WO2022059946A1 - Pharmaceutical composition for treating nonalcoholic steatohepatitis and liver fibrosis - Google Patents

Pharmaceutical composition for treating nonalcoholic steatohepatitis and liver fibrosis Download PDF

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WO2022059946A1
WO2022059946A1 PCT/KR2021/011231 KR2021011231W WO2022059946A1 WO 2022059946 A1 WO2022059946 A1 WO 2022059946A1 KR 2021011231 W KR2021011231 W KR 2021011231W WO 2022059946 A1 WO2022059946 A1 WO 2022059946A1
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pharmaceutical composition
composition
disease
present
nash
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French (fr)
Korean (ko)
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성승용
이송진
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서울대학교 산학협력단
(주)샤페론
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Priority to CA3192690A priority Critical patent/CA3192690A1/en
Priority to KR1020237005249A priority patent/KR20230079001A/en
Publication of WO2022059946A1 publication Critical patent/WO2022059946A1/en

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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 COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • 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
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/30—Other Organic compounds

Definitions

  • the present invention relates to a pharmaceutical composition for the treatment of nonalcoholic steatohepatitis (NASH) and liver fibrosis (Liver Fibrosis), specifically, sodium taurodeoxycholate (Na-TDC), a solvate thereof, or It relates to a pharmaceutical composition comprising a pharmaceutically acceptable salt thereof.
  • NASH nonalcoholic steatohepatitis
  • Liver Fibrosis liver fibrosis
  • Na-TDC sodium taurodeoxycholate
  • a pharmaceutical composition comprising a pharmaceutically acceptable salt thereof.
  • Non-alcoholic fatty liver disease is the most common chronic liver disease and is closely related to obesity, diabetes, and metabolic syndrome. This refers to liver disease in a broad sense, ranging from simple steatosis, in which triglycerides are excessively accumulated in the liver, to cirrhosis of the liver.
  • Nonalcoholic steatohepatitis is a type of nonalcoholic fatty liver disease, and refers to a disease that exhibits pathological phenomena such as expansive degeneration, apoptosis, and inflammatory infiltration, and furthermore, fibrosis. Not all nonalcoholic fatty liver disease progresses to nonalcoholic steatohepatitis, and it takes about 7 years for nonalcoholic fatty liver to progress to nonalcoholic steatohepatitis. This is not easy. In this regard, even in the case of measurement of liver damage indicators such as ALT and AST, and imaging equipment, the accuracy is not high, so the possibility of death from complications is high. In addition, it is reported that 10-29% of patients with nonalcoholic steatohepatitis aggravate cirrhosis within 10 years, and 4-27% of them further progress to liver cancer.
  • 10-2018-0124123 discloses a NASH therapeutic agent comprising obeticholic acid or a pharmaceutically acceptable salt thereof
  • U.S. Patent No. 10457703 discloses a pharmaceutical composition containing a bile acid derivative. It discloses the use of treatment for FXR or TGR5-mediated diseases.
  • An object of the present invention is sodium taurodeoxycholate (Na-TDC) for the prevention and treatment of metabolic diseases, more specifically, for the prevention and treatment of nonalcoholic steatohepatitis (NASH) and liver fibrosis (Liver Fibrosis)
  • Na-TDC sodium taurodeoxycholate
  • NASH nonalcoholic steatohepatitis
  • Liver Fibrosis liver fibrosis
  • the present invention provides a pharmaceutical composition for preventing or treating metabolic diseases comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient do.
  • the metabolic disease is at least one selected from nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver fibrosis (Liver Fibrosis), obesity, diabetes and hyperlipidemia.
  • NAFLD nonalcoholic fatty liver disease
  • NASH nonalcoholic steatohepatitis
  • Liver Fibrosis liver fibrosis
  • obesity diabetes and hyperlipidemia.
  • the metabolic disease is nonalcoholic steatohepatitis (NASH).
  • NASH nonalcoholic steatohepatitis
  • the metabolic disease is Liver Fibrosis.
  • the composition is to reduce steatosis, inflammation or ballooning.
  • the composition reduces fibrosis or cirrhosis.
  • the present invention provides an oral formulation comprising a pharmaceutical composition for preventing or treating the metabolic disease.
  • the oral preparation is selected from the group consisting of tablets, granules, pills, powders, capsules and liquids.
  • the present invention provides an injectable formulation comprising a pharmaceutical composition for the prevention or treatment of the metabolic disease.
  • composition of the present invention may be administered in combination with a therapeutic agent for non-alcoholic fatty liver disease (NAFLD).
  • NAFLD non-alcoholic fatty liver disease
  • composition of the present invention may be administered in combination with a therapeutic agent for non-alcoholic steatohepatitis (NASH).
  • NASH non-alcoholic steatohepatitis
  • the therapeutic agent for nonalcoholic fatty liver disease is OCA (Obeticholic acid), thiazolidinediones, vitamin E, metformin, statin, ursodeoxycholic acid, unsaturated fatty acid, angiotensin receptor blocker , Pentoxifylline, GLP-1 receptor agonists (Glucagon-like peptide 1 receptor agonists), DPP-4 inhibitors (Dipeptidyl peptidase 4 inhibitors), SGLT2 inhibitors (sodium/glucose cotransporter 2 inhibitors), elafibrano (Elafibranor) and telmisartan (Telmisartan) is any one or more selected from the group consisting of.
  • the present invention provides a food composition for preventing or improving metabolic diseases, comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof.
  • the present invention provides a method for treating a metabolic disease comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the present invention can treat non-alcoholic steatohepatitis and liver fibrosis through a pharmaceutical composition comprising sodium taurodeoxycholate (Na-TDC), a solvate thereof, or a pharmaceutically acceptable salt thereof, and have side effects Since there are few, it can be utilized as a pharmaceutical.
  • Na-TDC sodium taurodeoxycholate
  • 1a to 1c are diagrams showing the effect of TDCA in the STAM model.
  • 2a and 2b are diagrams confirming the effect of TDCA through histological analysis of the liver.
  • 3A and 3B are diagrams confirming the fibrosis therapeutic effect of TDCA.
  • 4A and 4B are diagrams confirming the anti-inflammatory effect of TDCA.
  • the present invention relates to a metabolic disease comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient, more specifically, for the prevention or treatment of non-alcoholic steatohepatitis (NASH) It relates to a pharmaceutical composition.
  • NASH non-alcoholic steatohepatitis
  • sodium taurodioxycholate refers to a compound having the following chemical structure.
  • sodium taurodeoxycholate is the same as 2-([3 ⁇ ,12 ⁇ -Dihydroxy-24-oxo-5 ⁇ -cholan-24-yl]amino)ethanesulfonic acid, Taurodeoxycholic acid sodium salt hydrate, CAS number 207737-97-1 can be named
  • sodium taurodioxycholate may exist in all forms, for example, it may exist in the form of a crystalline form, an amorphous form, a solvate, etc.
  • solvate refers to a form formed by adding a solvent to a compound, and includes monohydrate, dihydrate, and the like.
  • pharmaceutically acceptable means that the included ingredients do not significantly stimulate the organism and do not impair biological activity and properties.
  • the term “pharmaceutically acceptable salt” refers to a salt having desirable biological activity, but is not limited thereto, but is not limited thereto, but includes inorganic acid salts (hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid), organic acid salts (acetic acid, oxalic acid, maleic acid, fumaric acid, Succinic acid, benzoic acid, ascorbic acid, tannic acid, pamoic acid, alginic acid, triethylamine, cyclohexylamine, pyridine), alkali metal salts (sodium salt, potassium salt), alkaline earth metal salt (calcium salt), ammonium salt or addition salts thereof etc.
  • inorganic acid salts hydroochloric acid, sulfuric acid, phosphoric acid, nitric acid
  • organic acid salts acetic acid, oxalic acid, maleic acid, fumaric acid, Succinic acid, benzoic acid, ascorbic acid, tannic acid
  • prevention refers to any action of suppressing the symptoms of a specific disease (eg, non-alcoholic steatohepatitis) or delaying the progression by injecting the composition of the present invention into the body.
  • a specific disease eg, non-alcoholic steatohepatitis
  • treatment refers to any action that improves or beneficially changes the symptoms of a specific disease (eg, non-alcoholic steatohepatitis) by injecting the composition of the present invention into the body.
  • a specific disease eg, non-alcoholic steatohepatitis
  • the present invention relates to a pharmaceutical composition for preventing or treating metabolic diseases comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the metabolic disease refers to any disease that occurs due to an abnormality in the pathway of a chemical reaction occurring in the body, and includes the following diseases as non-limiting examples of the metabolic disease.
  • metabolic diseases include, for example, type 1 diabetes, type 2 diabetes, hyperlipidemia, hypertension, insulin resistance, obesity, glucose metabolism abnormality, diabetic retinopathy, diabetic nephritis, diabetic neuropathy, body mass gain, Hypercholesterolemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver fibrosis (Liver Fibrosis), polycystic ovary syndrome (PCOS), liver cirrhosis, hepatitis C, alcoholic liver disease, primary sclerosing cholangitis, primary biliary cholangitis; and the like.
  • NAFLD nonalcoholic fatty liver disease
  • NASH nonalcoholic steatohepatitis
  • Liver Fibrosis liver fibrosis
  • PCOS polycystic
  • the pharmaceutical composition of the present invention prevents at least one selected from the group consisting of nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver fibrosis, obesity, diabetes and hyperlipidemia. or a therapeutic effect.
  • NAFLD nonalcoholic fatty liver disease
  • NASH nonalcoholic steatohepatitis
  • liver fibrosis obesity, diabetes and hyperlipidemia. or a therapeutic effect.
  • the pharmaceutical composition of the present invention exhibits a preventive or therapeutic effect of nonalcoholic steatohepatitis (NASH).
