RU2803733C1 - Pharmaceutical composition for prevention or treatment of non-alcoholic steathoepatitis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions relates to the treatment of non-alcoholic fatty infiltration of the liver. The pharmaceutical composition contains the following components:

, where A is an oxadiazole group, B is a pyrimidine group, substituted with a C1-C6 linear or branched alkyl group, X – F; at least one substance selected from pioglitazone, lobeglitazone, elafibranor, lanifibranor, saroglitazar, sitagliptin, vildagliptin, saxagliptin, linagliptin, hemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, dut ogliptin, firsocostat, obeticholic acid, tropifexor, 3-(2,6-dichlorophenyl)-4-(3'-carboxy-2-chlorostilben-4-yl)oxymethyl-5-isopropylisoxazole and cenicriviroc. Also the following is disclosed: the use of a pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease and for the preparation of a medicinal product, a method of the prevention or treatment of non-alcoholic fatty liver disease.

EFFECT: group of inventions improves lipid metabolism, reduces fat accumulation in liver tissue and suppresses histological damage.

14 cl, 13 dwg, 3 ex

Description

Field of technology to which the invention relates

The present invention relates to a pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease comprising a G protein-coupled receptor 119 (GPR119) agonist as an active ingredient and at least one drug with a different mechanism of action.

State of the art

Fatty liver disease is a pathological condition in which triglycerides accumulate excessively in hepatocytes, and is medically defined as a condition in which triglycerides constitute 5% or more of the liver's weight. Fatty liver is divided into alcoholic and non-alcoholic fatty liver depending on whether fatty liver is caused or not caused by excessive alcohol consumption. Non-alcoholic fatty liver disease (NAFLD) is a group of diseases that include all aspects of disease ranging from non-alcoholic fatty liver disease to steatohepatitis and cirrhosis. Simple non-alcoholic fatty liver disease is a disease in which only fat deposition occurs in the liver, without any evidence of hepatocyte damage and fibrosis, and fat deposition in the liver tissue increases only due to insulin resistance, etc. Non-alcoholic fatty liver disease may progress with transition to non-alcoholic steatohepatitis (NASH), accompanied by fibrosis due to damage to hepatocytes caused by an inflammatory response in accordance with oxidative stress, etc.

Unlike simple fatty liver, nonalcoholic steatohepatitis can progress to cirrhosis and liver cancer caused by irreversible liver damage, which increases liver disease-related and overall mortality (Hepatology, 2012(55):2005-2023). For this reason, according to statistics in the United States in 2013, non-alcoholic steatohepatitis was the second leading common cause of liver transplantation after hepatitis C. However, as of 2020, non-alcoholic steatohepatitis, rather than hepatitis C, was the first leading disease leading to liver transplantation and therefore there is an urgent need to develop a drug to prevent or treat non-alcoholic steatohepatitis (Clinical Gastroenterology and Hepatology, 2019(17):748-755).

Detailed Description of the Invention

Technical problem

One object of the present invention is a pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis, which includes the compound described by chemical formula 1, its pharmaceutically acceptable salts, its optical isomers, its hydrates, its solvates or mixtures thereof; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist.

The solution of the problem

A pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis in one embodiment of the present invention may include a compound represented by the following chemical formula 1, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof, or mixtures thereof; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist.

[Formula 1]

In the above formula 1, A may be an oxadiazole group, a dihydrooxazole group, a thiazole group, or a thiadiazole group. The above A may be unsubstituted or substituted by at least one substituent selected from the group consisting of a halogen atom, a C1-C6 straight or branched chain alkyl group, and a C1-C6 straight or branched chain hydroxyalkyl group. The alkyl group or hydroxyalkyl group may each independently be unsubstituted or substituted by a halogen atom or a C1-C6 alkoxy group.

B may be a pyridine group, a pyrimidine group, a pyrazine group or an oxadiazole group. The above B may be unsubstituted or substituted by at least one substituent selected from the group consisting of: halogen atom, C1-C6 straight or branched chain alkyl group and C1-C6 straight or branched hydroxyalkyl group, C1-C6 alkoxy group and oxadiazole group. A C1-C6 straight or branched chain alkyl group, a C1-C6 straight or branched hydroxyalkyl group, a C1-C6 alkoxy group or an oxadiazole group can each independently be unsubstituted or substituted with halogen, a C1-C6 alkyl group or a C1-C6 alkoxy group.

X can mean F, Cl, Br or I.

The above A could mean , , , or .

R ₁ - R ₃ , R ₅ and R ₆ can each independently represent at least one substituent selected from the group consisting of the following: a hydrogen atom, a halogen atom, a C1-C6 straight or branched chain alkyl group and a C1-C6 straight-chain alkyl group linear or branched chain hydroxyalkyl group. The alkyl group or hydroxyalkyl group may each independently be unsubstituted or substituted by a halogen atom or a C1-C6 alkoxy group.

The above B could mean , , , or .

The above R ₇ to R ₁₁ may each be independently substituted by at least one substituent selected from the group consisting of a hydrogen atom, a halogen atom, a C1-C6 straight or branched chain alkyl group, and a C1-C6 straight or branched chain alkyl group. chain hydroxyalkyl group, C1-C6 alkoxy group and oxadiazole group. The alkyl group, hydroxyalkyl group, alkoxy group or oxadiazole group can each independently be unsubstituted or substituted by a halogen atom, a C1-C6 alkyl group or a C1-C6 alkoxy group.

X could mean F.

In the above Formula 1, A above may be an oxadiazole group substituted with a C1-C6 straight or branched chain alkyl group, B above may be a pyrimidine group substituted with a C1-C6 straight or branched chain alkyl group, and X may be F .

In this embodiment, the term "halogen" may mean fluorine, chlorine, bromine or iodine.

In one embodiment, the term "alkyl" can mean a straight-chain or branched-chain hydrocarbon moiety unless otherwise indicated. An example of said C1-C6 alkyl may include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, hexyl, and the like.

In one embodiment, the term "alkoxy group" may include an alkyl oxygen radical containing alkyl as defined above unless otherwise noted. An example of said C1-C6 alkoxy group may include a methoxy group, an ethoxy group, a propoxy group, a butoxy group, a pentoxy group and the like.

In one embodiment, the term “heterocycle” or “heterocyclic” can mean a 5- to 13-membered heteroaromatic or non-aromatic compound containing from 1 to 3 heteroatoms selected from the group consisting of N, O and S, unless otherwise indicated.

