KR101914202B1 - Composition for treating, preventing or alleviating bone diseases comprising extract of edgeworthia papyrifera - Google Patents

Abstract

The present invention relates to a composition for treating, preventing or alleviating bone diseases comprising an extract of Edgeworthia papyrifera as an effective component. The composition of the present invention promotes the generation of bones and suppresses the absorption of the bones at the same time, thereby providing fundamental treatment, prevention, and alleviation effects for bone disease.

Description

TECHNICAL FIELD The present invention relates to a composition for treating, preventing or ameliorating osteoporosis, comprising a extract of Tricholoma matsutake mushroom,

The present invention relates to a pharmaceutical composition for preventing or treating osteoporosis and an osteoporosis-improving food composition comprising an extract of Aspergillus oryzae as an active ingredient.

Normal human bone maintains homeostasis by balancing bone formation by osteoblasts and osteoclast-induced bone resorption. However, when the homeostasis of osteogenesis and bone resorption is not regulated and the osteoclast differentiation, formation and activity are excessively increased, diseases resulting in bone tissue disorders or bone tissue destruction may be induced. Specifically, it is known to cause osteoporosis, hypercalcemia and bone metastasis to some cancers, or increase bone loss to rheumatoid arthritis.

In particular, osteoporosis is a disease in which the balance between osteoblasts and osteoclasts collapses and bone resorption precedes bone formation. Patients with osteoporosis are more likely to have active bone resorption than bone formation, resulting in increased bone loss, resulting in reduced bone mass, reduced bone mass, and decreased bone strength. In order to effectively treat or prevent osteoporosis, it is necessary to inhibit the activity of osteoclasts or to increase the activity of osteoblasts. Presently, drugs used as therapeutic agents for osteoporosis include bisphosphonate preparations, hormone preparations, vitamin D preparations, calcitonin preparations, and calcium preparations. However, most of the currently applied drugs have a limited effect on osteogenesis as a bone resorption inhibitor, which has limitations in preventive and therapeutic effects, and adverse effects such as gastrointestinal disorders and breast cancer induced by long-term administration have been reported. In addition, even a small number of osteogenesis promoters are limited in their use due to side effects such as hypocalcemia and high cost due to protein and peptide-based drugs.

Thus, in addition to fundamental and new research for the treatment of diseases associated with bone loss including osteoporosis, it is possible to control bone homeostasis by simultaneously controlling bone resorption and promoting bone formation, thereby preventing or restoring bone loss Development of material is urgent. In addition, osteoporosis including osteoporosis is essential for long-term administration of drugs, so it is urgently required to develop new medicines having low side effects and safe and excellent pharmaceutical efficacy.

Therefore, the present inventors have made intensive efforts to develop a material that promotes bone formation and inhibits bone uptake. As a result, it has been found that the extract of Tricholoma matsutake extract promotes the differentiation and activity of osteoblasts and the differentiation of osteoclasts into mature osteoclasts And the present invention has been completed.

Chinese Patent Laid-Open Publication No. CN 103977182 A (Apr. 13, 2013)

It is an object of the present invention to provide a pharmaceutical composition for treating or preventing bone diseases.

It is another object of the present invention to provide a food composition for improving or preventing bone diseases.

In order to accomplish the above object, one aspect of the present invention provides a pharmaceutical composition for treating or preventing bone diseases, comprising an extract of Tricholoma matsutake mushroom as an active ingredient.

The term " Edgeworthia papyrifera " as used in the present invention is a broad-leaved shrub, which is mainly used for textiles or ornamental purposes. In the composition according to the present invention, cultivated, harvested or commercially available bean curd refuse can be used without limitation.

As used herein, the term " bamboo shoot extract " means an extract obtained from bamboo shoot tree. The extract of Tricholoma matsutake mushroom contained in the pharmaceutical composition of the present invention may be one obtained by extracting at least one member selected from the group consisting of roots, stems, branches, flowers, leaves and fruits of Tricholoma matsutake. Preferably, it may be extracted from the stem of the leaves.

In addition, the extract of Tricholoma matsutake mushroom contained in the composition of the present invention may be obtained by extracting Tricholoma matsutake with a solvent selected from the group consisting of water, an organic solvent and a mixture thereof. The organic solvent may be, for example, one or more solvents selected from the group consisting of lower alcohols having 1 to 4 carbon atoms, chloroform, ethyl acetate, hexane, dichloromethane, acetone and diethyl ether. The solvent for obtaining the extract of Tricholoma matsutake can be preferably an organic solvent, more preferably methanol, ethyl acetate, or a mixture thereof.

