Compositions containing horse chestnut extract

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating periodontal disease, which contains a horse chestnut extract as an active ingredient, and a food composition for preventing or ameliorating periodontal disease, which contains a horse chestnut extract as an active ingredient, wherein the horse chestnut extract contains avicularin and juglanin as active ingredients.

Description

[Technical field]

[0001] The present invention relates to a pharmaceutical composition for preventing or treating periodontal disease, which contains an extract of horse chestnut as an active ingredient, and a food composition for preventing or ameliorating periodontal disease, which contains an extract of horse chestnut as an active ingredient. [Background technology]

[0002] Periodontitis is a type of inflammation that occurs in the supporting tissues around the teeth due to toxins, which are metabolic products of microorganisms living in the oral cavity. Periodontitis starts with gingivitis in the early stages, and if left untreated, it progresses to periodontitis, which is accompanied by swelling, bleeding, and severe bad breath. If periodontitis continues to progress, the collagen that supports the periodontal ligament is destroyed, the alveolar bone that supports the teeth is dissolved, the periodontal ligament is peeled off, periodontal pockets are formed, and in severe cases, it leads to progressive periodontitis, which damages the teeth.

[0003] The causes of periodontal disease vary with gender, race, and age. Periodontal disease may worsen and remit over the years, and infection rates are high in patients with systemic diseases such as diabetes, AIDS, neutropenia, and Down's syndrome.

[0004] The most effective way to prevent periodontal disease is by using substances that inhibit the activity of these microorganisms and promote blood flow to the inflamed gums. However, among the drugs used to prevent and / or treat periodontal disease, antibiotics such as penicillin, erythromycin, and tetracycline are effective in killing bacteria and inhibiting their growth, but have significant side effects such as the appearance of resistant bacteria, and antibacterial agents such as chlorhexidine are effective in inhibiting the formation of dental caries, but have the problem of being highly toxic. Therefore, it is necessary to research natural products with fewer side effects.

[0005] Furthermore, with the widespread use of dental implants, various diseases related to the implants are occurring frequently.

[0006] Of these diseases, peri-implantitis is a disease that appears around the implant, not in the natural teeth, and is therefore the disease that requires the most attention after implantation. It has been reported that peri-implantitis occurs in more than 60% of implants that have been performed so far. In particular, unlike natural teeth, implants do not have nerves distributed inside them, so there is a problem that the rapid increase of peri-implantitis is difficult to notice in the early stages of its occurrence, and is only recognized after the symptoms have seriously worsened.

[0007] However, if left untreated in the early stages of its development, peri-implantitis can even lead to alveolar bone destruction in severe cases. Therefore, careful management of peri-implantitis is required, and severe bone loss around the implant may even lead to reoperation due to implant failure.

[0008] Peri-implantitis also tends to progress more quickly and be more severe than other diseases that occur in natural teeth.

[0009] However, existing drugs used for preventing or treating peri-implantitis do not effectively or fundamentally treat peri-implantitis, but merely alleviate its symptoms. Therefore, when peri-implantitis occurs, it is difficult to treat peri-implantitis with drugs alone, and instead, other methods such as surgical treatment, operation, etc. have been mainly used.

[0010] Therefore, there is a need to develop drugs that can effectively prevent or treat peri-implantitis. [Prior art documents] [Patent documents]

[0011] [Patent Document 1] Korean Patent Registration No. 10-2260037 Summary of the Invention [Problem to be solved by the invention]

[0012] The present invention provides a pharmaceutical composition for preventing or treating periodontal disease and a food composition for preventing or ameliorating periodontal disease, which are safe because they have no side effects even when administered over a long period of time and have low cytotoxicity, and which suppress periodontal disease by inhibiting osteoclast differentiation. [Means for solving the problem]

[0013] The present inventors have recognized the need for research into the prevention or treatment of periodontal disease and have set out to achieve this objective of researching natural substances that can prevent or treat such diseases.

[0014] Specifically, the present inventors have completed the present invention while researching natural products that have no toxicity or side effects even when administered for a long period of time, effectively inhibit the differentiation of osteoclasts to reduce bone loss, and can prevent / treat the onset of periodontal disease induced by or caused by bone loss. It has been confirmed that an extract from Aesculus turbinata has a preventive and / or therapeutic effect against periodontal disease by inhibiting tartrate-resistant acid phosphatase (hereinafter referred to as "TRAP").

[0015] The present invention will be described in detail below.

[0016] Pharmaceutical composition for preventing or treating periodontal disease containing horse chestnut extract as an active ingredient The present invention provides a pharmaceutical composition comprising a horse chestnut extract as an active ingredient for preventing or treating periodontal disease, and comprising avicularin and juglanin as active ingredients of the horse chestnut extract for preventing or treating periodontal disease.

[0017] In the present invention, the term "Horse Chestnut" refers to a plant belonging to the family Hippocastanaceae, and also known as Conker Tree and Marronier in France.

[0018] In an exemplary embodiment of the present invention, the horse chestnut extract may be obtained from the leaves or seeds (Saraja), especially from the leaves.