In one embodiment, the compound represented by Formula 1 above may be at least one compound specifically selected from the group consisting of the following compounds:

2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-4,5-dihydrooxazole,

(R)-2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-4-methyl-4,5-dihydrooxazole,

(S)-2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-4-methyl-4,5-dihydrooxazole,

(S)-2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-methyl-4,5-dihydrooxazole,

(R)-2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-methyl-4,5-dihydrooxazole,

2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5,5-dimethyl-4,5-dihydrooxazole,

(R)-(2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-4,5-dihydrooxazol-5-yl )methanol,

(S)-(2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-4,5-dihydrooxazol-5-yl )methanol,

(R)-3-(2-(4-(3-(3,5-difluoro-4-(5-methyl-4,5-dihydrooxazol-2-yl)phenoxy)propyl)piperidin-1-yl)pyrimidine -5-yl)-5-isobutyl-1,2,4-oxadiazole,

(R)-5-(4-(3-(3,5-difluoro-4-(4-methyl-4,5-dihydrooxazol-2-yl)phenoxy)propyl)piperidin-1-yl)-3-isopropyl -1,2,4-oxadiazole,

(S)-5-(4-(3-(3,5-difluoro-4-(5-methyl-4,5-dihydrooxazol-2-yl)phenoxy)propyl)piperidin-1-yl)-3-isopropyl -1,2,4-oxadiazole,

5-(4-(3-(4-(5,5-dimethyl-4,5-dihydrooxazol-2-yl)-3,5-difluorophenoxy)propyl)piperidin-1-yl)-3-isopropyl-1, 2,4-oxadiazole,

3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-methyl-1,2,4-oxadiazole,

3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-propyl-1,2,4-oxadiazole,

3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole,

5-(tert-butyl)-3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-1,2,4- oxadiazole,

(3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-1,2,4-oxadiazol-5-yl)methanol ,

2-(3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-1,2,4-oxadiazol-5-yl )ethan-1-ol,

(S)-1-(3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-1,2,4-oxadiazole -5-yl)propan-1-ol,

(R)-1-(3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-1,2,4-oxadiazole -5-yl)propan-2-ol,

(S)-1-(3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-1,2,4-oxadiazole -5-yl)propan-2-ol,

2-(3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-1,2,4-oxadiazol-5-yl )-2-methylpropan-1-ol,

3-(2,6-difluoro-4-(3-(1-(5-propylpyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,2,4-oxadiazole,

3-(2,6-difluoro-4-(3-(1-(5-pentylpyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,2,4-oxadiazole,

3-(2,6-difluoro-4-(3-(1-(5-(trifluoromethyl)pyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,2,4- oxadiazole,

3-(2,6-difluoro-4-(3-(1-(5-methoxypyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,2,4-oxadiazole,

3-(2,6-difluoro-4-(3-(1-(5-isopropoxypyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,2,4-oxadiazole,

3-(4-(3-(1-(5-chloropyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole,

3-(4-(3-(1-(5-bromopyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole,

3-(2,6-difluoro-4-(3-(1-(5-(5-isobutyl-1,2,4-oxadiazol-3-yl)pyrimidin-2-yl)piperidin-4-yl)propoxy )phenyl)-5-methyl-1,2,4-oxadiazole,

3-(2,6-difluoro-4-(3-(1-(5-(5-isobutyl-1,2,4-oxadiazol-3-yl)pyrimidin-2-yl)piperidin-4-yl)propoxy )phenyl)-5-ethyl-1,2,4-oxadiazole,

3-(2,6-difluoro-4-(3-(1-(5-(5-isobutyl-1,2,4-oxadiazol-3-yl)pyrimidin-2-yl)piperidin-4-yl)propoxy )phenyl)-5-isopropyl-1,2,4-oxadiazole,

5-(sec-butyl)-3-(2,6-difluoro-4-(3-(1-(5-(5-isobutyl-1,2,4-oxadiazol-3-yl)pyrimidin-2-yl )piperidin-4-yl)propoxy)phenyl)-1,2,4-oxadiazole,

3-(2,6-difluoro-4-(3-(1-(5-(5-isobutyl-1,2,4-oxadiazol-3-yl)pyrimidin-2-yl)piperidin-4-yl)propoxy )phenyl)-5-(methoxymethyl)-1,2,4-oxadiazole,

(S)-1-(3-(2,6-difluoro-4-(3-(1-(5-(5-isobutyl-1,2,4-oxadiazol-3-yl)pyrimidin-2-yl) piperidin-4-yl)propoxy)phenyl)-1,2,4-oxadiazol-5-yl)propan-1-ol,

2-(3-(2,6-difluoro-4-(3-(1-(5-(5-isobutyl-1,2,4-oxadiazol-3-yl)pyrimidin-2-yl)piperidin-4- yl)propoxy)phenyl)-1,2,4-oxadiazol-5-yl)-2-methylpropan-1-ol,

3-(4-(3-(1-(5-chloropyrazin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole,

3-(2,6-difluoro-4-(3-(1-(5-(trifluoromethyl)pyridin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,2,4- oxadiazole,

3-(2,6-difluoro-4-(3-(1-(3-isopropyl-1,2,4-oxadiazol-5-yl)piperidin-4-yl)propoxy)phenyl)-5-methyl-1 ,2,4-oxadiazole,

3-(2,6-difluoro-4-(3-(1-(3-isopropyl-1,2,4-oxadiazol-5-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1 ,2,4-oxadiazole,

(3-(2,6-difluoro-4-(3-(1-(3-isopropyl-1,2,4-oxadiazol-5-yl)piperidin-4-yl)propoxy)phenyl)-1,2, 4-oxadiazol-5-yl)methanol,

2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-methyl-1,3,4-oxadiazole,

2-ethyl-5-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-1,3,4-oxadiazole,

2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,3,4-oxadiazole,

5-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-N-isopropyl-1,3,4-oxadiazol-2- amine,

2-(2,6-difluoro-4-(3-(1-(5-(trifluoromethyl)pyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-methyl-1,3,4- oxadiazole,

2-(2,6-difluoro-4-(3-(1-(5-(trifluoromethyl)pyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-ethyl-1,3,4- oxadiazole,

2-(2,6-difluoro-4-(3-(1-(5-(trifluoromethyl)pyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,3,4- oxadiazole,

2-(4-(3-(1-(5-chloropyrazin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-methyl-1,3,4-oxadiazole,

2-(4-(3-(1-(5-chloropyrazin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-ethyl-1,3,4-oxadiazole,

2-(4-(3-(1-(5-chloropyrazin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,3,4-oxadiazole,

5-(4-(3-(3,5-difluoro-4-(5-methyl-1,3,4-oxadiazol-2-yl)phenoxy)propyl)piperidin-1-yl)-3-propyl-1 ,2,4-oxadiazole,

5-(4-(3-(3,5-difluoro-4-(5-ethyl-1,3,4-oxadiazol-2-yl)phenoxy)propyl)piperidin-1-yl)-3-propyl-1 ,2,4-oxadiazole,

5-(4-(3-(3,5-difluoro-4-(5-isopropyl-1,3,4-oxadiazol-2-yl)phenoxy)propyl)piperidin-1-yl)-3-propyl-1 ,2,4-oxadiazole,

5-(4-(3-(3,5-difluoro-4-(5-methyl-1,3,4-oxadiazol-2-yl)phenoxy)propyl)piperidin-1-yl)-3-isopropyl-1 ,2,4-oxadiazole,

5-(4-(3-(4-(5-ethyl-1,3,4-oxadiazol-2-yl)-3,5-difluorophenoxy)propyl)piperidin-1-yl)-3-isopropyl-1, 2,4-oxadiazole,

5-(4-(3-(3,5-difluoro-4-(5-isopropyl-1,3,4-oxadiazol-2-yl)phenoxy)propyl)piperidin-1-yl)-3-isopropyl-1 ,2,4-oxadiazole,

