KR20220089042A - Composition of preventing or improving bone disease comprising Vitex rotundifolia extract - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating bone disease, and a food composition for improving bone disease, comprising an extract of Vitex rotundifolia as an active ingredient. As it exhibits an excellent effect on inhibition, it can be used as a pharmaceutical composition or a food composition useful for improving, preventing or treating various bone diseases.

Description

A composition for preventing or improving bone disease, comprising an extract of Sunbigi tree {Composition of preventing or improving bone disease comprising Vitex rotundifolia extract}

The present invention relates to a pharmaceutical composition for preventing or treating bone disease, comprising an extract of Vitex rotundifolia as an active ingredient, and a food composition for improving bone disease.

In the metabolism of bone, bone is destroyed, resorbed, or new bone is formed by the activity of osteoblasts and osteoclasts. In a normal person, bone remodeling occurs by the interaction between osteoblasts and osteoclasts. However, the anti-acid properties of bone resorption and bone formation are more likely to be out of balance due to rapid bone loss after menopause in women.

Osteoporosis initially causes bone loss due to bone resorption, which is determined by the ability of osteoblasts to differentiate into osteoblasts because bone formation is insufficient. Therefore, it is necessary to reduce bone resorption by promoting the differentiation of osteoblasts or inhibiting the differentiation of osteoclasts.

Agents currently used as a treatment for osteoporosis include estrogen, latoxifene, alendronate and bisphosphonate, and these are used as bone resorption inhibitors to reduce the risk of fracture. In addition to such conventional therapeutic agents, studies on osteoporosis prevention, improvement, or therapeutic agents using calcium, vitamin D, calcitonin, biophosphonates, androgens and flavonoids are being actively conducted. However, these treatments have side effects to the human body, and there is a limit to being called a fundamental treatment for osteoporosis. Therefore, there is an urgent need for the prevention, improvement, or development of a therapeutic agent for osteoporosis derived from natural materials with few side effects while supplementing the disadvantages of existing therapeutic agents.

Vitex rotundifolia (Vitex rotundifolia) is a plant that grows along the sea breeze and is known as a halophyte that is resistant to salt. The flowers of the spruce tree are purple in color and have an excellent fragrance, and the fruits are used in folk remedies as a treatment for diseases such as headaches and eye diseases. Although research on some physiological activities such as skin beauty, antioxidant, anti-inflammatory, and antibacterial properties of S. serrata oleifera has been reported, no specific studies have been known on the prevention or treatment effect of Osteoporosis leaf extract.

Therefore, there is an urgent need for the development of a pharmaceutical composition for preventing or treating bone disease, and a food composition for preventing or improving bone disease, which include an extract of S.

Accordingly, the present inventors made intensive research efforts to develop a composition for improving, preventing or treating bone diseases. As a result, the present invention was completed by clarifying that the extract of Sunbigi tree exhibits an effect on the inhibition of osteoclast differentiation.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for preventing or treating bone disease, comprising an extract of S.

Another object of the present invention is to provide a food composition for improving bone disease, comprising an extract of S.

Another object of the present invention is to provide a method for preventing or treating bone disease, comprising administering to a subject in need thereof in an effective amount for preventing or treating bone disease of the extract of S.

Another object of the present invention relates to the improvement, prevention or treatment of bone disease of the extract of S.

The present invention relates to a pharmaceutical composition for preventing or treating bone disease, comprising an extract of Vitex rotundifolia as an active ingredient, and a food composition for improving bone disease.

Hereinafter, the present invention will be described in more detail.

One aspect of the present invention relates to a pharmaceutical composition for preventing or treating bone disease comprising an extract of Vitex rotundifolia as an active ingredient.

As used herein, the term "Sunbigi" is a tree of the Verbenaceae family, and it is a tree that grows in sand that is often seen on the shore. Also, it is a halophyte that grows with salt. It is a deciduous tree, and the leaves are thick and are opposite in ovate, obovate, and wide oval. The fruit is effective in antipyretic, sedative, and anti-inflammatory, so it is used as a treatment for cold sores, headache, arthritis, eye pain, customs pain, and tinnitus. The leaves and branches are fragrant and are used as spices.

In the present invention, Shunbigi tree may be one or more selected from the group consisting of leaves and stems, for example, may be leaves, but is not limited thereto.