The term " extract " used in the present invention means a preparation obtained by extracting a herbal medicine with an appropriate extraction solvent and evaporating the extraction solvent. The term " extract " means a product obtained by extracting a diluted solution, , A controlled-release product thereof, or a purified product thereof. For example, the extract of hot water extract, hot water extraction, cold extraction, reflux cooling, ultrasonic extraction, etc. can be used for the extract of Tricholoma matsuri according to the present invention.

The pharmaceutical composition according to the present invention may comprise a compound represented by the following general formula (I) or a pharmaceutically acceptable salt thereof,

[Chemical Formula 1]

.

.

The compound of formula (1) is edgeworoside A.

The pharmaceutically acceptable salts of the compounds of formula 1 above represent any and all organic or inorganic addition salts of the compounds which may be prepared by conventional methods in the art and include, for example, hydrochloric acid, hydrogen bromide And salts with inorganic acids such as hydrochloric acid, sulfuric acid, sodium hydrogen sulphate, phosphoric acid and carbonic acid or salts with inorganic acids such as formic acid, acetic acid, oxalic acid, benzoic acid, citric acid, tartaric acid, gluconic acid, gestic acid, fumaric acid, lactobionic acid, salicylic acid, Together with an organic acid such as acetic acid (aspirin), to form a pharmaceutically acceptable salt of these acids, or to react with an alkali metal ion such as sodium or potassium to form a metal salt thereof, or to react with an ammonium ion to form another Lt; RTI ID = 0.0 > pharmaceutically < / RTI > acceptable salt.

The pharmaceutical composition according to the present invention can prevent or treat bone diseases. As used herein, the term " bone disease " includes conditions caused by loss of activity of osteoblasts or excessive production and / or migration of osteoclasts, or diseases and diseases of lowering bone mass. The term "lowering bone disease" refers to a condition or disease in which a decrease in bone mass accompanied by a symptom such as a decrease in bone density and a degradation of bone tissue occurs, which is caused by osteoporosis caused by estrogen depression in menopausal age, Bone loss due to osteoclasts may also be included in the scope of the present invention. The bone disease is preferably selected from the group consisting of fractures, osteoprosis, osteomalacia, osteopenia, osteoarthritis, rheumatoid arthritis, periodontal disease, metastatic bone cancers, And Paget disease, and more preferably, it may be osteoporosis.

As used herein, the term " treatment " refers to any action that improves or alleviates the symptoms of bone disease by administration of the composition of the present invention. As used herein, the term " prevention " means any action that inhibits or delays the symptoms of bone disease by administration of the composition of the present invention.

The content of the extract of Tricholoma matsutake mushroom extract in the pharmaceutical composition of the present invention can be appropriately adjusted according to the symptoms of the disease, the progress of symptoms, the condition of the patient, and the like. For example, from 0.0001 to 99.9% by weight, from 0.1 to 90% by weight, from 1 to 80% by weight, from 1 to 70% by weight, from 1 to 60% by weight, or from 1 to 50% by weight, based on the total weight of the composition.

The pharmaceutical composition of the present invention may further comprise an additional component in addition to the above-mentioned extract of Samburu mallow extract for enhancing the efficacy. The additional ingredients may be, for example, minerals such as calcium, phosphorus, vitamin D, or a combination thereof, preferably calcium, phosphorus, or combinations thereof.

The pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. Examples of carriers, excipients and diluents that may be contained in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, But are not limited to, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral.

The pharmaceutical composition of the present invention may preferably be an oral dosage form. In addition, the pharmaceutical compositions according to the present invention may be formulated into suitable dosage forms for topical administration using techniques well known to those skilled in the art, and in the case of topical dosage forms, the formulations may include external preparations, foams, suppositories, have. In one embodiment, the pharmaceutical composition of the present invention may be formulated as an external preparation by mixing the extract with a base, which is well known in the art and commonly used. The external preparation may contain emulsions, gels, ointments, creams, patches, liniments, powders, aerosols, sprays, lotions, serums, pastes, foams, drops, suspensions and tinctures.

