KR20050110809A - A herbal mixture extract of pleurotus eryngii and acanthopanacis cortex and composition comprising the same for prevention and treatment of osteoporosis - Google Patents

Abstract

The present invention relates to a mixed herbal medicine extract of the Greater Mushroom and Ogapi, and a composition for preventing and treating osteoporosis using the same as an active ingredient.

The mixed herbal medicine extract of the present invention proliferates osteoblasts, effectively inhibits the formation and activity of osteoclasts, and thus can be widely used as medicines and health foods useful for the prevention and treatment of osteoporosis.

Description

Herbal extract of Pleurotus eryngii and Acanthopanacis Cortex and composition comprising the same for prevention and treatment of osteoporosis

The present invention relates to a mixed herbal medicine extract of the Greater Mushroom and Ogapi, and a composition for preventing and treating osteoporosis using the same as an active ingredient.

Osteoporosis is a disease caused by a weak balance between bone resorption and bone formation. It is caused by excessive bone resorption than bone formation. Osteoporosis is caused by a decrease in the amount of lime in bone tissue, resulting in thinning of the bone. The bone marrow cavity (骨髓 腔) is widened, and as the symptoms progress, the bones become weak, so even a small impact is likely to fracture. Bone tissue is a dynamic tissue that is formed by osteoblasts and constantly breaks down and absorbed by osteoclasts.

Osteoporosis is not a symptom itself but rather various fractures that are easily caused by bone weakness, especially femoral fractures or vertebral fractures, which limit long-term activities and lead to a healthy life. It is known to do. Bone mass is influenced by several factors, including genetics, nutrition, hormone changes, differences in exercise and lifestyle, and the causes of osteoporosis include old age, lack of exercise, low weight, smoking, low calcium diet, menopause, Ovarian ablation and the like are known. On the other hand, although there are individual differences, blacks have lower bone resorption levels than whites, resulting in higher bone mass. Usually bone mass is highest at 14-18 years of age and decreases by about 1% per year. Especially in women, bone reduction continues after 30 years of age, and when menopause reaches bone growth rapidly due to hormonal changes. In other words, estrogen concentration rapidly decreases during menopause, in which a large amount of B-lymphocytes are produced, as in the case of interleukin-7 (IL-7). pre-B cells) accumulate and increase the amount of IL-6 thereby increasing the activity of osteoclasts.

As described above, osteoporosis is unavoidable for elderly people, especially postmenopausal women, and as the population ages in developed countries, interest in osteoporosis and its therapeutics is gradually increasing.

It is also known that there is a market of about $ 130 billion related to the treatment of bone diseases worldwide and is expected to increase further. Therefore, many research institutes and pharmaceutical companies around the world are investing heavily in the development of bone disease treatments. Currently, the development of bone resorption inhibitors is actively progressing.

In the past, the study was mainly focused on bone minerals, namely, calcium and phosphorus metabolic abnormalities, but no significant progress was made in determining the pathogenesis.

Materials currently used for the treatment of osteoporosis include bisphosphonate preparations (alendronate, ethidronate), hormone preparations (raloxifene), vitamin D preparations, calcitonin preparations, calcium preparations, and the like. However, bisphosphonate preparations are poorly absorbed, difficult to take, and cause esophagitis. Hormonal preparations should be taken for a lifetime. Unclear, calcitonin preparations are expensive and difficult to administer, and calcium preparations have fewer side effects but are limited to preventive effects rather than treatment.

Osteoporosis cannot be treated with short-term administration of the drug, but long-term administration of the drug is essential. Therefore, there is a need for a new substance that does not have such side effects and has good efficacy when the drug is administered for a long time.

Pleurotus eryngii, on the other hand, is a parasitic fungi belonging to the genus Pleurotus eryngii and Pleurotus eryngii. It occurs in the subtropical steppe and is called the Boletus of the steppes or King oyster mushroom in Europe. have. According to Dongbogam, large locust mushroom is nontoxic, has a sweet taste, has a strong aroma, and is used as a good medicine when the hands and feet are weak and have no power or have a low back and knee pain by helping the stomach and intestines to function and promote the circulation of energy. Its ingredients are not yet fully understood, but are alkaline foods, lower cholesterol levels and anti-cancer activity. Rich in protein and vitamins, it is known to be effective in tonsillitis, mastitis, dehydration and various adult diseases. In particular, it is an alkaline food, and is widely regarded as a natural health food with a high anticancer effect in Japan.