  • NASH nonalcoholic steatohepatitis
  • the pharmaceutical composition of the present invention exhibits a preventive or therapeutic effect on liver fibrosis (Liver Fibrosis).
  • the composition is to reduce steatosis, inflammation or ballooning.
  • the composition is to reduce fibrosis or cirrhosis.
  • the pharmaceutical composition of the present invention can be administered orally or parenterally according to a desired method, and the dosage depends on the patient's weight, age, health condition, diet, administration time, administration method, disease type and severity of disease, etc. It can be administered by varying the range.
  • the daily dose is about 0.01 to 1000 mg/kg, and more specifically, 0.1 to 100 mg/kg, and may be administered once to several times a day.
  • an oral preparation comprising a pharmaceutical composition for preventing or treating the metabolic disease.
  • the oral preparation is selected from the group consisting of tablets, granules, pills, powders, capsules and liquids.
  • the present invention provides an injectable formulation comprising a pharmaceutical composition for the prevention or treatment of metabolic diseases.
  • the pharmaceutical preparation for oral administration may be in the form of dosage units such as tablets, granules, pills, powders, capsules or liquids, and ampoules. They are prepared by methods known per se, for example by customary mixing, granulation, confection, dissolution or lyophilization methods.
  • Other orally administrable pharmaceutical preparations include capsules, dry-filled capsules made of gelatin, and soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol.
  • Dry-filled capsules contain the active ingredient in the form of particles, for example in mixture with fillers such as lactose, binders such as starches, and/or glidants such as talc or magnesium stearate, and if appropriate stabilizers You may.
  • the active ingredient is dissolved or suspended in a suitable liquid, such as fatty oil, paraffin oil or liquid polyethylene glycol, to which a stabilizing agent may also be added.
  • a solvent, solubilizer or emulsifier is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- butyl glycol oil, glycerol fatty ester, polyethylene glycol or fatty acid ester of sorbitan.
  • parenteral administration may be, but is not limited to, for example, intravenous, subcutaneous, intradermal, intraperitoneal or topical application.
  • Pharmaceutical preparations for parenteral administration may be sterile and/or contain additives such as preservatives, stabilizers, wetting and/or emulsifying agents, solubilizing agents, salts for adjusting the osmotic pressure and/or buffers.
  • composition of the present invention may be administered in combination with a therapeutic agent for non-alcoholic fatty liver disease (NAFLD).
  • NAFLD non-alcoholic fatty liver disease
  • composition of the present invention may be administered in combination with a therapeutic agent for non-alcoholic steatohepatitis (NASH).
  • NASH non-alcoholic steatohepatitis
  • the therapeutic agent for nonalcoholic fatty liver disease is OCA (Obeticholic acid), thiazolidinediones, vitamin E, metformin, statin, ursodeoxycholic acid, unsaturated fatty acid, angiotensin receptor blocker , Pentoxifylline, GLP-1 receptor agonists (Glucagon-like peptide 1 receptor agonists), DPP-4 inhibitors (Dipeptidyl peptidase 4 inhibitors), SGLT2 inhibitors (sodium/glucose cotransporter 2 inhibitors), elafibrano (Elafibranor) and telmisartan (Telmisartan) is any one or more selected from the group consisting of.
  • the present invention relates to a food composition for preventing or improving metabolic diseases, comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof.
  • the present invention provides a method for treating a metabolic disease comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  • Sodium Taurodeoxycholate is manufactured by New Zealand Pharmaceuticals Ltd. (Palmerston North, New Zealand). Telmisartan (Micardis®) was purchased from Boehringer Ingelheim GmbH (Germany) and dissolved in purified water. DPBS (Dulbecco's Phosphate-Buffered Salines) was purchased from WelGene Inc., (LB001-02, Korea) and used as a vehicle compound.
  • the STAM model was induced in C57BL/6 mice through a combination of a high-fat diet and the chemical streptozotocin.
  • C57BL/6 mice 14 days pregnant female
  • SMC Laboratories, Inc. Tokyo, Japan
  • All animals used in the study were housed and managed according to the Animal Use Guidelines of the Japanese Pharmacological Association.
  • nonalcoholic steatohepatitis was treated with a single subcutaneous injection of 200 ⁇ g streptozotocin (STZ, Sigma-Aldrich, USA) solution 2 days after birth, and a high-fat diet (HFD, 57 kcal%fat, CLEA) after 4 weeks of age.
  • HFD high-fat diet
  • Japan, Cat# HFD32, Japan was induced free of charge. Mice were randomly assigned, and the specific study design is shown in Figure 1a.
  • vehicle compounds DPBS, TDCA (Taurodeoxycholic acid), and telmisartan were orally administered daily, and treatment was performed until 9 weeks of age. In addition, body weight was measured before each treatment. Thereafter, 9-week-old mice were euthanized by exsanguination by direct cardiac puncture under isoflurane (Pfizer Inc., USA) anesthesia.
  • NAFLD activity score (NAFLD activity score, NAS) was calculated according to the Kleiner criteria.
  • fibrosis area For quantitative analysis of the fibrosis area, a section stained with Sirius Red was captured around the central vein using a digital camera (DFC295; Leica, Germany) at 200x magnification. In addition, positive areas in 5 fields/section were measured using Image J software (National Institute of Health, USA).
  • the fragment was incubated with a biotin-conjugated secondary antibody (VECTASTAIN Elite ABC kit, Vector Laboratories, Cat#: PK-4004, USA), and then incubated with ABC reagent at room temperature for 30 minutes. cultured.
  • a biotin-conjugated secondary antibody VECTASTAIN Elite ABC kit, Vector Laboratories, Cat#: PK-4004, USA
  • the enzyme substrate reaction was performed using a 3,3′-diaminobenzidine/H 2 O 2 solution (3,3′-diaminobenzidine/H 2 O 2 solution) (Nichirei Bioscience inc., Japan).
  • ER-TR7 positive regions For quantitative analysis of ER-TR7 positive regions, bright field images of ER-TR7 immunostained sections were captured around the central vein using a 200x digital camera (DFC295; Leica). In addition, positive areas in 5 fields/section were measured using Image J software (National Institute of Health, USA).
  • RNA expression 60S acidic ribosomal protein P0 (RPLP0) RNA expression was measured as an internal control.
  • Relative expression levels of target genes were compared after normalization to RPLP0. Relative quantification was performed using the ⁇ Ct method.
  • the primer sequence is as follows: CCL2(MCP1) (Forward: 5'-AAACTGCATCTGCCCTAAGG-3' / Reverse: 5'-AGAAGTGCTTGAGGTGGTTG-3').
  • TNF- ⁇ in the mouse plasma was analyzed using the mouse TNF- ⁇ uncoated ELISA Kit (Invitrogen, Cat# 88-7324, USA), and TIMP1 was analyzed through the mouse TIMP1 ELSIA Kit (abcam, Cat# ab196265, Cambridge, UK), The kit was used according to the manufacturer's instructions.
  • TDCA The effect of TDCA was investigated in the STAM model of nonalcoholic steatohepatitis by the experimental method according to ⁇ Experimental Example 1>.
  • Telmisartan an angiotensin II receptor antagonist and peroxisome proliferator-activated receptor- ⁇ agonist, reduces fibrosis and inflammation It has the characteristic of improving NASH.
  • telmisartan was used as a reference drug.
  • TDCA liver to body weight ratio
  • DPBS vehicle group
  • the STAM model group had hepatocyte steatosis, ballooning, and inflammatory cell infiltration (FIG. 2A).
  • the TDCA 1.25 mg/kg treatment group and the telmisartan group showed a significant decrease in total NAS, confirming the improvement of the NAS lesion index, an important clinical symptom measure of NASH ( FIG. 2B ).
  • liver sections stained with Sirius Red and anti-fibroblast antibody ER-TR7 stained liver sections from STAM mice showed increased collagen deposition in the hepatic lobule region compared to normal mice (Fig. 3A).
  • the mRNA expression level of mouse liver tissue genes related to inflammation and fibrosis in NASH was analyzed according to the above experimental example.
  • the analyzed inflammatory marker is monocyte chemoattractant protein-1 (MCP-1).
  • MCP-1 monocyte chemoattractant protein-1
  • TDCA 2.5 mg/kg treatment group showed a significant decrease in TNF- ⁇ compared to the vehicle group (DPBS), confirming that there was an anti-inflammatory effect of suppressing important inflammatory cytokines (FIG. 4B).
  • the present invention can be usefully used for the prevention or treatment of metabolic diseases, more specifically, the prevention or treatment of nonalcoholic steatohepatitis (NASH).
  • NASH nonalcoholic steatohepatitis

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Abstract

The present invention relates to a pharmaceutical composition for preventing and treating metabolic diseases, and more specifically, for treating nonalcoholic steatohepatitis (NASH) or liver fibrosis through the pharmaceutical composition comprising sodium taurodeoxycholate (Na-TDC), a solvate thereof or a pharmaceutically acceptable salt thereof.

Description

비알코올성 지방간염 및 간 섬유화의 치료를 위한 약학적 조성물Pharmaceutical composition for the treatment of nonalcoholic steatohepatitis and liver fibrosis
본 발명은 비알코올성 지방간염(NASH) 및 간 섬유화(Liver Fibrosis)의 치료를 위한 약학적 조성물에 관한 것으로서, 상세하게는 소듐 타우로디옥시콜레이트(Sodium Taurodeoxycholate, Na-TDC), 이의 용매화물, 또는 이의 약학적으로 허용가능한 염을 포함하는 약학적 조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for the treatment of nonalcoholic steatohepatitis (NASH) and liver fibrosis (Liver Fibrosis), specifically, sodium taurodeoxycholate (Na-TDC), a solvate thereof, or It relates to a pharmaceutical composition comprising a pharmaceutically acceptable salt thereof.
