KR20230116485A - Composition for promoting differentiation of beige fat and treating metabolic diseases containing TUDCA as an active ingredient - Google Patents

Abstract

The present invention relates to a composition containing TUDCA as an active ingredient, for promoting the differentiation of beige fat cells and preventing or treating metabolic diseases. The present invention confirmed that endoplasmic reticulum stress inhibits the differentiation of beige fat cells, and found for the first time that endoplasmic reticulum stress reduces the expression of a thermogenic complex related to the differentiation of beige fat cells. Accordingly, it is confirmed that when fat cells in which endoplasmic reticulum stress is induced are treated with TUDCA, which reduces endoplasmic reticulum stress, the expression of factors in the UPR pathway, which is an endoplasmic reticulum stress response, was significantly decreased, and that the expression of factors related to UCP1 and thermogenic complexes was increased, thereby promoting differentiation of beige fat cells. In addition, it was confirmed that in high-fat diet mice treated with TUDCA of the present invention, body weight was reduced, metabolic abnormalities were improved, beige fat cells in adipose tissue increased, and the size and density of white fat tissue decreased, and the improvement effect of metabolic diseases induced by endoplasmic reticulum stress was confirmed. In addition, it was confirmed that the differentiation of beige fat cells can be promoted more effectively when treated with TUDCA before having obesity.

Description

Composition for promoting differentiation of beige fat and treating metabolic diseases containing TUDCA as an active ingredient}

The present invention relates to a composition for promoting differentiation of beige fat and treating metabolic diseases, comprising TUDCA as an active ingredient.

As the obesity problem becomes more serious, the perception that obesity itself is a disease is spreading. Whether or not obesity progresses can be typically determined by overweight, and can be specifically determined by an increase in body fat in a specific region (eg, abdominal obesity, etc.). When classified as obese, that is, obese people have higher morbidity and mortality than other diseases, and compared to people with normal weight, the death rate due to diabetes is 4 times, the death rate due to mild liver disease is 2 times, and the death rate due to cerebrovascular disease is 4 times higher. It is reported that the mortality rate is 1.6 times higher and the mortality rate due to coronary artery disease is about 1.8 times higher.

Obesity is a progressive metabolic disorder caused by an imbalance in whole-body energy homeostasis. Excessive intake of energy compared to consumed energy causes energy imbalance and increases white adipose tissue, which is a major cause of metabolic diseases such as obesity and diabetes.

On the other hand, beige/brite adipocytes that express uncoupling protein 1 (UCP1) and are rich in mitochondria are differentiated into beige fat precursor cells or converted from white adipocytes, resulting in inguinal white fat. It develops within tissue (inguinal white adipose tissue, iWAT) depots. Beige adipocytes are specialized in consuming excess energy through UCP1-mediated thermogenesis, and thus have different characteristics from white adipose tissue responsible for energy storage. In humans, a high level of beige fat activity correlates with a lean body type, and an important role of beige fat in human metabolism has been reported. Due to the unique function of beige fat, it can be used as a therapeutic target to activate beige fat and increase energy consumption. As such, transcription factors and physiologically active molecules that promote the development of beige fat cells have potential as ideal anti-obesity drugs.

On the other hand, the endoplasmic reticulum (ER) is a cellular organelle responsible for the folding of newly synthesized polypeptides, lipid biosynthesis, and calcium storage. Due to its physiological importance, maintaining ER homeostasis when exposed to various stimuli is essential for cellular function and It is known to be important for cell survival. When endoplasmic reticulum homeostasis is disrupted, collectively referred to as endoplasmic reticulum stress, it has been reported that a protective response mechanism known as the unfolded protein response (UPR) is induced. UPR is initiated by activation of three ER membrane proteins: PKR-like ER kinase (PERK), inositol-requiring enzyme 1α (IRE1α), and activating transcription factor 6α (ATF6α). (eIF2α) phosphorylation and C/EBP homologous protein (CHOP) expression are increased. It has been reported that endoplasmic reticulum stress is related to the differentiation and function of white adipocytes. For example, phosphorylated eIF2α and subsequent induction of CHOP protein lead to inhibition of adipogenesis, whereas the IRE1α-Xbp1 pathway and ATF6 have been reported to promote white adipocyte differentiation, respectively. endoplasmic reticulum Stressed white adipocytes show functional disorders including impaired insulin signal transduction and dysregulation of adipokine secretion, but it has not been reported whether endoplasmic reticulum stress affects beige adipocyte differentiation.

An object of the present invention is to provide a pharmaceutical composition for preventing or treating metabolic diseases, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient. is to do

Another object of the present invention is to provide a food composition for preventing or improving metabolic diseases, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient. is to do

Another object of the present invention is to provide a composition for promoting differentiation of beige adipocytes, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient. .

Another object of the present invention is to inhibit the differentiation and thermogenesis of beige adipocytes cells, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient. caused by inhibition of complex expression. To provide a pharmaceutical composition for preventing or treating metabolic diseases.

In order to achieve the above object, the present invention, the prevention or treatment of metabolic diseases comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient It provides a pharmaceutical composition for use.

In addition, the present invention provides a food composition for preventing or improving metabolic diseases, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient. .

In addition, the present invention provides a composition for promoting differentiation of beige adipocytes, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention is a transcription complex inhibiting the differentiation of beige adipocytes and generating thermogenesis, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient. Provided is a pharmaceutical composition for preventing or treating metabolic diseases caused by expression inhibition.

The present invention first identified that endoplasmic reticulum stress suppresses the differentiation of beige adipocytes and thereby reduces the expression of a thermogenic complex including UCP1 protein. Accordingly, when TUDCA, which reduces endoplasmic reticulum stress, was treated with endoplasmic reticulum stress-induced preadipocytes, the expression of factors of the UPR pathway, which is an endoplasmic reticulum stress response, was significantly reduced, and the expression of factors related to UCP1 and the thermogenic complex was increased. increased, it was confirmed that the differentiation of beige adipocytes was promoted. In addition, in mice fed a high-fat diet treated with the TUDCA of the present invention, it was confirmed that body weight was decreased, metabolic abnormalities were improved, and beige fat cells in adipose tissue increased and the size and density of white adipose tissue decreased, The improvement effect of metabolic diseases induced by endoplasmic reticulum stress was confirmed. In addition, it was confirmed that the differentiation of beige adipocytes can be more effectively promoted when TUDCA is treated before obesity is generated, and thus it can be usefully used in related industries.

