the expression of various genes involved in osteoblast differentiation. During osteoblast differentiation, alkaline phosphatase (ALP) is synthesized in the initial differentiation stage, and then osteocalcin (OCN), osteopontin (OPN), type I collagen (COLLI) involved in mineralization. ), etc., are known to form bone by differentiation of osteoblasts.

In one embodiment of the present invention, after administration of the fluff extract of the present invention to mouse mononuclear cells differentiated into osteoclasts, TRAP (tarratresistant acid phosphate), a marker for osteoclast differentiation, was stained. As a result, concentration-dependent TRAP activity was significantly increased. It was confirmed that (FIG. 2), differentiation into multinucleated osteoclasts was inhibited (FIG. 3), the activity of ALP, an osteoblast differentiation marker, was promoted (FIG. 4), and differentiation of osteoblasts was promoted (FIG. 5). Through this, it was confirmed that the fluff extract is effective in preventing or treating bone diseases, such as osteoporosis or oral diseases, through the activity of inhibiting osteoclast formation and promoting osteoblast formation.

As used herein, the term "prevention" refers to any act of inhibiting or delaying the onset of bone disease by administration of the pharmaceutical composition according to the present invention, and "treatment" is suspected of bone disease by administration of the pharmaceutical composition. And it means any action in which the symptoms of the affected individual are improved or changed to a beneficial effect.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent, which 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 gum, alginate, gelatin, calcium phosphate, calcium Silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, polycaprolactone, polylactic acid Lactic Acid), poly-L-lactic acid, and mineral oil.

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

In the case of formulation, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used.

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

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

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

The content of the fluff plant extract or a fraction thereof contained in the pharmaceutical composition of the present invention is not particularly limited thereto, but 0.01 to 100% by weight, 0.01 to 50% by weight, more specifically 0.01 to 20% by weight based on the total weight of the final composition It may be included in a content of wt%.

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

Since the pharmaceutical composition of the present invention exhibits a preventive or therapeutic effect on oral disease or bone disease, the method of the present invention comprising the step of administering it to an individual can be usefully utilized for the prevention or treatment of oral disease or bone disease. have.

As used herein, the term "individual" refers to any animal, including humans, that has or can develop oral disease or bone disease, for example, monkey, dog, cat, rabbit, guinea pig, rat, mouse, cow, sheep, pig. , goats, birds, fish, and the like, and as a specific example, may be mammals including humans, but is not limited thereto.

As used herein, the term "administration" means introducing the composition to an individual by any suitable method, and the administration route may be administered through various routes as long as it can reach a target tissue. Specifically, the pharmaceutical composition of the present invention may be administered orally, intravenously, subcutaneously, intradermally, intranasally, intraperitoneally, intramuscularly, transdermally, etc., and if necessary for local treatment, suitable including intralesional administration It may be administered by any method or route. For example, it may be administered by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebrovascular injection, but is not limited thereto.

In the treatment method of the present invention, a pharmaceutical composition comprising an extract in fluff may be administered in a pharmaceutically effective amount.

As used herein, 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 depends on the severity of the disease, the activity of the drug. , patient's age, weight, health, sex, patient's sensitivity to drug, administration time of the present composition used, administration route and excretion rate, treatment period, including drugs used in combination with or concurrently with the composition of the present invention factors and other factors well known in the medical art. The pharmaceutical composition of the present invention may be administered alone or in combination with a known therapeutic agent for pterygium. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects.

In addition, the dosage of the pharmaceutical composition can be determined by those skilled in the art in consideration of the purpose of use, the degree of addiction of the disease, the age, weight, sex, history, or the type of material 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, specifically 1 mg/kg to 100 mg/kg, more specifically 20 to 40 mg/kg per adult. , The frequency of administration of the composition of the present invention is not particularly limited thereto, but may be administered once a day or administered several times by dividing the dose. 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 comprising an extract in fluff as an active ingredient.

