KR20190046017A - Pharmaceutical composition for use in preventing or treating osteoporosis containing camphene as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition containing camphene or a pharmaceutically acceptable derivative thereof as an active component. The pharmaceutical composition according to the present invention has an effect of promoting the differentiation of osteoblasts by increasing an expression ratio of Dlx5, Runx2, ALP, and OC, which are osteoblast differentiation marker genes, thereby being able to be widely used as a medicine or health food for preventing or treating osteoporosis. Furthermore, the pharmaceutical composition increases activity of osteoblasts, rather than targeting osteoclasts, thereby being able to fundamentally treat osteoporosis through the promotion of osteogenesis and being able to exhibit an effect of preventing secondary fracture.

Description

[0001] The present invention relates to a pharmaceutical composition for preventing or treating osteoporosis comprising Kampen as an active ingredient,

The present invention relates to a pharmaceutical composition for preventing or treating osteoporosis containing Kampen or a pharmaceutically acceptable derivative thereof as an active ingredient.

BACKGROUND ART [0002] As various industrial accidents and traffic accidents have increased and an aging society has recently been developed, musculoskeletal diseases caused by osteoporosis and fractures resulting from such diseases have been increasing. In order to prevent and treat bone diseases and the like, there have been developed various types of bone grafting agents for inducing bone regeneration ability, new types of related diseases for improving osteoclast activity and osteoclast activity. In addition, various studies using stem cells and osteoblasts have been attempted to actively induce new bone that can fill a bone tumor or bone defect.

The bone remodeling process is a process in which old bone is periodically converted into a new bone, and this process proceeds through steps such as proliferation, differentiation, and induction of calcification of the extracellular matrix. In general, the bone formation and absorption process of adult is balanced by the interaction of osteoblasts derived from mesenchymal stem cells with osteoblast-derived osteoblasts, maintaining bone health, lowering osteoblast activity and reducing bone formation Or osteoclast activity increases osteoclast activity and increases osteoclast activity, there is no significant change in the chemical composition of the osteoblast and osteoclast. However, osteopenia such as osteoporosis is caused by decreased bone mass. .

Osteoporosis is a complex disease that is also influenced by genetic factors, environmental factors such as diet and lifestyle, and especially in women, bone turnover is dramatically accelerated by hormonal changes in menopause.

Currently available osteoporosis drugs are vitamin D, female hormones, bisphosphonates, selective estrogen receptor modulators, and calcitonin preparations. Most of them are based on the way of preventing loss of bones by decreasing the activity of osteoclasts and regulating bone resorption . However, since osteoporosis treatments through inhibition of osteoclast activity are not fundamentally therapeutic methods for treating osteoporosis, there is a limit to cure and it is essential to increase osteoblast activity which can increase bone mass and bone strength.

In addition, 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 essential. Therefore, there is a need to develop new substances with superior drug efficacy without side effects when the drug is administered for a long period of time.

Recently, alternative therapies using active ingredients derived from natural products such as herbal medicines and foods that can promote bone formation with minimal side effects and minimizing bone loss have been actively conducted.

On the other hand, Camphene is a colorless camphor-like odoriferous crystal that does not dissolve in water but is soluble in organic solvents such as ethanol and ether. It is a substance mainly used for synthesis of camphor and raw materials of fragrance. Right-turn type D is contained in essential oil of leaves and stem of cedar, cypress, ginger root, stem. Left type L is contained in essential oil of pine tree leaf. It is known to exist as a component of essential oils.

Recently, skin regeneration effect, antioxidative effect, antiinflammatory effect and whitening effect of Kampen have been studied, and the effect of insecticidal insecticidal action of Sucutica causing massive mortality of fish has been studied (Patent Document 1).

However, no study has yet been reported on the effect of such Kampen on osteoblast proliferation and differentiation.

Korean Patent Registration No. 10-1298097

The inventors of the present invention have intensively tried to develop a new pharmaceutical composition which increases osteoblast activity to increase osteoporosis and increase bone mineral density and bone strength to treat osteoporosis and has less adverse effects even when administered for a long period of time. To promote the proliferation and differentiation of osteoblasts. The present invention has been accomplished on the basis of these findings.

Accordingly, an object of the present invention is to provide a pharmaceutical composition and a health functional food having excellent osteoporosis prevention and treatment efficacy and having 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 a compound represented by the following formula (1) or a pharmaceutically acceptable derivative thereof as an active ingredient.

