Novel thiophene derivative and use thereof

Abstract

The present invention provides a novel thiophene derivative, an optical isomer thereof, or a pharmaceutically acceptable salt thereof. A novel thiophene derivative according to the present invention has excellent inhibitory activity against KDM4, and thus is useful as a therapeutic agent for brain diseases such as Alzheimer's disease.

Description

Novel thiophene derivatives and their uses

[0001] The present invention relates to a novel thiophene derivative, a pharmaceutical composition comprising the thiophene derivative, and a use of the compound.

[0002] Alzheimer's disease (AD) is a representative progressive neurodegenerative disease characterized by the degeneration of specific brain regions, leading to memory loss and cognitive impairment. Pathological hallmarks include amyloid plaques caused by beta-amyloid (Aβ) deposition, neurofibrillary tangles caused by phosphorylated tau protein deposition, and damaged neurons. Recent research on AD has identified various therapeutic targets, but relevant treatments have not yet been developed sufficiently.

[0003] Meanwhile, the blood-brain barrier (BBB) ​​is primarily composed of endothelial cells, pericytes, and astrocytes, protecting central neurons by restricting the passage of molecules and blood cells. It has been reported that blood-brain barrier dysfunction caused by persistent vascular inflammation can increase vascular permeability for various substances and blood cells, alter molecular signals, and affect the brain, thereby inducing and promoting the progression of brain diseases. However, considering factors such as drug permeation, the BBB is difficult to utilize as a therapeutic target. Therefore, development is still primarily focused on drugs aimed at simple permeation, and its use for therapeutic purposes is extremely limited.

[0004] As above, the mechanisms related to neuronal cell death, synapse loss, and neurofibrillary tangle formation have not been clearly identified to date, making it difficult to prevent and treat Alzheimer's disease and related dementia.

[0005] Against this backdrop, cytotoxicity of cholinergic neurons due to beta-amyloid aggregation is known to be the main cause of Alzheimer's disease, and many research and development efforts have been conducted in this regard. That is, various approaches to treating Alzheimer's disease targeting beta-amyloid, such as inhibiting beta-amyloid production (inhibiting β-secretase and γ-secretase), suppressing reactive oxygen species or inflammation caused by beta-amyloid, or removing beta-amyloid in the brain using amyloid antibodies, are being developed. However, the results related to these approaches have not yet shown sufficient results from a therapeutic perspective.

[0006] Meanwhile, leukocyte infiltration, driven by cell adhesion molecules (CAMs), is mediated by CAMs such as ICAM1 and VCAM1. While these proteins are normally under low expression, their expression increases dramatically in chronic diseases such as vascular inflammation, hypertension, and diabetes. This increase in CAMs leads to increased leukocyte migration into the brain, inducing brain inflammation. Many degenerative brain diseases are reported to be accompanied by brain inflammation. For example, increased levels of CAMs have been reported in the plasma of Alzheimer's disease patients. Therefore, suppressing CAM expression is expected to be an effective way to reduce brain inflammation.

[0007] Accordingly, the present inventors, while striving to find a new therapeutic agent to prevent or treat degenerative brain diseases including Alzheimer's disease and related dementias, discovered a novel histone lysine demethylase 4 (KDM4) inhibitor.

[0008] The present invention was completed by confirming that these KDM4 inhibitors can be used as new Alzheimer's treatment agents by improving cognitive function impairment, reducing beta-amyloid deposits, and effectively reducing brain inflammation.

[0009] The present invention provides a novel thiophene derivative, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[0010] The terminology used in this application is merely used to describe specific embodiments and is not intended to limit the present invention. The singular expression includes the plural expression unless the context clearly indicates otherwise. In this application, terms such as "comprise" or "have" should be understood to indicate the presence of a feature, step, structure, or combination thereof described above, but do not preclude the presence or addition of one or more other features, steps, structures, or combinations thereof.

[0011] The meanings of terms and symbols used in this document are as follows.

[0012] In the present invention, the term “halogen” means a substituent selected from fluorine (F), chloro (Cl), bromo (Br), and iodo (I).

[0013] In the present invention, the term “unsubstituted” means a state in which no substituent is substituted and is absent or hydrogen.

[0014] In the present invention, the term "substituted" refers to a moiety having a substituent that replaces a hydrogen atom on one or more carbon atoms of the main chain. "Substituted" or "substituted with" is defined to include the implicit condition that such substitution results in a stable compound, for example, a compound that does not undergo spontaneous transformation by rearrangement, cyclization, elimination, etc., depending on the permissibility of the substituted atom and the substituent.

[0015] In the present invention, "single bond" means that two connected radicals are directly connected. For example, if L represents a single bond in ALZ, this structure is essentially AZ.

[0016] In the present invention, "C x-y " means having carbon number x or more and y or less.

[0017] As used herein, the term “C1-C5 alkyl” refers to a straight or branched C1-C5 saturated hydrocarbon group, such as, for example, methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, etc. Preferred alkyl groups contain about 1, 2, 3, 4, or 5 carbon atoms in the chain. A branched chain means that one or more lower alkyl groups, for example, methyl, ethyl, or propyl, are attached to a linear alkyl chain. “Lower alkyl” refers to a group having about 1 to about 5 carbon atoms in the chain, which may be straight or branched. An “alkyl” may be unsubstituted or optionally substituted by one or more substituents, which may be the same or different. Each substituent can be halogen, alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, cyano, hydroxy, alkoxy, alkylthio, amino, carboxy, etc. Each of these substituents can follow any of the definitions for each substituent mentioned in the present specification.

[0018] In the present invention, the term “C1-C5 haloalkyl” means a C1-C5 straight-chain or branched-chain saturated hydrocarbon in which at least one hydrogen atom is replaced with a halogen atom (i.e., F, Cl, Br, or I). For example, and not limited thereto, it may be CH2F, CHF2, CF3, etc.

[0019] In the present invention, the term "C6-C 12"Aryl" refers to an aromatic hydrocarbon containing 6 to 12 carbon atoms. For example, it can refer to a ring system such as monocyclic (e.g., phenyl); bicyclic (e.g., indenyl, naphthalenyl, tetrahydronaphthyl, tetrahydroindenyl). Preferably, aryl can be a phenyl group having a chemical formula of C6H5 and in which 6 carbon atoms are arranged in a cyclic ring structure. The phenyl group is very stable and is a type of aromatic hydrocarbon found in many organic compounds. In addition, the aryl can be substituted or unsubstituted, and when substituted, the hydrogen at the ortho, meta, or para position of the phenyl ring is substituted with halogen, C1-C5 alkyl, C1-C5 haloalkyl, -O-C1-C5 haloalkyl, -O-C1-C5 alkyl, -C(=O)-(C1-C5 alkyl), -C(=O)-O-(C1-C5 alkyl), -OH, -NO2 or -NH2 may be substituted.

[0020] In the present invention, the term "C2-C 12"Heteroaryl" refers to an optionally substituted aromatic ring containing 2 to 12 carbon atoms, wherein at least one of the ring carbon atoms is replaced by a heteroatom selected from oxygen (O), nitrogen (N) and sulfur (S), or an aromatic ring (e.g., a bicyclic or tricyclic ring system) fused to one or more rings such as a heteroaryl ring, an aryl ring, a heterocyclic ring, or a carbocyclic ring, each of which may have an optional substituent. Examples thereof include pyrrole, pyrazole, imidazole, triazole (e.g., 1,2,3-triazole, 1,2,4-triazole, 1,2,4-triazole), tetrazole, furan, isoxazole, oxazole, oxadiazole (e.g., 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,3,4-oxadiazole), thiophene, isothiazole, thiazole, thiadiazole (e.g., 1,2,3-thiadiazole, 1,2,4-thiadiazole, 1,3,4-thiadiazole), pyridine, pyridazine, pyrimidine, pyrazine, triazine (e.g., 1,2,4-triazine, 1,3,5-triazine) and tetrazine, and may include monocyclic heteroaryls including indole, isoindole, indazole, benzimidazole, benzotriazole, benzopyrrole, benzofuran, benzoxazole, benzoisoxazole, benzoxadiazole, benzothiophene, benzothiazole, benzoisothiazole, benzothiadiazole, 1H-Pyrrolo[2,3-b]pyridine, 1H-pyrazolo[3,4-b]pyridine, 3H-imidazo[4,5-b]pyridine, 3H-[1,2,3]triazolo[4,5-b]pyridine, 1H-pyrrolo[3,2-b]pyridine, 1H-pyrazolo[4,3-b]pyridine, 1H-imidazo[4,5-b]pyridine, 1H-[1,2,3]triazolo[4,5-b]pyridine, 1H-pyrrolo[2,3-c]pyridine, 1H-pyrazolo[3,4-c]pyridine, 3H-imidazo[4,5-c]pyridine, 3H-[1,2,3]triazolo[4,5-c]pyridine, 1H-pyrrolo[3,2-c]pyridine, 1H-pyrazolo[4,3-c]pyridine, 1H-imidazo[4,5-c]pyridine, 1H-[1,2,3]triazolo[4,5-c]pyridine, furo[2,3-b]pyridine, oxazolo[5,4-b]pyridine, isoxazolo[5,4-b]pyridine, [1,2,3]oxadiazolo[5,4-b]pyridine, furo[3,2-b]pyridine, oxazolo[4,5-b]pyridine, isoxazolo[4,5-b]pyridine, [1,2,3]oxadiazolo[4,5-b]pyridine, furo[2,3-c]pyridine, oxazolo[5,4-c]pyridine, isoxazolo[5,4-c]pyridine, [1,2,3]oxadiazolo[5,4-c]pyridine, furo[3,2-c]pyridine, oxazolo[4,5-c]pyridine, Isoxazolo[4,5-c]pyridine, [1,2,3]oxadiazolo[4,5-c]pyridine, thieno[2,3-b]pyridine, thiazolo[5,4-b]pyridine, isothiazolo[5,4-b]pyridine, [1,2,3]thiadiazolo[5,4-b]pyridine, thieno[3,2-b]pyridine, thiazolo[4,5-b]pyridine, isothiazolo[4,5-b]pyridine, [1,2,3]thiadiazolo[4,5-b]pyridine, thieno[2,3-c]pyridine, thiazolo[5,4-c]pyridine, isothiazolo[5,4-c]pyridine, [1,2,3]thiadiazolo[5,4-c]pyridine, thieno[3,2-c]pyridine, thiazolo[4,5-c]pyridine, isothiazolo[4,5-c]pyridine, [1,2,3]thiadiazolo[4,5-c]pyridine, quinoline, isoquinoline, cinnoline, quinazoline, quinoxaline, phthalazine, naphthyridine (e.g., 1,8-naphthyridine, 1,7-naphthyridine, 1,6-naphthyridine, 1,5-naphthyridine, 2,7-naphthyridine, 2,6-naphthyridine), imidazo[1,2-a]pyridine, 1H-pyrazolo[3,4-d]thiazole, It may include polycyclic heteroaryl such as 1H-pyrazolo[4,3-d]thiazole and imidazo[2,1-b]thiazole, but is not limited thereto. In addition, the heteroaryl may be substituted or unsubstituted, and when substituted, it may be substituted with halogen, C1-C5 alkyl, C1-C5 haloalkyl, -O-C1-C5 haloalkyl, -O-C1-C5 alkyl, -C(=O)-(C1-C5 alkyl), -C(=O)-O-(C1-C5 alkyl), -OH, -NO2 or -NH2. In addition, the polycyclic heteroaryl group is,If the polycyclic heteroaryl group is attached to the parent structure via an aromatic ring, it may include a non-aromatic ring (e.g., cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl) fused to the heteroaryl ring.

[0021] In the present invention, “C2-C8 heterocycloalkyl” includes a saturated monocyclic or polycyclic heterocyclic ring containing 1 to 4 heteroatoms independently selected from nitrogen (N), oxygen (O), and sulfur (S), or a ring structure in which two or more rings share one or more pairs of carbon atoms (e.g., a fused ring, a spiro ring, a bridged ring, etc.). Heterocycloalkyl is azetidinyl, oxiranyl, oxetanyl, morpholinyl, thiomorpholinyl, furyl, piperazinyl, pyranyl, 1,3-dioxanyl, 2-oxopyrrolidinyl, 2-oxopiperidinyl, thietanyl, pyrrolidinyl, piperidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, imidazolidinyl, thiazolidinyl, azepanyl, diazepanyl, oxazepanyl, thiazepanyl, hexahydropyrrolo[1,2-a]pyrazin-6(2H)-one-yl, tetrahydro-1H-oxazolo[3,4-a]pyrazin-3(5H)-one-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-yl, 3-oxa-8-azabicyclo[3.2.1]octan-yl, hexahydro-2H-furo[3,2-b]pyrrole, azetidin-3-one, etc., but are not limited thereto. In addition, the heterocycloalkyl may be substituted or unsubstituted, and when substituted, may be substituted with halogen, C1-C5 alkyl, C1-C5 haloalkyl, -O-C1-C5 haloalkyl, -O-C1-C5 alkyl, -C(=O)-(C1-C5 alkyl), -C(=O)-O-(C1-C5 alkyl), -OH, -NO2 or -NH2. When nitrogen is present in a heterocycloalkyl ring, it may exist in an oxidized state (i.e., N+-O-) as long as the properties of adjacent atoms and groups permit. Examples include piperidinyl N-oxide and morpholinyl N-oxide. Furthermore, when sulfur is present in a heterocycloalkyl ring, it may exist in an oxidized state (i.e., S+-O- or -SO2-) as long as the properties of adjacent atoms and groups permit.Examples include thiomorpholine S-oxide and thiomorpholine S,S-dioxide. Additionally, one ring of the polycyclic heterocycloalkyl group may be aromatic (e.g., aryl or heteroaryl) if the polycyclic heterocycloalkyl group is attached to the parent structure via a non-aromatic carbon or nitrogen atom.

[0022] In the present invention, “enantiomer” refers to a case where two molecules having optical activity form a mirror-symmetric relationship. It is generally used as a synonym for mirror image isomer, and includes the R-form, S-form, or racemic compound forms, respectively.

