RU2795227C2 - New catechol derivatives or their salt, methods of their production and pharmaceutical compositions containing them - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein Y is O or S, (1) if Y is O, R₁ is hydrogen or a C₁-C₄ alkyl group substituted with mono- or di-C₁-C₅ alkylamino, R₂-C₁-C₆ alkyl group, R₃ is hydrogen, and R₄ - (4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl group, C₁-C₄ alkyl group substituted with mono- or di-C₁-C₅ alkylamino, C₁-C₄ alkyl group substituted with a nitrogen-containing cyclic ring selected from the group consisting of morpholine, piperidine or pyrrolidine, the nitrogen-containing cyclic ring being optionally substituted with C₁-C₄ alkyl, or a piperidinyl group optionally substituted with C₁-C₄ alkyl, or R₃ and R₄ are connected to each other by the nitrogen atom to which they are attached to form a piperazine ring, the piperazine ring being optionally substituted with C₁-C₄ alkyl, (2) if Y is S, R₁ and R₂ are independently hydrogen; C₁-C₆ alkyl group or C₁-C₄ alkyl group substituted with mono- or di-C₁-C₅ alkylamino, R₃ is hydrogen, and R₄-C₁-C₄ alkyl group substituted with mono- or di-C₁-C₅ alkylamino, or R₃ and R₄ are connected to each other by the nitrogen atom to which they are attached to form a piperazine ring, the piperazine ring being optionally substituted with C_1-C₄ alkyl. The invention also relates to methods for preparing a compound of formula 1 and its use.

EFFECT: new compounds have been obtained that can be used in medicine for the prevention, alleviation or treatment of neurodegenerative diseases, liver diseases, metabolic diseases by inducing autography.

16 cl, 4 dwg, 7 tbl, 26 ex

Formula (1)

Description

FIELD OF TECHNOLOGY

[1] This invention relates to novel derivatives of catechol or a pharmaceutically acceptable salt thereof, a process for their preparation, and a pharmaceutical composition containing them. More specifically, this invention relates to a novel derivative of catechol or a pharmaceutically acceptable salt thereof, having an alkyl moiety substituted with an alkylamino and/or an N-alkyl-substituted thiophene-(thio)carboxamide moiety, a process for their preparation, and a pharmaceutical composition containing them. The catechol derivative or pharmaceutically acceptable salt of the present invention has an excellent autophagy-inducing activity.

BACKGROUND OF THE INVENTION

[2] Autophagy, also referred to as autophagocytosis, is a naturally regulated cell mechanism that dismantles unnecessary or dysfunctional components. It provides an orderly breakdown and processing of cellular components. During autophagy, consumable components of the cytoplasm are isolated from the rest of the cell in a double-membrane vesicle known as the autophagosome. Next, the autophagosome fuses with the available lysosome, and as a result, the contents of the vesicle disintegrate and are processed. Generally, three forms of autophagy are described: macroautophagy, microautophagy, and chaperone-mediated autophagy (CMA). In disease, autophagy is seen as an adaptive response to stress that promotes cell survival; but in other cases it appears to promote cell death and complications. In acute cellular starvation, the breakdown of cellular components promotes cell survival by maintaining the energy level of the cell.

[3] At the same time, with a decrease in autophagy, various diseases can occur due to the accumulation of misfolded proteins, and so on. For example, it has been reported that autophagy induction can treat neurodegenerative diseases such as Huntington's disease (HD), Parkinson's disease (PD), Alzheimer's disease (AD), prion disease, multiple sclerosis, and amyotrophic lateral sclerosis (Lou Gehrig's disease) (e.g. patent Korea No. 10-1731908). It has also been reported that the induction of autophagy can treat liver diseases such as liver fibrosis, cirrhosis, hepatitis, and fatty liver disease (for example, Korea Publication No. 10-2017-0022790 published to the public). In addition, it has been reported that autophagy induction can treat metabolic diseases such as diabetes, hyperlipidemia, obesity, and inflammation (for example, Korea Publication No. 10-2018-0007307 published to the public). In addition, it has been reported that autophagy induction can inhibit excessive immune responses associated with sepsis (eg, Korea Publication No. 10-2012-0131401, published to the public).

[4] Therefore, it is contemplated that the autophagy-inducing compound can be successfully used to prevent, alleviate, or treat various autophagy-related diseases such as neurodegenerative diseases, liver diseases, metabolic diseases, sepsis, and so on.

DETAILED DISCLOSURE OF THE INVENTION

TECHNICAL PROBLEM

[5] The present inventors have found that a certain catechol derivative or a pharmaceutically acceptable salt thereof having an alkylamino substituted alkyl moiety and/or an N-alkyl-substituted thiophene-(thio)carboxamide moiety has excellent autophagy-inducing activity and, therefore, can be effectively applied to prevent, alleviate, or treat various autophagy-related diseases.

[6] The present invention provides said catechol derivative or pharmaceutically acceptable salt thereof, a method for their preparation, and a pharmaceutical composition containing them.

TECHNICAL SOLUTION

[7] One aspect of the present invention is a novel catechol derivative or pharmaceutically acceptable salt thereof.

[8] Another object of this invention is a method for obtaining the specified derivative compounds of catechol or its pharmaceutically acceptable salt.

[9] Another aspect of the present invention is a pharmaceutical composition containing said catechol derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

[10] Another aspect of the present invention is a method of treating an autophagy-related disease in a mammal in need of the treatment, comprising administering to the mammal an effective amount of said catechol derivative or a pharmaceutically acceptable salt thereof.

[11] Yet another object of the present invention is the use of said catechol compound derivative or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention, alleviation or treatment of an autophagy related disease.

TECHNICAL RESULTS OF THE INVENTION

[12] The compound of the present invention, i.e., a catechol derivative (hereinafter referred to as a derivative or compound) or a pharmaceutically acceptable salt thereof, having an alkyl moiety substituted with an alkylamino and/or an N-alkyl-substituted thiophene-(thio ) carboxamide moiety, has excellent autophagy inducing activity. Therefore, the compound or a pharmaceutically acceptable salt thereof according to the present invention can be successfully used to prevent, alleviate or treat various diseases associated with autophagy, including neurodegenerative diseases, liver diseases, metabolic diseases, sepsis, and so on. In particular, the catechol derivative or pharmaceutically acceptable salt thereof according to the present invention has a molecular structure capable of penetrating the blood-brain barrier, i.e. an alkyl-substituted amine moiety, such a structure can be used for the prevention, alleviation or treatment of diseases associated with cerebral blood flow, for example , neurodegenerative diseases such as Huntington's disease (HD), Parkinson's disease (PD), Alzheimer's disease (AD), prion disease, multiple sclerosis, Lou Gehrig's disease and the like.

DESCRIPTION OF FIGURES

[13] FIG. 1 shows the results of applying the experimental methods of Test Example 2 to evaluate liver function improvement activity by oral administration in a liver injury model.

[14] FIG. 2 shows the results of applying the experimental methods of Test Example 3 to evaluate liver function improvement activity by oral administration in a liver injury model.

[15] FIG. 3 shows the results obtained by staining with hematoxylin and eosin (H&E) of tissue samples obtained in Test Example 3. Scale mark: left black line 100 µm, right black line 100 µm.

[16] FIG. 4 shows the results obtained by Masson's trichrome staining of the tissue samples obtained in Test Example 3. Scale mark: left black line 100 µm, right black line 100 µm.

BEST EXAMPLE OF THE INVENTION

[17] As used herein, the term "alkyl" refers to a linear or branched aliphatic hydrocarbon radical. For example, C ₁ -C ₆ alkyl means a linear or branched aliphatic hydrocarbon containing from 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.

[18] The term "hydroxy" refers to the -OH radical. The term "alkoxy" refers to a radical formed by replacing a hydrogen atom of a hydroxy group with an alkyl. For example, C ₁ -C ₆ alkoxy contains methoxy, ethoxy, propoxy, n-butoxy, n-pentyloxy, isopropoxy, sec-butoxy, tert-butoxy, neopentyloxy, and isopentyloxy.

[19] The term "halogen" refers to a fluorine, bromine, chlorine or iodine radical.

[20] The term "amino" refers to the radical-NH ₂ . The term "alkylamino" refers to an amino formed by replacing the hydrogen atom(s) of an amino group with a mono- or dialkyl. For example, C ₁ -C ₆ alkylamino contains amino substituted with mono- or di-C _1-6 alkyl.

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

[22] Formula 1

[23]

[24] and

[25] Y is O or S,

[26] (1) if Y - O,

[27] R ₁ is hydrogen or a C ₁ -C ₄ alkyl group substituted with mono- or di-C ₁ -C ₅ alkylamino,

[28] R ₂ - C ₁ -C ₆ alkyl group,

[29] R ₃ is hydrogen, and R ₄ is (4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl group; C ₁ -C ₆ alkyl group; C ₁ -C ₄ alkyl group substituted with mono- or di-C ₁ -C ₅ alkylamino; a C ₁ -C ₄ alkyl group substituted with a nitrogen-containing cyclic ring (wherein the nitrogen-containing cyclic ring is optionally substituted with C ₁ -C ₄ alkyl); or a piperidinyl group optionally substituted with C ₁ -C ₄ alkyl, or

[30] R ₃ and R ₄ are connected to each other by the nitrogen atom to which they are attached to form a piperazine ring (wherein the piperazine ring is optionally substituted with C ₁ -C ₄ alkyl),

[31] (2) if Y is S,

[32] R ₁ and R ₂ are independently hydrogen; C ₁ -C ₆ alkyl group or C ₁ -C ₄ alkyl group substituted with mono- or di-C ₁ -C ₅ alkylamino,

[33] R ₃ is hydrogen and R ₄ is a C ₁ -C ₆ alkyl group or a C ₁ -C ₄ alkyl group substituted with mono- or di-C ₁ -C ₅ alkylamino, or

[34] R ₃ and R ₄ are connected to each other by the nitrogen atom to which they are attached to form a piperazine ring, the piperazine ring being optionally substituted with C ₁ -C ₄ alkyl.

[35] In a compound or pharmaceutically acceptable salt thereof according to this invention, Y may be O. Preferably, Y is O; R ₁ may be hydrogen or a diethylaminoethyl group, and R ₂ may be a methyl group. Also, the nitrogen-containing cyclic ring may be morpholine, piperidine or pyrrolidine.