1 is a diagram confirming that differentiation of beige adipocytes is inhibited by the induction of endoplasmic reticulum stress (A: Oil Red O staining results and quantification of staining results, B: confirmation of UCP1 expression changes induced by endoplasmic reticulum stress, C: endoplasmic reticulum stress-related genes). and protein expression level of beige fat differentiation factor, D: overexpression of CHOP protein through adenovirus, E: quantification of change in mRNA expression level of beige fat cell differentiation factor due to overexpression of CHOP protein, F: CHOP protein Confirmation of changes in protein expression levels of fat differentiation factors due to overexpression).
Figure 2 is a diagram confirming the differentiation of beige adipocytes according to treatment with an endoplasmic reticulum stress reducing agent (A: change in expression pattern of marker genes for differentiation of beige fat by TUDCA treatment, B: expression pattern of UCP1 and CHOP protein according to TUDCA treatment). Confirmation of changes, C: Confirmation of changes in beige fat differentiation according to treatment with 4-PBA, another endoplasmic reticulum stress reliever, D: Confirmation of UCP1 protein expression according to 4-PBA treatment, E: Differentiation of beige fat according to 4-PBA treatment confirmation of related factor protein expression).
Figure 3 is a diagram confirming the effect of beige fat differentiation according to the treatment of TUDCA in the early stage of beige adipocyte differentiation (A: Confirmation of endoplasmic reticulum stress-related gene expression patterns according to the differentiation induction period, B: Beige according to the differentiation induction period) Confirmation of changes in protein amount of factors related to color fat differentiation, C: Confirmation of changes in the differentiation of beige adipocytes according to the date of TUCDA treatment during the period of beige fat differentiation, D: Differentiation of beige adipocytes according to the date of TUCDA treatment during the period of beige fat differentiation Check changes in protein expression of related genes).
Figure 4 is a diagram confirming the differentiation of beige fat according to the treatment of TUDCA in mice consuming a high-fat diet (A: confirmation of mouse weight change after TUDCA administration, B: confirmation of food intake after TUDCA administration, C: mouse fat and Check liver tissue size, D: check histological changes by tissue (liver, visceral fat, inguinal fat, brown adipose tissue) after TUDCA administration, E: check fasting blood sugar after TUDCA administration, F: check glucose resistance after TUDCA administration, G: Insulin resistance confirmed after TUDCA administration, H: UCP1 and CHOP gene expression confirmed after TUDCA administration, I: Protein expression of fat differentiation and endoplasmic reticulum stress-related genes confirmed after TUDCA administration).
Figure 5 is a diagram confirming the change in beige fat differentiation according to TUDCA pretreatment before ingestion of a high-fat diet (A: confirmation of mouse weight change after TUDCA administration, B: confirmation of mouse fat and liver tissue size after TUDCA administration, C: TUDCA administration Confirm histological changes by tissue (liver, visceral fat, inguinal fat, brown adipose tissue), D: Check fasting blood sugar after TUDCA administration, E: Check glucose resistance after TUDCA administration, F: Check insulin resistance after TUDCA administration, G: Rectal temperature change after TUDCA administration, H: UCP1-positive cells confirmed and quantified in adipose tissue after TUDCA administration, I: Adipocyte size and density change in adipose tissue after TUDCA administration, J: UCP1 gene expression in adipose tissue after TUDCA administration Confirmation, K: expression of proteins related to endoplasmic reticulum stress and adipocyte differentiation after administration of TUDCA).

The present invention provides a pharmaceutical composition for preventing or treating metabolic diseases, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient.

As used herein, the term "prevention" refers to any action that suppresses symptoms or delays the progression of a specific disease by administering the composition of the present invention.

The term "treatment" used in the present invention refers to all activities that improve or beneficially change the symptoms of a specific disease by administering the composition of the present invention.

The "beige adipocytes" of the present invention are heat-generating fat cells that exist in white fat to maintain body temperature from a low external temperature, have a large number of mitochondria in the cells, and Adipocytes that express developmental regulatory proteins and form multifocal fat. Beige adipocytes exhibit different characteristics from white adipocytes, and while white adipocytes are cells that store energy, beige adipocytes are adipocytes that consume energy to generate heat. Beige fat has been reported to be produced from white fat cells through a path called browning, and has different characteristics from white fat cells, so it is attracting attention as a treatment target for various metabolic diseases including obesity.

The "endoplasmic reticulum stress" of the present invention is a protective reaction mechanism known as the unfolded protein response (UPR), and the UPR is PKR-like ER kinase (PERK), Inositol-requiring enzyme 1α (IRE1α), Activating Transcription Factor 6α (ATF6α), etc. It is initiated by activation of three ER transmembrane proteins, and continuous activation of PERK results in phosphorylation of eukaryotic initiation factor 2 alpha (eIF2α) and C/EBP homologous protein (CHOP) refers to a series of reactions that induce the expression of

In addition, it is known that the induction of phosphorylated eIF2α and CHOP induces inhibition of adipogenesis, and the IREα-Xbp1 pathway and ATF6 each promote white adipocyte differentiation. For example, CHOP is known to suppress white adipogenesis by forming a complex with C/EBPβ to block transcription of target genes responsible for white adipose differentiation. In the present invention, it was confirmed that the differentiation of beige adipocytes was also suppressed due to the induction of phosphorylation of eIF2a and the expression of CHOP protein by endoplasmic reticulum stress, similar to the differentiation of white adipocytes. Based on these results, it was confirmed that treatment with TUDCA, which relieves endoplasmic reticulum stress, promotes differentiation of beige adipose tissue due to a decrease in eIF2a phosphorylation and a decrease in CHOP protein expression.

In the present invention, it was confirmed that TUDCA increases the formation of a transcriptional complex associated with activation of thermogenic programming, which is a major feature of beige adipocyte differentiation.

In the transcription complex involved in the thermogenic programming activation of the present invention, PRDM16 is a key regulator for the physical interaction of the essential transcription factors PPARγ, C/EBPβ, PPARα and PGC-1α. In the present invention, it was confirmed that the expression of thermogenic genes including UCP1, Cidea, Pparγ, and PGC-1α, as well as Prdm16 and C/EBPα were significantly suppressed by the introduction of CHOP, and treatment with TUDCA inhibited the generation of beige adipocytes. and increased expression of UCP1, Cidea, Prdm16, PGC-1α and C/EBPa. Based on this, TUDCA relieves endoplasmic reticulum stress and suppresses the eIF2α/CHOP pathway, It was confirmed that the expression of genes related to thermogenic programming was increased, and as a result, differentiation of beige adipocytes was promoted.