Ogapi, Acanthopanacis Cortex, is a shrub plant belonging to the Araliaceae family, distributed only in the Far Hemisphere of the Northern Hemisphere, and is an endangered wild plant in South Korea. Types of ogapi include thorn ogapi, general ogapi, scented ogapi, and it is classified as a medicinal herb that has no toxicity and side effects in Chinese medicine. Chisanoside has a pharmacological function in the leaves, and in the roots there are not only Ogapi glycosides but also Gilling (Sylrgin) and coumarin glycosides. Ogapi contains acanthosides (Acanthoside B, D) and water-soluble polysaccharides that increase immunity. Ogapi is known to be spicy, bitter, and warm in nature, acting on the liver and nerves, eliminating customs, invigorating and purifying essence. In addition, it was used to show the auropathy (sickness caused by the weakness of the five intestines) and the seven statues (seven symptoms caused by the weakness of a man) and to use the legs poorly. It is known to help and improve energy, to clear the mind, to increase the willpower, to prevent the body from becoming light and old, and to cure the bad blood inside the body cleanly and cleanly. Pain in the lumbar spine, men's vulgaris, speech, women It is known to cure various symptoms such as yin and yang.

Thus, the inventors of the present invention, while conducting various physiological activity studies of the Great Oyster mushroom and Ogapi, the mixed herbal extract of the Great Oyster mushroom and Ogapi is not only toxic but also shows the activity to proliferate osteoblasts and inhibit the proliferation of osteoclasts The present invention has been completed by revealing its effectiveness in preventing and treating osteoporosis without side effects.

The present invention is to provide a mixed herbal medicine extract having osteoblast proliferation and osteoclast proliferation inhibitory activity mixed with the extract of Oyster mushroom.

In addition, the present invention is to provide a composition for preventing and treating osteoporosis using the mixed herbal medicine extract as an active ingredient.

The present invention provides a mixed herbal medicine extract having osteoblast proliferation and osteoclast proliferation inhibitory activity mixed with the extract of Oyster mushroom.

The present invention also provides a composition for the prevention and treatment of osteoporosis using the mixed herbal medicine extract as an active ingredient.

Compositions of the present invention include pharmaceutical and health food compositions useful for the prevention and treatment of osteoporosis.

Hereinafter, the present invention will be described in detail.

The present invention provides a herbal extracts mixed with 0.5 ~ 2.5 parts by weight of the extract of Oyster Mushroom and 0.5 ~ 5.0 parts by weight of the Ogapi extract.

The mixed herbal medicine included in the composition of the present invention is used by extracting with water or alcohol, and the extraction method of the mixed herbal medicine is as follows.

Put a large mushroom and Ogapi into the extraction container, and distilled water was added to extract hot water at 100 ℃ for 4 hours. After repeating the above procedure three times, the resulting solution is cooled to room temperature and filtered through a filter paper. Each of the extracts obtained was concentrated under reduced pressure at 40 ° C. or lower using a vacuum rotary evaporator and lyophilized to obtain a powder of Oyster mushroom extract and Ogapi extract, respectively. To prepare a mixed herbal medicine extract by mixing 0.5 to 2.5 parts by weight and 0.5 to 5.0 parts by weight of the extract of the large locust mushroom obtained from the above.

The present invention also provides a composition for the prevention and treatment of osteoporosis using the mixed herbal medicine extract as an active ingredient.

The mixed herbal medicine extract of the present invention exhibits a cell proliferation effect of about 120-130% or more as compared to the case of treating the Greater Black Mushroom Extract and the Ogapi Extract, respectively. Increase it by ~ 150%.

In addition, the mixed herbal medicine extract of the present invention significantly inhibits the production and activity of osteoclasts as compared to the case of treating the Greater Mushroom extract and the Ogapi extract alone.

Therefore, the mixed herbal medicine extract of the present invention exhibits excellent activity on basic phosphatase, which is a marker enzyme of osteoblasts, is effective in osteoblast proliferation, and significantly inhibits the production and activity of osteoclasts, thus effectively preventing or treating osteoporosis. Can be used.

The composition of the present invention may further contain at least one active ingredient exhibiting the same or similar function in addition to the mixed herbal medicine extract.