Specifically, the food composition for preventing or improving oral disease or bone disease of the present invention may include an extract in fluff or a fraction thereof.

In this case, the descriptions of the “plants in the genus fluff”, “extracts”, “fractions”, “oral diseases”, “bone diseases” and “prevention” are the same as those described above.

As used herein, the term “improvement” refers to any action that improves or benefits the symptoms of suspected or affected individuals of oral disease or bone disease that are prevented or treated using a composition containing an extract or a fraction thereof as an active ingredient in fluff .

As used herein, the term "food" refers to meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages , vitamin complexes, health functional foods, etc. include all foods in the conventional sense, and are not limited thereto, as long as they can contain the forest foreskin extract or a fraction thereof of the present invention. In addition, it may include the form of pills, powders, granules, needles, tablets, capsules or liquids.

As used herein, the term “health functional food” refers to food manufactured and processed using raw materials or ingredients useful for the human body in accordance with Act No. 6727 of the Health Functional Food Act, and 'functionality' refers to the structure of the human body. And it means to obtain a useful effect for health use, such as regulating nutrients for function or physiological action. On the other hand, health food means food that has an active health maintenance or promotion effect compared to general food, and health supplement means food for the purpose of health supplementation. In some cases, the terms health functional food, health food and health supplement food can be mixed. The health functional food of the present invention can be prepared by a method commonly used in the art. It can be manufactured in various types of dosage forms, and unlike general drugs, it has the advantage of not having side effects that may occur during long-term administration of the drug using food as a raw material, and can be excellent in portability.

The food composition may be prepared by adding raw materials and components commonly added in the art, and the type thereof is not particularly limited. For example, it may include, as an additional component, various herbal extracts, food-logically acceptable food supplements or natural carbohydrates, such as conventional food, but is not limited thereto.

As another aspect, it provides a quasi-drug composition for preventing or improving oral disease or bone disease, comprising an extract in fluff as an active ingredient.

Specifically, the quasi-drug composition for preventing or improving oral disease or bone disease of the present invention may include an extract in fluff or a fraction thereof.

In this case, the descriptions of the “fluff”, “extract”, “fraction”, “oral disease”, “bone disease”, “prevention” and “improvement” are the same as described above.

As used herein, the term “quasi-drugs” refers to fibers, rubber products, or similar products used for the purpose of treating, alleviating, treating or preventing diseases of humans or animals, and has a weak action on the human body or does not act directly on the human body. Or non-machines and similar ones, and products corresponding to one of the preparations used for sterilization, insecticide, and similar purposes for the prevention of infection type, for diagnosing diseases of humans or animals. Items used for the purpose of treatment, alleviation, treatment or prevention, which are not instruments, machines, or devices, and items used for the purpose of pharmacologically affecting the structure and function of humans or animals, other than those that are not instruments, machines or devices means goods. In addition, the quasi-drugs may include external preparations for skin or personal hygiene products.

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, and the personal hygiene product is particularly Specifically, but not limited to, soap, cosmetics, wet tissue, tissue paper, shampoo, skin cream, face cream, toothpaste, lipstick, perfume, makeup, foundation, cheek, 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 disinfectant cleaner, shower foam, mouthwash, oral spray, wet tissue, detergent soap, hand wash, humidifier filler, mask, ointment or filter filler.

In the case of using the extract or a fraction thereof of the present invention as a quasi-drug additive, the extract or fraction can be added as it is or used together with other quasi-drugs or quasi-drug ingredients, and can be used appropriately according to a conventional method, and the active ingredient mixture is used It may be appropriately determined depending on the purpose.

As another aspect, it provides a dental composition for preventing or improving oral diseases comprising an extract in fluff as an active ingredient.

Specifically, the dental composition for preventing or improving oral disease of the present invention may include an extract in fluff or a fraction thereof.

At this time, the descriptions of the “plants in the genus fluff”, “extracts”, “fractions”, “oral diseases”, “prevention” and “improvement” are the same as described above.