"Tauroursodeoxycholic acid (TUDCA)" of the present invention may be a compound represented by Formula 1 below.

The pharmaceutically acceptable salt may include an acid addition salt formed by a pharmaceutically acceptable free acid, and organic acids and inorganic acids may be used as the free acid. The organic acids include citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, metasulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, glutamic acid, aspartic acid, and the like. It may include, but is not limited to. In addition, the inorganic acid may include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, and the like, but is not limited thereto.

The pharmaceutical composition of the present invention may further include an adjuvant in addition to the active ingredient. As long as the adjuvant is known in the art, any one may be used without limitation, but, for example, Freund's complete adjuvant or incomplete adjuvant may be further included to increase the effect.

The pharmaceutical composition according to the present invention may be prepared in the form of incorporating the active ingredient into a pharmaceutically acceptable carrier. Here, the pharmaceutically acceptable carrier includes carriers, excipients and diluents commonly used in the pharmaceutical field. Pharmaceutically acceptable carriers usable in the pharmaceutical composition of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical composition of the present invention may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories or sterile injection solutions according to conventional methods, respectively. .

When formulated, it may be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations contain at least one or more excipients such as starch, calcium carbonate, sucrose, lactose, and gelatin in addition to active ingredients. It can be prepared by mixing etc. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions for oral administration, emulsions, syrups, etc. In addition to commonly used diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. can Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for suppositories, witepsol, tween 61, cacao paper, laurin paper, glycerogelatin, and the like may be used.

The pharmaceutical composition according to the present invention can be administered to a subject by various routes. All modes of administration are contemplated, eg oral, intravenous, intramuscular, subcutaneous, intraperitoneal injection.

The dosage of the pharmaceutical composition according to the present invention is selected in consideration of the age, weight, sex, and physical condition of the subject. It is obvious that the concentration of the active ingredient included in the pharmaceutical composition can be variously selected according to the subject, and is preferably included in the pharmaceutical composition at a concentration of 0.01 to 5,000 μg/ml. If the concentration is less than 0.01 μg/ml, pharmacological activity may not appear, and if the concentration exceeds 5,000 μg/ml, toxicity to the human body may be exhibited.

According to one embodiment of the present invention, the metabolic disease may be induced by endoplasmic reticulum stress (ER stress).

According to one embodiment of the present invention, the composition may increase the formation of beige adipocytes.

According to one embodiment of the present invention, formation of the beige adipocytes may be induced by expression of a thermogenic transcription complex.

According to one embodiment of the present invention, the composition may increase the expression of a thermogenic transcription complex, wherein the thermogenic transcription complex comprises uncoupling protein 1 (UCP1), cytochrome c oxidase subunit 8B (COX8b), PR domain containing 16 (PRDM16), Peroxisome proliferator-activated receptor gamma (PPARγ), Peroxisome proliferator-activated receptor gamma alpha (PPARα), Ccaat-enhancer-binding proteins α (C/EBPα), Ccaat-enhancer-binding proteins β (C/ EBPβ), a factor selected from the group consisting of Peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC-1α), Perilipin, Voltage-dependent anion-selective channel 1 (VDAC1), and Cell Death Inducing DFFA Like Effector A (CIDEA) may be

According to one embodiment of the present invention, the composition may inhibit the expression of an endoplasmic reticulum stress factor, the endoplasmic reticulum stress may induce an unfolded protein response (UPR), and the endoplasmic reticulum stress factor PKR-like ER kinase (PERK), inositol-requiring enzyme 1α (IRE1α), activating transcription factor 6α (ATF6α), activating transcription factor 4 (ATF4), eukaryotic initiation factor 2α (eIF2α) and C/EBP homologous protein (CHOP ) It may be a factor selected from the group consisting of.

According to one embodiment of the present invention, the composition may reduce body weight.

According to one embodiment of the present invention, the composition may reduce the size or density of adipocytes in tissues, and the tissues include inguinal white adipose tissue (iWAT), epididymal white adipose tissue (epididymal It may be white adipose tissue (eWAT) or liver tissue.

According to one embodiment of the present invention, the composition may inhibit metabolic disorders induced by metabolic diseases, and the metabolic abnormalities may be increased glucose resistance or insulin resistance.

According to one embodiment of the present invention, the composition may reduce fasting blood sugar.

According to one embodiment of the present invention, the metabolic disease is obesity, type 1 diabetes, type 2 diabetes, hypoglycemia, hypercholesterinemia, It may be selected from the group consisting of hyperlipidemia, hemochromatosis, amyloidosis and porphyria, preferably obesity, but is not limited thereto.

In addition, the present invention provides a food composition for preventing or improving metabolic diseases, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient. .

As used herein, the term "improvement" refers to any action that at least reduces the severity of a parameter related to the condition being treated, for example, a symptom.

In addition to containing the active ingredient of the present invention, the food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients like conventional food compositions.

Examples of the aforementioned natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrins, cyclodextrins, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agents described above, natural flavoring agents (thaumatin), stevia extracts (eg rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can advantageously be used. The food composition of the present invention can be formulated in the same way as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gum, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements, etc. there is

In addition, the food composition, in addition to the active ingredient extract, various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, Alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like may be contained. In addition, the food composition of the present invention may contain fruit flesh for preparing natural fruit juice, fruit juice beverages, and vegetable beverages.