[0019] For example, the horse chestnut extract of the present invention may be used by drying commercially available horse chestnut leaves or leaves harvested from spring to autumn in the shade. Alternatively, the horse chestnut extract may be a dried horse chestnut extract prepared from horse chestnut seeds, and may contain 10% by weight or more of triterpene glycoside, i.e., anhydrous aescin. More preferably, an extract of horse chestnut leaves may be used.

[0020] In an exemplary embodiment of the present invention, the horse chestnut may be, but is not limited to, Aesculus turbinata BLUME, Aesculus chinensis Bge and Aesculus wilculus Rehd, Aesculus hippocastanum L., and the like, in particular Aesculus hippocastanum L. Kalisz, Poland.

[0021] In the present invention, the term "extract" refers to a crude extract as commonly used in the art, but in a broader sense also includes fractions obtained by further fractionating the extract, and specifically, the present invention includes an extract obtained by extraction, a dilution or concentrate of the extract, a dried product obtained by drying the extract, a crude product, a purified product, or a mixed product of the extract, an extract of any preparation that can be formed using the extract, and the extract itself.

[0022] In the present invention, the term "active ingredient" refers to a substance or group of substances (including herbal medicines whose pharmacologically active ingredients have not been identified) that are expected to directly or indirectly exert the efficacy of the composition due to their inherent pharmacological action, and is meant to include the main ingredient.

[0023] In the present invention, the term "periodontal disease" refers to a disease that appears in the tissues surrounding the teeth, such as the gums, periodontal ligament, and alveolar bone.

[0024] In the present invention, the term "prevention" refers to any action of suppressing or delaying periodontal disease by inhibiting TRAP, an osteoclast differentiation marker, through administration of the Horse Chestnut extract of the present invention.

[0025] In the present invention, the term "treatment" refers to any action of improving or beneficially altering periodontal disease by inhibiting TRAP, an osteoclast differentiation marker, through administration of the Horse Chestnut extract of the present invention.

[0026] In the present invention, the horse chestnut extract contains avicularin and jugranin as active ingredients.

[0027] Avicularin, the active ingredient of the horse chestnut extract of the present invention, can be represented by the following chemical formula 1.

[0028] [ka]

[0029] Jugranin, which is the active ingredient of the horse chestnut extract of the present invention, can be represented by the following chemical formula 2.

[0030] [ka]

[0031] In the present invention, the Horse Chestnut extract may contain avicularin as an active ingredient in an amount of 0.1 to 4.0% by weight, particularly 0.3 to 3.0% by weight, and further particularly 0.5 to 2.0% by weight, based on the total weight of the Horse Chestnut extract.

[0032] In the present invention, the Horse Chestnut extract may contain jugranin as an active ingredient in an amount of 0.1 to 3.0% by weight, particularly 0.12 to 2.0% by weight, and further particularly 0.15 to 1.0% by weight, based on the total weight of the Horse Chestnut extract.

[0033] In the present invention, the horse chestnut extract may further contain quercitrin and afzelin as further active ingredients.

[0034] Quercitrin, the active ingredient of the horse chestnut extract of the present invention, can be represented by the following chemical formula 3.

[0035] [ka]

[0036] Afzelin, the active ingredient of the horse chestnut extract of the present invention, can be represented by the following chemical formula 4.

[0037] [ka]

[0038] In the present invention, the horse chestnut extract may contain quercitrin as an active ingredient in an amount of 1.0 to 7.0% by weight, particularly 1.3 to 6.0% by weight, and further particularly 1.5 to 5.0% by weight, based on the total weight of the horse chestnut extract.

[0039] In the present invention, the Horse Chestnut extract may contain afzelin as an active ingredient in an amount of 0.1 to 3.0% by weight, particularly 0.2 to 2.0% by weight, and further particularly 0.3 to 1.0% by weight, based on the total weight of the Horse Chestnut extract.

[0040] That is, the horse chestnut extract may contain, as active ingredients based on the total weight of the horse chestnut extract, 0.1 to 4.0% by weight of avicularin, 1.0 to 7.0% by weight of quercitrin, 0.1 to 3.0% by weight of jugranin, and 0.1 to 3.0% by weight of afzelin, and in particular may contain (1) 0.3 to 3.0% by weight of avicularin, 1.3 to 6.0% by weight of quercitrin, 0.12 to 2.0% by weight of jugranin, and 0.2 to 2.0% by weight of afzelin, or (2) 0.5 to 2.0% by weight of avicularin, 1.5 to 5.0% by weight of quercitrin, 0.15 to 1.0% by weight of jugranin, and 0.3 to 1.0% by weight of afzelin.

[0041] In the following exemplary embodiments, unless otherwise specified, the horse chestnut extract of the present invention may contain avicularin, quercitrin, jugranin and afzelin in the above weight percentages.

[0042] In the present invention, the contents of avicularin, quercitrin, jugranin and afzelin in a horse chestnut extract can be measured by quantifying the contents of avicularin, quercitrin, jugranin and afzelin contained in the extract.