5-(4-(3-(3,5-difluoro-4-(5-methyl-1,3,4-oxadiazol-2-yl)phenoxy)propyl)piperidin-1-yl)-3-(2, 2,2-trifluoroethyl)-1,2,4-oxadiazole,

3-(4-(3-(3,5-difluoro-4-(5-isopropyl-1,3,4-oxadiazol-2-yl)phenoxy)propyl)piperidin-1-yl)-5-isopropyl-1 ,2,4-oxadiazole,

2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,3,4-thiadiazole,

2-(2,6-difluoro-4-(3-(1-(5-propylpyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,3,4-thiadiazole,

2-(2,6-difluoro-4-(3-(1-(5-pentylpyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,3,4-thiadiazole,

2-(2,6-difluoro-4-(3-(1-(5-fluoropyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,3,4-thiadiazole,

2-(2,6-difluoro-4-(3-(1-(5-(trifluoromethyl)pyrimidin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,3,4- thiadiazole,

2-(2,6-difluoro-4-(3-(1-(5-(trifluoromethyl)pyridin-2-yl)piperidin-4-yl)propoxy)phenyl)-5-isopropyl-1,3,4- thiadiazole, and

4-ethyl-2-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)thiazole.

In one embodiment, the compound represented by Formula 1 above may be 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl) -5-isopropyl-1,2,4-oxadiazole.

In one embodiment, the pharmaceutical composition may include a compound represented by Formula 1 above; and any one selected from the following: a PPAR agonist, a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist.

The PPAR agonist may be at least one of the following: pioglitazone, lobeglitazone, elafibranor, lanifibranor and saroglitazar. The DPPIV inhibitor may be at least one of the following: sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin and dutogliptin. The ACC inhibitor may be firsocostat. The FXR agonist may be at least one of the following: obeticholic acid, tropifexor, and GW4064 (3-(2,6-dichlorophenyl)-4-(3'-carboxy-2-chlorostilben-4-yl)oxymethyl-5-isopropylisoxazole) . Cenicriviroc could be a dual CCR2/5 antagonist.

For example, the pharmaceutical composition may include a compound represented by Formula 1 above and a PPAR agonist, a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, or a dual CCR2/5 antagonist. The pharmaceutical composition may include 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4- oxadiazole and PPAR agonist. The pharmaceutical composition may include 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4- oxadiazole and DPPIV inhibitor. The pharmaceutical composition may include 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4- oxadiazole and ACC inhibitor. The pharmaceutical composition may include 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4- oxadiazole and FXR agonist. The pharmaceutical composition may include 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4- oxadiazole and dual CCR2/5 antagonist.

The PPAR agonist may be elafibranor. The DPPIV inhibitor may be at least one of the following: evogliptin and sitagliptin. The ACC inhibitor may be firsocostat. An FXR agonist may be obeticholic acid. Cenicriviroc could be a dual CCR2/5 antagonist.

The pharmaceutical composition may include 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4- oxadiazole and elafibranor, evogliptin, sitagliptin, firsocostat, obeticholic acid or cenicriviroc.

The non-alcoholic fatty liver may be selected from the group consisting of the following: non-alcoholic fatty liver, non-alcoholic steatohepatitis, liver fibrosis and cirrhosis.

The pharmaceutical composition may be a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease by inhibiting the deposition of triglycerides in liver tissue.

The pharmaceutical composition may be a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease by inhibiting the occurrence of inflammation in liver tissue.

The pharmaceutical composition may be a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease by inhibiting liver tissue fibrosis.

If the pharmaceutical composition is stated herein to contain A and B, the pharmaceutical composition may be formulated as a composition containing A and B; combinations containing A and B in the form of separate drugs, respectively; or the composition contains a combination. Thus, in the present invention, the term composition can be used interchangeably with the term combination.

The pharmaceutical composition may be one that includes the compound described by formula 1, its pharmaceutically acceptable salts, its optical isomers, its hydrates, its solvates or mixtures thereof; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist, respectively, as a single formulation or contains all as a combination formulation.

The pharmaceutical composition may be a combination of individual drugs or a combination drug.

The pharmaceutical composition may include a first compartment containing a first active ingredient and a second compartment containing a second active ingredient.

The first active ingredient may be a compound of Formula 1, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof, or mixtures thereof. The second active ingredient may include at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist.

In embodiments of the present invention, in a pharmaceutical composition, a first compartment containing a first active ingredient and a second compartment containing a second active ingredient may be co-administered, and thus the pharmaceutical composition may be a co-administration composition. In particular, the pharmaceutical composition may be a combination in which the two ingredients are formulated in separate unit dosage forms. In this case, the first active ingredient and the second active ingredient can be administered together in separate dosage forms.

In one preferred embodiment of the present invention, in a C57BL/6J mouse model in which non-alcoholic fatty liver disease was induced using a special diet, the pharmaceutical composition of one embodiment was found to be effective for the treatment and prevention of non-alcoholic fatty liver disease by histological examination, confirmation of the accumulation of proteins associated with inflammation and fibrosis, experiments to determine the content of AST and ALT, etc.

More specifically, upon administration of the pharmaceutical composition in one embodiment, it was confirmed that the activity score (NAS) of non-alcoholic fatty liver disease (NAFLD) is reduced, the relative content of fat droplets per unit area is reduced, the infiltration of inflammatory cells into liver tissue is reduced, liver tissue fibrosis is suppressed, and the effect of reducing the number of inflammatory and fibrotic biomarkers is enhanced, and the concentration of AST and ALT in the blood is reduced compared to the case when the compound of Formula 1, its pharmaceutically acceptable salts, its optical isomers, its hydrates, its solvates or mixtures thereof are co-administered as GPR119 agonist; or a PPAR agonist, DPPIV inhibitor, ACC inhibitor, FXR agonist, or dual CCR2/5 antagonist, respectively.

In the present invention, a non-limiting example of a pharmaceutically acceptable salt of the compound represented by the above Formula 1 may include salts of inorganic acid such as hydrochloric acid, hydrobromic acid, phosphoric acid or sulfuric acid; organic carboxylic acid salts such as acetic acid, trifluoroacetic acid, citric acid, maleic acid, oxalic acid, succinic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, ascorbic acid or malic acid, or sulfonic acid salts such as methanesulfonic acid or p-toluenesulfonic acid; alkali metal salts such as sodium, potassium or lithium; various acid salts known to form other pharmaceutically acceptable salts; and so on.

The pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease, in one embodiment, can be used in the form of a conventional pharmaceutical preparation. The pharmaceutical preparation may be administered in various oral and parenteral dosage forms, and the dosage form may be variously defined according to the route of administration.

If the pharmaceutical composition of one embodiment is formulated in various oral and parenteral dosage forms, the composition can be prepared by using commonly used excipients such as fillers, diluents, bulking agents, binders, humectants, disintegrants, surfactants, etc. . P.

The solid preparation for oral administration may include tablets, pills, powders, granules, capsules, etc., and these solid preparations can be prepared by mixing the ingredient and at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, gelatin, etc. in a pharmaceutical composition. Also, in addition to simple excipients, lubricants such as magnesium stearate, talc, etc. can be used.

In addition, the oral liquid preparation may include suspending agents, oral liquids, emulsions, syrups, and the like, and may also include various excipients, such as humectants, sweeteners, flavoring agents, preservatives, and the like. .in addition to water and liquid paraffin, which are often used as simple thinners.