The mixed herbal extracts can be administered orally or parenterally during clinical administration and can be used in the form of general pharmaceutical formulations. That is, the mixed herbal extract of the present invention may be administered in various oral and parenteral formulations during actual clinical administration, and when formulated, diluents such as fillers, extenders, binders, humectants, disintegrating agents, and surfactants are usually used. Or using excipients. Solid preparations for oral administration include tablets, pills, powders, granules and capsules, and the like, which may be used in mixed herbal extracts, including at least one excipient such as starch, calcium carbonate, sucrose, lactose and gelatin, and the like. Are mixed to prepare. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups, and may include various excipients such as wetting agents, sweeteners, fragrances and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol and gelatin may be used.

Dosage varies depending on the patient's weight, age, sex, health condition, diet, time of administration, method of administration, excretion rate and severity of the disease. The daily dosage is 150 to 450 mg / kg mixed herbal medicine extract. It is preferably 300 to 400 mg / kg, it may be administered 1 to 6 times a day.

As a result of conducting toxicity experiments by orally administering the mixed herbal extract of the present invention to rats, the 50% lethal dose (LD ₅₀ ) by the oral administration toxicity test is determined to be a safe substance of at least 1 g / kg or more.

The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of osteoporosis.

The composition of the present invention may be added to health food for the purpose of improving osteoporosis. When the mixed herbal medicine extract of the present invention is used as a food additive, the mixed herbal medicine extract may be added as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). In general, in the preparation of food or beverage, the mixed herbal extract of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less based on the raw material. However, in the case of long-term intake for health and hygiene or health control, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety. .

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

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the general beverage. The above-mentioned natural carbohydrates are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonic acid. Carbonating agents and the like used in beverages. In addition, the composition of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not critical but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, preferred examples and experimental examples are presented to help understand the present invention. However, the following Examples and Experimental Examples are provided only to more easily understand the present invention, and the contents of the present invention are not limited by the Examples.

Example  : Preparation of Mixed Herbal Extracts

1. Preparation of Greater Mushroom Extract

100 g of Greater Mushroom was placed in a 3 L extraction vessel, and 1 L of distilled water was added, followed by hot water extraction at 100 ° C. for 4 hours. After repeating the above procedure three times, the resulting solution was cooled to room temperature and filtered through a filter paper. The extract was concentrated under reduced pressure at 40 ° C. or lower using a vacuum rotary evaporator and lyophilized to obtain a powder of the large nectar mushroom (yield: 42%).

2. Preparation of Ogapi Extract

100 g of Ogapi was placed in a 3 L extraction vessel, and 1 L of distilled water was added, followed by hot water extraction at 100 ° C. for 4 hours. After repeating the above procedure three times, the resulting solution was cooled to room temperature and filtered through a filter paper. The extract was concentrated under reduced pressure at 40 ° C. or lower using a vacuum rotary evaporator and lyophilized to obtain an extract of Ogapi powder (yield: 5.8%).

3. Preparation of Mixed Herbal Extracts

0.5 to 2.5 parts by weight of the extract of the large locust mushroom obtained in step 1 and 0.5 to 5.0 parts by weight of the extract of the ogapi obtained in step 2 were prepared in a mixed herbal medicine extract.

The mixed herbal extracts were diluted in the culture and used for the following experiments.

Experimental Example 1  : Analysis of Osteoblast Proliferation Efficacy by Mixed Herbal Medicine Extracts of the Present Invention

In order to determine the effect of the mixed herbal medicine extract of the present invention on osteoblast proliferation and cell activity, the following experiment was performed.

1. Osteoblast proliferation and cell activity analysis

In order to evaluate the effect of the mixed herbal medicine extract of the present invention on the proliferation of osteoblasts, MG-63 and HOS cell lines derived from human osteosarcoma were purchased from ATCC (American Type Culture Collection, Rockville, USA) and used. Cultured in Dulbecco's modified Eagle's medium (DMEM) containing% FBS (fetal bovine serum) was used.