[0043] Since avicularin, quercitrin, jugranin, and afzelin contained in the horse chestnut extract of the present invention may be converted into other substances during the quantification process, the quantification process must be strictly controlled in order to accurately quantify the components in the horse chestnut extract.

[0044] In particular, when the horse chestnut extract of the present invention contains avicularin and jugranin in a weight ratio within the above range, the effect of preventing or treating periodontal disease will be excellent.

[0045] Specifically, avicularin and jugranin as active ingredients of the horse chestnut extract are novel ingredients that have never been disclosed as active ingredients of the horse chestnut extract, and avicularin and jugranin can be considered as functional ingredients that inhibit the differentiation of osteoclasts.

[0046] However, the contents of avicularin and jugranin in the horse chestnut extract may vary depending on the region, year, and harvest time, within the range in which the effect is maintained.

[0047] The pharmaceutical composition of the present invention is effective in preventing and treating periodontal disease by inhibiting TRAP, an osteoclast differentiation marker, and has excellent pharmacological effects with few side effects.

[0048] Specifically, in bone metabolism and bone formation, bone homeostasis is maintained by a balance between bone formation by osteoblasts and bone resorption by osteoclasts. Osteoclasts attach to the bone surface and secrete acids and proteolytic enzymes to remove the bone matrix that constitutes the bone and dissolve aged bone.

[0049] When osteoclast differentiation is activated, various enzymes that degrade bone are produced. TRAP, an acid phosphatase enzyme that oxidizes and degrades bone, is used as a specific marker for osteoclasts.

[0050] Specifically, the secretion of TRAP increases during bone resorption by osteoclasts, and its activity increases in the presence of ATP and nitrophenyl phosphate. TRAP is a cytochemical marker enzyme that can measure the degree of osteoclast differentiation.

[0051] Therefore, a method for evaluating the inhibitory effect on osteoclast differentiation is to analyze TRAP activity, and it can be confirmed that the pharmaceutical composition of the present invention can prevent bone destruction by inhibiting TRAP activity.

[0052] In another embodiment, the active ingredient may be 0.5 to 2.0 g of avicularin based on 100 g of the horse chestnut extract.

[0053] In another embodiment, the active ingredient may contain 1.5 to 5.0 g of quercitrin based on 100 g of the horse chestnut extract.

[0054] In another embodiment, 0.15 to 1.0 g of jugranin may be contained as an active ingredient based on 100 g of the horse chestnut extract.

[0055] In another embodiment, the active ingredient may contain 0.3 to 1.0 g of afzelin based on 100 g of the horse chestnut extract.

[0056] In the present invention, the horse chestnut extract can be obtained by an extraction process including extracting horse chestnut leaves with a 10% to 50% (v / v) aqueous alcohol solution and concentrating the extract.

[0057] In the present invention, the term "alcohol" refers to a compound having a hydroxyl group attached to a carbon atom of an alkyl or substituted alkyl, the term "alkyl" refers to a straight chain saturated hydrocarbon group or a branched saturated hydrocarbon group, and the term "substituted alkyl group" means that the substituent attached to a carbon of the alkyl group is hydroxy, cyano, halide, etc.

[0058] In an exemplary embodiment of the present invention, the aqueous alcohol may be a 20-30% (v / v) aqueous alcohol solution.

[0059] The aqueous alcohol solution refers to a C1 to C4 alcohol including a 20 to 30% (v / v) aqueous ethanol solution, an aqueous methanol solution, etc., preferably an aqueous ethanol solution or an aqueous methanol solution, and more preferably a 25% (v / v) aqueous ethanol solution.

[0060] In the present invention, the term "C1-C4" means a functional group or main chain having 1 to 4 carbon atoms.

[0061] In the present invention, the extract can be dried using hot air drying, freeze drying, or spray drying. For example, the extract can be dried with hot air using a hot air dryer, or the extract can be frozen and freeze-dried by lowering the air pressure, or the extract can be sprayed and then spray-dried by blowing hot air onto it.

[0062] In preparing the Horse Chestnut extract of the present invention, dried Horse Chestnut leaves, non-dried Horse Chestnut leaves, or mixtures thereof can be used. For effective extraction, Horse Chestnut leaves can be used in a finely ground form.

[0063] In the present invention, the alcohol extract can be obtained by the following steps, but is not limited thereto. (S1) an extraction step in which horse chestnut leaves are extracted with a 10% to 50% (v / v) aqueous alcohol solution at 75 to 85°C for 1 to 5 hours to obtain an alcohol extract; (S2) concentrating the alcohol extract under a temperature condition of 25 to 35° C. and a pressure condition of −755 to −650 mmHg to obtain a concentrated extract; and (S3) A process of adding purified water to the concentrated extract so that the Brix concentration becomes 20 to 40%, dissolving the concentrated extract, and then freeze-drying the dissolved concentrated extract to obtain a horse chestnut extract.