As used herein, the term "extract" includes a solvent fraction of a crude solvent extract, a specific solvent soluble extract (solvent fraction), and a solvent fraction of a crude solvent extract.

In the present invention, the S. serrata extract may be an extract obtained by extraction with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms, but is not limited thereto.

In the present invention, the solvent has an alcohol content of 10% (v/v) to less than 100% (v/v), 20% (v/v) to 100% (v/v), 30% (v/v) v) greater than or equal to 100 % (v/v), greater than or equal to 40 % (v/v) and greater than or equal to 100 % (v/v), greater than or equal to 50 % (v/v) and less than or equal to 100 % (v/v), 50 % (v/v) or more and less than 95 % (v/v), 50 % (v/v) or more and less than 92 % (v/v), 60 % (v/v) or more to 100 % (v/v) ), less than 60% (v/v) to less than 95% (v/v), more than 60% (v/v) to less than 92% (v/v), more than 70% (v/v) to 100% (v/v) or more, 70 % (v/v) or more to 95 % (v/v), 70 % (v/v) or more to 92 % (v/v) or more, 80 % (v/) or more to less than 100% (v/v), 80% (v/v) to less than 95% (v/v), 80% (v/v) to less than 92% (v/v), e.g. It may be an aqueous alcohol solution of 90% (v/v), but is not limited thereto.

In the present invention, the solvent may be anhydrous alcohol, but is not limited thereto.

In the present invention, the solvent may be at least one selected from the group consisting of ethanol, butanol, ethyl ether and ethyl acetate, but is not limited thereto.

In the present invention, bone diseases include osteoporosis, periodontal disease, rheumatoid arthritis, multiple myeloma, degenerative arthritis, inflammatory alveolar bone resorption disease and inflammatory bone resorption disease. It may be at least one selected from the group consisting of, but is not limited thereto.

As used herein, the term “osteoporosis” refers to a condition in which bone strength is weakened due to a decrease in the amount of bone and a quality change, and thus a high possibility of fracture. Bone protects various organs in the human body and functions as a storage for substances necessary for the body such as calcium. After that, homeostasis is maintained. In the case of osteoporosis, as the two cell activities are out of balance, excessive bone destruction by osteoclasts occurs and progresses. A major molecule in osteoporosis is RANKL (receptoractivator of nuclear factor kappa-B ligand), which binds to its receptor and activates various transcription factors to promote osteoclast differentiation.

When RANKL is treated with macrophages, it is differentiated into TRAP-positive cells along with binding to RANK (Receptor activator of nuclear factor kappa-Β). When these TRAP-positive cells are stimulated with inflammatory factors such as RANKL and TNF-α, They fuse and differentiate into multinucleated TRAP-positive cells.

As used herein, the term "periodontal disease" refers to an inflammatory state of the dental support tissue caused by bacteria, and can be divided into gingivitis and periodontitis. The cause of the disease is the formation of a bacterial film when oral bacteria strikes due to poor oral hygiene. Dental plaque refers to a mass of bacteria that proliferates after bacteria attach to the tooth surface by using the sticky substance in saliva as an adhesive. If the dental plaque is left unattended, it can become inflamed and sometimes bleed from the gums and bad breath. These symptoms are called gingivitis. As gingivitis progresses further, the gap between the teeth and the gums becomes deeper, forming a periodontal pocket, where bacteria that cause periodontal disease multiply and periodontitis occurs. When periodontitis progresses, the gums bleed and swell even with mild stimulation such as brushing, and often turn into acute inflammation, causing pain. This inflammation lowers the bone-making function and increases the bone-resorbing action, causing the alveolar bone to gradually decrease, which leads to the destruction of the alveolar bone and eventually the loss of teeth.

As used herein, the term “arthritis” refers to a joint disease involving inflammation in one or more joint sites. A common form of the arthritis is osteoarthritis, which causes inflammation and pain due to damage to the bones and ligaments constituting the joint due to gradual damage or degenerative changes in the cartilage protecting the joint.

Another aspect of the present invention relates to a method for preparing an extract of Sunbigi tree.

The extract of Sunbigi tree includes a solvent extract of Sunbigi tree, as described above.