The preparation for parenteral administration may include sterile aqueous solutions, non-aqueous diluents, suspensions, emulsions, lyophilizers and suppositories. The non-aqueous solvent and suspending agent used in the present invention may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, an injectable ester such as ethyl oleate, etc. The suppository base used in the present invention may include witepsol, macrogol, tween 61, cocoa butter, laurel oil, glycerin-gelatin, etc.

In addition, a pharmaceutical composition for the prevention or treatment of diabetes or at least one disease selected from related metabolic diseases, when administered, may contain an effective amount ranging from about 1 to about 1000 mg. The amount administered or the amount taken may be administered through a variety of administration doses and techniques, for example, once daily or several times daily by dividing the composition depending on the body weight, age, gender, health status, diet of the subject, time of administration, technique administration, rate of elimination and severity of disease.

In the present invention, non-alcoholic fatty liver disease (NAFLD) may include primary and secondary non-alcoholic fatty liver disease. In particular, in the present invention, non-alcoholic fatty liver disease (NAFLD) may include, but is not limited to, simple steatosis, non-alcoholic steatohepatitis (NASH) and liver fibrosis and cirrhosis caused by the progression of these diseases.

The pharmaceutical composition of one embodiment may include a compound of Formula 1 or an active ingredient having a similar function; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist.

The present invention may also relate to a method of preventing or treating non-alcoholic fatty liver disease, comprising administering to a subject in need of treatment a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formula 1, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof, or mixtures thereof. ; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist.

As used herein, the term “subject in need of treatment” may refer to mammals, including humans, and the term “administration” may refer to providing subjects with predetermined material by a suitable technique. The term "therapeutically effective amount" may be an amount of an active ingredient or pharmaceutical composition that produces biological and medical responses recognized by researchers, veterinarians, physicians or clinicians in animals or humans, which may include the amount necessary to reduce the symptoms of the diseases being treated or violations. Those skilled in the art will appreciate that the therapeutically effective dose and number of administrations of the active ingredient of the present invention may vary depending on the desired effect.

In one embodiment of the route of administration, the pharmaceutical composition proposed in the present invention can be administered by any general route as long as it can reach the target tissue.

Administration may be oral, intraperitoneal, intravenous, intramuscular, subcutaneous, endothelial, nasal, intrapulmonary, rectal, intracavitary, intraperitoneal, and intrathecal, but not limited to.

The pharmaceutical composition of one embodiment can be administered once a day or at least twice a day at certain intervals.

The pharmaceutical composition of one embodiment can be used alone or in combination with surgery, endocrine therapy, drug treatment, and biological response modifier techniques for the prevention and treatment of non-alcoholic fatty liver disease.

One embodiment may also be a nutritional composition for preventing or ameliorating non-alcoholic fatty liver disease, comprising the compound represented by Formula 1 above, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof, or mixtures thereof; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist as an active ingredient.

In one embodiment, the term “relief” may mean all effects in which, as a result of administration of the composition, the disease is alleviated or changed in a favorable manner.

In one embodiment, the term "food" may mean meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, instant pasta, other pasta, chewing gum, dairy products including ice cream, soups, beverages , tea, health drinks, alcoholic drinks, vitamin complexes, functional healthy foods, healthy foods, food with dietary supplements, etc., and includes all types of food in the usual sense of the word.

The term "functional health food" may be the same term as food with dietary supplements (FoSHU), and means a food that results in a significant medical and therapeutic effect, which is processed so that in addition to providing nutrients, it has a biomodulatory function. In this case, "function (functionality)" may mean regulating nutrients or providing a beneficial effect on health when using such physiological effects, etc. in relation to the structure and function of the human body.

“Healthy food” may mean a food that provides an active health maintenance or health enhancing effect compared to regular food and a “nutritional food” can mean a food with a health-promoting nutritional supplement. In some cases, the terms functional health food, health food, and dietary supplement food may be used interchangeably.

The food of the present invention can be prepared according to methods commonly used in the art, and raw materials and ingredients that are commonly added in the art can be added to cook the food during cooking. In particular, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents may be included, and an example of said carbohydrates may include glucose, fructose, maltose, sucrose, oligosaccharide, dextrin, cyclodextrin, xylitol, sorbitol, erythritol, saccharin or synthetic flavoring agents, but is not limited to them only. The food composition of the present invention can be prepared in different dosage forms without imposing restrictions, as long as the dosage form is recognized as food.

The present invention may also relate to a method of preventing or alleviating non-alcoholic fatty liver disease, comprising administering to a subject in need of relief a nutritional composition comprising a compound described by Formula 1 above, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof, or mixtures thereof. ; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist as an active ingredient.

The present invention may also relate to a feed composition for preventing or ameliorating non-alcoholic fatty liver disease, including the compound represented by Formula 1 above, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof or mixtures thereof; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist as an active ingredient.

In the present invention, the term "feed" may mean any natural or artificial diet, single product, etc., or ingredient of said single product that is intended to be consumed or digested by or suitable for livestock. The feed may include feed additives or auxiliary feeds.

There are no particular restrictions on the type of said feed and feed commonly used in the art can be used. A non-limiting example of such feed may include plant foods such as grains, roots, food by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, cake flour, grain processing waste, etc.; animal feed such as proteins, inorganic substances, oils and fats, minerals, unicellular proteins, zooplankton, food products; etc. The feed can be used individually or by mixing at least two of them.

The present invention may also relate to the use of a pharmaceutical composition comprising a compound represented by the above formula 1, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof or mixtures thereof; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist as an active ingredient for the prevention or treatment of non-alcoholic fatty liver disease.

The present invention may also relate to the use of a pharmaceutical composition comprising a compound represented by the above formula 1, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof or mixtures thereof; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist and a dual CCR2/5 antagonist as an active ingredient, for preparing a medicament for the prevention or treatment of non-alcoholic fatty liver disease.

The present invention may also relate to a method of preventing or treating non-alcoholic steatohepatitis, comprising administering to a subject in need of treatment a therapeutically effective amount of a pharmaceutical composition comprising a compound described by the above Formula 1, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof, or mixtures; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist.

The present invention may also relate to a combination of the compound represented by the above formula 1, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof or mixtures thereof; and at least one selected from the following: a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist.

The present invention may also relate to a pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis, comprising a compound described by the above formula 1, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates thereof, solvates thereof or mixtures thereof in combination with at least one selected among the following: DPPIV inhibitor, ACC inhibitor, FXR agonist and dual CCR2/5 antagonist.

In the therapeutic method, food composition, relief method, feed composition, use and combination, the compound represented by the above Formula 1 may preferably be 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin- 4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole.

The materials specified for each pharmaceutical composition, therapeutic method, nutritional composition, relief method, feed composition, use and combination in one embodiment of the present invention are the same for all embodiments unless they conflict with each other.

Beneficial effects of the invention

The pharmaceutical composition of the present invention has an excellent effect on improving lipid metabolism, reducing fat accumulation in liver tissue, and preventing histological damage caused by inflammation and fibrosis of lung tissue, and therefore can be effectively used to prevent or treat non-alcoholic fatty liver disease.