[36] In one embodiment, the invention provides a compound, or a pharmaceutically acceptable salt thereof, wherein:

[37] Y is O,

[38] R ₁ is hydrogen or a diethylaminoethyl group,

[39] R ₂ - methyl group,

[40] R ₃ is hydrogen, and

[41] R ₄ - (4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl group; an isopropyl group, a dimethylaminoethyl group, a diethylaminoethyl group, a diisopropylaminoethyl group, a morpholinoethyl group optionally substituted with C ₁ -C ₄ alkyl, a piperidinoethyl group optionally substituted with C ₁ -C ₄ alkyl, a pyrrolidinoethyl group optionally substituted with C ₁ -C ₄ alkyl, or piperidinyl a group optionally substituted with C ₁ -C ₄ alkyl.

[42] In another embodiment, the invention provides a compound or a pharmaceutically acceptable salt thereof, wherein:

[43] Y is O,

[44] R ₁ is hydrogen or a diethylaminoethyl group,

[45] R ₂ - methyl group,

[46] R ₃ is hydrogen, and

[47] R ₄ - (4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl group.

[48] In yet another embodiment of the invention, a compound or a pharmaceutically acceptable salt thereof is provided, wherein:

[49] Y - O,

[50] R ₁ is hydrogen or a diethylaminoethyl group,

[51] R ₂ is a methyl group, and

[52] R ₃ and R ₄ are connected to each other by a nitrogen atom to which they are attached to form a piperazine ring, optionally substituted with C ₁ -C ₄ alkyl.

[53] In a compound or a pharmaceutically acceptable salt thereof according to this invention, Y may be S. Preferably, when Y is S, R ₁ may be hydrogen or a diethylaminoethyl group and R ₂ a methyl group.

[54] In one embodiment, the invention provides a compound, or a pharmaceutically acceptable salt thereof, wherein:

[55] Y is S,

[56] R ₁ is hydrogen or a diethylaminoethyl group,

[57] R ₂ - methyl group,

[58] R ₃ is hydrogen, and

[59] R ₄ is an isopropyl group or a diisopropylaminoethyl group.

[60] In another embodiment, the invention provides a compound, or a pharmaceutically acceptable salt thereof, wherein:

[61] Y is S,

[62] R ₁ is hydrogen or a diethylaminoethyl group,

[63] R ₂ is a methyl group, and

[64] R ₃ and R ₄ are connected to each other by a nitrogen atom to which they are attached to form a piperazine ring, optionally substituted with C ₁ -C ₄ alkyl.

[65] Preferably, the compound or a pharmaceutically acceptable salt thereof according to the present invention may be one or more compounds selected from the group consisting of:

[66] N-(4-methylpiperazino)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[67] N-(4-methylpiperazino)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[68] N-(4-methylpiperazino)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride,

[69] N-(4-methylpiperazino)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride,

[70] N-(2-(diisopropylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[71] N-(2-(diisopropylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[72] N-(2-(diisopropylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride,

[73] N-(2-(diisopropylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride,

[74] N-isopropyl-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[75] N-isopropyl-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride,

[76] N-isopropyl-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-thiocarboxamide,

[77] N-(2-(diethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[78] N-(2-(diethylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[79] N-(2-(dimethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[80] N-(2-(dimethylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxy-phenyl)thiophene-2-carboxamide hydrochloride,

[81] N-(2-(4-morpholino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[82] N-(2-(1-piperidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[83] N-(2-(1-piperidino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxy-phenyl)thiophene-2-carboxamide hydrochloride,

[84] N-(2-(1-pyrrolidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[85] N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[86] N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[87] N-(4-(1-methyl)piperidinyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[88] N-(2-(2-(1-methyl)pyrrolidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[89] N-(2-(2-(1-methyl)pyrrolidino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride,

[90] N-((S)-2-(1-ethyl)pyrrolidinomethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride and

[91] N-((S)-2-(1-ethyl)pyrrolidinomethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride.

[92] More preferably, the compound of the present invention is N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2- carboxamide or a pharmaceutically acceptable salt thereof (eg hydrochloride); or was N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide or a pharmaceutically acceptable thereof salt (eg hydrochloride).

[93] The compound of formula 1 according to this invention can be presented in the form of a pharmaceutically acceptable salt, for example, in the form of an acid addition salt. In particular, the compound of the present invention has an alkyl-substituted amine moiety, and therefore can be easily isolated in the form of an acid addition salt (for example, in the form of a hydrochloride) in contrast to conventional compounds (for example, the compounds described in Korean Publication No. 10-2017 -0022790). That is, the compound of the present invention in the form of an acid addition salt can be easily obtained by acid-base treatment processes, and can be easily applied to uniform expansion methods without performing column chromatography processes, unlike conventional compounds (for example, the compounds described in Korean Publication No. 10-2017-0022790). Also, the compound according to the present invention in the form of an acid addition salt has excellent solubility in water, and therefore can be easily prepared, and also provides excellent oral bioavailability. The acid addition salt may be derived from an inorganic acid or an organic acid such as hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid, phosphoric acid, acetic acid, lactic acid, citric acid, tartaric acid, succinic acid, maleic acid. acid, malonic acid, oxalic acid, fumaric acid, gluconic acid, sugar acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, pamoic acid, etc., without limitation to the above. The acid addition salt may be prepared by reacting a compound of Formula 1 with an inorganic acid or organic acid in a common solvent such as water, alcohol, tetrahydrofuran, acetone, or mixtures thereof.

[94] The compound of Formula 1, or a pharmaceutically acceptable salt thereof, may have substituent(s) containing an asymmetric carbon, and therefore may be in the form of a racemic mixture (RS) or in optical isomer forms such as (R) or (S) isomer. Therefore, unless otherwise indicated, a compound of Formula 1, or a pharmaceutically acceptable salt thereof, contains both the racemic mixture (RS) and optical isomers such as the (R) or (S) isomer. Also, the compound of Formula 1, or a pharmaceutically acceptable salt thereof, may be in the form of a cis or trans geometric isomer, depending on the substituent(s). Therefore, unless otherwise indicated, a compound of Formula 1, or a pharmaceutically acceptable salt thereof, contains both cis and trans geometric isomers. Also, the compound of Formula 1, or a pharmaceutically acceptable salt thereof, may be in the form of one or more diastereomeric isomers, or mixtures thereof. Therefore, unless otherwise indicated, a compound of Formula 1, or a pharmaceutically acceptable salt thereof, contains both the diastereomeric isomer(s) and a mixture thereof.

[95] The compound of Formula 1 or its pharmaceutically acceptable salt according to this invention may be in anhydrous form, in the form of a hydrate or in the form of a solvate. In addition, the compound of Formula 1 or its pharmaceutically acceptable salt according to this invention may be in amorphous or crystalline forms. Said amorphous or crystalline forms may also be in the form of a hydrate or in the form of a solvate. The hydrate or solvate may contain water or an organic solvent in a stoichiometric or non-stoichiometric amount relative to the compound of Formula 1 or a pharmaceutically acceptable salt thereof.

[96] This invention also includes a process for preparing a compound of Formula 1 or a pharmaceutically acceptable salt thereof.

[97] For example, a compound of Formula 1 or a pharmaceutically acceptable salt thereof, where Y is O (i.e., a compound of Formula 1a or a pharmaceutically acceptable salt thereof) can be prepared by acylation of a compound of Formula 4 with a compound of Formula 5 to obtain a compound of Formula 2; and coupling a compound of Formula 2 with a compound of Formula 3 to obtain a compound of Formula 1a as shown in the following Reaction Scheme 1:

[98] Reaction Scheme 1

[99]

[100] In Reaction Scheme 1, R ₁ , R ₂ , R ₃ and R ₄ are the same as defined above, Z is halogen and R ₆ and R ₇ are hydrogen; or connected to each other by the boron atom to which they are attached to form 4,4,5,5-tetramethyl-1,3,2-dioxaborolane.

[101] Compounds of Formulas 4 and 5, which are known compounds, are commercially available. The acylation can 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). The acylation can also be carried out by reacting a compound of Formula 4 with thionyl chloride, oxalyl chloride, phosphorus chloride, etc. to obtain an acyl chloride followed by reaction with a compound of Formula 5. The acylation can be carried out in a common organic solvent such as dichloromethane.

[102] Coupling can be performed in the presence of a catalyst (eg tetrakis(triphenylphosphine)palladium(0)) and a base (eg sodium carbonate). The reaction of a compound of Formula 2 with a compound of Formula 3 can be carried out at a molar ratio ranging from 1:2 to 2:1, preferably at a molar ratio of about 1:1. Coupling can be carried out in water, C ₁ -C ₄ alcohol, tetrahydrofuran, 1,2-dimethoxyethane, or mixtures thereof.

[103] In addition, a compound of Formula 1 or a pharmaceutically acceptable salt thereof, wherein Y is S (i.e., a compound of Formula 1b or a pharmaceutically acceptable salt thereof) can be prepared by performing thioamidation of a compound of Formula 2 to obtain a compound of Formula 6 and coupling a compound of Formula 6 6 with a Formula 3 compound to give a Formula 1b compound as shown in the following Reaction Scheme 2:

[104] Reaction Scheme 2

[105]

[106] In Reaction Scheme 2, R ₁ , R ₂ , R ₃ , R ₄ , Z, R ₆ and R ₇ are the same as defined above.

[107] In Reaction Scheme 2, a compound of Formula 2 can be prepared as described for Reaction Scheme 1. Thioamidation can be carried out via reaction of a compound of Formula 2 with P ₄ S ₁₀ , bis(tricyclohexyltin) sulfide, or Lawesson's reagent. The thioamidation reaction can be carried out in toluene, dichloromethane, tetrahydrofuran, or a mixed solvent thereof. In addition, the coupling reaction can be carried out as described in Reaction Scheme 1 using a compound of Formula 6 instead of a compound of Formula 2.

