WO2017209410A1 - Novel compound promotive of osteoblast differentiation and inhibitory of adipose cell differentiation, preparation method therefor, and application thereof - Google Patents

Abstract

The present invention relates to a compound having the activity of promoting the differentiation of osteoblasts and inhibiting the differentiation of adipose cells, and a preparation method therefor. A novel compound of the present invention increases the expression of the gene ALP involved in the differentiation of osteoblasts, regulates the expression of the genes PPARγ, aP2, and CD36 involved in the differentiation of adipose cells, increases a bone mass density (BMD)in an ovariectomized osteoporous animal model, and decreases a level of adipose cells in the bone marrow. Thus, the compound can be used as an effective ingredient of medications or health functional foods for bone metabolic diseases or obesity.

Description

Novel compounds that promote osteoblast differentiation and inhibit adipocyte differentiation, methods of preparation and application thereof

The present invention relates to a novel compound that promotes osteoblast differentiation and inhibits adipocyte differentiation, and a method for producing the same. The novel compound of the present invention can be used in the manufacture of a dietary supplement or a medicine related to bone metabolism disease or obesity.

Bones not only structurally support muscles or organs, but also support the soft tissues and weight of the human body, surround internal organs, protect internal organs from external shocks, and store substances such as calcium and other essential minerals in the body, such as phosphorus or magnesium. One of the important parts of the human body.

Bone is composed of osteoblasts (osteoblast), osteocytes (osteocyte), osteoclasts (osteoclast). Among them, osteoblasts are derived from mesenchymal stem cells capable of differentiating into chondrocytes, myocytes, and adipocytes, and form bone tissues through proliferation, bone matrix formation, and calcification. It plays a role. The osteoclasts also play a role in absorbing bone.

The bones of grown adults continue to regenerate and regenerate the bone absorption and production of osteoclasts, which remove old bones and replace osteoblasts with new ones. For example, osteoblasts regulate homeostasis of bone metabolism in the body by regulating the differentiation of osteoclasts responsible for bone resorption through the secretion of substances such as receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG). Keep it.

When the homeostasis of the bone metabolism is broken by a specific cause, a bone metabolic disease such as osteoporosis, bone formation disorder or fracture occurs.

Osteoporosis, a typical bone metabolic disease, is characterized by a decrease in bone mass and bone quality, with bone mineral density of 2.5 or less, or T-score (standard deviation from the average adult's average bone mass) of -2.5 or less. Say the status. Osteoporosis occurs frequently in postmenopausal women, and with age, the bone matrix decreases and fat cells form in the voids, and the formed fat cells decrease the function and differentiation of osteoblasts, which form bone, and release inflammatory cytokines. It is known to promote the function and differentiation of osteoclasts, which are responsible for the absorption of bone by secretion. If the bone density is excessively reduced, a small impact will easily cause fractures. Osteoporosis is not a symptom itself but rather various fractures caused by bone weakness, especially femoral fractures or vertebral fractures, which limit long-term activity and lead to a healthy life, resulting in 15% of elderly deaths. Known.

Osteodystrophy is also called osteotrophy and is a disease of bone caused by chronic kidney failure. It is caused by congenital abnormal kidney function and dies when dialysis is not performed when the kidney is weak. This bone disease is called renal osteodystrophy. Bone diseases related to osteotrophy include osteomalacia and osteotease fibrosa.

Calcium adjuvant is recommended for the treatment or prevention of the bone metabolism disease, vitamin D, or hormones such as estrogen or calcitonin are recommended for postmenopausal women. In addition, a bisphosphonate family such as Fosamax (component name: alendronate) and Actonel (component name: risedronate) is mainly used for bone resorption inhibitors that inhibit osteoclasts and induce death.

However, calcium adjuvant inhibits secretion of parathyroid hormone and prevents bone loss due to bone resorption, but individual differences in bone mass maintenance are known to be severe. Hormone increases bone density, but side effects such as breast cancer, myocardial infarction and venous thrombosis (Nelson, HD et al., JAMA, 288: 872-881, 2002; Lemay, A., J. Obstet. Bynaecol. Can., 24: 711-7152-3). In the case of bisphosphonate preparations, cases of necrosis of the jawbone, severe atrial fibrillation, incapacitation of bones or joints or pain of the musculoskeletal are reported (Coleman RE., Br J Cancer, 98: 1736-1740 (2008).

In addition, the conventional agents for the treatment or prevention of bone metabolic diseases have a pharmacological action to prevent bone loss anymore, but to date, there was no effect to restore the reduced bone mass to its original state.

Accordingly, the present inventors have confirmed that the newly synthesized new compounds can treat and prevent bone metabolic diseases or obesity through the regulation of differentiation of osteoblasts and adipocytes and completed the invention.

An object of the present invention is to provide a composition for the prevention or treatment of bone metabolic diseases comprising the new compound as an active ingredient.

Another object of the present invention to provide a health functional food for improving or preventing bone metabolism disease using the new compound as an active ingredient.

Another object of the present invention is to provide a novel compound having the activity of promoting osteoblast differentiation and inhibiting adipocyte differentiation.

Another object of the present invention is to provide a method for preparing a novel compound that has the activity of promoting osteoblast differentiation and inhibiting adipocyte differentiation.

The composition for preventing or treating bone metabolism disease of the present invention is a compound of the formula (1), an isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the health functional food for improving or preventing bone metabolism disease of the present invention is a compound of Formula 1, an isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the compound having the activity of promoting osteoblast differentiation and inhibiting adipocyte differentiation of the present invention is a compound of Formula 1 below.

[Formula 1]

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and R ₆ may be the same as or different from each other, and each independently hydrogen, hydroxy, alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, Any one selected from the group consisting of halogen and trifluoromethyl,

R ₇ , R ₈ , R _9, and R ₁₀ may be the same as or different from each other, and each independently hydrogen or a phenyl group, and when R ₇ and R ₉ are hydrogen, R ₈ and R ₁₀ are phenyl, and R _{If 7} and R ₉ are phenyl then R ₈ and R ₁₀ are hydrogen and the phenyl is unsubstituted or substituted with any substituent selected from the group consisting of hydroxy, halogen and trifluoromethyl.