The compound represented by Formula 1 may be referred to as camphene.

The pharmaceutical composition may be characterized in promoting osteoblast differentiation.

The concentration of the compound represented by Formula 1 or a pharmaceutically acceptable derivative thereof may be 0.01 to 20 μM.

The compound represented by Formula 1 or a pharmaceutically acceptable derivative thereof may be used in combination with any one or more of osteolysis (Runx-related transcription factor 2), Dlx5 (distal-less homeobox 5), osteocalcin (OC), and alkaline phosphatase The expression level of the cell differentiation marker gene is increased.

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

The present invention also provides a health functional food for preventing or ameliorating osteoporosis, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable derivative thereof as an active ingredient.

[Chemical Formula 1]

The health functional food may be characterized by promoting osteoblast differentiation.

The concentration of the compound represented by Formula 1 or a pharmaceutically acceptable derivative thereof may be 0.01 to 20 μM.

The pharmaceutical composition according to the present invention promotes the differentiation of osteoblast by increasing the expression level of Dlx5, Runx2, ALP, and OC, which are osteoblast differentiation marker genes, and is widely used as a medicine or a health food for prevention or treatment of osteoporosis .

In addition, rather than targeting osteoclasts, osteoblast activity can be increased to promote osteogenesis, thereby osteoporosis can be fundamentally treated, and secondary osteoporosis can be effectively prevented.

Fig. 1 is a graph showing the results of MTT analysis according to the concentration of camphene (*** P < 0.001).
FIG. 2 shows the results of Experimental Example 2 showing mRNA expression of the osteoblast differentiation gene according to the concentration of camphene.
FIG. 3 shows the result of Experimental Example 2 showing mRNA expression of the osteoblast differentiation gene according to the treatment time of camphene.
FIG. 4 shows the results of Experimental Example 2 showing mRNA expression of the osteoblast differentiation gene according to the synergistic effect of camphene.

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.

The present invention relates to a pharmaceutical composition and a health functional food for preventing or treating osteoporosis.

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 present invention provides a pharmaceutical composition for preventing or treating osteoporosis, which comprises a compound represented by the following formula (1) or a pharmaceutically acceptable derivative thereof as an active ingredient.

[Chemical Formula 1]

The compound represented by Formula 1 may be referred to as camphene.

The compound represented by the formula (1) can be used in the form of a pharmaceutically acceptable salt. As the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid and the like, aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, Derived from organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid and the like. Examples of such pharmaceutically innocuous salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, But are not limited to, but are not limited to, but are not limited to, but are not limited to, but are not limited to, halides, halides, halides, halides, halides, halides, But are not limited to, lactose, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chloro Such as benzenesulfonate, benzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate,? -Hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene- 1-sulfonate, naphthalene-2-sulfonate, mandelate and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the derivative of Chemical Formula 1 in an organic solvent such as methanol, ethanol, acetone, dichloromethane, acetonitrile and the like, Followed by filtration and drying, or by distillation of the solvent and excess acid under reduced pressure, followed by drying and crystallization in an organic solvent.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. In addition, the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

The compound represented by the above formula (1), Kamppen can be extracted from a natural product or synthesized by a known method, and its production method and origin are not limited.

Kampen is an essential oil component of various plants such as pine, pine, mountain pepper, shiitake, and camphor tree. It is known to have various effects such as sedation, diuretic, giant wind, hypertension, edema, insomnia and fat loss.

The pharmaceutical composition may be characterized in promoting osteoblast differentiation.

Compounds that promote osteoclast differentiation can be used not only for the prevention or treatment of diseases and symptoms such as osteoporosis and fracture but also for the treatment of periodontal disease for the purpose of bone strengthening (Zhang et al., Shanghai Kou Qiang Yi It has also been reported that it may be useful for the treatment of bone growth disorders resulting from burns, as reported by Xue 7 (2), pp 99-103, 1998; Cahill et al, Biol Blood Marrow Transplant 10 (10), pp709-717, (Klein et al., Osteoporos Int. 16 (6), pp. 631-635, 2005).

The pharmaceutical composition can be used without limitation in diseases causing bone formation due to deficiency of osteoblasts and the like. More specifically, the pharmaceutical composition can be used for the treatment of bone destruction and inflammatory diseases caused by interleukin-1 (IL-1) And can be used to treat osteoporosis, bone metastatic lesion, primary bone tumor, rheumatic or degenerative arthritis, periodontal disease, inflammatory alveolar bone disease, inflammatory bone resorption disease And Paget's &lt; Desc / Clms Page number 2 &gt; disease, and the like.