Indirectly evaluating whether the mixed herbal medicine extract of the present invention is toxic, and also to determine the effect of the mixed herbal medicine extract on the proliferation of osteoblasts, MG-63 osteoblasts cultured in The amount of ³ H-thymidine introduced into the DNA was measured. Specifically, osteoblasts were dispensed into 24-well plates to be 2 × 10 ⁴ cells per well and replaced with DMEM containing 1% FBS the next day. In the replaced culture, the mixed herbal extract prepared in the above example was diluted to a concentration of 1 × 10 ⁻⁴ to 1 mg / ml, added at different concentrations, and incubated for 48 hours. After incubation with 3 μCi / well of ³ H-cymidine for the last 4 hours, the cells were washed with PBS (phosphate buffered saline) buffer and treated with 5% trichloroacetic acid (TCA) at 4 ° C. Fractions were separated and dissolved in 0.1 M NaOH. The influx of ³ H-cymidine was measured by taking a certain fraction of these and measuring the radioactivity at the liquid scintillation counter.

The results are shown in Table 1 and FIG.

Concentration (mg / ml) Ogapi Extract 0 0.5 1.0 5.0 10.0 Black Oyster Mushroom Extract 0 100.0 105.2 109.9 112.5 119.1 0.5 108.9 109.1 112.2 115.8 121.3 1.0 112.9 112.5 123.8 126.5 132.1 5.0 118.2 119.8 129.2 132.7 138.1 10.0 120.2 126.8 132.5 135.2 145.8

As shown in Table 1 and Figure 1, the cells added with the mixed herbal medicine extract of the present invention showed a cell proliferation effect of about 120 ~ 130% or more compared to the cells added alone with the extract of Oyster mushroom and Ogapi extract.

2. Measurement of basic phosphatase activity, osteoblast marker

In order to evaluate the effect of the mixed herbal medicine extract of the present invention on the activity of osteoblasts, the HOS osteoblasts cultured in 1 above were dispensed in 96-well plates to be 5 × 10 ³ cells per well, and grown in monolayers. The mixed herbal extracts were then added to the culture at concentrations of 0.08, 0.4, 1.0, 5.0, 10.0 mg / ml, and cultured for 48 hours in 96-well plates with DMEM containing 10% FBS. Treated with 100 / saline for 30 minutes at 37 ° C. PNP (p-nitrophenyl) liberated from the substrate PNPP by reacting a certain amount of the cell treatment solution with 0.1N glycine-NaOH buffer (pH 10.4) for 30 minutes at 37 ° C in the presence of 100 mM PNPP (p-nitrophenylphosphate) as a substrate. ) Was colorimetrically determined at 405 nm.

The results are shown in Table 2 and FIG.

Concentration (mg / ml) Ogapi Extract 0 0.5 1.0 5.0 10.0 Black Oyster Mushroom Extract 0 100.0 114.0 115.0 118.0 121.0 0.5 114.0 116.0 120.0 120.0 130.0 1.0 122.0 136.9 139.2 141.1 145.2 5.0 128.0 141.3 145.8 148.5 155.4 10.0 132.0 146.8 151.8 155.7 158.9

As shown in Table 2 and Figure 2, the cells with the mixed herbal medicine extract of the present invention increased the activity of basic phosphatase by about 110-150% compared to the cells added with the extract of Oyster mushroom and Ogapi extract alone. .

Therefore, the mixed herbal medicine extract of the present invention exhibits excellent activity on basic phosphatase, which is a labeling enzyme of osteoblasts, and thus has an effect on osteoblast proliferation.

Experimental Example 2  : Proliferation inhibitory effect of osteoclasts (osteoclast) by the mixed herbal medicine extract of the present invention

In order to examine how the mixed herbal medicine extract of the present invention affects the growth and activity of osteoclasts, osteoclast progenitor cells were cultured on calcium-phosphate-coated plates (OAAS, OCT Inc.) TRAP (Tartrate-resistant acid phosphatase; hereinafter abbreviated as "TRAP") activity was analyzed.