Preferably, R ₁ , R ₃ , R ₄ and R ₆ may be the same or different from each other, and each independently one selected from the group consisting of alkyl having 1 to 6 carbon atoms and alkoxy having 1 to 6 carbon atoms. ,

R ₂ and R ₅ may be the same as or different from each other, and each independently one selected from the group consisting of hydrogen, hydroxy, halogen, and trifluoromethyl.

More preferably, the compound of Formula 1 is a compound of Formula 7, Formula 8, or Formula 9.

[Formula 7]

[Formula 8]

[Formula 9]

Producing a compound of Formula 6 by promoting osteoblast differentiation of Formula 7 and having an activity of inhibiting adipocyte differentiation, reacting a compound of Formula 4 with a compound of Formula 5; And removing the MOM-protecting group from the compound represented by Chemical Formula 6;

[Formula 4]

[Formula 5]

[Formula 6]

Producing a compound of Formula 3 by promoting osteoblast differentiation of Formula 7 and having an activity of inhibiting adipocyte differentiation by reacting a compound of Formula 2 with chloromethylmethyl ether; And preparing the compound of Chemical Formula 4 by reacting the compound of Chemical Formula 3 with diisobutyl aluminum hydride.

[Formula 2]

[Formula 3]

The compound of Formula 2 may be synthesized from 4-hydroxy-3,5-dimethoxycinnamic acid.

'MOMO-' of Chemical Formulas 3 to 6 exemplarily show that methoxymethyl, which is one of the hydroxyl-protecting groups, is ester-bonded to the hydroxyl group, and the hydroxyl protecting group is a desired chemical in the molecule. After the reaction is completed, it relates to a functional group that is easy to remove and suitable for protecting the hydroxyl group against chemical reactions. Typically such functional groups are the aforementioned unsubstituted or substituted aryl, aralkyl or acyl groups, additionally alkyl groups. The nature and size of the hydroxyl protecting groups is not critical for their removal after the desired chemical reaction or reaction sequence: Preferably they can be functional groups having 1-20 carbon atoms, in particular 1-10. The hydroxy-protecting groups are for example benzyl, methoxymethyl, 4-methoxybenzyl, p-nitro-benzoyl, p-toluenesulfonyl, tert-butyl and acetyl, with benzyl and methoxymethyl being particularly preferred Do.

As used herein, the term 'pharmaceutically acceptable salt' refers to a formulation of a compound that does not cause serious irritation to the organism to which the compound is administered and does not impair the biological activity and properties of the compound. The pharmaceutically acceptable salts include acids that form non-toxic acid addition salts containing pharmaceutically acceptable anions, such as inorganic acids, such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, and tartaric acid. Organic carboxylic acids such as formic acid, citric acid, acetic acid, trichloroacetic acid, trichloroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, salicylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfate Acid addition salts formed by sulfonic acids and the like such as phonic acid and the like. For example, pharmaceutically acceptable carboxylic acid salts include metal salts or alkaline earth metal salts formed by lithium, sodium, potassium, calcium, magnesium, amino acid salts such as lysine, arginine, guanidine, dicyclohexylamine, N Organic salts such as -methyl-D-glucamine, tris (hydroxymethyl) methylamine, diethanolamine, choline and triethylamine and the like.

As used herein, the term 'as an active ingredient' means containing an amount sufficient to achieve the efficacy or activity of the compound of Formula 1.

The bone metabolic disease includes osteoporosis, bone dysplasia or fractures.

The 'osteoporosis' includes all types of clinical classification according to bone mineral density (BMD) measurement, that is, osteopenia, osteoporosis, and severe osteoporosis. In addition, primary osteoporosis includes type 1 postmenopausal osteoporosis, type 2 senile osteoporosis, and secondary osteoporosis.

The `` osteodystrophy '' includes osteomalacia, osteote fibrosa, and the like.

The term 'fracture' refers to a state in which continuity of bone or cartilage is completely or incompletely lost or linear deformation occurs. The fractures are repeatedly loaded with pathological fractures caused by anatomical location, degree of fracture, direction of fracture surface, presence of open window, number of fractures, stability, presence of fracture fragments, osteoporosis and tumor osteomyelitis, which are special fractures. It can be classified as fatigue fracture caused by the addition.

The composition for preventing or treating bone metabolism disease of the present invention promotes osteoblast differentiation and has an activity of inhibiting adipocyte differentiation. Therefore, obesity and bone metabolic diseases can be prevented or treated simultaneously.

The composition for preventing or treating bone metabolism disease of the present invention may include a drug, vitamin, natural product, or extract thereof having the prophylactic or therapeutic activity of bone metabolic disease together with the compound of Formula 1. For example, with the compound of Formula 1, calcium adjuvant, vitamin D, hormonal agents such as estrogen or calcitonin, fosamax (component name: alendronate), and bisphosphonate-based bone such as actonel (component name: risedronate) Absorption inhibitors may be included in combination of one or more.

The composition for preventing or treating bone metabolic disease of the present invention may be prepared by including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients. Pharmaceutically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, if necessary, including antioxidants, buffers, Other conventional additives such as bacteriostatic agents can be added. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA.

The composition for preventing or treating bone metabolic disease of the present invention can be administered orally or parenterally (eg, applied intravenously, intraperitoneally, subcutaneously or topically) according to the purpose, and the dosage is weight, age, sex of the patient. The range varies depending on health, diet, time of administration, method of administration, rate of excretion and severity of disease. In the present invention, the dosage of the compound of formula I is 0.1 mg / kg to 10 g / kg per day, preferably 1 mg / kg to 1 g / kg. Administration can be administered once a day or divided into several times depending on the purpose.