비알코올성 지방간질환(Non-alcoholic fatty liver disease, NAFLD)은 만성 간 질환 중 가장 흔한 질환으로, 비만, 당뇨병, 대사증후군과 밀접한 관계가 있는 질환에 해당한다. 이는 간 내에서 중성 지방이 과다하게 축적되어 있는 단순 지방증(simple steatosis)에서 질환의 심화된 간경변증(Cirrhosis)까지를 아우르는 광의의 간질환을 의미한다.Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and is closely related to obesity, diabetes, and metabolic syndrome. This refers to liver disease in a broad sense, ranging from simple steatosis, in which triglycerides are excessively accumulated in the liver, to cirrhosis of the liver.
비알코올성 지방간염(Nonalocoholic steatohepatitis, NASH)은 비알코올성 지방간질환의 일종으로, 팽창성 변성, 세포사멸, 염증성 침윤 등의 병리학적 현상을 나타내며, 더 나아가 섬유화 현상까지 나타내는 질환을 의미한다. 모든 비알코올성 지방간질환이 비알코올성 지방간염으로 진행되는 것은 아니고, 비알코올성 지방간이 비알코올성 지방간염으로 진행되는데 약 7년 정도가 걸리며, 비알코올성 지방간염 환자의 경우에도 50% 이상은 무증상으로 나타나 진단이 쉽지 않다. 이와 관련하여 ALT, AST와 같은 간손상지표, 영상장비 등의 측정의 경우에도 정확성이 높지 않아 합병증으로 사망할 가능성이 높다. 또한, 비알코올성 지방간염 환자의 10~29%가 10년 이내 간경변으로 심화되고, 그 중 4~27%는 더 나아가 간암으로 진행되는 것으로 보고되고 있다.Nonalcoholic steatohepatitis (NASH) is a type of nonalcoholic fatty liver disease, and refers to a disease that exhibits pathological phenomena such as expansive degeneration, apoptosis, and inflammatory infiltration, and furthermore, fibrosis. Not all nonalcoholic fatty liver disease progresses to nonalcoholic steatohepatitis, and it takes about 7 years for nonalcoholic fatty liver to progress to nonalcoholic steatohepatitis. This is not easy. In this regard, even in the case of measurement of liver damage indicators such as ALT and AST, and imaging equipment, the accuracy is not high, so the possibility of death from complications is high. In addition, it is reported that 10-29% of patients with nonalcoholic steatohepatitis aggravate cirrhosis within 10 years, and 4-27% of them further progress to liver cancer.
비알코올성 지방간염의 원인이 직접적으로 알려진 것은 없으나, 산화스트레스, 지방과산화, 싸이토카인의 손상, 지질대사이상, 인슐린저항성 등의 대사불균형과 관련이 있는 것으로 추측되고 있으며, 이를 치료하기 위한 노력이 국내외에서 다방면으로 이루어지고 있다. 미국의 경우 최근 FDA에서 NASH 신약개발 촉진 지침을 내놓았고, 다국적 제약사인 인터셉트파마슈티컬즈, 길리어드, 엘러간, NGM 바이오파마슈티컬즈 등이 임상시험을 통해 비알코올성 지방간염을 치료하려는 노력이 이루어지고 있다. 이와 관련하여, 지질대사 이상을 개선하는 약제로서, PPARα를 활성화하는 피브레이트류, PPARγ 글리타존류가 알려진바 있다. 또한, 대한민국공개특허 제10-2018-0124123호에서는 오베티콜산 또는 이의 약학적으로 허용 가능한 염을 포함하는 NASH 치료제를 개시하고 있으며, 미국등록특허 제10457703호에서는 담즙산 유도체를 포함하는 약학조성물을 통해 통하여 FXR 또는 TGR5-매개 질환의 치료용도를 개시하고 있다.Although the cause of nonalcoholic steatohepatitis is not known directly, it is speculated that it is related to metabolic imbalance such as oxidative stress, fat peroxidation, cytokine damage, lipid metabolism abnormality, and insulin resistance. It is done in many ways. In the US, the FDA recently issued guidelines for promoting NASH drug development, and multinational pharmaceutical companies such as Intercept Pharmaceuticals, Gilead, Allergan, and NGM Biopharmaceuticals made efforts to treat nonalcoholic steatohepatitis through clinical trials. is losing In this regard, fibrates that activate PPARα and PPARγ glitazones have been known as drugs for improving lipid metabolism abnormalities. In addition, Korean Patent Application Laid-Open No. 10-2018-0124123 discloses a NASH therapeutic agent comprising obeticholic acid or a pharmaceutically acceptable salt thereof, and U.S. Patent No. 10457703 discloses a pharmaceutical composition containing a bile acid derivative. It discloses the use of treatment for FXR or TGR5-mediated diseases.
다만, 다양한 물질의 비알코올성 지방간질환의 치료용도가 알려져 있더라도, 현재까지 유의미한 효과를 나타내는 치료제가 개발되지 않아 치료제에 대한 개발이 절실한 상황이다. 또한, 소듐 타우로디옥시콜레이트를 이용하여 비알코올성 지방간질환의 치료용도도 알려진 바가 없다. 이에, 본 발명자들은 소듐 타우로디옥시콜레이트를 통해 비알코올성 지방간질환 및 간 섬유화의 치료 용도를 밝히기 위해 노력한 결과, 소듐 타우로디옥시콜레이트, 이의 용매화물, 또는 이의 약학적으로 허용가능한 염을 포함하는 약학적 조성물을 치료 용도를 밝혀냄으로써 본 발명을 완성하였다.However, even though various substances are known for the treatment of nonalcoholic fatty liver disease, there has been no development of a therapeutic agent showing a significant effect so far, and the development of a therapeutic agent is urgently needed. In addition, the use of sodium taurodioxycholate for the treatment of non-alcoholic fatty liver disease is not known. Accordingly, as a result of the present inventors' efforts to reveal the therapeutic use of sodium taurodioxycholate for nonalcoholic fatty liver disease and liver fibrosis, sodium taurodioxycholate, a solvate thereof, or a pharmaceutical containing a pharmaceutically acceptable salt thereof The present invention has been accomplished by discovering a therapeutic use of an enemy composition.
본 발명의 목적은 대사성 질환의 예방 및 치료, 보다 상세하게는 비알코올성 지방간염(NASH) 및 간 섬유화(Liver Fibrosis)의 예방 및 치료를 위한 소듐 타우로디옥시콜레이트(Sodium Taurodeoxycholate, Na-TDC), 이의 용매화물 또는 이의 약학적으로 허용가능한 염을 포함하는 약학적 조성물을 제공하는 것이다.An object of the present invention is sodium taurodeoxycholate (Na-TDC) for the prevention and treatment of metabolic diseases, more specifically, for the prevention and treatment of nonalcoholic steatohepatitis (NASH) and liver fibrosis (Liver Fibrosis) To provide a pharmaceutical composition comprising a solvate thereof or a pharmaceutically acceptable salt thereof.
상기 목적을 달성하기 위하여, 본 발명은 소듐 타우로디옥시콜레이트(sodium taurodeoxycholate), 이의 용매화물, 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 질환의 예방 또는 치료용 약학적 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating metabolic diseases comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient do.
본 발명의 일 양태에서, 상기 대사성 질환은 비알코올성 지방간 질병(NAFLD), 비알코올성 지방간염(NASH), 간 섬유화(Liver Fibrosis), 비만, 당뇨병 및 고지혈증으로부터 선택된 1종 이상이다.In one embodiment of the present invention, the metabolic disease is at least one selected from nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver fibrosis (Liver Fibrosis), obesity, diabetes and hyperlipidemia.
본 발명의 일 양태에서, 상기 대사성 질환은 비알코올성 지방간염(NASH)이다.In one embodiment of the present invention, the metabolic disease is nonalcoholic steatohepatitis (NASH).
본 발명의 일 양태에서, 상기 대사성 질환은 간 섬유화(Liver Fibrosis)이다.In one aspect of the present invention, the metabolic disease is Liver Fibrosis.
본 발명의 일 양태에서, 상기 조성물은 지방증(steatosis), 염증(inflammation) 또는 벌룬 현상(ballooning)을 감소시키는 것이다.In one aspect of the present invention, the composition is to reduce steatosis, inflammation or ballooning.
본 발명의 일 양태에서, 상기 조성물은 섬유화(fibrosis) 또는 경화(cirrhosis)를 감소시키는 것이다.In one aspect of the present invention, the composition reduces fibrosis or cirrhosis.
또한, 본 발명은 상기 대사성 질환의 예방 또는 치료용 약학적 조성물을 포함하는 경구용 제제를 제공한다. 구체적인 본 발명의 일 양태에서, 상기 경구용 제제는 정제, 과립제, 환제, 산제, 캅셀제 및 액제로 구성된 그룹으로부터 선택되는 것이다.In addition, the present invention provides an oral formulation comprising a pharmaceutical composition for preventing or treating the metabolic disease. In a specific embodiment of the present invention, the oral preparation is selected from the group consisting of tablets, granules, pills, powders, capsules and liquids.
또한, 본 발명은 상기 대사성 질환의 예방 또는 치료용 약학적 조성물을 포함하는 주사용 제제를 제공한다.In addition, the present invention provides an injectable formulation comprising a pharmaceutical composition for the prevention or treatment of the metabolic disease.
또한, 본 발명 조성물은 비알코올성 지방간 질환(non-alcoholic fatty liver disease; NAFLD) 치료제와 병용 투여될 수 있다. 또한, 본 발명 조성물은 비알코올성 지방간염(non-alcoholic steatohepatitis; NASH) 치료제와 병용 투여될 수 있다.In addition, the composition of the present invention may be administered in combination with a therapeutic agent for non-alcoholic fatty liver disease (NAFLD). In addition, the composition of the present invention may be administered in combination with a therapeutic agent for non-alcoholic steatohepatitis (NASH).