[0023] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and will not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0024] Novel thiophene derivatives, optical isomers thereof, or pharmaceutically acceptable salts thereof

[0025] To solve the above-mentioned technical problem, the present invention provides a compound represented by the following chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof:

[0026] [Chemical Formula 1]

[0027]

[0028] In the above chemical formula 1,

[0029] R1 and R2 may be the same or different, and each independently represent unsubstituted or substituted C 1-5 It is alkyl;

[0030] n is an integer from 2 to 10;

[0031] R3 is -NRaRb, NH-Rc, or unsubstituted or substituted C containing at least one N. 2-8 is heterocycloalkyl;

[0032] Ra and Rb may be the same or different, and each independently represents -H or unsubstituted or substituted C. 1-5 It is alkyl;

[0033] Rc is unsubstituted or substituted C 1-5 Alkyl, unsubstituted or substituted C 6-12 Aryl, unsubstituted or substituted -(C 1-5 alkyl)-(C 6-12 Aryl), unsubstituted or substituted C 2-12 Heteroaryl, unsubstituted or substituted -(C 1-5 alkyl)-(C 2-12 heteroaryl), unsubstituted or substituted C 2-8 Heterocycloalkyl, or unsubstituted or substituted -(C 1-5 alkyl)-(C 2-8 heterocycloalkyl);

[0034] R4 is halogen, C 1-3 Alkyl, -O-(C 1-3 alkyl), -CN, -OH, -NH2 or -NO2.

[0035] Specifically, C1-C5 alkyl, C mentioned in the above R1 to R4 and Ra to Rc 6-12 Aryl, C 2-12 Heteroaryl, C2-C8 heterocycloalkyl can be selected from the specific substituents mentioned below:

[0036] C above 1-6 Alkyl may be straight or branched chain and may be methyl, ethyl, propyl, butyl, pentyl or hexyl.

[0037]

[0038] C above 6-12 Aryl is phenyl, the above C 6-12Aryl can be phenyl, naphthalenyl, indenyl, pentarenyl or azulenyl.

[0039] C above 2-12 Heteroaryl can be thiophenyl, furanyl, thiazolyl, oxazolyl, thiadiazolyl, oxadiazolyl, pyranyl, pyrazolyl, imidazolyl, triazolyl, pyridinyl, pyridonyl, pyrimidinyl, benzopyrrolyl, benzofuranyl, benzothiophenyl, isobenzofuranyl, benzoimidazolyl, benzothiazolyl, benzoisothiazolyl, benzoisooxazolyl, benzoxazolyl, isoindolyl, indolyl, indazolyl, benzothiadiazolyl, quinolyl, isoquinolyl, cinnolinyl, quinazolinyl, quinoxalinyl, carbazolyl, or benzodioxozolyl.

[0040] C above 2-8 Heterocycloalkyl can be aziridinyl, azetidinyl, morpholinyl, thiomorpholinyl, piperazinyl, 1,3-dioxanyl, thietanyl, pyrrolidinyl, piperidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, imidazolidinyl, thiazolidinyl, azepanyl, diazepanyl, oxazepanyl, thiazepanyl, hexahydropyrrolo[1,2-a]pyrazin-6(2H)-one-yl, 3-oxa-8-azabicyclo[3.2.1]octan-yl or hexahydro-2H-furo[3,2-b]pyrrole.

[0041] Specifically, R1 and R2 may be the same or different from each other, and each may independently be any one selected from the group consisting of methyl, ethyl, propyl, and isopropyl, and more specifically, R1 and R2 may be the same or different from each other, and each may independently be methyl or ethyl.

[0042] Specifically, the R3 is any one selected from the group consisting of -NRaRb, NH-Rc, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, imidazolidinyl, thiazolidinyl, tetrahydrofuranyl, azepanyl, diazepanyl, oxazepanyl and thiazepanyl, wherein the R3 may be unsubstituted or substituted with at least one or more halogen, C1-C5 alkyl, C1-C5 haloalkyl, -O-C1-C5 haloalkyl, -O-C1-C5 alkyl, -C(=O)-(C1-C5 alkyl), -C(=O)-O-(C1-C5 alkyl), -OH, -NO2 or -NH2.

[0043] Also specifically, R3 is any one selected from the group consisting of -NRaRb, NH-Rc, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, imidazolidinyl, thiazolidinyl, tetrahydrofuranyl, azepanyl, diazepanyl, oxazepanyl and thiazepanyl,

[0044] Ra and Rb may be the same or different and are each independently methyl, ethyl, propyl or isopropyl,

[0045] Rc is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, phenyl, -(C 1-5 Alkyl)-phenyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, furanyl, isoxazolyl, oxazolyl, oxadiazolyl, thiophenyl, isothiazolyl, thiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, -(C 1-5 alkyl)-pyrrolyl, -(C 1-5 Alkyl)-pyrazolyl, -(C 1-5 alkyl)-imidazolyl, -(C 1-5 Alkyl)-triazolyl, -(C 1-5 alkyl)-furanyl, -(C 1-5 Alkyl)-isoxazolyl, -(C 1-5 alkyl)-oxazolyl, -(C 1-5Alkyl)-oxadiazolyl, -(C 1-5 alkyl)-thiophenyl, -(C 1-5 Alkyl)-isothiazolyl, -(C 1-5 Alkyl)-thiazolyl, -(C 1-5 Alkyl)-thiadiazolyl, -(C 1-5 alkyl)-pyridinyl, -(C 1-5 alkyl)-pyridazinyl, -(C 1-5 alkyl)-pyrimidinyl, -(C 1-5 alkyl)-pyrazinyl, -(C 1-5 Alkyl)-triazinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, imidazolidinyl, thiazolidinyl, tetrahydrofuranyl, azepanyl, diazepanyl, oxazepanyl, thiazepanyl, -(C 1-5 alkyl)-azetidinyl, -(C 1-5 alkyl)-pyrrolidinyl, -(C 1-5 alkyl)-piperidinyl, -(C 1-5 alkyl)-piperazinyl, -(C 1-5 Alkyl)-morpholinyl, -(C 1-5 alkyl)-thiomorpholinyl, -(C 1-5 alkyl)-imidazolidinyl, -(C 1-5 alkyl)-thiazolidinyl, -(C 1-5 alkyl)-tetrahydrofuranyl, -(C 1-5 Alkyl)-azepanyl, -(C 1-5 alkyl)-diazepanyl, -(C 1-5 Alkyl)-oxazepanyl and -(C 1-5 Any one selected from the group consisting of alkyl)-thiazepanyl,

[0046] Here, R3 may be unsubstituted or substituted with at least one or more halogen, C1-C5 alkyl, C1-C5 haloalkyl, -O-C1-C5 haloalkyl, -O-C1-C5 alkyl, -C(=O)-(C1-C5 alkyl), -C(=O)-O-(C1-C5 alkyl), -OH, -NO2 or -NH2.

[0047] More specifically, the above R3 is , , , , , , , , , , , , and is one selected from the group consisting of,

[0048] The above R3 may be unsubstituted or substituted with at least one or more halogen, C1-C5 alkyl, C1-C5 haloalkyl, -O-C1-C5 haloalkyl, -O-C1-C5 alkyl, -C(=O)-(C1-C5 alkyl), -C(=O)-O-(C1-C5 alkyl), -OH, -NO2 or -NH2.

[0049] More specifically, the above R3 is , , , , , , , , , , , , , , , , and It can be any one selected from the group consisting of .

[0050] Additionally, the above n is an integer of 2, 3, 4, 5, 6, 7, 8, 9, or 10. More preferably, it may be an integer of 2 to 5.

[0051] Also, the above R4 is -O-(C 1-3 alkyl) or -OH.

[0052] The compound represented by the above chemical formula 1 may be a compound represented by the following chemical formula 2-1 or 2-2:

[0053] [Chemical Formula 2-1]

[0054]

[0055] [Chemical Formula 2-2]

[0056]

[0057] In the above chemical formula 2-1 or 2-2, n, R1 to R4 and Ra to Rc are as described above in the above chemical formula 1.

[0058] According to a specific example of the present invention, the compound represented by the above chemical formula 1 may be any one selected from the group consisting of compounds described below.

[0059] N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide;

[0060] N-(3-(dimethylamino)propyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide;

[0061] N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxamide;

[0062] N-(3-(dimethylamino)propyl)-5-(3-hydroxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxamide;

[0063] N-(2-(dimethylamino)ethyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide;

[0064] N-(4-(dimethylamino)butyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide;

[0065] tert-Butyl 4-(4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoyl)piperazine-1-carboxylate;

[0066] N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(piperazine-1-carbonyl)phenyl)thiophene-2-carboxamide;

[0067] N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(4-methylpiperazine-1-carbonyl)phenyl)thiophene-2-carboxamide;

[0068] N-(3-(dimethylamino)propyl)-5-(4-(4-ethylpiperazine-1-carbonyl)-3-methoxyphenyl)thiophene-2-carboxamide;

[0069] N-(3-(dimethylamino)propyl)-5-(4-(4-isopropylpiperazine-1-carbonyl)-3-methoxyphenyl)thiophene-2-carboxamide;

[0070] N-(3-(dimethylamino)propyl)-5-(4-(ethylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide;

[0071] 5-(4-(diethylcarbamoyl)-3-methoxyphenyl)-N-(3-(dimethylamino)propyl)thiophene-2-carboxamide;

[0072] N-(3-(dimethylamino)propyl)-5-(4-(isopropylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide;

[0073] N-(3-(dimethylamino)propyl)-5-(4-(isobutylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide;

[0074] N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(piperidine-1-carbonyl)phenyl)thiophene-2-carboxamide;

[0075] N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(pyrrolidine-1-carbonyl)phenyl)thiophene-2-carboxamide;

[0076] 5-(4-(benzylcarbamoyl)-3-methoxyphenyl)-N-(3-(dimethylamino)propyl)thiophene-2-carboxamide;

[0077] N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(phenethylcarbamoyl)phenyl)thiophene-2-carboxamide;

[0078] N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(pyridin-3-ylcarbamoyl)phenyl)thiophene-2-carboxamide;

[0079] N-(3-(dimethylamino)propyl)-5-(4-((furan-2-ylmethyl)carbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide; and

[0080] N-(3-(dimethylamino)propyl)-5-(4-hydroxy-3-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide.

[0081]

[0082] In addition, the compound represented by chemical formula 1 of the present invention, an optical isomer thereof, or a pharmaceutically acceptable salt thereof can inhibit the activity of KDM4.

[0083] In addition, the compound represented by chemical formula 1 of the present invention, an optical isomer thereof, or a pharmaceutically acceptable salt thereof inhibits the expression of VCAM1.

[0084] In addition, the adhesion and infiltration of blood cells are inhibited through suppression of VCAM1 expression as described above.

[0085] It also effectively reduces brain inflammation, reduces the accumulation of beta-amyloid, and improves functional impairment.

[0086] Specifically, the compound represented by Chemical Formula 1 of the present invention, which is a KDM4 inhibitor, showed effects of reducing VCAM1 expression, reducing leukocyte adhesion, and reducing amyloid levels in 5xFAD mice, an Alzheimer's mouse model. Furthermore, it showed significant effects in improving cognitive function, and therefore, the compound represented by Chemical Formula 1 of the present invention has the potential to be developed as a therapeutic agent for Alzheimer's disease.

[0087] In the present invention, pharmaceutically acceptable salts refer to salts commonly used in the pharmaceutical industry, and include, for example, inorganic ionic salts manufactured with calcium, potassium, sodium, and magnesium; inorganic acid salts manufactured with hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid, and sulfuric acid; organic acid salts manufactured with acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, and hydroiodic acid; sulfonic acid salts manufactured with methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid; Amino acid salts made from glycine, arginine, lysine, etc.; and amine salts made from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc.; however, the types of salts meant in the present invention are not limited by these listed salts.

[0088] Pharmaceutical composition

[0089] In another aspect of the present invention, a pharmaceutical composition is provided comprising a compound represented by formula 1 as defined in any embodiment described herein, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[0090] In another aspect of the present invention, a pharmaceutical composition is provided comprising a compound represented by formula 1 as defined in any embodiment described herein, an optical isomer thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

[0091] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier, and may be formulated in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.

[0092] The pharmaceutically acceptable carriers include, but are not limited to, those commonly used in the art, such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition, the pharmaceutical composition of the present invention may include, but is not limited to, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants, and other pharmaceutically acceptable additives.

[0093] In another aspect of the present invention, a pharmaceutical composition for preventing or treating brain diseases is provided, comprising a compound represented by Chemical Formula 1 as defined in any embodiment described herein, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[0094] The above brain diseases include senile dementia including Alzheimer's disease, vascular dementia, mild cognitive impairment, Parkinson's disease, Huntington's disease, cerebral amyloid angiopathy, Down syndrome, stroke, systemic amyloid disease, amyotrophic lateral sclerosis, multiple sclerosis, stroke, encephalitis, spinocerebellar atrophy, Tourette's syndrome, Friedrich's ataxia, Lewy body dementia, progressive supranuclear palsy, frontotemporal dementia, and Wernicke-Korsakoff's disease. It may be any one selected from the group consisting of, but is not limited to, Wernicke-Korsakoff's syndrome.

[0095] Preferably, the brain disease may be Alzheimer's disease.

[0096] In the present invention, the term "Alzheimer's disease" is used interchangeably with senile dementia and may mean a disease accompanied by mental deterioration associated with a specific degenerative brain disease characterized by senile plaques, neuroinflammatory tangles, and progressive neuronal loss.

[0097] In the present invention, the term “prevention” means any act of suppressing or delaying the onset of a brain disease by administering a composition.

[0098] In the present invention, the term “treatment” means any action by which the symptoms of the brain disease are improved or beneficially changed by administration of the composition.

[0099] The pharmaceutical composition of the present invention can be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) depending on the intended method, and the dosage range varies depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and type and severity of the disease.

[0100] The route of administration of the pharmaceutical composition of the present invention may be administered to mammals such as rats, mice, livestock, and humans through any general route as long as it can reach the target tissue, but may be administered by subcutaneous injection using an osmotic pump, intradermal injection, intravein injection, intraperitoneal injection, intravitreal injection, intrathecal, inner ear, abdominal cavity, or intravenous, intramuscular, subcutaneous, intrauterine epidural, sublingual, or intracerebrovascular injection, but is not limited thereto.

[0101] The pharmaceutical composition of the present invention may contain 0.001 to 95 wt%, preferably 0.01 to 80 wt%, of the compound represented by Chemical Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, based on the total weight of the composition.

[0102] When the pharmaceutical composition of the present invention is formulated as an oral solid preparation, it includes tablets, pills, powders, granules, capsules, etc., and such solid preparations may include at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., and include, but are not limited to, lubricants such as magnesium stearate and talc.

[0103] When the pharmaceutical composition of the present invention is formulated as an oral liquid, it includes a suspension, a solution, an emulsion, a syrup, etc., and includes, but is not limited to, a diluent such as water or liquid paraffin, a wetting agent, a sweetener, a fragrance, a preservative, etc.

[0104] When the pharmaceutical composition of the present invention is formulated for parenteral use, it includes a sterile aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized preparation, and a suppository. Non-aqueous solvents and suspensions include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Bases for suppositories include, but are not limited to, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin.