The pharmaceutical composition of the present invention can be formulated in the form of oral dosage forms such as pills, powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method have. In the case of formulation, it can 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 excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, Can be mixed and prepared. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories.

The pharmaceutical composition of the present invention is administered to a subject in a pharmaceutically effective amount. As used herein, the term " pharmaceutically effective amount " means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment for medical treatment, and the effective dose level will depend on the species and severity, Sex, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of excretion, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art.

The pharmaceutical composition of the present invention may be varied depending on the patient's age, sex, and body weight. Specifically, the composition of the present invention may be administered at a dose of 0.1 to 100 mg / kg once or several times a day depending on the degree of bone loss of a bone disease patient . In addition, the dose may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, bone loss, Accordingly, the dosage is not limited in any way to the scope of the present invention.

Yet another aspect of the present invention provides a food composition for improving or preventing bone diseases, which comprises the extract of Tricholoma matsutake mushroom as an active ingredient.

In the food composition according to the present invention, the above-mentioned extract of Bombyx mori, and the bone diseases are as mentioned in the above pharmaceutical composition, respectively, unless otherwise specified.

The term " improvement " refers to any action that reduces a parameter associated with a condition for which bone disease is being treated, such as reducing the severity of symptoms, delaying the onset or deterioration of symptoms, or ameliorating symptoms.

The food may be a health functional food. The term " health functional food " means food prepared and processed in the form of tablets, capsules, powders, granules, liquids, and circles using raw materials and components having useful functions in the human body. The term "functional" as used herein means that the structure and function of the human body have a beneficial effect on health uses such as controlling nutrients or physiological actions.

The food composition according to the present invention can be prepared by a method commonly used in the art and can be prepared by adding raw materials and ingredients which are conventionally added in the art. In addition, unlike general medicines, there is an advantage that there are no side effects that may occur when a food is used as a raw material for a long time, and the portability is excellent. Therefore, the food composition of the present invention is useful for improving or improving the bone disease treatment effect It can be taken as an adjuvant.

The amount of the extract of Tricholoma matsutake extract contained as an active ingredient in the food composition according to the present invention can be suitably determined according to the intended use (prevention, improvement or therapeutic treatment). In general, in the production of food, the extract of the present invention may be contained in an amount of 0.001 to 20% by weight, 0.001 to 15% by weight or 0.001 to 10% by weight in the raw material composition. In the case of health drinks, 0.01 to 2 g, specifically 0.02 to 2 g, more specifically 0.3 to 1 g, may be added based on 100 mL. However, in the case of long-term ingestion intended for health and hygiene purposes or for the purpose of controlling health, the above amount can also be used below the above-mentioned range.

The content of the extract of Tricholoma matsutake according to the present invention added to the food composition during the preparation of the food composition can be appropriately increased or decreased as needed.

The food composition of the present invention may further include an additional ingredient in addition to the above-mentioned Samburu mallow extract for enhancing the efficacy. The additional ingredients may be, for example, minerals such as calcium, phosphorus, vitamin D, or a combination thereof, preferably calcium, phosphorus, or combinations thereof.

The food composition may be any one selected from the group consisting of pills, tablets, granules, powders, capsules, and liquid solutions.

The kind of the food is not particularly limited. 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, and includes foods in a conventional sense.

The food composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary food. The above-mentioned natural carbohydrates are sugar saccharides such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like.

When the food composition of the present invention is a beverage composition, there are no particular limitations on the liquid component other than the above-mentioned ingredients in the indicated proportions as an essential ingredient, and various flavors or natural carbohydrates, .

In addition to promoting the differentiation into osteoblasts and promoting bone formation through mineral accumulation of osteoblasts, the extract of Sanko Mackerel, which is an active ingredient of the composition according to the present invention, inhibits osteoclast differentiation of osteoclast precursors It is possible to inhibit the growth of osteoclasts, which are mononuclear cells, into osteoclasts, which are multi-nucleated cells. Accordingly, the composition according to the present invention provides fundamental treatment, prevention and improvement effects on bone diseases by promoting bone formation and inhibiting bone uptake.

