Novel 2-arylthiazole derivatives or salts thereof, a method for producing the same, and a pharmaceutical composition containing the same

Abstract

The present invention provides novel 2-arylthiazole derivatives or pharmaceutically acceptable salts thereof, which have a specific carboxamide moiety, such as a substituted aminoalkyl-carboxamide moiety, an N-containing heterocycle-alkyl-carboxamide moiety, or an N-containing heterocycle-carboxamide moiety, a method for preparing the same, and pharmaceutical compositions containing the same.

Description

Technical Field

[0001] The present invention relates to novel 2-arylthiazole derivatives or salts thereof, a method for producing the same, and a pharmaceutical composition containing the same. More specifically, the present invention relates to novel 2-arylthiazole derivatives or pharmaceutically acceptable salts thereof having a specific carboxamide moiety, such as a substituted aminoalkyl-carboxamide moiety, an N-containing heterocyclic-alkyl-carboxamide moiety, or an N-containing heterocyclic-carboxamide moiety, a method for producing the same, and a pharmaceutical composition containing the same. The 2-arylthiazole derivative or pharmaceutically acceptable salt thereof of the present invention has excellent autophagy-inducing activity.

Background Art

[0002] Autophagy, also called autophagocytosis, is a natural intracellular regulatory mechanism that degrades unnecessary components or components that have malfunctioned. Autophagy enables the orderly degradation and reuse of cell components. In the process of autophagy, consumable cytoplasmic components are sequestered inside double-membrane vesicles known as autophagosomes and separated from the remaining cell components. Next, the autophagosome fuses with available lysosomes, and the contents of the vesicle are finally degraded and reused. There are generally three forms of autophagy: macroautophagy, microautophagy, and chaperone-mediated autophagy (CMA). When a disease develops, autophagy is considered to promote cell survival as an adaptive response to stress, but it may also promote cell death and pathological conditions. In extreme cases such as starvation, the energy level of cells is maintained by the degradation of cell components, promoting cell survival.

[0003] On the one hand, when autophagy decreases, various diseases may occur due to the accumulation of misfolded proteins, etc. For example, it has been reported that neurodegenerative diseases such as Huntington's disease (HD), Parkinson's disease (PD), Alzheimer's disease (AD), prion disease, multiple sclerosis, amyotrophic lateral sclerosis (Lou Gehrig's disease), etc. can be treated by inducing autophagy (for example, Korean Patent No. 10-1731908). Also, it has been reported that liver diseases such as liver fibrosis, cirrhosis, hepatitis, fatty liver disease, etc. can be treated by inducing autophagy (for example, Korean Patent Publication No. 10-2017-0022790). Furthermore, it has been reported that metabolic diseases such as diabetes, hyperlipidemia, obesity, inflammation, etc. can be treated by inducing autophagy (for example, Korean Patent Publication No. 10-2018-0007307). Additionally, it has been reported that the excessive immune response associated with sepsis can be suppressed by inducing autophagy (for example, Korean Patent Publication No. 10-2012-0131401).

[0004] Therefore, substances that induce autophagy are expected to be usefully applied to the prevention or treatment of various autophagy-related diseases such as neurodegenerative diseases, liver diseases, metabolic diseases, sepsis, etc.

Summary of the Invention

Problems to be Solved by the Invention

[0005] The inventors have found that novel 2-arylthiazole derivatives or pharmaceutically acceptable salts thereof having a specific carboxamide moiety, for example, a substituted aminoalkyl-carboxamide moiety, an N-containing heterocyclic-alkyl-carboxamide moiety, or an N-containing heterocyclic-carboxamide moiety, have excellent autophagy-inducing activity and can thus be usefully applied to the prevention or treatment of various autophagy-related diseases.

[0006] Therefore, the present invention provides the above-mentioned 2-arylthiazole derivatives or pharmaceutically acceptable salts thereof, a method for producing the same, a pharmaceutical composition containing the same, and uses thereof.

Means for Solving the Problem

[0007] According to one aspect of the present invention, a novel 2-arylthiazole derivative or a pharmaceutically acceptable salt thereof is provided.

[0008] According to another aspect of the present invention, a method for producing the 2-arylthiazole derivative or a pharmaceutically acceptable salt thereof is provided.

[0009] According to still another aspect of the present invention, a pharmaceutical composition containing the 2-arylthiazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient is provided.

[0010] According to still another aspect of the present invention, a method for treating an autophagy-related disease, which includes administering the 2-arylthiazole derivative or a pharmaceutically acceptable salt thereof, is provided.

[0011] According to still another aspect of the present invention, the use of the 2-arylthiazole derivative or a pharmaceutically acceptable salt thereof for producing a prophylactic or therapeutic agent for an autophagy-related disease is provided.

Advantages of the Invention

[0012] The compound of the present invention, namely, a novel 2-arylthiazole derivative or a pharmaceutically acceptable salt thereof having a specific carboxamide moiety, such as a substituted aminoalkyl-carboxamide moiety, an N-containing heterocyclic-alkyl-carboxamide moiety, or an N-containing heterocyclic-carboxamide moiety, has excellent autophagy-inducing activity. Therefore, the compound of the present invention or a pharmaceutically acceptable salt thereof can be usefully applied to the prevention or treatment of various autophagy-related diseases such as neurodegenerative diseases, liver diseases, metabolic diseases, and sepsis.

Brief Description of the Drawings

[0013]

Figure 1

Figure 2

Figure 3

Figure 4

Mode for Carrying Out the Invention

[0014] In the present specification, "alkyl" refers to a linear or branched aliphatic hydrocarbon group. For example, C 1 ~C 6 alkyl means a linear or branched aliphatic hydrocarbon having 1 to 6 carbon atoms such as methyl, ethyl, propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, and isopentyl.

[0015] "Hydroxy" refers to an -OH group. "Alkoxy" refers to a group formed by substituting the hydrogen atom of a hydroxyl group with an alkyl. For example, C 1 ~C 6 alkoxy includes methoxy, ethoxy, propoxy, n-butoxy, n-pentyloxy, isopropoxy, isobutoxy, sec-butoxy, tert-butoxy, neopentyloxy, and isopentyloxy.

[0016] "Amino" refers to the -NH 2 group. "Alkylamino" refers to an amino group substituted with mono- or dialkyl. For example, C 1-6 alkylamino contains an amino group substituted with mono- or di-C 1-6 alkyl.

[0017] The present invention provides a compound having excellent autophagy-inducing activity or a salt thereof, that is, a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0018] In the compound of the present invention or a pharmaceutically acceptable salt thereof, R 1 is preferably hydrogen; an isobutyl group; or a diethylaminoethyl group, and R 2 is preferably a methyl group.

