KR20190042872A - Pharmaceutical composition for use in preventing or treating osteoporosis containing extract of crown daisy as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating osteoporosis containing an extract of Glebionis coronaria as an active component. The pharmaceutical composition according to the present invention significantly inhibits the production of osteoclasts, thereby being able to be widely used as medicine or health food. In addition, since the pharmaceutical composition does not show cytotoxicity, it is possible to have few side effects even when taking the same for a long time.

Description

[0001] The present invention relates to a pharmaceutical composition for preventing or treating osteoporosis, which contains an extract of Aspergillus oryzae as an active ingredient,

The present invention relates to a pharmaceutical composition for preventing or treating osteoporosis, which comprises an extract of Aspergillus oryzae as an active ingredient.

Osteoporosis is a disease caused by the collapse of the balance between osteoblasts and osteoclasts, resulting in bone resorption exceeding bone formation. In the onset of osteoporosis, the lime of the bone tissue is reduced, and the density of the bone becomes thinner, thereby widening the bone marrow. As the condition progresses, the bones become weaker, so that even a small impact is liable to fracture. It is known that osteoporosis is caused by aging, lack of exercise, low body weight, smoking, low calcium diet, menopause, and ovarian resection.

Osteoclast is a large, multi-nucleated cell that destroys or absorbs bone tissue that has become unnecessary in the process of bone growth of a vertebrate animal. If the activity of the cell is too active, the density of the bone is low and causes diseases such as osteoporosis. In order to differentiate into osteoclasts, RANKL (Receptor activator of nuclear factor kappa-B ligand) is required and macrophage proliferative factor (M-CSF) is required for osteoclast proliferation.

Differentiation of osteoclasts begins when RANKL binds to its receptor RANK. When RANKL binds to RANK, adapter molecules such as tumor necrosis factor receptor-associated factor 6 (TRAF6) are activated. TRAF2, TRAF5, and TRAF6, known as tumor necrosis factor receptor-associated factor family members, activate transcription factors such as NF-kB and activation protein-1 (AP-1) required for osteoclast differentiation. When TRAF6 is activated by RANKL signaling for osteoclast differentiation, phosphatidylinositol phosphorylase 3-phosphorylase (PI3K), transactivating factor beta activating phosphorylase (TAK1), protein kinase B (Akt) / PKB and Jun (MAPKs), NF-kB, c-Fos, Fra-1, the cyclic AMP inducible gene (CREB), the N-terminal kinase (JNK), the extracellular signal regulated kinase (ERK) , NFATc1 are activated. This induction is caused by the c-Fos pathway through TRAF6-NF-kB and RANKL-RANK signals. And osteoclast-specific genes such as cathepsin K, tartrate-resistant acid phosphatase (TRAP), calcitonin receptor, and osteoclast-related receptor (OSCAR) are expressed. The process by which the osteoclasts attach to the bone and break down the bone is called a cytoskeleton reoragnization involving the formation of a filamentous actin ring and a sealing zone formed by attaching to the bone surface around the erosion space . These RANKL / RANK / TRAF6 axes and NFATc1 are essential for the differentiation into mature osteoclasts, and these signaling pathways are initiated by RANKL, which can be a target for bone disease drugs.

Most osteoporosis medicines currently on the market are estrogenic substances, which cause side effects such as cancer, gallstones and thrombosis when administered over a long period of time. Osteoporosis is a disease that can not be treated only by short-term administration of a drug, and long-term administration of the drug is indispensable. Therefore, it is necessary to develop a new drug having a drug efficacy sufficient to replace estrogen without the side effects mentioned above even when the drug is administered for a long time.

Accordingly, research and development of osteoporosis treating agents using a variety of natural substances such as Ogaki extract, Acanthopanax root extract, Puerariae Radix extract, and Gossam extract are continuously being carried out. For example, a natural extract of Kwangbaek has been studied and developed as a drug for prevention or treatment of osteoporosis by significantly reducing the expression of osteoclast differentiation-related genes and the expression of transcription factors that regulate their expression (Patent Document 1).

