Nitrogen-containing saturated heterocyclic derivative

Abstract

The present invention relates to a drug for treating or preventing a central nervous system disease whose cause is an abnormal aggregation of proteins in the brain, comprising, as an active ingredient, a compound of formula (1) (wherein R1 and R2 are hydrogen or the like, R3 and R4 are hydrogen, C1-6 alkyl or the like, R5 is halogen, C1-6 alkyl or the like, R6 is hydrogen, halogen or the like, X is oxygen or the like, Y is carbon or the like, and a pharmaceutically acceptable salt thereof, in the formula, m and n are integers of 0, 1 or the like, r and s are 0, 1, 2 or the like, and Hy is a pyridine ring or the like, which has an effect of inhibiting or reducing the accumulation of abnormal proteins in the brain. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a nitrogen-containing saturated heterocyclic derivative or a pharmaceutically acceptable salt thereof, which has the effect of inhibiting or reducing the accumulation of abnormal protein aggregation in the brain, and a drug containing the derivative as an active ingredient for treating or preventing central nervous system diseases whose causes are related to abnormal protein aggregation in the brain. In addition, the present invention provides a method for reproducing Parkinson's disease pathology with neurospheroids, and a method for evaluating the amount of alpha-synuclein aggregates using the reproducing method. Background Art

[0002] Neurodegenerative diseases mainly include Alzheimer's disease, Parkinson's disease, Huntington's disease and amyotrophic lateral sclerosis, which are believed to be caused by the formation of abnormal aggregated proteins in the patient's brain. The aggregates exhibit neurotoxicity, thus causing the occurrence and development of the disease.

[0003] The constituent proteins of aggregates vary depending on the disease, and α-synuclein has been reported as the main component of aggregates that cause Parkinson's disease. It has been reported that abnormally aggregated α-synuclein exhibits toxicity and that aggregated α-synuclein spreads between cells.

[0004] As a drug for treating Parkinson's disease, administration of levodopa, a dopamine precursor, can be used as a symptomatic treatment, but there is currently no established fundamental treatment. In recent years, the development of some disease-modifying drugs for Parkinson's disease has been actively pursued, but there are currently no reports on drugs undergoing clinical trials that strongly inhibit or reduce α-synuclein aggregation.

[0005] α-synuclein aggregates are believed to be the pathogenic background of Lewy body diseases including Parkinson's disease (Lewy body dementia, multiple system atrophy, Gaucher disease, infantile neuroaxonal dystrophy, etc.). Therefore, drugs that inhibit or reduce the accumulation of α-synuclein aggregates are expected to show therapeutic effects on these diseases.

[0006] Until now, no in vitro evaluation system that reproduces endogenous α-synuclein aggregates in neurons has been reported, and many evaluation systems for α-synuclein pathology are expressed by the amount of phosphorylated α-synuclein increased by the addition of in vitro synthesized α-synuclein oligomers. After all, it is currently impossible to evaluate the effects of inhibiting the accumulation of α-synuclein aggregates or reducing the accumulated α-synuclein aggregates.

[0007] Until now, NPT200-11 (Neuropore) and Anle138b (MODAG) have been reported as drugs that can inhibit the formation of α-synuclein aggregates. However, the effects of these drugs on the ability to inhibit the formation of aggregates when α-synuclein is artificially aggregated in vitro have been evaluated (Patent Documents 1 and 2).

[0008] It has also been reported that (4aR,8aS)-hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one derivatives such as (4aR,8aS)-6-{4-[5-(trifluoromethyl)pyridin-3-yl]piperidine-1-carbonyl}hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one and (4aR,8aS)-6-[4-(5-ethylpyridin-3-yl)piperidine-1-carbonyl]hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one have the effect of inhibiting monoacylglycerol lipase (MAGL) and are effective for neuroinflammation, neurodegenerative diseases, etc. (Patent Document 3). In addition, (azetidin-1-yl)(phenyl)methanone derivatives such as 2-chloro-3-{3-[6-(trifluoromethyl)pyridin-3-yl]azetidine-1-carbonyl}-4-[(1,1,1-trifluoropropane-2-yl)oxy]benzonitrile and 2-methoxy-3-{3-[6-(trifluoromethyl)pyridin-3-yl]azetidine-1-carbonyl}-4-[(1,1,1-trifluoropropane-2-yl)oxy]benzonitrile have been reported to have the effect of inhibiting glycine transporter 1 (GlyT1) and are effective for neurodegenerative diseases, etc. (Patent Document 4).

[0009] However, all these compounds are different from the nitrogen-containing saturated heterocyclic derivatives of the present invention. Moreover, these documents neither disclose nor indicate any information about the nitrogen-containing saturated heterocyclic derivatives of the present invention. In addition, they do not indicate the effect of inhibiting or reducing the accumulation of abnormal protein aggregation in the brain.

[0010] Prior art

[0011] (Patent References)

[0012] [Patent Document 1] WO 2011 / 084642

[0013] [Patent Document 2] WO 2010 / 000372

[0014] [Patent Document 3] WO 2019 / 180185

[0015] [Patent Document 4] WO 2016 / 073420 Summary of the invention

[0016] (Technical Issues)

[0017] The object of the present invention is to provide a compound or a pharmaceutically acceptable salt thereof for preventing or treating a central nervous system disease, characterized by inhibiting or reducing the accumulation of abnormal protein aggregation in the brain, and a composition comprising the compound. In addition, the object of the present invention is also to provide a method for reproducing Parkinson's disease pathology with a neurospheroid, and a method for evaluating the amount of alpha-synuclein aggregates using a reproducing method.

[0018] (Solution to the problem)

[0019] The present inventors have conducted extensive research to achieve the above purpose, and then found that the following formula (1) compound or its pharmaceutically acceptable salt (as the case may be referred to as "the present compound") has an effect of inhibiting or reducing the accumulation of abnormal aggregation of proteins in the brain, and a method for reproducing Parkinson's disease pathology with neurospheroids, and a method for evaluating the amount of α-synuclein aggregates using a reproducing method. Based on these findings, the present invention has been achieved. The present invention is as follows.

[0020] (Item 1)

[0021] A compound of formula (1) or a pharmaceutically acceptable salt thereof:

[0022]

[0023] in

[0024] X is oxygen or NR 7 ,

[0025] R 7 is hydrogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkyl or cyclopropyl,

[0026] Y is CH or nitrogen,

[0027] m is 0, 1, or 2,

[0028] n is 0 or 1,

[0029] r is 0, 1, 2, 3, or 4,

[0030] s is 0, 1, or 2,

[0031] Provided that when s is 0, then Y is CH, and r is 1, 2, 3 or 4,

[0032] When s is 1, then Y is CH, and r is 0, 1, 2 or 3, and

[0033] When s is 2, then r is 1 or 2.

[0034] R1 is hydrogen, halogen, methyl or hydroxy,

[0035] R 2 is hydrogen, halogen, methyl or hydroxy,

[0036] R 3 is hydrogen or C 1-3 alkyl,

[0037] R 4 is hydrogen or C 1-3 alkyl,

[0038] Or, R 3 and R 4 may optionally together form a bridged methylene or a bridged ethylene,

[0039] R 5 is halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy,

[0040] R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and

[0041] Hy is a pyridine ring, a pyridazine ring, a pyrimidine ring or a pyrazine ring,

[0042] The premise is

[0043] (I) When with R 5 and R 6 Hy is 5-fluoropyridin-2-yl, and m and n are 1, then r is 0, and s is 1,

[0044] (II) When R 5 and R 6 When Hy is 6-methoxypyridin-3-yl, m and n are 1, and X is oxygen, then r is 0, and s is 1,

[0045] (III) When R 5 and R 6 Hy is 5-methoxypyridin-2-yl, m and n are 1, and X is NR 7 When , r is 0, and s is 1,

[0046] (IV) When R 5 is methyl, and R 6 When it is hydrogen, r is 0, and s is 1,

[0047] but,

[0048] (I') with R 5 and R 6 Hy is not 4,6-dimethylpyrimidin-2-yl or 5-bromopyrimidin-2-yl,

[0049] (II') When with R 5 and R 6 When Hy is 5-chloropyridin-2-yl, m and n are 1, and R 1 and R 2 Neither of them is hydrogen,

[0050] Provided that the following compounds are not included

[0051] (III') (1-isopropylpiperidin-4-yl){3-(2-methoxypyridin-3-yl)pyrrolidin-1-yl}methanone, and

[0052] (IV') (3-ethoxyoxetan-3-yl)[3-{4-(trifluoromethyl)pyrimidin-2-yl}pyrrolidin-1-yl]methanone.

[0053] (Item 2)

[0054] The compound of Item 1 or a pharmaceutically acceptable salt thereof, wherein m is 1, and n is 1.

[0055] (Item 3)

[0056] The compound of item 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 are independently hydrogen, methyl or fluoro.

[0057] (Item 4)

[0058] The compound of item 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 For hydrogen.

[0059] (Item 5)

[0060] The compound or pharmaceutically acceptable salt thereof according to any one of items 1 to 4, wherein X is oxygen, NH or NMe.

[0061] (Item 6)

[0062] The compound of any one of items 1 to 5 or a pharmaceutically acceptable salt thereof, wherein R 3 For hydrogen.

[0063] (Item 7)

[0064] The compound of any one of items 1 to 6 or a pharmaceutically acceptable salt thereof, wherein R 4 It is methyl or ethyl.

[0065] (Item 8)

[0066] The compound of any one of items 1 to 7 or a pharmaceutically acceptable salt thereof, wherein Y is CH.

[0067] (Item 9)

[0068] The compound according to any one of items 1 to 8, or a pharmaceutically acceptable salt thereof, wherein s is 1.

[0069] (Item 10)

[0070] The compound of any one of items 1 to 9, or a pharmaceutically acceptable salt thereof, wherein r is 0, and s is 1.

[0071] (Item 11)

[0072] The compound of item 1 or a pharmaceutically acceptable salt thereof, wherein the compound is represented by formula (2):

[0073]

[0074] in

[0075] X is oxygen, NH or NMe,

[0076] R 4 is methyl or ethyl,

[0077] R 5 is halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy,

[0078] R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and

[0079] Hy is a pyridine ring, a pyridazine ring, a pyrimidine ring or a pyrazine ring,

[0080] The premise is that it has R 5 and R 6 The Hy of is not 5-chloropyridin-2-yl.

[0081] (Item 12)

[0082] The compound or pharmaceutically acceptable salt thereof according to any one of items 1 to 11, wherein Hy is a pyridine ring.

[0083] (Item 13)

[0084] The compound of any one of items 1 to 12 or a pharmaceutically acceptable salt thereof, wherein R 5 It is trifluoromethyl.

[0085] (Item 14)

[0086] The compound or pharmaceutically acceptable salt thereof according to any one of items 1 to 13, wherein Hy is pyridin-3-yl.

[0087] (Item 15)

[0088] The compound according to any one of items 1 to 14, or a pharmaceutically acceptable salt thereof, wherein X is oxygen.

[0089] (Item 16)

[0090] The compound of any one of items 1 to 15 or a pharmaceutically acceptable salt thereof, wherein R 4 It is methyl.

[0091] (Item 17)

[0092] The compound or pharmaceutically acceptable salt thereof according to any one of items 1 to 14, wherein X is NH or NMe.

[0093] (Item 18)

[0094] The compound according to any one of items 1 to 14, or a pharmaceutically acceptable salt thereof, wherein X is NMe.

[0095] (Item 19)

[0096] The compound of item 1 or a pharmaceutically acceptable salt thereof is selected from the following compounds:

[0097] (3-methyloxetan-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 1),

[0098] (3-methyloxetan-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 2),

[0099] (3-methyloxetan-3-yl){4-[4-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 3),

[0100] (3-methyloxetan-3-yl){4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 4),

[0101] {4-[5-fluoro-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}(3-methyloxetane-3-yl)methanone (Example 29),

[0102] (3-methyloxetan-3-yl){4-[4-methyl-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 31),

[0103] (1,3-dimethylazetidin-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 41),

[0104] (1,3-dimethylazetidin-3-yl){4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 42),

[0105] (1,3-dimethylazetidin-3-yl){4-[4-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 43), and

[0106] (1,3-Dimethylazetidin-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 44).

[0107] (Item 20)

[0108] The compound of item 1 or a pharmaceutically acceptable salt thereof is selected from the following compounds:

[0109] (3-methyloxetan-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 1),

[0110] (3-methyloxetan-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 2),

[0111] {4-[5-fluoro-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}(3-methyloxetane-3-yl)methanone (Example 29),

[0112] (3-methyloxetan-3-yl){4-[4-methyl-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 31),

[0113] (1,3-dimethylazetidin-3-yl){4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 42), and

[0114] (1,3-Dimethylazetidin-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 44).

