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WO2017209410A1 - Nouveau composé promoteur de différenciation d'ostéoblastes et inhibiteur de différenciation d'adipocytes, son procédé de préparation et son application - Google Patents

Nouveau composé promoteur de différenciation d'ostéoblastes et inhibiteur de différenciation d'adipocytes, son procédé de préparation et son application Download PDF

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Publication number
WO2017209410A1
WO2017209410A1 PCT/KR2017/004972 KR2017004972W WO2017209410A1 WO 2017209410 A1 WO2017209410 A1 WO 2017209410A1 KR 2017004972 W KR2017004972 W KR 2017004972W WO 2017209410 A1 WO2017209410 A1 WO 2017209410A1
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Prior art keywords
formula
compound
group
differentiation
phenyl
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Korean (ko)
Inventor
김현석
박기문
방민혁
양희진
이윤미
김기현
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SUNG KYUN BIOTECH CO Ltd
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SUNG KYUN BIOTECH CO Ltd
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Priority to JP2018560108A priority Critical patent/JP6605760B2/ja
Priority to US16/300,086 priority patent/US10494356B2/en
Priority to CN201780029851.0A priority patent/CN109152762B/zh
Priority claimed from KR1020170059133A external-priority patent/KR101897726B1/ko
Publication of WO2017209410A1 publication Critical patent/WO2017209410A1/fr
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide

Definitions

  • 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.
  • substances such as calcium and other essential minerals in the body, such as phosphorus or magnesium.
  • Bone is composed of osteoblasts (osteoblast), osteocytes (osteocyte), osteoclasts (osteoclast).
  • 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.
  • 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- ⁇ B ligand (RANKL) and osteoprotegerin (OPG). Keep it.
  • RNKL nuclear factor- ⁇ B ligand
  • OPG osteoprotegerin
  • 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.
  • T-score standard deviation from the average adult's average bone mass
  • 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.
  • 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.
  • 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.
  • 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).
  • 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.
  • 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.
  • 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.
  • 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.
  • the compound having the activity of promoting osteoblast differentiation and inhibiting adipocyte differentiation of the present invention is a compound of Formula 1 below.
  • 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.
  • 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.
  • the compound of Formula 1 is a compound of Formula 7, Formula 8, or Formula 9.
  • 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.
  • the reaction 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • BMD bone mineral density
  • 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.
  • a drug 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • the beverage does not have any particular limitation, and may contain various flavors or natural carbohydrates as additional components, as in general beverages.
  • 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.
  • natural flavoring agents such as, 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.
  • 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.
  • 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 PPAR ⁇ , 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.
  • 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 .
  • ALP alkaline phosphatase
  • 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 PPAR ⁇ , aP2, CD36, ALP in the C3H10T1 / 2 cell line according to the NK11 compound treatment in Experimental Example 1.
  • ALP alkaline phosphatase
  • FIG. 8 is a graph showing changes in the expression level of mRNA PPAR ⁇ , aP2, CD36, ALP in the C3H10T1 / 2 cell line according to the compound of Formula 8 in Experimental Example 2.
  • 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 PPAR ⁇ , 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.
  • FIG. 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. 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.
  • the compound of formula (II) was prepared using a known synthesis method from 4-hydroxy-3,5-dimethoxycinnamic acid.
  • 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 ⁇ L, 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.
  • DIBAL-H diisobutylaluminum hydride
  • 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 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'.
  • trans-cinnamic acid is added to Was synthesized.
  • 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.
  • 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.
  • 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.
  • ALP Alkaline phosphatase
  • 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 ⁇ -glycerophosphate and 50 ⁇ g / ml ascorbic acid for osteoblast differentiation at a concentration of 2.5 ⁇ 10 ⁇ s / ml in 6 well plates, and the NK11 compound was cultured. 0.1, 0.5, 1 and 5 ⁇ M, 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 ⁇ -glycerophosphate and 50 ⁇ g / 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 ⁇ M, respectively, followed by ALP staining using 5-Bromo-4-chloro-3-indolyl phosphate / nitro blue tetrazolium (BCIP / NBT).
  • BCIP / NBT 5-Bromo-4-chloro-3-indolyl phosphate / nitro blue tetrazolium
  • NK11 compounds 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).
  • NK11 compounds 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).
  • osteoblasts of NK11 compounds ( osteoblast ) Eruption factor And fat cells Eruption factor Expression level measurement
  • NK11 compound 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.
  • C3H10T1 / 2 cell line was differentiated into osteoblasts, and the expression level of ALP, an osteoblast differentiation factor, was confirmed.
  • the expression level of the adipocyte differentiation factors PPAR ⁇ , aP2, and CD36 were confirmed by differentiating C3H10T1 / 2 cell lines into adipocytes as in Experimental Example 2.
  • RNA extraction was done using TRIzol (Invitrogen). 1 ⁇ g of the isolated RNA was synthesized by cDNA by adding random primer, dNTP, and PrimeScript TM Reverse Transcriptase (TaKaRa). The synthesized cDNA was subjected to realtime PCR with SYBR Premix Ex Taq (TaKaRa) using a primer.
  • adipocyte-related genes CD36, aP2 and PPAR ⁇
  • ALP an important index related to bone formation
  • 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.
  • the compound of formula 8 increases ALP production in a concentration-dependent manner, like the NK11 compound.
  • 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.
  • osteoblasts of the NK11 compound and the compound of the formula (8) osteoblast Eruption factor And fat cell differentiation factor expression level comparison
  • the compound of Formula 8 reduced the expression level of the adipocyte-related genes CD36, aP2, and PPAR ⁇ in a concentration-dependent manner, similar to the result of the NK11 compound of FIG.
  • the expression level of was expressed high at 5 ⁇ M.
  • 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.
  • 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.
  • 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.
  • 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.
  • Compounds of Formula 4, Formula 2, Formula 10 and Formula 11 did not affect the expression level of the adipocyte-related genes CD36, aP2, PPAR ⁇ or the expression level of ALP, an important indicator related to bone formation (see FIG. 11).
  • the compound of Formula 9 decreased the expression level of adipocyte-related genes CD36, aP2, and PPAR ⁇ similarly to the result of NK11 compound, and the expression level of ALP, an important index related to bone formation, was high.
  • 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.
  • NK11 compounds were administered to ovarian resection mice, and then bone density was measured and histological analysis was performed.
  • 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 ⁇ g / kg of ESTRADIOL as a positive control and 50 mg / kg of non-glycoside soy isoflavone (ISOFLAVONE) were administered.
  • 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).
  • RNA extraction was done using TRIzol (Invitrogen). 1 ug of isolated RNA was synthesized by cDNA by adding random primer, dNTP, PrimeScript TM 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.
  • ALP a differentiation factor related to osteoblast formation from the femur
  • 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).
  • the above ingredients are mixed and filled in an airtight cloth to prepare a powder.
  • the above ingredients are mixed and filled into gelatin capsules to prepare capsules.
  • the amount of the above ingredient is prepared per ampoule (2 ml).
  • Vitamin B6 0.5 mg
  • 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.
  • 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.
  • composition ratio is a composition that is relatively suitable for the preferred beverage in a preferred embodiment
  • compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