The functional food composition of the present invention may be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of preventing or treating metabolic diseases. In the present invention, 'health functional food composition' refers to a food manufactured and processed using raw materials or ingredients having useful functionality for the human body according to Health Functional Food Act No. 6727, and the structure and function of the human body It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological functions. The health functional food of the present invention may contain ordinary food additives, and the suitability as a food additive is determined according to the general rules of the Food Additive Code and General Test Methods approved by the Food and Drug Administration, unless otherwise specified. It is judged according to standards and standards. Examples of the items listed in the 'Food Additive Code' include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, kaoliang pigment, and guar gum; and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations. For example, a health functional food in the form of a tablet is obtained by granulating a mixture obtained by mixing the active ingredient of the present invention with an excipient, a binder, a disintegrant, and other additives in a conventional manner, and then adding a lubricant or the like to compression molding, or as described above. The mixture can be directly compression molded. In addition, the health functional food in the form of a tablet may contain a flavoring agent and the like as needed. Among health functional foods in the form of capsules, hard capsules can be prepared by filling a mixture in which the active ingredient of the present invention is mixed with additives such as excipients in a normal hard capsule. It can be prepared by filling the mixture mixed with gelatin in a capsule base. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary. The health functional food in the form of a pill can be prepared by molding a mixture of the active ingredient of the present invention mixed with an excipient, a binder, a disintegrant, etc. by a conventionally known method, and can be coated with sucrose or other coating agent if necessary, Alternatively, the surface may be coated with a material such as starch or talc. Health functional food in the form of granules can be prepared in granular form by a conventionally known method of mixing the active ingredient of the present invention with excipients, binders, disintegrants, etc., and, if necessary, flavoring agents, flavoring agents, etc. can

In addition, the present invention provides a composition for promoting differentiation of beige adipocytes, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient.

According to one embodiment of the present invention, the composition contains intracellular uncoupling protein 1 (UCP1), Cytochrome c oxidase subunit 8B (COX8b), PR domain containing 16 (PRDM16), Peroxisome proliferator-activated receptor gamma (PPARγ), Peroxisome proliferator-activated receptor gamma alpha (PPARα), Ccaat-enhancer-binding proteins α (C/EBPα), Ccaat-enhancer-binding proteins β (C/EBPβ), Peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC- 1α), Perilipin, Voltage-dependent anion-selective channel 1 (VDAC1), and Cell Death Inducing DFFA Like Effector A (CIDEA).

According to one embodiment of the present invention, the composition, intracellular PKR-like ER kinase (PERK), inositol-requiring enzyme 1α (IRE1α), activating transcription factor 6α (ATF6α), activating transcription factor 4 (ATF4), eukaryotic It may be to reduce the expression of a factor selected from the group consisting of initiation factor 2α (eIF2α) and C/EBP homologous protein (CHOP).

According to one embodiment of the present invention, the differentiation is dexamethasone, 3-isobutyl-1-methylxanthine, insulin, indomethacin, 3 It may be derived from a compound selected from the group consisting of 3'-triiodo-L-thyronine (3,3 ',5-triiodo-L-thyronine) and rosiglitazone.

In addition, the present invention, containing tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient, inhibits the differentiation of beige adipocytes cells, and generates heat. Provided is a pharmaceutical composition for preventing or treating metabolic diseases in which expression of a transcription complex is suppressed.

Hereinafter, the present invention will be described in more detail by examples. These examples are merely for explaining the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited to these examples.

<Preparation Example 1> Adipocyte culture

In order to confirm the effect of promoting the differentiation of beige fat of tauroursodeoxycholic acid (TUDCA) of the present invention, adipocytes were cultured. Specifically, adipose tissue was isolated from the inguinal region of the mouse, and at 37° C. for 20 to 30 minutes, 2 ml collagenase I buffer (250 U/g Lakewood, NJ, USA and 30 mg/mL bovine serum). Homogenates were obtained by digestion in Dulbecco's Modified Eagle's medium containing albumin). The obtained homogenate was filtered through a 100 μm cell strainer and centrifuged at 1200×g for 5 minutes to remove the supernatant. Isolated stromal-vascular cells (SVC) were cultured in Dulbecco's modified Eagle's medium (DMEM; Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. It was resuspended in 37 ℃, 5% CO ₂ It was cultured under conditions. For the differentiation of cultured SVC into beige adipocytes, 2 μg/mL dexamethasone, 0.5 mmol/L 3-isobutyl-1-methylxanthine, 5 μg/mL Insulin, 125 μmol/L indomethacin, 1 μmol/L 3,3′-triiodo-L-thyronine and 0.5 μmol/L Stimulated with a differentiation cocktail containing L rosiglitazone. Cells were maintained with culture medium supplemented with 5 μg/mL insulin, 1 μmol/L 3,3′,5-triiodo-L-thyronine and 1 μmol/L rosiglitazone 2 days after differentiation induction, every 2 days Medium was replaced.

<Preparation Example 2> Diet-induced obesity animal model

In order to confirm the effect of tauroursodeoxycholic acid (TUDCA) of the present invention, a diet-induced obesity animal model (diet-induced obesity, DIO) was prepared. Specifically, 7-week-old male C57BL/6 mice were purchased from Orient (RaonBio, Yongin, South Korea) and fed on a high-fat diet (HFD) with 20% and 60% calories from carbohydrates and fat, respectively. D12492 pellets; Research Diets, New Brunswick, NJ, USA) were supplied. Mice were maintained for 18 weeks on a 12-hour light/dark cycle. In addition, in order to confirm the effect of the pretreatment of TUDCA, NCD (Nomal Control Diet) fed mice (NCD-fed) were used.

In addition, in order to measure the rectal temperature of the mouse, it was measured using a thermometer (Harvard equipment, Edenbridge, Kent, UK) with a rectal probe attached.

<Preparation Example 3> Western blot analysis

In order to confirm the expression of the unfolded protein response (UPR) and the thermogenic programming transcription complex related to the differentiation of beige adipocytes, the expression of related proteins was analyzed by Western blot. Specifically, cell and tissue lysates were prepared using a radioimmunoprecipitation assay (RIPA) buffer containing protease/phosphatase inhibitors, and the total protein concentration of each sample was determined using the Bicinchoninic Acid Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA) was used. Proteins from whole cell and tissue lysates were separated by 10% SDS-PAGE, transferred to a nitrocellulose membrane, and detected using an antibody that specifically binds to the target protein. Then, chemiluminescence was detected using enhanced chemiluminescence, and the nitrocellulose film was exposed to an imaging film and visualized using an automatic film processor (CP1000, Agfa, Mortsel, Belgium). Antibodies for Western blot analysis are shown in Table 1 below.