본 발명의 일 양태에서, 상기 비알코올성 지방간 질환 치료제는 OCA (Obeticholic acid), 티아졸리딘디온(Thiazolidinediones), 비타민 E, 메트포르민, 스타틴, 우르소디옥시콜산(Ursodeoxycholic acid), 불포화 지방산, 안지오텐신 수용체 차단제, 펜톡시필린(Pentoxifylline), GLP-1 수용체 아고니스트(Glucagon-like peptide 1 receptor agonists), DPP-4 억제제(Dipeptidyl peptidase 4 inhibitors), SGLT2 억제제(sodium/glucose cotransporter 2 inhibitors), 엘라피브라노(Elafibranor) 및 텔미사르탄(Telmisartan)으로 이루어지는 군으로부터 선택되는 어느 하나 이상이다.In one embodiment of the present invention, the therapeutic agent for nonalcoholic fatty liver disease is OCA (Obeticholic acid), thiazolidinediones, vitamin E, metformin, statin, ursodeoxycholic acid, unsaturated fatty acid, angiotensin receptor blocker , Pentoxifylline, GLP-1 receptor agonists (Glucagon-like peptide 1 receptor agonists), DPP-4 inhibitors (Dipeptidyl peptidase 4 inhibitors), SGLT2 inhibitors (sodium/glucose cotransporter 2 inhibitors), elafibrano (Elafibranor) and telmisartan (Telmisartan) is any one or more selected from the group consisting of.
또한, 본 발명은 소듐 타우로디옥시콜레이트(sodium taurodeoxycholate), 이의 용매화물, 또는 이의 식품학적으로 허용가능한 염을 포함하는, 대사성 질환의 예방 또는 개선용 식품 조성물을 제공한다.In addition, the present invention provides a food composition for preventing or improving metabolic diseases, comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof.
또한, 본 발명은 소듐 타우로디옥시콜레이트(sodium taurodeoxycholate), 이의 용매화물, 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 질환의 치료 방법을 제공한다.In addition, the present invention provides a method for treating a metabolic disease comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명은 소듐 타우로디옥시콜레이트(Sodium Taurodeoxycholate, Na-TDC), 이의 용매화물 또는 이의 약학적으로 허용가능한 염을 포함하는 약학적 조성물을 통해 비알코올성 지방간염 및 간 섬유화를 치료할 수 있고, 부작용이 거의 없으므로, 의약품으로서 활용될 수 있다.The present invention can treat non-alcoholic steatohepatitis and liver fibrosis through a pharmaceutical composition comprising sodium taurodeoxycholate (Na-TDC), a solvate thereof, or a pharmaceutically acceptable salt thereof, and have side effects Since there are few, it can be utilized as a pharmaceutical.
도 1a 내지 도 1c STAM 모델에서 TDCA의 효과를 나타낸 도이다.1a to 1c are diagrams showing the effect of TDCA in the STAM model.
도 2a 및 도 2b는 간의 조직학적 분석을 통해 TDCA의 효과를 확인한 도이다.2a and 2b are diagrams confirming the effect of TDCA through histological analysis of the liver.
도 3a 및 도 3b는 TDCA의 섬유증 치료효과를 확인한 도이다.3A and 3B are diagrams confirming the fibrosis therapeutic effect of TDCA.
도 4a 및 도 4b는 TDCA의 항염증 효과를 확인한 도이다.4A and 4B are diagrams confirming the anti-inflammatory effect of TDCA.
본 발명은 소듐 타우로디옥시콜레이트(sodium taurodeoxycholate), 이의 용매화물, 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 질환, 보다 구체적으로는 비알코올성 지방간염(NASH)의 예방 또는 치료용 약학적 조성물에 관한 것이다.The present invention relates to a metabolic disease comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient, more specifically, for the prevention or treatment of non-alcoholic steatohepatitis (NASH) It relates to a pharmaceutical composition.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명의 명세서 전체에서, 어떤 부분이 어떤 구성 요소를 “포함”한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성 요소를 제외하는 것이 아니라 다른 구성 요소를 더 포함할 수 있는 것을 의미한다. Throughout the specification of the present invention, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.
본 발명에서 사용되는 “소듐 타우로디옥시콜레이트”는 하기 화학 구조를 가진 화합물을 의미한다.As used herein, “sodium taurodioxycholate” refers to a compound having the following chemical structure.
Figure PCTKR2021011231-appb-img-000001
Figure PCTKR2021011231-appb-img-000001
또한, 소듐 타우로디옥시콜레이트는 2-([3α,12α-Dihydroxy-24-oxo-5β-cholan-24-yl]amino)ethanesulfonic acid, Taurodeoxycholic acid sodium salt hydrate, CAS 넘버 207737-97-1와 동일하게 명명될 수 있다.In addition, sodium taurodeoxycholate is the same as 2-([3α,12α-Dihydroxy-24-oxo-5β-cholan-24-yl]amino)ethanesulfonic acid, Taurodeoxycholic acid sodium salt hydrate, CAS number 207737-97-1 can be named
본 발명에서 소듐 타우로디옥시콜레이트는 모든 형태로 존재할 수 있고, 예를 들면 결정형, 비결정형, 용매화물 등의 형태로 존재할 수 있다.In the present invention, sodium taurodioxycholate may exist in all forms, for example, it may exist in the form of a crystalline form, an amorphous form, a solvate, etc.
본 명세서에서 “용매화물”이란 화합물에 용매를 부가하여 형성되는 형태로서, 일수화물, 이수화물 등을 포함한다.As used herein, the term “solvate” refers to a form formed by adding a solvent to a compound, and includes monohydrate, dihydrate, and the like.
본 명세서에서 “약학적으로 허용가능한”이란 포함된 성분이 생물체를 상당히 자극하지 않고, 생물학적 활성 및 특성을 저해하지 않는 것을 의미한다.As used herein, "pharmaceutically acceptable" means that the included ingredients do not significantly stimulate the organism and do not impair biological activity and properties.
본 명세서에서 “약학적으로 허용가능한 염”이란 바람직한 생물활성을 보유한 염으로서, 이에 한정되는 것은 아니나, 무기산염(염산, 황산, 인산, 질산), 유기산염(아세트산, 옥살산, 말레산, 푸마르산, 숙신산, 벤조산, 아스코르브산, 타닌산, 파모산, 알긴산, 트리에틸아민, 사이클로헥실아민, 피리딘), 알칼리금속염(나트륨염, 칼륨염), 알칼리토금속염(칼슘염), 암모늄염 또는 이들의 부가 염 형태 등을 포함한다.As used herein, the term “pharmaceutically acceptable salt” refers to a salt having desirable biological activity, but is not limited thereto, but is not limited thereto, but includes inorganic acid salts (hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid), organic acid salts (acetic acid, oxalic acid, maleic acid, fumaric acid, Succinic acid, benzoic acid, ascorbic acid, tannic acid, pamoic acid, alginic acid, triethylamine, cyclohexylamine, pyridine), alkali metal salts (sodium salt, potassium salt), alkaline earth metal salt (calcium salt), ammonium salt or addition salts thereof etc.
본 명세서에서 “예방”은 본 발명의 조성물의 체내 투입으로 특정 질환(예를 들어, 비알코올성 지방간염)의 증상을 억제시키거나 진행을 지연시키는 모든 행위를 의미한다.As used herein, “prevention” refers to any action of suppressing the symptoms of a specific disease (eg, non-alcoholic steatohepatitis) or delaying the progression by injecting the composition of the present invention into the body.
본 명세서에서 “치료”는 본 발명의 조성물의 체내 투입으로 특정 질환(예를 들어, 비알코올성 지방간염)의 증상을 호전 또는 이롭게 변경시키는 모든 행위를 의미한다.As used herein, “treatment” refers to any action that improves or beneficially changes the symptoms of a specific disease (eg, non-alcoholic steatohepatitis) by injecting the composition of the present invention into the body.
본 명세서에서 기재되는 화합물은 특별히 정의를 하지 않는 한, 통상적으로 인식된 화합물을 의미한다.Compounds described herein refer to commonly recognized compounds unless otherwise defined.
본 발명은 소듐 타우로디옥시콜레이트(sodium taurodeoxycholate), 이의 용매화물, 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 질환의 예방 또는 치료용 약학적 조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for preventing or treating metabolic diseases comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명에서 대사성 질환은 체내에서 일어나는 화학반응의 경로에 이상이 생겨 발생하는 모든 질환을 의미하며, 대사성 질환의 비제한적인 예시에 하기의 질환들을 포함한다. 이에 제한되는 것은 아니나 대사성 질환은 예를 들면, 제1형 당뇨병, 제2형 당뇨병, 고지혈증, 고혈압, 인슐린 저항성, 비만, 당대사 이상, 당뇨병성 망막증, 당뇨병성 신장염, 당뇨병성 신경증, 체질량증가, 고콜레스테롤혈증, 비알코올성 지방간 질환(NAFLD), 비알코올성 지방간염(NASH), 간 섬유화(Liver Fibrosis), 다낭성난소증후군(PCOS), 간 경변증, C형 간염, 알코올성 간 질환, 원발성 경화성 담관염, 원발성 담즙성 담관염 등을 포함한다.In the present invention, the metabolic disease refers to any disease that occurs due to an abnormality in the pathway of a chemical reaction occurring in the body, and includes the following diseases as non-limiting examples of the metabolic disease. Although not limited thereto, metabolic diseases include, for example, type 1 diabetes, type 2 diabetes, hyperlipidemia, hypertension, insulin resistance, obesity, glucose metabolism abnormality, diabetic retinopathy, diabetic nephritis, diabetic neuropathy, body mass gain, Hypercholesterolemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver fibrosis (Liver Fibrosis), polycystic ovary syndrome (PCOS), liver cirrhosis, hepatitis C, alcoholic liver disease, primary sclerosing cholangitis, primary biliary cholangitis; and the like.