[0019] In the compound of the present invention or a pharmaceutically acceptable salt thereof, A is a group selected from the group consisting of the groups of Chemical Formulas 1a to 1d. In one embodiment of the present invention, A may be a group of Chemical Formula 1a or a group of Chemical Formula 1b. The compound of the present invention in which A is a group of Chemical Formula 1a or a group of Chemical Formula 1b has the structures of the following Chemical Formulas 11a and 11b.

Chemical Formula

Chemical Formula

[0020] In the compounds of Chemical Formulas 11a and 11b, R 1 , R 2 , R3 , R 4 , R 5 , and R 6 is the same as that defined above.

[0021] In the compounds of Formulas 11a and 11b, R 3 and R 4 may preferably combine with the nitrogen atom to which they are attached to form piperidine, morpholine, or pyrrolidine. More preferably, said piperidine, morpholine, or pyrrolidine may be substituted with C 1 -C 4 alkyl.

[0022] More preferably, the compound of the present invention or a pharmaceutically acceptable salt thereof is N-(2-(diethylamino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(diisopropylamino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(dimethylamino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(3-(diethylamino)propyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(4-methylhomopiperazino)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(4-(1-methyl)piperidinyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(4-morpholino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(1-Pyrrolidino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(1-Piperidino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((1-(Dimethylamino)cyclopentyl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((1-(Dimethylamino)cyclohexyl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(3-(1-Benzyl)pyrrolidinyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(2-(1-Methyl)pyrrolidino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((S)-2-(1-Ethyl)pyrrolidinomethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(4-Morpholinoamino)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(1-Piperidinoamino)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diisopropylamino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diethylamino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Dimethylamino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(4-Morpholino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((S)-2-(1-Ethyl)pyrrolidinomethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diisopropylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diethylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Dimethylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(4-Morpholino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((S)-2-(1-Ethyl)pyrrolidinomethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; and N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride may be one or more selected from the group consisting of.

[0023] Particularly preferably, the compound of the present invention or a pharmaceutically acceptable salt thereof is N-(2-(Diisopropylamino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diisopropylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(4-Morpholino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; and N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride which may be one or more selected from the group consisting of.

[0024] The compound of Formula 1 may be in the form of a pharmaceutically acceptable salt, for example, an acid addition salt. The acid addition salts may be in the form of salts derived from inorganic acids or organic acids such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acetate, lactate, citrate, tartrate, succinate, maleate, malonate, oxalate, fumarate, gluconate, saccharinate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate, etc., but are not limited thereto. The acid addition salts may be prepared by reacting the compound of Formula 1 with an inorganic acid or an organic acid in a conventional solvent such as water, alcohol, tetrahydrofuran, acetone, or a mixture thereof.

[0025] The compound of Formula 1 according to the present invention or a pharmaceutically acceptable salt thereof may be in an anhydrous form, a hydrated form or a solvated form. Further, the compound of Formula 1 according to the present invention or a pharmaceutically acceptable salt thereof may be in an amorphous form or a crystalline form. The amorphous form or the crystalline form may be a hydrated form or a solvated form. The hydrate or solvate may contain a stoichiometric or non-stoichiometric amount of water or an organic solvent with respect to the compound of Formula 1.

[0026] The compound of Formula 1 or a pharmaceutically acceptable salt thereof may have a substituent containing an asymmetric carbon, and thus may be in the form of a racemic mixture (RS), or in the form of an optical isomer such as an (R) isomer or an (S) isomer. Therefore, unless otherwise specified, the compound of Formula 1 or a pharmaceutically acceptable salt thereof includes both a racemic mixture (RS) and optical isomers such as an (R) isomer or an (S) isomer.

[0027] The present invention also includes within its scope a method for producing the compound of Formula 1 or a pharmaceutically acceptable salt thereof. For example, the compound of Formula 1 or a pharmaceutically acceptable salt thereof may be produced by acylating the compound of Formula 2 with an N-containing compound to prepare the compound of Formula 1, as shown in Reaction Scheme 1 below, or by acylating the compound of Formula 3 with an N-containing compound to prepare the compound of Formula 4; and alkylating the compound of Formula 4 with R 1 X to prepare the compound of Formula 1.

Chemical Formula

[0028] In Reaction Scheme 1, R 1 , R 2 and A are the same as those defined above, and X is a halogen.

[0029] In one embodiment, the present invention provides a method of reacting a compound of Formula 2 with NH2 -(CH 2 ) n -NR 3 R 4 、NH 2 -CH 2 -CR 5 R 6 -NR 3 R 4 、Cy, and NH 2 -Cy selected from the group consisting of N-containing compounds (wherein n, R 3 , R 4 , R 5 , and R 6 are the same as those defined above; Cy is a nitrogen-containing heterocyclic group selected from the group consisting of homopiperazine, piperidine, morpholine, and pyrrolidine, and the nitrogen-containing heterocyclic group is optionally substituted with C 1 ~C 4 alkyl or benzyl; NH 2 -Cy is a nitrogen-containing heterocyclic amine selected from the group consisting of homopiperazinylamine, piperidinylamine, morpholinylamine, and pyrrolidinylamine, and the nitrogen-containing heterocyclic amine is optionally substituted with C 1 ~C 4 alkyl or benzyl) to acylate, to provide a method for producing a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof:

Chemical Formula

Chemical Formula

[0030] In another embodiment, the present invention provides a compound of Chemical Formula 3 with NH 2 -(CH 2 ) n -NR 3 R 4 , NH 2 -CH 2 -CR 5 R6 -NR 3 R 4 、 Cy, and NH 2 An N-containing compound selected from the group consisting of -Cy (wherein n, R 3 、R 4 、R 5 、and R 6 are the same as those defined above; Cy is a nitrogen-containing heterocyclic group selected from the group consisting of homopiperazine, piperidine, morpholine, and pyrrolidine, and the nitrogen-containing heterocyclic group is optionally substituted with C 1 ~C 4 alkyl or benzyl; NH 2 -Cy is a nitrogen-containing heterocyclic amine selected from the group consisting of homopiperazinylamine, piperidinylamine, morpholinylamine, and pyrrolidinylamine, and the nitrogen-containing heterocyclic amine is optionally substituted with C 1 ~C 4 alkyl or benzyl) is used for acylation to prepare a compound of Chemical Formula 4; and a step of alkylating the compound of Chemical Formula 4 with a compound of Chemical Formula 5 to prepare a compound of Chemical Formula 1, provided is a method for producing a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof:

Chemical Structure

Chemical Structure

Chemical Structure

[0031] A compound of Chemical Formula 2, a compound of Chemical Formula 3, and an N-containing compound (i.e., NH 2 -(CH 2 ) n NR 3 R 4, NH 2 -CH 2 -CR 5 R 6 -NR 3 R 4 , Cy, and NH 2 -Cy) are known compounds and are commercially available. Acylation may be carried out using an acylating agent such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), dicyclohexylcarbodiimide (DCC), 1,1'-carbonyldiimidazole (CDI), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). Further, acylation may be carried out by reacting a compound of Chemical Formula 2 or a compound of Chemical Formula 3 with thionyl chloride, oxalyl chloride, phosphorus chloride, etc. to produce an acyl chloride, and then reacting with an N-containing compound. Acylation may be carried out in a conventional organic solvent, for example, a solvent such as dichloromethane, dimethylformamide.