[0064] In the present invention, the alcohol extract of (S1) can be prepared using a 10 to 50% (v / v) alcohol aqueous solution, preferably a 20 to 30% (v / v) alcohol aqueous solution, which is 5 to 15 times (v / w) the volume of the horse chestnut leaves, and step (S1) can be repeated 1, 2 or 3 times. For example, when repeated 2 or 3 times, the alcohol extract of each step is obtained, separated, and finally mixed to obtain the alcohol extract.

[0065] The pharmaceutical composition for preventing or treating periodontal disease may contain, as an active ingredient, a Horse Chestnut extract in an amount of 10 to 90% by weight based on the total weight, particularly 15 to 70% by weight of the Horse Chestnut extract, further 40 to 60% by weight of the Horse Chestnut extract, and even more particularly 45 to 55% by weight of the Horse Chestnut extract.

[0066] In the present invention, the periodontal disease may be one or more selected from the group consisting of gingivitis, periodontitis, alveolar pyorrhea, and peri-implantitis.

[0067] In the present invention, the term "gingivitis" refers to the gingiva, i.e. inflammation of the gums, which can be classified into simple gingivitis, ulcerative gingivitis, gangrenous gingivitis, and hypertrophic gingivitis.

[0068] In the present invention, the term "periodontitis", also called paradentitis, refers to an inflammatory disease affecting the periodontal tissues.

[0069] In the present invention, the term "pyorrhea alveolar" refers to chronic marginal periodontitis, which is a symptom of purulent periodontitis.

[0070] In the present invention, the term "peri-implantitis" refers to an inflammatory disease occurring in the gingival and bone tissue surrounding a dental implant.

[0071] The pharmaceutical composition for preventing or treating periodontal disease containing the Horse Chestnut extract of the present invention contains the Horse Chestnut extract as an active ingredient, and may further contain a pharma- ceutically acceptable carrier for formulating by conventional methods into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and the like, as well as for topical applications and sterile injectable preparations.

[0072] In the present invention, the term "pharmaceutical acceptable" refers to a composition that is physiologically tolerable and that does not normally cause allergic reactions, such as gastrointestinal upset, dizziness, or similar reactions, when administered to a human.

[0073] In the present invention, the term "pharmaceutical acceptable carrier" typically includes liquid or non-liquid bases of pharmaceutical compositions. When pharmaceutical compositions are provided in liquid form, the carrier typically includes pyrogen-free water; isotonic saline or buffer (water) solutions, such as phosphate, citrate, etc. The injection buffer may be hypertonic, isotonic, or hypotonic in a particular reference medium, i.e., the buffer may have a high, equal, or low salt content in a particular reference medium, and preferably, the aforementioned salts may be used in concentrations that do not induce cell damage due to osmotic or other concentration effects.

[0074] The reference medium may be present in the "in vivo" method, such as blood, lymph, cytoplasmic fluid, or other body fluid, or a body fluid that may be used as a reference medium (e.g., a common buffer or liquid) in the "ex vivo" method. Such common buffers or liquids are known to those skilled in the art.

[0075] Pharmaceutically acceptable carriers may include those commonly used in the art, such as, but not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0076] Furthermore, the pharmaceutical composition of the present invention may contain diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants, and other pharma- ceutically acceptable additives.

[0077] The pharmaceutical composition of the present invention can be prepared in the dosage form of a solution, suspension, powder, granules, tablets, capsules, pills or extract.

[0078] The compositions of the present invention can be administered orally or parenterally (eg, as a liniment or by intravenous, subcutaneous or intraperitoneal injection).

[0079] In the present invention, the term "oral administration" refers to a method of taking a drug through the mouth to relieve a pathological symptom (periodontal disease), and in the present invention, the term "parenteral administration" refers to a method of subcutaneous, intramuscular, intravenous, or intraperitoneal administration using a tube, excluding administration through the mouth.

[0080] Solid dosage forms for oral administration may include powders, granules, tablets, capsules, soft capsules, pills, etc. Liquid dosage forms for oral administration may include suspensions, oral liquids, emulsions, syrups, aerosols, etc., and may contain various additives such as moisturizers, sweeteners, flavorings, preservatives, etc., in addition to water and liquid paraffin, which are often used as simple diluents.

[0081] As the dosage form for parenteral administration, external preparations such as sterile aqueous solutions, liquids, non-aqueous solvents, suspensions, emulsions, eye drops, eye ointments, syrups, suppositories, and aerosols, as well as sterile injections, can be formulated by conventional methods and used, preferably creams, gels, patches, sprays, ointments, plasters, lotions, liniments, eye ointments, eye drops, pastes, or poultices, but are not limited thereto. Compositions for topical administration may be anhydrous or aqueous, depending on the clinical prescription. As non-aqueous solvents and suspending agents, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate can be used. As the base for suppositories, witepsol, macrogol, Tween 61, cocoa butter, laurin, glycerogelatin, and the like can be used.

[0082] The pharma- ceutically acceptable additive of the present invention may be included in an amount of 0.1 to 99.9 parts by weight, specifically 0.1 to 50 parts by weight, based on the composition, but is not limited to this range.