[0105] In another aspect of the present invention, a KDM4 inhibitor is provided, comprising a compound represented by formula 1 as defined in any embodiment described herein, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[0106] In another aspect of the present invention, a health functional food composition for preventing or improving brain disease is provided, comprising a compound represented by Chemical Formula 1 as defined in any embodiment described herein, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[0107] The term "improvement" as used herein refers to any action that improves or benefits the symptoms of a subject suspected of having a brain disorder and the invention by using a composition comprising a compound represented by Formula 1 as defined in any embodiment described herein, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[0108] The term "functional food" used in the present invention is the same as food for special health use (FoSHU), and refers to a food with high medical and healthcare effects that is processed to efficiently exhibit a bioregulatory function in addition to providing nutrition. Here, "functionality" means obtaining a useful effect for health purposes, such as regulating nutrients for the structure and function of the human body or physiological action. The food of the present invention can be manufactured by a method commonly used in the art, and during the manufacturing process, it can be manufactured by adding raw materials and ingredients commonly added in the art.

[0109] In addition, the food formulation of the present invention can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in various forms of formulation, and unlike general drugs, it has the advantage of not having side effects that can occur with long-term use of drugs because it uses food as a raw material. In addition, since it is highly portable, the food of the present invention can be consumed as a supplement to enhance the effect of preventing or improving diseases.

[0110] The term "health food" refers to foods that have a more active health maintenance or promotion effect than regular foods, while "health supplement food" refers to foods intended for health supplementation. In some cases, the terms "health functional food," "health food," and "health supplement food" are used interchangeably.

[0111] The food composition of the present invention is very useful because it can be consumed on a daily basis and can be expected to have an excellent effect in preventing or improving brain diseases.

[0112] In one aspect of the present invention, a method of treating a brain disorder is provided, comprising administering to a subject in need thereof a therapeutically effective amount of a compound represented by Formula 1 as defined in any embodiment described herein, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[0113] In the present invention, the term “compound represented by chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof” or “brain disease” is as described above.

[0114] The term "subject" of the present invention refers to any animal that has developed or may develop a brain disease, and may typically be an animal that can exhibit a beneficial effect by treatment with a compound represented by Chemical Formula 1 of the present invention, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, but includes, without limitation, any subject that has symptoms of a brain disease or is likely to have such symptoms. As described above, by administering the pharmaceutical composition of the present invention to a subject, the above-described disease can be effectively prevented or treated. The pharmaceutical composition of the present invention can be administered as an individual therapeutic agent, or in combination with existing therapeutic agents for brain diseases, and can be administered sequentially or simultaneously with existing therapeutic agents.

[0115] The term "therapeutically effective amount" as used herein means an amount sufficient to prevent or treat a disease at a reasonable benefit / risk ratio applicable to medical prevention or treatment, and represents an amount of the compound represented by Chemical Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, which is effective in preventing or treating the disease. The effective dosage level may be determined according to factors including the severity of the disease, the activity of the drug, the age, weight, health, and sex of the patient, the patient's sensitivity to the drug, the time of administration, the route of administration, and the excretion rate of the composition of the present invention used, the duration of treatment, drugs combined with or used concurrently with the composition of the present invention used, and other factors well known in the medical field. For example, the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof may be administered at 0.0001 to 100 mg / kg per day, and the administration may be administered once a day or in several divided doses.

[0116] The term "administration" in the present invention means introducing a predetermined substance into a patient by an appropriate method, and the administration route of the composition may be administered through any common route as long as it can reach the target tissue. In addition, the pharmaceutical composition of the present invention may be administered by any device that allows the active substance to move to the target tissue. For example, it may be administered by oral administration, intrathecal administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, topical administration, intranasal administration, intrapulmonary administration, rectal administration, inner ear administration, intrauterine epidural administration, sublingual administration, and intracerebrovascular injection, but is not limited thereto. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories.

[0117] At this time, in order to formulate a dosage form for parenteral administration, the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof is mixed with a stabilizer or buffer in water to prepare a solution or suspension, which can be prepared in an ampoule or vial unit dosage form. The composition may be sterilized and / or contain auxiliary agents such as preservatives, stabilizers, wetting agents or emulsifying promoters, salts for osmotic pressure control and / or buffers, and other therapeutically useful substances, and may be formulated according to conventional mixing, granulation, or coating methods.

[0118] The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with another therapeutic agent, and may be administered sequentially or simultaneously with conventional therapeutic agents. Taking all of the above factors into account, the pharmaceutical composition may be administered in an amount that achieves maximum efficacy with minimal side effects, as can be readily determined by those skilled in the art.

[0119] The therapeutic method of the present invention includes not only treating the disease itself before the onset of symptoms, but also inhibiting or avoiding its symptoms by administering the compound represented by the above chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof. In the management of a disease, the prophylactic or therapeutic dosage of a particular active ingredient will vary depending on the nature and severity of the disease or condition, and the route by which the active ingredient is administered. The dosage and frequency of administration will vary depending on the age, weight, and response of the individual patient. An appropriate dosage regimen can be readily selected by one skilled in the art, taking these factors into account. In addition, the treatment method of the present invention may further include administration of a therapeutically effective amount of an additional active agent helpful in treating a disease together with the compound represented by the above chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the additional active agent may exhibit a synergistic or auxiliary effect together with the compound represented by the above chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[0120] In addition, the present invention provides the use of a compound represented by the above chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof for the manufacture of a drug for treating brain diseases.

[0121] In addition, the present invention provides a compound represented by the above chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof for the treatment of brain diseases.

[0122] In addition, the present invention provides a pharmaceutical composition comprising a compound represented by the above chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof for the treatment of brain diseases.

[0123] In addition, the present invention provides a use of a compound represented by the above chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof for the treatment of brain diseases.

[0124] The present invention is a novel thiophene derivative having excellent inhibitory activity against KDM4, and is therefore useful as a therapeutic agent for brain diseases such as Alzheimer's.

[0125] Figure 1 is a diagram showing the degree of KDM4C inhibition at a single concentration (1 μM) of compounds according to Examples 1 to 22. Statistical analysis was performed using one-way ANOVA (Dunnett's multiple comparisons test), and the results were expressed as mean ± SEM, and the significance notation is as follows (*, p < 0.05; **, p < 0.01, ***, p < 0.001).

[0126] Figure 2 is a drawing confirming the effect of the compound according to Example 1 on the adhesion protein increased by TNF-α treatment in HBMVEC (human brain microvascular endothelial cells). Bands were quantified using Image J to derive the results. Statistical analysis was performed using one-way ANOVA (Dunnett's multiple comparisons test), and the results were expressed as mean ± SEM. The significance notation is as follows (*, p < 0.05; **, p < 0.01, ***, p < 0.001).

[0127] Figure 3 is a drawing confirming the adhesion inhibition effect of HL-60 cells. After pretreatment of HBMVEC with the compound according to Example 1 for 1 hour, TNFα (10 ng / ml) was treated for 8 hours, and HL-60 cells (1x10) labeled with fluorescent 6) was added and cultured at 37°C for 1 hour and observed under a fluorescence microscope. The number of HL-60 cells treated with fluorescence was counted and statistically analyzed. Statistical analysis was performed using one-way ANOVA (Dunnett's multiple comparisons test), and the results were expressed as mean ± SEM (n = 5 to 8). Significance notation is as follows (*, p < 0.05; **, p < 0.01, ***, p < 0.001).

[0128] FIG. 4a is a diagram showing the administration schedule of the compound according to Example 1, and FIG. 4b is a diagram showing the body weight and feed intake measured during the administration period of the compound according to Example 1, dividing 5xFAD and WT mice into a group administered the compound (1 mg / kg) according to Example 1 and a non-administered group. In addition, FIGS. 4c to 4d are diagrams showing behavioral experiments conducted after administration of the compound according to Example 1. FIG. 4c is a diagram showing the open field test, FIG. 4d is a diagram showing the Y-maze, and FIG. 4e is a diagram showing the passive avoidance test, respectively. Statistical analysis was performed using one-way ANOVA (Dunnett's multiple comparisons test), and the results were expressed as mean ± SEM (n = 5 to 8). Significance notations are as follows (*, p < 0.05; **, p < 0.01, ***, p < 0.001).

[0129] Figure 5a is a diagram showing the results of DAB staining of amyloid plaques in the cortex (RSA and ECT) and hippocampus (CA1 and DG) of 5xFAD mice (Scale bar = 100 μm), and Figures 5b to 5e are diagrams showing the quantification of the area of ​​amyloid plaques (n = 4). Statistical analysis was performed using one-way ANOVA (Dunnett's multiple comparisons test), and the results are expressed as mean ± SEM (n = 5 to 8). Significance notation is as follows (*, p < 0.05; **, p < 0.01, ***, p < 0.001).

[0130] Hereinafter, the present invention will be described in detail using examples to aid understanding. However, the examples according to the present invention may be modified in various ways, and the scope of the present invention should not be construed as being limited to the following examples. The examples of the present invention are provided to more fully explain the present invention to those of average skill in the art.

[0131] The reagents and solvents mentioned below were purchased from Sigma-Aldrich, Alpha Aesar, TCI, Daejung Chemicals, and Samsung Pure & Metals, unless otherwise specified, and structural analysis and purification were performed under the following conditions.

[0132] ● HPLC analysis conditions

[0133] Device name: Waters Alliance 2685 and Waters 996 Photodiode Array Detector

[0134] Column: YMC-pack pro C18, 150x4.6mm ID, 5 νm, 40℃

[0135] Mobile phase: 25% -> 80% acetonitrile / H2O + 0.1% trifluoroacetic acid

[0136] Analysis time: 15 minutes, flow rate: 0.5 ml / min

[0137] UV detector: 254nm

[0138] ●LC-MS analysis conditions

[0139] Device Name: Waters micromass zq

[0140] Column: ACE Excel2 C18, 75x2.1 mm

[0141] Mobile phase: 55% acetonitrile / H2O + 0.1% formic acid

[0142] Flow rate: 0.5 mL / min

[0143] UV detector: 254nm

[0144] ● 1 H NMR & 13 C NMR analysis conditions

[0145] Device Name: Varian VNMRS500 (500 MHz & 126 MHz)

[0146] 1 H and 13 C nuclear magnetic resonance (NMR) spectra were all consistent with the chemical structures of the exemplary compounds of the present invention. Characteristic chemical shifts (δ) are given in parts-per-million (ppm) relative to residual proton signals in deuterated solvents (CDCl3: 7.26 ppm; CD3OD: 3.31 ppm; DMSO-d6: 2.50 ppm) and are reported by the conventional abbreviations for major peak designations: e.g., s, singlet; d, doublet; t, triplet; q, quadruplet; m, multiplet; br, broad.

[0147] <Example>

[0148] Examples 1 to 6 below were synthesized based on the following reaction scheme 1, and each substituent is specifically described in the reaction scheme. The reagents and conditions used in reaction scheme 1 are as follows. (a) Di-tert-butyldicarbonate, 4-DMAP, DIPEA, DCM, 55°C, overnight; (b) Boronic acid, Pd(PPh3)4, Cs2CO3, DME / Water (1:1), 68°C, overnight; (c) NaOH aqueous solution, MeOH, THF (1:1:3), 50°C, 30 min; (d) Amines, EDC·HCl, HOBt, TEA, DMF, rt, overnight; (e) TFA, DCM, rt, 2.5 hrs; (f) Aliphatic amine, EDC·HCl, HOBt, TEA, DMF, rt, overnight; (g) BBr3, DCM, -78℃→rt, 1.5hrs.

[0149] [Reaction Formula 1]

[0150]

[0151] Example 1. Preparation of N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide (7b)

[0152]

[0153] Step 1: Preparation of tert-butyl 5-bromothiophene-2-carboxylate

[0154] 5-Bromothiophene-2-carboxylic acid was dissolved in dichloromethane in an oven-dried, sealed tube. 4-(dimethylamino)pyridine and di-tert-butyl dicarbonate were added to this solution at room temperature. N,N-diisopropylethylamine was then added sequentially at the same temperature. The mixture was stirred overnight at 55°C. The mixture was cooled to room temperature, diluted with distilled water, and extracted three times with dichloromethane. The combined organic phases were dried over magnesium sulfate. The solvent was evaporated in vacuo. The residue was purified by flash column chromatography using a hexane-ethyl acetate (19:1) system, yielding a pale yellow oil.

[0155] Step 2: Preparation of tert-butyl 5-(4-methoxy-3-(methoxycarbonyl)phenyl)thiophene-2-carboxylate

[0156] The tert-butyl 5-bromothiophene-2-carboxylate obtained in Step 1 was added to a sealed, oven-dried tube at room temperature. (3-Methoxy-4-(methoxycarbonyl)phenyl)boronic acid and cesium carbonate were added at room temperature. Dimethyl ether and water were also added. The mixture was bubbled with an inert gas to remove dissolved oxygen. Tetrakis(triphenylphosphine)palladium was then added at room temperature. The mixture was heated to 68°C and stirred overnight. The reaction mixture was cooled to room temperature, diluted with distilled water, and extracted with ethyl acetate. The combined organic phases were dried over magnesium sulfate. The solvent was removed under vacuum. The residue was purified by flash column chromatography using a hexane-ethyl acetate (14:1) system to obtain a brown oil.

[0157] Step 3: Preparation of 4-(5-(tert-butoxycarbonyl)thiophene-2-yl)-2-methoxybenzoic acid

[0158] tert-Butyl 5-(4-methoxy-3-(methoxycarbonyl)phenyl)thiophene-2-carboxylate obtained in Step 2 was added to a mixture of tetrahydrofuran and methanol with 1 molar aqueous sodium hydroxide solution. The mixture was stirred at 50°C for 30 minutes. After confirming the reaction, the mixture was diluted with water and acidified with a 10% aqueous hydrogen chloride solution. It was then extracted three times with ethyl acetate. The combined organic phases were dried over magnesium sulfate. The solvent was removed under vacuum to give a white solid, which was used in the next reaction without further purification.

[0159] Step 4: Preparation of tert-butyl 5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxylate

[0160] 4-(5-(tert-butoxycarbonyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in Step 3 above was dissolved in N,N-dimethylformamide solvent, and then morpholine, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, hydroxybenzotriazole, and triethylamine were added at room temperature. The mixture was stirred at room temperature. After the reaction was completed, the reaction solution was diluted with dilute water and extracted three times with ethyl acetate. The organic solvent was washed with water and brine to remove N,N-dimethylformamide, and dried over magnesium sulfate. The solvent was removed in vacuo to give a brown oil, which was used in the next reaction without further purification.