1. Pure separation of osteoclast progenitor cells and differentiation into mature osteoclasts

In order to separate the mouse bone marrow cells, 7 to 9 weeks old male mice were sacrificed with cervical torsion, followed by sterile extraction of the femur and tibia, removal of soft tissues, and the cutting of both ends of the iliac bone. 1 ml of enzyme solution containing 0.1% collagenase (Gibco), 0.05% trypsin and 0.5 mM EDTA (Gibco) was injected into the bone marrow cavity to remove the bone marrow, stirred for 30 minutes, and the bone marrow cells were collected to contain 10% FBS. After incubation in α-MEM (α-minimum essential medium) for 24 hours, unattached cells were collected. Unattached cells, which become progenitor cells of osteoclasts, were cultured by aliquoting 2 × 10 ⁵ cells per well. Mixed herbal medicine of the above example in α-MEM containing 20ng / ml macrophage-colony stimulating factor (M-CSF, Peprotech, USA) and 30ng / ml RANKL (Peprotech, USA) during 8 days of culture The extract was treated and incubated. After incubation, the cells were fixed and subjected to TRAP staining to examine the production of osteoclasts.

2. Measurement of Multinucleated Cell Formation as TRAP (+)

The measurement of osteoclast formation was observed by the number of multinucleated cells showing TRAP (+) after culturing osteoclast progenitor cells on a plate coated with calcium-phosphate.

Specifically, after the cell culture, the adherent cells were washed with PBS and then fixed with citrate-acetate-formaldehyde for 5 minutes, naphthol AS-BI phosphate, fast garnet ( fast Garnet) TRAP staining by incubating for 1 hour in 37 ℃ acetate buffer (pH 5.0) containing GBC solution and 7mM tartrate buffer (pH 5). TRAP (+) multinucleated cells with three or more nuclei were considered as osteoclasts.

The results are shown in Table 3 and FIG.

Osteoclast proliferation inhibition rate (%) Concentration (mg / ml) Ogapi Extract 0 0.5 1.0 2.5 5.0 Black Oyster Mushroom Extract 0 0 14.77 61.08 92.22 99.20 0.5 16.57 53.29 77.64 98.00 100 1.0 31.54 74.45 95.21 98.80 100 2.5 87.03 99.80 100 100 100

As shown in Table 3 and FIG. 3, the control ratio of control group was 0 when treated with the Great Pleurotus eryngii extract and the inhibition rate of the number of TRAP positive multinucleated cells compared to the control group was 0.5, 1.0, 2.5 mg / ml, respectively. At the concentration of 16.57, 31.54, 87.03% inhibited the production and activity of osteoclasts. In addition, when the Ogapi extract was treated alone, the formation and activity of osteoclasts were inhibited by 14.77, 61.08, 92.22, and 99.20% at concentrations of 0.5, 1.0, 2.5, and 5.0 mg / ml, respectively.

In addition, when the concentrations of the extract of Oyster mushroom and Ogapi extract were mixed at 0.5 mg / ml or more, respectively, the production and activity of the osteoclasts was significantly inhibited compared to the case of treatment alone.

Therefore, it can be seen that the mixed herbal medicine extract of the present invention significantly inhibits the formation and activity of osteoclasts as compared to the case of treating the Greater Mushroom extract and the Ogapi extract alone.

Experimental Example 3  : Acute Toxicity of Oral Administration in Rats

In order to determine the acute toxicity of the mixed herbal medicine extract of the present invention, an acute toxicity test was carried out by the following method.

Acute toxicity test was conducted using 6-week-old specific pathogen-free (SPF) SD rats. Two animals per group were suspended orally administered once with a dose of 1 g / kg / ml in suspension of the mixed herbal medicine of the present invention in 0.5% methylcellulose solution, respectively. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and hematological examinations were performed. Necropsy was performed to observe abdominal and thoracic organ abnormalities.

As a result, no significant clinical symptoms or dead animals were noted in all animals treated with the test substance, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, and autopsy findings.

Therefore, the mixed herbal medicine extracts of the present invention do not show any toxic change even in rats up to 1 g / kg, and the minimum lethal dose (LD ₅₀ ) of oral administration was determined to be a safe substance of more than 1 g / kg of extract.

Examples of preparations for the compositions of the present invention are illustrated below.

Formulation Example 1  : Preparation of Pharmaceutical Formulations

1. Preparation of powder

Mixed Herbal Extract 2g

1g lactose

The above ingredients were mixed and filled in airtight cloth to prepare a powder.

2. Preparation of Tablets

Mixed Herbal Extract 100mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

3. Preparation of Capsule

Mixed Herbal Extract 100mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

Formulation Example 2  : Manufacture of food

Food products containing the mixed herbal medicine extract of the present invention were prepared as follows.