[0115] (Item 21)

[0116] A medicament comprising the compound according to any one of items 1 to 20 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0117] (Item 22)

[0118] A drug for treating or preventing a central nervous system disease the cause of which is associated with abnormal aggregation of proteins in the brain, comprising a compound according to any one of items 1 to 20 or a pharmaceutically acceptable salt thereof.

[0119] (Item 23)

[0120] The drug of item 22, wherein the central nervous system disease is associated with tau, alpha-synuclein, TDP-43 or polyglutamine.

[0121] (Item 24)

[0122] The drug of item 22, wherein the central nervous system disorder is Alzheimer's disease, frontotemporal lobar degeneration, Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Gaucher disease, infantile axonal dystrophy, amyotrophic lateral sclerosis, Huntington's disease or spinocerebellar ataxia.

[0123] (Item 25)

[0124] The drug of item 22, wherein the central nervous system disease is associated with alpha-synuclein.

[0125] (Item 26)

[0126] A drug according to item 22, wherein the central nervous system disorder is Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Gaucher disease or infantile neuroaxonal dystrophy.

[0127] (Item 27)

[0128] A method for treating or preventing a central nervous system disease the cause of which is associated with abnormal aggregation of proteins in the brain, comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to any one of items 1 to 20 or a pharmaceutically acceptable salt thereof.

[0129] (Item 28)

[0130] Use of a compound according to any one of items 1 to 20 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing a central nervous system disease the cause of which is associated with abnormal aggregation of proteins in the brain.

[0131] (Item 29)

[0132] A compound according to any one of items 1 to 20 or a pharmaceutically acceptable salt thereof for use in treating or preventing a central nervous system disease the cause of which is associated with abnormal aggregation of a protein in the brain.

[0133] (Item 30)

[0134] A medicament for treating or preventing a central nervous system disease whose cause is associated with abnormal aggregation of proteins in the brain, comprising a compound according to any one of items 1 to 20 or a pharmaceutically acceptable salt thereof in combination with at least one agent selected from the group consisting of L-dopa, a dopamine agonist, a MAO-B inhibitor, a catechol-O-methyltransferase (COMT) inhibitor, an αSyn antibody, and a pharmaceutically acceptable salt thereof.

[0135] (Item 31)

[0136] A medicament for treating or preventing a central nervous system disease the cause of which is associated with abnormal aggregation of proteins in the brain, comprising a compound according to any one of items 1 to 20 or a pharmaceutically acceptable salt thereof in combination with at least one agent selected from the group consisting of L-dopa, a dopamine agonist, a MAO-B inhibitor, a catechol-O-methyltransferase (COMT) inhibitor, an αSyn antibody and a pharmaceutically acceptable salt thereof.

[0137] (Item 32)

[0138] A drug for treating or preventing a central nervous system disease whose cause is associated with abnormal protein aggregation in the brain, comprising as an active ingredient a compound of formula (1) or a pharmaceutically acceptable salt thereof:

[0139]

[0140] in

[0141] X is oxygen or NR 7 ,

[0142] R 7 is hydrogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkyl or cyclopropyl,

[0143] Y is CH or nitrogen,

[0144] m is 0, 1, or 2,

[0145] n is 0 or 1,

[0146] r is 0, 1, 2, 3, or 4,

[0147] s is 0, 1, or 2,

[0148] Provided that when s is 0, then Y is CH, and r is 1, 2, 3 or 4,

[0149] When s is 1, then Y is CH, and r is 0, 1, 2 or 3, and

[0150] When s is 2, then r is 1 or 2.

[0151] R 1 is hydrogen, halogen, methyl or hydroxy,

[0152] R 2 is hydrogen, halogen, methyl or hydroxy,

[0153] R 3 is hydrogen or C 1-3 alkyl,

[0154] R 4 is hydrogen or C 1-3 alkyl,

[0155] Or, R 3 and R 4 may optionally together form a bridged methylene or a bridged ethylene,

[0156] R 5 is hydrogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy,

[0157] R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and

[0158] Hy is a pyridine ring, a pyridazine ring, a pyrimidine ring or a pyrazine ring.

[0159] (Item 33)

[0160] The drug of entry 32, wherein m is 1, and n is 1.

[0161] (Item 34)

[0162] A drug of entry 32 or 33, of which R 1 and R 2 are independently hydrogen, methyl or fluoro.

[0163] (Item 35)

[0164] A drug of entry 32 or 33, of which R 1 and R 2 For hydrogen.

[0165] (Item 36)

[0166] The drug of any one of items 32 to 35, wherein X is oxygen, NH or NMe.

[0167] (Item 37)

[0168] Any drug of items 32-36, wherein R 3 For hydrogen.

[0169] (Item 38)

[0170] Any drug of items 32-37, wherein R 4 It is methyl or ethyl.

[0171] (Item 39)

[0172] The drug of any one of items 32 to 38, wherein Y is CH.

[0173] (Item 40)

[0174] A drug according to any of items 32-39, wherein s is 1.

[0175] (Item 41)

[0176] The drug of any one of items 32-40, wherein r is 0, and s is 1.

[0177] (Item 42)

[0178] The drug of item 32, wherein the compound is represented by formula (2):

[0179]

[0180] in

[0181] X is oxygen, NH or NMe,

[0182] R 4 is methyl or ethyl,

[0183] R 5 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy,

[0184] R 6is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and

[0185] Hy is a pyridine ring, a pyridazine ring, a pyrimidine ring or a pyrazine ring,

[0186] (Item 43)

[0187] The drug of any one of items 32 to 42, wherein Hy is a pyridine ring.

[0188] (Item 44)

[0189] Any drug of items 32-43, wherein R 5 It is trifluoromethyl.

[0190] (Item 45)

[0191] The drug of any one of items 32 to 44, wherein Hy is pyridin-3-yl.

[0192] (Item 46)

[0193] The drug of any one of items 32 to 45, wherein X is oxygen.

[0194] (Item 47)

[0195] Any drug of items 32-46, wherein R 4 It is methyl.

[0196] (Item 48)

[0197] The drug of any one of items 32 to 45, wherein X is NH or NMe.

[0198] (Item 49)

[0199] The drug of any one of items 32 to 45, wherein X is NMe.

[0200] (Item 50)

[0201] The drug of any one of items 32-49, wherein the central nervous system disease is related to tau, alpha-synuclein, TDP-43 or polyglutamine.

[0202] (Item 51)

[0203] A medicine as defined in any of items 32 to 49, wherein the central nervous system disorder is Alzheimer's disease, frontotemporal lobar degeneration, Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Gaucher disease, infantile axonal dystrophy, amyotrophic lateral sclerosis, Huntington's disease or spinocerebellar ataxia.

[0204] (Item 52)

[0205] The drug of any one of items 32 to 49, wherein the central nervous system disease is associated with alpha-synuclein.

[0206] (Item 53)

[0207] A medicine as defined in any of items 32 to 49, wherein the central nervous system disorder is Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Gaucher disease or infantile neuroaxonal dystrophy.

[0208] (Item 54)

[0209] A method for treating or preventing a central nervous system disease the cause of which is associated with abnormal aggregation of proteins in the brain, comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to any one of items 32 to 49 or a pharmaceutically acceptable salt thereof.

[0210] (Item 55)

[0211] Use of a compound according to any one of items 32 to 49 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing a central nervous system disease the cause of which is associated with abnormal aggregation of proteins in the brain.

[0212] (Item 56)

[0213] A compound according to any one of items 32 to 49 or a pharmaceutically acceptable salt thereof for use in treating or preventing a central nervous system disease the cause of which is associated with abnormal aggregation of a protein in the brain.

[0214] (Item 57)

[0215] A medicament for treating or preventing a central nervous system disease the cause of which is associated with abnormal aggregation of proteins in the brain, comprising a compound according to any one of items 32 to 49 or a pharmaceutically acceptable salt thereof in combination with at least one agent selected from the group consisting of L-dopa, a dopamine agonist, a MAO-B inhibitor, a catechol-O-methyltransferase (COMT) inhibitor, an αSyn antibody and a pharmaceutically acceptable salt thereof.

[0216] (Item 58)

[0217] A medicament comprising a compound according to any one of items 32 to 49 or a pharmaceutically acceptable salt thereof for use in treating or preventing a central nervous system disease the cause of which is associated with abnormal aggregation of proteins in the brain, in combination with at least one agent selected from the group consisting of L-dopa, a dopamine agonist, a MAO-B inhibitor, a catechol-O-methyltransferase (COMT) inhibitor, an αSyn antibody and a pharmaceutically acceptable salt thereof.

[0218] (Item 59)

[0219] A method for reproducing Parkinson's disease pathology with neurospheroids prepared by three-dimensionally culturing human iPS cells with gene mutations associated with synucleinopathy, comprising step (I);

[0220] Step (I): Measuring the amount of α-synuclein aggregates in neurospheroids.

[0221] (Item 60)

[0222] A method for evaluating a drug having an effect of inhibiting / reducing the accumulation of α-synuclein aggregates in Parkinson's disease pathology using neurospheroids prepared by three-dimensionally culturing human iPS cells with gene mutations associated with synucleinopathy, comprising step (I);

[0223] Step (I): Measuring the amount of α-synaptophysin aggregates in neurospheroids.

[0224] (Item 61)

[0225] A method for preparing a compound of formula (3) or a pharmaceutically acceptable salt thereof:

[0226]

[0227] in

[0228] R 4 is methyl or ethyl,

[0229] R 5 is trifluoromethyl,

[0230] R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and

[0231] Hy is pyridin-3-yl,

[0232] The steps include:

[0233] (Step 1)

[0234] The compound of formula (4) or its salt:

[0235]

[0236] Where R 5 , R 6 and Hy are defined above,

[0237] Condensation with a compound of formula (5) or a salt thereof:

[0238]

[0239] Where R 4 As defined above, and A is OH or halogen,

[0240] To prepare the compound of formula (3) or a pharmaceutically acceptable salt thereof.

[0241] (Item 62)

[0242] A method for preparing a compound of formula (3) or a pharmaceutically acceptable salt thereof:

[0243]

[0244] in

[0245] R 4 is methyl or ethyl,

[0246] R 5 is trifluoromethyl,

[0247] R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and

[0248] Hy is pyridin-3-yl,

[0249] The method comprises the following steps 1;

[0250] (Step 1)

[0251] The compound of formula (4) or its salt:

[0252]

[0253] Where R 5 , R 6 and Hy are defined above,

[0254] Condensation with a compound of formula (5) or a salt thereof in the presence of 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphaninane-2,4,6-trioxide and triethylamine:

[0255]

[0256] Where R 4 As defined above, and A is OH,

[0257] To prepare the compound of formula (3) or a pharmaceutically acceptable salt thereof.

[0258] (Item 63)

[0259] A method for preparing a compound of formula (6) or a pharmaceutically acceptable salt thereof:

[0260]

[0261] in

[0262] R 4 Methyl,

[0263] R 5 is trifluoromethyl,

[0264] R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and

[0265] Hy is pyridin-3-yl,

[0266] The steps include 1-3:

[0267] (Step 1)

[0268] The compound of formula (4) or its salt:

[0269]

[0270] Where R 5 , R 6 and Hy are defined above,

[0271] Condensation with a compound of formula (7) or a salt thereof:

[0272]

[0273] Where R 4 As defined above, A is OH or halogen, and Pro is an amino protecting group, to prepare a compound of formula (8) or a pharmaceutically acceptable salt thereof:

[0274]

[0275] Where R 4 , R5 , R 6 , Hy and Pro are as defined above,

[0276] (Step 2)

[0277] Deprotecting the amino protecting group in the compound of formula (8) or its salt to prepare the compound of formula (9) or its salt,

[0278]

[0279] Where R 4 , R 5 , R 6 and Hy are defined above,

[0280] (Step 3)

[0281] The compound of formula (9) or a salt thereof is reacted with formaldehyde or an equivalent compound thereof in the presence of a reducing agent to prepare a compound of formula (6) or a pharmaceutically acceptable salt thereof.