The manufacturing process of the Sunbigi extract according to the present invention will be described in more detail as follows: Extraction is performed at room temperature with an extraction solvent in an amount of about 10 to 50 times the weight of Sunbigi tree. After extraction, the filtrate is collected by filtration. The extraction temperature is not particularly limited, but for example, 15 to 110 °C, 20 to 100 °C, 20 to 90 °C, 20 to 70 °C, 20 to 50 °C, 20 to 30 °C, for example, 25 °C is good. .

The extraction process can be repeated once or several times, and in a preferred example of the present invention, a method of re-extraction after the primary extraction can be adopted, which is a method of extracting a large amount of its own moisture content even if it is effectively filtered in the case of mass production. This is to prevent this because the loss occurs because of high In addition, as a result of verifying the extraction efficiency for each step, it was found that about 80 to 90% of the total extraction amount was extracted by the secondary extraction.

In one example of the present invention, when the extraction process is repeated twice, the obtained residue is extracted again under reflux with an extraction solvent, about 5 to 15 times by volume, for example, 8 to 12 times by volume. After extraction, it is filtered, combined with the filtrate obtained previously, and concentrated under reduced pressure to prepare an extract of Sunbigi tree. In this way, extraction efficiency can be increased by mixing the filtrate obtained after the secondary extraction and each extraction, but the extract of the present invention is not limited to the number of extractions.

If the amount of the solvent used in the preparation of the extract is too small, stirring is difficult, the solubility of the extract is lowered, and the extraction efficiency is lowered. Since this may cause problems in handling, it is preferable to use the solvent within the above range.

The filtered extract thus obtained is about 10 to 30 times, for example 15 to 25 times, for example, about 20 times the total amount of the concentrate in order to adjust the content of the remaining lower alcohol so as to be suitable for use as a pharmaceutical raw material. After azeotropic concentration 1 to 5 times, for example, 2 to 3 times by weight with water, the same amount of water is added again to homogeneously suspend, and freeze-drying, it can be prepared as a powdery linden tree extract.

The extraction method used in the present invention may be any method commonly used, for example, hot water extraction, ultrasonic extraction, or reflux extraction may be used, but is not limited thereto.

The pharmaceutical composition of the present invention can be used as a pharmaceutical composition comprising a pharmaceutically effective amount of an extract of S.

As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to achieve the efficacy or activity of the above-mentioned Sunbigi extract.

Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. it is not going to be The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components.

The pharmaceutical composition according to the present invention may be administered to mammals including humans by various routes. The mode of administration may be any method commonly used, for example, may be administered by routes such as oral, skin, intravenous, intramuscular, subcutaneous, and the like, for example, orally.

A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and response sensitivity of the patient, An ordinarily skilled physician can readily determine and prescribe a dosage effective for the desired treatment or prophylaxis.

The composition of the present invention may further contain adjuvants such as pharmaceutically suitable and physiologically acceptable carriers, excipients, and diluents in addition to the mixed extract.

Carriers, excipients and diluents that may be included in the composition of the present invention 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 and mineral oil.

In the case of formulation, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be 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, for example, starch, calcium carbonate, sucrose ( sucrose) or lactose, gelatin, etc. may be mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Formulations for oral use include suspensions, solutions, emulsions, syrups, ointments, 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 include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, transdermal preparations, 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.

As the preparation of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, etc. can be used.

In an embodiment in which the composition of the present invention is applied to humans, the herbal extract composition of the present invention may be administered alone, but in general, it is combined with a pharmaceutical carrier selected in consideration of the administration mode and standard pharmaceutical practice. They may be mixed and administered.

For example, the composition containing the herbal extract of the present invention may be in the form of a tablet containing starch or lactose, or in the form of a capsule containing alone or an excipient, or an elixir or suspension containing a flavoring or coloring chemical. It can be administered orally, orally, or sublingually in the form of tablets. Such liquid formulations may contain suspending agents (eg, methylcellulose, semisynthetic glycerides such as witepsol or mixtures of apricot kernel oil and PEG-6 esters or PEG-8 and caprylic/capric glyceride mixtures, such as mixtures of glycerides).

The dosage of the extract-containing composition of the present invention may vary depending on the patient's age, weight, sex, dosage form, health status and disease level, and according to the judgment of the doctor or pharmacist, once to several times a day at regular time intervals It may be administered in divided doses.