The concentration of the compound represented by Formula 1 or a pharmaceutically acceptable derivative thereof may be 0.01 to 20 μM.

If the concentration is less than 0.01 μM, the pharmacological effect may be insignificant. If the concentration is more than 20 μM, cytotoxicity may occur.

The compound represented by Formula 1 or a pharmaceutically acceptable derivative thereof may be used in combination with any one or more of osteolysis (Runx-related transcription factor 2), Dlx5 (distal-less homeobox 5), osteocalcin (OC), and alkaline phosphatase The expression level of the cell differentiation marker gene is increased.

Osteoblast differentiation can be induced by a combination of hormones, cytokines such as bone morphogenetic proteins (BMPs), runt-related transcription factor 2 (Runx2), inhibitor of DNA binding 1, Dlx5 (distal-less homeobox 5) (osteocalcin), and alkaline phosphatase (ALP). Among them, Id1, Dlx5, and Runx2 are essential genes that are expressed at the early stage of osteoblast differentiation. Expression of these genes means that osteoblast differentiation is accelerated.

Runx-related transcription factor 2 (Runx2) is known to be an important regulator of osteoblast differentiation, and it is known that ALP (alkaline phosphatase), OC (osteocalcin) and bone sialoprotein ) To control osteoblast differentiation.

Id1 (inhibitor of DNA binding 1) is a transcription factor that affects the balance between cell growth and differentiation in osteoblasts and is known to play an important role in the early stages of osteoblast-specific differentiation. In addition, differentiation is promoted by the regulation of Id1 (inhibitor of DNA binding 1) gene expression when transformed into osteoblast differentiation from myoblast and mesenchymal stem cells.

Dlx5 (distal-less homeobox 5) is also induced by bone and directly binds to the promoter of Runx2 (runt-related transcription factor 2) to regulate transcription, and gene expression of OC (osteocalcin), the late marker gene of osteoblast differentiation To control osteoblast differentiation.

The pharmaceutical composition 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 pharmaceutical 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.

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.

The present invention also provides a health functional food for preventing or ameliorating osteoporosis, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable derivative thereof as an active ingredient.

[Chemical Formula 1]

The compound represented by Formula 1 may be referred to as camphene.

The health functional food may be characterized by promoting osteoblast differentiation.

The concentration of the compound represented by Formula 1 or a pharmaceutically acceptable derivative thereof may be 0.01 to 20 μM.

If the concentration is less than 0.01 μM, the pharmacological effect may be insignificant. If the concentration is more than 20 μM, cytotoxicity may occur.

The health functional food may be manufactured in the form of, but not limited to, various drinks, gum, tea, confectionery, vitamin complex, health supplement, and the like.

In addition, the health functional food includes a case where it is added to health food for the purpose of improving osteoporosis, and there is no particular limitation on the kind of 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 amount of the health functional food to be consumed depends on the condition and the weight of the recipient, the degree of symptoms, the type of food, the period of consumption, and may be appropriately selected.

The differentiation of osteoblasts during bone formation is controlled by the expression of genotypes and has inherent characteristics depending on the culture method. Important signaling pathways involved in osteocyte differentiation and osteogenesis are transforming growth factor-beta (TGF-beta), bone morphogenetic protein (BMP), and beta-catenin , Fibroblast growth factor (FGF), hedgehog, and notch are known.

These signaling pathways induce the expression and activation of various transcription factors related to osteogenesis such as Runx2 (Runt-related transcription factor 2) and Dlx5 (Distalless-related homeobox 5) during osteoclast differentiation, and ALP (alkaline phosphatase) , Id1 (inhibitor of DNA binding 1), and OC (osteocalcin). As a differentiation inducer in such osteoblasts, ascorbic acid (AA), beta-glycerophosphate (beta -GP), dexamethasone and the like are known.

In the present invention, experiments were carried out to investigate the effect of camphene on the proliferation and differentiation of osteoblast cells using MC3T3-E1 preosteoblasts similar to osteoblasts present in bone tissue. As a result, Proliferation and differentiation of the cells.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, and thus are not limitative of the present invention, It is provided for a complete description.