Antibody Source Application Anti-UCP1 Abcam, ab10983 1:500-10000 Anti-Perilipin Sigma, P1998 1:4000 Anti-VDAC (B-6) Santacruz, sc-390996 1:1000 Anti-p-eIF2α Abcam, ab32157 1:2000 Anti-eIF2α Cell Signaling technology, #9722 1:2000 Anti-KDEL Abcam, ab12223 1:1000 Anti-CHOP Santacruz, sc-575 1:1000 Anti-C/EBPa Santacruz, sc-61 1:500 Anti-HSP90α/β Santacruz, sc-13119 1:3000 Anti-α-TUBULIN Sigma, T9026 1:2000 Anti-Rabbit IgG-HRP Jackson laboratory, 111-035-003 1:2000 Anti-Mouse IgG-HRP Jackson laboratory, 115-035-003 1:2000

<Preparation Example 4> mRNA expression analysis (real time PCR)

In order to confirm the expression of the unfolded protein response (UPR) and the thermogenic programming transcription complex related to the differentiation of beige adipocytes, total RNA expression of related genes was analyzed. Specifically, total RNA was extracted from cells using Trizol (Gene All, Ribo-Ex, Seoul, Korea), and cDNA was extracted using the iScript cDNA Synthesis Kit (BR170-8891, Bio-Rad, Hercules, CA, USA). synthesized. The amount of relative mRNA was calculated as a comparative threshold cycle (Ct) value for 36B4 using the StepOnePlus Real-Time PCR System including SYBR green reagent (RT500M, Enzynomics, Daejeon, Korea). Primer sequences for Real-Time PCR are shown in Table 2 below.

Gene Name Primer sequence Ucp1 5'-ACTGCCACACCTCCAGTCATT -3' 5’- CTTTGCCTCACTCAGGATTGG-3’ Cox8b 5'- TGCTGGAACCATGAAGCCAAC -3' 5'- AGCCAGCCAAAACTCCCACTT -3' Cidea 5'-TCCTATGCTGCACAGATGACG -3' 5'-TGCTCTTCTGTATCGCCCAGT -3' Prdm16 5’-CAGCACGGTGAAGCCATTC-3’ 5’-GCGTGCATCCGCTTGTG-3’ Pgc-1α 5’- AACCACACCCACAGGATCAGA-3’ 5'-TCTTCGCTTTATTGCTCCATGA -3' aP2 5’- AAGGTGAAGAGCATCATAACCCT-3’ 5'- TCACGCCTTTCATAACACATTCC -3' Adiponectin 5'-CTCTAAAGATTGTCAGTGGATCTG -3' 5'-ACGTCATCTTCGGCATGACT -3' Fgf21 5'- CTGCTGGGGGTCTACCAAG -3' 5’-CTGCGCCTACCACTGTTCC-3’ C/ebpα 5’-CCAAGAAGTCGGTGGACAAG-3’ 5’-TTGTTTGGCTTTATCTCGGC-3’ C/ebpβ 5’-GTTTCGGGACTTGATGCAAT-3’ 5’-GGCCCGGCTAGACAGTTAC-3’ C/ebpγ 5'-TTGTAATTGGCCCCAAAGAG -3' 5'-TCAGGGAAATCCAAACTTGC -3' Pparα 5'-GTGGGGAGAGAGGACAGATG-3' 5'-AACTGACGTTTGTGGCTGGT -3' Pparγ 5’-GATGCACTGCCTATGAGCAC-3’ 5’-TCTTCCATCACGGAGAGGTC-3’ Atf3 5’- ATAAACACCTCTGCCATCGG -3’ 5'- GCCTCCTTTTCCTCTCATCTTC -3' Atf4 5'-GAAGAGGTCCGTAAGGCAAG-3' 5’-CAAACACAGCAACACAAGACT-3’ Atf6 5’-CCAACAGAAAGCCCGCATT-3’ 5'-TGGACAGCCATCAGCTGAGA-3' Chop 5’-CTGCCTTTCACCTTGGAGAC-3’ 5'-CGTTTCCTGGGGATGAGATA-3' sXbp1 5'-GAGTCCGCAGCAGGTG-3' 5'-GTGTCAGAGTCCATGGGA-3' tXbp1 5’-AAGAACACGCTTGGGAATGG-3’ 5'- ACTCCCCTTGGCCTCCAC -3' Bip 5’-GGTGCAGCAGGACATCAAGTT-3’ 5’-CCCACCTCCAATATCAACTTGA-3’ 36b4 5'-AACGGCAGCATTTATAACCC-3' 5'-CGATCTGCAGACACACACTG-3'

<Preparation Example 5> Oile red O staining

For differentiation of beige adipocytes and staining of adipocytes, the cells were washed three times with PBS buffer and fixed with formaldehyde for 10 minutes. After that, it was dyed for 10 to 20 minutes using 0.3% Oil red O reagent, and washed three times with distilled water. After that, it was confirmed using a Ti-U microscope system (Nikon, Tokyo, Japan), and a fluorescence value expressed by measuring absorbance at 540 nm using a spectrophotometer (Promega, Madison, WI, USA) for quantification was quantified.

<Preparation Example 6> Preparation of recombinant adenovirus

In order to confirm whether the differentiation of beige adipocytes is affected by the introduction of CHOP, which is known to inhibit the differentiation of beige adipocytes in the unfolded protein response (UPR) related to the differentiation of beige adipocytes, CHOP was A recombinant adenovirus was prepared for introduction into cells. Specifically, in order to prepare an adenovirus expressing green fluorescent protein and CHOP, Murine Chop cDNA was prepared. The Flag-CHOP plasmid was constructed by transferring the prepared Chop cDNA to the pFLAG-CMV-4 expression vector (Sigma). An adenoviral vector expressing CHOP was constructed using AdEasy™ (Qbiogene). Adenoviruses expressing ATF4 and ATF41RK were provided by R. Ratan (Cornell University, USA). EGFP (Enhanced Green Fluorescent protein), a recombinant retroviral vector expressing 299 C-terminal amino acids (299-590) of hamster GADD34 (1N) or 536 N-terminal amino acids of hamster (121 C-terminal amino acid deletion) GADD34 ( 1C) was provided by D. Ron (University of Cambridge, UK). Adenoviruses expressing β-gal and GFP were purchased from the University of Michigan's Viral Vector Core and used.