Brief description of drawings

In fig. 1 shows the effect of coadministration on changes in postprandial plasma concentrations of total GLP-1 and active GLP-1 following a single oral administration of the drug to normal mice.

In fig. Figure 2 presents the effect of administration in combination with a DPPIV inhibitor by comparing NAFLD activity before and after multiple doses of the drug in mice induced with nonalcoholic steatohepatitis.

In fig. Figure 3 presents the effect of administration in combination with a PPARα/δ agonist by comparing NAFLD activity before and after multiple doses of the drug in mice induced with nonalcoholic steatohepatitis.

In fig. 4 shows the effectiveness of administration in combination with a DPPIV inhibitor based on quantification of the relative content of lipid droplets in liver tissue after multiple doses of the drug in mice induced with non-alcoholic steatohepatitis.

In fig. Figure 5 shows the effectiveness of administration in combination with a PPARα/δ agonist based on the quantitative determination of the relative content of lipid droplets in liver tissue after administration of several doses of the drug to mice in which non-alcoholic steatohepatitis is caused.

In fig. Figure 6 shows the effectiveness of administration in combination with a DPPIV inhibitor on the relative expression of galectin-3, an indicator of activated macrophages, in liver tissue after multiple doses of the drug in mice with non-alcoholic steatohepatitis.

In fig. Figure 7 shows the effectiveness of the effect of administration in combination with a PPARα/δ agonist on the relative expression of galectin-3, an indicator of activated macrophages in liver tissue after several doses of the drug in mice in which non-alcoholic steatohepatitis is caused.

In fig. Figure 8 shows the effectiveness of administration in combination with a DPPIV inhibitor on the relative expression of α-smooth muscle actin (αSMA), an indicator of activated stellate cells, in liver tissue after multiple doses of the drug in mice induced with non-alcoholic steatohepatitis.

In fig. Figure 9 shows the effectiveness of administration in combination with a PPARα/δ agonist on the relative expression of α-smooth muscle actin (αSMA), an indicator of activated stellate cells, in liver tissue after multiple doses of the drug in mice induced with non-alcoholic steatohepatitis.

In fig. 10 shows the effectiveness of the effect of administration in combination with a DPPIV inhibitor on an indicator of inflammation and fibrosis in plasma after administration of several doses of the drug to mice in which non-alcoholic steatohepatitis is caused.

In fig. 11 shows the effectiveness of the effect of administration in combination with a DPPIV inhibitor on the plasma concentration of the liver damage indicator ALT and AST after administration of several doses of the drug to mice in which non-alcoholic steatohepatitis is caused.

In fig. 12 shows the effect of enhancing the ability to inhibit the secretion of CCL2, TNFα and IL1β, which are inflammatory cytokines, by treatment with the above compound 1 and in combination with a PPARα/δ agonist, an ACC inhibitor, an FXR agonist or a dual CCR2/5 antagonist in the process of human monocyte differentiation into macrophages.

In fig. 13 shows the effect of enhancing the ability to inhibit the expression of fibrotic proteins by treatment with the above compound 1 and in combination with a PPARα/δ agonist, an ACC inhibitor or an FXR agonist in the activation of primary human hepatic stellate cells by TGFβ.

Mode of implementation of the invention

The features and advantages of the present invention, as well as methods for providing them, are apparent with reference to the exemplary embodiments described in detail below in the present invention. However, the present invention is not limited to the typical embodiments disclosed below in the present invention, but can be implemented in various different forms. Below in the present invention, the following exemplary embodiments are provided for a better understanding of the present invention and are presented only to fully illustrate the scope of the present invention to those skilled in the art, and thus, the present invention is defined only by the scope of its claims.

Non-alcoholic steatohepatitis is not a disease caused by a single factor, and therefore it may be difficult to expect a superior therapeutic effect with a single drug. Accordingly, a pharmaceutical composition of one embodiment of the present invention containing a plurality of active ingredients can improve the effect of preventing or treating non-alcoholic steatohepatitis when administered in combination with additional drugs with different mechanisms of action.

Drugs currently being developed as therapeutics for non-alcoholic steatohepatitis are roughly divided into i) drugs to reduce fat accumulation in liver tissue by improving metabolism, ii) drugs to reduce oxidative stress in liver tissue or suppress inflammatory responses by directly regulating them, iii) drugs to suppress hepatocyte apoptosis and iv) drugs to suppress the accumulation of extracellular fibrotic proteins.

In particular, metabolic diseases such as obesity and diabetes are characterized by high prevalence and have etiological characteristics in which metabolic imbalance causes fat accumulation in liver tissue, and thus antidiabetic drugs with different mechanisms of action previously developed to improve metabolism have been confirmed , are effective in animal models of nonalcoholic steatohepatitis or are under clinical investigation.

The pharmaceutical composition of one embodiment of the present invention may include a GPR119 agonist and at least one agent selected from the following: an activated peroxisome proliferator receptor (PPAR) agonist, a dipeptidyl peptidase IV (DPPIV) inhibitor, an acetyl-Co-A carboxylase (ACC) inhibitor, farnesoid X receptor (FXR) agonist and dual C-C chemokine receptor 2/5 (CCR2/5) antagonist. Thus, the pharmaceutical composition can exhibit an excellent effect of treating or preventing non-alcoholic steatohepatitis by complementary effects of the active ingredients described above. More specifically, the pharmaceutical composition of one embodiment may result in a synergistic effect in the treatment or prevention of non-alcoholic steatohepatitis as opposed to administering each of the above-described active ingredients separately.

As used herein, the term "at least one" may include all combinations formed from a plurality of configurations. For example, "at least one of A, B and C" may include "A, B or C", "two selected from A, B and C" and all combinations selected from "A, B and C".

GPR119 may be one of the G protein-coupled receptors (GPCRs) to which Gs binds to activate signaling through cAMP. GPR119 is highly expressed in pancreatic beta cells and small intestinal L cells and is also found in the liver and acts as a regulator of fatty acid biosynthesis in the liver. In addition, when GP119 is activated, all of the following can be regulated: fatty liver, inflammation and fibrosis in non-alcoholic steatohepatitis by suppressing the differentiation and activation of inflammatory cells and activation of hepatic stellate cells.

Mechanistically, when GPR119 in the small intestine is activated by dietary fatty acid derivatives, GPR119 can enhance the secretion of glucagon-like peptide-1 (GLP-1). Secreted GLP-1 is inactivated in the blood by an enzyme called dipeptidyl peptidase IV (DPPIV) within minutes.

GLP-1 may have pharmacological effects to improve metabolism and, as a GLP-1-like peptide, may clinically reduce fatty liver in patients with nonalcoholic steatohepatitis.

According to the mechanism described above, by administering a GPR119 agonist (eg, a compound of Formula 1 above) in combination with a DPPIV inhibitor designed as a hypoglycemic agent, the duration of biological activity of active GLP-1 in the blood can be prolonged. Accordingly, it may be possible to demonstrate the effect of a GPR119 agonist in the treatment or prevention of non-alcoholic steatohepatitis and, simultaneously, the pharmacological effect of GLP-1 in improving metabolism.