< Experimental Example  1> Cytotoxicity experiment

1-1. Experimental Method

(1) Experimental materials and cell culture

The cells were cultured in MC3T3-E1 cells (ATCC, Manassas, Va., USA), which was derived from mouse, and alpha-minimal essential (Α-MEM, Sigma aldrich) and phosphate buffer saline (PBS, Gibco, Grand Island, NY, USA) were used. Fetal bovine serum (FBS) was purchased from ATLAS, except for the reagents purchased from Sigma Aldrich Korea.

MC3T3-E1 (osteoclasts) were cultured in alpha-minimal essential nutrient medium (α-MEM) supplemented with 1% penicillin-streptomycin (P / S) and 10% fetal bovine serum (FBS) After sterilization, 0.25% Trypsin EDTA (Gibco) solution was used to subculture in an environment of 37 ° C, 5% CO ₂ or less, and the culture medium was changed every 2 days for the experiment. Ascorbic acid and β-glycerophosphate (β-GP) at a concentration of 50 μg / ml were used for the control of the experiment. Were cultured and treated.

(2) MTT analysis

 (MTT) (Sigma-Aldrich Co., St. Louis, Mo., USA) was used to measure cytotoxicity ) Was used to measure the survival rate of living cells.

MC3T3-E1 cells were subcultured in each well of a 48-well plate to allow cells to enter, and then cultured for 24 hours so that the cells could be adhered. Then, campin was added to each concentration (10, 20, 50, 100, 200 μM) and cultured.

 As MTT assay, cells cultured with various concentrations of Kampen were treated with 10% MTT culture solution at a concentration of 5 mg / mL and cultured in a 37 ° C incubator for 1 hour. When the cells were stained with purple dye, 400 μl of dimethyl sulfoxide (DMSO) was added to the 96-well plate, and 100 μl of the solution was dispensed. The absorbance of the solution was measured at 540 nm using an analyzer.

1-2. Experiment result

The results of MTT analysis conducted in MC3T3-E1 cells to confirm the cytotoxicity of camphene are shown in FIG.

As shown, no cytotoxicity was observed in Kampen at concentrations below 20 μM, but toxicity was seen at concentrations above 50 μM.

< Experimental Example  2> Osteoblast differentiation by camphene treatment Marker  Expression analysis

2-1. Experimental Method

(1) RNA extraction

For RNA extraction, 300μl of triazole was treated in a 6-well plate, and the cells were collected with scrapers and allowed to react for 10 minutes. Then, 60 μl of chloroform was added and reacted for another 10 minutes.

 After centrifugation (14500 rpm, 4 캜, 15 min), the RNA portion of the supernatant was transferred, treated with 150 μl of isopropyl and reacted at 4 ° C for 30 minutes. After centrifugation (14500 rpm, 4 캜, 10 min), the supernatant was removed and the pellet was washed with 70% ethanol and centrifuged again (13000 rpm, 4 캜, 10 min). After removing ethanol, it was dried for 20 minutes and dissolved in tertiary distilled water at 4 ° C for 30 minutes.

(2) cDNA synthesis

For cDNA preparation, the absorbance was measured at 260-280 nm and the amount of RNA, tertiary distilled water, and Kit (TopScript ™ RT Drymix) (Enzynomics, Daejeon, Korea) was adjusted to 25 μl. Finally, cDNA synthesis was carried out at 42 ° C for 60 minutes and at 95 ° C for 5 minutes.

(3) Real Time Polymerase Chain Reaction (RT-PCR)

1 μl of cDNA and 2 μl of each primer were mixed with 10 μl of Emerald Green mixture (Takara Bio, Otsu, Japan) and 7 μl of third distilled water to perform real-time PCR. The nucleotide sequences of the respective primers and the experimental conditions are shown in Tables 1 and 2. After that, electrophoresis was performed using 1% agarose gel, and the resulting band was measured by Gel doc (Alpha innotech corporation, San Francisco, USA).

gene
Primer Sequence Forward Reverse β-actine 5'-TTC TTT GCA GCT CCT TCG TTG CCG-3 ' 5'-TGG ATG GCT ACG TAC ATG GCT GGG-3 ' Runx2 5'-CCG CAC GAC AAC CGC ACC AT-3 ' 5'-CGC TCC GGC CCA CAA ATC TC-3 ' Dlx5 5'-CAG AAG AGT CCC AAG CAT CC-3 ' 5'-GAG CGC TTT GCC ATA AGA AG-3 ' OC 5'-GCA AAT AAG GTA GTG AAC AGA CTC C-3 ' 5'-GTT TGT AGG CGG TCT TCA AGC-3 ' ALP 5'-ATC TTT GGT CTC GCT CCC ATG-3 ' 5'-TTT CCC GTT CAC CGT CCA C-3 '

gene
Reaction conditions Cycle Number Pre-
Denaturation Denaturation Annealing Extention Post-
Extension β-actine 25 cycles