FIG. 1 is a graph showing the results of an Alizarin Red assay in which the degree of accumulation of inorganic calcium in MC3T3-E1 cells was analyzed by extract of Tricholoma matsutake mushroom.
FIG. 2 is a graph showing the results of a TRAP (tartrate resistance aid phosphatase) assay in which osteoclast differentiation and inhibition of osteoclast differentiation by the extract of Tricholoma matsutake mushroom were analyzed.
Fig. 3 shows H & E staining results of the Suzuki bone and the compact bone of the mouse model. FIG. 3A is a photograph of H & E staining, FIG. 3B is a graph comparing bone mineral density of travecular bone, and FIG. 3C is a graph comparing bone thickness of a cortical bone.
Fig. 4 shows the result of masson trichrome staining of the soju bone and compact bone of a mouse model. FIG. 4A is a photograph of Masson Trichrome staining, FIG. 4B is a graph comparing bone mineral density of a travecular bone, and FIG. 4C is a graph comparing bone thickness of a cortical bone.
5 shows the MS, MS / MS and NMR data of the compound edgeworoside A isolated from the extract of Tricholoma matsutake.
6 is a graph showing the results of the Alizarin Red assay of the compound edgeworoside A isolated from the extract of Tricholoma matsutake.
FIG. 7 is a graph showing the results of TRAP assay of the compound edgeworoside A isolated from the extract of Tricholoma matsutake mori.
FIG. 8 is a graph showing the results of the toxicity and cell proliferation test of the edgeworoside A compound, MTS assay, isolated from the extract of Tricholoma matsutake mushroom.

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

Example 1 Preparation of Extract of Samjia Mulberry

1-1. Preparation of sample

Stem areas of the mulberry leaves were collected, washed with water, and then dried in a hot air drier at 40 ° C for 3 days. The stem dried material was pulverized in a pulverizer and used as an extraction sample in the subsequent extraction process.

1-2. Extraction process

30 L of methanol (methyl alcohol 99.9%) was added to 3 kg of the pulverized stem of Mulberry Mulberry, and the mixture was subjected to sonication for 15 minutes at 45 캜 for 2 hours and then repeated 10 times a day for 3 days. A liquid component was obtained. The obtained liquid phase component was concentrated under reduced pressure using a rotary evaporator (N-1000SWD; EYELA). The above extraction and concentration process was repeated twice. Finally, the dried form of the extract was extracted into a 2 ml tube and stored at 4 캜.

Example 2. Confirmation of osteoblast differentiation promoting ability of extract of Samyad mallow extract

Alkaline phosphatase (ALP) enzyme activity measurement method was used to confirm the osteoblast differentiation ability of the extract of Mulberry Mulberry.

Specifically, MC3T3-E1 cells, which are mouse cell lines, were dispensed into each well of a 384 well black clear plate at a concentration of 250 cells / 40ul, followed by overnight incubation at 37 ° C in a 5% CO ₂ incubator. After the cell culture medium was removed, the differentiation medium supplemented with 25 μg / mL of Stem Extract, 100 ng / mL of BMP2 (reference substance) and DMSO (negative control) was added to each well. And cultured in a 5% CO ₂ incubator for 14 days. Bombyx mori extract, BMP2, and DMSO were renewed every 3-4 days during the 14 day incubation period. On the last day of culture, the medium was removed from each well and the cells were washed with 50 μl of 10 mM Tris-HCl, then 20 μl of ALP was added and incubated at 25 ° C. for 1 hour. The cells were then lysed by stirring for 1 minute and then centrifuged at 1000 rpm for 1 minute. After that, 12 μl of p-nitrosophenyl phosphate (p-NPP) as a substrate of ALP enzyme was added at a concentration of 5 mM, reacted at 25 ° C. for 2 hours, and finally absorbance was measured at 405 nm using a Flexstation .

As a result, the ALP relative activity value of BMP2 (reference substance) was 100, while the ALP relative activity value of the extract of Tricholoma matsutake was 147.69. Accordingly, it was confirmed that the extract of Tricholoma matsutake mushroom of the present invention has a significant effect of promoting differentiation and activity into osteoblasts.

Example 3. Calibration of Alizarin Red assay of the extract of Tricholoma matsuri

The Alizarin Red assay was performed to examine the osteoblast activity of the extracts of mulberry mulberry. MC3T3-E1 cells were treated with Mulberry Mulberry extract for 21 days and then the amount of inorganic calcium in the bone was measured .