본 발명의 일 양태에서, 본 발명 약학적 조성물은 비알코올성 지방간 질병(NAFLD), 비알코올성 지방간염(NASH), 간 섬유화(Liver Fibrosis), 비만, 당뇨병 및 고지혈증으로 구성된 그룹으로부터 선택된 1종 이상의 예방 또는 치료효과를 나타낸다.In one aspect of the present invention, the pharmaceutical composition of the present invention prevents at least one selected from the group consisting of nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver fibrosis, obesity, diabetes and hyperlipidemia. or a therapeutic effect.
구체적인 본 발명의 일 양태에서, 본 발명 약학적 조성물은 비알코올성 지방간염(NASH)의 예방 또는 치료효과를 나타낸다.In a specific aspect of the present invention, the pharmaceutical composition of the present invention exhibits a preventive or therapeutic effect of nonalcoholic steatohepatitis (NASH).
구체적인 본 발명의 일 양태에서, 본 발명 약학적 조성물은 간 섬유화(Liver Fibrosis)의 예방 또는 치료효과를 나타낸다.In a specific aspect of the present invention, the pharmaceutical composition of the present invention exhibits a preventive or therapeutic effect on liver fibrosis (Liver Fibrosis).
본 발명의 일 양태에서, 상기 조성물은 지방증(steatosis), 염증(inflammation) 또는 벌룬 현상(ballooning)을 감소시키는 것이다.In one aspect of the present invention, the composition is to reduce steatosis, inflammation or ballooning.
또한, 본 발명의 일 양태에서, 상기 조성물은 섬유화(fibrosis) 또는 경화(cirrhosis)를 감소시키는 것이다.Also, in one aspect of the present invention, the composition is to reduce fibrosis or cirrhosis.
본 발명의 약학적 조성물은 목적하는 방법에 따라 경구 투여 또는 비경구 투여 할 수 있으며, 투여량은 환자의 체중, 연령, 건강상태, 식이, 투여시간, 투여방법, 질환종류 및 질환의 중증도 등에 따라 그 범위를 달리하여 투여할 수 있다. 본 발명의 약학적 조성물을 투여하는 경우 일일 투여량은 약 0.01 내지 1000 ㎎/㎏ 이고, 보다 구체적으로 0.1 내지 100 ㎎/㎏ 이며, 일일 일 회 내지 수 회에 나누어 투여할 수 있다.The pharmaceutical composition of the present invention can be administered orally or parenterally according to a desired method, and the dosage depends on the patient's weight, age, health condition, diet, administration time, administration method, disease type and severity of disease, etc. It can be administered by varying the range. When administering the pharmaceutical composition of the present invention, the daily dose is about 0.01 to 1000 mg/kg, and more specifically, 0.1 to 100 mg/kg, and may be administered once to several times a day.
또한, 본 발명의 일 양태에서, 상기 대사성 질환의 예방 또는 치료용 약학적 조성물을 포함하는 경구용 제제를 제공한다. 구체적인 본 발명의 일 양태에서, 상기 경구용 제제는 정제, 과립제, 환제, 산제, 캅셀제 및 액제로 구성된 그룹으로부터 선택되는 것이다. In addition, in one aspect of the present invention, there is provided an oral preparation comprising a pharmaceutical composition for preventing or treating the metabolic disease. In a specific embodiment of the present invention, the oral preparation is selected from the group consisting of tablets, granules, pills, powders, capsules and liquids.
또한, 본 발명은 대사성 질환의 예방 또는 치료용 약학적 조성물을 포함하는 주사용 제제를 제공한다.In addition, the present invention provides an injectable formulation comprising a pharmaceutical composition for the prevention or treatment of metabolic diseases.
본 발명의 일 양태에서 경구 투여를 위한 약학적 제제는 예를 들어 정제, 과립제, 환제, 산제, 캅셀제 또는 액제와 같은 투여량 단위 형태 및 앰플로 존재할 수 있다. 이들은 그 자체로 공지된 방법, 예를 들어 통상적인 혼합, 입상화, 당제 조제, 용해 또는 동결건조 방법에 의해 제조된다.In one embodiment of the present invention, the pharmaceutical preparation for oral administration may be in the form of dosage units such as tablets, granules, pills, powders, capsules or liquids, and ampoules. They are prepared by methods known per se, for example by customary mixing, granulation, confection, dissolution or lyophilization methods.
다른 경구 투여 가능한 약학적 제제로는 캅셀제가 있으며, 젤라틴으로 만들어진 건식-충진 캅셀, 및 젤라틴 및 가소제, 예컨대 글리세롤 또는 소르비톨로 만들어진 연질의 밀봉된 캅셀제이다. 건식-충진 캅셀은 입자의 형태, 예를 들어 충진제, 예컨대 락토스, 결합제, 예컨대 전분, 및/또는 활택제, 예컨대 탈크 또는 스테아르산마그네슘, 및 적절하다면 안정화제와의 혼합물의 형태로 활성 성분을 함유할 수도 있다. 연질 캅셀에서, 활성 성분을 적절한 액체, 예컨대 지방 오일, 파라핀 오일 또는 액체 폴리에틸렌 글리콜에 용해 또는 현탁시키고, 여기에 안정화제를 첨가할 수도 있다.Other orally administrable pharmaceutical preparations include capsules, dry-filled capsules made of gelatin, and soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. Dry-filled capsules contain the active ingredient in the form of particles, for example in mixture with fillers such as lactose, binders such as starches, and/or glidants such as talc or magnesium stearate, and if appropriate stabilizers You may. In soft capsules, the active ingredient is dissolved or suspended in a suitable liquid, such as fatty oil, paraffin oil or liquid polyethylene glycol, to which a stabilizing agent may also be added.
본 발명의 제형이 액제인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the present invention is a liquid formulation, a solvent, solubilizer or emulsifier is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- butyl glycol oil, glycerol fatty ester, polyethylene glycol or fatty acid ester of sorbitan.
본 발명의 일 양태에서, 비경구 투여는 이에 제한되는 것은 아니나 예를 들어, 정맥 내, 피하, 피내, 복강내 또는 국소에 적용할 수 있다. 비경구 투여를 위한 약학적 제제는 살균될 수 있고/있거나 보존제, 안정화제, 습윤제 및/또는 유화제, 용해제, 삼투압을 조절하기 위한 염 및/또는 완충액과 같은 첨가제를 함유할 수 있다.In one embodiment of the present invention, parenteral administration may be, but is not limited to, for example, intravenous, subcutaneous, intradermal, intraperitoneal or topical application. Pharmaceutical preparations for parenteral administration may be sterile and/or contain additives such as preservatives, stabilizers, wetting and/or emulsifying agents, solubilizing agents, salts for adjusting the osmotic pressure and/or buffers.
또한, 본 발명 약학적 조성물은 비알코올성 지방간 질환(non-alcoholic fatty liver disease; NAFLD) 치료제와 병용 투여할 수 있다.In addition, the pharmaceutical composition of the present invention may be administered in combination with a therapeutic agent for non-alcoholic fatty liver disease (NAFLD).
또한, 본 발명 약학적 조성물은 비알코올성 지방간염(non-alcoholic steatohepatitis; NASH) 치료제와 병용 투여할 수 있다.In addition, the pharmaceutical composition of the present invention may be administered in combination with a therapeutic agent for non-alcoholic steatohepatitis (NASH).
본 발명의 일 양태에서, 상기 비알코올성 지방간 질환 치료제는 OCA (Obeticholic acid), 티아졸리딘디온(Thiazolidinediones), 비타민 E, 메트포르민, 스타틴, 우르소디옥시콜산(Ursodeoxycholic acid), 불포화 지방산, 안지오텐신 수용체 차단제, 펜톡시필린(Pentoxifylline), GLP-1 수용체 아고니스트(Glucagon-like peptide 1 receptor agonists), DPP-4 억제제(Dipeptidyl peptidase 4 inhibitors), SGLT2 억제제(sodium/glucose cotransporter 2 inhibitors), 엘라피브라노(Elafibranor) 및 텔미사르탄(Telmisartan)으로 이루어지는 군으로부터 선택되는 어느 하나 이상이다.In one embodiment of the present invention, the therapeutic agent for nonalcoholic fatty liver disease is OCA (Obeticholic acid), thiazolidinediones, vitamin E, metformin, statin, ursodeoxycholic acid, unsaturated fatty acid, angiotensin receptor blocker , Pentoxifylline, GLP-1 receptor agonists (Glucagon-like peptide 1 receptor agonists), DPP-4 inhibitors (Dipeptidyl peptidase 4 inhibitors), SGLT2 inhibitors (sodium/glucose cotransporter 2 inhibitors), elafibrano (Elafibranor) and telmisartan (Telmisartan) is any one or more selected from the group consisting of.
또한, 본 발명은 소듐 타우로디옥시콜레이트(sodium taurodeoxycholate), 이의 용매화물, 또는 이의 식품학적으로 허용가능한 염을 포함하는, 대사성 질환의 예방 또는 개선용 식품 조성물에 관한 것이다.In addition, the present invention relates to a food composition for preventing or improving metabolic diseases, comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof.
또한, 본 발명은 소듐 타우로디옥시콜레이트(sodium taurodeoxycholate), 이의 용매화물, 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 질환의 치료 방법을 제공한다.In addition, the present invention provides a method for treating a metabolic disease comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
이하, 본 발명을 실시예 및 실험예에 의해 상세히 설명한다.Hereinafter, the present invention will be described in detail by way of Examples and Experimental Examples.
단, 하기 실시예 및 실험예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예 및 실험예에 한정되는 것은 아니다.However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.