On the other hand, Chrysanthemum coronarium is a 1-2 year old grass belonging to Compositae , and its origin is Mediterranean coast and it is also called Chunkyu. It has no hair on the whole, has a unique fragrance, and its stem is 30-80cm high with its branches divided. Leaves are alternate, double-leafed haploid leaf. The leaves are thick, the bottom is half-wrapped around the stem, and the back has a little shine. Yellow or white flowers bloom in early summer, but they are strong in flavor. In Europe, they are planted in flowerpots and flower beds, so they are only used for ornamental purposes. They are not used for food as in the Orient. On the other hand, it is widely known in Korea, China, and Japan for edible purposes, and leaves and stems are also used as medicines.

In addition, Suksagat is a calcium-rich and vitamin-rich food. It has been used for gastrointestinal function strengthening agent and constipation treatment for a long time. However, the preventive or therapeutic effect of osteoporosis caused by the osteoclast inhibitory activity of the mugwort extract has not been specifically studied.

Korean Patent Registration No. 10-1242635

The present inventors have made efforts to develop a new pharmaceutical composition having excellent osteoporosis prevention and treatment efficacy and a small side effect even after long-term administration, and confirmed that the mugwort mushroom extract inhibits osteoclastogenesis without showing cytotoxicity, .

Accordingly, it is an object of the present invention to provide a pharmaceutical composition and a health functional food which are excellent in osteoporosis prevention and treatment efficacy and have little side effects even when taken for a long time.

In order to achieve the above object,

The present invention provides a pharmaceutical composition for preventing or treating osteoporosis, which comprises an alcohol extract of Aspergillus oryzae as an active ingredient.

The composition may be characterized by inhibiting osteoclast differentiation.

The alcohol may be a C ₁ -C ₄ alcohol.

The composition may be formulated in the form of powders, granules, tablets, hard capsules, soft capsules or injections.

Further, the present invention provides a health functional food for preventing or ameliorating osteoporosis, which comprises an alcohol extract of Aspergillus oryzae as an active ingredient.

The health functional food may be characterized by inhibiting osteoclast differentiation.

The alcohol may be a C ₁ -C ₄ alcohol.

The pharmaceutical composition for prevention or treatment of osteoporosis containing the alcohol extract of Aspergillus oryzae according to the present invention exhibits an effect of significantly inhibiting the production of osteoclasts and can be widely used as a medicine or a health food.

In addition, it does not show cytotoxicity, and there is an advantage in that side effects are small even when taken for a long time.

Fig. 1 shows the results of Experimental Example 1 (1) in which the cytotoxicity of the mugwort extract was confirmed.
FIG. 2 shows the result of Experimental Example 1 (2) in which the effect of suppressing the differentiation of oocysts extract into osteoclasts was confirmed using TRAP staining method.
FIG. 3 is a graph showing the osteoclast numbers when Rancuglass-induced osteoclast is added to the RANKL-induced osteoblast according to (2) of Experimental Example 1.

Hereinafter, the present invention will be described in detail.

The terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary meanings and the inventor may properly define the concept of the term to describe its invention in the best possible way It should be construed as meaning and concept consistent with the technical idea of the present invention.

Pharmaceutical composition for preventing or treating osteoporosis

In one aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of osteoporosis, which comprises an alcohol extract of Aspergillus oryzae as an active ingredient.

The alcoholic extract of Prunus persicae may inhibit osteoclast differentiation and may be useful as a pharmaceutical composition for preventing or treating osteoporosis.

The alcohol may be a C ₁ -C ₄ alcohol.

More specifically, the alcohol may be a C ₁ to C ₄ alcohol or a mixture thereof, preferably ethanol.

When ethanol is used as the extraction solvent, it can extract the active ingredient more effectively, reduce the complexity of the extracted compound, have low cost, low toxicity and easy to remove.

The extract of Alaska pollack can be obtained by mixing 0.1 to 20 g of the powdered mugwort powder with 100 ml of alcohol and extracting the mixture at a temperature of 20 to 60 ° C for 12 to 36 hours. A preferred method for preparing the alcoholic extract of Mulberry beard used in the present invention will be described in more detail below.