As used herein, the term "dental use" may refer to a use for preventing or treating diseases or damage in teeth and their supporting tissues and oral cavity.

The dental composition can be added to fluorine varnish and applied to dental products such as antibacterial fluorine varnish, oral gargling solution, dental hypersensitivity treatment, antibacterial dental adhesive, dental filling material, dental restoration material, dental use. It can be applied to and used in dental materials such as root canal fillers, dental root canal sealers, fissure sealants, dental coatings, and dental cements.

In addition, the dental composition may be used in all articles used for oral health. Examples of the article include dental appliances such as dental floss, toothbrushes, interdental toothbrushes, electric toothbrushes, tongue cleaners, mouthwashes, toothbrush sterilizers, etc., dental devices such as braces and implants, or all dental tools and devices used in dental care. , but 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 is permeated It can be used on the article by a method such as wiping the surface of the article with a towel or the like, and is not particularly limited as long as the article and the dental composition can be in contact for a predetermined period of time. In addition, the time for performing the method is not limited as long as the dental composition can sufficiently react with the surface of the article, and a person skilled in the art can appropriately select it.

As another aspect, it provides an antibacterial composition for Porphyromonas gingivalis, comprising a plant extract of the genus fluff as an active ingredient.

At this time, the description of the terms “plants of the genus fluff” and “extract” is the same as described above.

As used herein, the term “antibacterial” may refer to a use for inhibiting the growth of the microorganism Porphyromonas gingivalis.

The fluff extract of the present invention is derived from a natural product and has low toxicity, and significantly inhibits the activity of Porphyromonas gingivalis, an oral disease-causing bacterium, and significantly inhibits the formation or differentiation of osteoclasts, , it can be usefully used for the prevention or treatment of oral diseases or bone diseases by promoting the differentiation of osteoblasts.

1 is a graph showing the growth inhibitory effect of Porphyromonas gingivalis according to the treatment concentration of a plant extract of the genus fluff.
2 is a graph showing the TRAP activity inhibitory effect according to the treatment concentration of the fluff extract.
3 is a diagram showing the effect of inhibiting differentiation into osteoclasts according to the treatment concentration of the fluff extract.
4 is a graph showing the ALP activity promoting effect according to the treatment concentration of the fluff extract.
5 is a diagram showing the effect of promoting differentiation into osteoblasts according to the fluff extract treatment.

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

Example 1: Preparation of a plant extract of the genus Fluff

In cooperation with domestic botanical taxonomists, cultivation specialists, and herbal medicine differentiation experts, the fluff, sagebrush, and wild grass native to Korea were collected, dried, put in 99.9% methanol, and extracted at 45°C for 3 days. After 15 minutes of sonication during extraction, a 24-hour stop process was performed 10 times/day. Thereafter, filtration was performed using a non-fluorescent cotton filter, and concentration was performed using a rotary evaporator (N-1000SWD; EYELA) at 45° C., and 24 using a module spin 40 (Biotron co.). dried for an hour. Then, the extract was stored in a low temperature room at -4 ℃ after removing the solvent in the form of 20 mg/tube. The above procedure was performed at the Korea Plant Extract Bank, and a sample of 20 mg/tube was dissolved in DMSO to a concentration of 50 mg/ml for use in cell experiments, and this was diluted with a cell culture medium and used.

Example 2: Antibacterial effect of plant extracts of the genus fluff

In order to evaluate the oral antibacterial activity of the extracts of fluff, bovine ragweed, and plant extracts of the genus fluff, P. gingivalis (Prophyromonas gingivalis; P. gingivalis, ATCC33277) was used. Porphyromonas gingivalis was activated using BHI broth containing hemin and menadione as a medium, and after culturing for 72 hours in a 37 ° C, CO₂ incubator, each well of a 96-well plate ( well) of 90 μL of bacteria, and 10 μL of the fluff extract prepared in Example 1 was added. 100 μL of the control bacteria was used, and 100 μL of BHI broth was additionally added to each well so that the final concentrations of the fluff plant extracts were 125 μg/mL, 250 μg/mL and 500 μg/mL. Thereafter, the samples were incubated for 72 hours. Absorbance was measured at 600 nm with an ELISA kit (Spectra MAX250, Molecular Devices Co.) to determine the concentration at which the growth of bacteria was inhibited.