Health functional foods for improving or preventing bone metabolism disease of the present invention promotes osteoblast differentiation and has the activity of inhibiting adipocyte differentiation. Therefore, obesity and bone metabolic diseases can be simultaneously improved or prevented.

Health functional foods for improving or preventing bone metabolism disease of the present invention may include vitamins, natural products, or extracts thereof having the improvement or prophylactic activity of the bone metabolism disease with the compound of Formula 1. For example, one or more calcium adjuvant, vitamin D, etc. may be included in combination with the compound of Formula 1.

Health functional foods for improving or preventing bone metabolism disease of the present invention may include food supplements additives in addition to foods acceptable in addition to the active ingredient described above.

The health functional food of the present invention includes various foods, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages.

When the compound of Formula 1 is added to a food or drink for the purpose of improving or preventing bone metabolism disease, the amount of the compound in the food may generally be added in 0.001 to 5% by weight of the total food weight, the amount of the compound in the beverage It can be added at a rate of 0.002 to 5 g, preferably 0.03 to 1 g, based on 100 ml of the beverage.

In addition to containing the compound of Formula 1 as an active ingredient, the beverage does not have any particular limitation, and may contain various flavors or natural carbohydrates as additional components, as in general beverages. Examples of the above-mentioned natural carbohydrates are conventional monosaccharides such as disaccharides such as glucose and fructose, such as maltose, sucrose and the like, and polysaccharides such as dextrin, cyclodextrin and the like. Sugars and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of beverage.

In addition to the above, the health functional food of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the health functional foods of the present invention may contain fruit flesh for the production of natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the health functional food of the present invention.

The present invention relates to a compound having an activity for promoting osteoblast differentiation and inhibiting adipocyte differentiation, and a method for preparing the same. The novel compound of the present invention increases the expression of ALP, a gene involved in the differentiation of osteoblast, Drugs or health functions that regulate the expression of PPARγ, aP2 and CD36, genes involved in differentiation, increase BMD in ovarian osteoporosis animal models, and reduce fat cells in the bone marrow, resulting in bone metabolism or obesity It can be used as an active ingredient of food.

1 is a ¹ H NMR spectrum of a NK11 compound.

2 is a ¹³ C NMR spectrum of a NK11 compound.

Figure 3 is an ALP staining photograph for confirming the change in the production of ALP (alkaline phosphatase) according to the treatment of NK11 compound when osteoblast differentiation of C3H10T1 / 2 cell line in Experimental Example 1, the ALP production change is a graph measured by -b value .

Figure 4 is the result of oil red o staining to show the change in fat production according to the NK11 compound treatment when adipocyte differentiation of C3H10T1 / 2 cell line in Experimental Example 1, the staining result is a graph showing the oil red o absorbance.

Figure 5 is a graph showing the expression changes of the mRNA expressions PPARγ, aP2, CD36, ALP in the C3H10T1 / 2 cell line according to the NK11 compound treatment in Experimental Example 1.

6 is an ALP staining photograph for comparing the change in the production of ALP (alkaline phosphatase) according to the treatment of the NK11 compound and the compound of Formula 8 during osteoblast differentiation of C3H10T1 / 2 cell line in Experimental Example 2, the change in the ALP production amount -b value It is a graph measured by.

7 is a result of staining with oil red o to show a change in fat production amount according to the treatment of NK11 compound and the compound of formula 8 when osteoblast differentiation of C3H10T1 / 2 cell line in Experimental Example 2, and the staining result is oil red o absorbance It is a graph.

8 is a graph showing changes in the expression level of mRNA PPARγ, aP2, CD36, ALP in the C3H10T1 / 2 cell line according to the compound of Formula 8 in Experimental Example 2.

9 is ALP for treating the compounds of Formula 4, Formula 2, Formula 10, Formula 11, and Formula 9 during osteoblast differentiation of C3H10T1 / 2 cell line in Experimental Example 3 to compare the changes in ALP (alkaline phosphatase) production with NK11 compounds. It is a stained picture, and is a graph measuring the change in the ALP production amount by -b value.

FIG. 10 shows oil red o staining in order to compare fat production with NK11 compounds by treating compounds of Formula 4, Formula 2, Formula 10, Formula 11 and Formula 9 in adipocyte differentiation of C3H10T1 / 2 cell line in Experimental Example 3. It is a result, and the staining result is a graph showing the oil red o absorbance.

FIG. 11 compares the expression changes of mRNAs of PPARγ, aP2, CD36, and ALP in the C3H10T1 / 2 cell line according to the treatment of the compounds of Formulas 4, 2, 10, 11, and 9 in Experimental Example 3 with NK11 compounds. It is a graph.

Figure 12 is a graph comparing the weight of ovarian resection mice administered orally for 8 weeks with the NK11 compound in Experimental Example 4.

Figure 13 is a graph measuring the concentration of triglycerides, cholesterol, ALP in plasma of ovarian resection rats administered NK11 compound in Experimental Example 4.

Figure 14 is a graph measuring the BMD of the ovarian resected rat femur to which the NK11 compound was administered in Experimental Example 4.

15 is a graph comparing mRNA expression levels of osteoblasts in ovarian resected rat femurs to which the NK11 compound was administered in Experimental Example 4. FIG.

Hereinafter, the present invention will be described in detail by using examples and experimental examples. However, the following Examples and Experimental Examples are merely illustrative of the present invention, but the content of the present invention is not limited thereto.

Production Example  1: compound of formula 3 ( dimethyl  2,5-bis (3,5- dimethoxy Synthesis of -4- (methoxymethoxy) phenyl) tetrahydrofuran-3,4-dicarboxylate

The compound of formula (II) was prepared using a known synthesis method from 4-hydroxy-3,5-dimethoxycinnamic acid.