[108] And also, a compound of Formula 1 or a pharmaceutically acceptable salt thereof, where Y is S (i.e., a compound of Formula 1b or a pharmaceutically acceptable salt thereof) can be obtained by thioamidation of a compound of Formula 1a to obtain a compound of Formula 1b, as shown in the following reaction scheme 3:

[109] Reaction Scheme 3

[110]

[111] In Reaction Scheme 3, R ₁ , R ₂ , R ₃ , and R ₄ are the same as defined above.

[112] In Reaction Scheme 3, a compound of Formula 1a can be prepared as described in Reaction Scheme 1. In addition, thioamidation can be performed as described in Reaction Scheme 2 using a compound of Formula 1a instead of a compound of Formula 2.

[113] The catechol derivative of the present invention, that is, the compound of Formula 1 or a pharmaceutically acceptable salt thereof, has excellent autophagy-inducing activity. Therefore, the compound of Formula 1 or a pharmaceutically acceptable salt thereof according to the present invention can be successfully used to prevent, alleviate or treat various autophagy-related diseases, including neurodegenerative diseases, liver diseases, metabolic diseases, sepsis, and so on.

[114] Therefore, the present invention comprises a pharmaceutical composition for inducing autophagy containing a therapeutically effective amount of a compound of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[115] Diseases associated with autophagy include, without limitation, various diseases that can be prevented, alleviated or cured by induction of autophagy. For example, the pharmaceutical composition of the present invention may be a pharmaceutical composition for preventing, alleviating, or treating neurodegenerative diseases selected from the group consisting of Huntington's disease, Parkinson's disease, Alzheimer's disease, prion disease, multiple sclerosis, and Lou Gehrig's disease; liver diseases selected from the group consisting of liver fibrosis, liver cirrhosis, hepatitis, and fatty liver disease; metabolic diseases selected from the group consisting of diabetes, hyperlipidemia, obesity and inflammation, or sepsis.

[116] The pharmaceutical composition according to this invention may contain a pharmaceutically acceptable carrier such as diluents, disintegrants, sweeteners, lubricants or flavoring agents. The pharmaceutical composition may be in an oral form such as tablets, capsules, powders, granules, suspensions, emulsions or syrups; or in a parenteral dosage form such as external solutions, external suspensions, external emulsions, gels (eg ointments), inhalers, sprays, injections, according to conventional methods. The dosage form may be in various forms, such as dosage forms for single use or for multiple use.

[117] The pharmaceutical composition of the present invention may contain, for example, a diluent (eg, lactose, corn starch, etc.), a lubricant (eg, magnesium stearate), an emulsifying agent, a suspending agent, a stabilizer, and/or an isotonic agent. If necessary, the composition additionally contains sweeteners and/or flavoring agents.

[118] the Composition according to this invention can be used orally or parenterally, including by inhalation, intravenous, intraperitoneal, subcutaneous, intracerebroventricular, rectal and topical application. Thus, the composition according to this invention can be included in various forms such as tablets, capsules, aqueous solutions or suspensions. In the case of oral tablets, carriers such as lactose, corn starch and lubricants such as magnesium stearate are commonly used. In the case of oral capsules, lactose and/or dried cornstarch can be used as a diluent. If an aqueous suspension is required for oral administration, the active ingredient may be combined with emulsifying and/or suspending agents. Certain sweeteners and/or flavoring agents may be used, if desired. For intramuscular, intraperitoneal, subcutaneous and intravenous administration, sterile solutions of the active ingredient are usually prepared; however, it is necessary to adjust the pH of these solutions accordingly and buffer them. To make the drug isotonic, it is necessary to control the total concentration of dissolved substances for intravenous administration. The composition according to this invention may be in the form of an aqueous solution containing pharmaceutically acceptable carriers, such as saline, the pH of which is 7.4. These solutions can be injected into the patient's bloodstream intramuscularly by local bolus injection.

[119] The catechol derivative of this invention, that is, the compound of Formula 1 or a pharmaceutically acceptable salt thereof, may be administered to a patient in a therapeutically effective amount in the range of about 0.0001 mg/kg to about 100 mg/kg per day, preferably from about 0.001 mg/kg up to about 100 mg/kg per day. The introduction can be performed orally or parenterally one or more times a day. Of course, the dose may be changed depending on the patient's age, condition, body weight, susceptibility, degree of disease, route of administration, duration of administration, and the like. Depending on the route of administration, the pharmaceutical composition according to the invention may contain a compound of 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.

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

[121] This invention also includes the use of a compound of Formula 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for inducing autophagy in a mammal in need thereof. For example, this invention includes the use of a compound of Formula 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention, amelioration or treatment of neurodegenerative diseases selected from the group consisting of Huntington's disease, Parkinson's disease, Alzheimer's disease, prion disease, multiple sclerosis and Lou Gehrig's disease. ; liver diseases selected from the group consisting of liver fibrosis, liver cirrhosis, hepatitis, and fatty liver disease; metabolic diseases selected from the group consisting of diabetes, hyperlipidemia, obesity and inflammation; or sepsis.

[122] The following examples and test cases are for illustrative purposes only and are not intended to limit the scope of the invention.

[123] The analysis of the compounds obtained in the examples below was performed as described below: analysis of the spectrum of nuclear magnetic resonance (NMR) was performed on a Bruker 400 MHz spectrometer and the results of the analysis of chemical shifts are expressed in µg/g (ppm). Column chromatography was performed on silica gel (Merck, 70-230 mesh). Unless otherwise noted, all starting materials were purchased commercially and used without further purification. All reactions and chromatographic fractions were analyzed by thin layer chromatography (TLC) on a 250 nm silica gel plate and visualized by UV or iodine (I ₂ ) staining.

[124]

[125] Example 1: Preparation of N-(4-methylpiperazino)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[126]

[127] A mixture of 5-bromothiophene-2-carboxylic acid (6.21 g) with dichloromethane (30 ml) and dimethylformamide (0.25 ml) was stirred for 10 minutes. Thionyl chloride (2.64 ml) was added to the mixture, after which the mixture was heated at reflux with stirring for 3 hours. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. Dichloromethane (45 ml) was added to the resulting concentrate, which was then cooled to 0 ~ 10 °C. K ₂ CO ₃ (4.62 g) was added to the reaction mixture, after which the mixture was stirred for 20 minutes. N-methylpiperazine (3.30 ml) was added to the mixture. The reaction mixture was stirred at room temperature for 2 hours, after which purified water (45 ml) was added thereto. The resulting wash water was extracted with dichloromethane (30 ml). The extract was combined with the reaction mixture. The resulting mixture was washed with purified water (30 ml), dried over anhydrous sodium sulfate, concentrated in vacuo to give 8.66 g of intermediate, i.e. N-(4-methylpiperazino)-5-bromo-thiophene-2-carboxamide (yield: 100 %).

[128] To N-(4-methylpiperazino)-5-bromo-thiophene-2-carboxamide (8.66 g) was added tetrakis(triphenylphosphine)palladium(0) (3.48 g) and 1,2-dimethoxyethane (51 ml). To the mixture was added a solution of sodium carbonate (9.36 g) in purified water (51 ml), after which the mixture was stirred at room temperature for 30 minutes. A solution of (4-hydroxy-3-methoxyphenyl)-(tetramethyl-1,3-oxa)borolane (8.25 g) in ethanol (51 ml) was added to the reaction mixture, after which the mixture was stirred at a temperature of about 80 °C for 5 o'clock. The reaction mixture was cooled to room temperature, after which it was filtered to remove insoluble material. The resulting insoluble material was washed with ethanol (40 ml), after which the resulting washing solution was combined with the filtrate. The resulting filtrate was concentrated under reduced pressure to remove the solvent. Purified water (200 ml) and 6N hydrochloric acid (10 ml) were added to the resulting residue with stirring. The resulting solution was washed twice with chloroform (100 ml and 50 ml, respectively), after which its pH was adjusted to pH 8 ~ 9 using sodium hydroxide (about 3.0 g). The solution was extracted twice with chloroform (100 ml and 50 ml, respectively). The combined extract was dried over anhydrous sodium sulfate and then concentrated in vacuo. Acetone (30 ml) was added to the resulting concentrate. The mixture was stirred for 1 hour, after which it was filtered. The resulting solid was washed with acetone (5 ml), dried in vacuo at 30°C for 3 hours to give N-(4-methylpiperazino)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide (8, 68 g, yield: 87.0%). The dried solid (1.50 g) was dissolved in a mixed solvent of methanol (10 ml) and chloroform (3 ml), after which a 2N hydrochloric acid solution in ethanol (1.5 ml) was added thereto. The mixture was concentrated under reduced pressure. Acetone (10 ml) was added to the resulting concentrate. The mixture was stirred for 1 hour, after which it was filtered. The resulting solid was washed with acetone (5 ml) and then dried in vacuo to obtain 1.52 g of the title compound (yield: 91.6%, overall yield: 80.0%).

[129] TLC R _f = 0.20 in 10% MeOH in chloroform.

[130] ¹ H NMR (400MHz, MeOH-d ₄ ) δ 7.46 (d, 1H, J = 4.0 Hz), 7.31 (d, 1H, J = 4.0 Hz), 7.22 (d, 1H, J = 2.0 Hz), 7.16 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 6.98 (d, 1H, J = 8.0 Hz), 4.72~4.60(m, 2H), 3.93(s, 3H), 3.70~3.48(m, 4H), 3.30~3.20(m, 2H), 2, 99(s, 3H)

[131]

[132] Example 2: Preparation of N-(4-methylpiperazino)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[133]

[134] To a mixture of N-(4-methylpiperazino)-5-(4-hydroxy-3-methoxy-phenyl)thiophene-2-carboxamide (3.32 g) obtained by the same procedures as in Example 1, sodium hydroxide (0.80 g) was added with 2-diethylaminoethyl chloride hydrochloride (1.72 g) and toluene (50 ml). The reaction mixture was stirred at 85°C for 4 hours, after which it was cooled to room temperature. Purified water (50 ml) was added to the reaction mixture, after which the mixture was stirred for 30 minutes. The separated organic layer was washed with saturated sodium carbonate solution and then extracted with 1N hydrochloric acid (50 ml). The extract was washed with ethyl acetate (20 ml), after which its pH was adjusted to pH 7~8 using sodium hydroxide (about 2.0 g). The solution was extracted twice with dichloromethane (50 ml and 30 ml, respectively). The resulting extract was dried over anhydrous sodium sulfate and then concentrated in vacuo to give 4.0 g of crude product, i.e. N-(4-methylpiperazino)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thio- phen-2-carboxamide (yield: 89.5%). To the resulting residue was added a 4N solution of hydrochloric acid in ethanol (6.0 ml) and acetone (60 ml). The mixture was stirred at room temperature for 1 hour, after which it was filtered. The resulting solid was washed with acetone (5 ml) and then dried in vacuo to obtain 4.08 g of the title compound (yield: 87.4%, overall yield: 78.2%).