[0083] In the present invention, the pharmaceutical composition for preventing or treating periodontal disease, which contains an Aesculus turbinata extract as an active ingredient, exhibits an inhibitory effect against TRAP, and thus has an effect of preventing or treating periodontal disease. The pharmaceutical composition for preventing or treating periodontal disease, which contains an Aesculus turbinata extract as an active ingredient, can be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, intraocularly or intradermally, as in the conventional manner, and can be administered in particular by the oral route.

[0084] The pharmaceutical composition of the present invention for preventing or treating periodontal disease containing a horse chestnut extract as an active ingredient is administered once or several times a day, preferably at a dose of 0.001 mg / kg to 50 mg / kg.

[0085] Food composition containing horse chestnut extract as an active ingredient Provided is a food composition containing a horse chestnut extract as an active ingredient, the composition containing avicularin and jugranin as active ingredients for preventing or improving periodontal disease.

[0086] In the present invention, the horse chestnut extract contains, as active ingredients, 0.6 to 1.5% by weight of avicularin and 0.2 to 0.4% by weight of jugranin based on the total weight of the horse chestnut extract.

[0087] The horse chestnut extract and the method for preparing it are the same as described above.

[0088] The food composition is for improving or preventing periodontal disease.

[0089] In the present invention, the term "amelioration" refers to any effect that at least reduces the severity of the symptoms of periodontal disease.

[0090] In the food composition of the present invention, when the horse chestnut extract is used as an additive in the food composition, it can be added as it is or can be used in combination with other foods or food ingredients, and can be used appropriately in a conventional manner. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use, such as prevention, health, or treatment.

[0091] The food composition of the present invention can be formulated in any of the forms of common foods and beverages, such as powders, granules, pills, tablets, and capsules.

[0092] The type of food is not particularly limited, and examples of foods to which the substance can be added include meats, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream, various soups, beverages, tea, alcoholic beverages, and multivitamin supplements, and can include any food in the conventional sense.

[0093] In general, in the manufacture of food and drink, the horse chestnut extract may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on 100 parts by weight of raw material. However, when taking it for a long time for health hygiene or health management, the amount may be less than the above range. Furthermore, since the present invention uses an extract from a natural product and there is no problem in terms of safety, it can be used in an amount exceeding the above range.

[0094] The beverage in the food composition of the present invention may contain various flavorings or natural carbohydrates as additional components, similar to conventional beverages. Examples of natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame can be used. The proportion of natural carbohydrates may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the beverage of the present invention.

[0095] In addition to the above, the food composition of the present invention contains various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonation agents used in carbonated drinks. Furthermore, the food composition of the present invention may contain natural fruit juice and fruit pulp to produce fruit juice drinks and vegetable drinks. These ingredients may be used alone or in combination. The proportion of these additives is not particularly limited, but is usually selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the food composition of the present invention.

[0096] Methods of prevention or treatment Another object of the present invention is to provide a method for preventing or treating periodontal disease by administering to a subject in need of treatment a therapeutically effective amount of a pharmaceutical composition comprising an extract of Aesculus turbinata as an active ingredient.

[0097] In an embodiment of the present invention, the term "subject in need of treatment" refers to a mammal, including a human, and the term "administration" refers to providing a given substance to a patient by any suitable method. In the treatment method of the present invention, a pharmaceutical composition for preventing or treating periodontal disease, which contains an extract of Aesculus turbinata as an active ingredient, can be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, intraocularly or intradermally in a conventional manner.

[0098] In the method for preventing or treating periodontal disease according to the invention, the horse chestnut extract of the invention can be administered conventionally by oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes, and in particular can be administered orally.

[0099] In an embodiment of the present invention, the term "therapeutically effective amount" refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal, or human, as considered by a researcher, veterinarian, physician, or other clinician, including an amount that induces an improvement in the symptoms of the disease or disorder being treated. It is clear to those skilled in the art that the therapeutically effective dosage and frequency of administration of the active ingredient of the present invention will vary depending on the desired effect. Therefore, the optimal dosage can be easily determined by those skilled in the art and can be adjusted according to various factors, including the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, the type of formulation, and the age, weight, health condition, sex, and diet of the patient, the administration time, administration route, and excretion rate of the composition, the treatment period, and concomitant drugs.

[0100] In the method for preventing or treating periodontal disease of the present invention, the horse chestnut extract of the present invention is administered once or several times a day, preferably at a dose of 0.001 mg / kg to 50 mg / kg for adults.

[0101] Use of a pharmaceutical composition containing a horse chestnut extract as an active ingredient for the prevention or treatment of periodontal disease It is an object of the present invention to provide the use of a pharmaceutical composition comprising an extract of Aesculus turbinata as an active ingredient for preventing or treating periodontal disease.

[0102] Use of a pharmaceutical composition comprising an extract of Aesculus turbinata as an active ingredient for the manufacture of a medicament for the prevention or treatment of periodontal disease It is an object of the present invention to provide the use of a pharmaceutical composition comprising an extract of Aesculus turbinata as an active ingredient for the manufacture of a medicament for the prevention or treatment of periodontal disease.