Put the furan compound of formula 2 (401 mg, 0.812 mmol) in a 10 mL round bottom flask and dichloromethane (CH ₂ Cl ₂ , 3 mL), diisopropylethylamine (495 μL, 2.84 mmol) under nitrogen gas. And chloromethylmethyl ether (chloromethylmethylether, 216 μL, 2.843 mmol) was added. After stirring for 12 hours at room temperature, an aqueous ammonium chloride solution (5 mL) was added to the reaction solution to quench the reaction. Anhydrous magnesium sulfate was added to the organic layer solution obtained by extracting three times with 5 mL of dichloromethane to remove water, and then filtered and concentrated. The concentrated reaction product was separated by silica gel column chromatography (hexanes: EtOAc, 2: 1) to give dimethyl 2,5-bis (3,5-dimethoxy-4- (methoxymethoxy) phenyl) tetrahydrofuran, a compound of formula 3 as a yellow liquid. -3,4-dicarboxylate (476 mg, 0.819 mmol, 99% yield) was synthesized.

IR (neat): 2983 (w), 1735 (s), 1365 (m), 1240 (s), 1045 (s), 920 (m), 732 (s) cm-1;

¹ H NMR (CDCl ₃ , 400 MHz): δ 6.68 (s, 4H), 5.38 (d, J = 2.5 Hz, 1H), 5.36 (d, J = 2.5 Hz, 1H), 5.13 (s, 4H), 3.87 (s, 12H), 3.74 (s, 6H), 3.61 (s, 6H), 3.60 (d, J = 2.4 Hz, 2H);

¹³ C NMR (CDCl ₃ , 100 MHz): δ 171.5, 153.3, 135.9, 133.9, 102.5, 97.9, 82.9, 56.8, 56.2, 55.8, 52.3;

HRMS (ESI-TOF) m / z: [M + Na] + Calcd for C 28 H 36 NaO 13 603.2054; Found 603.2059.

Production Example  2: compound of formula 4 (2,5- bis (3,5-dimethoxy-4- Synthesis of (methoxymethoxy) phenyl) tetrahydrofuran-3,4-diyl) dimethanol)

Methyl ester of MOM-protecting group (50.0 mg, 0.0861 mmol) was added to a 10 mL round bottom flask, toluene (1.9 mL) was added under nitrogen gas and diisobutylaluminum hydride (DIBAL-H) (420 μL, 0.419 mmol) was added. After stirring for 1 hour at room temperature, 3 mL of potassium sodium tartrate aqueous solution was slowly added to the prepared reaction solution to quence the remaining aluminum hydride, and then washed three times with ethyl acetate (3 mL x 3). The organic layer solution was collected, water was removed with anhydrous magnesium sulfate, filtered and concentrated. The concentrated reaction product was separated by silica gel column chromatography (CH2Cl2: MeOH, 4: 1) to give a colorless liquid alcohol 2,5-bis (3,5-dimethoxy-4- (methoxymethoxy) phenyl) tetrahydrofuran. -3,4-diyl) dimethanol (43.0 mg, 0.0827 mmol, 96% yield) was synthesized.

IR (neat): 3392 (br s), 2940 (w), 1591 (m), 1227 (m), 1114 (s), 957 (s), 838 (w) cm-1;

¹ H NMR (CDCl ₃ , 400 MHz): δ 6.61 (s, 4H), 5.12 (s, 4H), 4.78 (d, J = 8.8 Hz, 2H), 3.87 (s, 12H), 3.86-3.82 (m , 2H), 3.71-3.63 (m, 2H), 3.60 (s, 6H), 3.40 (br s, 2H), 2.35-2.26 (m, 2H);

¹³ C NMR (CDCl ₃ , 100 MHz): δ 153.6, 137.8, 134.3, 103.3, 98.2, 83.4, 63.0, 57.1, 56.7, 56.2;

HRMS (ESI-TOF) m / z: [M + Na] + Calcd for C 26 H 36 NaO 11 547.2155; Found 547.2158.

Production Example  3: compound of formula 6 (( 2E, 2'E ) -2,5- bis (3,5-dimethoxy-4- (methoxymethoxy) phenyl) tetrahydrofuran-3,4-diyl) bis (methylene) bis (3- (4- (methoxymethoxy) phenyl) acrylate))

Compound (4) (580 mg, 1.11 mmol) substituted with two alcohol groups in a 100 mL round bottom flask, transcoumarinic acid derivative (575 mg, 2.76 mmol), DMAP (4-dimethylaminopyridine, 405 mg, 3.32) mmol) and EDC ( N- (3-dimethylaminopropyl) -N -ethylcarbodiimide hydrochloride, 529 mg, 2.76 mmol) were added thereto, and the reaction temperature was reduced to 0 ° C. under nitrogen gas. Then, 61 mL of dichloromethane was added thereto. After stirring at 0 ° C. for 1 hour, the temperature was raised to room temperature, followed by further stirring for 2 hours. Ammonium chloride solution (60 mL) was added to the reaction mixture solution, and the mixture was washed three times with 60 mL of dichloromethane. The organic layer solution was collected, dried over anhydrous magnesium sulfate, filtered and concentrated. The concentrated product was separated by silica gel column chromatography (hexanes: Et 2 O: CH 2 Cl 2, 1: 1: 1) to obtain an ester compound of formula 6 as a white solid (2 E , 2 ′ E ) -2,5-bis (3, 5-dimethoxy-4- (methoxymethoxy) phenyl) tetrahydrofuran-3,4-diyl) bis (methylene) bis (3- (4- (methoxymethoxy) phenyl) acrylate) (724 mg, 0.800 mmol, 72% yield) It was.