As a result, as can be seen in FIG. 1 , it was confirmed that the growth of Porphyromonas gingivalis was inhibited when the fluff, bovine ginseng sprouts, and chrysanthemum extracts were treated. Specifically, in the case of fluff extract, it was confirmed that when 125 μg/mL was administered, it showed a higher antibacterial effect than when 250 μg/mL was administered, and in the case of bovine champignon extract and linseed grass extract, the concentration-dependent higher antibacterial effect It was confirmed that the

Example 3: Osteoclast inhibitory effect of fluff extract

Purchase a 5-week-old male ICR mouse, take the fermur and tibia, wash them with α-MEM containing antibiotics (100 units/mL penicillin and 100 μg/mL streptomycin) to obtain bone marrow cells ) was obtained. The bone marrow cells were cultured in α-MEM containing 10% fetal bovine serum (FBS) and macrophage colony-stimulating factor (M-CSF; 10 ng/mL) for 1 day. Nonadherent bone marrow cells were dispensed in a Petri dish and cultured for 3 days in the presence of M-CSF (30 ng/mL). After washing the non-adherent cells, the adherent cells were used as bone marrow-derived macrophages (BMM). In order to differentiate BMM cells into osteoclasts, BMM cells (1 × 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 Example The fluff extract prepared in 1 was treated with different concentrations (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 only RANKL were used. After culturing 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 ensure permeability. The osteoclasts were stained red by treatment with TRAP solution (Sigma Aldrich, USA). Then, to measure the activity of tartrate-resistant acid phosphatase (TRAP), a differentiation marker of osteoclasts, 100 μL of TRAP buffer (100 mM sodium citrate pH 5.0, 50 mM sodium tartrate, 3 mM p-nitrophenyl phosphate) was added and 37 After reacting at ℃ for 5 minutes, 100 μl of 0.1 N NaOH was added to terminate the reaction. Then, by measuring the absorbance at 405 nm, TRAP activity was measured.

As a result, as can be seen in Figure 2, when the fluff extract was treated on the osteoclasts induced by RANKL, TRAP activity was inhibited in a concentration-dependent manner, and, as can be seen in Figure 3, the number of osteoclasts stained red was significantly increased. It was confirmed to decrease significantly.

Through this, it was confirmed that the fluff extract exhibits an excellent osteoclast differentiation inhibitory effect and can be effective in the treatment of oral diseases and bone diseases.

Example 4: Osteoblast promoting effect of fluff extract

In order to confirm the osteoblast promoting effect of the fluff extract, after administration of the fluff extract, ALP activity and osteoblast staining were performed.

More specifically, the degree of promotion of osteoblast differentiation was observed by measuring the ALP activity of the expression level of Alkaline Phosphatase (ALP), an early biomarker of osteoblast differentiation. The myoblast cell line, C2C12 cells, purchased from the American Type Culture Collection (VA, USA) was grown with a growth medium (GM; Dulbecco's Modified containing 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin). Eagle's Medium (DMEM)). Cells were seeded, and after 1 day, the medium was replaced with a differentiation medium (DM; DMEM containing 5% FBS and 50 ng/ml rhBMP-2) to differentiate the cells. At this time, the osteoblast differentiation promoting efficacy of the extract was evaluated using a differentiation medium with or without the extract. After 3 days, the cells were washed twice with phosphate-buffered saline (PBS), and 100 μl of 1-Step PNPP (p-nitrophenyl phosphate) substrate solution (Thermo Fisher Scientific, Mass., USA) was added. Thereafter, the cells were incubated for 15 minutes at room temperature, and 50 μl of 2N NaOH was added to stop the reaction. Then, the absorbance was measured at 405 nm using a microplate reader (Wallac EnVision, PerkinElmer, MA, USA).