In other words, the pharmaceutical composition of one embodiment comprising a GPR119 agonist and a DPPIV inhibitor may complement the metabolic-enhancing effects of GLP-1 in addition to the medicinal effects of the GPR119 agonist itself in ameliorating non-alcoholic steatohepatitis. Thus, the pharmaceutical composition can be considered to have a synergistic effect in the treatment or prevention of non-alcoholic steatohepatitis.

The GPR119 agonist may be a GPR119 ligand, and more preferably may be a compound of Formula 1 described above.

The pharmaceutical composition of one embodiment may include different combinations of active ingredients in addition to the combination of a GPR119 agonist and a DDPIV inhibitor. For example, the pharmaceutical composition of one embodiment may include the above-described PPAR agonist or ACC inhibitor as ingredients that enhance the effectiveness of treatment or prevention of non-alcoholic steatohepatitis by improving metabolism in combination with a GPR119 agonist, in addition to a DPPIV inhibitor, which is the ingredient leading to the effect improve metabolism together with the GPR119 agonist. Alternatively, the pharmaceutical composition may include an FXR agonist or a dual CCR2/5 antagonist, which in combination with a GPR119 agonist may further reduce inflammation and fibrosis in patients with non-alcoholic steatohepatitis.

More specifically, the PPAR agonist may be at least one of the following: pioglitazone, lobeglitazone, elafibranor, lanifibranor and saroglitazar; the DPPIV inhibitor may be at least one of the following: sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin and dutogliptin; and the ACC inhibitor may be firsocostat. The FXR agonist may be at least one of the following: obeticholic acid, tropifexor, and GW4064 (3-(2,6-dichlorophenyl)-4-(3'-carboxy-2-chlorostilben-4-yl)oxymethyl-5-isopropylisoxazole) . Cenicriviroc could be a dual CCR2/5 antagonist.

The pharmaceutical composition of one embodiment may include a GPR119 agonist; and at least one selected from the following: a PPAR agonist, a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist described above.

The pharmaceutical composition of one embodiment may include a compound represented by the above Formula 1 described above; and at least one selected from the following: a PPAR agonist, a DPPIV inhibitor, an ACC inhibitor, an FXR agonist, and a dual CCR2/5 antagonist.

The pharmaceutical composition of one embodiment may include 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2 ,4-oxadiazole; and at least one of the following: pioglitazone, lobeglitazone, elafibranor, lanifibranor, saroglitazar, sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gozogliptin and dutogliptin, firsoco stat, obeticholic acid, tropifexor , GW4064 and tsenikrivirok.

The pharmaceutical composition of one embodiment may include a combination of ingredients. Thus, when administered, the pharmaceutical composition of one embodiment can provide a much greater effect in treating or preventing non-alcoholic steatohepatitis than when the individual ingredients are administered separately.

Below in the present invention, the pharmaceutical composition in one embodiment of the present invention is described in detail with reference to examples and comparative examples.

<Example 1> Confirmation of the effectiveness of a combination of GPR119 ligand and drugs with other mechanisms of action in a mouse model in which steatohepatitis is induced using a special diet

To confirm the effectiveness of the pharmaceutical composition of one embodiment of the present invention in the treatment of non-alcoholic fatty liver disease, the following experiment was conducted.

Confirmation of the mechanism of action in normal mice

Male ICR mice, 7 weeks old, were administered a single oral dose of 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin-4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl -1,2,4-oxadiazole (hereinafter referred to as “compound 1”) in the compound of Formula 1 and sitagliptin as a DPPIV inhibitor at a dose of 3 mg/kg and 10 mg/kg, respectively, separately or in combination. Glucose solution was administered orally 30 min after administration. After oral glucose administration, blood was drawn 10 min later and the concentration of total GLP-1 (ALPCO, 43-GPTHU-E01) and the concentration of active GLP-1 in plasma (ALPCO, 43-GP1HU-E01) were quantified using an ELISA kit and the results are shown in fig. 1.

As confirmed in FIG. 1, when a GPR119 agonist and a DPPIV inhibitor were used in combination, the total GLP-1 concentration and the peak plasma concentration of the active form were increased compared to when Compound 1 was administered alone. This means that when drugs with both mechanisms of action are used in combination, the secretion of GLP-1 in the small intestine is increased and the biological half-life of GLP-1 from the blood is increased.

Study of a mouse model of non-alcoholic steatohepatitis

Male C57BL/6J mice, 6 weeks old, were fed a special high-fat, high-fructose, high-cholesterol diet (Research Diet, D09100301) for at least 27 weeks and were thus induced to develop nonalcoholic steatohepatitis. Compound 1 was then administered alone or in combination so that the daily dose reached 100 mg/kg/day. As a DPPIV inhibitor, evogliptin 200 mg/kg/day or sitagliptin 150 mg/kg/day was administered alone or in combination. As a PPAR agonist, elafibranor was administered alone or in combination to achieve a daily dose of 30 mg/kg/day. For the single or combination administration method, Compound 1, a DPPIV inhibitor, and/or a PPAR agonist described above was formulated as a drug-feed mixture and given for 8 to 12 weeks.

Histological examination

For histological examination, liver tissue was extracted by biopsy and autopsy in a mouse model of induced non-alcoholic steatohepatitis before and after drug administration and fixed in 10% formaldehyde to form a paraffin block, and tissue sections with a thickness of 2 μm were obtained. Hematoxylin and eosin (HE) staining was then performed using an automatic stainer (Autostainer XL, Leica) to confirm inflammatory cell infiltration. The content of lipid droplets deposited on the stained liver tissues, the degree of inflammatory cell infiltration and the degree of cell necrosis were expressed as NAS activity indices of NAFLD and the results are shown in Fig. 2 and 3.

To quantitatively compare fat accumulation in liver tissue, image analysis of HE-stained tissue was performed to calculate the relative content of lipid droplets per unit area and thereby determine the drug efficacy of co-administration. The results are shown in Fig. 4 and 5.

To compare the degree of inflammatory cell infiltration in liver tissue, the expression level of galectin-3, an indicator of increased expression of activated macrophages, was determined using immunostaining. The galectin-3-stained antibody content per unit area was then determined by image analysis to determine drug efficacy in reducing inflammation due to co-administration. The results are shown in Fig. 6 and 7.

To determine efficacy in fibrosis, α-smooth muscle actin (αSMA) expression in liver section tissue was determined by immunostaining to correlate the content of activated stellate cells. The content of αSMA-stained antibodies per unit area was then determined by image analysis. The results are shown in Fig. 8 and 9.

Confirmation of accumulation of inflammation- and fibrosis-related proteins

To study the effect on the accumulation of inflammation-related proteins (CCL2, CXCl10, CXCl2, IL2 and TNFα) and fibrosis-related proteins (TIMP1) in plasma, the concentration of proteins CCL2, CXCl10, CXCl2, IL2, TNFα (MSD, K15255D-1) and TIMP1 (R&D Systems, MTM100) in plasma in a mouse model was quantified using a commercially available ELISA kit. The results are shown in Fig. 10.

Determination of AST and ALT content in blood

Mice model of induced nonalcoholic steatohepatitis were necropsied and plasma was collected for quantification of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) using an automated blood analyzer (Konelab 20i). The results are shown in Fig. eleven.

As confirmed in FIG. 2, the NAS score of mice treated with Compound 1 alone was significantly improved compared with the pre-treatment value, but the effect of evogliptin, a DPPIV inhibitor, administered alone was not significant. However, when both drugs were administered in combination, NAS increased significantly.