IR (neat): 2990 (w), 1760 (m), 1597 (m), 1509 (m), 1240 (s), 1146 (s), 970 (s), 832 (m), 757 (w) cm -One;

¹ H NMR (CDCl ₃ , 400 MHz): δ 7.61 (d, J = 15.9 Hz, 2H), 7.44 (d, J = 8.8 Hz, 4H), 7.03 (d, J = 8.8 Hz, 4H), 6.70 ( s, 4H), 6.28 (d, J = 15.9 Hz, 2H), 5.21 (s, 4H), 5.13 (s, 4H), 5.04-5.03 (m, 2H), 4.44-4.43 (m, 4H), 3.86 (s, 12H), 3.61 (s, 6H), 3.49 (s, 6H), 2.65-2.60 (m, 2H);

¹³ C NMR (CDCl ₃ , 100 MHz): δ 167.0, 159.2, 153.5, 145.2, 137.5, 133.8, 129.8, 127.8, 116.4, 115.1, 102.7, 98.1, 94.0, 83.0, 63.5, 57.1, 56.1, 56.0, 50.4;

HRMS (ESI-TOF) m / z: [M + Na] + Calcd for C 48 H 56 NaO 17 927.3415; Found 927.3416.

Production Example  4. Compound of formula (7) 2E, 2'E Synthesis of)-(-2,5-bis (4-hydroxy-3,5-dimethoxyphenyl) tetrahydrofuran-3,4-diyl) bis (methylene) bis (3- (4-hydroxyphenyl) acrylate)

Into a 25 mL round bottom flask was added a compound of formula 6 (212 mg, 0.234 mmol) introduced with a MOM-protecting group, methanol (7 mL) and 12 N HCl (0.8 mL) were added and stirred at 50 ° C. for 5 minutes. After concentrating the reaction solvent, the remaining reaction product was purified by silica gel column chromatography (EtOAc: CH2Cl2, 1: 1 ) to separate a yellow solid end product compound of formula (VII) with ((2 E, 2 'E ) - (- 2,5 -bis (4-hydroxy-3,5-dimethoxyphenyl) tetrahydrofuran-3,4-diyl) bis (methylene) bis (3- (4-hydroxyphenyl) acrylate) (119 mg, 0.163 mmol, 70% yield) was synthesized. The compound of formula 7 was named by the inventors 'NK11' or 'orizativol A'.

IR (neat): 3385 (br s), 2990 (w), 2363 (w), 1710 (m), 1597 (m), 1503 (m), 1233 (s), 1146 (s), 970 (s) 832 (m), 751 (w) cm < -1 >;

¹ H NMR (CD ₃ OD, 400 MHz): δ 7.39 (d, J = 15.9 Hz, 2H), 7.36 (d, J = 8.8 Hz, 4H), 6.80-6.74 (m, 8H), 6.21 (d, J = 15.9 Hz, 2H), 5.04 (d, J = 8.3 Hz, 2H), 4.45-4.37 (m, 4H), 3.82 (s, 12H), 2.70-2.63 (m, 2H);

¹³ C NMR (CD ₃ OD, 100 MHz): δ 169.1, 161.6, 149.6, 147.2, 136.7, 133.8, 131.5, 127.2, 117.1, 115.0, 105.2, 85.8, 64.8, 57.0, 51.8;

HRMS (ESI-TOF) m / z: [M + Na] + Calcd for C 40 H 40 NaO 13 751.2367; Found 751.2362.

Preparation Example 5 Synthesis of Compound of Formula 8

By applying the method of Preparation Examples 3 and 4, the trans coumarin acid derivative compound of formula 5 (the derivative of which the MOM-protecting group is bonded to the hydroxy group of 4-hydroxycinnamic acid) and the isomer of cis coumarin acid derivative (cis-4-hydroxycinnamic) The compound of Formula 8 was synthesized by binding a MOM-protecting group to a hydroxyl group of acid) to a tetrahydroxyfuran ring. The compound of Formula 8 was named by the inventors 'orizativol B'.

Preparation Example 6 Synthesis of Compound of Formula 9

By applying the method of Preparation Examples 3 and 4, instead of the trans coumarin acid derivative compound of Formula 5 (a derivative in which the MOM-protecting group is bonded to the hydroxyl group of 4-hydroxycinnamic acid), trans-cinnamic acid is added to Was synthesized.

Preparation Example 7 Synthesis of Compound of Formula 10

The compounds of the formula (10) were synthesized by binding the acetoxymethyl group to the tetrahydrofuran ring by applying the methods of Preparation Examples 3 and 4.

[Formula 10]

Production Example  8: compound of formula 11 (2,5- bis (3,5-dimethoxy-4- Synthesis of (methoxymethoxy) phenyl) tetrahydrofuran-3,4-diyl) diacetate)

The compound of Formula 11 was synthesized by applying a hydroxymethyl group to a tetrahydrofuran ring by applying the methods of Preparation Examples 3 and 4.

[Formula 11]

Experimental Example 1: Measurement of osteoblast differentiation activity and adipocyte differentiation inhibitory activity of NK11 compound

(1) Measurement of ALP (alkaline phosphatase) Promoting Activity of NK11 Compound

C3H10T1 / 2 cell lines derived from mouse embryonic fibroblasts are generally pluripotent stem cell lines capable of differentiating into various cell lineages including osteoblasts. One of the characteristics of osteoblasts is that it shows ALP (Alkaline phosphatase) activity, and thus the osteoblast differentiation effect was measured through ALP activity of C3H10T1 / 2 cell lines.

C3H10T1 / 2 cell line was incubated in DMEM medium containing 10% FBS, 1% penicillin and streptomycin at 37 ° C and 5% CO₂. The C3H10T1 / 2 cells were incubated with a medium containing 10 mM β-glycerophosphate and 50 μg / ml ascorbic acid for osteoblast differentiation at a concentration of 2.5 × 10 μs / ml in 6 well plates, and the NK11 compound was cultured. 0.1, 0.5, 1 and 5 μM, respectively, were added and differentiated for 9 days with changing medium every 3 days.

C3H10T1 / 2 pluripotent stem cell lines were cultured in DMEM medium containing 10% FBS, 1% penicillin and streptomycine at 37 ° C and 5% CO₂. The cells were incubated with a medium containing 10 mM β-glycerophosphate and 50 μg / ml ascorbic acid for osteoblast differentiation at a concentration of 2.5 × 10 cells / ml in 6 well plates, and the medium was changed every 3 days. Differentiation was carried out for 9 days with addition of 1 and 5 μM, respectively, followed by ALP staining using 5-Bromo-4-chloro-3-indolyl phosphate / nitro blue tetrazolium (BCIP / NBT).