In the present invention, the pharmaceutical composition contains 100 to 1000 mg/day, 100 to 900 mg/day, 100 to 800 mg/day, 150 to 1000 mg/day, 150 to 900 per 60 kg of body weight mg/day, 150 to 800 mg/day, 170 to 1000 mg/day, 170 to 900 mg/day, 170 to 800 mg/day, for example, 180 mg/day to 720 mg/day. it could be

If the daily dose of the composition containing the extract of S. serrata of the present invention is less than the above dose, a significant effect cannot be obtained. Since concerns may arise, it is preferable to set it within the above range.

Another aspect of the present invention relates to a food composition for improving bone disease comprising an extract of Vitex rotundifolia as an active ingredient.

In the present invention, Shunbigi tree may be one or more selected from the group consisting of leaves and stems, for example, may be leaves, but is not limited thereto.

In the present invention, the S. serrata extract may be an extract obtained by extraction with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms, but is not limited thereto.

In the present invention, the solvent has an alcohol content of 10% (v/v) to less than 100% (v/v), 20% (v/v) to 100% (v/v), 30% (v/v) v) greater than or equal to 100 % (v/v), greater than or equal to 40 % (v/v) and greater than or equal to 100 % (v/v), greater than or equal to 50 % (v/v) and less than or equal to 100 % (v/v), 50 % (v/v) or more and less than 95 % (v/v), 50 % (v/v) or more and less than 92 % (v/v), 60 % (v/v) or more to 100 % (v/v) ), less than 60% (v/v) to less than 95% (v/v), more than 60% (v/v) to less than 92% (v/v), more than 70% (v/v) to 100% (v/v) or more, 70 % (v/v) or more to 95 % (v/v), 70 % (v/v) or more to 92 % (v/v) or more, 80 % (v/) or more to less than 100% (v/v), 80% (v/v) to less than 95% (v/v), 80% (v/v) to less than 92% (v/v), e.g. It may be an aqueous alcohol solution of 90% (v/v), but is not limited thereto.

In the present invention, the solvent may be anhydrous alcohol, but is not limited thereto.

In the present invention, the solvent may be at least one selected from the group consisting of ethanol, butanol, ethyl ether and ethyl acetate, but is not limited thereto.

In the present invention, bone diseases include osteoporosis, periodontal disease, rheumatoid arthritis, multiple myeloma, degenerative arthritis, inflammatory alveolar bone resorption disease and inflammatory bone resorption disease. It may be at least one selected from the group consisting of, but is not limited thereto.

When the food composition of the present invention is used as a food additive, the food composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. In general, in the production of food or beverage, the food composition of the present invention may be added in an amount of 15% by weight or less, for example, 10% by weight or less based on the raw material.

There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages, vitamin complexes, and the like, and includes all foods in a conventional sense.

The beverage may contain various flavoring agents or natural carbohydrates as additional ingredients. As the above-mentioned natural carbohydrates, monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame may be used. . The ratio of the natural carbohydrate may be appropriately determined by the selection of those skilled in the art.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol , a carbonation agent used in carbonated beverages, and the like. In addition, the food composition of the present invention may contain fruit for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives may also be appropriately selected by those skilled in the art.

In the food composition, content overlapping with the pharmaceutical composition is omitted in consideration of the complexity of the present specification.

The present invention relates to a pharmaceutical composition for preventing or treating bone disease, and a food composition for improving bone disease, comprising an extract of Vitex rotundifolia as an active ingredient. As it exhibits an excellent effect on inhibition, it can be used as a pharmaceutical composition or a food composition useful for improving, preventing or treating various bone diseases.

1 is a photograph and graph showing the osteoclast inhibitory effect of a leaf extract of Sunbigi tree according to an embodiment of the present invention.

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

Example 1. Preparation of ethanol extract of Sunbigi leaf

Sunbigi leaves were collected from the coast of Wando-gun, Jeollanam-do in June 2020. Specifically, 100 g of leaves of Sunbigi tree were put in 1 L of 90% (v/v) ethanol, and then pulverized with a homogenizer and extracted at room temperature for 24 hours. Then, ethanol extract was obtained by filtration with No. 2 qualitative filter paper (8 um, Whatman) filter paper. 90% (v/v) ethanol (0.5 L) was added to the obtained residue, and extraction was performed again in the same manner as above. The obtained ethanol extract was concentrated under reduced pressure at 45° C. using an aspirator (CCA-1110, Eyela, Tokyo, Japan) to prepare an ethanol extract (0.81 g) of Sunbigi leaf.