<실시예 1> 시료의 제조<Example 1> Preparation of samples
소듐 타우로디옥시콜레이트는 New Zealand Parmaceuticals Ltd. (Palmerston North, 뉴질랜드)로부터 구매하였다. Telmisartan(Micardis®)은 Boehringer Ingelheim GmbH(독일)로부터 구매하여, 정제수에 용해시켰다. DPBS(Dulbecco’s Phosphate-Buffered Salines)는 WelGene Inc., (LB001-02, 대한민국)으로부터 구매하여, 비히클 화합물(vehicle compound)로 사용하였다.Sodium Taurodeoxycholate is manufactured by New Zealand Pharmaceuticals Ltd. (Palmerston North, New Zealand). Telmisartan (Micardis®) was purchased from Boehringer Ingelheim GmbH (Germany) and dissolved in purified water. DPBS (Dulbecco's Phosphate-Buffered Salines) was purchased from WelGene Inc., (LB001-02, Korea) and used as a vehicle compound.
<실험예 1> 비알코올성 지방간염 동물모델 및 시험 디자인<Experimental Example 1> Non-alcoholic steatohepatitis animal model and test design
<1-1> 비알코올성 지방간염 동물모델<1-1> Nonalcoholic steatohepatitis animal model
소듐 타우로디옥시콜레이트의 비알코올성 지방간염에 대한 치료효과 확인을 위해 마우스 STAM 모델을 준비하였다. To confirm the therapeutic effect of sodium taurodioxycholate on nonalcoholic steatohepatitis, a mouse STAM model was prepared.
STAM 모델은 C57BL/6 마우스에 고지방식을 통한 식이요법과 화학물질 스트렙토조토신(streptozotocin)의 조합을 통해 유발되었다. C57BL/6 마우스(14일 임신 암컷)는 Japan SLC, Inc.(일본)으로부터 입수하였고, STAM 모델은 136 C57BL/6 수컷 마우스로 SMC Laboratories, Inc.(도쿄, 일본)에서 수행되었다. 연구에서 사용된 모든 동물은 일본 약리학회동물 동물 사용 지침서에 따라 수용 및 관리되었다. 구체적으로, 비알코올성 지방간염(NASH)은 생후 2일 후 200 μg 스트렙토조토신(STZ, Sigma-Aldrich, 미국) 용액을 단일 피하 주사하고, 4주령 후 고지방식(HFD, 57 kcal%fat, CLEA Japan, Cat# HFD32, 일본)을 자유롭게 제공하여 유발하였다. 마우스는 무작위로 할당되었으며, 구체적인 연구 설계는 도 1a에 나타내었다.The STAM model was induced in C57BL/6 mice through a combination of a high-fat diet and the chemical streptozotocin. C57BL/6 mice (14 days pregnant female) were obtained from Japan SLC, Inc. (Japan), and the STAM model was performed at SMC Laboratories, Inc. (Tokyo, Japan) with 136 C57BL/6 male mice. All animals used in the study were housed and managed according to the Animal Use Guidelines of the Japanese Pharmacological Association. Specifically, nonalcoholic steatohepatitis (NASH) was treated with a single subcutaneous injection of 200 μg streptozotocin (STZ, Sigma-Aldrich, USA) solution 2 days after birth, and a high-fat diet (HFD, 57 kcal%fat, CLEA) after 4 weeks of age. Japan, Cat# HFD32, Japan) was induced free of charge. Mice were randomly assigned, and the specific study design is shown in Figure 1a.
6주령에, 비히클 화합물인 DPBS, TDCA(Taurodeoxycholic acid) 및 텔미사르탄(Telmisartan) 각각을 매일 경구 투여하여, 9주령까지 치료하였다. 또한, 각각의 치료전에 체중을 측정하였다. 그 후, 이소플루란(isoflurane)(Pfizer Inc., 미국) 마취 하에 직접 심장천자(direct cardiac puncture)를 통해 9주령 마우스를 방혈시켜 안락사시켰다.At 6 weeks of age, vehicle compounds DPBS, TDCA (Taurodeoxycholic acid), and telmisartan, respectively, were orally administered daily, and treatment was performed until 9 weeks of age. In addition, body weight was measured before each treatment. Thereafter, 9-week-old mice were euthanized by exsanguination by direct cardiac puncture under isoflurane (Pfizer Inc., USA) anesthesia.
<1-2> 조직학적 분석<1-2> Histological analysis
1-2-1. H&E 염색(staining)을 위해, 부인용액(Bouin’s solution)으로 고정된 간 조직의 파라핀 블록으로부터 절편을 절단하고, Lillie-Mayer’s Hematoxylin(Muto Pure Chemicals Co. Ltd., 일본) 및 에오신 용액(FUJIFILM Wako Pure Chemical Corporation, 일본)으로 염색하였다.1-2-1. For H&E staining, sections were cut from paraffin blocks of liver tissue fixed with Bouin's solution, and Lillie-Mayer's Hematoxylin (Muto Pure Chemicals Co. Ltd., Japan) and Eosin solution (FUJIFILM Wako Pure) Chemical Corporation, Japan).
1-2-2. NAFLD 활동 점수(NAFLD Activity score, NAS)는 Kleiner 기준에 따라 계산되었다.1-2-2. NAFLD activity score (NAFLD activity score, NAS) was calculated according to the Kleiner criteria.
1-2-3. 콜라겐 침착의 시각화를 위해, 부인 고정된 간 절편을 피크로-시리우스 레드 용액(picro-Sirius red solution)(Waldeck, 독일)을 사용하여 염색하였다.1-2-3. For visualization of collagen deposition, denial fixed liver sections were stained using picro-Sirius red solution (Waldeck, Germany).
1-2-4. 섬유증(fibrosis) 영역의 정량적 분석을 위해, 시리우스 레드로 염색된 절편을 200배 배율 디지털카메라(DFC295; Leica, 독일)를 사용하여 중심정맥 주위에서 포착하였다. 또한, 5개 필드/섹션에서 양성 영역은 Image J 소프트웨어(National Institue of Health, 미국)을 사용하여 측정되었다.1-2-4. For quantitative analysis of the fibrosis area, a section stained with Sirius Red was captured around the central vein using a digital camera (DFC295; Leica, Germany) at 200x magnification. In addition, positive areas in 5 fields/section were measured using Image J software (National Institute of Health, USA).
1-2-5. 면역 조직 화학을 위해, Tissue-Tek O.C.T 화합물에 고정된 냉동 간 조직으로부터 절편을 절단하고, 아세톤에 고정시켰다. 내인성 퍼옥시다아제(Endogenous peroxidase) 활성을 0.03% H2O2를 10분 동안 사용하여 차단하고, 그 후 Block Ace(DS pharma Biomedical Co. Ltd., 일본)과 함께 10분 동안 인큐베이션하였다. 상기 절편을 실온에서 1시간동안 망상 섬유 아세포 항체(reticular fibroblast antibodis)(BMA Biomedicals Inc., Cat#: T-2109, Clone: ER-TR7, 스위스)와 함께 배양하였다. 이어서, 절편을 비오틴 접합된 이차 항체(biotin-conjugated secondary antibody)(VECTASTAIN Elite ABC kit, Vector Laboratories, Cat#: PK-4004, 미국)과 함께 배양하고, 그 후 실온에서 ABC reagent와 함께 30분동안 배양하였다.1-2-5. For immunohistochemistry, sections were excised from frozen liver tissue fixed in Tissue-Tek OCT compound and fixed in acetone. Endogenous peroxidase activity was blocked using 0.03% H 2 O 2 for 10 minutes, and then incubated with Block Ace (DS Pharma Biomedical Co. Ltd., Japan) for 10 minutes. The sections were incubated with reticular fibroblast antibodis (BMA Biomedicals Inc., Cat#: T-2109, Clone: ER-TR7, Switzerland) for 1 hour at room temperature. Then, the fragment was incubated with a biotin-conjugated secondary antibody (VECTASTAIN Elite ABC kit, Vector Laboratories, Cat#: PK-4004, USA), and then incubated with ABC reagent at room temperature for 30 minutes. cultured.
효소 기질 반응은 3,3′-디아미노벤지딘/H2O2 용액(3,3’-diaminobenzidine/H2O2 solution)(Nichirei Bioscience inc., 일본)을 사용하여 수행하였다.The enzyme substrate reaction was performed using a 3,3′-diaminobenzidine/H 2 O 2 solution (3,3′-diaminobenzidine/H 2 O 2 solution) (Nichirei Bioscience inc., Japan).
1-2-6. ER-TR7 양성 영역의 정량 분석을 위해, 200배 배율 디지털카메라(DFC295; Leica)를 사용하여 중심정맥 주위에서 ER-TR7 면역 염색 섹션의 브라이트 필드 이미지를 캡쳐하였다. 또한, 5개 필스/섹션에서 양성 영역은 Image J 소프트웨어(National Institue of Health, 미국)을 사용하여 측정되었다.1-2-6. For quantitative analysis of ER-TR7 positive regions, bright field images of ER-TR7 immunostained sections were captured around the central vein using a 200x digital camera (DFC295; Leica). In addition, positive areas in 5 fields/section were measured using Image J software (National Institute of Health, USA).