[0282] (Item 64)

[0283] A method for preparing a compound of formula (6) or a pharmaceutically acceptable salt thereof:

[0284]

[0285] in

[0286] R 4 Methyl,

[0287] R 5 is trifluoromethyl,

[0288] R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and

[0289] Hy is pyridin-3-yl,

[0290] The steps include 1-3:

[0291] (Step 1)

[0292] The compound of formula (4) or its salt:

[0293]

[0294] Where R 5 , R 6 and Hy are defined above,

[0295] Condensation with a compound of formula (7) or a salt thereof in the presence of 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphaninane-2,4,6-trioxide and triethylamine:

[0296]

[0297] Where R 4 As defined above, Pro is tert-butoxycarbonyl, and A is OH, to prepare a compound of formula (8) or a salt thereof:

[0298]

[0299] Where R 4 , R 5 , R 6 , Hy and Pro are as defined above,

[0300] (Step 2)

[0301] The amino protecting group in the compound of formula (8) or its salt is deprotected with trifluoroacetic acid to prepare the compound of formula (9) or its salt,

[0302]

[0303] Where R 4 , R 5 , R 6 and Hy are defined above,

[0304] (Step 3)

[0305] The compound of formula (9) or a salt thereof is reacted with formaldehyde in the presence of sodium triacetoxyborohydride and acetic acid to prepare the compound of formula (6) or a pharmaceutically acceptable salt thereof.

[0306] (Effects of the Invention)

[0307] The present invention makes it possible to provide a compound of formula (1) or a pharmaceutically acceptable salt thereof. The compound or a pharmaceutically acceptable salt thereof has the effect of inhibiting or reducing the accumulation of abnormal aggregation of proteins in the brain, and can thus be used as a drug for treating or preventing central nervous system diseases whose causes are related to abnormal aggregation of proteins in the brain, and the disease is particularly a neurodegenerative disease associated with alpha-synuclein, such as Parkinson's disease, Lewy body dementia, multiple system atrophy, Gaucher disease and infantile neuroaxonal dystrophy. In addition, the present invention can reproduce spontaneous alpha-synuclein aggregates as the pathology of Parkinson's disease in neurons, and thus the present invention can be used as a method for evaluating a drug having an effect of inhibiting or reducing the accumulation of alpha-synuclein aggregates. BRIEF DESCRIPTION OF THE DRAWINGS

[0309] Figure 1The difference in the amount of aggregates in neurospheres derived from healthy human iPS cells and neurospheres derived from PLA2G6 mutant iPS cells is shown. The vertical axis represents the amount of aggregates in neurospheres, and the horizontal axis represents the number of culture days. The white bars represent the amount of aggregates in neurospheres derived from healthy human iPS cells, and the black bars represent the amount of aggregates in neurospheres derived from PLA2G6 mutant iPS cells.

[0310] Figure 2 The difference in the amount of aggregates in dopamine neurospheres derived from healthy human iPS cells and dopamine neurospheres derived from PLA2G6 mutant iPS cells is shown. The vertical axis represents the amount of aggregates in dopamine neurospheres, and the horizontal axis represents the number of culture days. The white bars represent the amount of aggregates in neurospheres derived from healthy human iPS cells, and the black bars represent the amount of aggregates in neurospheres derived from PLA2G6 mutant iPS cells.

[0311] Figure 3 The figure shows the difference in the amount of tyrosine hydroxylase in dopamine neurospheres derived from healthy human iPS cells and dopamine neurospheres derived from PLA2G6 mutant iPS cells on culture day 26. The vertical axis represents the amount of tyrosine hydroxylase in dopamine neurospheres. The white bar represents the amount of tyrosine hydroxylase in neurospheres derived from healthy human iPS cells, and the black bar represents the amount of tyrosine hydroxylase in neurospheres derived from PLA2G6 mutant iPS cells.

[0312] Figure 4 The difference in the amount of cleaved caspase 3 between dopamine neurospheres derived from healthy human iPS cells and dopamine neurospheres derived from PLA2G6 mutant iPS cells on culture day 40 is shown. The vertical axis represents the amount of cleaved caspase 3 in dopamine neurospheres. The white bar represents the amount of cleaved caspase 3 in neurospheres derived from healthy human iPS cells, and the black bar represents the amount of cleaved caspase 3 in neurospheres derived from PLA2G6 mutant iPS cells.

[0313] Figure 5 The difference in the amount of aggregates in dopamine neurospheres derived from healthy human iPS cells and dopamine neurospheres derived from GBA1 gene homozygous mutation iPS cells is shown. The vertical axis represents the amount of aggregates in dopamine neurospheres, and the horizontal axis represents the number of culture days. The white bars represent the amount of aggregates in neurospheres derived from healthy human iPS cells, and the black bars represent the amount of aggregates in neurospheres derived from GBA1 gene homozygous mutation iPS cells. DETAILED DESCRIPTION

[0314] Hereinafter, the present invention is explained in detail. In this specification, the number of carbon atoms in the definition of "substituent" may be expressed as, for example, "C 1-3 Specifically, the term "C 1-3 "Alkyl" is synonymous with an alkyl group having 1 to 3 carbon atoms.

[0315] The "halogen" includes, for example, fluorine, chlorine, bromine and iodine. It is preferably fluorine or chlorine.

[0316] “C 1-3 "Alkyl" means a straight or branched chain saturated hydrocarbon group having 1 to 3 carbon atoms. 1-3 The alkyl group preferably includes "C 1-2 Alkyl". "C 1-3 The term "alkyl" includes, for example, methyl, ethyl, propyl and isopropyl.

[0317] “C 1-3 The "C" in "alkoxy" 1-3 Alkyl" as above "C 1-3 "C 1-3 "Alkoxy" preferably includes "C 1-2 Alkoxy". "C 1-3 The "alkoxy group" includes, for example, methoxy, ethoxy, propoxy and isopropoxy.

[0318] Preferred X, Y, m, n, r, s, R in the compound of formula (1) are 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 As shown below, the technical scope of the present invention is not limited to the range of compounds listed below.

[0319] If carbon is substitutable, then R in the compound of formula (1) 1 , R 2 , R 3 , R 4 , R 5 and R 6 The group may be substituted on any carbon; if possible, R 1 and R 2 and / or R 3 and R 4 may be substituted on the same carbon; and when Y is CH, R 3 or R 4 It can replace H of CH.

[0320] In the compound of formula (1), the binding site (indicated by the arrow below) of Hy to the nitrogen-containing saturated heterocyclic ring is carbon.

[0321]

[0322] X preferably includes oxygen, NH and NMe, more preferably oxygen and NMe.

[0323] Y preferably comprises CH.

[0324] n preferably comprises 1.

[0325] m preferably includes 1.

[0326] When Y is CH, r is preferably 0, 1 or 2, more preferably 0 or 1, even more preferably 0.

[0327] When Y is nitrogen, r is 1, 2, 3 or 4, preferably 1.

[0328] When Y is CH, s is preferably 0, 1 or 2, more preferably 1 or 2, even more preferably 1. When Y is nitrogen, s is 2.

[0329] R 1 Preferred include hydrogen, methyl and fluorine, more preferably hydrogen.

[0330] R 2 Preferred include hydrogen, methyl and fluorine, more preferably hydrogen.

[0331] R 3 Preferred include hydrogen, methyl and ethyl, more preferably hydrogen.

[0332] R 4 Preferred include hydrogen, methyl and ethyl, more preferably methyl and ethyl, even more preferably methyl.

[0333] In a ring containing X and Y, when R 3 and R 4 When forming a bridged methylene group or a bridged ethylene group together, the structure of the ring containing X and Y includes the structure of the following formula (3). In the following formula (3), the wavy line indicates the binding site to the carbonyl group in formula (1). In the following formula (3), X and Y are as defined in item 1.

[0334]

[0335] R 5 Preferably includes halogen and C optionally substituted with 1 to 3 identical or different halogen atoms. 1-3 Alkyl, more preferably halogen and methyl optionally substituted with 1 to 3 fluorine atoms, even more preferably trifluoromethyl, methyl and fluorine, most preferably trifluoromethyl.

[0336] R 6 Preferably includes hydrogen, halogen and C optionally substituted with 1 to 3 identical or different halogen atoms.1-3 Alkyl, more preferably hydrogen, halogen and methyl optionally substituted with 1 to 3 fluorine atoms, even more preferably hydrogen, halogen and methyl, most preferably hydrogen, fluorine and methyl.

[0337] R 7 Preferably includes hydrogen and C 1-3 Alkyl, more preferably hydrogen and methyl, even more preferably methyl. 7 Another embodiment includes C having 1-5 deuterium atoms. 1-3 alkyl.

[0338] Hy preferably comprises a pyridine ring, more preferably a pyridin-3-yl group of the following formula (4), wherein the arrow indicates the bonding position to the nitrogen-containing saturated ring.

[0339]

[0340] The condensing agent used for the condensation reaction includes 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphaninane-2,4,6-trioxide. Preferably, it includes 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphaninane-2,4,6-trioxide.

[0341] The reducing agent includes sodium borohydride, sodium triacetoxyborohydride and sodium cyanoborohydride. Preferably, it includes sodium cyanoborohydride and sodium triacetoxyborohydride.

[0342] Formaldehyde or its equivalent compound includes formaldehyde, 1,3,5-trioxane and paraformaldehyde. Preferably, it includes formaldehyde.

[0343] Amino protecting groups include tert-butoxycarbonyl and benzyloxycarbonyl, preferably tert-butoxycarbonyl.

[0344] Preferred compounds of formula (1) include the following compounds or pharmaceutically acceptable salts thereof.

[0345] In one embodiment, the compound of formula (1) comprises the following (A).

[0346] (A)

[0347] A compound or a pharmaceutically acceptable salt thereof, wherein

[0348] X is oxygen or NR 7 ,

[0349] R 7 For hydrogen, C 1-3 Alkyl or cyclopropyl,

[0350] Y is CH or nitrogen,

[0351] m is 0, 1, or 2,

[0352] n is 0 or 1,

[0353] r is 0, 1, or 2,

[0354] s is 0, 1, or 2,

[0355] Provided that when s is 0, then Y is CH, and r is 1 or 2,

[0356] When s is 1, then Y is CH, and r is 0, 1 or 2,

[0357] When s is 2, then r is 1 or 2.

[0358] R 1 is hydrogen, halogen, methyl or hydroxy

[0359] R 2 is hydrogen, halogen, methyl or hydroxy,

[0360] R 3 is hydrogen or C 1-3 alkyl,

[0361] R 4 is hydrogen or C 1-3 alkyl,

[0362] Or, R 3 and R 4 may optionally together form a bridged methylene or a bridged ethylene,

[0363] R 5 is halogen or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl,

[0364] R 6 is hydrogen, halogen or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkyl, and

[0365] Hy is a pyridine ring, a pyridazine ring, a pyrimidine ring or a pyrazine ring,

[0366] The premise is

[0367] (I) When with R 5 and R 6 Hy is 5-fluoropyridin-2-yl, and m and n are 1, then r is 0, and s is 1,

[0368] (II) When R 5 is methyl, and R 6 When it is hydrogen, r is 0, and s is 1,

[0369] but,

[0370] (I') with R 5 and R 6 Hy is not 4,6-dimethylpyrimidin-2-yl or 5-bromopyrimidin-2-yl,

[0371] (II') When with R 5 and R 6 When Hy is 5-chloropyridin-2-yl, m and n are 1, and R 1 and R 2 Neither of them is hydrogen,

[0372] Provided that the following compounds are not included:

[0373] (III') (3-ethoxyoxetan-3-yl) [3-{4-(trifluoromethyl)pyrimidin-2-yl}pyrrolidin-1-yl]methanone.

[0374] One embodiment of the compound of formula (1) includes the following (B):

[0375] (B)

[0376] A compound or a pharmaceutically acceptable salt thereof, wherein

[0377] X is oxygen or NR 7 ,

[0378] R 7 For hydrogen, C 1-3 Alkyl or cyclopropyl,

[0379] Y is CH or nitrogen,

[0380] m is 0, 1, or 2,

[0381] n is 0 or 1,

[0382] r is 0, 1, or 2,

[0383] s is 0, 1, or 2,

[0384] Provided that when s is 0, then Y is CH, and r is 1 or 2,

[0385] When s is 1, then Y is CH, and r is 0, 1 or 2,

[0386] When s is 2, then r is 1 or 2.