After measuring the lab color space using the image file of the ALP staining well plate, NK11 compounds increased ALP production in a concentration-dependent manner as the treatment concentration increased compared to the negative control group treated with DMSO only. It can be seen (see Fig. 3).

(2) Determination of Adipocyte Differentiation Inhibitory Activity of NK11 Compound

After dispensing cells in 6-well plates in the same manner as in Experimental Example 1, for the adipocyte differentiation, 1 μM dexamethasone, 5 μg / mL insulin and 1 μM troglitazone, and 0.1, 0.5, 1, and 5 μM of NK11 compounds were respectively added. Incubated for 9 days. After the differentiation, the medium was removed and the cells were fixed with 10% neutral buffered formalin and stained with 0.5% oil red o solution. The results are shown in FIG. 4.

After measuring the lab color space using an image file of a well plate stained with an oil red o solution, NK11 compounds inhibited adipogenesis of adipocytes in a concentration-dependent manner compared to the negative control group treated with DMSO only. It confirmed the effect (refer FIG. 4).

(3) osteoblasts of NK11 compounds ( osteoblast ) Eruption factor  And fat cells Eruption factor  Expression level measurement

When the NK11 compound was treated, mRNA expression levels of osteoblast differentiation factor and adipocyte differentiation factor in cells were confirmed and shown in FIG. 6.

First, as in Experiment 1, C3H10T1 / 2 cell line was differentiated into osteoblasts, and the expression level of ALP, an osteoblast differentiation factor, was confirmed. In addition, the expression level of the adipocyte differentiation factors PPARγ, aP2, and CD36 were confirmed by differentiating C3H10T1 / 2 cell lines into adipocytes as in Experimental Example 2.

Total RNA extraction was done using TRIzol (Invitrogen). 1 μg of the isolated RNA was synthesized by cDNA by adding random primer, dNTP, and PrimeScript ™ Reverse Transcriptase (TaKaRa). The synthesized cDNA was subjected to realtime PCR with SYBR Premix Ex Taq (TaKaRa) using a primer.

The expression levels of adipocyte-related genes, CD36, aP2 and PPARγ, were reduced in a concentration-dependent manner, and the expression level of ALP, an important index related to bone formation, was high at 5 μM (see FIG. 5).

Experimental Example  2: Comparison of Osteoblast Differentiation and Adipocyte Differentiation Activity of NK11 Compound and Compound of Formula 8

The NK11 compound is a compound in which two trans coumarin acid derivative compounds are bonded to a carbon at positions 3 and 4 in a tetrahydrofuran ring, and the compound of Formula 8 is one of the trans coumarin acid derivatives in the furan ring of NK11. Isomers of NK11 compounds substituted with derivatives.

Osteoblast differentiation activity and adipocyte differentiation inhibitory activity of the compound of Formula 8 which is an isomer of NK11 compound were compared in the same manner as in Experimental Example 1.

(1) NK11 Compound and Compound of Formula 8 Alkaline phosphatase (ALP) Generation promotion activity comparison

As shown in the ALP staining pictures and graphs of Figure 6 it can be seen that the compound of formula 8 increases ALP production in a concentration-dependent manner, like the NK11 compound.

(2) Comparison of Adipocyte Differentiation Inhibitory Activity between NK11 Compound and Compound of Formula 8

As shown in the photographs and graphs of FIG. 7, the compound of Formula 8 showed an effect of inhibiting adipogenesis of adipocytes in a concentration-dependent manner, as in the NK11 compound, and the effect of each concentration was not substantially different from the NK11 compound.

(3) osteoblasts of the NK11 compound and the compound of the formula (8) osteoblast ) Eruption factor  And fat cell differentiation factor expression level comparison

As shown in the graph of FIG. 8, the compound of Formula 8 reduced the expression level of the adipocyte-related genes CD36, aP2, and PPARγ in a concentration-dependent manner, similar to the result of the NK11 compound of FIG. The expression level of was expressed high at 5 μM.

Experimental Example  2: Comparison of Osteoblast Differentiation and Adipocyte Differentiation Activity of NK11 Compound and Compounds of Formulas 4, 2, 10, 11, and 9

Compounds of formulas (4) and (11) have two hydroxymethyl groups attached to carbons at positions 3 and 4 in the tetrahydrofuran ring, except that compounds of formula (4) have positions 2 and 5 in the tetrahydrofuran ring. The hydroxymethyl of the carbon at position 4 of the phenyl group bonded to the carbon is bonded to the methoxymethyl protecting group, and the compound of the formula (11) is exposed as it is.

In addition, the compound of formula (2) is two methylcarboxy groups bonded to the carbon at positions 3 and 4 in the tetrahydrofuran ring, the compound of formula (10) is two at the carbon in positions 3 and 4 in the tetrahydrofuran ring It is a compound in which an acetoxymethyl group is bonded.

In addition, the compound of formula 9 is a compound in which cinnamic acid is bonded to the tetrahydrofuran ring instead of two transcoumarinic acid derivatives bonded to carbons at positions 3 and 4.

In Experimental Example 3, osteoblast differentiation activity and adipocyte differentiation inhibitory activity of the intermediate of NK11 compound synthesis process and NK11 compound-like structural compound were compared with NK11 compound in the same manner as in Experimental Example 1. The NK11 compound was treated at 1 μM and the remaining compounds were treated at 5 μM concentration.

(1) Comparison of ALP (alkaline phosphatase) production promoting activity

The intermediates of the synthesis of NK11 compounds and their analogous structural compounds of Formula 4, Formula 2, Formula 10 and Formula 11 were not different from those of the negative control group (DMSO) in the amount of ALP, and only the compound of Formula 9 was used as the drug of the NK11 compound. About 2 times the amount of ALP produced could be confirmed (see FIG. 9). Considering the concentrations of the NK11 compound and the compound of Formula 9 used in each experiment, the compound of Formula 9 was slightly lower than NK11 but had a significant ALP production promoting effect.