As used herein, the term osteoporosis refers to a state in which the amount of bone is reduced and the strength of the bone is weakened due to a qualitative change, and the prevention, improvement and treatment of osteoporosis includes all kinds of diseases caused by decreased bone density and bone loss .

The pharmaceutical composition according to the present invention can be administered orally or parenterally at the time of clinical administration and can be used in the form of a general pharmaceutical preparation.

The composition may be formulated in the form of powders, granules, tablets, hard capsules, soft capsules or injections.

More specifically, the pharmaceutical composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, external preparations, suppository sterilized injection solutions, But the present invention is not limited thereto.

In the case of formulation, it may be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one or more excipients such as starch, calcium carbonate, sucrose, lactose, gelatin, . In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.

Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions and syrups. Various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc. may be included in addition to water and liquid paraffin which are simple diluents commonly used have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, and the like.

The composition of the present invention may contain at least one active ingredient which exhibits the same or similar function in addition to the above extract.

A preferable dosage of the pharmaceutical composition of the present invention can be appropriately selected depending on the condition and the weight of the patient, the degree of symptoms, the drug form, the administration route and the period. It is preferable for the composition of the present invention to allow the active ingredient to be administered at 0.2 mg / kg to 200 mg / kg per day for optimal efficacy. The administration may be carried out once a day or divided into several doses, but is not limited thereto.

Health functional food for preventing or improving osteoporosis

In another aspect, the present invention relates to a health functional food for preventing or ameliorating osteoporosis, which comprises an alcohol extract of Prunus persicae as an active ingredient.

The health functional food may be characterized by inhibiting osteoclast differentiation.

The alcohol may be a C ₁ -C ₄ alcohol.

More specifically, the alcohol may be a C ₁ to C ₄ alcohol or a mixture thereof, preferably ethanol.

When ethanol is used as the extraction solvent, it can extract the active ingredient more effectively, reduce the complexity of the extracted compound, have low cost, low toxicity and easy to remove.

The extract of Alaska pollack can be obtained by mixing 0.1 to 20 g of the powdered mugwort powder with 100 ml of alcohol and extracting the mixture at a temperature of 20 to 60 ° C for 12 to 36 hours. A preferred method for preparing the alcoholic extract of Mulberry beard used in the present invention will be described in more detail below.

The health functional food of the present invention can be manufactured in various forms such as, but not limited to, various drinks, gum, tea, confectionery, vitamin complex, health supplement and the like.

In addition, the health functional food of the present invention includes a case where it is added to a health food for the purpose of improving osteoporosis, and there is no particular limitation on the kind of the food. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The preferred intake amount of the health functional food of the present invention varies depending on the condition and body weight of the recipient, the degree of symptoms, the type of food, and the period of consumption, and can be appropriately selected. It is preferable for the health functional food of the present invention that the active ingredient is ingested at 0.2 mg / kg to 200 mg / kg per day for optimal effect.

A method for preparing oolong tea extract for prevention or treatment of osteoporosis

According to the present invention,

(a) lyophilization is carried out to obtain a powder, followed by mixing 0.1 to 20 g of the powder of chrysanthemum chrysanthemum per 100 ml of alcohol; (b) agitating the mixture at a temperature of 20 to 60 DEG C for 12 to 36 hours to elute the active ingredient; And (c) removing the solid content from the extraction solution and collecting and concentrating only the supernatant.

Hereinafter, the method for preparing the mugwort mugwort extract according to the present invention will be described step by step.

First, (a) Lentinus edodes are lyophilized to obtain a powder, and 0.1 to 20 g of chestnut powder is mixed per 100 ml of alcohol.

The mugwort powder is obtained by freeze-drying the mugwort powder. The mugwort powder to be mixed with the alcohol preferably has an average particle size of 0.1 to 0.5 탆, more preferably 0.25 탆 or less. When the wolfberry powder having an average particle size of 0.1 to 0.5 占 퐉 is used, the effective ingredient can be effectively extracted even at a low temperature for a short time while minimizing destruction of the active ingredient for prevention of osteoporosis. On the other hand, in order to control the particle size of the wolfberry powder obtained by freeze-drying, the powder may be pulverized by a pulverizer or the powder may be passed through a sieve to select only particles of a desired size.