[0161] Step 5: Preparation of 5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxylic acid

[0162] The tert-butyl 5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxylate obtained in Step 4 was dissolved in dichloromethane solvent at 0°C, and trifluoroacetic acid was added. The mixture was stirred at room temperature for 2.5 hours. After monitoring the disappearance of the reaction mass, the mixture was diluted with dilute water and acidified with a 10% aqueous hydrogen chloride solution. It was then extracted three times with dichloromethane. The organic solvent was dried over magnesium sulfate. The solvent was removed under vacuum to obtain a white solid, which was used in the next reaction without further purification.

[0163] Step 6: Preparation of N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide

[0164] 5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxylic acid obtained in Step 5 above was dissolved in N,N-dimethylformamide, and then N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, hydroxybenzotriazole, N,N-dimethyl-1,3-propanediamine, and triethylamine were added at room temperature. The mixture was stirred at room temperature overnight. After the reaction product appeared, it was diluted with distilled water and extracted. The organic solvent was washed with water and brine, and the organic phase was dried over magnesium sulfate to remove DMF. The solvent was removed under vacuum. Recrystallization afforded N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide as a brown solid (119 mg, 31.9% yield).

[0165] 1 H NMR (500 MHz, cd3od) δ 7.65 (d, J = 3.8 Hz, 1H), 7.48 (d, J = 3.8 Hz, 1H), 7.34 - 7.30 (m, 2H), 7.27 (d, J = 7.7 Hz, 1H), 3.92 (s, 3H), 3.81 - 3.68 (m, 4H), 3.66 - 3.53 (m, 2H), 3.40 (t, J = 6.9 Hz, 2H), 3.32 - 3.24 (m, 2H), 2.50 (t, 2H), 2.33 (s, 5H), 1.88 - 1.78 (m, 2H); 13C NMR (126 MHz, cd3od) δ 168.00, 162.60, 155.89, 147.53, 138.47, 136.32, 129.11, 128.55, 124.63, 124.58, 118.33, 108.21, 66.54, 66.25, 56.67, 54.99, 47.34, 43.83, 42.12, 37.66, 26.59; LC-MS [C 22 H 29 N3O4S] calcd for [M+H], 432.2 found 432.2.

[0166] Example 2. Preparation of N-(3-(dimethylamino)propyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide (8b)

[0167]

[0168] N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide obtained in step 6 of Example 1 was dissolved in dichloromethane, and 1 mol of boron tribromide was slowly added dropwise at -78 °C. The mixture was stirred at room temperature for 1.5 hours. The reaction was stopped using methanol, and the mixture was concentrated under reduced pressure. The above concentrate was purified by silica gel column chromatography to obtain N-(3-(dimethylamino)propyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide (53.1 mg, 95%).

[0169] 1H NMR (500 MHz, cd3od) δ 7.75 (d, J = 3.9 Hz, 1H), 7.41 (d, J = 3.8 Hz, 1H), 7.28 - 7.23 (m, 2H), 7.19 (s, 1H), 3.77 - 3.63 (m, 6H), 3.48 (t, J = 6.4 Hz, 3H), 3.34 (d, J = 10.9 Hz, 2H), 3.26 - 3.21 (m, 2H), 2.93 (s, 6H), 2.10 - 2.02 (m, 2H); LC-MS [C 21 H 27 N3O4S] calcd for [M+H], 418.2 found 418.2.

[0170] Example 3. Preparation of N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxamide (7d)

[0171]

[0172] Step 1: Preparation of tert-butyl 5-(3-methoxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxylate

[0173] 4-(5-(tert-butoxycarbonyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 3 of Example 1 was dissolved in N,N-dimethylformamide, and then aniline, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, hydroxybenzotriazole, and triethylamine were added at room temperature. The mixture was stirred at room temperature. After the reaction was completed, the reaction solution was diluted with dilute water and extracted three times with ethyl acetate. The organic solvent was washed with water and brine to remove N,N-dimethylformamide, and dried over magnesium sulfate. The solvent was removed in vacuo to give a brown oil, which was used in the next reaction without further purification.

[0174] Step 2: Preparation of N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxamide

[0175] Using tert-butyl 5-(3-methoxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxylate obtained in step 1 above, the final product N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxamide was obtained by performing similar methods and procedures as those used in steps 5 and 6 of Example 1 above. (92 mg, 68.2% yield)

[0176] 1H NMR (500 MHz, cdcl3) δ 9.76 (s, 1H), 8.66 (s, 1H), 8.28 (d, J = 8.1 Hz, 1H), 7.67 (d, J = 7.7 Hz, 2H), 7.45 (s, 1H), 7.39 - 7.31 (m, 3H), 7.21 (s, 1H), 7.12 (t, J = 7.0 Hz, 1H), 4.10 (s, 2H), 3.56 (d, J = 4.1 Hz, 2H), 2.64 - 2.57 (m, 2H), 2.39 (s, 5H), 1.87 - 1.77 (m, 2H); 13 C NMR (126 MHz, cd3od) δ 164.99, 163.41, 158.48, 147.86, 139.58, 138.95, 138.76, 132.38, 129.98, 129.33, 125.82, 124.96, 122.88, 121.16, 118.87, 109.34, 57.32, 56.26, 44.39, 38.31, 27.18; LC-MS [C 24 H 27 N3O3S] calcd for [M+H], 438.2 found 438.2.

[0177] Example 4. Preparation of N-(3-(dimethylamino)propyl)-5-(3-hydroxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxamide (8d)

[0178]

[0179] N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxamide obtained in Example 3 was dissolved in dichloromethane, and 1 mol of boron tribromide was slowly added dropwise at -78 ℃. The mixture was stirred at room temperature for 1.5 hours. The reaction was stopped using methanol, and the mixture was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain N-(3-(dimethylamino)propyl)-5-(3-hydroxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxamide (21 mg, 52.5% yield).

[0180] 1 H NMR (500 MHz, cd3od) δ 8.01 (d, J = 8.1 Hz, 1H), 7.69 (dd, J = 19.7, 5.5 Hz, 3H), 7.50 (d, J = 3.2 Hz, 1H), 7.37 (t, J = 7.5 Hz, 2H), 7.29 - 7.22 (m, 2H), 7.16 (t, J = 7.3 Hz, 1H), 3.47 (t, J = 6.2 Hz, 2H), 3.20 - 3.13 (m, 2H), 2.88 (s, 3H), 2.09 - 1.98 (m, 2H); 13 C NMR (126 MHz, cd3od) δ 166.79, 163.23, 159.53, 147.34, 138.41, 138.23, 137.71, 129.65, 129.45, 128.47, 124.97, 124.43, 121.13, 120.42, 116.51, 113.70, 55.33, 42.20, 36.08, 24.85; LC-MS [C 23 H 25 N3O3S] calcd for [M+H], 424.2 found 424.2.

[0181] Example 5. Preparation of N-(2-(dimethylamino)ethyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide (8a)

[0182]

[0183] Step 1: Preparation of N-(2-(dimethylamino)ethyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide

[0184] 5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxylic acid obtained in step 5 of Example 1 was dissolved in N,N-dimethylformamide, and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, hydroxybenzotriazole, N',N'-dimethylethane-1,2-diamine, and triethylamine were added at room temperature. The mixture was stirred at room temperature overnight. After the reaction product appeared, it was diluted with distilled water and extracted. The organic solvent was washed with water and brine, and dried over magnesium sulfate to remove DMF from the organic phase. The solvent was removed under vacuum. N-(2-(dimethylamino)ethyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide was obtained by recrystallization.

[0185] Step 2: Preparation of N-(2-(dimethylamino)ethyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide

[0186] By using the N-(2-(dimethylamino)ethyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide obtained in the above step 1, a method and procedure similar to that used in step 7 of the above example 1 were performed, thereby obtaining the final product, N-(2-(dimethylamino)ethyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide (40 mg, 83.3% yield).

[0187] 1 H NMR (500 MHz, dmso) δ 10.19 (s, 1H), 8.59 (t, J = 4.8 Hz, 1H), 7.74 (d, J = 3.8 Hz, 1H), 7.48 (d, J = 3.8 Hz, 1H), 7.21 (s, 2H), 7.14 (s, 1H), 3.59 (d, J = 1.3 Hz, 7H), 3.43 - 3.37 (m, 3H), 3.20 (d, J = 34.6 Hz, 5H), 2.64 (s, 2H), 2.37 (s, 6H); 13 C NMR (126 MHz, dmso) δ 166.93, 161.42, 154.22, 147.03, 139.49, 135.30, 129.73, 129.63, 125.04, 124.16, 117.12, 112.93, 66.63, 58.04, 44.93, 36.94; LC-MS [C 20 H 25N3O4S] calcd for [M+H], 404.2 found 404.2

[0188] Example 6. Preparation of N-(4-(dimethylamino)butyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide (8c)

[0189]

[0190] Step 1: Preparation of N-(4-(dimethylamino)butyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide

[0191] 5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxylic acid obtained in step 5 of Example 1 was dissolved in N,N-dimethylformamide, and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, hydroxybenzotriazole, N',N'-dimethylbutane-1,4-diamine, and triethylamine were added at room temperature. The mixture was stirred at room temperature overnight. After the reaction product appeared, it was diluted with distilled water and extracted. The organic solvent was washed with water and brine, and dried over magnesium sulfate to remove DMF from the organic phase. The solvent was removed under vacuum. N-(4-(dimethylamino)butyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide was obtained by recrystallization.

[0192] Step 2: Preparation of N-(4-(dimethylamino)butyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide

[0193] Using the N-(4-(dimethylamino)butyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide obtained in the above step 1, a method and procedure similar to that used in step 7 of the above example 1 were performed to obtain the final product, N-(4-(dimethylamino)butyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide (61 mg, 90% yield).

[0194] 1 H NMR (500 MHz, dmso) δ 10.16 (s, 1H), 8.67 (d, J = 4.6 Hz, 1H), 7.82 - 7.78 (m, 1H), 7.50 - 7.44 (m, 1H), 7.21 (s, 2H), 7.16 (s, 1H), 3.65 - 3.51 (m, 5H), 3.27 (d, J = 5.7 Hz, 3H), 3.23 - 3.14 (m, 2H), 3.12 - 3.06 (m, 2H), 2.76 (s, 5H), 1.72 - 1.62 (m, 2H), 1.58 - 1.50 (m, 2H); 13C NMR (126 MHz, dmso) δ 166.95, 161.33, 154.22, 146.92, 139.71, 135.33, 129.71, 129.54, 125.01, 124.13, 117.12, 112.94, 66.62, 56.73, 49.03, 42.59, 38.80, 26.60, 21.77; LC-MS [C 22 H 29 N3O4S] calcd for [M+H], 432.2 found 432.2

[0195] Examples 7 to 21 below were synthesized based on the following reaction scheme 2, and each substituent is specifically described in the reaction scheme. The reagents and conditions used in the reaction scheme 2 are as follows. (h) Methyl 4-bromo-2-methoxybenzoate, (5-formylthiophen-2-yl)boronic acid, 2 M Na2CO3, Pd(PPh3)4, 1,2-DME, 80 ℃, overnight; (i) NaH2PO4H2O, H2O2(30%), NaClO2(80%), Acetonitrile:H2O=3:2, rt, 3 hrs; (j) N,N-Dimethyl diamine, EDC·HCl, HOBt, DIPEA, DMF, rt, overnight; (k) 1 M LiOH, THF, 50 o C, overnight; (l) Amines, HATU, DIPEA, DMF, rt, 3hrs.

[0196] [Reaction Formula 2]

[0197]

[0198] Example 7. Preparation of tert-butyl 4-(4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoyl)piperazine-1-carboxylate (15e)

[0199]

[0200] Step 1: Preparation of methyl 4-(5-formylthiophen-2-yl)-2-methoxybenzoate

[0201] Methyl 4-bromo-2-methoxybenzoate and (5-formylthiophen-2-yl)boronic acid were dissolved in 1,2-dimethoxyethane, and sodium carbonate was added. The mixture was purged with nitrogen to remove oxygen. Tetrakis(triphenylphosphine)-palladium was then added to the mixture, covered with foil, and stirred at 80°C overnight. The metal catalyst was removed through a celite filter. The solution was concentrated under reduced pressure, and the resulting residue was diluted with ethyl acetate and washed with water and brine. The concentrate was purified by silica gel column chromatography to obtain methyl 4-(5-formylthiophen-2-yl)-2-methoxybenzoate (885 mg, 38% yield).

[0202] Step 2: Preparation of 5-(3-methoxy-4-(methoxycarbonyl)phenyl)thiophene-2-carboxylic acid

[0203] Methyl 4-(5-formylthiophen-2-yl)-2-methoxybenzoate was dissolved in a solution of acetonitrile and water in a 3:2 ratio. Sodium phosphate monobasic monohydrate and hydrogen peroxide were added, and 80% sodium chlorite dissolved in water was added at 0°C. The mixture was stirred at room temperature for 3 hours. The mixture was acidified with a 10% hydrochloric acid solution (pH 2–3). The solution was concentrated under reduced pressure, and the resulting residue was diluted with ethyl acetate and washed with water and brine. The above concentrate was purified by silica gel column chromatography without purification, and 5-(3-methoxy-4-(methoxycarbonyl)phenyl)thiophene-2-carboxylic acid was obtained (927 mg, 99% yield).

[0204] Step 3: Preparation of methyl 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoate

[0205] N,N-dimethylpropane-1,3-diamine, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, hydroxybenzotriazole, and N,N-diisopropylethylamine were added to dimethylformamide in which 5-(3-methoxy-4-(methoxycarbonyl)phenyl)thiophene-2-carboxylic acid was dissolved. The mixture was stirred at room temperature overnight. After dimethylformamide was concentrated under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain methyl 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoate (820 mg, 70% yield).

[0206] Step 4: Preparation of 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid

[0207] 1 M lithium hydroxide was added dropwise to a tetrahydrofuran solution containing methyl 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoate at 0°C. The mixture was stirred overnight at 50°C. The mixture was acidified with a 10% hydrochloric acid solution (pH 2–3). After concentration under reduced pressure, the concentrate was purified by silica gel column chromatography. 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid was obtained. (620 mg, 78% yield)

[0208] Step 5: Preparation of tert-butyl 4-(4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoyl)piperazine-1-carboxylate

[0209] 1-Boc-piperazine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine were added to the above 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid. The mixture was stirred at room temperature for three hours. After concentrating dimethylformamide under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, tert-butyl 4-(4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoyl)piperazine-1-carboxylate (66 mg, 22.2% yield).