[135] TLC R _f = 0.21 in 10% MeOH in chloroform.

[136] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.50 (d, 1H, J = 4.0 Hz), 7.41 (d, 1H, J = 4.0 Hz), 7.33 (d, 1H, J = 2.0 Hz), 7.30 (dd, 1H, J = 2.0 Hz, 8.4 Hz), 7.13 (d, 1H, J = 8.4 Hz), 4.72 ~ 4.60 (m, 2H), 4.42 (t, 2H, J = 4.8Hz), 3.97 (s, 3H), 3.70 ~ 3.60 (m, 6H), 3, 50~3.38(m, 4H), 3.30~3.20(m, 2H), 2.99(s, 3H), 1.42(t, 6H, J=7.2Hz).

[137]

[138] Example 3: Preparation of N-(4-methylpiperazino)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride

[139]

[140] A mixture of N-(4-methylpiperazino)-5-bromo-thiophene-2-carboxamide (2.92 g), obtained by the same procedures as in Example 1, with toluene (15 ml) and tetrahydrofuran (15 ml) was stirred for 10 minutes. Lawesson's reagent (4.25 g) was added to the mixture, which was then stirred at 50°C for 5 hours. The reaction mixture was cooled to room temperature, after which it was filtered to remove insoluble material. Ethyl acetate (200 ml) was added to the resulting filtrate, followed by extraction with a mixed solution of purified water (200 ml) and 2N hydrochloric acid (25 ml) and a mixed solution of purified water (200 ml) and 2N hydrochloric acid (15 ml), respectively. The combined extract was washed with ethyl acetate (200 ml) and then adjusted to pH 9~10 using sodium hydroxide (ca. 4.3 g). The solution was extracted three times with chloroform (100 ml, 50 ml and 50 ml, respectively). The resulting extract was dried over anhydrous sodium sulfate, and then concentrated in vacuo to obtain 2.47 g of an intermediate, i.e., N-(4-methylpiperazino)-5-bromo-thiophene-2-thiocarboxamide (yield: 80.0%).

[141] To N-(4-methylpiperazino)-5-bromo-thiophene-2-carboxamide (2.47 g) was added tetrakis(triphenylphosphine)palladium(0) (1.04 g) and 1,2-dimethoxyethane (15 ml). To the mixture was added a solution of sodium carbonate (2.81 g) in purified water (15 ml), after which the mixture was stirred at room temperature for 30 minutes. A solution of (4-hydroxy-3-methoxyphenyl)-(tetramethyl-1,3-oxa)borolane (2.48 g) in ethanol (15 ml) was added to the reaction mixture, after which the mixture was stirred at a temperature of about 80°C for 5 o'clock. The reaction mixture was cooled to room temperature, after which it was filtered to remove insoluble material. The resulting filtrate was washed with ethanol (15 ml) and then concentrated under reduced pressure to remove the solvent. Purified water (60 ml) and 6N hydrochloric acid (3 ml) were added to the resulting residue with stirring. The resulting solution was washed twice with chloroform (30 ml and 15 ml, respectively), after which its pH was adjusted to pH 8 ~ 9 using sodium hydroxide (about 1.0 g). The solution was extracted twice with chloroform (30 ml and 50 ml, respectively). The resulting extract was dried over anhydrous sodium sulfate and then concentrated in vacuo. Acetone (10 ml) was added to the resulting concentrate. The mixture was stirred for 1 hour, after which it was filtered. The resulting solid (i.e., N-(4-methylpiperazino)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-thiocarboxamide) was dissolved in a mixed solvent of methanol (50 ml) and chloroform (15 ml), after which a 2N solution of hydrochloric acid in ethanol (5 ml) was added thereto. The mixture was concentrated under reduced pressure. Acetone (50 ml) was added to the resulting concentrate. The mixture was stirred for 1 hour, after which it was filtered. The resulting solid was washed with acetone (5 ml) and then dried in vacuo to obtain 2.49 g of the title compound (yield: 80.0%).

[142] TLC R _f = 0.33 in 10% MeOH in chloroform.

[143] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.30 (d, 1H, J = 3.6 Hz), 7.21~7.15 (m, 3H), 6.98 (d , 1H, J=8.0Hz), 5.20~5.10(m, 2H), 3.93(s, 3H), 3.85~3.65(m, 4H), 3.32~ 3.25(m, 2H), 3.00(s, 3H)

[144]

[145] Example 4: Preparation of N-(4-methylpiperazino)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride

[146]

[147] A mixture of N-(4-methylpiperazino)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide (4.0 g) obtained by the same procedures as in Example 2, with toluene (15 ml) and tetrahydrofuran (15 ml) was stirred for 10 minutes. Lawesson's reagent (4.04 g) was added to the reaction mixture, which was then stirred at 50°C for 5 hours. The reaction mixture was cooled to room temperature, after which it was filtered to remove insoluble material. Chloroform (100 ml) was added to the resulting filtrate, followed by extraction with a solution of purified water (60 ml) and 2N hydrochloric acid (40 ml). The resulting extract was washed with chloroform (30 ml), after which its pH was adjusted to pH 8~9 using sodium hydroxide (about 4.4 g). The solution was extracted three times with chloroform (100 ml, 50 ml and 50 ml, respectively). The resulting extract was dried over anhydrous sodium sulfate and then concentrated in vacuo. To the resulting residue was added a 4N solution of hydrochloric acid in ethanol (6.0 ml) and acetone (60 ml). The mixture was stirred at room temperature for 1 hour, after which it was filtered. The resulting solid was washed with acetone (5 ml) and then dried in vacuo to obtain 4.0 g of the title compound (yield: 83.0%).

[148] TLC R _f = 0.29 in 10% MeOH in chloroform.

[149] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.35 (d, 1H, J = 3.6 Hz), 7.32 ~ 7.26 (m, 3H), 7.12 (d , 1H, J=8.4Hz), 5.25~5.15(m, 2H), 4.41(t, 2H, J=4.8Hz), 3.97(s, 3H), 3 .85 ~ 3.75 (m, 2H), 3.66 (t, 4H, J = 4.8Hz), 3.50 ~ 3.35 (m, 6H), 3.01 (s, 3H), 1.42 (t, 3H, J = 7.2 Hz)

[150]

[151] Example 5: Preparation of N-(2-(diisopropylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[152]

[153] The title compound was obtained by the same procedures as in Example 1 using 2-(diisopropylamino)ethylamine instead of N-methylpiperazine (yield: 80.0%).

[154] TLC R _f = 0.23 in 10% MeOH in chloroform.

[155] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.70 (d, 1H, J = 4.0 Hz), 7.34 (d, 1H, J = 4.0 Hz), 7, 23 (d, 1H, J = 2.0 Hz), 7.18 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 6.86 (d, 1H, J = 8.4 Hz) , 3.94 (s, 3H), 3.85 (p, 2H, J = 6.4 Hz), 3.73 (t, 2H, J = 6.4 Hz), 3.38 (t, 2H, J = 6.4Hz), 1.47 ~ 1.42 (m, 12H)

[156]

[157] Example 6: Preparation of N-(2-(diisopropylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[158]

[159] To a mixture of N-(2-(diisopropylamino)ethyl)-5-bromo-thiophene-2-carboxamide (4.00 g) (an intermediate compound obtained by the same procedures as in Example 5), with tetrakis (triphenylphosphine)palladium(0) (1.39 g) and 1,2-dimethoxyethane (20 ml) was added a solution of sodium carbonate (3.80 g) in purified water (20 ml). The resulting mixture was stirred at room temperature for 30 minutes, after which a solution of (4-(2-diethylamino)ethoxy-3-methoxyphenyl)-(tetramethyl-1,3-oxa)borolane (4.19 g) in ethanol was added thereto. (20 ml). The reaction mixture was stirred at about 80°C for 5 hours, cooled to room temperature, and then filtered to remove insoluble material. The resulting filtrate was washed with ethanol (40 ml) and then concentrated under reduced pressure to remove the solvent. 2N hydrochloric acid (70 ml) was added to the resulting residue with stirring. The resulting solution was washed twice with ethyl acetate (50 ml), after which its pH was adjusted to pH 8 ~ 9 using sodium hydroxide (about 6.0 g). The solution was extracted twice with dichloromethane (100 ml and 50 ml, respectively). The combined extract was dried over anhydrous sodium sulfate and then concentrated in vacuo. A 2N solution of hydrochloric acid in ethanol (1.0 ml) was added to the resulting concentrate. The mixture was concentrated under reduced pressure. Acetone (60 ml) was added to the resulting concentrate. The mixture was stirred for 1 hour, after which it was filtered. The resulting solid was washed with acetone (5 ml) and then dried in vacuo to obtain 5.53 g of the title compound (yield: 83.0%).

[160] TLC R _f = 0.15 in 10% MeOH in chloroform.

[161] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.76 (d, 1H, J = 4.0 Hz), 7.43 (d, 1H, J = 4.0 Hz), 7.33 (d, 1H, J = 2.0 Hz), 7.31 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 7.13 (d, 1H, J = 8.0 Hz), 4.42 (t, 2H, J = 4.8 Hz), 3.97 (s, 3H), 3.85 (p, 2H, J = 6.4 Hz), 3.75 (t, 2H, J = 6, 4Hz), 3.66(t, 2H, J=4.8Hz), 3.48~3.36(m, 6H), 1.45(dd, 12H, J=2.4Hz, 6, 4 Hz), 1.42 (t, 6H, J = 7.2 Hz)

[162]

[163] Example 7: Preparation of N-(2-(diisopropylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride

[164]

[165] The title compound was prepared following the same procedures as in Example 4 using N-(2-(diisopropylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide prepared according to the same procedures as in Example 5 (yield: 75.0%).