Alcohols used as extraction solvents include, but are not limited to, lower alcohols of 1 to 4 or polyhydric alcohols, or mixtures thereof. Preferably, the alcohol may be ethanol. When ethanol is used as the extraction solvent, it can extract the active ingredient more effectively, reduce the complexity of the extracted compound, have low cost, low toxicity and easy to remove.

The ratio of the extraction solvent to the wolfberry powder is preferably from 0.1 to 20 g of wolfberry powder per 100 ml of alcohol, and more preferably from about 5 to 15 g, especially about 10 g of wolfberry powder per 100 ml of alcohol.

Next, (b) the active ingredient of osteoporosis treatment is eluted from the mixture of the extracting solvent and the cowguth powder at a temperature of 20 to 60 DEG C for 12 to 36 hours. In this case, the extraction temperature is more preferably 20 to 30 ° C, and most preferably, it is extracted at about room temperature of about 25 ° C. If the extraction temperature is higher than the above range, the active ingredient may be destroyed. If the extraction temperature is low, the extraction time becomes longer and the economical efficiency is lowered. The extraction time may vary depending on the extraction temperature, but most preferably about 24 hours.

When the extraction is completed, (c) the solid content is removed from the extraction solution, and only the supernatant is collected and concentrated. Separation of the solids and the supernatant may be carried out, for example, using a filter paper, or by centrifugation.

In addition, the step (d) may further comprise lyophilizing the concentrate of step (c) to obtain a powder.

Hereinafter, the pharmaceutical composition for prevention or treatment of osteoporosis containing the mugwuga extract of the present invention as an active ingredient and the application as a health functional food will be described in detail in the following Preparation Examples and Preparation Examples. However, the following Preparation Examples and Preparation Examples are for the purpose of specifying the contents of the present invention, and the present invention is not limited by the Preparation Examples and Formulation Examples.

< Manufacturing example >

Manufacturing example  1. Preparation of Mugwort mugwort extract

The mugwort mugwort was dried and then pulverized using a pulverizer. At this time, the average particle size of the wormwood powder is 0.1 to 0.5 탆. 500 g of 70 vol.% Aqueous ethanol solution was added to 100 g of the chrysanthemum powder, and the mixture was extracted with stirring at 25 DEG C for 24 hours. Then, the solid matter was removed at 8,000 DEG C for 30 minutes using a centrifuge (Beckman, USA). The centrifuged supernatant was collected, concentrated under reduced pressure, and lyophilized to give a powder.

< Formulation example >

Formulation example  One: Sanje  Produce

Artemisia powder (Preparation Example 1) - 300 mg

Lactose ------------------------------ 100 mg

Talc ------------------------------ 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation example  2: Preparation of tablets

Artemisia prunus powder (Preparation Example 1) - 50 mg

Corn starch ------------------------------ 100 mg

Lactose ------------------------------ 100 mg

Magnesium stearate ----------------------------- 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation example  3: Preparation of capsules

Sosogae powder (Preparation Example 1) - 50 mg

Corn starch ------------------------------ 100 mg

Lactose ------------------------------ 100 mg

Magnesium stearate ----------------------------- 2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  4: Preparation of injection

Artemisia prunus powder (Preparation Example 1) - 50 mg

Sterile sterilized distilled water for injection ---------------------------

pH adjusting agent -----------------------------

(2 ml) per 1 ampoule according to the usual injection preparation method.

Formulation example  5: Liquid  Produce

Powder of Artemisiaceae (Preparation Example 1) - 100 mg

Ising Party ------------------------------ 10 g

Mannitol ------------------------------ 5 g

Purified water ------------------------------

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, And sterilized to prepare a liquid preparation.