[0103] In an exemplary embodiment of the present invention, a pharmaceutical composition containing the horse chestnut extract as an active ingredient for the manufacture of a medicament can be mixed with an acceptable carrier, etc., and can further contain other medicinal agents.

[0104] Other daily necessities containing a pharmaceutical composition for preventing or treating periodontal disease, which contains an extract of horse chestnut as an active ingredient. It is an object of the present invention to provide a toothpaste containing a pharmaceutical composition comprising a horse chestnut extract as an active ingredient for the prevention or treatment of periodontal disease.

[0105] In the present invention, when the pharmaceutical composition for preventing or treating periodontal disease is used as a toothpaste, the composition may be added to the toothpaste as it is, or may be mixed with or added to a functional food or functional ingredient, and may be used appropriately in a conventional manner. The amount of the active ingredient to be added may be appropriately determined depending on the purpose of use.

[0106] Specifically, the additives may include additives that are commonly used in toothpastes, including fluorine compounds, humectants, foaming agents, binders, sweeteners, abrasives, flavoring agents, preservatives, etc., and the specific types of additives are not limited as long as they can be generally used in various ways in toothpastes.

[0107] Another object of the present invention is to provide a mouthwash comprising a pharmaceutical composition for preventing or treating periodontal disease.

[0108] The mouthwash includes a composition for rinsing the oral cavity, a fluorine compound for cleaning the oral cavity and for antibacterial action, a humectant, a foaming agent, a binder, a sweetener, an abrasive, a flavoring agent, a preservative, etc., and the specific type is not limited as long as it is one that can be used in a general mouthwash. A specific formulation is defined as including both a composition and a mixing range that can be selected and mixed by a person skilled in the art.

[0109] Another object of the present invention is to provide an oral spray comprising a pharmaceutical composition for the prevention or treatment of periodontal disease.

[0110] The mouth spray may contain all compositions and other agents for cleaning and antibacterial action in the mouth. A specific formulation is defined to include all compositions and mixing ranges that can be selected and mixed by one skilled in the art.

[0111] The pharmaceutical compositions, methods of treatment, uses, and other daily products of the present invention are equally applicable unless they contradict each other.

[0112] The advantages and features of the present invention, as well as the methods for achieving them, will become apparent with reference to the exemplary embodiments described in detail below. However, the present invention is not limited to the exemplary embodiments disclosed below, but may be embodied in various different forms. These exemplary embodiments are provided merely to enhance the disclosure of the present invention and to fully convey the scope of the present invention to those skilled in the art to which the present invention pertains, and the present invention is defined solely by the claims. Effect of the Invention

[0113] The horse chestnut extract of the present invention is safe with low cytotoxicity and exhibits an inhibitory effect on osteoclast differentiation.

[0114] Therefore, the composition of the present invention containing a horse chestnut extract as an active ingredient may be useful for preventing or treating periodontal disease, has significantly fewer side effects, and has the effect of preventing or treating periodontal disease. [Brief description of the drawings]

[0115] [Figure 1] FIG. 1 shows the results of high performance liquid chromatography (hereinafter referred to as "HPLC") of the horse chestnut extract prepared according to Example 1 of the present invention. [Diagram 2] FIG. 2 shows the HPLC results of avicularin. [Diagram 3] FIG. 3 shows the HPLC results of quercitrin. [Figure 4] FIG. 4 shows the HPLC results of jugranin. [Diagram 5] FIG. 5 shows the HPLC results of afzelin. [Figure 6] FIG. 6 shows the results of a cytotoxicity assay of the horse chestnut extract prepared according to Example 1 of the present invention. [Figure 7]FIG. 7 shows the results of a cytotoxicity assay of avicularin. [Figure 8] FIG. 8 shows the results of a cytotoxicity assay of quercitrin. [Figure 9] FIG. 9 shows the results of a cytotoxicity assay of jugranin. [Figure 10] FIG. 10 shows the results of a cytotoxicity assay of afzelin. [Figure 11] FIG. 11 shows the inhibition of TRAP activity by the Horse Chestnut extract prepared according to Example 1 of the present invention. [Figure 12] FIG. 12 shows the inhibition of TRAP activity by avicularin. [Figure 13] FIG. 13 shows the inhibition of TRAP activity by quercitrin. [Figure 14] FIG. 14 shows the inhibition of TRAP activity by juglanin. [Figure 15] FIG. 15 shows the inhibition of TRAP activity by afzelin. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0116] Best Mode for Carrying Out the Invention The terms used in this specification are selected as widely used as possible while taking into consideration the functions of the present invention, but may change depending on the intentions of those skilled in the art, precedents, the emergence of new technologies, etc. Furthermore, in certain cases, there are terms arbitrarily selected by the applicant, and the meanings of such terms will be explained in detail in the description of the present invention. Therefore, the terms used in this specification should be defined based on the meanings of the terms and the overall content of the present invention, rather than simply the names of the terms.

[0117] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by those skilled in the art to which the present invention pertains. Terms as defined in commonly used dictionaries shall be interpreted as having a meaning consistent with the meaning in the context of the relevant art, and shall not be interpreted in an idealized or overly formal sense unless expressly defined in this application.