[166] TLC R _f = 0.30 in 10% MeOH in chloroform.

[167] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.58 (d, 1H, J = 4.0 Hz), 7.24 (d, 1H, J = 4.0 Hz), 7, 18~7.08(m, 2H), 6.76(d, 1H, J=8.0Hz), 4.25(t, 2H, J=6.4Hz), 3.81(p, 2H , J=6.4Hz), 3.96(s, 3H), 3.38(t, 2H, J=6.4Hz), 1.45~1.38(m, 12H)

[168]

[169] Example 8: Preparation of N-(2-(diisopropylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride

[170]

[171] The title compound was prepared following the same procedures as in Example 4 using N-(2-(diisopropylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2 -carboxamide obtained by the same procedures as in Example 6 (yield: 80.0%).

[172] TLC R _f = 0.25 in 10% MeOH in chloroform.

[173] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.66 (d, 1H, J = 4.0 Hz), 7.40 (d, 1H, J = 4.0 Hz), 7, 30~7.25(m, 2H), 7.11(d, 1H, J=8.0Hz), 4.40(t, 2H, J=4.8Hz), 4.13(t, 2H , J = 6.4 Hz), 3.96 (s, 3H), 3.83 (p, 2H, J = 6.4 Hz), 3.63 (t, 2H, J = 4.8 Hz), 3.45~3.33(m, 6H), 1.42(dd, 12H, J=2.4Hz, 6.4Hz), 1.38(t, 6H, J=7.2Hz)

[174]

[175] Example 9: Preparation of N-isopropyl-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[176]

[177] The title compound was obtained by the same procedures as in Example 2, using isopropylamine instead of N-methylpiperazine (yield: 73.0%).

[178] TLC R _f = 0.20 in 10% MeOH in chloroform.

[179] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.68 (d, 1H, J = 4.0 Hz), 7.36 (d, 1H, J = 4.0 Hz), 7, 32 (d, 1H, J = 2.4 Hz), 7.29 (dd, 1H, J = 2.4 Hz, 8.4 Hz), 7.11 (d, 1H, J = 8.4 Hz) , 4.40 (t, 2H, J = 4.8 Hz), 4.20 (p, 1H, J = 6.8 Hz), 3.97 (s, 3H), 3.65 (t, 2H, J=4.8Hz), 3.46~3.37(m, 4H), 1.41(t, 6H, J=7.2Hz), 1.28(d, 6H, J=6.8 Hz)

[180]

[181] Example 10: Preparation of N-isopropyl-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride

[182]

[183] The title compound was prepared following the same procedures as in Example 4 using N-isopropyl-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide prepared according to the same procedures as in Example 9 (yield: 70.0%).

[184] TLC R _f = 0.26 in 10% MeOH in chloroform.

[185] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.56 (d, 1H, J = 4.0 Hz), 7.33 (d, 1H, J = 4.0 Hz), 7, 30~7.25(m, 2H), 7.08(d, 1H, J=8.0Hz), 4.80(p, 1H, J=6.8Hz), 4.38(t, 2H , J=4.8Hz), 3.95(s, 3H), 3.62(t, 2H, J=4.8Hz), 3.45~3.35(m, 4H), 1.39 (t, 6H, J = 7.2 Hz), 1.33 (d, 6H, J = 6.8 Hz)

[186]

[187] Example 11: Preparation of N-isopropyl-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-thiocarboxamide

[189] Intermediate (i.e. N-isopropyl-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide) (3.5 g) was obtained by the same procedures as in Example 1, using isopropylamine instead of N-methylpiperazine (Yield: 85.0%). Toluene (50 ml) was added thereto, followed by stirring for 10 minutes. Lawesson's reagent (5.0 g) was added to the mixture, which was then stirred at approximately 80°C for 5 hours. The reaction mixture was cooled to room temperature, after which it was filtered to remove insoluble material. The resulting filtrate was concentrated in vacuo. The resulting residue was purified by column chromatography (ethyl acetate/hexane=1/3, v/v), and then concentrated under reduced pressure. The resulting concentrate was stirred in a mixed solvent of ethyl acetate and hexane (1/3 (v/v), 10 ml) for 1 hour, after which it was filtered. The resulting solid was washed with hexane (5 ml) and then dried at 40°C for 5 hours to obtain 2.4 g of the title compound (yield: 65.0%).

[190] TLC R _f = 0.35 in 10% MeOH in chloroform.

[191] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.53 (d, 1H, J = 4.0 Hz), 7.24 (d, 1H, J = 4.0 Hz), 7, 19 (d, 1H, J = 2.0 Hz), 7.14 (dd, 1H, J = 2.0 Hz, 8.4 Hz), 6.82 (d, 1H, J = 8.4 Hz) , 4.80 (p, 1H, J = 6.8 Hz), 3.92 (s, 3H), 1.32 (d, 6H, J = 6.8 Hz)

[192]

[193] Example 12: Preparation of N-(2-(diethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[194]

[195] The title compound was obtained by the same procedures as in Example 1 using 2-(diethylamino)ethylamine instead of N-methylpiperazine (yield: 34.8%).

[196] TLC R _f = 0.13 in 10% MeOH in chloroform.

[197] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.71 (d, 1H, J = 4.0 Hz), 7.32 (d, 1H, J = 4.0 Hz), 7, 22 (d, 1H, J = 2.0 Hz), 7.17 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 6.86 (d, 1H, J = 8.4 Hz) , 3.94(s, 3H), 3.76(t, 2H, J=6.0Hz), 3.41(t, 2H, J=6.0Hz), 3.38~3.34( m, 4H), 1.38 (t, 6H, J = 7.2 Hz)

[198]

[199] Example 13: Preparation of N-(2-(diethylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[200]

[201] To a solution of sodium hydroxide (0.96 g) in a mixed solvent of purified water (10 ml) and tetrahydrofuran (50 ml) was added N-(2-(diethylamino)ethyl)-5-(4-hydroxy-3- methoxyphenyl)thiophene-2-carboxamide (2.21 g), obtained by the same procedures as in Example 12, and 2-diethylaminoethyl chloride hydrochloride (0.99 g). The reaction mixture was stirred at a temperature of about 70°C for 4 hours, cooled to room temperature, and then separated into an aqueous layer and an organic layer. The resulting organic layer was concentrated under reduced pressure, followed by extraction with 1N hydrochloric acid (20 ml). The resulting extract was washed with ethyl acetate (20 ml), after which its pH was adjusted to pH 8 ~ 9 using sodium hydroxide (about 0.9 g). The solution was extracted twice with dichloromethane (50 ml and 30 ml, respectively). The resulting extract was dried over anhydrous sodium sulfate and then concentrated in vacuo to give the crude product, N-(2-(diethylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide . Ethanol (15 ml) was added to the resulting residue. To the resulting solution was added a solution of 1N hydrochloric acid in ether (15 ml). The mixture was heated for 1 hour and then concentrated. Acetone (50 ml) was added to the resulting residue. The mixture was stirred at room temperature for 30 minutes, after which it was filtered. The resulting solid was washed with acetone (5 ml) and then dried in vacuo to obtain 2.20 g of the title compound (yield: 73.3%).

[202] TLC R _f = 0.23 in 20% MeOH in chloroform.

[203] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.75 (d, 1H, J = 4.0 Hz), 7.42 (d, 1H, J = 4.0 Hz), 7.33 ( d, 1H, J = 2.0 Hz), 7.31 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 7.12 (d, 1H, J = 8.0 Hz), 4 .42 (t, 2H, J = 4.8 Hz), 3.97 (s, 3H), 3.77 (t, 2H, J = 6.0 Hz), 3.66 (t, 2H, J = 6.0Hz), 3.50 ~ 3.35(m, 10H), 1.42(t, 6H, J=7.2Hz), 1.39(t, 6H, J=7.2Hz)

[204]

[205] Example 14: Preparation of N-(2-(dimethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[206]

[207] The title compound was obtained by the same procedures as in Example 1 using 2-(dimethylamino)ethylamine instead of N-methylpiperazine (yield: 64.5%).

[208] TLC R _f = 0.12 in 10% MeOH in chloroform.

[209] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.69 (d, 1H, J = 4.0 Hz), 7.32 (d, 1H, J = 4.0 Hz), 7, 23 (d, 1H, J = 2.0 Hz), 7.17 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 6.86 (d, 1H, J = 8.0 Hz) , 3.94 (s, 3H), 3.76 (t, 2H, J = 6.0 Hz), 3.40 (t, 2H, J = 6.0 Hz), 3.01 (s, 6H)

[210]

[211] Example 15: Preparation of N-(2-(dimethylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[212]

[213] The title compound was prepared following the same procedures as in Example 13 using N-(2-(dimethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide prepared according to by the same procedures as in Example 14 instead of N-(2-(diethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide (yield: 54.7%).

[214] TLC R _f = 0.34 in 20% MeOH in chloroform.

[215] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.77 (d, 1H, J = 4.0 Hz), 7.41 (d, 1H, J = 4.0 Hz), 7.33 ( d, 1H, J = 2.0 Hz), 7.30 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 7.12 (d, 1H, J = 8.4 Hz), 4 .42 (t, 2H, J = 4.8 Hz), 3.97 (s, 3H), 3.78 (t, 2H, J = 6.0 Hz), 3.66 (t, 2H, J = 6.0Hz), 3.50~3.35(m, 6H), 3.01(s, 6H), 1.42(t, 6H, J=7.2Hz)

[216]

[217] Example 16: Preparation of N-(2-(4-morpholino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[218]

[219] The title compound was obtained by the same procedures as in Example 1, using 4-(2-aminoethyl)morpholine instead of N-methylpiperazine (yield: 16.8%).

[220] TLC R _f = 0.32 in 10% MeOH in chloroform.

[221] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.71 (d, 1H, J = 4.0 Hz), 7.33 (d, 1H, J = 4.0 Hz), 7, 23 (d, 1H, J = 2.0 Hz), 7.17 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 6.86 (d, 1H, J = 8.0 Hz) , 4.14 ~ 4.09 (m, 2H), 3.94 (s, 3H), 3.87 ~ 3.77 (m, 4H), 3.74 ~ 3.69 (m, 2H), 3 .43 (t, 2H, J = 5.6Hz), 3.28 ~ 3.23 (m, 2H)

[222]

[223] Example 17: Preparation of N-(2-(1-piperidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[224]

[225] The title compound was obtained by the same procedures as in Example 1 using 1-(2-aminoethyl)piperidine instead of N-methylpiperazine (yield: 52.7%).