<Preparation Example 7> Histological analysis and immunofluorescence analysis

Histological analysis was performed using an L710 confocal microscope (Zeiss, Oberkochen, Germany), and lipid accumulation was analyzed using hematoxylin and eosin (H&E) staining of formalin-fixed, paraffin-embedded tissue sections. In addition, for UCP1 analysis, iWAT was obtained from mice, fixed in formalin and embedded in paraffin. Thereafter, the cells were incubated with the primary antibody at 4° C. for 24 hours, reacted with the secondary antibody, and then stained the nuclei with DAPI.

<Preparation Example 8> Confirmation of glucose and insulin resistance

Blood glucose was measured using a Barozen glucometer (Handok, Seoul, Korea) at 0, 30, 60, and 120 after intraperitoneal injection of D-glucose for IPGTT (glucose tolerance test) at a concentration of 2 g/kg. Insulin for the insulin tolerance test (IPITT) was intraperitoneally injected at a concentration of 1 unit/kg and then measured in the same manner.

<Preparation Example 9> Statistical analysis

Statistical analysis was performed using GraphPad Prism 6.0 (GraphPad Inc., San Diego, CA, USA), and all values were expressed as mean ± SEM. Significance between mean values was evaluated by two-tailed unpaired Student's t-test, and it was considered significant when the P value was less than 0.05 (*) and less than 0.01 (**).

<Example 1> Confirmation of suppression of beige adipocyte differentiation by ER-stress

To confirm the role of ER-stress in beige adipogenesis, stromal-vascular fraction cells (SVC) of inguinal white adipose tissue (iWAT) were treated with the chemical ER-stress inducer tunicamycin ( Differentiation of preadipocytes was confirmed according to the presence or absence of Tm) and thapsigargin (Tg) treatment.

In addition, using a recombinant adenoviral vector, beige fat differentiation according to CHOP introduction was confirmed in SVC into which CHOP was introduced.

As a result, as shown in Figure 1A, it was confirmed that tunicamycin (Tm) and thapsigargin (Tg) significantly inhibited beige adipogenesis, and it was confirmed that the expression of UCP1 and C/EBPα was reduced, and UCP1 and C/EBPα were reduced. At the same time as EBPα expression decreased, it was confirmed that the expression of ATF4 and CHOP significantly increased (FIGS. 1B and 1C). In addition, it was confirmed that CHOP introduced into SVC significantly suppressed the expression of beige adipocyte markers UCP1, COX8b, CIDEA, and PGC-1α (FIGS. 1D and 1E). In addition, by the introduced CHOP, PPARα, PPARγ, and C/EBPα, which are generally known to be involved in adipogenesis, were significantly decreased, but there was no significant difference in C/EBPβ and CEBPγ. In addition, the introduced CHOP also significantly suppressed the protein expression of UCP1 and C/EBPa, but not C/EBPβ. From the above results, it was confirmed that ER-stress and the CHOP pathway mediated by ER-stress suppress beige adipocyte differentiation.

<Example 2> Confirmation of beige adipocyte differentiation according to ER-stress reduction

In order to confirm the effect of promoting the differentiation of beige adipocytes of tauroursodeoxycholic acid (TUDCA), a compound known as an ER-stress reducer of the present invention, using SVC in the same manner as in Example 1, ER - Beige adipocyte differentiation according to treatment with a stress reliever was confirmed.

In addition, in order to further confirm the effect of reducing ER-stress, the effect of 4-phenylbutyrate (4-PBA) on promoting differentiation of beige adipocytes was confirmed in the same manner as above.

As a result, as shown in FIG. 2A, in contrast to the ER-stress inducer, in the TUDCA-treated group, the thermogenic programming transcription complex, UCP1, COX8b, CIDEA, PRDM16, PGC-1α and It was confirmed that the expression of C/EBPa was significantly increased. In addition, it was confirmed that the expression of CHOP was significantly decreased (Fig. 2B).

In addition, 4-PBA, which was additionally confirmed, promoted beige fat differentiation similarly to the above TUDCA (Fig. 2C), significantly increased mRNA and protein expression of UCP1, and significantly suppressed CHOP protein expression. It was confirmed (FIGS. 2D and 2E). Based on the above results, it was confirmed that reducing ER-stress activates the differentiation of beige adipocytes.

<Example 3> Confirmation of the effect of reducing ER-stress in the early stage of beige adipocyte differentiation

In order to confirm the relationship between ER-stress/UPR signaling and beige adipogenesis of the present invention, the expression pattern of the UPR gene was examined during the differentiation of beige adipocytes. In addition, in order to confirm whether the TUDCA of the present invention affects the early stage of beige adipocyte differentiation, on day 1 before induction of differentiation (day -1), on the day of differentiation (Day 0), and on day 3 after differentiation (Day 3) TUDCA was treated, respectively, and the expression of markers related to differentiation was confirmed during the differentiation process.

First, the expression pattern of the UPR gene was confirmed in the process of beige adipocyte differentiation. Although the mRNA expressions of ATF4, CHOP, and ATF6 were significantly decreased until the second day of differentiation, it was confirmed that mRNA expression increased from day 6 after differentiation (FIG. 3A). Therefore, it was assumed that the reduction of the UPR gene related to endoplasmic reticulum stress would play an important role in the early stage of the differentiation process of beige adipocytes. On the contrary, it was confirmed that the expression of UCP1 increased from the 6th day after differentiation, and perilipin and VDAC1 increased from the 2nd day of differentiation (Fig. 3B). This means that the differentiation of beige adipocytes was properly performed in the experimental results of the present invention.

Based on these results, it was hypothesized that reducing endoplasmic reticulum stress in the early stage of differentiation of beige adipocytes would promote differentiation, and experiments were conducted to prove this. In the process of beige adipocyte differentiation, TUDCA was treated one day before differentiation (Day -1), simultaneously with differentiation (Day 0), and 3 days after differentiation (Day 3), and differentiation of beige adipocytes was confirmed. As shown in FIG. 3C, when TUDCA was treated on Days -1 and 0, which are the initial stages of differentiation, beige differentiation was significantly promoted, but when treated 3 days after differentiation, it was confirmed that the effect was reduced. Similarly, when TUDCA was treated at the beginning of differentiation (Day -1 and Day 0), it was confirmed that the expression of UPC1 significantly increased, but when treated after 3 days, the effect was confirmed to decrease (Fig. 3D).

Based on the above results, the reduction of UPR and CHOP in the early stage of beige fat differentiation is important in the process of generating beige fat, and as a result, it is important to reduce ER-stress in the early stage of differentiation to inhibit the expression of these genes, resulting in beige It was confirmed that it promotes differentiation of fat.