[0210] 1 H NMR (500 MHz, cd3od) δ 7.70 (d, J = 3.9 Hz, 1H), 7.53 (d, J = 3.9 Hz, 1H), 7.37 (s, 1H), 7.35 (d, J = 4.5 Hz, 1H), 7.30 (d, J = 7.8 Hz, 1H), 3.94 (s, 3H), 3.80 (d, J = 13.8 Hz, 1H), 3.68 (d, J = 10.0 Hz, 1H), 3.53 (d, J = 18.1 Hz, 2H), 3.46 (t, J = 6.7 Hz, 3H), 3.42 (s, 2H), 3.26 (d, J = 5.6 Hz, 1H), 3.02 - 2.94 (m, 2H), 2.73 (s, 6H), 2.03 - 1.94 (m, 23H), 1.47 (s, 9H). 13C NMR (126 MHz, cd3od) δ 168.20, 163.09, 155.90, 154.76, 147.86, 138.01, 136.32, 129.41, 128.54, 124.76, 124.59, 118.36, 108.23, 80.26, 55.73, 54.95, 46.59, 42.67, 41.42, 36.56, 27.17, 25.44; Purity > 91% (RP-HPLC, t R = 9.465 min); LC-MS [C 27 H 38 N4O5S] calcd for [M+H], 531.1 found 531.1.

[0211] Example 8. Preparation of N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(piperazine-1-carbonyl)phenyl)thiophene-2-carboxamide (15a)

[0212]

[0213] The tert-butyl 4-(4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoyl)piperazine-1-carboxylate obtained in Example 7 was dissolved in methanol, and 2 M hydrochloric acid dissolved in diethyl ether was added. The mixture was stirred at 35°C overnight. After concentrating methanol under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(piperazine-1-carbonyl)phenyl)thiophene-2-carboxamide (77 mg, 68.7% yield).

[0214] 1H NMR (500 MHz, dmso) δ 8.60 (s, 1H), 7.74 (d,J= 2.0 Hz, 1H), 7.63 (d,J= 2.1 Hz, 1H), 7.35 - 7.28 (m, 2H), 7.22 (d,J= 7.3 Hz, 1H), 3.88 (s, 3H), 3.53 (dd,J= 40.5, 10.9 Hz, 2H), 3.26 (d,J= 6.0 Hz, 2H), 3.06 (s, 2H), 2.76 - 2.52 (m, 5H), 2.26 (t,J= 6.3 Hz, 2H), 2.14 (s, 6H), 1.95 (d,J= 17.4 Hz, 1H), 1.70 - 1.61 (m, 2H); 13 C NMR (126 MHz, dmso) δ 166.27, 161.22, 155.84, 146.85, 140.01, 135.46, 129.19, 128.98, 126.26, 125.52, 118.55, 108.82, 57.23, 56.14, 48.13, 46.28, 45.84, 45.59, 42.75, 38.07, 27.54; Purity > 99% (RP-HPLC, t R = 1.628 min); LC-MS [C 22 H 30 N4O3S] calcd for [M+H], 431.2 found 431.2

[0215] Example 9. Preparation of N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(4-methylpiperazine-1-carbonyl)phenyl)thiophene-2-carboxamide (15b)

[0216]

[0217] Methylpiperazine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine were added to 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7. The mixture was stirred at room temperature for three hours. After concentrating dimethylformamide under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(4-methylpiperazine-1-carbonyl)phenyl)thiophene-2-carboxamide (10 mg, 3% yield).

[0218] 1 H NMR (500 MHz, dmso) δ 8.59 (t,J= 5.3 Hz, 1H), 7.74 (d,J= 3.8 Hz, 1H), 7.63 (d,J= 3.7 Hz, 1H), 7.34 - 7.28 (m, 2H), 7.22 (d,J= 7.6 Hz, 1H), 3.88 (s, 3H), 3.60 (dd,J= 31.5, 5.3 Hz, 2H), 3.25 (dd,J= 12.8, 6.4 Hz, 2H), 3.13 (d,J= 4.2 Hz, 2H), 2.33 (d,J= 12.7 Hz, 2H), 2.25 (t,J= 6.9 Hz, 3H), 2.18 (s, 4H), 2.13 (s, 5H), 1.68 - 1.60 (m, 2H); 13C NMR (126 MHz, dmso) δ 166.26, 161.17, 155.88, 146.79, 140.12, 135.58, 129.17, 129.01, 126.06, 125.56, 118.52, 108.88, 72.93, 63.50, 57.26, 56.16, 55.16, 46.11, 45.64, 41.37, 38.06, 27.60; Purity > 99% (RP-HPLC, t R = 1.605 min); LC-MS [C 23 H 32 N4O3S] calcd for [M+H], 445.2 found 445.2

[0219] Example 10. Preparation of N-(3-(dimethylamino)propyl)-5-(4-(4-ethylpiperazine-1-carbonyl)-3-methoxyphenyl)thiophene-2-carboxamide (15c)

[0220]

[0221] 1-Ethylpiperazine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine were added to 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7. The mixture was stirred at room temperature for three hours. After concentrating dimethylformamide under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(4-(4-ethylpiperazine-1-carbonyl)-3-methoxyphenyl)thiophene-2-carboxamide (133.3 mg, 42.9% yield).

[0222] 1H NMR (500 MHz, dmso) δ 8.59 (t, J = 5.4 Hz, 1H), 7.73 (d, J = 3.8 Hz, 1H), 7.63 (d, J = 3.8 Hz, 1H), 7.35 - 7.27 (m, 2H), 7.22 (d, J = 7.7) Hz, 1H), 3.87 (s, 3H), 3.60 (d, J = 23.9 Hz, 3H), 3.25 (dd, J = 12.8, 6.5 Hz, 2H), 3.13 (s, 2H), 2.43 - 2.28 (m, 5H), 2.25 (t, J = 7.0 Hz, 3H), 2.13 (s, 5H), 1.68 - 1.60 (m, 2H), 0.99 (t, J = 7.1 Hz, 3H); 13C NMR (126 MHz, dmso) δ 166.21, 161.19, 155.89, 146.81, 140.08, 135.57, 129.17, 129.03, 126.06, 125.56, 118.53, 108.86, 57.25, 56.16, 52.96, 52.50, 51.94, 46.75, 45.62, 41.49, 38.06, 27.57, 12.33; Purity > 99% (RP-HPLC, tR = 1.631 min); LC-MS [C 24 H 34 N4O3S] calcd for [M+H], 459.3 found 459.3

[0223] Example 11. Preparation of N-(3-(dimethylamino)propyl)-5-(4-(4-isopropylpiperazine-1-carbonyl)-3-methoxyphenyl)thiophene-2-carboxamide (15d)

[0224]

[0225] 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7 was added 1-isopropylpiperazine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine. The mixture was stirred at room temperature for three hours. After concentrating dimethylformamide under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(4-(4-isopropylpiperazine-1-carbonyl)-3-methoxyphenyl)thiophene-2-carboxamide (237 mg, 79% yield).

[0226] 1 H NMR (500 MHz, dmso) δ 8.60 (t,J= 5.3 Hz, 1H), 7.74 (d,J= 3.7 Hz, 1H), 7.63 (d,J= 3.7 Hz, 1H), 7.35 - 7.28 (m, 2H), 7.22 (d,J= 7.7 Hz, 1H), 3.87 (s, 3H), 3.58 (d,J= 56.2 Hz, 6H), 3.26 (dd,J= 12.4, 6.3 Hz, 2H), 3.13 (d,J= 8.5 Hz, 2H), 2.70 - 2.62 (m, 2H), 2.46 - 2.35 (m, 2H), 2.33 - 2.24 (m, 2H), 2.15 (s, 4H), 1.71 - 1.61 (m, 2H), 0.96 (d,J= 6.4 Hz, 6H); 13C NMR (126 MHz, dmso) δ 166.16, 161.20, 155.89, 146.83, 140.05, 135.54, 129.19, 129.04, 126.12, 125.55, 118.52, 108.86, 57.17, 56.15, 48.74, 48.27, 47.14, 45.51, 41.88, 38.01, 27.48, 18.43; Purity > 95% (RP-HPLC, t R = 2.499 min); LC-MS [C 25 H 36 N4O3S] calcd for [M+H], 473.3 found 473.3

[0227] Example 12. Preparation of N-(3-(dimethylamino)propyl)-5-(4-(ethylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide (15f)

[0228]

[0229] 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7 was added ethaneamine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine. The mixture was stirred at room temperature for three hours. After dimethylformamide was concentrated under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(4-(ethylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide (43.4 mg, 20.2% yield).

[0230] 1 H NMR (500 MHz, cd3od) δ 7.91 (d,J= 8.4 Hz, 1H), 7.73 (d,J= 3.7 Hz, 1H), 7.53 (d,J= 3.7 Hz, 1H), 7.34 (d,J= 6.6 Hz, 2H), 4.03 (s, 3H), 3.48 (t,J= 6.6 Hz, 2H), 3.45 - 3.40 (m, 2H), 3.19 - 3.12 (m, 2H), 2.87 (s, 6H), 2.04 (dt,J= 13.5, 6.7 Hz, 4H), 1.23 (t,J= 7.2 Hz, 3H); 13 C NMR (126 MHz, cd3od) δ 165.93, 163.15, 158.00, 147.55, 138.19, 137.59, 131.47, 129.58, 124.98, 121.73, 117.94, 108.54, 55.35, 55.28, 42.20, 36.19, 34.32, 24.91, 13.61; Purity > 95% (RP-HPLC, t R = 1.748 min); LC-MS [C 20 H 27 N3O3S] calcd for [M+H], 390.2 found 390.2

[0231] Example 13. Preparation of 5-(4-(diethylcarbamoyl)-3-methoxyphenyl)-N-(3-(dimethylamino)propyl)thiophene-2-carboxamide (15 g)

[0232]

[0233] Diethylamine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine were added to 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7. The mixture was stirred at room temperature for three hours. After dimethylformamide was concentrated under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, 5-(4-(diethylcarbamoyl)-3-methoxyphenyl)-N-(3-(dimethylamino)propyl)thiophene-2-carboxamide (54.7 mg, 23.7% yield).

[0234] 1 H NMR (500 MHz, cd3od) δ 7.66 (d,J= 3.9 Hz, 1H), 7.46 (d,J= 3.8 Hz, 1H), 7.30 (d,J= 10.7 Hz, 2H), 7.19 (d,J= 7.6 Hz, 1H), 3.88 (s,J= 7.4 Hz, 3H), 3.52 (d,J= 49.3 Hz, 2H), 3.40 (t,J= 6.6 Hz, 2H), 3.18 (d,J= 6.2 Hz, 2H), 2.86 - 2.81 (m, 2H), 2.60 (s, 6H), 1.91 (dt,J=14.0, 6.9 Hz, 2H), 1.21 (t,J= 7.1 Hz, 3H), 1.04 (t,J= 7.1 Hz, 3H). 13 C NMR (126 MHz, cd3od) δ 169.22, 163.03, 155.81, 147.95, 137.92, 135.79, 129.40, 127.80, 126.10, 124.45, 118.21, 108.31, 55.95, 54.94, 43.19, 42.93, 39.21, 36.83, 25.67, 12.81, 11.69; Purity > 89% (RP-HPLC, tR = 6.274 min); LC-MS [C 22 H 31 N3O3S] calcd for [M+H], 418.2 found 418.2

[0235] Example 14. Preparation of N-(3-(dimethylamino)propyl)-5-(4-(isopropylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide (15 h)

[0236]

[0237] Isopropylamine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine were added to 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7. The mixture was stirred at room temperature for three hours. After concentrating dimethylformamide under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(4-(isopropylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide (107.2 mg, 44.6% yield).

[0238] 1H NMR (500 MHz, dmso) δ 8.59 (t, J = 5.3 Hz, 1H), 7.93 (d, J = 7.6 Hz, 1H), 7.74 (t, J = 6.3 Hz, 2H), 7.67 (d, J = 3.7 Hz, 1H), 7.40 - 7.30 (m, 2H), 4.06 (td, J = 13.4, 6.7 Hz, 1H), 3.97 (s, 3H), 3.26 (dd, J = 12.9, 6.6 Hz, 2H), 2.25 (t, J = 7.0 Hz, 2H), 2.13 (s, 6H), 1.69 - 1.59 (m, 2H), 1.16 (d, J = 6.5 Hz, 6H). 13 C NMR (126 MHz, dmso) δ 164.00, 161.12, 157.74, 146.45, 140.48, 136.90, 131.59, 129.18, 125.97, 123.71, 118.22, 109.35, 57.25, 56.61, 45.64, 41.30, 38.07, 27.59, 22.81; Purity > 99% (RP-HPLC, t R = 1.885 min); LC-MS [C 21 H 29 N3O3S] calcd for [M+H], 404.2 found 404.2

[0239] Example 15. Preparation of N-(3-(dimethylamino)propyl)-5-(4-(isobutylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide (15i)

[0240]

[0241] Isobutylamine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine were added to 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7. The mixture was stirred at room temperature for three hours. After concentrating dimethylformamide under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(4-(isobutylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide (146.9 mg, 81.8% yield).

[0242] 1 H NMR (500 MHz, dmso) δ 8.59 (t, J = 5.2 Hz, 1H), 8.16 (t, J = 5.6 Hz, 1H), 7.77 - 7.72 (m, 2H), 7.68 (d, J = 3.7 Hz, 1H), 7.39 - 7.32 (m, 2H), 3.97 (s, 3H), 3.26 (dd, J = 12.6, 6.4 Hz, 2H), 3.11 (t, J = 6.3 Hz, 2H), 2.25 (t, J = 6.9 Hz, 2H), 2.13 (s, 6H), 1.88 - 1.76 (m, 1H), 1.70 - 1.59 (m, 2H), 0.90 (d, J = 6.6 Hz, 6H); 13 C NMR (126 MHz, dmso) δ 164.87, 161.13, 157.73, 146.46, 140.49, 136.92, 131.65, 129.18, 125.98, 123.71, 118.23, 109.33, 57.26, 56.59, 46.90, 45.64, 38.07, 28.51, 27.59, 20.55; Purity > 99% (RP-HPLC, t R= 1.975 min); LC-MS [C 22 H 31 N3O3S] calcd for [M+H], 418.2 found 418.2

[0243] Example 16. Preparation of N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(piperidine-1-carbonyl)phenyl)thiophene-2-carboxamide (15j)

[0244]

[0245] Piperidine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine were added to 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7. The mixture was stirred at room temperature for three hours. After concentrating dimethylformamide under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(piperidine-1-carbonyl)phenyl)thiophene-2-carboxamide (90.6 mg, 51.4% yield).