[0032] Alkylation may be carried out in the presence of an alkali metal base such as sodium hydroxide or sodium carbonate. The reaction between the compound of Chemical Formula 4 and R 1 X may be carried out at a molar ratio of 1:1 to 1:5, preferably about 1:1. Alkylation may be carried out in water, C 1 ~C 4 alcohol, acetone, tetrahydrofuran, toluene, or a mixture thereof.

[0033] The 2-arylthiazole derivative of the present invention, that is, the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof, has excellent autophagy-inducing activity. Therefore, the compound of Chemical Formula 1 of the present invention or a pharmaceutically acceptable salt thereof can be usefully applied to the prevention or treatment of various autophagy-related diseases such as neurodegenerative diseases, liver diseases, metabolic diseases, and sepsis.

[0034] Therefore, the present invention includes within its scope a pharmaceutical composition for inducing autophagy containing, as an active ingredient, a therapeutically effective amount of the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[0035] Autophagy-related diseases include, but are not limited to, various diseases that can be prevented, ameliorated, or treated by inducing autophagy. For example, the pharmaceutical composition of the present invention is a neurodegenerative disease selected from the group consisting of Huntington's disease, Parkinson's disease, Alzheimer's disease, prion disease, multiple sclerosis, and Lou Gehrig's disease; a liver disease selected from the group consisting of liver fibrosis, cirrhosis, hepatitis, and fatty liver disease; a metabolic disease selected from the group consisting of diabetes, hyperlipidemia, obesity, and inflammation; or a pharmaceutical composition for preventing or treating sepsis.

[0036] The pharmaceutical composition of the present invention may contain pharmaceutically acceptable carriers such as diluents, disintegrants, sweeteners, lubricants, flavoring agents, etc. The pharmaceutical composition of the present invention may be formulated into oral dosage forms such as tablets, capsules, powders, granules, suspensions, emulsions, syrups, etc.; or parenteral dosage forms such as external solutions, external suspensions, external emulsions, gels (e.g., ointments), inhalants, sprays, injections, etc. according to conventional methods. These dosage forms may be in various forms, for example, they may be dosage forms for single administration or dosage forms for multiple administrations.

[0037] The pharmaceutical composition of the present invention may contain, for example, diluents such as lactose and corn starch; lubricants such as magnesium stearate; emulsifying agents; suspending agents; stabilizing agents; and / or isotonic agents, etc. If necessary, the composition of the present invention may further contain sweeteners and / or flavoring agents.

[0038] The composition of the present invention may be administered orally or parenterally by routes such as inhalation, intravenous, intraperitoneal, subcutaneous, intracerebroventricular, rectal, and topical administration routes. Therefore, the composition of the present invention can be formulated into various forms such as tablets, capsules, aqueous solutions, suspensions, etc. In the case of tablets for oral administration, carriers such as lactose and corn starch, and lubricants such as magnesium stearate are conventionally used. In the case of capsules for oral administration, lactose and / or dried corn starch can be used as diluents. When an aqueous suspension for oral administration is required, an emulsifier and / or a suspending agent may be blended with the active ingredient. If necessary, specific sweeteners and / or flavoring agents may be used. In the case of intramuscular, intraperitoneal, subcutaneous, or intravenous administration, usually, a sterile solution of the active ingredient needs to be prepared, and a buffer is added to the solution to appropriately adjust the pH. In the case of intravenous administration, it is necessary to adjust the total concentration of the solute to make the formulation isotonic. The composition of the present invention may be in the form of an aqueous solution containing a pharmaceutically acceptable carrier such as physiological saline at pH 7.4. This solution may be introduced into the bloodstream in the muscle of the patient by local bolus injection.

[0039] The 2-arylthiazole derivative of the present invention, that is, the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof, may be administered to a patient in a therapeutically effective amount of about 0.0001 mg / kg to about 100 mg / kg per day, preferably about 0.001 mg / kg to about 100 mg / kg per day. The administration may be carried out once a day or several times a day via the oral route or the parenteral route. Needless to say, this dosage may be changed according to the age, condition, weight, sensitivity, degree of the disease, administration route, administration period, etc. of the patient. The pharmaceutical composition according to the present invention may contain the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof in an amount of 0.001 to 99% by weight, preferably 0.01 to 60% by weight, depending on the administration method.

[0040] The present invention includes within its scope a method for inducing autophagy in a mammal in need of autophagy induction, the method comprising administering to the mammal a therapeutically effective amount of a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof. For example, the present invention includes a neurodegenerative disease selected from the group consisting of Huntington's disease, Parkinson's disease, Alzheimer's disease, prion disease, multiple sclerosis, and Lou Gehrig's disease; a liver disease selected from the group consisting of liver fibrosis, cirrhosis, hepatitis, and fatty liver disease; a metabolic disease selected from the group consisting of diabetes, hyperlipidemia, obesity, and inflammation; or a method for preventing or treating sepsis.

[0041] The present invention also includes within its scope the use of a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for inducing autophagy in a mammal in need of autophagy induction. For example, the present invention includes the use of a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for a neurodegenerative disease selected from the group consisting of Huntington's disease, Parkinson's disease, Alzheimer's disease, prion disease, multiple sclerosis, and Lou Gehrig's disease; a liver disease selected from the group consisting of liver fibrosis, cirrhosis, hepatitis, and fatty liver disease; a metabolic disease selected from the group consisting of diabetes, hyperlipidemia, obesity, and inflammation; or a medicament for preventing or treating sepsis.

[0042] The following examples and test examples are provided for the purpose of illustrating the present invention and do not limit the scope of the present invention in any way.

[0043] The analysis of the compounds produced in the following examples was carried out as follows. Nuclear magnetic resonance (NMR) spectrum analysis was performed using a Bruker spectrometer (400 MHz), and chemical shifts were analyzed in units of ppm. Column chromatography was carried out using silica gel (Merck, 70 - 230 mesh). Unless otherwise specified, all starting materials were purchased as commercial products and used without further purification. All reactants and chromatographic fractions were analyzed by thin - layer chromatography (TLC) using silica gel plates at 250 nm and visualized by ultraviolet light or iodine (I 2 ) staining. The products and intermediates were purified by flash chromatography or reverse - phase HPLC.