[0387] R 1 is hydrogen, halogen, methyl or hydroxy,

[0388] R 2 is hydrogen, halogen, methyl or hydroxy,

[0389] R 3 is hydrogen or C 1-3 alkyl,

[0390] R 4 is hydrogen or C 1-3 alkyl,

[0391] Or, R 3 and R 4 may optionally together form a bridged methylene or a bridged ethylene,

[0392] R 5 is halogen or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl,

[0393] R 6 is hydrogen, halogen or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkyl, and

[0394] Hy is a pyridine ring,

[0395] The premise is

[0396] (I) When with R 5 and R 6 Hy is 5-fluoropyridin-2-yl, and m and n are 1, then r is 0, and s is 1,

[0397] (II) When R 5 is methyl, and R 6 When it is hydrogen, r is 0, and s is 1,

[0398] but,

[0399] (I') When with R 5 and R 6 When Hy is 5-chloropyridin-2-yl, m and n are 1, and R 1 and R 2 Neither of them is hydrogen,

[0400] One embodiment of the compound of formula (1) includes the following (C).

[0401] (C)

[0402] A compound or a pharmaceutically acceptable salt thereof, wherein

[0403] X is oxygen or NR 7,

[0404] R 7 For hydrogen, C 1-3 Alkyl or cyclopropyl,

[0405] Y is CH,

[0406] m is 1,

[0407] n is 1,

[0408] r is 0, 1, or 2,

[0409] s is 0, 1, or 2,

[0410] The premise is that when s is 0, then r is 1 or 2,

[0411] When s is 1, then r is 0, 1, or 2, and

[0412] When s is 2, then r is 1 or 2.

[0413] R 1 is hydrogen, methyl or fluorine,

[0414] R 2 is hydrogen, methyl or fluorine,

[0415] R 3 is hydrogen, methyl or ethyl,

[0416] R 4 is hydrogen, methyl or ethyl,

[0417] R 5 is halogen or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl,

[0418] R 6 is hydrogen, halogen or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkyl, and

[0419] Hy is a pyridine ring,

[0420] The premise is

[0421] (I) When with R 5 and R 6 When Hy is 5-fluoropyridin-2-yl, r is 0, and s is 1,

[0422] (II) When R 5 is methyl, and R 6 When it is hydrogen, r is 0, and s is 1,

[0423] but,

[0424] (I') When with R5 and R 6 When Hy is 5-chloropyridin-2-yl, then R 1 and R 2 Neither is hydrogen.

[0425] One embodiment of the compound of formula (1) includes the following (D):

[0426] (D)

[0427] A compound or a pharmaceutically acceptable salt thereof, wherein

[0428] X is oxygen or NR 7 ,

[0429] R 7 is hydrogen or methyl,

[0430] Y is CH,

[0431] m is 1,

[0432] n is 1,

[0433] r is 0 or 1,

[0434] s is 1 or 2,

[0435] The premise is that when s is 2, then r is 1,

[0436] R 1 For hydrogen,

[0437] R 2 For hydrogen,

[0438] R 3 For hydrogen,

[0439] R 4 is methyl or ethyl,

[0440] R 5 is halogen or methyl optionally substituted with 1 to 3 fluorine atoms,

[0441] R 6 is hydrogen, halogen or methyl optionally substituted with 1 to 3 fluorine atoms, and

[0442] Hy is pyridin-3-yl.

[0443] One embodiment of the compound of formula (1) includes the following (E):

[0444] (E)

[0445] A compound of formula (2) or a pharmaceutically acceptable salt thereof:

[0446]

[0447] in

[0448] X is oxygen or NMe,

[0449] R 4 Methyl,

[0450] R 5 is trifluoromethyl, methyl or fluorine,

[0451] R 6 is hydrogen, halogen or methyl, and

[0452] Hy is pyridin-3-yl.

[0453] One embodiment of the compound of formula (1) includes the following compounds or pharmaceutically acceptable salts thereof:

[0454] (3-methyloxetan-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 1),

[0455] (3-methyloxetan-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 2),

[0456] (3-methyloxetan-3-yl){4-[4-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 3),

[0457] (3-methyloxetan-3-yl){4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 4),

[0458] {4-[5-fluoro-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}(3-methyloxetane-3-yl)methanone (Example 29),

[0459] (3-methyloxetan-3-yl){4-[4-methyl-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 31),

[0460] (1,3-dimethylazetidin-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 41),

[0461] (1,3-dimethylazetidin-3-yl){4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 42),

[0462] (1,3-dimethylazetidin-3-yl){4-[4-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 43), or

[0463] (1,3-Dimethylazetidin-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 44).

[0464] One embodiment of the compound of formula (1) includes the following compounds or pharmaceutically acceptable salts thereof:

[0465] (3-methyloxetan-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 1),

[0466] (3-methyloxetan-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 2),

[0467] {4-[5-fluoro-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}(3-methyloxetane-3-yl)methanone (Example 29),

[0468] (3-methyloxetan-3-yl){4-[4-methyl-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 31),

[0469] (1,3-dimethylazetidin-3-yl){4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 42), or

[0470] (1,3-Dimethylazetidin-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone (Example 44).

[0471] "Pharmaceutically acceptable salts" include acid addition salts and base addition salts. For example, acid addition salts include inorganic acid salts such as hydrochlorides, hydrobromides, sulfates, hydroiodides, nitrates and phosphates; and organic acid salts such as citrates, oxalates, phthalates, fumarates, maleates, succinates, malates, acetates, formates, propionates, benzoates, trifluoroacetates, methanesulfonates, benzenesulfonates, p-toluenesulfonates and camphorsulfonates. Base addition salts include inorganic base salts such as sodium salts, potassium salts, calcium salts, magnesium salts, barium salts and aluminum salts; and organic base salts such as trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, tromethamine [tris (hydroxymethyl) methylamine], tert-butylamine, cyclohexylamine, dicyclohexylamine and N,N-dibenzylethylamine. "Pharmaceutically acceptable salts" also include amino acid salts of basic or acidic amino acids such as arginine, lysine, ornithine, aspartic acid and glutamic acid.

[0472] Suitable salts of starting materials and intermediates and acceptable salts of bulk drugs are conventional non-toxic salts. Suitable salts include, for example, acid addition salts, such as organic acid salts (including acetate, trifluoroacetate, maleate, fumarate, citrate, tartrate, methanesulfonate, benzenesulfonate, formates and p-toluenesulfonate) and inorganic acid salts (including hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate and phosphate); salts with amino acids (including arginine, aspartic acid and glutamic acid); alkali metal salts (including sodium and potassium salts); alkaline earth metal salts (including calcium and magnesium salts); ammonium salts; organic base salts (including trimethylamine salts, triethylamine salts, pyridine salts, picolinates, dicyclohexylamine salts and N,N'-dibenzylethylenediamine salts); and other salts that can be optionally selected by those skilled in the art.

[0473] If the compound of the present invention should be obtained as its salt, when the present compound is obtained as a salt, it can be purified without further reaction, and when obtained in a free form, it can be dissolved or suspended in an appropriate organic solvent, and an acid or base can be added thereto by a common method to form a salt.

[0474] In the present invention, the compound of formula (1) includes deuterated compounds, wherein any one or more of the compounds of formula (1) 1 H by 2 H(D) is substituted. The present invention includes compounds of formula (1) or pharmaceutically acceptable salts thereof. The compounds of the present invention may exist in the form of hydrates and / or solvates with various solvents including ethanolates, and these hydrates and / or solvates are included in the compounds of the present invention. In addition, the present invention includes all tautomers of compound (1), all possible stereoisomers thereof, crystalline forms of various states thereof, and mixtures thereof.

[0475] The present compound (1) includes optical isomers based on optically active centers, atropisomers based on axial or planar chirality caused by restricted intramolecular rotation, and all other isomers that may exist as stereoisomers, tautomers and geometric isomers, and mixtures thereof.

[0476] In particular, each optical isomer and atropisomer can be obtained as a racemate, or as an optically active substance when an optically active starting material or intermediate is used. If necessary, in the appropriate step of the above preparation method, the racemate of the corresponding starting material, intermediate or final product can also be physically or chemically resolved into optical enantiomers by known separation methods, such as methods using optically active columns and fractional crystallization methods. These methods for resolving enantiomers include diastereomeric methods, in which, for example, a racemate is reacted with an optically active resolving agent to synthesize two diastereomers, which are resolved by different physical properties by fractional crystallization or similar methods.

[0477] Hereinafter, some examples are shown to explain the method for preparing the compound of formula (1) in the present invention, but the method of the present invention should not be limited thereto.

[0478] Preparation method

[0479] The compound of the present invention can be synthesized according to each of the production methods shown below or a combination thereof with known synthesis methods.

[0480] Each compound in the following schemes may exist as a salt thereof, wherein the salts include, for example, the "pharmaceutically acceptable salts" described above as salts of the compound of formula (1). The following schemes are disclosed only as examples, and thus the present compounds may be optionally prepared by different methods based on the knowledge of those skilled in the art of synthetic organic chemistry.

[0481] In each of the preparation methods described below, even if the use of a protecting group is not explicitly described, a protecting group may be used as necessary. Also, the protecting group may be deprotected after the reaction has been completed or a series of reactions have been performed to obtain a desired compound.

[0482] Introduction and removal of the protecting group can be carried out by a method generally used in organic synthetic chemistry, for example, the method described in TW Greene and PGM Wuts, "Protective Groups in Organic Synthesis", 3rd Ed., John Wiley and Sons, inc., New York (1999), or a method similar thereto.

[0483] Examples of the amino protecting group include, for example, tert-butoxycarbonyl, benzyloxycarbonyl, p-toluenesulfonyl, o-nitrobenzenesulfonyl, 4-methoxybenzyl, 2,4-dimethoxybenzyl and the like.

[0484] Preparation method 1

[0485] Among the compounds of formula (1), the compound of the following formula (1a) can be produced, for example, by the following production method.

[0486]

[0487] Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , m, n, r, s, Y and Hy are as defined in item 1; and A is halogen or OH.

[0488] (Step 1-1: Preparation step of compound (1a))

[0489] In the presence of any condensing agent and / or any base, or in the absence of them, by reacting compound (1-1) with compound (1-2) in a suitable inert solvent, compound (1a) can be prepared. As compound (1-1), commercially available compounds can be used, or products prepared by known methods (such as WO 2014 / 192868) can be used. Alternatively, the product prepared in the preparation method 3-6 shown below can be used as compound (1-1). As compound (1-2), commercially available compounds can be used, or products prepared by known methods (such as WO2016 / 004272) can be used. The alkali used in this step can be appropriately selected from the alkalis exemplified below, including, for example, sodium hydride, triethylamine, diisopropylethylamine and sodium carbonate. The condensing agent used in this step can be selected from various condensing agents commonly used in organic synthesis reactions, including, for example, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphorus heterocyclohexane-2,4,6-trioxide. The solvent used in this step can be appropriately selected from the solvents exemplified below, including, for example, DMF, THF, dichloromethane, chloroform and ethyl acetate. The reaction time in this step is generally 5 minutes to 72 hours, preferably 30 minutes to 24 hours. The reaction temperature in this step is generally -78°C to 200°C, preferably -78°C to 80°C.

[0490] Preparation method 2

[0491] Among the compounds of formula (1), compounds of the following formula (1b) and formula (1c) can be produced, for example, by the following production method.

[0492]

[0493] Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , m, n, r, s, Y and Hy are as defined in item 1; R 8 is C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or cyclopropyl; A is halogen or OH; and Pro is an amino protecting group.

[0494] (Step 2-1: Preparation step of compound (1c))

[0495] According to the method shown in step 1-1, compound (1c) can be prepared by reacting compound (1-1) with compound (2-1). As compound (2-1), a commercially available compound can be used, or a product prepared by a known method (eg, WO 2010 / 026096) can be used.

[0496] (Step 2-2: Preparation step of compound (2-3))

[0497] According to the method shown in step 1-1, compound (2-3) can be prepared by reacting compound (1-1) with compound (2-2). As compound (2-2), a commercially available compound can be used, or a product prepared by a known method (eg, WO2008 / 085117) can be used.

[0498] (Step 2-3: Preparation step of compound (1b))

[0499] Compound (1b) can be prepared by deprotecting the protecting group Pro in compound (2-3) according to a known method (e.g., the method described in Protective Group in Organic Synthesis 3rd Ed. (described by Theodora W. Green, Peter GM Wuts, published by John Wiley & Sons Inc, 1999)). The amino protecting group Pro includes, for example, tert-butoxycarbonyl and benzyloxycarbonyl.