In addition, the osteoblast differentiation promoting efficacy of the extract was evaluated through ALP staining. Cells were prepared in the same manner as above, and after treatment with the extract, 3 days later, the cells were washed twice with PBS, fixed with 10% formalin in PBS for 30 seconds, rinsed with deionized water, Staining was performed using an Alkaline Phosphatase Kit (Sigma, MO, USA). The degree of promotion of osteoblast differentiation can be confirmed through the degree of strong staining of ALP.

As a result, as can be seen in FIG. 4 , when the fluff extract was treated on osteoblasts, ALP activity was promoted in a concentration-dependent manner, and, as shown in FIG. 5 , it was confirmed that the number of stained osteoblasts was significantly increased. .

Through this, it was confirmed that the fluff extract exhibits an excellent osteoblast differentiation promoting effect and can be effective in the treatment of oral diseases and bone diseases.

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

Claims (12)

As a pharmaceutical composition for the prevention or treatment of oral disease or bone disease comprising a fluff extract as an active ingredient,
Prevention or treatment of the oral disease is achieved through inhibition of Porphyromonas gingivalis activity,
The oral disease is periodontitis or gingivitis,
The bone disease is osteoporosis, osteomalacia, osteopenia, bone atrophy, periodontal disease, osteolysis, fibrousdysplasia, Paget's disease. disease), bone hypoplasia (osteogenesis imperfect), hypercalcemia (hypercalcemia), any one or more selected from the group consisting of, the pharmaceutical composition.
delete The pharmaceutical composition according to 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.
delete delete The pharmaceutical composition of claim 1, wherein the bone disease is osteoporosis.
delete The pharmaceutical composition of claim 1, wherein the prevention or treatment of bone disease is achieved through inhibition of osteoclast differentiation or promotion of osteoblast differentiation.
As a food composition for the prevention or improvement of oral disease or bone disease comprising a fluff extract as an active ingredient,
Prevention or treatment of the oral disease is achieved through inhibition of Porphyromonas gingivalis activity,
The oral disease is periodontitis or gingivitis,
The bone disease is osteoporosis, osteomalacia, osteopenia, bone atrophy, periodontal disease, osteolysis, fibrousdysplasia, Paget's disease. disease), bone hypoplasia (osteogenesis imperfect), hypercalcemia (hypercalcemia), any one or more selected from the group consisting of, the food composition.
As a quasi-drug composition for the prevention or improvement of oral disease or bone disease comprising fluff extract as an active ingredient,
Prevention or treatment of the oral disease is achieved through inhibition of Porphyromonas gingivalis activity,
The oral disease is periodontitis or gingivitis,
The bone disease is osteoporosis, osteomalacia, osteopenia, bone atrophy, periodontal disease, osteolysis, fibrousdysplasia, Paget's disease. disease), bone hypoplasia (osteogenesis imperfect), hypercalcemia (hypercalcemia), any one or more selected from the group consisting of, quasi-drug composition.
As a dental composition for preventing or improving oral disease or bone disease comprising a fluff extract as an active ingredient,
Prevention or treatment of the oral disease is achieved through inhibition of Porphyromonas gingivalis activity,
The oral disease is periodontitis or gingivitis,
The bone disease is osteoporosis, osteomalacia, osteopenia, bone atrophy, periodontal disease, osteolysis, fibrousdysplasia, Paget's disease. disease), bone hypoplasia (osteogenesis imperfect), hypercalcemia (hypercalcemia) will be any one or more selected from the group consisting of, a dental composition.
An antibacterial composition for Porphyromonas gingivalis comprising a fluff extract as an active ingredient.

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