Formulation example  6: Of health food  Produce

Sosugtha powder (Preparation Example 1) - 1000 mg

Vitamin mixture ------------------------------

Vitamin A Acetate -------------------------- 70 g

Vitamin E ------------------------------ 1.0 mg

Vitamin B1 ------------------------------ 0.13 mg

Vitamin B2 ------------------------------- 0.15 mg

Vitamin B6 ------------------------------ 0.5 mg

Vitamin B12 ------------------------------ 0.2 g

Vitamin C ------------------------------ 10 mg

Biotin ------------------------------- 10 μg

Nicotinic amide ------------------------------ 1.7 mg

Folic acid ---------------------------------- 50 g

Calcium pantothenate ------------------------------ 0.5 mg

Inorganic mixture -------------------------------

Ferrous sulfate ------------------------------ 1.75 mg

Zinc oxide ------------------------------ 0.82 mg

Magnesium carbonate ------------------------------ 25.3 mg

Potassium phosphate monohydrate 15 mg

Secondary phosphate knife - 55 ㎎

Potassium citrate -------------------------- 90 mg

Calcium carbonate -------------------------------- 100 mg

Magnesium Chloride ------------------------------- 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Formulation example  7: Manufacture of health drinks

Sosugtha powder (Preparation Example 1) - 1000 mg

Citric acid ------------------------------ 1000 mg

Oligosaccharides ---------------------------- 100 g

Plum concentrate ----------------------------- 2 g

Taurine ------------------------------ 1 g

Purified water was added to the whole to 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The resulting solution was filtered and sterilized in a sterilized 2 L container, &Lt; / RTI &gt; Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Hereinafter, the effects of the alcohol extract of Mugwortgilliae according to the present invention on the treatment and prevention of osteoporosis will be described in more detail.

< Experimental Example >

Experimental Example  1. Determination of the Effect of Mugwort Extract on Inhibition of Osteoclastogenesis

(1) Cytotoxicity test

To measure the effect of the composition according to the present invention on cell survival, RAW 264.7 monocyte cells (monocyte cells before osteoclast differentiation) derived from mice were injected with 5, 10, 20, 40, 100 μg / And the survival rate was measured after 48 hours.

As a result, it was confirmed that the mugwugae extract according to the present invention was not involved in cell survival at all (see FIG. 1).

(2) Confirmation test of osteoclast differentiation inhibition effect

Osteoclast differentiation was observed using TRAP (tartrate resistant acid phosphate) staining method to confirm the effect of the composition according to the present invention on osteoclast differentiation.

For this purpose, RANKL (receptor activator of nuclear factor kappa-B ligand) was added to RAW264.7 mononuclear cells and treated with 0, 10, 20, 40, 100 μg / ml of TRAP .

RAW264.7 monocyte cells are differentiated into multinuclear osteoclasts when bound to RANKL, and TRAP enzyme increases secretion during bone resorption, so that the activity of the TRAP enzyme can be measured to confirm osteoclast differentiation.

As a result, it was confirmed that RANKL-induced osteoclast production in RAW264.7 monocyte cells was inhibited by the mugwort extract (see Fig. 2).

Particularly, when treated with 40 μg / ml of the mugwort extract of the present invention, there was a statistically significant difference (p <0.05) between the group treated with RANKL alone (see FIG. 3).

Claims (7)

A pharmaceutical composition for preventing or treating osteoporosis, which comprises an alcohol extract of Aspergillus oryzae as an active ingredient.
The method according to claim 1,
Wherein said composition inhibits osteoclast differentiation.
3. The method according to claim 1 or 2,
Wherein the alcohol is a C ₁ to C ₄ alcohol.
3. The method according to claim 1 or 2,
Wherein said composition is formulated in the form of powders, granules, tablets, hard capsules, soft capsules or injections.
A health functional food for preventing or ameliorating osteoporosis, which comprises an alcoholic extract of Zucchini as an active ingredient.
6. The method of claim 5,
Wherein said health functional food inhibits osteoclast differentiation.
6. The method of claim 5,
Wherein the alcohol is a C ₁ to C ₄ alcohol.

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