[0246] 1H NMR (500 MHz, dmso) δ 8.58 (t, J = 5.3 Hz, 1H), 7.74 (d, J = 3.8 Hz, 1H), 7.63 (d, J = 3.8 Hz, 1H), 7.34 - 7.27 (m, 2H), 7.20 (d, J = 7.7) Hz, 1H), 3.87 (s, 3H), 3.62 (dd, J = 11.9, 6.2 Hz, 1H), 3.52 (dd, J = 11.8, 6.0 Hz, 1H), 3.26 (dd, J = 12.9, 6.6 Hz, 2H), 3.11 (t, J = 4.5 Hz, 2H), 2.25 (t, J = 7.0 Hz, 2H), 2.13 (s, 6H), 1.64 (dt, J = 14.5, 7.2 Hz, 2H), 1.61 - 1.49 (m, 4H), 1.41 (ddd, J = 16.4, 10.7, 5.3 Hz, 2H); 13 C NMR (126 MHz, dmso) δ 166.06, 161.18, 155.85, 146.87, 140.04, 135.34, 129.15, 128.73, 126.63, 125.48, 118.50, 108.82, 57.25, 56.13, 47.66, 45.63, 42.13, 38.06, 27.59, 26.36, 25.76, 24.49; Purity > 96% (RP-HPLC, t R = 2.393 min); LC-MS [C 23 H 31 N3O3S] calcd for [M+H], 430.3 found 430.3

[0247] Example 17. Preparation of N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(pyrrolidine-1-carbonyl)phenyl)thiophene-2-carboxamide (15k)

[0248]

[0249] In step 4 of Example 1, 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid was obtained, and pyrrolidine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for three hours. After dimethylformamide was concentrated under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(pyrrolidine-1-carbonyl)phenyl)thiophene-2-carboxamide. (107 mg, 59.8%)

[0250] 1 H NMR (500 MHz, dmso) δ 8.58 (t, J = 5.2 Hz, 1H), 7.74 (d, J = 3.4 Hz, 1H), 7.63 (d, J = 3.4 Hz, 1H), 7.35 - 7.27 (m, 2H), 7.24 (d, J = 7.7) Hz, 1H), 3.89 (s, 3H), 3.43 (t, J = 6.7 Hz, 2H), 3.26 (dd, J = 12.7, 6.4 Hz, 2H), 3.13 (t, J = 6.4 Hz, 2H), 2.25 (t, J = 6.9 Hz, 2H), 2.13 (s, 6H), 1.85 (dt, J = 13.1, 6.6 Hz, 2H), 1.82 - 1.75 (m, 2H), 1.68 - 1.61 (m, 2H); 13C NMR (126 MHz, dmso) δ 166.16, 161.18, 155.86, 146.86, 140.07, 135.47, 129.16, 128.82, 127.72, 125.51, 118.42, 109.05, 57.26, 56.17, 47.46, 45.63, 38.06, 27.60, 25.85, 24.49; Purity > 99% (RP-HPLC, t R = 2.143 min); LC-MS [C 22 H 29 N3O3S] calcd for [M+H], 416.2 found 416.2

[0251] Example 18. Preparation of 5-(4-(benzylcarbamoyl)-3-methoxyphenyl)-N-(3-(dimethylamino)propyl)thiophene-2-carboxamide (15 l)

[0252]

[0253] Benzylamine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine were added to 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7. The mixture was stirred at room temperature for three hours. After dimethylformamide was concentrated under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, 5-(4-(benzylcarbamoyl)-3-methoxyphenyl)-N-(3-(dimethylamino)propyl)thiophene-2-carboxamide (220 mg, 84.1% yield).

[0254] 1H NMR (500 MHz, dmso) δ 8.73 (t, J = 6.0 Hz, 1H), 8.60 (t, J = 5.3 Hz, 1H), 7.80 (d, J = 8.0 Hz, 1H), 7.75 (d, J = 3.8 Hz, 1H), 7.69 (d, J = 3.8 Hz, 1H), 7.39 (s, 1H), 7.35 (dd, J = 13.5, 6.5 Hz, 5H), 7.24 (dq, J = 8.5, 4.1 Hz, 1H), 4.51 (d, J = 6.0 Hz, 2H), 3.99 (s, 3H), 3.26 (dd, J = 13.0, 6.5 Hz, 2H), 2.25 (t, J = 7.0 Hz, 2H), 2.13 (s, 6H), 1.69 - 1.61 (m, 2H); 13 C NMR (126 MHz, dmso) δ 164.98, 161.12, 157.93, 146.40, 140.57, 140.11, 137.20, 131.86, 129.20, 128.70, 127.45, 127.08, 126.08, 123.13, 118.23, 109.34, 57.25, 56.57, 45.64, 43.02, 38.07, 27.59; Purity > 99% (RP-HPLC, t R = 2.056 min); LC-MS [C 25 H 29 N3O3S] calcd for [M+H], 452.2 found 452.2

[0255] Example 19. Preparation of N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(phenethylcarbamoyl)phenyl)thiophene-2-carboxamide (15m)

[0256]

[0257] 2-Phenylethanamine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine were added to 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7. The mixture was stirred at room temperature for three hours. After concentrating dimethylformamide under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(phenethylcarbamoyl)phenyl)thiophene-2-carboxamide (186 mg, 69% yield).

[0258] 1 H NMR (500 MHz, dmso) δ 8.58 (t, J = 4.8 Hz, 1H), 8.18 (t, J = 4.2 Hz, 1H), 7.79 (d, J = 8.1 Hz, 1H), 7.73 (d, J = 3.7 Hz, 1H), 7.66 (d, J = 3.6 Hz, 1H), 7.36 - 7.28 (m, 3H), 7.26 (d, J = 7.1 Hz, 1H), 7.21 (t, J = 7.0 Hz, 1H), 3.89 (s, 2H), 3.51 (dd, J = 12.3, 6.1 Hz, 2H), 3.24 (dd, J = 11.1, 4.9 Hz, 2H), 2.82 (t, J = 6.9 Hz, 2H), 2.23 (t, J = 6.8 Hz, 2H), 2.11 (s, 4H), 1.63 (dt, J = 13.6, 6.7 Hz, 2H); 13C NMR (126 MHz, dmso) δ 164.54, 161.12, 157.87, 146.35, 140.58, 139.90, 137.21, 131.94, 129.19, 129.15, 128.84, 126.61, 126.09, 122.74, 118.26, 109.34, 57.24, 56.48, 45.62, 41.15, 38.07, 35.44, 27.57; Purity > 99% (RP-HPLC, t R = 2.144 min); LC-MS [C 26 H 31 N3O3S] calcd for [M+H], 466.3 found 466.3

[0259] Example 20. Preparation of N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(pyridin-3-ylcarbamoyl)phenyl)thiophene-2-carboxamide (15n)

[0260]

[0261] 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7 was added 3-aminopyridine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine. The mixture was stirred at room temperature for three hours. After concentrating dimethylformamide under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(pyridin-3-ylcarbamoyl)phenyl)thiophene-2-carboxamide (99.5 mg, 41.2% yield).

[0262] 1 H NMR (500 MHz, dmso) δ 10.32 (s, 1H), 8.88 (s, 1H), 8.61 (t, J = 5.0 Hz, 1H), 8.31 (d, J = 4.0 Hz, 1H), 8.19 (d, J = 8.0 Hz, 1H), 7.77 (d, J = 3.5 Hz, 1H), 7.72 (d, J = 4.3 Hz, 2H), 7.45 (s, 1H), 7.39 (t, J = 7.4 Hz, 2H), 4.01 (s, 3H), 3.27 (dd, J = 12.7, 6.5 Hz, 2H), 2.25 (t, J = 6.9 Hz, 2H), 2.14 (s, 6H), 1.70 - 1.61 (m, 2H); 13C NMR (126 MHz, dmso) δ 164.80, 161.12, 157.59, 146.32, 144.96, 141.88, 140.71, 137.44, 136.04, 131.26, 129.21, 127.13, 126.21, 124.25, 124.06, 118.29, 109.36, 57.25, 56.66, 45.64, 38.08, 27.59; Purity > 99% (RP-HPLC, t R = 1.718 min); LC-MS [C 23 H 26 N4O3S] calcd for [M+H], 439.2 found 439.2

[0263] Example 21. Preparation of N-(3-(dimethylamino)propyl)-5-(4-((furan-2-ylmethyl)carbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide (15o)

[0264]

[0265] 4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in step 4 of Example 7 was added with furfurylamine, hexafluorophosphate azabenzotriazole tetramethyl uronium, and N,N-diisopropylethylamine. The mixture was stirred at room temperature for three hours. After concentrating dimethylformamide under reduced pressure, the concentrate was purified by silica gel column chromatography to obtain the final product, N-(3-(dimethylamino)propyl)-5-(4-((furan-2-ylmethyl)carbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide (140 mg, 60.9% yield).

[0266] 1 H NMR (500 MHz, dmso) δ 8.61 (dd, J = 11.5, 5.4 Hz, 2H), 7.81 (d, J = 8.0 Hz, 1H), 7.75 (d, J = 3.7 Hz, 1H), 7.69 (d, J = 3.7 Hz, 1H), 7.58 (s, 1H), 7.40 - 7.33 (m, 2H), 6.40 (s, 1H), 6.27 (s, 1H), 4.49 (d, J = 5.6 Hz, 2H), 3.98 (s, 3H), 3.26 (dd, J = 12.5, 6.3 Hz, 2H), 2.24 (t, J = 6.9 Hz, 2H), 2.13 (s, 6H), 1.68 - 1.61 (m, 2H). 13C NMR (126 MHz, dmso) δ 164.76, 161.12, 157.95, 152.90, 146.33, 142.40, 140.62, 137.35, 131.95, 129.19, 126.13, 122.67, 118.24, 110.93, 109.35, 107.02, 57.26, 56.60, 45.64, 38.08, 36.60, 27.59; Purity > 99% (RP-HPLC, t R = 1.874 min); LC-MS [C 23 H 27 N3O4S] calcd for [M+H], 442.2 found 442.2

[0267] Example 22. Preparation of N-(3-(dimethylamino)propyl)-5-(4-hydroxy-3-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide (29)

[0268]

[0269] Example 22 was synthesized based on the following reaction scheme 3, and the reagents and conditions used in reaction scheme 3 are as follows. (a) Di-tert-butyldicarbonate, 4-DMAP, DIPEA, DCM, 55°C, overnight; (b) Boronic acid, Pd(PPh3)4, Cs2CO3, DME / Water (1:1), 68°C, overnight; (c) NaOH aqueous solution, MeOH, THF (1:1:3), 50°C, 30 min; (d) Amines, EDC·HCl, HOBt, TEA, DMF, rt, overnight; (e) TFA, DCM, rt, 2.5 hrs; (f) Aliphatic amine, EDC·HCl, HOBt, TEA, DMF, rt, overnight; (g) BBr3, DCM, -78°C→rt, 1.5 hrs.

[0270] [Reaction Formula 3]

[0271]

[0272] Step 1: Preparation of tert-butyl 5-bromothiophene-2-carboxylate

[0273] 5-Bromothiophene-2-carboxylic acid was dissolved in dichloromethane in an oven-dried, sealed tube. 4-(dimethylamino)pyridine and di-tert-butyl dicarbonate were added to this solution at room temperature. N,N-diisopropylethylamine was then added sequentially at the same temperature. The mixture was stirred overnight at 55°C. The mixture was cooled to room temperature, diluted with distilled water, and extracted three times with dichloromethane. The combined organic phases were dried over magnesium sulfate. The solvent was evaporated in vacuo. The residue was purified by flash column chromatography using a hexane-ethyl acetate (19:1) system, yielding a pale yellow oil.

[0274] Step 2: Preparation of tert-butyl 5-(4-methoxy-3-(methoxycarbonyl)phenyl)thiophene-2-carboxylate

[0275] The tert-butyl 5-bromothiophene-2-carboxylate obtained in Step 1 was added to a sealed, oven-dried tube at room temperature. (3-Methoxy-4-(methoxycarbonyl)phenyl)boronic acid and cesium carbonate were added at room temperature. Dimethyl ether and water were also added. The mixture was bubbled with an inert gas to remove dissolved oxygen. Tetrakis(triphenylphosphine)palladium was then added at room temperature. The mixture was heated to 68°C and stirred overnight. The reaction mixture was cooled to room temperature, diluted with distilled water, and extracted with ethyl acetate. The combined organic phases were dried over magnesium sulfate. The solvent was removed under vacuum. The residue was purified by flash column chromatography using a hexane-ethyl acetate (14:1) system to obtain a brown oil.

[0276] Step 3: Preparation of 5-(5-(tert-butoxycarbonyl)thiophen-2-yl)-2-methoxybenzoic acid

[0277] tert-Butyl 5-(4-methoxy-3-(methoxycarbonyl)phenyl)thiophene-2-carboxylate obtained in Step 2 was added to a mixture of tetrahydrofuran and methanol with 1 molar aqueous sodium hydroxide solution. The mixture was stirred at 50°C for 30 minutes. After confirming the reaction, the mixture was diluted with water and acidified with a 10% aqueous hydrogen chloride solution. It was then extracted three times with ethyl acetate. The combined organic phases were dried over magnesium sulfate. The solvent was removed under vacuum to give a white solid, which was used in the next reaction without further purification.

[0278] Step 4: Preparation of tert-butyl 5-(4-methoxy-3-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxylate

[0279] 5-(5-(tert-butoxycarbonyl)thiophen-2-yl)-2-methoxybenzoic acid obtained in the above step 3 was dissolved in N,N-dimethylformamide, and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, hydroxybenzotriazole, and triethylamine were added at room temperature. The mixture was stirred at room temperature. After the reaction was completed, the reaction solution was diluted with dilute water and extracted three times with ethyl acetate. The organic solvent was washed with water and brine to remove N,N-dimethylformamide, and dried over magnesium sulfate. The solvent was removed in vacuo to give a brown oil, which was used in the next reaction without further purification.

[0280] Step 5: Preparation of 5-(4-methoxy-3-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxylic acid

[0281] The tert-butyl 5-(4-methoxy-3-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxylate obtained in Step 4 was dissolved in dichloromethane solvent at 0°C, and trifluoroacetic acid was added. The mixture was stirred at room temperature for 2.5 hours. After monitoring the disappearance of the reaction mass, the mixture was diluted with dilute water and acidified with a 10% aqueous hydrogen chloride solution. It was then extracted three times with dichloromethane. The organic solvent was dried over magnesium sulfate. The solvent was removed under vacuum to obtain a white solid, which was used in the next reaction without further purification.

[0282] Step 6: Preparation of N-(3-(dimethylamino)propyl)-5-(4-methoxy-3-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide

[0283] 5-(4-methoxy-3-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxylic acid obtained in Step 5 above was dissolved in N,N-dimethylformamide, and then N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, hydroxybenzotriazole, N,N-dimethyl-1,3-propanediamine, and triethylamine were added at room temperature. The mixture was stirred at room temperature overnight. After the reaction product appeared, it was diluted with distilled water and extracted. The organic solvent was washed with water and brine, and the organic phase was dried over magnesium sulfate to remove DMF. The solvent was removed under vacuum. A brown solid was obtained by recrystallization.