<Example 4> Confirmation of the effect of TUDCA in high-fat diet mice

<4-1> Confirmation of inhibition of lipid accumulation by administration of TUDCA

The present invention was intended to confirm the browning promotion effect of mouse inguinal white adipose tissue (iWAT) according to the promotion of beige adipocyte differentiation. Specifically, the high-fat diet (high-fat diet, HFD) mice of Preparation Example 2 were used. Specifically, TUDCA was administered to mice fed the HFD diet for an additional 3 weeks after 18 weeks of HFD feeding. Thereafter, the body weight and food intake of the mice were analyzed, and at the end of the experiment, the mice were humanely sacrificed to confirm the effect of TUDCA administration.

As a result, in the mouse group administered with TUDCA, the body weight decreased significantly compared to the control group (FIG. 4A), and there was no significant difference in the change in dietary intake (FIG. 4B). It was confirmed that the body weight was effectively reduced without any change.

In addition, as a result of confirming the tissue weight of iWAT, epididymal white adipose tissue (eWAT), and liver tissue of the sacrificed mouse after the experiment, in the mouse group administered with TUDCA of the present invention, iWAT, eWAT and liver tissue It was confirmed that the weight was significantly reduced (Fig. 4C). In addition, it was confirmed that the fat accumulation in the liver tissue was reduced by the administration of TUDCA (FIG. 4D). Therefore, it was confirmed that the TUDCA of the present invention inhibits lipid accumulation in white tissue and liver tissue following feeding of the high-fat diet.

<4-2> Confirmation of lipid metabolism regulation by administration of TUDCA

It was investigated whether the factors related to glucose absorption, insulin resistance, and fat metabolism were regulated by the administration of TUDCA according to the present invention. Specifically, fasting blood glucose, glucose resistance, and insulin resistance were evaluated using the mice sacrificed in Example 4-1, and it was confirmed whether browning of iWAT was activated according to beige adipogenesis.

As a result, as shown in FIG. 4E, it was confirmed that fasting blood glucose was significantly reduced, and glucose resistance (GTT) and insulin resistance (ITT) were significantly reduced in mice administered with TUDCA of the present invention (FIG. 4F and Figure 4G).

In addition, as a result of confirming the browning of iWAT, it was confirmed that the mRNA and protein expressions of UCP1 and C/EBPα increased significantly in the group administered with TUDCA compared to the control group (FIG. 4H and FIG. 4I), eIF2α, IRE1α , it was confirmed that the protein expression of cleaved ATF6α and CHOP significantly decreased (FIG. 4I). Based on the results of Examples 4-1 and 4-2, it was confirmed that administration of TUDCA of the present invention reduces ER-stress and activates the production of beige adipocytes in mice fed a high-fat diet.

<Example 5> Confirmation of regulation of lipid metabolism according to prior administration of TUDCA

Based on the results of Example 3 and Example 4, it was hypothesized that the reduction of endoplasmic reticulum stress would be more important in the early stages of differentiation or obesity. In other words, compared to HFD after HFD feeding, the administration of TUDCA before feeding improved the browning of iWAT and more effectively suppressed the delay in metabolic disorders caused by HFD. Specifically, TUDCA was administered to mice every day from 3 weeks before feeding the HFD to the end of the experiment, and then, in the same manner as in Example 4, the effect of TUDCA administration before feeding the HFD was confirmed.

As a result, it was confirmed that when TUDCA was administered before HFD feeding, weight gain was significantly inhibited compared to the control group (FIG. 5A). In addition, with prior administration of TUDCA, tissue size and weight, iWAT and epididymal white adipose tissue It was confirmed that white adipose tissue of (epididymal white adipose tissue, eWAT) was reduced (FIG. 5B), and it was confirmed that the size of adipocytes in adipose tissue was reduced (FIG. 5C). In addition, treatment with TUDCA reduced fasting blood sugar. and reduced glucose resistance and insulin resistance (Fig. 5D, Fig. 5E and Fig. 5F).

As a result of confirming the rectal temperature of the mouse, the rectal temperature significantly increased in mice pre-administered with TUDCA (FIG. 5G), and the diameter of iWAT decreased, the density of adipocytes increased, and the number of UCP1-positive cells significantly increased. confirmed (Fig. 5H and Fig. 5I). In addition, the expression of UCP1 and C/EBPa was significantly increased by pre-administration of TUDCA, but it was confirmed that ER-stress and UPR signal transduction pathways CHOP, eIF2α, ATF6α, and IRE1α were significantly decreased (FIG. 5J and FIG. 5K).

Therefore, the present invention confirmed that endoplasmic reticulum stress suppresses the differentiation of beige adipocytes, and first identified that endoplasmic reticulum stress reduces the expression of a thermogenic complex associated with beige adipocyte differentiation. Accordingly, when TUDCA, which reduces endoplasmic reticulum stress, is treated with endoplasmic reticulum stress-induced adipocytes, the expression of factors of the UPR pathway, which is an endoplasmic reticulum stress response, is significantly reduced, and the expression of factors related to UCP1 and the thermogenic complex is increased. Thus, it was confirmed that the differentiation of beige adipocytes was promoted. In addition, in mice treated with TUDCA of the present invention on a high-fat diet, it was confirmed that body weight was reduced, metabolic abnormalities were improved, and beige fat cells in adipose tissue were increased and the size and density of white adipose tissue were decreased. The improvement effect of metabolic diseases induced by stress was confirmed. In particular, it was confirmed that the differentiation of beige fat can be promoted more effectively if TUDCA is pre-treated before the obesity process begins, that is, before ingestion of a high-fat diet. In addition, it was confirmed that the differentiation of beige adipocytes can be more effectively promoted when TUDCA is treated before obesity is generated.