As shown in FIG. 3, when compound 1 or elafibranor (a PPAR agonist) was administered alone, the NAS score was significantly improved compared to the pre-treatment value and when both drugs were administered in combination, the NAS score was further significantly increased.

As shown in FIG. 4, the size of lipid droplets in liver tissues was decreased in mice administered Compound 1 alone, and thus the relative area of lipid droplets was significantly decreased. When the DPPIV inhibitor evogliptin was administered alone, there was a trend toward a decrease in the relative area of lipid droplets. First of all, when compound 1 was administered in combination with a DPPIV inhibitor, the relative area of lipid droplets in liver tissue was further reduced, confirming the increased drug efficacy in reducing liver fatty infiltration when using the combination of both drugs.

As confirmed in FIG. 5, according to the quantitative determination of the degree of fat deposition in the liver tissue, the fat content in the liver tissue was significantly reduced when compound 1 or PPAR agonist was administered alone. As a result of administration of Compound 1 in combination with a PPAR agonist, lipid droplet area was further reduced, demonstrating an improvement of up to 94% compared to the control group (NASH Co.). This demonstrates an increase in drug efficacy in reducing fatty liver when using a combination of both drugs.

As shown in FIG. 6, the expression of galectin-3, an indicator of activated macrophages in liver tissue, was significantly reduced in mice treated with compound 1 or evogliptin, a DPPIV inhibitor, alone, and when both drugs are used in combination, the effect of reducing galectin-3 expression is further enhanced. This shows that when both drugs are administered in combination, the effect of reducing inflammatory cell infiltration in liver tissue is enhanced.

As shown in FIG. 7, the expression of galectin-3, an indicator of activated macrophages in liver tissue, was significantly decreased in mice treated with compound 1 alone. When elafibranor, a PPAR agonist, was administered alone, the effect of significantly reducing galectin-3 expression was not observed, but when both drugs were used in combination, the effect of reducing galectin-3 expression was confirmed to be further enhanced. This indicates an enhanced effect of reducing the infiltration of inflammatory cells in liver tissue when using a combination of both drugs.

As shown in FIG. 8, the expression of αSMA, which is an indicator of hepatic stellate cell activation causing liver tissue fibrosis, was significantly decreased in mice treated with compound 1 or the DPPIV inhibitor evogliptin alone. This shows that the effect of inhibiting αSMA expression is further enhanced when both drugs are administered in combination. This shows that the effect of suppressing liver tissue fibrosis is enhanced when using a combination of both drugs.

As shown in FIG. 9, the expression of αSMA, an indicator of hepatic stellate cell activation causing liver tissue fibrosis, was significantly decreased in mice administered Compound 1 alone, whereas no significant inhibition effect on αSMA expression was observed when elafibranor, a PPAR agonist, was administered alone. However, it was confirmed that the inhibition of αSMA expression was further enhanced when both drugs were administered in combination. This indicates an enhanced effect of suppressing liver tissue fibrosis when both drugs are administered together.

In fig. 10 shows that there is an effect on blood concentrations of inflammatory and fibrotic biomarkers when Compound 1 is administered alone or in combination with sitagliptin, a DPPIV inhibitor. Plasma concentrations of CCL2, CXCl10, and CXCl2, chemokines that induce inflammatory cell infiltration, were increased in NASH mice, and their plasma concentrations were significantly decreased when compound 1 was administered alone. When compound 1 was used in combination with a DPPIV inhibitor, the drug's chemokine reduction efficacy was further increased. There was a trend toward a decrease in the inflammatory cytokines IL2 and TNFα when Compound 1 was administered alone, but was significantly reduced when combined with a DPPIV inhibitor. TIMP1, an indicator of fibrosis, was also significantly increased in NASH mice, but was significantly decreased when compound 1 was administered alone, and the decreasing effect was enhanced when combined with a DPPIV inhibitor. Accordingly, it is confirmed that the effect of reducing the amount of histologically promoting inflammatory and fibrotic biomarkers in plasma is enhanced when Compound 1 and a DPPIV inhibitor are used in combination.

In fig. 11 depicts the effect of a pharmaceutical composition of one embodiment on plasma ALT and AST as an indicator of liver damage. When compound 1 or the DPPIV inhibitor evogliptin was repeatedly administered alone, plasma concentrations of ALT and AST were significantly decreased. When Compound 1 was administered in combination with a DPPIV inhibitor, ALT and AST concentrations were further reduced, significantly different from when both drugs were administered alone. This shows that the effect of preventing hepatocyte damage caused by non-alcoholic steatohepatitis is enhanced when Compound 1 and a DPPIV inhibitor are administered in combination.

The above results indicate that administration of a pharmaceutical composition of one embodiment of the present invention in combination with a GPR119 agonist and a metabolic disease attenuator such as a DPPIV inhibitor or a PPAR agonist reduces fatty liver and suppresses liver damage, inflammation and fibrosis, thereby providing influence on non-alcoholic fatty infiltration of the liver and exhibits a more effective therapeutic effect.

<Example 2> Confirmation of the effectiveness of the combination of the GPR119 ligand and drugs with other mechanisms of action on human macrophages

Confirmation of accumulation of inflammatory cytokine proteins secreted in the environment

To study in vitro the possibility of enhancing the effect of reducing inflammation among the effects of reducing non-alcoholic fatty liver when treated with the GPR119 ligand in combination with drugs with other mechanisms of action, the content of inflammatory cytokines in the medium was quantified to confirm the effect of the combination of the GPR119 ligand and drugs with other mechanisms effects on the differentiation of human monocytes.

Human monocyte line (THP-1, ATCC ^® TIB-202™) cells were treated with 50 ng/ml phorbol 12-myristate-13-acetate (PMA) for 48 hours and they differentiated into macrophages when treated with Compound 1 in combination with elafibranor as a PPAR agonist, firsocostat as an ACC inhibitor, obeticholic acid as an FXR agonist, or cenicriviroc as a dual CCR2/5 antagonist, then the medium was changed to serum-free medium and then treated with lipopolysaccharides (LPS; 0.5 ng/ml) for 4 h, after which the content of interleukin-1β (IL-1β; R&D Systems, DY201), tumor necrosis factor α (TNFα; R&D Systems, DY210), and CC motif chemokine ligand 2 (CCL2; R&D Systems, DY279), which are inflammatory cytokines secreted in the medium were quantified using a commercially available ELISA kit to examine the effect on monocyte differentiation, and the results are shown in FIG. 12.

As confirmed in FIG. 12, when treated with Compound 1 of the present invention, during the differentiation process of monocytes into macrophages, it was confirmed that the secretion of IL-1β, TNFα and CCL2 in accordance with the activation of macrophages was decreased in a concentration-dependent manner. When treated with a PPAR agonist, an ACC inhibitor, an FXR agonist, or a dual CCR2/5 antagonist in combination, the ability of Compound 1 to inhibit the secretion of inflammatory cytokines was enhanced.

The above results indicate that Compound 1 of the present invention suppresses the differentiation of immune cells and suppresses the secretion of inflammatory cytokines caused by innate immunity, thereby inhibiting the progression of non-alcoholic fatty liver disease, and thus, this effect can be enhanced by treatment in combination with a PPAR agonist, an ACC inhibitor, an FXR agonist, or a dual CCR2/5 antagonist.