1. Preparation of Cooking Seasonings

20 to 95% by weight of the mixed herbal medicine extract of the present invention was prepared for cooking spices for health promotion.

2. Preparation of Tomato Ketchup and Sauce

0.2 ~ 1.0% by weight of the mixed herbal medicine extract of the present invention was added to tomato ketchup or sauce to prepare tomato ketchup or sauce for health promotion.

3. Manufacturing of Flour Foods

0.5 ~ 5.0% by weight of the mixed herbal medicine extract of the present invention was added to the flour, and using the mixture to prepare bread, cakes, cookies, crackers and noodles to prepare a health promoting food.

4. Preparation of soups and gravy

0.1 ~ 5.0% by weight of the mixed herbal medicine extract of the present invention was added to soups and broth to prepare meat products for health promotion, soups and noodles of noodles.

5. Preparation of Ground Beef

10% by weight of the mixed herbal medicine extract of the present invention was added to ground beef to prepare ground beef for health promotion.

6. Manufacture of Dairy Products

5 ~ 10% by weight of the mixed herbal medicine extract of the present invention was added to the milk, using the milk to prepare a variety of dairy products such as butter and ice cream.

7. Manufacture of wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh using a grinder.

Black beans, black sesame seeds, and perilla were also steamed and dried by a known method, and then ground to a powder having a particle size of 60 mesh.

The mixed herbal medicine extract of the present invention was decompressed and concentrated in a vacuum concentrator, and the dried product obtained by drying with a spray and a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

It was prepared by combining the dry powder of the grains, seeds and mixed herbal medicines prepared in the following ratio.

Cereals (30% by weight brown rice, 15% by weight barley, 20% by weight barley),

Seeds (7% by weight perilla, 8% by weight black beans, 7% by weight black sesame),

Dry powder (3% by weight) of the mixed herbal medicine extract,

Ganoderma lucidum (0.5% by weight),

Sulfur (0.5 wt%)

Formulation Example 3  : Preparation of Beverages

1. Preparation of Carbonated Drinks

5-10% of sugar, 0.05-0.3% citric acid, 0.005-0.02% caramel, 0.1-1% of vitamin C are mixed and 79-94% purified water is mixed to make syrup, and the syrup is 85-98 Sterilizing for 20 ~ 180 seconds at ℃ and mixed with a cooling water in a ratio of 1: 4 and then injected with 0.5 ~ 0.82% carbon dioxide gas to prepare a carbonated beverage containing the mixed herbal medicine extract of the present invention.

2. Manufacture of health drinks

Instant sterilization by homogeneously mixing subsidiary ingredients such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), water (75%) and mixed herbal extracts , Packed in a small packaging container such as plastic bottles to prepare a healthy beverage.

3. Preparation of Vegetable Juice

5 g of the mixed herbal medicine extract of the present invention was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion.

4. Preparation of Fruit Juice

1 g of the mixed herbal medicine extract of the present invention was added to 1,000 ml of apple or grape juice to prepare fruit juice for health promotion.

The mixed herbal medicine extract of the present invention proliferates osteoblasts, effectively inhibits the formation and activity of osteoclasts, and thus can be widely used as medicines and health foods useful for the prevention and treatment of osteoporosis.

1 is a diagram showing the effect of the mixed herbal medicine extract of the present invention on osteoblast proliferation.

Figure 2 is a diagram showing the effect of the mixed herbal medicine extract of the present invention on the activity of basic phosphatase of osteoblasts.

Figure 3 is a diagram showing the inhibition of osteoclast proliferation by the mixed herbal medicine extract of the present invention.

Claims (5)

Mixed herbal medicine extracts having osteoblast proliferation and osteoclast proliferation inhibitory activity in combination with the extract of Oyster mushroom and Ogapi extract. According to claim 1, Mixed herbal medicine extract, characterized in that the 0.5 to 2.5 parts by weight of the extract of the large locust mushroom and 0.5 to 5.0 parts by weight of the Ogapi extract. A therapeutic agent for osteoporosis comprising the mixed herbal extract of claim 1 or 2 as an active ingredient. A pharmaceutical composition for preventing and treating osteoporosis, comprising the mixed herbal extract of claim 1 or 2 as an active ingredient. Claim 1 or claim 2, wherein the mixed herbal medicine extract as an active ingredient osteoporosis prevention and treatment health food composition.