Claims (31)

A pharmaceutical composition for preventing or treating metabolic diseases, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient. According to claim 1,
The metabolic disease is induced by endoplasmic reticulum stress (ER stress), a pharmaceutical composition for preventing or treating metabolic diseases. According to claim 1,
The composition is to increase the formation of beige adipocytes cells, a pharmaceutical composition for preventing or treating metabolic diseases. According to claim 3,
The formation of the beige adipocytes is induced by the expression of a thermogenic transcription complex, a pharmaceutical composition for preventing or treating metabolic diseases. According to claim 1,
The composition is to increase the expression of the thermogenic transcription complex, a pharmaceutical composition for the prevention or treatment of metabolic diseases. According to claim 5,
The thermogenic transcription complex includes uncoupling protein 1 (UCP1), cytochrome c oxidase subunit 8B (COX8b), PR domain containing 16 (PRDM16), Peroxisome proliferator-activated receptor gamma (PPARγ), Peroxisome proliferator-activated receptor gamma alpha (PPARα ), Ccaat-enhancer-binding proteins α (C/EBPα), Ccaat-enhancer-binding proteins β (C/EBPβ), Peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC-1α), Perilipin, Voltage-dependent Anion-selective channel 1 (VDAC1) and a factor selected from the group consisting of Cell Death Inducing DFFA Like Effector A (CIDEA), a pharmaceutical composition for preventing or treating metabolic diseases. According to claim 1,
The composition is to suppress the expression of the endoplasmic reticulum stress factor, a pharmaceutical composition for the prevention or treatment of metabolic diseases. According to claim 7,
The endoplasmic reticulum stress is to induce an unfolded protein response (UPR), a pharmaceutical composition for preventing or treating metabolic diseases. According to claim 7,
The endoplasmic reticulum stress factors are PKR-like ER kinase (PERK), inositol-requiring enzyme 1α (IRE1α), activating transcription factor 6α (ATF6α), activating transcription factor 4 (ATF4), eukaryotic initiation factor 2α (eIF2α) and C/EBP A pharmaceutical composition for the prevention or treatment of metabolic diseases, which is a factor selected from the group consisting of homologous protein (CHOP). According to claim 1,
The composition is to reduce body weight, a pharmaceutical composition for the prevention or treatment of metabolic diseases. According to claim 1,
The composition, to reduce the size or density of fat cells in the tissue, a pharmaceutical composition for the prevention or treatment of metabolic diseases. According to claim 11,
The tissue is inguinal white adipose tissue (iWAT), epididymal white adipose tissue (eWAT) or liver tissue, which is a pharmaceutical composition for preventing or treating metabolic diseases. According to claim 1,
The composition is to inhibit metabolic disorders induced by metabolic diseases, a pharmaceutical composition for the prevention or treatment of metabolic diseases. According to claim 13,
The metabolic abnormality is glucose resistance or insulin resistance is increased, the pharmaceutical composition for the prevention or treatment of metabolic diseases. According to claim 1,
The composition, to reduce fasting blood sugar, a pharmaceutical composition for the prevention or treatment of metabolic diseases. According to claim 1,
The metabolic disease is obesity, type 1 diabetes, type 2 diabetes, hypoglycemia, hypercholesterinemia, hyperlipidemia, hemochromatosis , Amyloidosis (amyloidsis) and porphyria (porphyria) selected from the group consisting of, a pharmaceutical composition for the prevention or treatment of metabolic diseases. A food composition for preventing or improving metabolic disease, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient. A composition for promoting differentiation of beige adipocytes, comprising tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient. According to claim 18,
The composition contains intracellular uncoupling protein 1 (UCP1), Cytochrome c oxidase subunit 8B (COX8b), PR domain containing 16 (PRDM16), Peroxisome proliferator-activated receptor gamma (PPARγ), Peroxisome proliferator-activated receptor gamma alpha (PPARα) , Ccaat-enhancer-binding proteins α (C/EBPα), Ccaat-enhancer-binding proteins β (C/EBPβ), Peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC-1α), Perilipin, Voltage-dependent anion A composition that increases the expression of a factor selected from the group consisting of -selective channel 1 (VDAC1) and Cell Death Inducing DFFA Like Effector A (CIDEA). According to claim 18,
The composition contains intracellular PKR-like ER kinase (PERK), inositol-requiring enzyme 1α (IRE1α), activating transcription factor 6α (ATF6α), activating transcription factor 4 (ATF4), eukaryotic initiation factor 2α (eIF2α) and C/ To reduce the expression of a factor selected from the group consisting of EBP homologous protein (CHOP), the composition. According to claim 18,
The differentiation is dexamethasone, 3-isobutyl-1-methylxanthine, insulin, indomethacin, 3,3'-triiodine-L- A composition derived from a compound selected from the group consisting of tyronine (3,3 ',5-triiodo-L-thyronine) and rosiglitazone. Containing tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt thereof as an active ingredient, the differentiation of beige adipocytes cells is inhibited, and the expression of the thermogenic transcription complex is inhibited. A pharmaceutical composition for the prevention or treatment of metabolic diseases. 23. The method of claim 22,
Wherein the composition increases the expression of a thermogenic transcription complex. 24. The method of claim 23,
The thermogenic transcription complex includes uncoupling protein 1 (UCP1), cytochrome c oxidase subunit 8B (COX8b), PR domain containing 16 (PRDM16), Peroxisome proliferator-activated receptor gamma (PPARγ), Peroxisome proliferator-activated receptor gamma alpha (PPARα ), Ccaat-enhancer-binding proteins α (C/EBPα), Ccaat-enhancer-binding proteins β (C/EBPβ), Peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC-1α), Perilipin, Voltage-dependent A composition that is a factor selected from the group consisting of anion-selective channel 1 (VDAC1) and Cell Death Inducing DFFA Like Effector A (CIDEA). 23. The method of claim 22,
Wherein the composition reduces body weight, the composition. 23. The method of claim 22,
Wherein the composition reduces the size or density of adipocytes in tissues. 27. The method of claim 26,
Wherein the tissue is inguinal white adipose tissue (iWAT), epididymal white adipose tissue (eWAT) or liver tissue. 23. The method of claim 22,
Wherein the composition is to inhibit metabolic disorders induced by metabolic diseases, the composition. 29. The method of claim 28,
The metabolic abnormality is that glucose resistance or insulin resistance is increased, the composition. 23. The method of claim 22,
The composition, to reduce fasting blood sugar, a pharmaceutical composition for the prevention or treatment of metabolic diseases. 23. The method of claim 22,
The metabolic disease is obesity, type 1 diabetes, type 2 diabetes, hypoglycemia, hypercholesterinemia, hyperlipidemia, hemochromatosis , Amyloidosis (amyloidsis) and porphyria (porphyria) selected from the group consisting of, a pharmaceutical composition for the prevention or treatment of metabolic diseases.