<1-3> 정량적 RT-PCR(Real Time Polymerase Chain Reaction)<1-3> Quantitative RT-PCR (Real Time Polymerase Chain Reaction)
총 RNA는 제조사의 지시에 따라 RNAiso(Takara Bio, 일본)을 사용하여 간 샘플로부터 추출되었다. 구체적으로, 1 μg의 RNA를 반응 혼합물 20 μL를 포함시켜 사용하여 역전사하였으며, 반응 혼합물은 4.4 mM MgCl2(F. Hoffmann-La Roche, 스위스), 40 U RNase 억제제(Toyobo, 일본), 0.5 mM dNTP(Proemga, 미국), 6.28 μM 랜덤 헥사머(Promega, 미국), 5x first strand buffer(Promega, 미국), 10 mM 디티오트레이톨(dithiothreitol)(Invitrogen, 미국), 200 U MMLV-RT(Invitrogen, 미국)를 포함한다. 반응은 37 ℃에서 1시간 진행시키고, 그 후 99 ℃에서 5분간 진행시켰다.Total RNA was extracted from liver samples using RNAiso (Takara Bio, Japan) according to the manufacturer's instructions. Specifically, 1 μg of RNA was reverse transcribed using 20 μL of the reaction mixture, and the reaction mixture was 4.4 mM MgCl 2 (F. Hoffmann-La Roche, Switzerland), 40 U RNase inhibitor (Toyobo, Japan), 0.5 mM dNTP (Proemga, USA), 6.28 μM random hexamer (Promega, USA), 5x first strand buffer (Promega, USA), 10 mM dithiothreitol (Invitrogen, USA), 200 U MMLV-RT (Invitrogen) , the United States). The reaction was carried out at 37 °C for 1 hour, and then at 99 °C for 5 minutes.
특정 유전자 발현은 PowerUp™ SYBR™ Green Master Mix (Applied Biosystems Inc., 미국, Cat # A25779)을 사용하여 real-time PCR을 통해 정량화되었으며, Quantstudio6 Flex System Instrument (Applied Biosystems Inc., 미국)을 사용하여 수행되었다.Specific gene expression was quantified via real-time PCR using PowerUp ™ SYBR ™ Green Master Mix (Applied Biosystems Inc., USA, Cat # A25779), and Quantstudio6 Flex System Instrument (Applied Biosystems Inc., USA) using carried out
60S 산성 리보솜 단백질 P0(RPLP0)RNA 발현은 내부 대조군으로 측정하였다.60S acidic ribosomal protein P0 (RPLP0) RNA expression was measured as an internal control.
표적 유전자의 상대 발현 수준을 RPLP0에 대해 정규화한 후 비교하였다. 상대 정량은 △Ct method를 사용하여 수행되었다.Relative expression levels of target genes were compared after normalization to RPLP0. Relative quantification was performed using the ΔCt method.
프라이머 서열은 다음과 같다: CCL2(MCP1) (Forward: 5’- AAACTGCATCTGCCCTAAGG-3’ / Reverse: 5’-AGAAGTGCTTGAGGTGGTTG-3’).The primer sequence is as follows: CCL2(MCP1) (Forward: 5'-AAACTGCATCTGCCCTAAGG-3' / Reverse: 5'-AGAAGTGCTTGAGGTGGTTG-3').
<1-4> ELISA<1-4> ELISA
상기 마우스 혈장에서 TNF-α은 마우스 TNF-α 비코팅 ELISA Kit(Invitrogen, Cat# 88-7324, 미국), TIMP1은 마우스 TIMP1 ELSIA Kit(abcam, Cat# ab196265, Cambridge, 영국)을 통해 분석하였으며, 키트는 제조업체의 지침에 따라 사용하였다.TNF-α in the mouse plasma was analyzed using the mouse TNF-α uncoated ELISA Kit (Invitrogen, Cat# 88-7324, USA), and TIMP1 was analyzed through the mouse TIMP1 ELSIA Kit (abcam, Cat# ab196265, Cambridge, UK), The kit was used according to the manufacturer's instructions.
<1-5> 데이터 및 통계분석<1-5> Data and Statistical Analysis
데이터는 평균 ±SD로 표시되며, 두 개 이상의 그룹이 포함된 모든 데이터는 일원 분산분석(One-way ANOVA)에 의해 분석되었고, 그 후 Dunnett 또는 Tukey 다중 비교 테스트, 또는 Kruskal Wallis 테스트에 의해 분석되며, 그 후 Dunn 다중 비교 테스트에 의해 분석되었다. 도면에서 별표는 *p<0.05, **p<0.01, ***p<0.001 및 ****p<0.0001을 의미한다. 분석은 GraphPad Prism 8.3.1 (San Diego, CA, 미국)을 통해 수행되었다.Data are presented as mean ± SD, all data with two or more groups were analyzed by one-way ANOVA, followed by Dunnett or Tukey multiple comparison test, or Kruskal Wallis test. , which were then analyzed by the Dunn multiple comparison test. In the figure, asterisks mean *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001. Analysis was performed via GraphPad Prism 8.3.1 (San Diego, CA, USA).
<실험예 2> 비알코올성 지방간염 치료효과 확인<Experimental Example 2> Confirmation of treatment effect for nonalcoholic steatohepatitis
<2-1> 몸 및 간 무게 변화 확인<2-1> Check body and liver weight changes
상기 <실험예 1>에 따른 실험방법에 의한 비알코올성 지방 간염의 STAM 모델에서 TDCA의 효과를 조사하였다. 안지오텐신 II 수용체 길항제(angiotensin II receptor antagonist) 및 페록시좀 증식제 활성화 수용체-γ 작용제(peroxisome proliferator-activated receptor-γ agonist)인 텔미사르탄(Telmisartan)은 섬유증(fibrosis) 및 염증(inflammation)을 감소시켜 NASH를 개선하는 특성을 가지고 있다. 본 발명의 실험에서 텔미사르탄은 기준 약물로 사용하였다. The effect of TDCA was investigated in the STAM model of nonalcoholic steatohepatitis by the experimental method according to <Experimental Example 1>. Telmisartan, an angiotensin II receptor antagonist and peroxisome proliferator-activated receptor-γ agonist, reduces fibrosis and inflammation It has the characteristic of improving NASH. In the experiment of the present invention, telmisartan was used as a reference drug.
STAM 모델에서 TDCA와 텔미사르탄은 치료효과가 있는 것을 확인하였다(도 1A).In the STAM model, it was confirmed that TDCA and telmisartan had therapeutic effects (FIG. 1A).
처리군(TDCA)과 비히클 군(DPBS) 사이에 체중에 대한 유의미한 영향이 관찰되지 않아 일반적 약물 독성에 의한 체중감소는 없는 것으로 확인하였다. 다만, 텔미사르탄 군이 비히클 군과 비교하여 현저한 감소가 발생하는 것을 확인하였다(도 1B).No significant effect on body weight was observed between the treatment group (TDCA) and the vehicle group (DPBS), so it was confirmed that there was no weight loss due to general drug toxicity. However, it was confirmed that a significant decrease occurred in the telmisartan group compared to the vehicle group (FIG. 1B).
처리군(TDCA)과 비히클 군(DPBS) 사이에 간 대 체중 비율에 대한 유의미한 영향이 관찰되지 않았다. 다만, 텔미사르탄 군이 비히클 군과 비교하여 현저한 감소가 발생하는 것을 확인하였다(도 1C). 텔미사르탄 군은 종래 보고와 같이 동물군에서 간 대 체중 비율을 감소시켰다.No significant effect on liver to body weight ratio was observed between the treatment group (TDCA) and the vehicle group (DPBS). However, it was confirmed that a significant decrease occurred in the telmisartan group compared to the vehicle group ( FIG. 1C ). The telmisartan group decreased the liver-to-body weight ratio in the animal group as previously reported.
<2-2> 간의 조직학적 분석<2-2> Histological analysis of liver
STAM 모델 그룹은 정상 그룹과 비교하여 간세포 지방증(hepatocyte steatosis), 벌루닝(ballooning) 및 염증 세포 침윤(inflammatory cell infiltration)이 발생하였다(도 2A).Compared to the normal group, the STAM model group had hepatocyte steatosis, ballooning, and inflammatory cell infiltration (FIG. 2A).
비히클 군(DPBS)과 비교하여 TDCA 1.25 mg/kg 처리군 및 텔미사르탄 군은 총 NAS는 유의적으로 감소한 것을 확인하여 NASH의 중요한 임상 증상 척도 NAS 병변지수의 개선을 확인하였다(도 2B).Compared to the vehicle group (DPBS), the TDCA 1.25 mg/kg treatment group and the telmisartan group showed a significant decrease in total NAS, confirming the improvement of the NAS lesion index, an important clinical symptom measure of NASH ( FIG. 2B ).
<실험예 3> 섬유증(Fibrosis) 치료효과 확인<Experimental Example 3> Confirmation of treatment effect on fibrosis
STAM 마우스에서 시리우스 레드로 염색된 간 절편, 및 항 섬유 아세포 항체 ER-TR7 염색된 간 절편은 정상 생쥐와 비교하여 간엽 소엽 영역(hepatic lobule region)에서 콜라겐 침착이 증가하는 것으로 나타났다(도 3A).Liver sections stained with Sirius Red and anti-fibroblast antibody ER-TR7 stained liver sections from STAM mice showed increased collagen deposition in the hepatic lobule region compared to normal mice (Fig. 3A).
비히클 군(DPBS)과 비교하여, TDCA 2.5 mg/kg 처리군 및 텔미사르탄 군에서 혈청 양성 영역에서 섬유증 면적의 유의미한 감소가 있어 NASH 치료약물의 중요한 임상 치료 척도인 섬유증에 대해 치료 효과를 확인하였다(도 3B).Compared with the vehicle group (DPBS), there was a significant decrease in the area of fibrosis in the seropositive area in the TDCA 2.5 mg/kg treatment group and the telmisartan group, confirming the therapeutic effect on fibrosis, an important clinical treatment measure of NASH drugs. (Fig. 3B).
또한, 비히클 군(DPBS)과 비교하여, TDCA 10 mpk 군에서 ER-TR7 양성 면적이 유의미하게 감소한 것을 확인하여 섬유증의 개선이 간조직내 섬유아세포를 억제하는 효과와 상관성이 있음을 확인하였다(도 3B).In addition, compared with the vehicle group (DPBS), it was confirmed that the ER-TR7 positive area was significantly reduced in the TDCA 10 mpk group, confirming that the improvement of fibrosis was correlated with the effect of inhibiting fibroblasts in the liver tissue (Fig. 3B).