[226] TLC R _f = 0.15 in 10% MeOH in chloroform.

[227] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.69 (d, 1H, J = 4.0 Hz), 7.33 (d, 1H, J = 4.0 Hz), 7, 23 (d, 1H, J = 2.0 Hz), 7.17 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 6.86 (d, 1H, J = 8.4 Hz) , 3.94(s, 3H), 3.76(t, 2H, J=6.0Hz), 3.75~3.67(m, 2H), 3.38~3.35(m, 2H ), 3.02 (t, 2H, J = 12Hz), 2.03 ~ 1.95 (m, 2H), 1.92 ~ 1.77 (m, 3H), 1.63 ~ 1.52 ( m, 1H)

[228]

[229] Example 18: Preparation of N-(2-(1-piperidino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[230]

[231] The title compound was prepared following the same procedures as in Example 13 using N-(2-(1-piperidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide, obtained by the same procedures as in Example 17, instead of N-(2-(diethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide (yield: 82.6%).

[232] TLC R _f = 0.27 in 20% MeOH in chloroform.

[233] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.75 (d, 1H, J = 4.0 Hz), 7.42 (d, 1H, J = 4.0 Hz), 7, 33 (d, 1H, J = 2.0 Hz), 7.31 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 7.12 (d, 1H, J = 8.4 Hz) , 4.42(t, 2H, J=4.8Hz), 3.97(s, 3H), 3.78(t, 2H, J=6.0Hz), 3.75~3.67(m, 2H), 3.70(t, 2H, J=11Hz), 3.52~3.35(m, 6H), 3.02(m, 2H), 2.03~1.95(m, 2H ), 1.91 ~ 1.79 (m, 3H), 1.62 ~ 1.55 (m, 1H), 1.42 (t, 6H, J = 7.2Hz)

[234]

[235] Example 19: Preparation of N-(2-(1-pyrrolidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[236]

[237] The title compound was obtained by the same procedures as in Example 1, using 1-(2-aminoethyl)pyrrolidine instead of N-methylpiperazine (yield: 20.9%).

[238] TLC R _f = 0.08 in 10% MeOH in chloroform.

[239] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.69 (d, 1H, J = 4.0 Hz), 7.32 (d, 1H, J = 4.0 Hz), 7, 23 (d, 1H, J = 2.0 Hz), 7.17 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 6.86 (d, 1H, J = 8.0 Hz) , 3.94(s, 3H), 3.76(t, 2H, J=6.0Hz), 3.85~3.78(m, 2H), 3.75(t, 2H, J=6 .0Hz), 3.46(t, 2H, J=6.0Hz), 3.22~3.15(m, 2H), 2.28~2.13(m, 2H), 2.13 ~ 2.00 (m, 2H)

[240]

[241] Example 20: Preparation of N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[242]

[243] The title compound was prepared by the same procedures as in Example 1 using 4-(aminomethyl)-N,N-dimethyltetrahydro-2H-pyran-4-amine instead of N-methylpiperazine (yield: 48.4% ).

[244] TLC R _f = 0.37 in 10% MeOH in chloroform.

[245] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.80 (d, 1H, J = 4.0 Hz), 7.34 (d, 1H, J = 4.0 Hz), 7, 23 (d, 1H, J = 2.0 Hz), 7.18 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 6.86 (d, 1H, J = 8.4 Hz) , 4.08 ~ 4.04 (m, 2H), 4.02 (s, 2H), 3.94 (s, 3H), 3.82 ~ 3.75 (m, 2H), 2.99 (s , 6H), 2.05 ~ 1.94 (m, 4H)

[246]

[247] Example 21: Preparation of N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[248]

[249] The title compound was prepared following the same procedures as in Example 13 using N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-hydroxy-3 -methoxyphenyl)thiophene-2-carboxamide obtained by the same procedures as in Example 20, instead of N-(2-(diethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide ( yield: 72.3%).

[250] TLC R _f = 0.44 in 20% MeOH in chloroform.

[251] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.87 (d, 1H, J = 4.0 Hz), 7.43 (d, 1H, J = 4.0 Hz), 7, 33 (d, 1H, J = 2.0 Hz), 7.31 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 7.12 (d, 1H, J = 8.4 Hz) , 4.42(t, 2H, J=4.8Hz), 4.08~4.02(m, 2H), 4.03(s, 2H), 3.97(s, 3H), 3, 82 ~ 3.75 (m, 2H), 3.66 (t, 2H, J = 4.8Hz), 2.99 (s, 6H), 2.04 ~ 1.94 (m, 4H), 1 .42 (t, 6H, J = 7.2 Hz)

[252]

[253] Example 22: Preparation of N-(4-(1-methyl)piperidinyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[254]

[255] The title compound was prepared by the same procedures as in Example 1 using 4-amino-1-methylpiperidine instead of N-methylpiperazine (yield: 63.4%).

[256] TLC R _f = 0.20 in 20% MeOH in chloroform.

[257] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.71 ~ 7.67 (m, 1H), 7.30 (d, 1H, J = 4.0 Hz), 7.22 (d , 1H, J = 2.0 Hz), 7.17 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 6.86 (d, 1H, J = 8.0 Hz), 4, 18 ~ 4.08 (m, 1H), 3.94 (s, 3H), 3.62 ~ 3.57 (m, 2H), 3.25 ~ 3.14 (m, 2H), 2.92 ( s, 3H), 2.31 ~ 2.24 (m, 2H), 1.98 ~ 1.87 (m, 2H)

[258]

[259] Example 23: Preparation of N-(2-(2-(1-methyl)pyrrolidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[260]

[261] The title compound was obtained by the same procedures as in Example 1, using 2-(2-aminoethyl)-1-methylpyrrolidine instead of N-methylpiperazine (yield: 55.6%).

[262] TLC R _f = 0.16 in 20% MeOH in chloroform.

[263] ¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.64 (d, 1H, J = 4.0 Hz), 7.31 (d, 1H, J = 4.0 Hz), 7.22 ( d, 1H, J = 2.0 Hz), 7.17 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 6.86 (d, 1H, J = 8.0 Hz), 3 .94 (s, 3H), 3.73 ~ 3.66 (m, 1H), 3.53 ~ 3.48 (m, 2H), 3.41 ~ 3.35 (m, 1H), 3.23 ~ 3.15 (m, 1H), 2.97 (s, 3H), 2.57 ~ 2.47 (m, 1H), 2.35 ~ 2.25 (m, 1H), 2.25 ~ 2 .04 (m, 2H), 1.92 ~ 1.80 (m, 2H)

[264]

[265] Example 24: Preparation of N-(2-(2-(1-methyl)pyrrolidino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[266]

[267] The title compound was prepared following the same procedures as in Example 13 using N-(2-(2-(1-methyl)pyrrolidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene -2-carboxamide obtained by the same procedures as in Example 23 instead of N-(2-(diethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide (yield: 60, 9 %).

[268] TLC R _f = 0.12 in 20% MeOH in chloroform.

[269] ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.37 (s, 1H), 10.21 (s, 1H), 8.71 (t, 1H, J = 6.0 Hz), 7.78 (d, 1H, J = 4.0 Hz), 7.51 (d, 1H, J = 4.0 Hz), 7.30 (d, 1H, J = 2.0 Hz), 7, 26 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 7.10 (d, 1H, J = 8.0 Hz), 4.39 (t, 2H, J = 4.8 Hz) , 3.88 (s, 3H), 3.48 ~ 3.51 (m, 3H), 3.43 ~ 3.15 (m, 5H), 3.10 ~ 3.00 (m, 1H), 2 .79 (d, 3H, J = 4.8Hz), 2.40 ~ 2.35 (m, 1H), 2.22 ~ 2.13 (m, 1H), 2.07 ~ 1.80 (m , 3H), 1.72~1.63(m, 1H), 1.27(t, 6H, J=7.2Hz)

[270]

[271] Example 25: Preparation of N-((S)-2-(1-ethyl)pyrrolidinomethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[272]

[273] The title compound was obtained by the same procedures as in Example 1 using (S)-2-(aminomethyl)-1-ethylpyrrolidine instead of N-methylpiperazine (yield: 42.1%).

[274] TLC R _f = 0.29 in 20% MeOH in chloroform.

[275] ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.85 (s, 1H), 9.44 (s, 1H), 8.97 (t, 1H, J = 5.6 Hz), 7.82 (d, 1H, J = 4.0 Hz), 7.44 (d, 1H, J = 4.0 Hz), 7.22 (d, 1H, J = 2.0 Hz), 7, 13(dd, 1H, J=2.0Hz, 8.0Hz), 6.83(d, 1H, J=8.0Hz), 3.85(s, 3H), 3.72~3, 65(m, 1H), 3.65~3.53(m, 2H), 3.48~3.38(m, 2H), 3.15~3.05(m, 2H), 2.18~ 2.08(m, 1H), 2.03~1.93(m, 1H), 1.93~1.76(m, 2H), 1.28(t, 3H, J=7.2Hz)

[276]

[277] Example 26: Preparation of N-((S)-2-(1-ethyl) pyrrolidinomethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride

[278]

[279] The title compound was prepared following the same procedures as in Example 13 using N-((S)-2-(1-ethyl)pyrrolidinomethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene- 2-carboxamide obtained by the same procedures as in Example 25 instead of N-(2-(diethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide (yield: 37.6 %).

[280] TLC R _f = 0.24 in 20% MeOH in chloroform.