[0044] Example 1: Preparation of N-(2-(diethylamino)ethyl)-2-(3 - cyano - 4 - isobutoxyphenyl)-4 - methylthiazole - 5 - carboxamide hydrochloride

Chemical formula

[0045] Example 2: Preparation of N-(2-(diisopropylamino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0046] Example 3: Preparation of N-(2-(dimethylamino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical Structure

[0047] Example 4: Preparation of N-(3-(diethylamino)propyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0048] Example 5: Preparation of N-(4-methylhomopiperazino)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0049] Example 6: Preparation of N-(4-(1-methyl)piperidinyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0050] Example 7: Preparation of N-(2-(4-Morpholino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide Hydrochloride

Chemical Structure

[0051] Example 8: Preparation of N-(2-(1-Pyrrolidino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide Hydrochloride

Chemical Structure

[0052] Example 9: Preparation of N-(2-(1-Piperidino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide Hydrochloride

Chemical Structure

[0053] Example 10: Preparation of N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride [Chemical formula] The title compound was prepared according to the same procedure as in Example 1, using 4-(aminomethyl)-N,N-dimethyltetrahydro-2H-pyran-4-amine instead of 2-(diethylamino)ethylamine. (Yield: 67.9%) TLC R f = 0.44 in 10% MeOH in Chloroform 1 H NMR (400 MHz, DMSO-d 6) δ 10.31 (t, 1H, J = 4.8 Hz), 8.67 (t, 1H, J = 6.4 Hz), 8.26 (d, 1H, J = 2.4 Hz), 8.20 (dd, 1H, J = 2.4 Hz, 8.8 Hz), 7.40 (d, 1H, J = 8.8 Hz), 4.02 (d, 2H, J = 6.4 Hz), 3.95 - 3.85 (m, 2H), 3.83 (d, 2H, J = 6.4 Hz), 3.65 - 3.55 (m, 2H), 2.80 (s, 3H), 2.79 (s, 3H), 2.64 (s, 3H), 2.15 - 2.05 (m, 1H), 1.95 - 1.85 (m, 4H), 1.03 (d, 6H, J = 6.4 Hz)

[0054] Example 11: Preparation of N-((1-(dimethylamino)cyclopentyl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0055] Example 12: Preparation of N-((1-(dimethylamino)cyclohexyl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0056] Example 13: Preparation of N-(3-(1-benzyl)pyrrolidinyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0057] Example 14: Preparation of N-(2-(2-(1-methyl)pyrrolidino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0058] Example 15: Preparation of N-((S)-2-(1-Ethyl)pyrrolidinomethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide Hydrochloride

Chemical Structure

[0059] Example 16: Preparation of N-(4-morpholinoamino)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical Structure

[0060] Example 17: Preparation of N-(1-piperidineamino)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical Structure

[0061] Example 18: Preparation of N-(2-(Diisopropylamino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide Hydrochloride [Chemical formula] A mixture of 2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxylic acid (3.00 g), hydroxybenzotriazole (1.71 g), dicyclohexylcarbodiimide (2.62 g) and dimethylformamide (30 ml) was stirred for 10 minutes. N-Methylmorpholine (1.28 g) and 2-(diisopropylamino)ethylamine (1.83 g) were added to the mixture, and after raising the temperature to 50 °C, the mixture was stirred for 3 hours. The reaction mixture was cooled to 0 - 5 °C, and the resulting solid was removed by filtration. The filtrate was concentrated under reduced pressure. The obtained oily concentrate was dissolved in 0.5 N hydrochloric acid solution (60 ml) and purified water (60 ml), and then washed with ethyl acetate (60 ml × 2 times). The aqueous layer was adjusted to pH 7 - 8 with sodium hydroxide (1.2 g) and extracted with dichloromethane (60 ml × 2 times). The combined extracts were dried over anhydrous sodium sulfate and then concentrated in vacuo to obtain N-(2-(diisopropylamino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide. This compound was dissolved in methanol (10 ml), and then an ethereal solution of 1 N hydrochloric acid (20 ml) was added. After heating the mixture to 40 °C, acetone (30 ml) was added. The mixture was stirred for 1 hour and then filtered. The obtained solid was washed with acetone (10 ml) and then dried at 50 - 55 °C to obtain 4.17 g of the title compound. (Yield: 85.5%) TLC R f = 0.20 in 20% MeOH in Chloroform 1 H NMR (400 MHz, MeOH-d 4 ) δ 8.18 (d, 1H, J = 2.4 Hz), 8.09 (dd, 1H, J = 2.4 Hz, 8.8 Hz), 7.11 (d, 1H, J = 8.8 Hz), 3.90 ~ 3.82 (m, 2H), 3.74 (t, 2H, J = 6.8 Hz), 3.38 (t, 2H, J = 6.8 Hz), 2.73 (s, 3H), 1.46 (d, 12H, J = 6.8 Hz)

[0062] Example 19: Preparation of N-(2-(diethylamino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chem.

[0063] Example 20: Preparation of N-(2-(dimethylamino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chem.

[0064] Example 21: Preparation of N-(2-(4-Morpholino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide Hydrochloride

Chem.

[0065] Example 22: Preparation of N-((S)-2-(1-Ethyl)pyrrolidinomethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide Hydrochloride

Chem.

[0066] Example 23: Preparation of N-(2-(diisopropylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0067] Example 24: Preparation of N-(2-(diethylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0068] Example 25: Preparation of N-(2-(dimethylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0069] Example 26: Preparation of N-(2-(4-morpholino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride

Chemical formula

[0070] Example 27: Preparation of N-((S)-2-(1-Ethyl)pyrrolidinomethyl)-2-(4-(2-Diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide Hydrochloride [Chemical formula] Using the N-((S)-2-(1-Ethyl)pyrrolidinomethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride prepared in Example 22, the title compound was prepared according to the same procedure as in Example 23. (Yield: 28.8%) TLC R f = 0.08 in 20% MeOH in Chloroform 1 H NMR (400MHz, DMSO-d 6) δ 10.70 (s, 1H), 10.66 (s, 1H), 8.80 (t, 1H, J = 5.6 Hz), 8.31 (d, 1H, J = 2.4 Hz), 8.25 (dd, 1H, J = 2.4 Hz, 8.8 Hz), 7.45 (d, 1H, J = 8.8 Hz), 4.65 (t, 2H, J = 4.4 Hz), 3.80 - 3.50 (m, 6H), 3.45 - 3.35 (m, 5H), 3.15 - 3.05 (m, 2H), 2.67 (s, 3H), 2.20 - 2.10 (m, 1H), 2.05 - 1.80 (m, 3H), 1.35 - 1.25 (m, 9H)