[0500] (Step 2-4: Preparation step of compound (1c))

[0501] The compound (1b) can be prepared by reacting the compound (1b) with its corresponding R 8 The aldehyde compound, ketone compound or ketone equivalent compound is reacted in a suitable inert solvent in the presence of a reducing agent to prepare compound (1c). The reducing agent used in this step can be selected from various reducing agents commonly used in organic synthesis reactions, including, for example, sodium borohydride, sodium triacetoxyborohydride and sodium cyanoborohydride. The solvent used in this step can be appropriately selected from the solvents exemplified below, including, for example, toluene, THF, dichloromethane and methanol. The reaction time in this step is generally 5 minutes to 48 hours, preferably 1 hour to 24 hours. The reaction temperature in this step is generally -78°C to 100°C, preferably 0°C to 80°C.

[0502] Alternatively, the compound (1b) may be reacted with the compound (1b) corresponding to R 8 Any alkyl halide or alkyl sulfonate of is reacted in the presence of a base in a suitable inert solvent to prepare compound (1c). The base used in this step can be appropriately selected from the bases exemplified below, including, for example, potassium carbonate, cesium carbonate, sodium hydride and lithium diisopropylamide. The solvent used in this step can be appropriately selected from the solvents exemplified below, including, for example, DMF, dimethyl sulfoxide, THF and 1,4-dioxane. The reaction time in this step is generally 5 minutes to 48 hours, preferably 1 hour to 24 hours. The reaction temperature in this step is generally -78°C to 100°C, preferably 0°C to 80°C.

[0503] Preparation method 3

[0504] The compound of the following formula (1-1) can be produced, for example, by the following production method.

[0505]

[0506] Where R 1 , R 2 , R 5 , R 6 , m, n and Hy are as defined in item 1; W 1 and W 2 is halogen; and Pro is an amino protecting group.

[0507] (Step 3-1: Preparation step of compound (3-3))

[0508] Compound (3-3) can be prepared by reacting compound (3-1) with compound (3-2) in a suitable inert solvent in the presence of a zinc and palladium catalyst. As compound (3-1), a commercially available compound can be used, or a product prepared by a known method (e.g., WO 2008 / 147831) can be used. As compound (3-2), a commercially available compound can be used, or a product prepared by a known method (e.g., Bioorganic & Medicinal Chemistry Letters (2006), 16 (17), 4528-4532) can be used. Halogen as W 1 and W 2 The amino protecting group Pro includes, for example, tert-butyloxycarbonyl and benzyloxycarbonyl. The palladium catalyst used in this step can be selected from various conventional palladium catalysts, including, for example, tetrakis(triphenylphosphine)palladium(0). The solvent used in this step can be appropriately selected from the solvents exemplified below, including, for example, DMF and dimethylacetamide. The reaction time in this step is generally 5 minutes to 48 hours, preferably 1 hour to 24 hours. The reaction temperature in this step is generally 0°C to 100°C, preferably 0°C to 80°C.

[0509] (Step 3-2: Preparation step of compound (1-1))

[0510] According to the method shown in step 2-3, compound (1-1) can be produced from compound (3-3).

[0511] Preparation method 4

[0512] Among the compounds of formula (1-1), the compound of the following formula (1-1a) can be produced, for example, by the following production method.

[0513]

[0514] Where R 1 , R 2 , R 5 , R 6 , n and Hy are as defined in item 1; W 2 is halogen; Pro is an amino protecting group; T is boronic acid or boronic ester.

[0515] (Step 4-1: Preparation step of compound (4-2))

[0516] Compound (4-2) can be prepared by reacting compound (4-1) with compound (3-2) in a suitable inert solvent in the presence of a palladium catalyst. This step can be carried out in the presence of a base and / or a phosphorus ligand as appropriate. As compound (4-1), a commercially available compound can be used, or a product prepared by a known method (e.g., WO 2019 / 163865) can be used. The amino protecting group Pro includes, for example, tert-butyloxycarbonyl and benzyloxycarbonyl. The palladium catalyst used in this step can be selected from various conventionally used palladium catalysts, including, for example, tetrakis(triphenylphosphine)palladium(0). The base used in this step can be appropriately selected from the bases exemplified below, including, for example, potassium carbonate and cesium carbonate. The phosphorus ligand used in this step can be selected from various phosphorus ligands commonly used in organic synthesis reactions, including, for example, triphenylphosphine and bis(diphenylphosphine)methane. The solvent used in this step can be appropriately selected from the solvents exemplified below, including, for example, 1,4-dioxane, tetrahydrofuran, water, and mixtures thereof. The reaction temperature in this step is generally 0° C.-200° C., preferably 20° C.-150° C., and the reaction may be carried out under microwave irradiation as appropriate. The reaction time in this step may vary according to the reaction temperature, palladium catalyst, material and solvent to be used, but is generally 5 minutes-72 hours, preferably 1 hour-24 hours.

[0517] (Step 4-2: Preparation step of compound (4-3))

[0518] Compound (4-3) can be prepared by reacting compound (4-2) in a suitable inert solvent in the presence of a catalyst under a hydrogen atmosphere. The solvent used in this step can be appropriately selected from the solvents exemplified below, including, for example, methanol, ethanol, chloroform, and mixtures thereof. The catalyst used in this step can be selected from various catalysts commonly used in hydrogenation reactions, including, for example, palladium carbon and palladium hydroxide. The reaction time in this step is generally 5 minutes to 48 hours, preferably 1 hour to 24 hours. The reaction temperature in this step is generally 0°C to 100°C, preferably 0°C to 40°C.

[0519] (Step 4-3: Preparation step of compound (1-1a))

[0520] According to the method shown in step 2-3, compound (1-1a) can be produced from compound (4-3).

[0521] Preparation method 5

[0522] Among the compounds of formula (1-1), the compound of the following formula (1-1b) can be produced, for example, by the following production method.

[0523]

[0524] Where R2 , R 5 , R 6 , m, n and Hy are as defined in item 1; W 2 is halogen; and Pro is an amino protecting group.

[0525] (Step 5-1: Preparation step of compound (5-2))

[0526] Compound (5-2) can be prepared by reacting compound (3-2) with compound (5-1) in a suitable inert solvent in the presence of alkyl lithium. As compound (5-1), commercially available compounds can be used, or products prepared by known methods (e.g., WO 2005 / 058888) can be used. The amino protecting group Pro includes, for example, tert-butyloxycarbonyl and benzyloxycarbonyl. The alkyl lithium used in this step can be selected from various conventionally used alkyl lithiums, including, for example, butyl lithium. The solvent used in this step can be appropriately selected from the solvents exemplified below, including, for example, tetrahydrofuran and ether. The reaction time in this step is generally 5 minutes to 48 hours, preferably 1 hour to 24 hours. The reaction temperature in this step is generally -78°C to 100°C, preferably -78°C to 40°C.

[0527] (Step 5-2: Preparation step of compound (1-1b))

[0528] According to the method shown in step 2-3, compound (1-1b) can be produced from compound (5-2).

[0529] Preparation method 6

[0530] Among the compounds of formula (1-1), the compound of the following formula (1-1c) can be produced, for example, by the following production method.

[0531]

[0532] Where R 2 , R 5 , R 6 , m, n and Hy are as defined in Item 1; Q is halogen; and Pro is an amino protecting group.

[0533] (Step 6-1: Step for preparing compound (6-1))

[0534] Compound (6-1) can be prepared by reacting compound (5-2) in an inert solvent in the presence of a halogenating agent. Amino protecting group Pro includes, for example, tert-butoxycarbonyl and benzyloxycarbonyl. Halogen as Q includes fluorine and chlorine. The halogenating agent used in this step can be selected from various conventionally used halogenating agents, including, for example, (diethylamino) sulfur trifluoride, bis(2-methoxyethyl) aminosulfur trifluoride and phosphorus oxychloride. The solvent used in this step can be appropriately selected from the solvents exemplified below, including, for example, dichloromethane, chloroform, 1,4-dioxane and toluene. The reaction time in this step is generally 5 minutes to 48 hours, preferably 1 hour to 24 hours. The reaction temperature in this step is generally -78°C to 100°C, preferably -78°C to 40°C.

[0535] (Step 6-2: Preparation step of compound (1-1c))

[0536] According to the method shown in step 2-3, compound (1-1c) can be produced from compound (6-1).

[0537] Preparation method 7

[0538] Among the compounds of formula (1), the compound of the following formula (1d) can be produced, for example, by the following production method.

[0539]

[0540] Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , m, n, r, s and Hy are as defined in item 1.

[0541] (Step 7-1: Preparation step of compound (1d))

[0542] Compound (1d) can be prepared by reacting compound (7-1) in the presence of triphosgene and a base in a suitable inert solvent, and then reacting the product with compound (1-1). The base used in this step can be appropriately selected from the bases exemplified below, including, for example, pyridine and triethylamine. The solvent used in this step can be appropriately selected from the solvents exemplified below, including, for example, dichloromethane and chloroform. The reaction time in this step is generally 5 minutes to 48 hours, preferably 30 minutes to 24 hours. The reaction temperature in this step is generally -78°C to 100°C, preferably 0°C to 80°C.

[0543] Preparation method 8

[0544] Among the compounds of formula (1), the compound of the following formula (1e) can be produced, for example, by the following production method.

[0545]

[0546] Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , m, n, r, s and Hy are as defined in item 1; and R 8 is C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkyl or cyclopropyl.

[0547] (Step 8-1: Preparation step of compound (1e))

[0548] According to the method shown in step 7-1, compound (1e) can be produced by reacting compound (8-1) with compound (1-1).

[0549] Preparation method 9

[0550] Among the compounds of formula (1), compounds of the following formula (1e) and formula (1f) can be produced, for example, by the following production method.

[0551]

[0552] Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , m, n, r, s and Hy are as defined in item 1; R 8 is C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or cyclopropyl; and Pro is an amino protecting group.

[0553] (Step 9-1: Preparation step of compound (9-2))

[0554] According to the method shown in step 7-1, compound (9-2) can be prepared by reacting compound (9-1) with compound (1-1). The amino protecting group Pro includes, for example, tert-butoxycarbonyl and benzyloxycarbonyl.

[0555] (Step 9-2: Preparation step of compound (1f))

[0556] According to the method shown in step 2-3, compound (1f) can be produced from compound (9-2).

[0557] (Step 9-3: Preparation step of compound (1e))

[0558] According to the method shown in Step 2-4, compound (1e) can be produced from compound (1f).

[0559] Among the starting materials and intermediates in each of the above-mentioned production methods, compounds not described in each method are commercially available, or can be produced by a skilled person in a known manner or a manner analogous thereto using commercially available materials.

[0560] The base used in each step of the above method should be appropriately selected based on the reaction, starting compound, etc., and includes, for example, alkaline bicarbonates such as sodium bicarbonate and potassium bicarbonate; alkaline carbonates such as sodium carbonate and potassium carbonate; metal hydrides such as sodium hydride and potassium hydride; alkali metal hydroxides such as sodium hydroxide and potassium hydroxide; alkali metal alkoxides such as sodium methoxide and sodium tert-butoxide; organic metal bases such as butyl lithium and lithium diisopropylamide; and organic bases such as triethylamine, diisopropylethylamine, pyridine, 4-dimethylaminopyridine (DMAP) and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU).

[0561] The solvent used in each step of the above method should be appropriately selected based on the reaction, starting compound, etc., including, for example, alcohol solvents such as methanol, ethanol and isopropanol; ketone solvents such as acetone and methyl ketone; halogenated hydrocarbon solvents such as dichloromethane and chloroform; ether solvents such as tetrahydrofuran (THF) and dioxane; aromatic hydrocarbon solvents such as toluene and benzene; aliphatic hydrocarbon solvents such as hexane and heptane; ester solvents such as ethyl acetate and propyl acetate; amide solvents such as N,N-dimethylformamide (DMF) and N-methyl-2-pyrrolidone (NMP); sulfoxide solvents such as dimethyl sulfoxide (DMSO); and nitrile solvents such as acetonitrile. The solvent used herein may be one of these solvents or a mixture of two or more solvents selected from these solvents. Moreover, if possible, in the reaction, an organic base such as diazabicycloundecene (DBU) may be used as the solvent used herein.

[0562] The compound of formula (1) and its intermediates can be separated and purified in a manner known to those skilled in the art, for example, including extraction, partitioning, reprecipitation, column chromatography (eg, silica gel column chromatography, ion exchange column chromatography and preparative liquid chromatography) and recrystallization.