95 ° C 5 min



95 ° C 30 sec

66 ° C 30 sec

72 ° C 30 sec



72 ° C 5 min

Runx2
30 cycles
62 ° C 30 sec Dlx5 63 ° C 30 sec OC 60 ° C 30 sec ALP 35 cycles

2-2. Experiment result

(1) mRNA expression of osteoblast differentiation gene according to the concentration of camphene

Experiments showing the mRNA expression level of the osteoblast differentiation gene were carried out using Runx2, Dlx5 and ALP at a concentration of 10, 20 μM according to the cytotoxicity test results. The results are shown in FIG.

As shown, an increase in the expression level was observed at a concentration of 10, 20 μM of Kampen compared to the control group. In addition, it was confirmed that the expression level of all mRNA was increased at 10 μl more than 20 μl.

(2) Expression of mRNA of osteoblast differentiation gene according to processing time of camphene

The mRNA expression of the osteoblast differentiating gene according to the concentration of Kampen was most strongly expressed at a concentration of 10 μM. Thus, experiments showing the degree of mRNA expression by treatment with Kampen at 10 μM of Kampen concentration were performed at Runx2, Dlx5 , ALP, and OC. The results are shown in FIG.

As shown, when compared with the control group, there was no increase in expression amount in a time-dependent manner, but it was confirmed that a definite increase was observed.

(3) mRNA expression of osteoblast differentiation gene according to synergistic effect of camphene

(Ascorbic acid + beta-glycerophosphate, CP: camphene, AA + beta-GP + CP: ascorbic acid + beta-glycerophosphate + camphene) in order to examine the synergistic effect of Kampen And then experiments were conducted to show the degree of mRNA expression using Runx2, Dlx5, ALP, and OC. The results are shown in FIG.

As shown, mRNA expression of AA + β-GP and camphene was higher than that of independent AA + β-GP mRNA expression.

Meanwhile, the compound represented by Formula 1 according to the present invention can be formulated into various forms according to the purpose. Hereinafter, some formulation methods in which the compound represented by Formula 1 according to the present invention is contained as an active ingredient are exemplified, and the present invention is not limited thereto.

<Formulation example>

1-1. Manufacture of Powder

500 mg of the compound of formula (1)

Lactose 100 mg

10 mg of talc

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

1-2. Manufacture of tablets

500 mg of the compound of formula (1)

Corn starch 100 mg

Lactose 100 mg

2 mg of magnesium stearate

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

1-3. Manufacture of capsules

500 mg of the compound of formula (1)

Corn starch 100 mg

Lactose 100 mg

2 mg of magnesium stearate

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

1-4. Injection preparation

500 mg of the compound of formula (1)

Sterile sterilized water for injection

pH adjuster

(2 ml) per ampoule in accordance with the usual injection method.

1-5. Manufacture of liquid agent

100 mg of the compound of formula (1)

10 g per isomer

5 g mannitol

Purified water quantity

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, The liquid is prepared by sterilization.

Claims (8)

1. A pharmaceutical composition for preventing or treating osteoporosis, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable derivative thereof as an active ingredient.
[Chemical Formula 1]

The method according to claim 1,
A pharmaceutical composition for preventing or treating osteoporosis, wherein the pharmaceutical composition promotes osteoblast differentiation.
The method according to claim 1,
A pharmaceutical composition for preventing or treating osteoporosis, wherein the concentration of the compound represented by the formula (1) or a pharmaceutically acceptable derivative thereof is 0.01 to 20 μM.
The method according to claim 1,
The compound represented by Formula 1 or a pharmaceutically acceptable derivative thereof may be used in combination with any one or more of osteolysis (Runx-related transcription factor 2), Dlx5 (distal-less homeobox 5), osteocalcin (OC), and alkaline phosphatase Wherein the expression level of the cell differentiation marker gene is increased.
The method according to claim 1,
Wherein said pharmaceutical 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, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable derivative thereof as an active ingredient.
[Chemical Formula 1]

The method according to claim 6,
Wherein said health functional food promotes osteoblast differentiation.
The method according to claim 6,
Wherein the concentration of the compound represented by Formula 1 or a pharmaceutically acceptable derivative thereof is 0.01 to 20 μM.

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