[0071] Example 28: Preparation of N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride [Chemical formula] A mixture of 2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxylic acid (5.00 g), hydroxybenzotriazole (2.86 g), dicyclohexylcarbodiimide (4.36 g) and dimethylformamide (45 ml) was stirred for 10 minutes. N-Methylmorpholine (2.14 g) and 4-(aminomethyl)-N,N-dimethyltetrahydro-2H-pyran-4-amine (3.50 g) were added to the mixture, and after raising the temperature to 50 °C, the mixture was stirred for 4 hours. The reaction mixture was cooled to 0 - 5 °C, and the resulting solid was removed by filtration. The filtrate was concentrated under reduced pressure to obtain N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide. This compound was dissolved in 2N hydrochloric acid solution (25 ml) and purified water (100 ml), washed with ethyl acetate (100 ml), and then filtered. The resulting filtrate was stirred for 1 hour and then filtered. The resulting solid was washed with cold purified water (10 ml), and then acetone (50 ml) was added. The mixture was stirred for 30 minutes and then filtered. The resulting solid was washed with acetone (20 ml) and dried at 50 - 55 °C to obtain 7.10 g of the title compound. (Yield: 84.6%) TLC R f = 0.48 in 20% MeOH in Chloroform 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.98 (s, 1H), 10.48 (s, 1H), 8.67 (t, 1H, J = 6.4 Hz), 8.15 (d, 1H, J = 2.4 Hz), 8.06 (dd, 1H, J = 6.4 Hz, 8.8 Hz), 7.21 (d, 1H, J = 8.8 Hz), 3.92 ~ 3.85 (m, 2H), 3.83 (d, 2H, J = 6.4 Hz), 3.68 ~ 3.58 (m, 2H), 2.79 (s, 6H), 2.63 (s, 3H), 2.00 ~ 1.88 (m, 4H)

[0072] Example 29: Preparation of N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride [Chemical formula] Using the N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride prepared in Example 28, the title compound was produced according to the same procedure as in Example 23. (Yield: 74.0%) TLC R f = 0.38 in 20% MeOH in Chloroform 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.45 ~ 10.35 (m, 2H), 8.72 (t, 1H, J = 6.4 Hz), 8.33 (d, 1H, J = 2.4 Hz), 8.27 (dd, 1H, J = 6.4 Hz, 8.8 Hz), 7.45 (d, 1H, J = 8.8 Hz), 4.64 (t, 2H, J = 4.4 Hz), 3.98 ~ 3.85 (m, 2H), 3.84 (d, 2H, J = 6.4 Hz), 3.70 ~ 3.60 (m, 4H), 3.35 ~ 3.25 (m, 4H), 2.80 (d, 6H, J = 4.8Hz), 2.65 (s, 3H), 1.95 ~ 1.88 (m, 4H), 1.30 (t, 6H, J = 7.2 Hz)

[0073] Test Example 1: Autophagy induction activity By using an autophagy detection kit (DAPGreen Autophagy detection, Dojindo, D676-10) according to the manufacturer's instructions, the autophagy induction activity of the compound of the present invention was measured in Hela cells (Korean Cell Line Bank). Specifically, Hela cells (8×10 4Cells were seeded into each well of a 96-well plate together with FluroBrite DMEM containing 10% fetal bovine serum (FBS), and then incubated overnight at 37°C in a CO 2 incubator to stabilize the cells. A DAPGreen (Cat. D676-10, Dojindo) solution (0.1 mM DMSO solution) was diluted 1 / 1000 with medium (FluroBrite DMEM containing 10% FBS) to prepare a medium containing 0.1 μM DAPGreen (DAPGreen / FBS-containing medium). After removing the supernatant from each well where the cells had adhered in a monolayer, each well was washed with FluroBrite DMEM. The DAPGreen / FBS-containing medium (100 μl) prepared above was added to each well, and the plate was 2 incubated in a CO incubator for 30 minutes. Fluorescence values (EX: 450 nm / EM: 535 nm) were measured at the start and 30 minutes after the start of DAPGreen treatment, respectively. The difference value was taken as the basal absorption value of the cells.

[0074] Similar to the measurement of the basal absorption value, after removing the supernatant from each well where the cells had adhered in a monolayer, each well was washed with FluroBrite DMEM. The compounds of the examples, the reference substance (2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxylic acid), and the positive reference substance (rapamycin) were each dissolved in DMSO. Each solution was diluted to concentrations of 500 nM and 1000 nM with DAPGreen / FBS-containing medium (100 μl) and added to the cells. Three wells were tested for each concentration (n = 3). After incubation at 37°C for 4 hours, fluorescence values (EX: 450 nm / EM: 535 nm) were measured. After subtracting the basal absorption value from the fluorescence value of each well, each obtained value was corrected (normalized) based on the average absorption value of the DMSO-treated group.

[0075] The relative fluorescence values of the test substances (1000 nm and 500 nm) and the control substance (2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxylic acid: 1000 nm and 500 nm) were calculated based on the fluorescence values obtained by treatment with the positive control substance (rapamycin: 1000 nm and 500 nm). The results are shown in Table 1 below. In Table 1, when the autophagy-inducing activity of the test substance prepared in the example is equivalent to that of the positive control substance, the relative fluorescence value is 1. When the test substance prepared in the example shows higher autophagy-inducing activity than the positive control substance, the relative fluorescence value exceeds 1, and when the test substance prepared in the example shows lower autophagy-inducing activity than the positive control substance, the relative fluorescence value is less than 1.

Table 1

[0076] From the results shown in Table 1 above, when incubated for 4 hours after treatment with each test substance, the compounds according to the present invention showed autophagy-inducing activity equivalent to or higher than that of the positive control substance (i.e., rapamycin), and also showed significantly superior autophagy-inducing activity than 2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxylic acid having a carboxylic acid moiety. Therefore, since the compounds according to the present invention show excellent autophagy-inducing activity, they can be usefully applied to the prevention and treatment of various autophagy-related diseases such as neurodegenerative diseases, liver diseases, metabolic diseases, and sepsis.

[0077] Test Example 2: Analysis of autophagosome formation CYTO-ID TMThe autophagy lysosomal pathway (ALP) activation by the compounds of the present invention was verified by measuring the degree of autophagosome formation in neurons using an Autophagy Detection Kit (ENZ-51031-K200, Enzo Life Sciences, Inc.). Neurons (i.e., primary mouse cortical neuron cells) were isolated from mice using an enzymatic digestion method. Specifically, cortical tissue from 16-day-old C57BL6 mouse embryos was incubated with 20 U / ml of papain (Worthington Biochemical Corporation, LK003176) and 0.005% of DNase I (Worthington Biochemical Corporation, LK003170) at 37°C for 30 minutes to isolate primary mouse cortical neurons.