[0563] The recrystallization solvent used herein includes, for example, alcohol solvents such as methanol, ethanol and 2-propanol; ether solvents such as ether; ester solvents such as ethyl acetate; aromatic hydrocarbon solvents such as benzene and toluene; ketone solvents such as acetone; halogenated solvents such as dichloromethane and chloroform; hydrocarbon solvents such as hexane; aprotic solvents such as dimethylformamide and acetonitrile; water; and mixed solvents thereof. As other purification methods, for example, the method described in Volume 1 of Series of Experimental Chemistry (Jikken Kagaku Kouza, edited by Chemical Society of Japan, Maruzen) can be used. Moreover, considering the structure of each starting compound, the structure determination of the present compound can be easily carried out by spectral analysis methods, such as nuclear magnetic resonance, infrared absorption technology and circular dichroism spectroscopy and / or mass spectrometry.

[0564] In addition, by appropriately modifying its functional groups, especially extending various side chains from amino, hydroxyl, carbonyl, halogen, etc.; and optionally carrying out the above protection and deprotection if necessary, each intermediate or each final product in the above preparation method can also be converted into another compound of the present invention. Modification of functional groups and extension of side chains can be carried out by conventional methods (for example, see Comprehensive Organic Transformations, RCLarock, John Wiley & Sons Inc. (1999) etc.).

[0565] The compound of the present formula (1) is sometimes an asymmetric compound, or sometimes has a substituent containing an asymmetric carbon. In this case, the compound has optical isomers. The present compound includes a mixture of these isomers and separated isomers, which can be prepared in a conventional manner. For example, by using a starting material with an asymmetric center or by introducing an asymmetric structure anywhere in the method, a compound with an asymmetric structure can be prepared. For example, in the case of optical isomers, optical isomers can be obtained by using optically active starting materials or by resolving an optical isomer mixture in an appropriate step. In the case where the compound of formula (1) or its intermediate has a basic functional group, its optical resolution includes, for example, a diastereomeric method, in which the compound is converted into a salt thereof by reacting with an optically active acid (e.g., monocarboxylic acid such as mandelic acid, N-benzyloxyalanine and lactic acid; dicarboxylic acid such as tartaric acid, o-diisopropyl-tartaric acid and malic acid; or sulfonic acid such as camphorsulfonic acid and bromocamphorsulfonic acid) in an inert solvent (e.g., alcohol such as methanol, ethanol and 2-propanol; ether solvent such as diethyl ether; ester solvent such as ethyl acetate; hydrocarbon solvent such as toluene; aprotic solvent such as acetonitrile; or a mixed solvent thereof). In the case where the compound of formula (1) or its intermediate has an acidic functional group such as a carboxyl group, the compound can also be optically resolved after forming its salt with an optically active amine (e.g., organic amine such as 1-phenylethylamine, bradykinin, quinidine, cinchonidine, cinchonine and strychnine).

[0566] The temperature for forming the salt is selected from the range of generally -50°C to the boiling point of the solvent used herein, preferably 0°C to the boiling point, and more preferably room temperature to the boiling point. In order to improve the optical purity, it is desirable to raise the temperature to near the boiling point of the solvent used herein. When collecting the precipitated crystals on the filter, optional cooling can increase the yield. The amount of the optically active acid or amine used herein is appropriately about 0.5-about 2.0 equivalents relative to the amount of the substance compound, preferably about 1 equivalent. If appropriate, the obtained crystals can be recrystallized in an inert solvent (e.g., an alcohol solvent such as methanol, ethanol and 2-propanol; an ether solvent such as diethyl ether; an ester solvent such as ethyl acetate; a hydrocarbon solvent such as toluene; an aprotic solvent such as acetonitrile; or a mixed solvent thereof) to obtain a highly pure salt thereof. Moreover, if appropriate, the optically resolved salt can also be treated with an acid or a base to obtain its free form.

[0567] In Lewy body diseases such as Parkinson's disease, abnormally aggregated α-synuclein is found in the brains of patients. Therefore, the drug of the present invention that inhibits or reduces the accumulation of α-synuclein aggregates is expected to play a role in improving these disease conditions.

[0568] In addition, it is believed that aggregates exhibit neurotoxicity, induce neuronal vulnerability and neuronal cell death, and cause the occurrence and development of pathology. Therefore, the drug of the present invention that inhibits neurotoxicity and neuronal cell death associated with α-synuclein aggregates is expected to play an effect of improving the pathology of Lewy body diseases such as Parkinson's disease.

[0569] The production of neurotransmitters is one of the functions of neurons, and a decrease in neurotransmitters indicates neuronal vulnerability. For example, in dopaminergic neurons, a decrease in the amount of tyrosine hydroxylase, which is involved in dopamine metabolism, indicates neuronal vulnerability.

[0570] In addition, EEG abnormalities have been reported in Lewy body diseases such as Parkinson's disease. The EEG is a manifestation of neural synchronization. Therefore, the drug of the present invention that normalizes the neuronal synchronization associated with α-synuclein aggregates is expected to play a role in improving the pathology of these diseases.

[0571] Neurospheroids for measuring the amount of α-synuclein aggregates can be prepared, for example, by three-dimensionally culturing neural stem cells prepared from human iPS cells or dopaminergic (DA) neural progenitor cells having a gene mutation associated with synucleinopathy under induction of neural differentiation. The amount of α-synuclein aggregates can be assessed by measuring the amount of α-synuclein with a high molecular weight by performing protein analysis using neurospheroids prepared by three-dimensional culture and α-synuclein antibodies.

[0572] Synchronous neural firing can be assessed using imaging analysis using fluorescent calcium probes in neurospheroids prepared by three-dimensional culture.

[0573] In addition, Parkinson's disease pathology can be reproduced by using the method of measuring the amount of α-synuclein aggregates and the method of measuring synchronized neural firing in neurospheroids, and drug evaluation for the effect of inhibiting or reducing the accumulation of α-synuclein aggregates in Parkinson's disease pathology can be performed.

[0574] Differentiation from human iPS cells with gene mutations associated with synucleinopathy into neural stem cells can be induced, for example, by culturing PLA2G6 gene mutant cells (clone name: 201B7, obtained from iPS Cell Research and Application, Kyoto University) established from an iPS cell line derived from a healthy human in StemFitAK03N medium (Ajinomoto Co., Inc., Basic03) at 37° C. and 5% CO 2 , and inducing the cultured product with PSC neuron induction medium (Thermo Fisher Scientific Inc., cat# A1647801).

[0575] For example, as a culture medium for neural stem cells, the following composition can be used.

[0576] <Medium composition for neural stem cells>

[0577] Neurobasal medium (Thermo Fisher Scientific Inc., 2113049)

[0578] Advanced DMEM / F-12 medium (Thermo Fisher Scientific Inc., 12634028)

[0579] Neural induction supplement (Thermo Fisher Scientific Inc., A1647801)

[0580] For example, differentiation from neural stem cells into neural spheroids can be induced by seeding neural stem cells (10,000 cells / well) in a 96-well round-bottom plate (Thermo Fisher Scientific Inc., cat#174929), culturing the cells in culture medium at 37°C and 5% CO2, and replacing half of the culture medium on the 2nd and 4th days after induction of differentiation.

[0581] For example, as a culture medium for neural stem cell neurospheres, the following composition can be used.

[0582] <Medium composition for neurospheres>

[0583] BrainPhys Neuronal Culture Medium (STEMCELL Technologies, cat#ST-05793)

[0584] NeuroCult SM1 Neuronal Supplement (STEMCELL Technologies, cat#05711)

[0585] N2 Supplement Supplement-A (STEMCELL Technologies, cat#07152)

[0586] 20ng / mL BDNF (PeproTech, Inc., cat#450-02)

[0587] 20ng / mL GDNF (PeproTech, Inc., cat#450-10)

[0588] 1mM dibutyryl cAMP (Nacalai Tesque, Inc., cat#11540-74)

[0589] 200 nM ascorbic acid (Nacalai Tesque, Inc., cat#03420-52)

[0590] Induced differentiation into dopaminergic progenitor cells from human iPS cells with a gene mutation associated with synucleinopathy can be performed, for example, by inducing dopaminergic progenitor cells from GBA1 gene homozygous mutation cells established from PLA2G6 gene mutation cells or iPS cell lines derived from healthy humans using a Dopaminergic Neuron Differentiation Kit (Thermo Fisher Scientific Inc., cat#A3147701).

[0591] For example, differentiation from dopaminergic progenitor cells into neurospheres can be induced by culturing cryopreserved dopaminergic progenitor cells at 37°C and 5% CO2 using a Floor Plate Cell Expansion Kit (Thermo Fisher Scientific Inc., cat#A3165801), seeding dopaminergic progenitor cells (10,000 cells / well) in a 96-well round-bottom plate (Thermo Fisher Scientific Inc., cat#174929), culturing the cells in culture medium at 37°C and 5% CO2, and replacing half of the culture every 3 or 4 days after induction of differentiation.

[0592] For example, as a culture medium for dopamine neurospheres derived from dopaminergic progenitor cells, the following composition can be used.

[0593] <Dopamine Neurospheroid Culture Medium Composition>

[0594] BrainPhys Neuronal Culture Medium (STEMCELL Technologies, cat#ST-05793)

[0595] Dopaminergic Neuron Maturation Supplement (Thermo Fisher Scientific Inc., cat# A3147401)

[0596] 20ng / mL BDNF (PeproTech, Inc., cat#450-02)

[0597] 20ng / mL GDNF (PeproTech, Inc., cat#450-10)

[0598] 1mM dibutyryl cAMP (Nacalai Tesque, Inc., cat#11540-74)

[0599] 200 nM ascorbic acid (Nacalai Tesque, Inc., cat#03420-52)

[0600] Measuring the amount of α-synuclein aggregates in neurospheroids can be performed, for example, by:

[0601] Remove the differentiated neurospheres from the culture medium.

[0602] A TBS solution (Nacalai Tesque, Inc., cat#12748-31) containing 1% TritionX-100 (Nacalai Tesque, Inc., cat#12967-32) was added thereto.

[0603] Extract the protein from the mixture using a sonicator, and

[0604] The extracted proteins were subjected to protein analysis (ProteinSimple, Inc., cat# SM-W008) using a Simple Western system under non-reducing conditions using an α-synuclein antibody (Thermo Fisher Scientific Inc., cat# AHB0261) to evaluate the quantification of the waveform shown at a molecular weight of approximately 300 kD.

[0605] Measurement of neural vulnerability in neurospheroids can be performed, e.g.

[0606] The differentiated dopamine neurospheres were transferred into TBS solution (Nacalai Tesque, Inc., cat#12748-31) containing 1% TritionX-100 (Nacalai Tesque, Inc., cat#12967-32).

[0607] Extract the protein from the mixture using a sonicator, and

[0608] The extracted proteins were subjected to protein analysis using a Simple Western system (Protein Simple, Inc., cat# SM-W004) using a tyrosine hydroxylase antibody (Millipore, cat# AB152) to evaluate the quantification of the waveform shown at a molecular weight of approximately 60 kD.

[0609] Measurement of neuronal cell death in neurospheroids can be performed, for example, by:

[0610] The differentiated dopamine neurospheres were transferred into TBS solution (Nacalai Tesque, Inc., cat#12748-31) containing 1% TritionX-100 (Nacalai Tesque, Inc., cat#12967-32).

[0611] Extract the protein from the mixture using a sonicator, and

[0612] The extracted proteins were subjected to protein analysis using a cleaved caspase 3 antibody (Cell Signaling Technology, Inc., cat#9664) with a Simple Western system (Protein Simple, Inc., cat#SM-W004) under reducing conditions to assess the quantification of the waveform shown at a molecular weight of approximately 20 kD.

[0613] Aberrant neural activity in neural spheroids can be measured, for example, by imaging neural spheroids in three-dimensional culture using fluorescent calcium probes to measure synchronized neural firing.

[0614] Synchronous neural firing in neural spheroids can be measured by, for example, imaging analysis using a measurement medium containing a fluorescent calcium probe (Molecular Devices, product name: FLIPR Calcium 6 Assay Batch Kit, cat# R8191).

[0615] The measurement medium used herein includes, for example, 20 mM Hepes (Thermo Fisher Scientific Inc., cat#15630-080) and Hank's buffer solution (Thermo Fisher Scientific Inc., cat#14065-056) containing 0.1% bovine serum albumin (Sigma-Aldrich, cat#A9576).

[0616] The compound of the present invention is useful as a drug for treating and / or preventing central nervous system diseases whose causes are associated with abnormal aggregation of proteins in the brain.

[0617] Central nervous system diseases whose causes are associated with abnormal aggregation of proteins in the brain include central nervous system diseases associated with tau, α-synuclein, TDP-43 or polyglutamine.