<Example 3> Confirmation of the effectiveness of the combination of GPR119 ligand and drugs with other mechanisms of action in human hepatic stellate cells

Confirmation of the expression level of fibrotic protein expressed in cells

To investigate in vitro the potential for enhanced fibrosis-inhibiting effects among the effects of reducing non-alcoholic fatty liver disease when co-treated with GPR119 ligand and drugs with other mechanisms of action, the concentration of fibrotic proteins expressed in primary human hepatic stellate cells was quantified.

Primary human liver stellate cells (ScienCell, #5300) were stabilized for 24 hours, then grown in serum-free medium for 24 hours, and then treated with Compound 1 in combination with elafibranor elafibranor as a PPAR agonist, firsocostat as an ACC inhibitor, or obeticholic acid as an FXR agonist along with 5 ng/ml recombinant tumor growth factor β1 (TGFβ1), after which the resulting cells were harvested after 18 h and the amount of procollagen 1a1 in total cellular proteins was quantified using a commercially available ELISA kit (R&D Systems, DY6220) , and the amount of total cellular proteins was quantified and corrected by the BCA method, and the results of the concentration response curves for each drug and the concentration (IC ₃₀ ) representing 30% inhibitory activity are shown in FIG. 13.

As can be confirmed from the data in Fig. 13, when treated with Compound 1 alone, the expression of procollagen 1a1 protein induced by TGFβ 1 was decreased in a concentration-dependent manner and the 30% inhibitory concentration was found to be 0.55 μM. A PPAR agonist, an ACC inhibitor, or an FXR agonist resulted in a relatively weak response. Treat with 0.3 μM of the above compound 1 along with a PPAR agonist, ACC inhibitor, or FXR agonist at a concentration that is predicted to result in 30% inhibition when treated together, and the proportion of concentration at which both drugs result in 30% inhibition is calculated. inhibitory activity when treated together compared with treatment with the drug alone, so that their sum was the combination index (CI) (Chou TC & Talalay P., Adv Enzyme Regul, 1984:22:27-55). When treated with Compound 1 in combination with a PPAR agonist or an ACC inhibitor, the CI value was found to be 0.7 and 0.6, which is less than 1, confirming the synergistic effect. In addition, the CI value found to be 1 when treated in combination with an FXR agonist confirms the additive effect.

The above results show that Compound 1 of the present invention suppresses the activation of hepatic stellate cells to suppress the expression of collagen, which is a fibrous protein, and thus suppresses the progression of non-alcoholic fatty liver disease and thus the effect can be enhanced by treatment in combination with a PPAR agonist, an ACC inhibitor, or an FXR agonist.

Claims (28)

1. A pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease, containing: the compound represented by the following chemical formula 1, its pharmaceutically acceptable salts, its optical isomers, its hydrates, its solvates or mixtures thereof; And at least one selected from pioglitazone, lobeglitazone, elafibranor, lanifibranor, saroglitazar, sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, dutogliptin ina, firsocostat, obeticholic acid, tropifexor, 3-(2,6-dichlorophenyl)-4-(3'-carboxy-2-chlorostilben-4-yl)oxymethyl-5-isopropylisoxazole and cenicriviroc: [Formula 1] where in the above formula 1 A is an oxadiazole group, wherein said A is substituted with a C1-C6 linear or branched alkyl group; B is a pyrimidine group, wherein said B is substituted with a C1-C6 linear or branched alkyl group; And X stands for F. 2. Pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, where the above A means or , And – R ₂ and R ₃ each independently represent a C1-C6 linear or branched alkyl group. 3. Pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, where the above B means , And R ₇ represents a C1-C6 linear or branched alkyl group. 4. Pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 1, where the compound described by the above formula 1 is 3-(4-(3-(1-(5-ethylpyrimidin-2-yl)piperidin- 4-yl)propoxy)-2,6-difluorophenyl)-5-isopropyl-1,2,4-oxadiazole. 5. The pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver disease according to claim 4, where the pharmaceutical composition contains at least one of elafibranor, evogliptin, sitagliptin, firsocostat, obeticholic acid and cenicriviroc. 6. Use of a pharmaceutical composition containing: the compound described by chemical formula 1 according to claim 1, its pharmaceutically acceptable salts, its optical isomers, its hydrates, its solvates or mixtures thereof; and at least one selected from pioglitazone, lobeglitazone, elafibranor, lanifibranor, saroglitazar, sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, dutogly ptina, firsocostat, obeticholic acid, tropifexor , 3-(2,6-dichlorophenyl)-4-(3'-carboxy-2-chlorostilben-4-yl)oxymethyl-5-isopropylisoxazole and cenicriviroc as the active ingredient for the prevention or treatment of non-alcoholic fatty liver disease. 7. Use of a pharmaceutical composition containing: the compound described by chemical formula 1 according to claim 1, its pharmaceutically acceptable salts, its optical isomers, its hydrates, its solvates or mixtures thereof; and at least one selected from pioglitazone, lobeglitazone, elafibranor, lanifibranor, saroglitazar, sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, dutogly ptina, firsocostat, obeticholic acid, tropifexor , 3-(2,6-dichlorophenyl)-4-(3'-carboxy-2-chlorostilben-4-yl)oxymethyl-5-isopropylisoxazole and cenicriviroc as an active ingredient for the preparation of a medicinal product for the prevention or treatment of non-alcoholic fatty liver disease . 8. A method for preventing or treating non-alcoholic fatty liver disease, comprising: administering a therapeutically effective amount of a pharmaceutical composition comprising a compound described by chemical formula 1 according to claim 1, its pharmaceutically acceptable salts, its optical isomers, its hydrates, its solvates or mixtures thereof; and at least one selected from pioglitazone, lobeglitazone, elafibranor, lanifibranor, saroglitazar, sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, dutogly ptina, firsocostat, obeticholic acid, tropifexor , 3-(2,6-dichlorophenyl)-4-(3'-carboxy-2-chlorostilben-4-yl)oxymethyl-5-isopropylisoxazole and cenicriviroc as the active ingredient in a subject in need of treatment. 9. Use according to claim 6, wherein the non-alcoholic fatty liver disease is selected from the group consisting of simple fatty liver disease, non-alcoholic steatohepatitis, liver fibrosis and cirrhosis. 10. Use according to claim 6, where the pharmaceutical composition inhibits the occurrence of inflammation in liver tissue and fibrosis of liver tissue. 11. Use according to claim 7, wherein the non-alcoholic fatty liver disease is selected from the group consisting of simple fatty liver disease, non-alcoholic steatohepatitis, liver fibrosis and cirrhosis. 12. Use according to claim 7, where the pharmaceutical composition inhibits the occurrence of inflammation in liver tissue and fibrosis of liver tissue. 13. The method of claim 8, wherein the non-alcoholic fatty liver disease is selected from the group consisting of simple fatty liver disease, non-alcoholic steatohepatitis, liver fibrosis and cirrhosis. 14. The method according to claim 8, where the pharmaceutical composition inhibits the occurrence of inflammation in the liver tissue and fibrosis of the liver tissue.