[281] ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.41 (s, 1H), 10.25 (s, 1H), 9.14 (t, 1H, J = 5.6 Hz), 7.92 (d, 1H, J = 4.0 Hz), 7.53 (d, 1H, J = 4.0 Hz), 7.31 (d, 1H, J = 2.0 Hz), 7, 27 (dd, 1H, J = 2.0 Hz, 8.0 Hz), 7.11 (d, 1H, J = 8.0 Hz), 4.40 (t, 2H, J = 4.8 Hz) , 3.88 (s, 3H), 3.82 ~ 3.72 (m, 1H), 3.67 ~ 3.50 (m, 4H), 3.48 ~ 3.40 (m, 2H), 3 .40 ~ 3.18 (m, 4H), 3.18 ~ 3.03 (m, 2H), 2.18 ~ 2.08 (m, 1H), 2.03 ~ 1.93 (m, 1H) , 1.93 ~ 1.75 (m, 2H), 1.45 ~ 1.35 (m, 9H)

[282]

[283] Test Example 1: Autophagy Inducing Activity

[284] The autophagy-inducing activity of the present invention was measured in Hela cell lines (Korean Cell Line Bank) using an autophagy detection kit (ab139484, Abcam) according to the manufacturer's instructions. Specifically, 100 µl of EBSS (Earle's Balanced Salt Solution, WELGENE (LB 002-03)) containing 10% fetal bovine serum (FBS) was added to each well of a 96-well plate. Hela cell lines (1 x 104 cells) were added to each well and then incubated overnight at 37° C. in a CO ₂ incubator to stabilize the cells. The medium of each well containing stabilized cells was replaced with medium prepared by adding 10 nm or 100 nm of the compounds obtained in Examples (test compounds) to EBSS containing 10% FBS. As a positive control, rapamycin was dissolved in dimethyl sulfoxide and then treated at a concentration of 500 nm per well. Following treatments, cells were incubated at 37° C. in a CO ₂ incubator for 4 or 24 hours and then washed twice with 1X assay buffer (prepared by adding 9 ml of distilled water to 10X assay buffer). EBSS containing 5% fetal bovine serum, 1 μl/ml of Green's developing reagent and 1 μl/ml of nuclear stain were added thereto in an amount of 100 μl per well. Cells were incubated for 1 hour at 37° C. in a CO ₂ incubator and then washed twice with 1X assay buffer. Absorbance values were measured at 488 nm using a plate reader (Cytation 3). The tests were repeated four times.

[285] The absorbance values that were obtained by treating Hela cell lines with test compounds (100nm) and a positive control (rapamycin, 500nm) followed by a 4 hour incubation as described above are shown in Table 1 below. Also, absorbance values that were obtained by treating Hela cell lines with test compounds (10 nM or 100 nM) and a positive control (rapamycin, 500 nM) followed by a 24 hour incubation as described above are shown in Table 2 below.

[286] Table 1

Absorption rate (incubation for 4 hours)

Test formulations (concentration) Absorption coefficient (at 488 nm) Example 1 (100 nm) 1321.2 Example 2 (100 nm) 1460.5 Example 3 (100 nm) 1193.8 Example 4 (100 nm) 1580.6 Example 5 (100 nm) 1348.0 Example 6 (100 nm) 1670.5 Example 7 (100 nm) 1325.5 Example 8 (100 nm) 1315.2 Example 9 (100 nm) 1320.2 Example 10 (100 nm) 1390.0 Example 11 (100 nm) 1268.4 Rapamycin (500 nm) 1306.2

[287]

[288] Table 2

Absorption rate (incubation for 24 hours)

Test formulations Absorption coefficient (at 488 nm) 10 nm 100 nm Example 1 1617.3 1660.3 Example 2 1829.3 1866.3 Example 3 1576.3 1754.3 Example 4 1691.3 1618.3 Example 5 1557.3 1618.3 Example 6 1784.6 1893.7 Example 7 1598.5 1605.5 Example 8 1586.3 1612.3 Example 9 1594.0 1951.7 Example 10 1729.3 1882.3 Example 11 1459.7 1731.0 Rapamycin (500 nm) 1594.2

[289] From the results shown in Table 1 above, it can be seen that when incubated for 4 hours after treatment of the test compounds, the compounds of this invention showed autophagy-inducing activity even at a concentration of 1/5 equal to or higher than that of the positive control. (i.e. rapamycin). In particular, from the results shown in Table 2, it can be seen that when incubated for 24 hours after treatment of the test compounds, the compounds of the present invention showed excellent (i.e., at least five times or more) autophagy-inducing activity even at concentrations of 1/50 and 1/5 compared to the positive control (i.e. rapamycin). Therefore, the compounds of the present invention exhibit excellent autophagy-inducing activity, whereby they can be successfully used in the prevention, alleviation or treatment of various autophagy-related diseases, including neurodegenerative diseases, liver diseases, metabolic diseases, sepsis, and so on.

[290]

[291] Test Example 2: Evaluation of Oral Liver Function Improvement Activity in a Liver Injury Model (1)

[292] Compounds of this invention were orally administered to male SD rats with dimethylnitrosamine (DMN)-induced liver injury for 3 weeks to evaluate activity in improving liver function. Specifically, 7-week-old male SD rats (Orient Bio, Korea) were placed in a laboratory environment at room temperature for 7 days. General external manifestations were observed, after which only healthy animals were used for the experiment. The rats were divided into 9 groups (n = 5 for each group): a normal control group, a group administered with DMN alone, and groups administered as a compound of this invention (compounds of Examples 2, 5, 6, 9, 10, 20 or 21) and DMN. DMN was dissolved in purified water and then administered intraperitoneally at a dose of 10 mg/kg for 3 consecutive weeks (3 times during the 4th week). Blood sampling was done on Day 3 of the first week after completion of liver injury induction, after which ALT (alanine transaminase) and AST (aspartate transaminase) values were measured to confirm liver injury. Compounds according to this invention were administered within 3 weeks, ie after Day 4 after completion of the induction of liver damage during the first week in relation to the period of administration of DMN. The seven compounds of this invention were dissolved in purified water or corn oil and then administered orally by oral gavage at a dose of 25 mg/kg once a day for 3 weeks. Blood sampling was done on Day 0 (on the 3rd day of the first week after completion of the induction of liver damage) and on Days 7, 14 and 21 after the administration of the test compounds. The collected blood samples were introduced into a vacuum tube containing a clot activator and then kept at room temperature for about 20 minutes to coagulate each blood sample. After centrifugation for 10 minutes, the resulting sera were subjected to biochemical blood tests. The experimental methods are summarized in FIG. 1.

[293] the Values of serum ALT and AST obtained when conducting a biochemical analysis of blood, as described above, are presented in Tables 3 and 4 below.

[294] Table 3

Group ALT values (unit: U/L, mean value) 0 week 1 Week 2 weeks 3 weeks Normal control group 45.64 54.04 53.68 55.00 Group treated with DMN 72.58 107.52 124.82 156.26 Group administered Compound of Example 2 and DMN 67.24 101.70 113.22 126.10 Group administered Compound of Example 5 and DMN 68.90 96.60 108.80 111.00 Group administered Compound of Example 6 and DMN 68.02 80.46 98.94 109.80 Group administered Compound of Example 9 and DMN 67.64 103.78 111.80 117.92 Group administered Compound of Example 10 and DMN 67.46 85.06 102.94 102.94 Group administered Compound of Example 20 and DMN 69.80 87.26 104.34 116.14 Group administered Compound of Example 21 and DMN 70.98 98.84 104.54 101.18

[295]

[296] Table 4

Group AST values (unit: U/L, mean value) 0 week 1 Week 2 weeks 3 weeks Normal control group 109.64 142.88 137.60 142.74 Group treated with DMN 140.16 207.10 250.84 301.04 Group administered Compound of Example 2 and DMN 143.32 210.86 223.88 250.76 Group administered Compound of Example 5 and DMN 137.66 187.26 219.66 234.22 Group administered Compound of Example 6 and DMN 141.78 220.86 215.42 249.56 Group administered Compound of Example 9 and DMN 140.64 220.58 241.06 222.50 Group administered Compound of Example 10 and DMN 145.00 202.52 218.60 199.34 Group administered Compound of Example 20 and DMN 136.28 194.88 209.56 229.74 Group administered Compound of Example 21 and DMN 142.10 201.30 208.26 195.10

[297] As can be seen from the results shown in Tables 3 and 4, serum ALT and AST values in the DMN treated group increased by about 3-fold and about 2-fold after 3 weeks, respectively. However, in the groups administered both the compounds of this invention and DMN, the ALT values 3 weeks after administration were 101.18~126.10 U/L (i.e. decreased by about 19~35% compared to given values in the DMN administered group); and AST values 3 weeks after administration were 195.10 ~ 250.76 U/L (i.e. decreased by about 17 ~ 35% compared with those values in the DMN administered group). Thus, it can be confirmed that the compounds of the present invention exhibit effective liver improving activity in liver injury, including liver fibrosis.

[298]

[299] Test Example 3: Evaluation of Oral Liver Function Improvement Activity in a Liver Injury Model (2)

[300] Compounds of this invention were orally administered to male SD rats with dimethylnitrosamine (DMN)-induced liver injury for 4 weeks to evaluate activity in improving liver function. Specifically, 7-week-old male SD rats (Orient Bio, Korea) were placed in a laboratory environment at room temperature for 7 days. Common external signs were observed, after which only healthy animals were used for the experiment. The rats were divided into 3 groups (n=10 for each group): a normal control group, a group treated with DMN alone, and groups treated with both the compound of the invention (compound of Example 21) and DMN. DMN was dissolved in purified water and then administered intraperitoneally at a dose of 10 mg/kg for 3 consecutive weeks (3 times a week for 4 weeks). Blood sampling was done after completion of liver injury induction at the fourth week, after which ALT (alanine transaminase) and AST (aspartate transaminase) values were measured to confirm liver injury. The compound of this invention was administered for 4 weeks starting on Day 1 after the completion of the 4 week induction of liver injury. The compound of the present invention (compound of Example 21) was dissolved in purified water and then administered orally by oral gavage at a dose of 25 mg/kg once a day for 4 weeks. Blood sampling was done on Day 0 (1 day after the completion of the four week induction of liver injury) and Days 7, 14, 21 and 28 after the start of the administration of the test compounds. The collected blood samples were introduced into a vacuum tube containing a clot activator and then kept at room temperature for about 20 minutes to coagulate each blood sample. After centrifugation for 10 minutes, the resulting sera were subjected to biochemical blood tests. In addition, 24 hours after the last injection, an autopsy was performed for hepatectomy and fixation to prepare their tissue samples. After preparing slides with tissue samples, they were stained with hematoxylin and eosin (H&E) to microscopically observe damage to liver tissue and infiltration of inflammatory cells in liver tissues. In addition, these tissue samples were also stained with Masson's trichrome for microscopic observation of collagen fiber deposits in the liver tissues. The experimental methods are summarized in FIG. 2.