(2) Comparison of Adipocyte Differentiation Inhibitory Activity

Compounds of Formula 4, Formula 2, Formula 10, and Formula 11 did not differ from the negative control group (DMSO) in inhibiting adipocyte differentiation, and only the compound of Formula 9 was found to inhibit adipocyte differentiation with the NK11 compound (Fig. 10).

(2) Comparison of the expression levels of osteoblast differentiation factor and adipocyte differentiation factor

Compounds of Formula 4, Formula 2, Formula 10 and Formula 11 did not affect the expression level of the adipocyte-related genes CD36, aP2, PPARγ or the expression level of ALP, an important indicator related to bone formation (see FIG. 11).

However, the compound of Formula 9 decreased the expression level of adipocyte-related genes CD36, aP2, and PPARγ similarly to the result of NK11 compound, and the expression level of ALP, an important index related to bone formation, was high. However, the compound treated group of Formula 9 showed a higher inhibitory effect on the expression of adipocyte differentiation factor and the effect of promoting osteoblast differentiation factor in the NK11 compound treatment group even though the concentration of the compound was added 5 times higher than the NK11 compound.

Experimental Example 4: Animal test using ovariectomy bone metabolic disease animal model

To determine the effect of NK11 compounds on the treatment and prevention of bone metabolism diseases, the extracts were administered to ovarian resection mice, and then bone density was measured and histological analysis was performed.

(1) Animal breeding and ovariectomy

Eight week-old female SD (Sprague-Dawley) rats were purchased and had a one week accrual period. At 9 weeks of age, ovarian resection was performed and had a week of recovery. The samples were orally administered once daily for 8 wn, and the experimental animals were kept in one cage during the experiment period, and the environmental conditions were adjusted to room temperature 25 ㅁ 5 ℃, relative humidity 50 ㅁ 10%, and lighting time 12 hours. . Feed (AIN-93g) and water were freely consumed.

In the experimental group, the group was opened without cutting the ovary (Sham), the group administered with distilled water after ovarian resection (OVX), and the group receiving NK11 at the concentrations of 0.5 and 1 mg / kg after ovarian resection (NK11 0.5, NK11). 1) and 50 μg / kg of ESTRADIOL as a positive control and 50 mg / kg of non-glycoside soy isoflavone (ISOFLAVONE) were administered.

After 8 weeks of sample treatment, blood was anesthetized with diethyl ether, and the blood drawn from the blood was left standing for 30 minutes and centrifuged at 3000 rpm for 10 minutes. The analysis items measured triglyceride, total cholesterol, ALP concentration and GOT / GTP ratio.

After the experiment, the body weight of the sacrificed ovarian resected rats was measured, and the femoral bones were extracted and BMD was measured using a dual energy X-ray bone density analyzer (Norland pDEXA).

The femoral bones extracted from ovarian resection rats were rapidly frozen with liquid nitrogen and ground. Total RNA extraction was done using TRIzol (Invitrogen). 1 ug of isolated RNA was synthesized by cDNA by adding random primer, dNTP, PrimeScript ™ Reverse Transcriptase (TaKaRa). Synthesized cDNA was performed by realtime PCR with SYBR Premix Ex Taq (TaKaRa) using a primer to confirm the mRNA expression level of ALP, a differentiation factor related to osteoblast formation.

(2) weight measurement

Body weight increased in all the experimental groups in which ovarian resection was performed compared with the group opened without ovarian resection (Sham), but there was no significant difference (see FIG. 12).

(3) measurement of biochemical indicators

Significant differences in triglyceride, total cholesterol, ALP concentration and GOT / GTP ratio were not found in all experimental groups (see FIG. 13).

(4) BMD measurement

BMD of the ovarian ablation group (OVX) was significantly decreased compared to the non-ovarian ablation group (Sham). Compared with NK11 compound was found to increase bone density significantly in a concentration-dependent manner (see Figure 14).

(5) Measurement of osteoblast differentiation factor expression

ALP, a differentiation factor related to osteoblast formation from the femur, was significantly decreased in OVX group compared with ovarian resection group, compared with Sham. Rose to the enemy. NK11 compounds showed a tendency to increase the expression level of ALP in a concentration-dependent manner, and showed an expression level similar to that of the positive control at 1 mg / kg administration (see FIG. 15).

Hereinafter, a preparation example of a composition containing an extract of the present invention will be described, but the present invention is not intended to be limited thereto but only to be described in detail.

Formulation Example 1 Preparation of Powder

NK11 Compound 20 mg

Lactose 100 mg

Talc 10 mg

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

Formulation Example 2 Preparation of Tablet

NK11 Compound 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components and tableting according to the manufacturing method of the conventional tablet to prepare a tablet.

Formulation Example 3 Preparation of Capsule

NK11 Compound 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

NK11 Compound 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO _4, 12H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

NK11 Compound 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

After dissolving each component in purified water according to the usual method of preparing a liquid solution, adding lemon flavor appropriately, mixing the above components, adding purified water, adjusting the whole to 100 ml by adding purified water, and then filling into a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Health Functional Food

NK11 Compound 1,000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used in the manufacture of health functional food according to a conventional method.

Formulation Example 7 Preparation of Health Functional Food Beverage

NK11 Compound 1,000 mg

Citric Acid 1,000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components in accordance with the conventional dietary beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilized and then refrigerated It is used in the manufacture of the nutraceutical beverage composition of the present invention.