[0078] Primary mouse cortical neurons (8×10 4 cells) and medium (2 mM L-glutamine (Gibco, 25030-081), N2 supplement (Gibco, 17502-048), B27 supplement (Gibco, 17504-044), and 50 μg / ml penicillin-streptomycin (Gibco, 15140-122)) were added to each well of a 96-well plate and incubated overnight in a 37°C, CO 2 incubator. The compounds of Example 10 (5 μM), the compounds of Example 29 (5 μM), and rapamycin (10 μM) were added to the cells respectively and incubated in a 37°C, CO 2 incubator for 24 hours. After washing each well with 1× Assay Buffer (Enzo Life Science, ENZ-51031-K200), 0.2 μl of Cyto-ID autophagy detection dye and 0.1 μl of Hoechst 33342 were added to the cells and incubated at 37°C, CO 2It was incubated in an incubator for 30 minutes. After each well was washed with 1× Assay Buffer (Enzo Life Science, ENZ-51031-K200), the fluorescence values of Cyto-ID (EX: 480 nm, EM: 530 nm) and Hoechst 33342 (EX: 340 nm, EM: 480 nm) were measured with a fluorometer respectively. Three wells were tested for each group. After the Cyto-ID fluorescence value of each well was divided by the Hoechst 33342 fluorescence value for correction (normalization), the relative intensity was calculated with the untreated control group set as 1. The results are shown in Table 2 below.

Table 2

[0079] From the results in Table 2 above, it can be seen that the compounds of Example 10 and 29 can significantly increase autophagosome formation even at a concentration of 1 / 2 compared with the positive control substance (i.e., rapamycin). Therefore, since the compounds according to the present invention exhibit excellent autophagy-inducing activity, they can be usefully applied to the prevention and treatment of various autophagy-related diseases such as neurodegenerative diseases, liver diseases, metabolic diseases, and sepsis.

[0080] Test Example 3: Autophagy Flux Analysis N2a cells (ATCC, CCL-131, mouse neuroblastoma cell line) (3×10 4 cells) and medium (DMEM (Dulbecco’s modified Eagle’s medium, Gibco, 11995-065) supplemented with 5% heat-inactivated fetal bovine serum (FBS, Gibco, 16000-044) and 50 μg / ml penicillin-streptomycin (Gibco, 15140-122)) were added to each well of a 6-well plate and incubated overnight in an incubator at 37 °C, CO 2 The compound of Example 29 (5 μM) and rapamycin (10 μM) were added respectively and incubated at 37 °C, CO 2It was incubated in an incubator for 24 hours. For each group, blocks for electron microscopy were prepared in 3 wells, and pre-fixation, post-fixation, dehydration, substitution, and embedding were performed by the flat embedding method. Ultra-thin sections with a thickness of 70 nm were prepared, and after heavy metal staining (uranyl acetate, lead citrate) was performed on each section, it was photographed by the mosaic imaging method that images the entire cell. The results of electron microscopy imaging are shown in Figure 1. Also, the results obtained by measuring the number of total autophagosomes (autophagosome vesicles), autophagosomes, and autolysosomes per cell for each treatment group are shown in Table 3 below. In Table 3 below, the control group means the untreated control group. [Table 3]

[0081] From the results of Table 3 above, it can be seen that the compound of Example 29 significantly increases autophagosome vesicle formation, particularly autolysosome formation, even at a concentration of 1 / 2 compared to the positive control substance (i.e., rapamycin). Therefore, since the compound according to the present invention exhibits excellent autophagy-inducing activity, it can be usefully applied to the prevention and treatment of various autophagy-related diseases such as neurodegenerative diseases, liver diseases, metabolic diseases, and sepsis.

[0082] Test Example 4: Evaluation of liver function improvement activity by oral administration in a liver injury model (1) Male Sprague-Dawley rats induced with liver injury by dimethylnitrosamine (DMN) were orally administered the compound according to the present invention for 3 weeks to evaluate the liver function improving activity. Specifically, 7-week-old male Sprague-Dawley rats (Orient Bio, Korea) were acclimated to the laboratory environment at room temperature for 7 days. Overall symptoms were observed, and only healthy rats were used in the experiment. The rats were divided into 4 groups (n = 5 per group), namely, a normal control group, a group administered only DMN, and groups administered both the compound of the present invention (the compound of Example 2 or Example 23) and DMN. DMN was dissolved in purified water and intraperitoneally administered at a dose of 10 mg / kg for 3 consecutive days per week (3 times a week for 4 weeks). Blood samples were collected 3 days after the completion of liver injury induction in the first week, and serum ALT (alanine transaminase) values and serum AST (aspartate transaminase) values were measured to confirm liver injury. The compound of the present invention was administered for 3 weeks, that is, from the 4th day after the completion of liver injury induction in the first week to during the DMN administration period. Each compound of the present invention was dissolved in purified water and orally administered once a day at a dose of 25 mg / kg for 3 weeks using an oral sonde. Blood samples were collected on day 0 (3 days after the completion of liver injury induction in the first week) and on days 7, 14, and 21 after the administration of the test substance. The collected blood was injected into a vacutainer tube containing a blood clotting accelerator and left at room temperature for about 20 minutes to clot each blood sample. After centrifugation for 10 minutes, blood biochemical tests were performed using the obtained serum. The outline of the experimental method is shown in Figure 2. The results obtained from the measurements were compared between groups by parametric multiple comparison or non-parametric multiple comparison using SPSS. When P < 0.05, it was judged to be statistically significant.

[0083] The serum ALT values and serum AST values obtained by performing the blood biochemical test as described above are shown in Tables 4 and 5 below. [Table 4] [Table 5]

[0084] As can be seen from the results shown in Tables 4 and 5, the serum ALT value in the DMN-administered group after 3 weeks became about 3 times, and the serum AST value became about 2 times. However, in the group administered both the compound of the present invention and DMN, the ALT values 3 weeks after administration were 101.04 and 117.02 U / L (i.e., a 35% and 25% decrease compared to the DMN-administered group); the AST values 3 weeks after administration were 203.44 and 231.54 U / L (i.e., a 32% and 23% decrease compared to the DMN-administered group). Therefore, it can be confirmed that the compound of the present invention has excellent inhibitory activity against liver fibrosis.

[0085] Test Example 5: Evaluation of liver function improving activity by oral administration in a liver injury model (2) Male Sprague-Dawley rats induced with liver injury by dimethylnitrosamine (DMN) were orally administered the compound according to the present invention for 4 weeks to evaluate the liver function improving activity. Specifically, 7-week-old male Sprague-Dawley rats (Orient Bio, Korea) were acclimated to the laboratory environment at room temperature for 7 days. The overall symptoms were observed, and only healthy rats were used in the experiment. The rats were divided into 5 groups (n = 10 for each group), namely, a normal control group, a group administered only DMN, and groups administered both the compound of the present invention (the compound of Example 23, 26, or 29) and DMN. DMN was dissolved in purified water and intraperitoneally administered at a dose of 10 mg / kg for 3 consecutive days per week (3 times a week for 4 weeks). After completion of the induction of liver injury in the 4th week, blood samples were collected, and serum ALT (alanine transaminase) values and serum AST (aspartate transaminase) values were measured to confirm liver injury. The compound of the present invention was administered for 4 weeks starting from the first day after completion of the induction of liver injury in the 4th week. The compound of the present invention was dissolved in purified water and orally administered once a day at a dose of 25 mg / kg for 4 weeks using an oral sonde. Blood samples were collected on day 0 (1 day after completion of the induction of liver injury in the 4th week) and on days 7, 14, 21, and 28 after administration of the test substance. The collected blood was injected into a vacutainer tube containing a coagulation accelerator and left at room temperature for about 20 minutes to coagulate each blood sample. After centrifugation for 10 minutes, blood biochemical tests were performed using the obtained serum. Also, 24 hours after the last administration, autopsy was performed, and the excised liver was fixed to prepare tissue specimens. Slides were prepared from the tissue specimens and stained with hematoxylin and eosin (H&E) to microscopically observe the damage to the liver tissue and the infiltration of inflammatory cells in the liver tissue. Also, Masson's trichrome staining was performed to microscopically observe the deposition of collagen fibers in the liver tissue. The outline of the experimental method is shown in Figure 3. The results obtained from the measurement were compared between groups by parametric multiple comparison or non-parametric multiple comparison using SPSS. When P < 0.05, it was determined to be statistically significant. The serum ALT values and serum AST values obtained by performing the blood biochemical test as described above are shown in Tables 6 and 7 below.