[0118] Numeric ranges include the numbers defined within the range. Every maximum numerical limit given throughout this specification includes every lower numerical limit, as if a single lower numerical limit were specified. Every minimum numerical limit given throughout this specification includes every upper numerical limit, as if a single upper numerical limit were specified. Every numerical limit given throughout this specification will include every greater numerical range within the broader numerical range, as if a narrower numerical limit were specified.

[0119] In order to facilitate understanding of the present invention, examples and preparations are presented. The following examples and preparations are provided only to facilitate understanding of the present invention, and the contents of the present invention are not limited to these examples and preparations. EXAMPLES

[0120] Example 1. Preparation of Horse Chestnut Extract Horse chestnut (Aesculus hippocastanum L.) leaves from Kalisz, Poland were dried in the shade at room temperature and then crushed. 80 kg of crushed leaves were extracted with 800 L of 25% (v / v) aqueous ethanol at 80°C for 3 h and filtered through a 25 μm cartridge filter to obtain 450 L of primary extract. The residue of the horse chestnut leaves was extracted with 800 L of 25% (v / v) aqueous ethanol at 80°C for 3 hours, and filtered through a 25 μm cartridge filter to obtain 830 L of a secondary extract. The primary extract and the secondary extract (total extract: 1280 L) were mixed and then concentrated under conditions of a temperature of 75 to 85° C. and a pressure of −755 to −650 mmHg to obtain 20 L of concentrated extract. Next, 50 L of purified water was added to the concentrated extract, and the ethanol solvent was removed by vacuum concentration at 20-30°C, and 60 L of purified water was added to dissolve the extract to a Brix concentration of 25-35%. After freeze-drying using a freeze dryer, the extract was pulverized on a 0.5 mm pulverizing mesh of a pulverizer at 1,200 rpm to obtain 18 kg of freeze-dried powder of the horse chestnut extract.

[0121] Example 2. Components of Horse Chestnut Extract (1)Analysis method Whether or not the horse chestnut extract contains avicularin, quercitrin, jugranin and afzelin, particularly avicularin and jugranin which are not known to be contained as active ingredients in horse chestnut extracts, was confirmed as follows. 100 mg of the horse chestnut extract was placed in a tube, thoroughly dissolved in 100 mL of 100% methanol, filtered through a 0.22 μm filter, and subjected to HPLC. The results are shown in Figure 1. Furthermore, avicularin, quercitrin, jugranin, and afzelin were each subjected to HPLC, and the results are shown in Figures 2 to 5, respectively. The HPLC conditions were as follows. i) Column: CAPCELL PAK C18 UG120 (SHISEIDO, 4.6 x 250 mm, 5 μm) ii) Detector: UV 254 nm iii) Flow rate: 1.0mL / min iv) Mobile phase Mobile phase A: 0.1% (w / v) phosphoric acid, Mobile phase B: acetonitrile

[0122] [Table 1]

[0123] (2) Results The horse chestnut extract obtained in Example 1 was found to contain avicularin, quercitrin, jugranin, and afzelin by the above analytical method, and in particular, it was confirmed that it contained avicularin and jugranin, which are not known as active ingredients in horse chestnut extracts. These contents were as follows: Avicularin: 0.7% by weight Quercitrin: 2.2% by weight Jugranine: 0.2% by weight Afzelin: 0.4% by weight

[0124] Preparation of reagents Avicularin, quercitrin, jugranin, and afzelin were purchased from Sigma-Aldrich Korea, and other reagents were purchased from Sigma-Aldrich, Sigma-Aldrich Korea, and Duksan General Science.

[0125] Experimental Example Experimental example 1. Cytotoxicity assay (1) Experimental method Mouse macrophage cell line RAW264.7 cells were plated in a 96-well plate (Corning, USA) at a cell count of 1 × 10 3 The cells were then treated with the Aesculus turbinata extract of the present invention at concentrations of 1.25, 2.5, 5, and 10 μg / ml, respectively, and incubated under 5% CO 2 The cells were cultured in an incubator at 37°C for 4 days. Furthermore, the cells were treated with avicularin, quercitrin, jugranin, and afzelin, the main components of the horse chestnut extract, at concentrations of 2.5, 5, 10, and 25 μM, respectively, and cultured in the same manner. A total of 100 μl of medium was removed from each well, and each well was washed with 1×PBS (phosphate-buffered saline), after which 100 μl of fresh medium was added. The cells were treated with 50 μl of XTT reagent per well, and then reacted for 4 hours. The change in absorbance was measured at 490 nm and 650 nm (reference wavelength) using an ELISA (enzyme-linked immunosorbent assay) reader (Molecular Devices, USA), and the cell viability was expressed as a percentage.

[0126] (2) Results The results in Figure 6 show that the horse chestnut extract of Example 1 of the present invention showed no cytotoxicity at concentrations of 25, 50, 75, and 100 μg / mL, and further, avicularin, quercitrin, jugranin, and afzelin showed no cytotoxicity at concentrations of 2.5, 5, 10, and 25 μM, respectively (see Figures 7 to 10).