[301] the Values of serum ALT and AST obtained when conducting a biochemical analysis of blood, as described above, are presented in Tables 5 and 6 below.

[302] Table 5

Group ALT values (unit: U/L, mean value) 0 week 1 Week 2 weeks 3 weeks 4 weeks Normal control group 50.90 51.15 50.09 48.38 48.79 Group treated with DMN 254.33 111.02 64.26 63.65 65.60 Group administered Compound of Example 21 and DMN 198.02 92.71 57.61 44.48 48.33

[303]

[304] Table 6

Group AST values (unit: U/L, mean value) 0 week 1 Week 2 weeks 3 weeks 4 weeks Normal control group 103.83 99.78 93.96 117.32 117.91 Group treated with DMN 339.76 182.01 132.13 143.45 158.68 Group administered Compound of Example 21 and DMN 329.92 158.91 107.61 112.06 110.45

[305] As can be seen from the results shown in Tables 5 and 6, the values of serum ALT and AST in the DMN treated group increased by about 4-5 times and about 3.2 times, respectively, after 4 weeks. However, in the group treated with both the compound of this invention (compound of Example 21) and DMN, ALT values 3 and 4 weeks after administration were 44.48 and 48.33 U/L, respectively (i.e. decreased by 30% and 26% compared to the given values in the DMN treated group); and AST values 3 and 4 weeks after administration were 112.06 and 110.45 U/L, respectively (i.e., decreased by about 22% and 30% compared to those values in the DMN treated group). Thus, it can be confirmed that the compounds of the present invention exhibit effective liver improving activity in liver injury including liver fibrosis.

[306] In addition, the results obtained by hematoxylin and eosin (H&E) staining and Masson's trichrome staining as described above are shown in FIG. 3 and FIG. 4 respectively. According to the results shown in FIG. 3 measured the degree of damage to the liver tissue and the degree of infiltration of inflammatory cells in the liver tissues. Also, according to the results shown in FIG. 4, the degree of deposition of collagen fibers in the liver tissues was measured. The results are shown below in Table 7.

[308] As can be seen from the results shown in FIG. 3 and Table 7, liver tissue damage and inflammatory cell infiltration in liver tissues in the DMN-administered group increased about 17-fold and about 5-fold after 4 weeks, respectively, compared to those in the normal control group. However, in the group administered both the compound of the present invention (compound of Example 21) and DMN, liver tissue damage and inflammatory cell infiltration in liver tissues were 276.0 cells/1000 cells and 85.33 cells/mm2, respectively. (i.e. an improvement of about 50% and 55% compared to the given values in the DMN treated group).

[309] Also, as can be seen from the results shown in FIG. 4 and Table 7, the deposits of collagen fibers in liver tissue in the DMN-administered group increased about 12-fold after 4 weeks compared to that in the normal control group. However, in the group administered both the compound of the present invention (Example 21 compound) and DMN, the collagen fiber deposition in the liver tissues was 10.98%/ ^mm value in the group administered with DMN).

[310] Thus, it can be confirmed that the compounds of the present invention exhibit effective liver improving activity in liver injury, including liver fibrosis.

Claims (105)

1. The compound of formula 1 or its pharmaceutically acceptable salt: Formula 1

wherein Y is O or S, (1) if Y is O, R ₁ is hydrogen or a C ₁ -C ₄ alkyl group substituted with mono- or di-C ₁ -C ₅ alkylamino, R ₂ - C ₁ -C ₆ alkyl group, R ₃ is hydrogen and R ₄ is (4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl group, C ₁ -C ₄ alkyl group substituted with mono- or di-C ₁ -C ₅ alkylamino, C ₁ -C ₄ alkyl group substituted with a nitrogen-containing cyclic ring selected from the group consisting of morpholine, piperidine or pyrrolidine, wherein the nitrogen-containing cyclic ring is optionally substituted with C ₁ -C ₄ alkyl, or a piperidinyl group optionally substituted with C ₁ -C ₄ alkyl, or R ₃ and R ₄ are connected to each other by the nitrogen atom to which they are attached to form a piperazine ring, the piperazine ring being optionally substituted with C ₁ -C ₄ alkyl, (2) if Y is S, R ₁ and R ₂ are independently hydrogen; C ₁ -C ₆ alkyl group or C ₁ -C ₄ alkyl group substituted with mono- or di-C ₁ -C ₅ alkylamino, R ₃ is hydrogen and R ₄ is a C ₁ -C ₄ alkyl group substituted with mono- or di-C ₁ -C ₅ alkylamino, or R ₃ and R ₄ are connected to each other by the nitrogen atom to which they are attached to form a piperazine ring, the piperazine ring being optionally substituted with C ₁ -C ₄ alkyl. 2. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein Y-O R ₁ is hydrogen or a diethylaminoethyl group, and R ₂ is a methyl group. 3. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein Y-O R ₁ - hydrogen or diethylaminoethyl group, R ₂ - methyl group, R ₃ is hydrogen, and R ₄ - (4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl group, dimethylaminoethyl group, diethylaminoethyl group, diisopropylaminoethyl group, morpholinoethyl group optionally substituted with C ₁ -C ₄ alkyl, piperidinoethyl group optionally substituted with C ₁ -C ₄ alkyl, a pyrrolidinoethyl group optionally substituted with C ₁ -C ₄ alkyl, or a piperidinyl group optionally substituted with C ₁ -C ₄ alkyl. 4. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein Y-O R ₁ - hydrogen or diethylaminoethyl group, R ₂ - methyl group, R ₃ is hydrogen, and R ₄ - (4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl group. 5. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein Y-O R ₁ - hydrogen or diethylaminoethyl group, R ₂ is a methyl group, and R ₃ and R ₄ are connected to each other by the nitrogen atom to which they are attached to form a piperazine ring, optionally substituted with C ₁ -C ₄ alkyl. 6. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein Y-S, R ₁ is hydrogen or a diethylaminoethyl group, and R ₂ is a methyl group. 7. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein Y-S, R ₁ - hydrogen or diethylaminoethyl group, R ₂ - methyl group, R ₃ is hydrogen, and R ₄ is an isopropyl group or a diisopropylaminoethyl group. 8. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein Y-S, R ₁ - hydrogen or diethylaminoethyl group, R ₂ is a methyl group, and R ₃ and R ₄ are connected to each other by the nitrogen atom to which they are attached to form a piperazine ring, optionally substituted with C ₁ -C ₄ alkyl. 9. A compound or a pharmaceutically acceptable salt thereof, which is one or more compounds selected from the group consisting of: N-(4-methylpiperazino)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(4-methylpiperazino)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(4-methylpiperazino)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride, N-(4-methylpiperazino)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride, N-(2-(diisopropylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(diisopropylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(diisopropylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride, N-(2-(diisopropylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride, N-isopropyl-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-isopropyl-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-thiocarboxamide hydrochloride, N-isopropyl-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-thiocarboxamide, N-(2-(diethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(diethylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(dimethylamino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(dimethylamino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(4-morpholino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(1-piperidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(1-piperidino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(1-pyrrolidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(4-(1-methyl)piperidinyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(2-(1-methyl)pyrrolidino)ethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-(2-(2-(1-methyl)pyrrolidino)ethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride, N-((S)-2-(1-ethyl)pyrrolidinomethyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride and N-((S)-2-(1-ethyl)pyrrolidinomethyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide hydrochloride. 10. N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-hydroxy-3-methoxyphenyl)thiophene-2-carboxamide or a pharmaceutically acceptable salt thereof. 11. N-((4-(dimethylamino)tetrahydro-2H-pyran-4-yl)methyl)-5-(4-(2-diethylamino)ethoxy-3-methoxyphenyl)thiophene-2-carboxamide or a pharmaceutically acceptable salt thereof . 12. A method for preparing a compound of Formula 1a or a pharmaceutically acceptable salt thereof, comprising coupling a compound of Formula 2 with a compound of Formula 3 to obtain a compound of Formula 1a: Formula 1a:

Formula 2:

Formula 3:

moreover, R ₁ , R ₂ , R ₃ and R ₄ are the same as defined in paragraph 1, Z is halogen, and R ₆ and R ₇ are hydrogen or are connected to each other by the boron atom to which they are attached to form 4,4,5,5-tetramethyl-1,3,2-dioxaborolane. 13. A method for preparing a compound of Formula 1b or a pharmaceutically acceptable salt thereof, comprising coupling a compound of Formula 6 with a compound of Formula 3 to obtain a compound of Formula 1b: Formula 1b:

Formula 6:

Formula 3:

moreover, R ₁ , R ₂ , R ₃ and R ₄ are the same as defined in paragraph 1, Z is halogen, and R ₆ and R ₇ are hydrogen or are connected to each other by the boron atom to which they are attached to form 4,4,5,5-tetramethyl-1,3,2-dioxaborolane. 14. The method according to p. 13, and the compound of Formula 6 is obtained by performing thioamidation of the compound of Formula 2: Formula 2:

moreover, R ₃ and R ₄ are the same as defined in paragraph 1, and Z is halogen. 15. A process for preparing a compound of Formula 1b or a pharmaceutically acceptable salt thereof, comprising performing a thioamidation of a compound of Formula 1a. Formula 1a:

Formula 1b:

and R ₁ , R ₂ , R ₃ , and R ₄ are the same as defined in paragraph 1. 16. The use of a compound or a pharmaceutically acceptable salt thereof according to one of paragraphs. 1-11 for preventing, alleviating, or treating neurodegenerative diseases selected from the group consisting of Huntington's disease, Parkinson's disease, Alzheimer's disease, prion disease, multiple sclerosis, and Lou Gehrig's disease; liver diseases selected from the group consisting of liver fibrosis, liver cirrhosis, hepatitis, and fatty liver disease; metabolic diseases selected from the group consisting of diabetes, hyperlipidemia, obesity and inflammation, or sepsis, by inducing autophagy.

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