Table 6

Table 7

[0086] As can be seen from the results shown in Table 6 and Table 7, the serum ALT value in the DMN-administered group after 4 weeks became about 4 to 5 times, and the serum AST value became about 3 to 4 times. However, in the group administered both the compound of the present invention and DMN, the ALT value 4 weeks after administration was 52.93 to 59.12 U / L (that is, a 9.9 to 19% decrease compared to the DMN-administered group); the AST value 4 weeks after administration was 114.45 to 125.59 U / L (that is, a 21 to 28% decrease compared to the DMN-administered group). In particular, the compound-administered group of Example 26 showed an AST value equivalent to that of the normal control group 4 weeks after administration. Therefore, it can be confirmed that the compound of the present invention has excellent therapeutic activity against liver fibrosis. Also, as described above, autopsy was performed on the normal control group, the DMN-administered group (4 weeks), and the group administered both the compound of the present invention (the compound of Example 23 or 26) and DMN to prepare liver tissue specimens, and the results obtained by performing hematoxylin-eosin (H&E) staining and Masson trichrome staining are shown in FIG. 4. From the results shown in FIG. 4, the degree of damage to the liver tissue and the degree of infiltration of inflammatory cells in the liver tissue were measured. Also, from the results shown in FIG. 4, the degree of deposition of collagen fibers in the liver tissue was measured. These results are shown in Table 8 below.

Table 8

[0087] As can be seen from the results of H&E staining (FIG. 4) and Table 8, compared with the normal control group, the damage to the liver tissue and the infiltration of inflammatory cells in the liver tissue in the DMN-administered group after 4 weeks increased by about 17 times and about 5 times, respectively. However, in the group administered both the compound of the present invention (the compound of Example 23 or 26) and DMN, the damage to the liver tissue and the infiltration of inflammatory cells in the liver tissue were 259.14 to 319.14 cells and 90.00 to 101.43 cells / mm per 1000 cells, respectively 2This was the case (i.e., there were improvements of about 43 - 54% and about 46 - 52% respectively compared to the DMN administration group). Also, as can be seen from the results of Masson's trichrome staining (Figure 4) and Table 8, the deposition of collagen fibers in the liver tissue of the DMN administration group after 4 weeks increased by about 12 - fold compared to the normal control group. However, in the group administered both the compound of the present invention (the compound of Example 23 or 26) and DMN, the deposition of collagen fibers in the liver tissue was 10.75 - 12.44% / mm 2 This was the case (i.e., there was a decrease of about 57 - 63% compared to the DMN administration group). Therefore, it can be confirmed that the compounds of the present invention (the compounds of Example 23 and 26) have excellent inhibitory activity against liver fibrosis.

[0088] Test Example 6: Analysis of inhibitory activity against beta - amyloid - induced neuronal cell death The inhibitory activity of the compounds of the present invention against beta - amyloid - induced neuronal cell death, which is known as a major cytotoxic substance in neurodegenerative diseases, was analyzed in N2a cells (ATCC, CCL - 131, mouse neuroblastoma cell line) and primary mouse cortical neuron cells isolated in the same manner as in Test Example 2. The medium used for N2a cells was the same as that used in Test Example 3, and the medium used for primary mouse cortical neuron cells was the same as that used in Test Example 2. Specifically, N2a cells (3×10 4 cells) and primary mouse cortical neuron cells (8×10 4 cells) were added to each well of a 96 - well plate together with their respective media and incubated overnight in a 37°C, CO 2 incubator. The cells were treated with the compound of Example 10 and the compound of Example 29 at concentrations of 5 μM and 500 nM respectively, and then treated with 20 μM of beta - amyloid 30 minutes later, and cultured in a 37°C, CO 2 incubator for 72 hours. After the cells were treated with 10 μL of CCK - 8 reagent (Enzo Life Science, ALX - 850 - 039 - KI02), they were incubated at 37°C, CO 2It was incubated in an incubator for 2 hours. Each group was tested in three wells, and each absorbance was measured at 450 nm. The absorbance value of the untreated control group was set to 100% cell viability. The cell viability of each group was calculated from the respective absorbance values. The results are shown in Table 9 below. In Table 9, the control group indicates the group treated with only 20 μM of beta-amyloid.

Table 9

[0089] From the results in Table 9, it can be confirmed that both the compound of Example 10 and the compound of Example 29 significantly suppress the cytotoxicity caused by beta-amyloid treatment. Therefore, the compounds according to the present invention can be usefully applied to the prevention and treatment of various neurodegenerative diseases related to beta-amyloid.

[0090] Test Example 7: Analysis of the inhibitory activity against mitochondrial damage by rotenone The inhibitory activity of the compounds of the present invention against mitochondrial damage caused by rotenone, which is known as a major cytotoxic substance in neurodegenerative diseases, was analyzed in N2a cells (ATCC, CCL-131, mouse neuroblastoma cell line). Specifically, N2a cells (3×10 4 cells) and medium (DMEM (Dulbecco’s modified Eagle’s medium, Gibco, 11995-065) supplemented with 5% heat-inactivated fetal bovine serum (FBS, Gibco, 16000-044) and 50 μg / ml penicillin-streptomycin (Gibco, 15140-122)) were added to each well of an XF24 well culture plate and incubated overnight in a 37 °C, CO 2 incubator. The cells were treated with the compound of Example 10 (5 μM), and after 30 minutes, treated with 10 μM of rotenone, and then incubated at 37 °C, CO 2It was incubated in an incubator for 24 hours. After treatment with the XF Cell Mito Stress Test Kit (Seahorse Bioscience, 103015-100) according to the manufacturer's instructions, the level of mitochondrial respiration was measured using the XF24 Extracellular Flux Analyzer (Seahorse Bioscience). Four wells were tested for each group, and basal respiration, ATP production, maximal respiration, and spare respiratory capacity were measured. The level of each mitochondrial respiration was corrected (normalized) by the protein quantification value. The basal respiration of the untreated control group was set to 100%, and the remaining values were shown. These results are shown in Table 10 below. In Table 10, the control group represents the group treated with only 10 μM of rotenone.