Example 2. Effect of inhibiting osteoclast differentiation

2-1. Isolation and culture of mouse bone marrow-derived macrophages (BMDMs)

Bone marrow was isolated from femurs of 8 to 12-week-old C57BL/6 mice. Then, 10% (v/v) fetal bovine serum, 1% (v/v) penicillin and 25 ng/ml of macrophage-colony stimulating factor (M-CSF) were added to the obtained bone marrow. Bone marrow-derived macrophages (Bone marrow _- derived macrophage, BMDMs) were obtained.

2-2. Verification of osteoclast differentiation inhibition

Bone marrow-derived macrophages (BMDMs) were grown in a medium supplemented with 10% FBS, 1% penicillin and M-CSF (Macrophage-colony stimulating factor), 2 x 10 ⁵ cells per well Inoculated at a density and incubated for 24 hours in a 12-well plate (12-well plate). After replacing with the medium of the same composition as above, 10, 12.5, 15 and 20 μg/ml of ethanol extract of Sunbigi leaf ethanol was treated for 2 hours. Then, RANKL (Receptor activator of nuclear factor kappa-Βligand) was treated and reacted for 24 hours.

Osteoclast differentiation was induced for 4 days in the same manner as described above. Then, a chromogenic substrate was added to (Tartrate resistance acid phosphatase, TRAP), a cytochemical labeling enzyme of osteoclasts, and the nucleus was stained. Then, the cells multinucleated to three or more nuclei were imaged and quantified, and are shown in Table 1 and FIG. 1 below.

Ethanol extract from spruce leaf Treatment concentration (μg/ml) Number of osteoclasts (average) RANKL 2672 10 1586 12.5 1288 15 859 20 208

As can be seen in FIG. 1 , it was confirmed that the differentiation of osteoclasts was increased when macrophages were treated with RANKL. On the other hand, it was confirmed that when macrophages were treated with the ethanol extract of S. S. leaf ethanol extract, osteoclasts induced from RANKL were inhibited in a concentration-dependent manner. This indicates that the ethanol extract of S. serrata leaf has an excellent effect on inhibiting osteoclasts.

Claims (12)

A pharmaceutical composition for preventing or treating bone disease, comprising an extract of Vitex rotundifolia as an active ingredient. In the present invention, the spruce tree is at least one selected from the group consisting of leaves and stems, a pharmaceutical composition for preventing or treating bone disease. The agent for preventing or treating bone disease according to claim 1, wherein the extract of S. serrata is an extract obtained by extraction with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms. academic composition. The pharmaceutical composition for preventing or treating bone disease according to claim 3, wherein the solvent is an aqueous alcohol solution or anhydrous alcohol having an alcohol content of 10% (v/v) or more to less than 100% (v/v). According to claim 3, wherein the solvent is at least one selected from the group consisting of ethanol, butanol, ethyl ether and ethyl acetate, a pharmaceutical composition for preventing or treating bone disease. According to claim 1, wherein the bone disease is osteoporosis, periodontal disease, rheumatoid arthritis, multiple myeloma, degenerative arthritis (degenerative arthritis), inflammatory alveolar bone resorption disease and inflammatory bone A pharmaceutical composition for preventing or treating bone disease, which is at least one selected from the group consisting of resorption diseases. A food composition for improving bone disease, comprising an extract of Vitex rotundifolia as an active ingredient. [Claim 8] The food composition for improving bone disease according to claim 7, wherein the spruce tree is at least one selected from the group consisting of leaves and stems. The food composition for improving bone disease according to claim 7, wherein the S. serrata extract is an extract obtained by extraction with one or more solvents selected from the group consisting of water and a linear or branched alcohol having 1 to 4 carbon atoms. The food composition for improving bone disease according to claim 9, wherein the solvent is an aqueous alcohol solution or anhydrous alcohol having an alcohol content of 10% (v/v) or more to less than 100% (v/v). The food composition for improving bone disease according to claim 9, wherein the solvent is at least one selected from the group consisting of ethanol, butanol, ethyl ether and ethyl acetate. According to claim 7, wherein the bone disease is osteoporosis, periodontal disease, rheumatoid arthritis, multiple myeloma, degenerative arthritis (degenerative arthritis), inflammatory alveolar resorption disease and inflammatory bone A food composition for improving bone disease, which is at least one selected from the group consisting of resorption diseases.

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