Specifically, MC3T3-E1 cell line (1 × 10 ⁴ cells / well) was dispensed into a 96-well plate, and osteoblast growth medium (α-MEM (Welgene, Cat. No. LM008-01), 10% FBS (Gibco 19000) , 1% penicillin-streptomycin) was added and then cultured. On the day after the incubation, the osteoblast differentiation medium (α-MEM (Welgene, cat. No. LM008-01), 1 μM dexamethasone, 50 μM ascorbic acid, 10 mM glycerophosphate) After 22 days of incubation, the cells were fixed and washed with 70% ethanol, and then ARS solution (Alizarin Red S powder (Sigma A5533), pH 4.2) was added and reacted at room temperature for 15 minutes and washed. Finally, 100 μl of cetylpyridinium chloride (100 mM) was added and reacted at 37 ° C. for 30 minutes. Absorbance was measured at 540 nm using a Flexstaion.

As a result, as shown in FIG. 1, it was observed that the calcium mineral accumulation capacity of osteoblast was improved in proportion to the concentration of the extract in the group treated with the extract of Tricholoma matsutake.

Example 4. Measurement of osteoclast differentiation and inhibition of formation formation (TRAP assay)

Tartrate-resistant acid phosphatase (TRAP) is activated when osteoclasts are differentiated in the process of bone resorption. Thus, Raw264.7 cells were treated with RANKL (Receptor activator of NF-kb Ligand) to induce differentiation into osteoclasts, and the level of TRAP produced was measured to determine osteoclast differentiation inhibitory ability of the extract.

Specifically, Raw264.7 cells were first treated with RANKL (100 ng / ml) and extracts of Mori matsutake at different concentrations for 5 days. To measure TRAP secreted in cell culture, the supernatant of the cell culture was collected, and the substrate was added to the collected supernatant. The absorbance was then measured at 540 nm using a Flexstation.

As a result, as shown in FIG. 2, about 50% of the activity of TRAP, which is an indicator of osteoclast differentiation and activity, was inhibited in the group treated with 6.25 ug / ml of the extract of Tricholoma matsutake mushroom, In the group treated with extracts, the activity of TRAP was inhibited to about 95%. Thus, it was confirmed that the differentiation and formation of osteoclast were inhibited in proportion to the treatment concentration of the extract.

Example 5: Evaluation of efficacy of extract of Sambil mulberry extract on ovariectomized animal model

Ovariectomy (OVX) was performed after 6 weeks of females ddY mice were purified for 1 week in order to confirm the inhibition and inhibition of osteoporosis in the animal model.

Specifically, animal experiments were conducted to remove the risk of animal pain and infection through safe respiratory anesthesia through isoflurane and analgesics and antibiotics. Bilateral ovariectomy blocked the normal hormone secretion of estrogen secreted from the ovaries of female ddY mice, resulting in osteoporosis with a decrease in bone density, a decrease in soju bone density, and a decrease in bone thickness.

For this experiment, as shown in Table 1 below, 5 mice were treated with Sham (non-treated group), Non_Veh (group treated only with suture after abdominal incision but without vehicle), Non_Veh OVX_Veh (vehicle-treated group after ovariectomy), OVX_GSTEP (vehicle-treated group after ovariectomy), and ovariectomized group Respectively. From the day after the operation, the extracts of Ornithoptera arees were orally administered at a dose of 10 ^-3 / kg / 10 ml once daily for 28 days.

Experimental group Treatment Number of animals Sham No treatment (no treatment) 5 grains Non_Veh Vehicle administration after abdominal incision suture 5 grains Non_GSTEP After the abdominal incision suture, Samyang mudberry extract was administered 5 grains OVX_Veh Vehicle administration after ovariectomy 5 grains OVX_GSTEP Extract of Mamdogi mudberry after ovariectomy 5 grains

After 28 days, the femur and tibia of the mouse were microcirculated by a microtome, and the resulting flakes were subjected to H & E staining and Masson trichrome staining, respectively.

As a result, as shown in Figs. 3 and 4, both the bone mineral density of the travecular bone and the bone thickness of the cortical bone decreased significantly after ovariectomy of the female mouse (see OVX_Veh group) . On the contrary, it was observed that the bone mineral density of the travecular bone was significantly increased and the cortical bone was significantly thickened when the extract of Tricholoma matsutake was administered (see OVX_GSTEP group).