Table 10

[0091] From the results in Table 10, it can be confirmed that the compound of Example 10 restores mitochondrial damage caused by rotenone treatment to a statistically significant level. Therefore, the compound according to the present invention can be usefully applied to the prevention and treatment of various neurodegenerative diseases associated with mitochondrial damage.

Claims

Claim 1 A compound of Formula 1 or a pharmaceutically acceptable salt thereof: 【Chemical 1】 Wherein, R １ is hydrogen; C １ to C ４ alkyl group; or mono- or di-C １ to C ５ alkylamino-substituted C １ to C ４ alkyl group, R ２ is a C １ -C ４ alkyl group, A is a group selected from the group consisting of the groups of the following Formulas 1a - 1d, [Chemical 2] [Chemical Formula 3] 【Chemical Formula 4】 [Chemical Formula 5] In the above Formulas 1a - 1d, * represents the position of attachment to the compound of Formula 1, n is 1, 2, or 3, R ３ and R ４ are, independently of one another, hydrogen; C １ -C ４ alkyl group, or they combine with the nitrogen atom to which they are attached to form piperidine or pyrrolidine (said piperidine or pyrrolidine being optionally substituted with C １ -C ４ alkyl). R ５ and R ６ are C １ to C ４ alkyl groups, or R ５ and R ６ form a ring to form cyclopentane, cyclohexane or tetrahydropyran, Cy is a nitrogen-containing heterocyclic group selected from the group consisting of homopiperazin-1-yl, piperidin-1-yl, piperidin-4-yl, morpholin-4-yl, and pyrrolidin-3-yl, and the nitrogen-containing heterocyclic group is C １ to C ４ optionally substituted with alkyl or benzyl. Claim 2 R １ The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is hydrogen; an isobutyl group; or a diethylaminoethyl group. Claim 3 R ２ The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R is a methyl group. Claim 4 The compound or a pharmaceutically acceptable salt thereof according to Claim 1, wherein A is a group of Formula 1a or a group of Formula 1b. Claim 5 R ３ and R ４ form a piperidine or pyrrolidine by bonding to the nitrogen atom to which they are attached, the compound according to claim 4 or a pharmaceutically acceptable salt thereof. Claim 6 The piperidine or pyrrolidine is C １ to C ４ substituted with alkyl, the compound according to claim 5 or a pharmaceutically acceptable salt thereof. Claim 7 N-(2-(Diethylamino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diisopropylamino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Dimethylamino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(3-(Diethylamino)propyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(4-Methylhomopiperazino)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(4-(1-Methyl)piperidinyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(4-Morpholino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(1-Pyrrolidino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(1-Piperidino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((1-(Dimethylamino)cyclopentyl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((1-(Dimethylamino)cyclohexyl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(3-(1-Benzyl)pyrrolidinyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(2-(1-Methyl)pyrrolidino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((S)-2-(1-Ethyl)pyrrolidinomethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(4-Morpholinoamino)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(1-Piperidinoamino)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diisopropylamino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diethylamino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Dimethylamino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(4-Morpholino)ethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((S)-2-(1-Ethyl)pyrrolidinomethyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diisopropylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diethylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Dimethylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(4-Morpholino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((S)-2-(1-Ethyl)pyrrolidinomethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; and N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride A compound selected from the group consisting of or a pharmaceutically acceptable salt thereof.

8. N-(2-(Diisopropylamino)ethyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-isobutoxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(Diisopropylamino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-(2-(4-Morpholino)ethyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride; N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(3-cyano-4-hydroxyphenyl)-4-methylthiazole-5-carboxamide hydrochloride; and N-((4-(Dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-2-(4-(2-diethylamino)ethoxy-3-cyanophenyl)-4-methylthiazole-5-carboxamide hydrochloride A compound selected from the group consisting of or a pharmaceutically acceptable salt thereof.

9. The compound of Chemical Formula 2 is reacted with an N-containing compound selected from the group consisting of NH ２ -(CH ２ ) ｎ -NR ３ R ４ , NH ２ -CH ２ -CR ５ R ６ -NR ３ R ４ , Cy, and NH ２ -Cy (wherein n, R ３ , R ４ , R ５ , and R ６ are the same as those defined in Claim 1; Cy is homopiperazine, and homopiperazine is optionally substituted with C １ -C ４ alkyl; NH ２ -Cy is a nitrogen-containing heterocyclic amine selected from the group consisting of piperidin-1-ylamine, piperidin-4-ylamine, morpholin-4-ylamine, and pyrrolidin-3-ylamine, and the nitrogen-containing heterocyclic amine is optionally substituted with C １ -C ４ alkyl or benzyl) to obtain a method for producing a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof, comprising an acylation step: [Chemical Formula 6] [Chemical Formula 7] wherein R １ , R ２ , and A are the same as those defined in claim 1.

10. The compound of Chemical Formula 3 is acylated using an N-containing compound selected from the group consisting of NH ２ -(CH ２ ) ｎ -NR ３ R ４ , NH ２ -CH ２ -CR ５ R ６ -NR ３ R ４ , Cy, and NH ２ -Cy (wherein n, R ３ , R ４ , R ５ , and R ６ are the same as those defined in Claim 1; Cy is homopiperazine, and homopiperazine is optionally substituted with C １ -C ４ alkyl; NH ２ -Cy is a nitrogen-containing heterocyclic amine selected from the group consisting of piperidin-1-ylamine, piperidin-4-ylamine, morpholin-4-ylamine, and pyrrolidin-3-ylamine, and the nitrogen-containing heterocyclic amine is optionally substituted with C １ -C ４ alkyl or benzyl) to prepare the compound of Chemical Formula 4; and A process for preparing a compound of Formula 1 or a pharmaceutically acceptable salt thereof, comprising alkylating a compound of Formula 4 with a compound of Formula 5: <Formula 5> 【Chemical 8】 【Chemical Formula 9】 【Chemical 10】

11. R １ X In the formula, R １ is a C1-C4 alkyl group; or a C1-C4 alkyl group substituted with mono- or di-C1-C5 alkylamino, R ２ and A are the same as those defined in claim 1, and X is a halogen. ​ A pharmaceutical composition for inducing autophagy, comprising, as an active ingredient, a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof in a therapeutically effective amount.

12. The pharmaceutical composition according to claim 11 for preventing or treating a neurodegenerative disease selected from the group consisting of Huntington's disease, Parkinson's disease, Alzheimer's disease, prion disease, multiple sclerosis, and Lou Gehrig's disease; a liver disease selected from the group consisting of liver fibrosis, cirrhosis, hepatitis, and fatty liver disease; a metabolic disease selected from the group consisting of diabetes, hyperlipidemia, obesity, and inflammation; or sepsis.

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