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Abstract

La présente invention concerne un composé ayant l'activité de stimulation de la différenciation d'ostéoblastes et d'inhibition de la différenciation d'adipocytes, et un procédé de préparation associé. Un nouveau composé de la présente invention augmente l'expression du gène ALP impliqué dans la différenciation d'ostéoblastes, régule l'expression des gènes PPARγ, aP2 et CD36 impliqués dans la différenciation d'adipocytes, augmente une densité de masse osseuse (BMD) dans un modèle d'animal ostéoporotique ovariectomisé, et diminue un taux d'adipocytes dans la moelle osseuse. Par conséquent, le composé peut être utilisé en tant que composant efficace de médicaments ou d'aliments fonctionnels de santé pour des maladies métaboliques osseuses ou l'obésité.
PCT/KR2017/004972 2016-05-31 2017-05-12 Nouveau composé promoteur de différenciation d'ostéoblastes et inhibiteur de différenciation d'adipocytes, son procédé de préparation et son application Ceased WO2017209410A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
JP2018560108A JP6605760B2 (ja) 2016-05-31 2017-05-12 造骨細胞分化を促進するとともに脂肪細胞分化を抑制する新規化合物、その製造方法及びその応用
US16/300,086 US10494356B2 (en) 2016-05-31 2017-05-12 Compound promoting osteoblast differentiation and inhibiting adipocyte differentiation, preparation method thereof and application thereof
CN201780029851.0A CN109152762B (zh) 2016-05-31 2017-05-12 促进成骨细胞分化和抑制脂肪细胞分化的化合物、其制备方法及其应用

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2016-0067570 2016-05-31
KR20160067570 2016-05-31
KR1020170059133A KR101897726B1 (ko) 2016-05-31 2017-05-12 조골세포 분화를 촉진하며 지방세포 분화를 억제하는 신규 화합물, 그 제조방법 및 그 응용
KR10-2017-0059133 2017-05-12

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WO2017209410A1 true WO2017209410A1 (fr) 2017-12-07

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PCT/KR2017/004972 Ceased WO2017209410A1 (fr) 2016-05-31 2017-05-12 Nouveau composé promoteur de différenciation d'ostéoblastes et inhibiteur de différenciation d'adipocytes, son procédé de préparation et son application

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WO (1) WO2017209410A1 (fr)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150055667A (ko) * 2013-11-13 2015-05-22 주식회사 케이스템셀 사철쑥 유래 스코파론 함유 추출물을 유효성분으로 포함하는 골다공증 예방 또는 치료용 조성물

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KR20150055667A (ko) * 2013-11-13 2015-05-22 주식회사 케이스템셀 사철쑥 유래 스코파론 함유 추출물을 유효성분으로 포함하는 골다공증 예방 또는 치료용 조성물

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