Example 6. Identification of Compounds Contained in the Extract of Mulberry Mulberry

From PDMS, MS and ¹ H-NMR, it was confirmed that Edgeworthoside A, which is a compound represented by the following Formula 1, was contained in the extract from the Samburu mudberry extract prepared in Example 1:

[Chemical Formula 1]

.

.

The upper part of FIG. 5A represents ¹ H-NMR of a commercially available edgeworoside A, and the lower part of FIG. 5 (a) shows ¹ H of edgeworoside A contained in the extract of Tricholoma matsutake Lt; / RTI >

FIG. 5B shows the MS analysis result, and FIG. 5C shows the MS / MS analysis result. The top of Figures 5b and 5c represent commercially available edgeworoside A, and the bottom of Figures 5b and 5c represents edgeworoside A contained in the extract of Tricholoma matsutake .

Example 7: Efficacy of edgeworoside A compound

7.1. Calibration of Edgeworoside A Compound (Alizarin Red assay)

The calcium-accumulating ability of edgeworoside A, which was isolated from the extract of Tricholoma matsutake mushroom in Example 6, was measured in the same manner as in Example 3.

As a result, as shown in FIG. 6, it was confirmed that the accumulation capacity of calcium minerals in MC3T3-E1 cells was improved in all the experimental groups treated with the compound compared to the control group.

7.2. Determination of osteoclast differentiation and inhibition of edgeworoside A (TRAP assay)

TRAP assay was performed to measure the osteoclast differentiation and formation inhibition ability of edgeworoside A, a compound isolated from the extracts of Matsutake mushroom extract. The analysis was carried out in the same manner as in Example 4, except that Edgeworoside A, a compound isolated therefrom, was used instead of the extract.

As a result, as shown in Fig. 7, the activity of TRAP was significantly inhibited in the group treated with the compound, and in particular, in the group treated with 20 ug / ml of the compound, the activity of TRAP was inhibited by about 56% .

7.3. Toxicity test of compound edgeworoside A (MTS assay)

MTS assay was performed for the toxicity and cell proliferation of edgeworoside A, a compound isolated from the extracts of Mt.

As a result, as shown in Fig. 8, the cell survival rate was rather higher at all concentrations of the compound than the control group, and thus it was confirmed that the compound was not cytotoxic.

Claims (10)

A pharmaceutical composition for treating or preventing a bone disease, comprising an extract of Tricholoma matsutake mushroom as an active ingredient,
The bone disease is selected from the group consisting of osteoporosis, osteogenesis, osteopenia, osteoarthritis, rheumatoid arthritis, metastatic bone cancer, periodontal disease, fracture and Paget's disease,
Wherein the composition promotes osteoblast differentiation and activity and inhibits osteoclast differentiation and formation. The method according to claim 1,
Wherein the extract is a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

. The method according to claim 1,
Characterized in that the extract is obtained from the stem of the mulberry tree. A pharmaceutical composition for treating or preventing bone diseases, which comprises a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient,
The bone disease is selected from the group consisting of osteoporosis, osteogenesis, osteopenia, osteoarthritis, rheumatoid arthritis, metastatic bone cancer, periodontal disease, fracture and Paget's disease,
Wherein said composition promotes osteoblast differentiation and activity and inhibits osteoclast differentiation and formation.
[Chemical Formula 1]

. A food composition for improving or preventing osteoporosis, comprising an extract of Tricholoma matsutake mushroom as an active ingredient,
The bone disease is selected from the group consisting of osteoporosis, osteogenesis, osteopenia, osteoarthritis, rheumatoid arthritis, metastatic bone cancer, periodontal disease, fracture and Paget's disease,
Wherein the composition promotes osteoblast differentiation and activity and inhibits osteoclast differentiation and formation. 6. The method of claim 5,
Wherein the extract is a food composition comprising a compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

. 6. The method of claim 5,
Characterized in that the extract is obtained from the stem of the mulberry tree. 1. A food composition for improving or preventing bone diseases, which comprises a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient,
The bone disease is selected from the group consisting of osteoporosis, osteogenesis, osteopenia, osteoarthritis, rheumatoid arthritis, metastatic bone cancer, periodontal disease, fracture and Paget's disease,
Wherein the composition promotes osteoblast differentiation and activity and inhibits osteoclast differentiation and formation.
[Chemical Formula 1]

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