<실험예 4> 염증성, 콜라겐/섬유증 마커의 유전자 및 단백질 발현<Experimental Example 4> Inflammatory, collagen / fibrosis marker gene and protein expression
NASH에서 염증(inflammation) 및 섬유증(fibrosis)과 관련된 마우스 간 조직 유전자의 mRNA 발현 수준을 상기 실험예에 따라 분석하였다. 분석된 염증 마커는 MCP-1(monocyte chemoattractant protein-1)이다. 분석결과, MCP-1에서 TDCA 2.5 mg/kg 처리군이 비히클 군(DPBS)과 비교하여 현저한 감소가 관찰되어 항염증 효과를 확인하였다(도 4A).The mRNA expression level of mouse liver tissue genes related to inflammation and fibrosis in NASH was analyzed according to the above experimental example. The analyzed inflammatory marker is monocyte chemoattractant protein-1 (MCP-1). As a result of the analysis, a significant decrease was observed in the TDCA 2.5 mg/kg treatment group compared to the vehicle group (DPBS) in MCP-1, confirming the anti-inflammatory effect (FIG. 4A).
또한, ELISA에 의해 마우스 혈장에서 염증성 사이토 카인 TNF-α 및 섬유증 마커 TIMP1의 단백질 발현 수준을 분석하였다. 분석결과, TDCA 2.5 mg/kg 처리군이 비히클 군(DPBS)과 비교하여 TNF-α의 현저한 감소가 관찰되어 중요 염증성 사이토카인을 억제하는 항염증 효과가 있음을 확인하였다(도 4B).In addition, protein expression levels of the inflammatory cytokine TNF-α and the fibrosis marker TIMP1 were analyzed in mouse plasma by ELISA. As a result of the analysis, the TDCA 2.5 mg/kg treatment group showed a significant decrease in TNF-α compared to the vehicle group (DPBS), confirming that there was an anti-inflammatory effect of suppressing important inflammatory cytokines (FIG. 4B).
본 발명은 대사성 질환의 예방 또는 치료, 보다 구체적으로는 비알코올성 지방간염(NASH)의 예방 또는 치료에 유용하게 이용할 수 있다.The present invention can be usefully used for the prevention or treatment of metabolic diseases, more specifically, the prevention or treatment of nonalcoholic steatohepatitis (NASH).

Claims (14)

  1. 소듐 타우로디옥시콜레이트(sodium taurodeoxycholate), 이의 용매화물, 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 질환의 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating metabolic diseases, comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  2. 제1항에 있어서, According to claim 1,
    상기 대사성 질환은 비알코올성 지방간 질병(NAFLD), 비알코올성 지방간염(NASH), 간 섬유화(Liver Fibrosis), 비만, 당뇨병 및 고지혈증으로 구성된 그룹으로부터 선택된 1종 이상인, 약학적 조성물.The metabolic disease is at least one selected from the group consisting of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis (Liver Fibrosis), obesity, diabetes and hyperlipidemia, a pharmaceutical composition.
  3. 제1항에 있어서,According to claim 1,
    상기 대사성 질환은 비알코올성 지방간염(NASH)인, 약학적 조성물.The metabolic disease is nonalcoholic steatohepatitis (NASH), a pharmaceutical composition.
  4. 제1항에 있어서,According to claim 1,
    상기 대사성 질환은 간 섬유화(Liver Fibrosis)인, 약학적 조성물.The metabolic disease is liver fibrosis (Liver Fibrosis), a pharmaceutical composition.
  5. 제1항에 있어서, According to claim 1,
    상기 조성물은 지방증(steatosis), 염증(inflammation) 또는 벌룬 현상(ballooning)을 감소시키는 것인, 약학적 조성물.The composition is to reduce the steatosis (steatosis), inflammation (inflammation) or ballooning phenomenon (ballooning), the pharmaceutical composition.
  6. 제1항에 있어서, According to claim 1,
    상기 조성물은 섬유화(fibrosis) 또는 경화(cirrhosis)를 감소시키는 것인, 약학적 조성물.The composition is to reduce fibrosis or hardening (cirrhosis), a pharmaceutical composition.
  7. 제1항 내지 제6항 중 어느 한 항에 따른 약학적 조성물을 포함하는 경구용 제제.An oral formulation comprising the pharmaceutical composition according to any one of claims 1 to 6.
  8. 제7항에 있어서,8. The method of claim 7,
    상기 경구용 제제는 정제, 과립제, 환제, 산제, 캅셀제 및 액제로 구성된 그룹으로부터 선택되는 것인 경구용 제제.The oral preparation is an oral preparation selected from the group consisting of tablets, granules, pills, powders, capsules and liquids.
  9. 제1항 내지 제6항 중 어느 한 항에 따른 약학적 조성물을 포함하는 주사용 제제.An injectable formulation comprising the pharmaceutical composition according to any one of claims 1 to 6.
  10. 제1항에 있어서, According to claim 1,
    상기 조성물은 비알코올성 지방간 질환(non-alcoholic fatty liver disease; NAFLD) 치료제와 병용 투여하는 것인, 약학적 조성물.The composition is non-alcoholic fatty liver disease (non-alcoholic fatty liver disease; NAFLD) will be administered in combination with a therapeutic agent, the pharmaceutical composition.
  11. 제1항에 있어서, According to claim 1,
    상기 조성물은 비알코올성 지방간염(non-alcoholic steatohepatitis; NASH) 치료제와 병용 투여하는 것인, 약학적 조성물.The composition is non-alcoholic steatohepatitis (non-alcoholic steatohepatitis; NASH) will be administered in combination with a therapeutic agent, the pharmaceutical composition.
  12. 제10항에 있어서, 11. The method of claim 10,
    상기 비알코올성 지방간 질환 치료제는 OCA (Obeticholic acid), 티아졸리딘디온(Thiazolidinediones), 비타민 E, 메트포르민, 스타틴, 우르소디옥시콜산(Ursodeoxycholic acid), 불포화 지방산, 안지오텐신 수용체 차단제, 펜톡시필린(Pentoxifylline), GLP-1 수용체 아고니스트(Glucagon-like peptide 1 receptor agonists), DPP-4 억제제(Dipeptidyl peptidase 4 inhibitors), SGLT2 억제제(sodium/glucose cotransporter 2 inhibitors), 엘라피브라노(Elafibranor) 및 텔미사르탄(Telmisartan)으로 이루어지는 군으로부터 선택되는 어느 하나 이상인, 약학적 조성물.The nonalcoholic fatty liver disease treatment agent is OCA (Obeticholic acid), thiazolidinediones, vitamin E, metformin, statin, ursodeoxycholic acid, unsaturated fatty acid, angiotensin receptor blocker, pentoxifylline , GLP-1 receptor agonists (Glucagon-like peptide 1 receptor agonists), DPP-4 inhibitors (Dipeptidyl peptidase 4 inhibitors), SGLT2 inhibitors (sodium/glucose cotransporter 2 inhibitors), Elafibranor and Telmisartan (Telmisartan) Any one or more selected from the group consisting of, a pharmaceutical composition.
  13. 소듐 타우로디옥시콜레이트(sodium taurodeoxycholate), 이의 용매화물, 또는 이의 식품학적으로 허용가능한 염을 포함하는, 대사성 질환의 예방 또는 개선용 식품 조성물.Sodium taurodeoxycholate (sodium taurodeoxycholate), a solvate thereof, or a food composition comprising a pharmaceutically acceptable salt thereof, for the prevention or improvement of metabolic diseases food composition.
  14. 소듐 타우로디옥시콜레이트(sodium taurodeoxycholate), 이의 용매화물, 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 질환의 치료 방법.A method of treating a metabolic disease comprising sodium taurodeoxycholate, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
PCT/KR2021/011231 2020-09-18 2021-08-23 Pharmaceutical composition for treating nonalcoholic steatohepatitis and liver fibrosis WO2022059946A1 (en)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2633859A1 (en) * 2007-05-14 2013-09-04 Seoul National University Industry Foundation Use of biological surfactant as anti-inflammatory agent and tissue preservative solution
US20170071968A1 (en) * 2008-03-04 2017-03-16 Alexander Vladimirovich Dikovskiy Pharmaceutical composition based on a hepatoprotector and prebiotic, and method for administrating
WO2019161025A1 (en) * 2018-02-14 2019-08-22 Lumos Pharma, Inc. Compositions for the treatment of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis
KR20200091592A (en) * 2019-01-23 2020-07-31 서울대학교산학협력단 Composition for preventing or treating of inflammatory bowel disease comprising taurodeoxycholic acid or pharmaceutically acceptable salts thereof as an active ingredient

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2633859A1 (en) * 2007-05-14 2013-09-04 Seoul National University Industry Foundation Use of biological surfactant as anti-inflammatory agent and tissue preservative solution
US20170071968A1 (en) * 2008-03-04 2017-03-16 Alexander Vladimirovich Dikovskiy Pharmaceutical composition based on a hepatoprotector and prebiotic, and method for administrating
WO2019161025A1 (en) * 2018-02-14 2019-08-22 Lumos Pharma, Inc. Compositions for the treatment of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis
KR20200091592A (en) * 2019-01-23 2020-07-31 서울대학교산학협력단 Composition for preventing or treating of inflammatory bowel disease comprising taurodeoxycholic acid or pharmaceutically acceptable salts thereof as an active ingredient

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
YAHAGHI LEYLA, EBRAHIM‐HABIBI AZADEH, HAYATI‐ROODBARI NASIM, IRANI SHIVA, YAGHMAEI PARICHEHREH: "A simple method for inducing nonalcoholic steatohepatitis with fibrosis", ANIMAL MODELS AND EXPERIMENTAL MEDICINE, vol. 2, no. 4, 1 December 2019 (2019-12-01), pages 282 - 290, XP055912015, ISSN: 2576-2095, DOI: 10.1002/ame2.12089 *

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