[0284] Step 7: Preparation of N-(3-(dimethylamino)propyl)-5-(4-hydroxy-3-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide

[0285] N-(3-(dimethylamino)propyl)-5-(4-methoxy-3-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide obtained in step 6 above was dissolved in dimethyl chloro, and then 1 molar tribromoboron dichloro solution was slowly added at -78°C. The mixture was stirred at -78°C for 1.5 hours. After the product appeared, it was left at room temperature. The mixture was diluted with dichloromethane, and a brown solid was obtained by filtration.

[0286] 1H NMR (500 MHz, cd3od) δ 7.65 (d, J = 4.0 Hz, 1H), 7.41 (d, J = 3.5 Hz, 1H), 7.28 - 7.23 (m, 2H), 7.26 (s, 1H), 3.77 - 3.63 (m, 6H), 3.48 (t, J = 6.4 Hz, 3H), 3.34 (d, J = 10.9 Hz, 2H), 3.26 - 3.21 (m, 2H), 2.93 (s, 6H), 2.10 - 2.02 (m, 2H); LC-MS [C 21 H 27 N3O4S] calcd for [M+H], 418.2 found 418.2.

[0287] <Experimental Example>

[0288] Experimental Example 1. Experimental Preparation and Experimental Method

[0289] 1) Cell culture

[0290] Human brain microvascular endothelial cells (HBMVECs) were purchased from Cell Systems. Plates were coated with adhesion factor (1:1,000 in PBS) and cultured in cell culture medium (M-199 medium containing 20% ​​fetal bovine serum (FBS), 5 U / ml heparin, 3 ng / ml recombinant human fibroblast growth factor-basic, 100 U / ml penicillin, and 100 μg / ml streptomycin) at 37°C in a 5% CO2 incubator. Human leukemia-60 cells were purchased from the Korean Cell Line Bank and cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, penicillin, and streptomycin.

[0291] 2) Measurement of KDM4 enzyme activity

[0292] Enzyme activity was performed according to the manufacturer's instructions (Abcam, ab113462), and recombinant KDM4C protein among KDM4 enzymes was used. Recombinant KDM4C protein, substrate, experimental buffer (prepared by mixing experimental buffer and co-factors 1, 2, and 3 in a ratio of 100:1:1:1), and compound to be measured were mixed, placed in each well of a microplate container coated with trimethyl histone H3K9 substrate, and incubated in a 37℃ incubator for 90 minutes, followed by washing three times with 1x wash buffer (10x wash buffer diluted with water). 50 μl of capture antibody diluted 1:1,000 in 1x wash buffer was added, and incubated at room temperature for 1 hour. After washing three times with 1x wash buffer, 50 μl of detection antibody diluted 1:2,000 with 1x wash buffer was added to each well and incubated at room temperature for 1 hour. After washing four times with 1x wash buffer, 50 μl of fluorescence development solution (fluoro developer and fluoro enhancer diluted 1:1:500 with fluoro diluter) was added to each well and incubated at room temperature for 2 to 4 minutes. Care should be taken to avoid exposure to light after adding fluorescence development solution. Enzyme activity was measured at 530 / 590 mm (absorption / emission) in a fluorescence microplate reader.

[0293] 3) Protein quantification

[0294] Protein concentration was performed in duplicate in 96-well plates according to the manufacturer's method using the BCA quantitation kit (Pierce, 23235). After protein extraction from cells, 2 μl of the obtained sample was aliquoted into each well, filled to 20 μl with DW, and a solution of Reagent A and B (50:1) was added to each well (180 μl) to make a total volume of 200 μl. The plate was then wrapped in foil to block light and incubated in a 37°C incubator for 30 minutes. The absorbance was measured at 562 nm using a microplate reader.

[0295] 4) Western blot analysis

[0296] HBMVECs in the culture dish were washed with cold PBS, and the cells were lysed with RIPA buffer (20 mM Tris-HCl, pH 7.4 containing 150 mM NaCl, 1 mM ethylenediaminetetraacetic acid (EDTA), 1 mM ethylene glycol tetraacetic acid (EGTA), 1% Triton X-100, and protein phosphatase inhibitor cocktail (Roche)). The cells were transferred to a 1.5 ml tube using a cell scraper and incubated at 4°C for 30 minutes.

[0297] Total protein (10 μg) was mixed with 4x LDS sample buffer, RIPA buffer, and 20x dithiothreitol (DTT), heated in a 100°C block for 5 minutes, and then reacted at 4°C for 5 minutes. The sample was loaded into a pre-made 10% SDS-polyacrylamide gel and electrophoresed. The protein in the gel was then transferred to a polyvinylidene difluoride membrane. The membrane was incubated with a blocking solution (4% skim milk in TBS-T) for 1 hour at room temperature, and then reacted with primary antibodies [VCAM1 (1:1,000, Abcam), α-tubulin (1:5,000, Millipore), 6E10 (1:1,000, Biolegend)] appropriately diluted in blocking solution at 4°C overnight. The membrane was washed with TBS-T and reacted with a secondary antibody [anti-rabbit or anti-mouse horseradish peroxidase-conjugated IgG (1:5,000)] diluted in blocking solution at room temperature for 1 hour. The membrane was then washed with TBS-T and reacted with ECL solution to develop color. Bands were identified using LAS equipment.

[0298] 5) Cell adhesion quantification method

[0299] HBMVEC cultured in a 24-well plate (Example 1, II-01) were pretreated for 1 hour, cultured in a 37°C cell incubator, treated with TNFα (10 ng / ml), and cultured in a cell incubator for 8 hours. Meanwhile, HL-60 cells (in RPMI 1640 medium) fluorescently labeled by reacting with 5 μM CellTracker Green CMFDA (Thermo) for 10 minutes were dispensed into each well of HBMVEC cells treated with TNFα (500 μl) and cultured in a 37°C incubator for 1 hour. The cells were washed three times with warm PBS, and images were taken using a fluorescence microscope to quantify the attached cells.

[0300] 6) Genetically modified animals and genotyping

[0301] 5xFAD mice are genetically modified animals that carry three human APP gene mutations found in patients with hereditary Alzheimer's disease (Sweden; K670N / M671L, Florida; I716V, London; V717I) and mutations in human presinilin1 (PS1) (M146L, L286V). 5xFAD mice were purchased from Jackson Laboratory (Sacramento, CA, USA) and maintained by crossing hemizygous transgenic mice with B6SJL F1 mice. The genotypes of transgenic mice were identified using polymerase chain reaction. Non-transgenic siblings were used as negative controls.

[0302] For genotyping, genomic DNA was extracted from a portion of the tail of 1-month-old mice and EzDirect was used according to the protocol provided by the company. TMThe presence of the human PS1 gene was confirmed using the Mouse Direct PCR Kit (Wizbiosolutions, Inc.). Animals showing a band by polymerase chain reaction (PCR) analysis using the following primers were considered positive for the Tg genotype: human PS1 forward, 5'-AAT AGA GAA CGG CAG GAG CA-3' (SEQ ID NO: 1); and human PS1 reverse, 5'-GCC ATG AGG GCA CTA ATC AT-3' (SEQ ID NO: 2).

[0303] The 5xFAD mice used in this experiment exhibit extracellular amyloid deposition and gliosis beginning at approximately 2 months of age, spatial working memory impairment beginning at approximately 4-5 months of age, and neuronal loss in several brain regions beginning at approximately 6 months of age. All experiments were performed using 7-month-old 5xFAD and WT mice. Mice were housed in cages of one or two with a 12-h light / dark cycle. The treatment and maintenance of experimental animals were performed in accordance with the approved guidelines for animal care and use (ID: DKU-22-078).

[0304] 7) Y-maze test

[0305] The Y-maze test is a method for measuring the willingness of rats to explore a novel environment. Generally, rats prefer to explore new arms / areas rather than returning to previously visited areas within the maze. The Y-maze consists of a central arm and two lateral arms. Each rat was placed at the end of one arm and allowed to move freely around the maze during an 8-minute session. The rat's movements, i.e., a series of entries, were recorded by a camera. An entry was considered to be a series of entries when the rat entered more than halfway into a particular arm. In the analysis, cognitive function was determined to be positive if the rat entered three different arms consecutively in triplicate.

[0306] 8) Manual subcutaneous examination

[0307] The passive subcutaneous test is a fear-training experiment used to study short-term and long-term memory in an associative manner. This experiment exploits the rat's tendency to avoid bright environments and move toward dark ones. The experiment begins with a shuttle box apparatus, divided into two compartments by an extractable door. One compartment is brightly lit, while the other has dark, opaque walls and a roof, and a metal grid on the floor that delivers a weak 0.3 mA electric shock. On the first day, the rat is placed in the bright compartment. After 30 seconds, the partition blocking the dark compartment is opened, allowing the rat to enter. Upon entering, the partition is closed again, delivering a 1-second electric shock (0.3 mA). After a 1-minute wait, the rat is returned to its home cage. The next day, the rat is placed in the bright compartment in the same manner. After 30 seconds, the partition is removed. The time it takes the rat to enter the dark compartment is measured, up to a maximum of 600 seconds.

[0308] 9) Open field test

[0309] In the open field test, mice were placed in the center of a dark box (40 × 40 × 40 cm) and allowed to move freely. The movement parameters of the mice were recorded and analyzed by a video camera connected to Noldus' EthoVision XT video tracking software. The total distance moved, the time spent in the open field, and the time taken to arrive at the open field (the time taken by the mouse to reach the location on the first attempt) were analyzed.

[0310] 10) Brain tissue preparation

[0311] After completing the behavioral experiment, rats were anesthetized and perfused with 10 mM PBS (pH 7.4). The brains were then separated, weighed, and cut into left and right halves for storage. For immunohistochemical analysis, the right hemisphere was fixed in 4% paraformaldehyde prepared in PBS for 1 h and then preserved in a 30% sucrose solution prepared in PBS until equilibrium was reached. The brains were frozen in Tissue-Tek OCT compound (Sakura Finetek) and sectioned into 20 μm-thick coronal sections using a Cryo-cut device (Leica). The sections were stored in storage buffer (10 mM PBS containing 30% glycerol and 30% ethylene glycol, pH-7.2) at -80°C.

[0312] 11) DAB staining

[0313] Brain sections were washed twice with PBS and treated with 0.3% H2O2 solution for 30 min. These sections were washed again with PBS and treated with blocking solution (PBS containing 1% BSA and 0.1% TritonX-100) for 1 h at room temperature. The sections were treated with primary antibodies [6E10 (1:500, Biolegend), GFAP (1:250; DAKO), Iba-1 (1:500; WAKO), NeuN (1:100; Merck)] for 18 h at 4°C. After washing, the sections were treated with biotin-conjugated secondary antibodies (1:250, biotin antibody: PBS-T) and incubated for 3 h at room temperature. After washing again, the sections were treated with Vectastain ABC solution (Vector laboratories; solution A: solution B:PBS = 1:1:500), then treated with DAB solution (Sigma-Aldrich, solution 1: solution 2 = 1:200) and stained by light. The stained sections were mounted on slides, mounted with mounting solution, and covered with a coverslip. Digital images were taken using an inverted microscope (Olympus) and captured at 200x magnification using ImageJ software (NIH).

[0314] 12) Quantitative and statistical analysis

[0315] All results are expressed as mean ± SEM. Data were analyzed using one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test. Significant differences are indicated by *, p < 0.05; **, p < 0.01, or ***, p < 0.001. Significant differences between the two groups were analyzed using Student's t-test. All statistical analyses were performed using GraphPad Prism 7 software (GraphPad Software, Inc.).

[0316] Experimental Example 2. Evaluation of KDM4C enzyme activity

[0317] The inhibitory effect of the compounds according to Examples 1 to 22 on KDM4 activity was investigated using a KDM4 enzyme activity assay kit (Abcam, ab113462), and the results are shown in Figure 1 and Table 1. The control group is a comparative group that did not use the compound.

[0318] ExampleActivity (%)P. value(vs. control)ExampleActivity (%)P. value(vs. control)Control100.0 ± 6.8-Example 10(II-15)52.6 ± 3.50.003Example 1(II-01)57.1 ± 0.4<0.001Example 8(II-16)56.9 ± 14.20.018Example 2(II-02)73.5 ± 11.60.027Example 18(II-17)52.4 ± 6.20.004Example 3(II-03)66.4 ± 4.40.002Example 19(II-18)53.3 ± 5.30.004Example 7(II-09)68.3 ± 27.30.13Example 14(II-19)48.6 ± 6.6 0.004 Example 12 (II-10) 64.1 ± 21.4 0.062 Example 20 (II-20) 52.6 ± 7.3 0.005 Example 9 (II-11) 70.5 ± 25.1 0.129 Example 15 (II-21) 40.1 ± 1.1 0.001 Example 13 (II-12) 69.5 ± 3.3 0.011 Example 21 (II-22) 37.1 ± 9.5 0.003 Example 16 (II-13) 59.5 ± 2.3 0.005 Example 17 (II-23) 41.4 ± 4.6 0.002 Example 11 (II-14) 63.9 ± 4.2 0.007

[0319] According to Fig. 1 and Table 1, the ID50 of Example 1 (II-01) was measured to be approximately 1 μM, and the degree of enzyme inhibition at a single concentration of 1 μM was also measured for other compounds. It was confirmed that Example 1 (II-01) reduced KDM4C activity by approximately 47%. It was confirmed that most of the remaining compounds effectively inhibited the DM4 enzyme (21% to 63%).

[0320] Experimental Example 3. Evaluation of VCAM1 expression inhibition

[0321] The effect of the compound according to Example 1 on the expression of VCAM1 and ICAM1 in HBMVECs was confirmed by Western blotting, and the results are shown in Fig. 2.

[0322] According to Fig. 2, TNFα increases the expression of ICAM1 and VCAM1 in various endothelial cells, and in this experiment, the expression of ICAM1 and VCAM1 was increased when TNFα (10 ng / ml, 8 hours) was treated. Treatment with the compound (II-01) according to Example 1 inhibited the increase in VCAM1 expression induced by TNFα, but did not inhibit the expression of ICAM1. These results indicate that the compound according to the present invention selectively inhibits only VCAM1.

[0323] Experimental Example 4. Cell Adhesion Evaluation

[0324] Leukocytes cross the blood-brain barrier and migrate into brain tissue under various environmental conditions, aided by vascular adhesion molecules and chemokines. Therefore, we investigated whether the increase in VCAM1 induced by TNF-α treatment alters leukocyte adhesion to vascular cells and investigated the effects of compounds on adhesion. The results are presented in Figure 3.