[0127] Experimental Example 2. TRAP inhibitory activity of horse chestnut extract and its main components (1) Experimental method To confirm the effect of the Aesculus turbinata extract on osteoclast differentiation, mouse macrophage cell line (hereinafter referred to as "Raw 264.7 cells") was treated with NFκB activator receptor ligand (hereinafter referred to as "RANKL") to induce osteoclast differentiation. The cells were treated with the Aesculus turbinata extract of Example 1 and the main components of the Aesculus turbinata extract, i.e., avicularin, quercitrin, jugranin, and afzelin, respectively, and the activity of TRAP, an osteoclast-specific marker, was measured. Specifically, Raw 264.7 cells were cultured at 3 × 10 in a 96-well plate. 3 The cells were plated at a density of 100 cells / well and treated with RANKL to a final concentration of 100 ng / ml, followed by treatment with the Horse Chestnut extract of Example 1 at concentrations of 1.25, 2.5, 5, and 10 μg / ml. The cells were incubated in 5% CO 2 The cells were cultured in an incubator at 37°C for 4 days. Furthermore, the plants were treated with avicularin, quercitrin, jugranin, and afzelin, which are the main components of the horse chestnut extract, at concentrations of 2.5, 5, 10, and 25 μM, respectively, and cultured in the same manner. After removing the medium, 50 μl of the extract was added to each well and reacted for 5 minutes, followed by pipetting. 50 μl of substrate solution (3,3′,5,5′-tetramethylbenzidine) was added to each well and reacted for 60 minutes at 37° C. Then, 50 μl of stop solution (0.5 N NaOH) was added to each well and absorbance was measured at 405 nm.

[0128] (2) Results As a result, as shown in Figure 11, the Horse Chestnut extract of Example 1 inhibited TRAP activity from a concentration of 1.25 μg / ml. Specifically, about 73% of the activity was inhibited at 1.25 μg / ml, 91% of the activity was inhibited at 2.5 μg / ml, and more than 98% of the activity was inhibited at 5 μg / ml. It was confirmed that the higher the concentration of the Horse Chestnut extract used, the better the TRAP inhibitory effect (see Figure 11). Furthermore, it was confirmed that avicularin, quercitrin, jugranin, and afzelin each showed inhibition of TRAP activity from 2.5 μM, but the horse chestnut extract of Example 1 showed far superior TRAP inhibitory activity compared to the single components (see Figures 12 to 15).

[0129] Therefore, the horse chestnut extract of Example 1 of the present invention can be applied as a pharmaceutical composition for preventing or treating periodontal disease and a food composition for preventing or improving periodontal disease, and can be useful for preventing, treating and / or improving periodontal disease. It is expected to prevent and / or treat periodontal disease and improve symptoms with very few side effects.

Claims

1. A pharmaceutical composition for preventing or treating periodontal disease, comprising an extract of horse chestnut as an active ingredient, the extract of horse chestnut comprising avicularin and jugranin as active ingredients.

2. 2. The pharmaceutical composition according to claim 1, wherein the Aesculus turbinata extract contains, as active ingredients, 0.1-4.0% by weight of avicularin and 0.1-3.0% by weight of jugranin, based on the total weight of the Aesculus turbinata extract.

3. 2. The pharmaceutical composition of claim 1, wherein the horse chestnut extract further comprises quercitrin and afzelin as active ingredients.

4. 4. The pharmaceutical composition according to claim 3, wherein the Aesculus turbinata extract further contains, as active ingredients, 1.0 to 7.0% by weight of quercitrin and 0.1 to 3.0% by weight of afzelin, based on the total weight of the Aesculus turbinata extract.

5. 2. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition inhibits tartrate-resistant acid phosphatase (TRAP), which is an osteoclast differentiation marker.

6. 2. The pharmaceutical composition according to claim 1, wherein the Aesculus turbinata extract is obtained by an extraction process comprising extracting Aesculus turbinata leaves with 10% to 50% (v / v) aqueous alcohol solution and concentrating.

7. 2. The pharmaceutical composition according to claim 1, wherein the content of the horse chestnut extract is 10% by weight to 90% by weight based on the total weight of the pharmaceutical composition.

8. 2. The pharmaceutical composition according to claim 1, wherein the periodontal disease is at least one selected from the group consisting of gingivitis, periodontitis, alveolar pyorrhea, and peri-implantitis.

9. A food composition comprising an extract of horse chestnut as an active ingredient, the extract of horse chestnut comprising avicularin and jugranin as active ingredients.

10. 10. The food composition according to claim 9, wherein the Aesculus turbinata extract contains, as active ingredients, 0.1 to 4.0% by weight of avicularin and 0.1 to 3.0% by weight of jugranin, based on the total weight of the Aesculus turbinata extract.

11. A toothpaste comprising the pharmaceutical composition for preventing or treating periodontal disease according to claim 1.

12. A mouthwash comprising the pharmaceutical composition for preventing or treating periodontal disease according to claim 1.

13. An oral spray comprising the pharmaceutical composition for preventing or treating periodontal disease according to claim 1.

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