[0618] CNS diseases associated with tau include Alzheimer's disease and frontotemporal lobar degeneration; CNS diseases associated with α-synuclein aggregates include Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Gaucher disease, and infantile axonal dystrophy; CNS diseases associated with TDP-43 include amyotrophic lateral sclerosis and frontotemporal lobar degeneration; and CNS diseases associated with polyglutamine include Huntington's disease and spinocerebellar ataxia.

[0619] The compound of the present invention is preferably useful as a drug for the treatment and / or prevention of Alzheimer's disease, frontotemporal lobar degeneration, Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Gaucher disease, infantile axonal dystrophy, amyotrophic lateral sclerosis, Huntington's disease or spinocerebellar ataxia.

[0620] The compound of the present invention is more preferably useful as a drug for treating and or preventing diseases associated with α-synuclein aggregates.

[0621] Even more preferably, the compounds of the present invention can be used as drugs for the treatment and / or prevention of Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Gaucher disease or infantile neuroaxonal dystrophy.

[0622] In the present invention, "prevention" means administering the active ingredient of the present invention to a healthy subject who does not suffer from a disease, for example, the purpose of the administration is to prevent the occurrence of the disease. "Treatment" means administering the active ingredient of the present invention to a subject (i.e., a patient) diagnosed with the disease by a physician.

[0623] The compounds of the present invention and the drugs containing the compounds can be directly administered orally or parenterally, or can be administered as suitable pharmaceutical preparations. The preparation types include, for example, tablets, capsules, powders, granules, liquids, suspensions, injections, patches, poultices, etc., but are not limited thereto. The pharmaceutical preparations are prepared by conventional methods using pharmaceutically acceptable additives.

[0624] As additives, excipients, disintegrants, binders, fluidizers, lubricants, coating agents, solubilizers, solubilizing aids, thickeners, dispersants, stabilizers, sweeteners, flavors, etc. can be used depending on the purpose. Additives used herein include, for example, lactose, mannitol, crystalline cellulose, low-substituted hydroxypropyl cellulose, corn starch, partially pregelatinized starch, carmellose calcium, crosslinked carboxymethyl cellulose sodium, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinyl alcohol, magnesium stearate, sodium stearyl fumarate, polyethylene glycol, propylene glycol, titanium oxide, talc, etc.

[0625] The administration route that is the most effective for the treatment should be selected, which includes oral administration and parenteral administration, such as intravenous injection, wiping, inhalation and eye drops. Oral administration is preferred. The dosage form includes, for example, tablets and injections, and preferably tablets. The dosage of the pharmaceutical composition and the frequency of administration thereof may depend on the administration route, as well as the patient's disease, symptoms, age, body weight, etc., so it is impossible to simply define them. Usually, the present compound can be administered to adults at about 0.0001-about 5000 mg / day, preferably about 0.001-about 1000 mg / day, more preferably about 0.1-about 500 mg, and particularly preferably about 1-about 300 mg, which can be administered once a day or several times a day, preferably once to three times a day.

[0626] The compound of the present invention and the medicine containing it can be used together or in combination with different medicines to enhance the effect and / or reduce the side effect. They can be used together with the following medicines for the treatment of central nervous system diseases: such as L-dopa, dopamine agonists (e.g., ropinirole hydrochloride, apomorphine hydrochloride hydrate, etc.), MAO-B inhibitors (e.g., selegiline hydrochloride, etc.), catechol-O-methyltransferase (COMT) inhibitors (e.g., entacapone, etc.), α-Syn antibodies (e.g., Prasenimab, etc.) and pharmaceutically acceptable salts thereof. Hereinafter, the medicines that can be used together with the compounds of the present invention can be abbreviated as "different medicines (used together)".

[0627] The administration interval of the present compound, the drug containing it and the different drugs used together should not be limited. They can be given to the subject who needs it at the same time, or can be given at intervals. In addition, the present compound and the different drugs can be mixed as a combination drug. The dosage of the different drugs used together can be appropriately defined based on the dosage used clinically. The mixing ratio of the present compound and the different drugs used together can be appropriately defined depending on the subject to be treated, the route of administration and the disease, symptoms, combination, etc. of the subject. When the subject to be treated is human, different drugs used together can be used, for example, at a dosage of 0.01-100 parts by weight per portion of the present compound. In order to suppress its side effects, different drugs used together such as antiemetics, sleep-inducing drugs, and anticonvulsants can be used.

[0628] Example

[0629] The present invention is explained in more detail below by referring to reference examples, examples and tests; however, the technical scope of the present invention should not be limited thereto. In this specification, the term "example" or "reference example" sometimes means the compound itself, for example, "Example 1" means "the compound of Example 1", and "reference example 1" means "the compound of reference example 1". The names of compounds used in reference examples and examples should not always be based on the IUPAC naming system.

[0630] To simplify the description, the abbreviations shown below may sometimes be used in Reference Examples, Examples, and Tests.

[0631] Me:Methyl

[0632] Et: Ethyl

[0633] Pr:n-propyl

[0634] iPr: Isopropyl alcohol

[0635] DMF: N,N-dimethylformamide

[0636] THF: Tetrahydrofuran

[0637] TFA: trifluoroacetic acid

[0638] HATU: 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0639] The symbols used in NMR are defined as follows: s: singlet, d: doublet, dd: doublet of doublets, t: triplet, td: triplet of doublets, q: quartet, m: multiplet, br: broad, brs: broad singlet, brm: broad multiplet, J: coupling constant.

[0640] High performance liquid chromatography mass spectrometer; the measurement conditions of LCMS are as follows, where the observation results of mass spectrum [MS (m / z)] are shown as MH + , and the retention time is shown as Rt(min). In each measurement result, A or B is added as the analysis condition for measurement.

[0641] Analysis Condition A

[0642] Detection device: MS Detector: Waters ACQUITY SQ detector

[0643] HPLC: Waters ACQUITY UPLC

[0644] Column: ACQUITY UPLC BEH C18 1.7μm 2.1x30mm

[0645] Flow rate: 0.8 mL / min

[0646] Column temperature: 40°C

[0647] Wavelength: 254,220nm

[0648] Mobile Phase:

[0649] A: 0.06% formic acid / H2O

[0650] B: 0.06% formic acid / acetonitrile

[0651] Time program:

[0652] Step time (min)

[0653] 1 0.0-1.3 A:B=98:2-4:96

[0654] 2 1.3-1.5 A:B=4:96-98:2

[0655] Analysis Condition B

[0656] Detection device: Agilent 1200 Series, Agilent 6110 Quadrupole LCMS column: Xbridge C18 3.5 μm 4.6 x 50 mm

[0657] Flow rate: 1.8 mL / min

[0658] Wavelength: 254,214nm

[0659] Mobile Phase:

[0660] A: 10 mM ammonium bicarbonate aqueous solution

[0661] B: Acetonitrile

[0662] Time program:

[0663] Step time (min)

[0664] 1 0.0-1.5 A:B=90:10-5:95

[0665] Column temperature: 50°C

[0666] Reference Example 1

[0667] 5-(Piperidin-4-yl)-2-(trifluoromethyl)pyridine

[0668]

[0669] 5-bromo-2-(trifluoromethyl)pyridine (4.1 g), 1,1'-bis(diphenylphosphino)ferrocenepalladium chloride (0.66 g) and cesium carbonate (12 g) were added to a solution of 1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (6.7 g) in cyclopentyl methyl ether / water (4 / 1) (100 mL), and the mixture was stirred at 100° C. for 30 minutes. The reaction solution was cooled to room temperature, and then water was added to the reaction. The mixture was extracted with ethyl acetate. The organic layer was washed with brine and dried over magnesium sulfate. The solvent was removed from the dried organic layer in vacuo, and the residue was roughly purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate). The obtained crude product is dissolved in methanol (50mL), and 10% palladium carbon (55% wet, 0.30g) is added to the solution. The mixture is stirred at room temperature for 3 hours under a hydrogen atmosphere. The reaction solution is filtered based on diatomaceous earth and washed with methanol. The solvent in the filtrate is obtained by vacuum removal. The residue is dissolved in methanol (50mL) again, and 10% palladium carbon (55% wet, 0.30g) is added to the solution. The mixture is stirred at room temperature for 3 hours under a hydrogen atmosphere. The reaction solution is filtered based on diatomaceous earth and washed with methanol. The solvent in the filtrate is obtained by vacuum removal. The residue is dissolved in chloroform (10mL), and trifluoroacetic acid (20mL) is added to the solution. The mixture is stirred at room temperature for 1 minute, and the solvent and trifluoroacetic acid are removed from the solution in vacuum. The residue obtained is purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate -> ethyl acetate / methanol) to obtain Reference Example 1 (3.8g).

[0670] LC / MS ([M+H] + / Rt(min)):231.2 / 0.50(Analysis condition A)

[0671] Reference Example 2-11

[0672] According to the method described in Reference Example 1, the compounds shown in Table 1 were prepared from each corresponding starting compound.

[0673] [Table 1]

[0674]

[0675]

[0676] Reference Example 12

[0677] 5-(Azetidin-3-yl)-2-(trifluoromethyl)pyridine

[0678]

[0679] Zinc powder (0.33g) is added to a dry Schlenk tube, and the tube is purged with nitrogen. DMF (1.0mL) and iodine (51mg) are added to the tube under a nitrogen atmosphere, and the mixture is stirred at room temperature for 5 minutes. A solution of 3-iodoazetidine-1-carboxylic acid tert-butyl ester (0.19mL) in DMF (1.0mL) is added to the activated zinc solution, and the mixture is stirred at 40°C for 1 hour. A solution of 5-bromo-2-(trifluoromethyl) pyridine (0.23g) and tetrakis(triphenylphosphine) palladium (0) (58mg) in DMF (1.0mL) is added to the prepared alkyl zinc solution, and the mixture is stirred at 80°C for 2 hours. The reaction solution is cooled to 0°C, and the reaction is quenched by slowly dropping a saturated aqueous ammonium chloride solution to the reaction solution. The obtained precipitation is removed by suction filtration and washed with ethyl acetate. The filtrate is extracted with ethyl acetate, and the organic layer obtained is washed with brine and dried over magnesium sulfate. The solvent in the organic layer was removed in vacuo, and the crude product obtained was refluxed for 20 minutes with chloroform (1.0 mL) and trifluoroacetic acid (3.0 mL). The refluxed solution was cooled to room temperature, and the solvent and trifluoroacetic acid were removed in vacuo from the solution. The residue obtained was purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate) to obtain Reference Example 12 (75 mg).

[0680] LC / MS ([M+H] + / Rt(min)):203.1 / 0.47(Analysis condition A)

[0681] Reference Example 13

[0682] 4-[6-(Trifluoromethyl)pyridin-3-yl]piperidin-4-ol

[0683]

[0684] 5-bromo-2-(trifluoromethyl)pyridine (2.0g) is added to a dry two-necked flask, and the flask is purged with nitrogen. Dry THF (20mL) is added to the flask under a nitrogen atmosphere, and the mixture is cooled to -78°C. A solution of n-butyl lithium in hexane (2.6mol / L, 5.1mL) is added dropwise to the flask, and the mixture is stirred at -78°C for 1 hour. The prepared 1-Boc-4-piperidone (1.8g) solution in THF (10mL) is added dropwise to the reaction solution. After the addition is complete, the reaction solution is warmed to room temperature. After 1 hour, the reaction solution is cooled to 0°C, and the reaction is quenched by slowly dropping a saturated aqueous ammonium chloride solution to the reaction solution. The reaction solution is extracted with ethyl acetate, and the obtained organic layer is washed with brine and dried over magnesium sulfate. The solvent in the organic layer is removed in vacuo, and the residue obtained is roughly purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate 95:5-0:100). The obtained crude product was dissolved in chloroform (2.0 mL) and trifluoroacetic acid (4.0 mL), and the mixture was stirred at room temperature for 30 minutes. The solvent and trifluoroacetic acid were removed from the solution in vacuo, and the obtained residue was purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate -> ethyl acetate / methanol) to obtain Reference Example 13 (0.23 g).