[0325] According to Fig. 3, the adhesion of HL-60 cells, a type of white blood cell, to HBMVECs was investigated, and TNF-α (10 ng / ml, 8 hours) increased the adhesion of fluorescently labeled HL-60 cells to HBMVECs, and this increase was dose-dependently reduced by the compound (II-01) according to Example 1. Therefore, it was confirmed that the inhibitory effect of the compound (II-01) according to Example 1 on the increase in VCAM1 expression induced by TNF-α was linked to the function of HBMVECs cells. In other words, blocking the adhesion of white blood cells to endothelial cells by inhibiting VCAM1 expression is expected to suppress the aggravation of this pathological mechanism.

[0326] Experimental Example 5. Evaluation of Cognitive Function Improvement

[0327] In order to examine the effect of the compound (II-01) according to Example 1 on cognitive function, various behavioral experiments were performed using the dementia model animal 5xFAD, and the results are shown in Figures 4a to 4e.

[0328] According to Figs. 4a to 4e, 6-month-old 5xFAD and WT mice were administered intravenously with a solvent (saline) and the compound (II-01) according to Example 1 (0.345 mg / kg (Low; L), 1 mg / kg (High; H), dissolved in DMSO) 10 times at 3-day intervals (total 30 days) (Fig. 4a). Food intake and body weight were measured during the administration period to evaluate the toxicity of the compound (II-01) according to Example 1 (Fig. 4b), and there was no significant difference in food intake and body weight between the non-administered and administered groups.

[0329] Additionally, the Y-maze and passive avoidance test (PAT) were used to test the memory of the mice. The open field test, which monitors the locomotor ability of the mice, showed no difference in the total distance traveled between the groups, suggesting no difference in general activity (Fig. 4c). The spontaneous alternation rate in the Y-maze results is known to represent hippocampal-dependent spatial memory. The 5xFAD-solvent group showed a significantly reduced alternation rate compared to the WT-solvent group. However, when the compound (II-01) according to Example 1 was administered to 5xFAD, the reduced alternation rate was restored, and the effect was observed in animals administered not only at high doses but also at low doses (Fig. 4d). PAT evaluates long-term memory by the time it takes for mice to enter a dark room. The 5xFAD-solvent group took less time than the WT-solvent group, but administration of the compound (II-01) according to Example 1 restored this time to the level of the WT-solvent group in both low- and high-dose cases (Fig. 4e). Therefore, it was found that the compound according to the present invention improves memory function in 5xFAD mice, a dementia model animal.

[0330] Experimental Example 6. Evaluation of Amyloid Plaque Deposition

[0331] The effect on amyloid plaque deposition, one of the important pathological phenomena of Alzheimer's disease, was confirmed, and the results are shown in Figures 5a to 5e.

[0332] According to Figures 5a to 5e, in the cerebral cortex and hippocampus, the 5xFAD-solvent group showed an increase in amyloid plaques compared to the WT-solvent group. Meanwhile, when the compound (II-01) according to Example 1 was administered to 5xFAD mice, it was confirmed that amyloid plaques were significantly reduced in both the cerebral cortex and hippocampus compared to the solvent-treated 5xFAD mice. These results showed a similar reducing effect in low-dose administered animals as in high-dose administered animals.

[0333] As a result, the compound of the present invention (i.e., a compound inhibiting KDM4 activity) reduced TNFα-induced VCAM1 expression in HBMVECs and blocked leukocyte adhesion to blood vessels. Furthermore, administration of the compound of the present invention was confirmed to have the effect of reducing brain inflammation, restoring cognitive function, and reducing the amount of amyloid-beta plaques in Alzheimer's disease mice. This suggests that the compound of the present invention has potential as a therapeutic agent for Alzheimer's disease.

[0334] The specification omits detailed descriptions of matters that would be readily apparent and inferred by those skilled in the art. Furthermore, various modifications, other than the specific examples described herein, are possible without altering the technical spirit or essential configuration of the invention. Therefore, the present invention may be practiced in ways other than those specifically described and exemplified herein, as will be readily apparent to those skilled in the art.

Claims

1. A compound represented by the following chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, R1 and R2 may be the same or different, and each independently represent unsubstituted or substituted C 1-5 It is alkyl; n is an integer from 2 to 10; R3 is -NRaRb, NH-Rc, or unsubstituted or substituted C containing at least one N. 2-8 is heterocycloalkyl; Ra and Rb may be the same or different, and each independently represents -H or unsubstituted or substituted C. 1-5 It is alkyl; Rc is unsubstituted or substituted C 1-5 Alkyl, unsubstituted or substituted C 6-12 Aryl, unsubstituted or substituted -(C 1-5 alkyl)-(C 6-12 Aryl), unsubstituted or substituted C 2-12 Heteroaryl, unsubstituted or substituted -(C 1-5 alkyl)-(C 2-12 heteroaryl), unsubstituted or substituted C 2-8 Heterocycloalkyl, or unsubstituted or substituted -(C 1-5 alkyl)-(C 2-8 heterocycloalkyl); R4 is halogen, C 1-3 Alkyl, -O-(C 1-3 alkyl), -CN, -OH, -NH2, -NO2 or C 1-3 It is haloalkyl.

2. In paragraph 1, R3 is any one selected from the group consisting of -NRaRb, NH-Rc, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, imidazolidinyl, thiazolidinyl, tetrahydrofuranyl, azepanyl, diazepanyl, oxazepanyl and thiazepanyl, A compound represented by the formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein R3 is unsubstituted or substituted with at least one halogen, C1-C5 alkyl, C1-C5 haloalkyl, -O-C1-C5 haloalkyl, -O-C1-C5 alkyl, -C(=O)-(C1-C5 alkyl), -C(=O)-O-(C1-C5 alkyl), -OH, -NO2, or -NH2.

3. In paragraph 1, R3 is any one selected from the group consisting of -NRaRb, NH-Rc, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, imidazolidinyl, thiazolidinyl, tetrahydrofuranyl, azepanyl, diazepanyl, oxazepanyl and thiazepanyl, Ra and Rb may be the same or different, and are each independently methyl, ethyl, propyl or isopropyl; Rc is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, phenyl, -(C 1-5 Alkyl)-phenyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, furanyl, isoxazolyl, oxazolyl, oxadiazolyl, thiophenyl, isothiazolyl, thiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, -(C 1-5 alkyl)-pyrrolyl, -(C 1-5 Alkyl)-pyrazolyl, -(C 1-5 alkyl)-imidazolyl, -(C 1-5 Alkyl)-triazolyl, -(C 1-5 alkyl)-furanyl, -(C 1-5 Alkyl)-isoxazolyl, -(C 1-5 alkyl)-oxazolyl, -(C 1-5 Alkyl)-oxadiazolyl, -(C 1-5 alkyl)-thiophenyl, -(C 1-5 Alkyl)-isothiazolyl, -(C 1-5 Alkyl)-thiazolyl, -(C 1-5 Alkyl)-thiadiazolyl, -(C 1-5 alkyl)-pyridinyl, -(C 1-5 alkyl)-pyridazinyl, -(C 1-5 alkyl)-pyrimidinyl, -(C 1-5 alkyl)-pyrazinyl, -(C 1-5 Alkyl)-triazinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, imidazolidinyl, thiazolidinyl, tetrahydrofuranyl, azepanyl, diazepanyl, oxazepanyl, thiazepanyl, -(C 1-5 alkyl)-azetidinyl, -(C 1-5 alkyl)-pyrrolidinyl, -(C 1-5 alkyl)-piperidinyl, -(C 1-5 alkyl)-piperazinyl, -(C 1-5 Alkyl)-morpholinyl, -(C 1-5 alkyl)-thiomorpholinyl, -(C 1-5 alkyl)-imidazolidinyl, -(C 1-5 alkyl)-thiazolidinyl, -(C 1-5 alkyl)-tetrahydrofuranyl, -(C 1-5 Alkyl)-azepanyl, -(C 1-5 alkyl)-diazepanyl, -(C 1-5 Alkyl)-oxazepanyl and -(C 1-5 Any one selected from the group consisting of alkyl)-thiazepanyl; A compound represented by the formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein R3 is unsubstituted or substituted with at least one halogen, C1-C5 alkyl, C1-C5 haloalkyl, -O-C1-C5 haloalkyl, -O-C1-C5 alkyl, -C(=O)-(C1-C5 alkyl), -C(=O)-O-(C1-C5 alkyl), -OH, -NO2, or -NH2.

4. In paragraph 1, R3 is , , , , , , , , , , , , and is one selected from the group consisting of, A compound represented by the formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein R3 is unsubstituted or substituted with at least one halogen, C1-C5 alkyl, C1-C5 haloalkyl, -O-C1-C5 haloalkyl, -O-C1-C5 alkyl, -C(=O)-(C1-C5 alkyl), -C(=O)-O-(C1-C5 alkyl), -OH, -NO2, or -NH2.

5. In paragraph 1, R3 is , , , , , , , , , , , , , , , , and A compound represented by the chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is one selected from the group consisting of:

6. In paragraph 1, R1 and R2 may be the same or different, and are each independently one selected from the group consisting of methyl, ethyl, propyl, and isopropyl, A compound represented by chemical formula 1, wherein n is an integer from 2 to 5, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

7. In paragraph 1, R4 is -O-(C 1-3 A compound represented by the formula 1, wherein the compound is an alkyl) or -OH, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

8. In paragraph 1, A compound represented by the above chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound represented by the above chemical formula 1 is a compound represented by the chemical formula 2-1 or 2-2: [Chemical Formula 2-1] [Chemical Formula 2-2] In the above chemical formula 2-1 or 2-2, R1 and R2 may be the same or different, and each independently represent unsubstituted or substituted C 1-5 It is alkyl; n is an integer from 2 to 10; R3 is -NRaRb, NH-Rc, or unsubstituted or substituted C containing at least one N. 2-8 is heterocycloalkyl; Ra and Rb may be the same or different, and each independently represents -H or unsubstituted or substituted C. 1-5 It is alkyl; Rc is unsubstituted or substituted C 1-5 Alkyl, unsubstituted or substituted C 6-12 Aryl, unsubstituted or substituted -(C 1-5 alkyl)-(C 6-12 Aryl), unsubstituted or substituted C 2-12 Heteroaryl, unsubstituted or substituted -(C 1-5 alkyl)-(C 2-12 heteroaryl), unsubstituted or substituted C 2-8 Heterocycloalkyl, or unsubstituted or substituted -(C 1-5 alkyl)-(C 2-8 heterocycloalkyl); R4 is halogen, C 1-3 Alkyl, -O-(C 1-3 alkyl), -CN, -OH, -NH2, -NO2 or C 1-3 It is haloalkyl.

9. In paragraph 1, The compound represented by the above chemical formula 1 is any one selected from the group consisting of the compounds described below, a compound represented by the chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof: N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(3-hydroxy-4-(phenylcarbamoyl)phenyl)thiophene-2-carboxamide; N-(2-(dimethylamino)ethyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide; N-(4-(dimethylamino)butyl)-5-(3-hydroxy-4-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide; tert-Butyl 4-(4-(5-((3-(dimethylamino)propyl)carbamoyl)thiophen-2-yl)-2-methoxybenzoyl)piperazine-1-carboxylate; N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(piperazine-1-carbonyl)phenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(4-methylpiperazine-1-carbonyl)phenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(4-(4-ethylpiperazine-1-carbonyl)-3-methoxyphenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(4-(4-isopropylpiperazine-1-carbonyl)-3-methoxyphenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(4-(ethylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide; 5-(4-(diethylcarbamoyl)-3-methoxyphenyl)-N-(3-(dimethylamino)propyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(4-(isopropylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(4-(isobutylcarbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(piperidine-1-carbonyl)phenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(pyrrolidine-1-carbonyl)phenyl)thiophene-2-carboxamide; 5-(4-(benzylcarbamoyl)-3-methoxyphenyl)-N-(3-(dimethylamino)propyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(phenethylcarbamoyl)phenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(3-methoxy-4-(pyridin-3-ylcarbamoyl)phenyl)thiophene-2-carboxamide; N-(3-(dimethylamino)propyl)-5-(4-((furan-2-ylmethyl)carbamoyl)-3-methoxyphenyl)thiophene-2-carboxamide; and N-(3-(dimethylamino)propyl)-5-(4-hydroxy-3-(morpholine-4-carbonyl)phenyl)thiophene-2-carboxamide.

10. In paragraph 1, A compound represented by the formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof inhibits the activity of KDM4.

11. A pharmaceutical composition for preventing or treating brain disease, comprising a compound according to any one of claims 1 to 10, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

12. In paragraph 11, The above brain diseases include senile dementia including Alzheimer's disease, vascular dementia, mild cognitive impairment, Parkinson's disease, Huntington's disease, cerebral amyloid angiopathy, Down syndrome, stroke, systemic amyloid disease, amyotrophic lateral sclerosis, multiple sclerosis, stroke, encephalitis, spinocerebellar atrophy, Tourette's syndrome, Friedrich's ataxia, Lewy body dementia, progressive supranuclear palsy, frontotemporal dementia, and Wernicke-Korsakoff's disease. A pharmaceutical composition for preventing or treating a brain disease, wherein the composition is any one selected from the group consisting of Wernicke-Korsakoff's syndrome.

13. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

14. A method for treating a brain disease, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 10, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

15. In paragraph 14, The above brain diseases include senile dementia including Alzheimer's disease, vascular dementia, mild cognitive impairment, Parkinson's disease, Huntington's disease, cerebral amyloid angiopathy, Down syndrome, stroke, systemic amyloid disease, amyotrophic lateral sclerosis, multiple sclerosis, stroke, encephalitis, spinocerebellar atrophy, Tourette's syndrome, Friedrich's ataxia, Lewy body dementia, progressive supranuclear palsy, frontotemporal dementia, and Wernicke-Korsakoff's disease. A method, wherein the method comprises any one selected from the group consisting of WernickeKorsakoff's syndrome.

16. Use of a compound according to any one of claims 1 to 10, an optical isomer thereof, or a pharmaceutically acceptable salt thereof for the manufacture of a drug for treating brain diseases.

17. In paragraph 16, The above brain diseases include senile dementia including Alzheimer's disease, vascular dementia, mild cognitive impairment, Parkinson's disease, Huntington's disease, cerebral amyloid angiopathy, Down syndrome, stroke, systemic amyloid disease, amyotrophic lateral sclerosis, multiple sclerosis, stroke, encephalitis, spinocerebellar atrophy, Tourette's syndrome, Friedrich's ataxia, Lewy body dementia, progressive supranuclear palsy, frontotemporal dementia, and Wernicke-Korsakoff's disease. Any one selected from the group consisting of WernickeKorsakoff's syndrome.

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