Although the composition ratio is a composition that is relatively suitable for the preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

Claims (16)

1. A composition for preventing or treating bone metabolic diseases comprising the compound of formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:

   [Formula 1]

   

   R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and R ₆ may be the same as or different from each other, and each independently hydrogen, hydroxy, alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, Any one selected from the group consisting of halogen and trifluoromethyl,

   R ₇ , R ₈ , R _9, and R ₁₀ may be the same as or different from each other, and each independently hydrogen or a phenyl group, and when R ₇ and R ₉ are hydrogen, R ₈ and R ₁₀ are phenyl, and R _{If 7} and R ₉ are phenyl then R ₈ and R ₁₀ are hydrogen and the phenyl is unsubstituted or substituted with any substituent selected from the group consisting of hydroxy, halogen and trifluoromethyl.
2. According to claim 1, wherein R ₁ , R ₃ , R ₄ and R ₆ may be the same or different from each other, each independently selected from the group consisting of alkyl having 1 to 6 carbon atoms and alkoxy having 1 to 6 carbon atoms Which one,

   The R ₂ and R ₅ may be the same or different from each other, each independently hydrogen, hydroxy, halogen and trifluoromethyl composition for preventing or treating bone metabolic disease, characterized in that any one selected from the group consisting of .
3. The method of claim 2, wherein the compound of Formula 1 is a compound of the following formula 7, Formula 8 or Formula 9 for preventing or treating bone metabolic disease:

   [Formula 7]

   

   [Formula 8]

   

   [Formula 9]

   

   .
4. 2. The composition for preventing or treating bone metabolic disease according to claim 1, which promotes osteoblast differentiation and inhibits adipocyte differentiation.
5. The composition for preventing or treating bone metabolic diseases according to claim 4, wherein the obesity and bone metabolic diseases are simultaneously prevented or treated.
6. A health functional food for improving or preventing bone metabolism diseases comprising the compound of Formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:

   [Formula 1]

   

   R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and R ₆ may be the same as or different from each other, and each independently hydrogen, hydroxy, alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, Any one selected from the group consisting of halogen and trifluoromethyl,

   R ₇ , R ₈ , R _9, and R ₁₀ may be the same as or different from each other, and each independently hydrogen or a phenyl group, and when R ₇ and R ₉ are hydrogen, R ₈ and R ₁₀ are phenyl, and R _{If 7} and R ₉ are phenyl then R ₈ and R ₁₀ are hydrogen and the phenyl is unsubstituted or substituted with any substituent selected from the group consisting of hydroxy, halogen and trifluoromethyl.
7. The method of claim 6,

   R ₁ , R ₃ , R ₄ and R ₆ may be the same or different from each other, and each independently one selected from the group consisting of alkyl having 1 to 6 carbon atoms and alkoxy having 1 to 6 carbon atoms,

   The R ₂ and R ₅ may be the same or different from each other, each independently hydrogen, hydroxy, halogen and trifluoromethyl, characterized in that any one selected from the group consisting of Nutraceutical.
8. The method of claim 7, wherein

   The compound of Formula 1 is a health functional food for improving or preventing bone metabolism disease, characterized in that the compound of Formula 7, Formula 8 or Formula:

   [Formula 7]

   

   [Formula 8]

   

   [Formula 9]

   

   .
9. The health functional food for improving or preventing bone metabolism disease according to claim 6, wherein osteoblast differentiation is promoted and adipocyte differentiation is suppressed.
10. 10. The health functional food for improving or preventing bone metabolism disease according to claim 9, wherein obesity and bone metabolic disease are simultaneously prevented or treated.
11. A compound having the activity of promoting osteoblast differentiation of Formula 1 and inhibiting adipocyte differentiation:

    [Formula 1]

    

    R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and R ₆ may be the same as or different from each other, and each independently hydrogen, hydroxy, alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, Any one selected from the group consisting of halogen and trifluoromethyl,

    R ₇ , R ₈ , R _9, and R ₁₀ may be the same as or different from each other, and each independently hydrogen or a phenyl group, and when R ₇ and R ₉ are hydrogen, R ₈ and R ₁₀ are phenyl, and R _{If 7} and R ₉ are phenyl then R ₈ and R ₁₀ are hydrogen and the phenyl is unsubstituted or substituted with any substituent selected from the group consisting of hydroxy, halogen and trifluoromethyl.
12. The method according to claim 11, wherein R ₁ , R ₃ , R ₄ and R ₆ may be the same or different from each other, each independently selected from the group consisting of alkyl having 1 to 6 carbon atoms and alkoxy having 1 to 6 carbon atoms. Which one,

    R ₂ and R ₅ may be the same as or different from each other, and each independently promotes osteoblast differentiation, characterized in that any one selected from the group consisting of hydrogen, hydroxy, halogen and trifluoromethyl fat Compounds having activity that inhibits cell differentiation.
13. According to claim 12, wherein the compound of Formula 1 is a compound having the activity of promoting osteoblast differentiation and inhibiting adipocyte differentiation, characterized in that the compound of Formula 7, Formula 8 or Formula 9:

    [Formula 7]

    

    [Formula 8]

    

    [Formula 9]

    

    .
14. Preparing a compound of Chemical Formula 6 by reacting a compound of Chemical Formula 4 with a compound of Chemical Formula 5; And removing the MOM-protecting group from the compound of Chemical Formula 6; a method of preparing a compound having an activity of promoting osteoblast differentiation of Chemical Formula 7 and inhibiting adipocyte differentiation.

    [Formula 4]

    

    [Formula 5]

    

    [Formula 6]

    

    [Formula 7]

    

    .
15. The method of claim 14, further comprising: reacting a compound of Formula 2 with chloromethylmethyl ether to prepare a compound of Formula 3; And reacting the compound of Formula 3 with diisobutylaluminum hydride to prepare the compound of Formula 4; Method for preparing a compound having:

    [Formula 2]

    

    [Formula 3]

    

    .
16. The method of claim 15, wherein the compound of Formula 2 promotes osteoblast differentiation of formula (7) characterized in that it is synthesized from 4-hydroxy-3,5-dimethoxycinnamic acid and has the activity of inhibiting adipocyte differentiation Method for preparing the compound.

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