[0685] LC / MS ([M+H] + / Rt(min)):247.1 / 0.46(Analysis condition A)

[0686] Reference Example 14

[0687] 5-(4-Fluoropiperidin-4-yl)-2-(trifluoromethyl)pyridine

[0688]

[0689] Reference Example 14

[0690] a) Preparation of tert-butyl 4-hydroxy-4-[6-(trifluoromethyl)pyridin-3-yl]piperidine-1-carboxylate (Compound W1)

[0691] 5-bromo-2-(trifluoromethyl)pyridine (2.0g) was added to a dry two-necked flask, and the flask was purged with nitrogen. Dry THF (20mL) was added to the flask under a nitrogen atmosphere, and the mixture was cooled to -78°C. A solution of n-butyl lithium in hexane (1.6mol / L, 6.8mL) was added dropwise to the flask, and the mixture was stirred at -78°C for 20 minutes. The prepared 1-Boc-4-piperidone (1.76g) solution in THF (20mL) was added dropwise to the reaction solution. After the addition was complete, the reaction solution was warmed to room temperature. After 1 hour, the reaction solution was cooled to 0°C, and the reaction was quenched by slowly dropping a saturated aqueous solution of ammonium chloride to the reaction solution. The reaction solution was extracted with ethyl acetate, and the obtained organic layer was washed with brine and dried over magnesium sulfate. The solvent in the organic layer was removed in vacuo, and the residue obtained was purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate) to obtain compound W1 (1.18g).

[0692] LC / MS ([M+H] + / Rt(min)):347.2 / 0.93(Analysis condition A)

[0693] b) Preparation of 5-(4-fluoropiperidin-4-yl)-2-(trifluoromethyl)pyridine (Reference Example 14)

[0694] Compound W1 (0.12g) was dissolved in chloroform (1.5mL), and bis(2-methoxyethyl)aminosulfur trifluoride (0.24mL) was added to the solution. The mixture was stirred at room temperature for 30 minutes. The reaction solution was concentrated in vacuo, and the residue obtained was roughly purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate). The obtained compound was dissolved in chloroform (0.50mL) and reacted with TFA (1.0mL). The reaction solution was stirred at room temperature for 5 minutes. The reaction solution was concentrated and azeotroped with toluene to remove residual TFA. The crude product obtained was purified by amino silica gel chromatography (eluting; hexane / ethyl acetate -> ethyl acetate / methanol) to obtain Reference Example 14 (18mg).

[0695] LC / MS ([M+H] + / Rt(min)):249.1 / 0.60(Analysis condition A)

[0696] Reference Examples 15-21

[0697] According to the method described in Reference Example 1, the compounds shown in Table 2 were prepared from each corresponding starting compound.

[0698] [Table 2]

[0699]

[0700] Example 1

[0701] (3-Methyloxetan-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone

[0702]

[0703] To a solution of Reference Example 1 (30 mg) in chloroform (1.0 mL) was added 3-methyloxetane-3-carboxylic acid (18 mg), triethylamine (27 μL) and HATU (60 mg), and the mixture was stirred at room temperature for 30 minutes. Cesium carbonate (42 mg) was added to the reaction solution, and the reaction solution was stirred again at room temperature for 30 minutes. The reaction solution was purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate -> ethyl acetate / methanol) to give Example 1 (41 mg).

[0704] LC / MS ([M+H] + / Rt(min)):329.2 / 0.75(Analysis condition A)

[0705] 1 H-NMR (400MHz, DMSO-d6) δ: 8.72 (1H, d, J = 1.6Hz), 8.02 (1H, dd, J = 2.0, 8.4Hz), 7.84 (1H, d, J = 8.8Hz), 4.83 (2H, dd, J = 6.4, 8.8Hz), 4.55 -4.52(1H,m),4.28(2H,t,6.8Hz),3.18-3.05(2H,m),3.01-2.93(1H,m),2.67(1H,t,J=12Hz),1.85-1.81(2H,brs),1.72-1.57(5H,m).

[0706] Alternatively, the compound of Example 1 can also be prepared according to the following method.

[0707] To a solution of the hydrochloride (100 mg) of Reference Example 1 in ethyl acetate (1.0 mL) were added 3-methyloxetane-3-carboxylic acid (52 mg), triethylamine (0.17 mL) and a solution of 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphaninane-2,4,6-trioxide in ethyl acetate (50%, 0.40 mL), and the mixture was stirred at room temperature for 1 hour. The reaction solution was purified by silica gel chromatography (eluting solvent; ethyl acetate / methanol) to obtain Example 1 (93 mg).

[0708] Example 2-32

[0709] According to the method described in Example 1, the compounds shown in Table 3 were prepared from each corresponding starting compound.

[0710] [Table 3]

[0711]

[0712]

[0713]

[0714]

[0715]

[0716]

[0717]

[0718]

[0719]

[0720]

[0721] Embodiment 33

[0722] (3-Methylazetidin-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone

[0723]

[0724] 1- (tert-butoxycarbonyl) -3- methylazetidine -3- carboxylic acid (56 mg), triethylamine (45 μ L), HATU (99 mg) and cesium carbonate (71 mg) are added to a solution of reference example 1 (50 mg) in chloroform (1.0 mL). The reaction solution is stirred at room temperature for 1 hour, and then roughly purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate -> ethyl acetate / methanol) to obtain a crude product. The obtained crude product is dissolved in chloroform (1.0 mL), and TFA (1.0 mL) is added thereto. The mixture is stirred at room temperature for 5 minutes. The reaction solution is concentrated and purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate -> ethyl acetate / methanol) to obtain Example 33 (65 mg).

[0725] LC / MS ([M+H] + / Rt(min)):328.2 / 0.60(Analysis condition A)

[0726] 1 H-NMR (400MHz, DMSO-d6) δ: 8.71 (1H, d, J = 2.0Hz), 8.00 (1H, dd, J = 8.4, 2.0Hz), 7.83 (1H, d, J = 8.4Hz), 4.53 (1H, d, J = 12.8Hz), 3.87 (2H, t, J =8.4Hz), 3.35-3.30(1H,m), 3.16-3.07(3H,m), 2.97(1H,tt,J=12,3.6Hz), 2.63(1H,t,J=12Hz), 1.84(2H,d,J=12.8Hz), 1.65-1.47(5H,m).

[0727] Alternatively, the compound of Example 33 can also be prepared according to the following method.

[0728] To a solution of the hydrochloride (100 mg) of Reference Example 1 in ethyl acetate (1.0 mL) was added 1-(tert-butoxycarbonyl)-3-methylazetidine-3-carboxylic acid (97 mg), triethylamine (0.17 mL) and a solution of 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphorinane-2,4,6-trioxide in ethyl acetate (50%, 0.40 mL), and the mixture was stirred at room temperature for 1 hour. The reaction solution was roughly purified by silica gel chromatography (eluting solvent; ethyl acetate / methanol) to obtain a crude product. The obtained crude product was dissolved in chloroform (1.0 mL), and TFA (1.0 mL) was added thereto. The mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated and purified by amino silica gel chromatography (eluting solvent; ethyl acetate / methanol) to obtain Example 33 (90 mg).

[0729] Examples 34-40

[0730] The compounds shown in Table 4 were prepared from each corresponding starting compound according to the method described in Example 33. In case the example compound was a salt, a salt-forming step was also included.

[0731] [Table 4]

[0732]

[0733]

[0734]

[0735] Embodiment 41

[0736] (1,3-Dimethylazetidin-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone hydrochloride

[0737]

[0738] To a solution of Example 33 (20 mg) in THF (0.50 mL) was added 36% aqueous formaldehyde solution (19 μL) and acetic acid (5.2 μL). Also, sodium triacetoxyborohydride (39 mg) was added to the solution while stirring the solution at room temperature. The reaction solution was stirred for 30 minutes and purified by amino silica gel chromatography (eluting solvent; hexane / ethyl acetate -> ethyl acetate / methanol). The obtained oily compound was dissolved in ethyl acetate (1 mL) and reacted with 4 mol / L hydrochloric acid in ethyl acetate at room temperature to give Example 41 (15 mg).

[0739] LC / MS ([M+H] + / Rt(min)):342.3 / 0.63(Analysis condition A)

[0740] 1 H-NMR (400MHz, DMSO-d6) δ: 10.6 (1H, brs), 8.72 (1H, d, J = 1.6Hz), 7.99 (1H, dd, J = 8.0, 1.6Hz), 7.86 (1H, d, J = 8.8Hz), 4.52 (1H, d, J = 12.8Hz), 4.13-4.1 2(2H,m),3.92-3.90(2H,m),3.32-3.29(1H,m),3.15(1H,t,J=12.8Hz),3.0 0(1H,tt,J=12,3.2Hz),2.72(3H,s),1.91-1.83(2H,m),1.68-1.50(5H,m).

[0741] Examples 42-48

[0742] The compounds shown in Table 5 were prepared from each corresponding starting compound according to the method described in Example 41. In the case where the example compound was not a salt, no salt-forming step was required.

[0743] [Table 5]

[0744]

[0745]

[0746] Embodiment 49

[0747] (Morpholin-4-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone

[0748]

[0749] To a solution of Reference Example 1 (30 mg) in chloroform (1.0 mL) were added morpholine-4-carbonyl chloride (18 μL) and N-ethyl-N-isopropylpropane-2-amine (34 μL), and the mixture was stirred at room temperature for 1 hour. The reaction solution was purified by amino silica gel chromatography (elution solvent; hexane / ethyl acetate -> ethyl acetate / methanol) to give Example 49 (44 mg).

[0750] LC / MS ([M+H] + / Rt(min)):344.2 / 0.79(Analysis condition A)

[0751] 1 H-NMR (400MHz, DMSO-d6) δ: 8.70 (1H, d, J = 2.0Hz), 7.98 (1H, dd, J = 8.0, 2.0Hz), 7.83 (1H, d, J = 8.0Hz), 3. 74(2H,d,J=13.6Hz),3.58(4H,m),3.15(4H,m),2.93-2.83(3H,m),1.82-1.79(2H,m),1.70-1.59(2H,m).

[0752] Examples 50-59

[0753] According to the method described in Example 1, the compounds shown in Table 6 were prepared from each corresponding starting compound.

[0754] [Table 6]

[0755]

[0756]

[0757]

[0758]

[0759] Embodiment 60

[0760] (1,4-oxazepan-4-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone

[0761]

[0762] To a solution of 1,4-oxazepane (61 mg) in dichloromethane (2.0 mL) was added pyridine (95 mg), and the mixture was cooled to 0° C. Triphosgene (72 mg) was added thereto, and the mixture was stirred at 0° C. for 30 minutes. Diisopropylethylamine (0.23 g) and Reference Example 1 (69 mg) were added to the reaction solution, and the reaction solution was stirred at room temperature for 1 hour. The reaction solution was concentrated and purified by preparative HPLC to give Example 60 (31 mg).

[0763] LC / MS ([M+H] + / Rt(min)):357.9 / 1.54(Analysis condition B)

[0764] 1 H-NMR (500MHz, CDCl3) δ: 8.63 (1H, s), 7.74-7.72 (1H, m), 7.66 (1H, d, J = 8.0Hz), 3.83-3.79 (6H, m), 3.55-3.5 2(4H,m),2.95-2.90(2H,m),2.86-2.81(1H,m),2.02-2.00(2H,m),1.91(2H,d,J=11.0Hz),1.80-1.72(2H,m).

[0765] Examples 61-63

[0766] According to the method described in Example 60, the compounds shown in Table 7 were prepared from each corresponding starting compound.

[0767] [Table 7]

[0768]

[0769]

[0770] Examples 64-65

[0771] The compounds shown in Table 8 were prepared from each corresponding starting compound according to the method described in Example 41. In the case where the example compound was not a salt, no salt-forming step was required.

[0772] [Table 8]

[0773]

[0774]

[0775] Embodiment 66

[0776] [3-Methyl-1-(2,2,2-trifluoroethyl)azetidin-3-yl]{4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone

[0777]

[0778] To a solution of Example 34 (46 mg) in THF (3.0 mL) were added 2,2,2-trifluoroethyl trifluoromethanesulfonate (42 mg) and triethylamine (71 mg), and the mixture was stirred at room temperature overnight. The reaction solution was concentrated and purified by preparative HPLC to give Example 66 (19 mg).

[0779] LC / MS ([M+H] + / Rt(min)):410.1 / 1.78(Analysis condition B)

[0780] 1 H-NMR (400MHz, CDCl3) δ: 8.59 (1H, d, J = 4.8Hz), 7.79 (1H, d, J = 8.0Hz), 7.51 ( 1H,dd,J=7.6,4.8Hz),4.79(1H,d,J=13.2Hz),3.54-3.50(4H,m),3.41(1H,d ,J=12.4Hz),3.26-3.17(2H,m),3.01(2H,q,J=9.2Hz),2.69(1H,t,J=12.8Hz ),1.90(2H,d,J=12.8Hz),1.81-1.77(1H,m),1.69(3H,s),1.67-1.57(1H,m).

[0781] Embodiment 67

[0782] (1-Cyclopropyl-3-methylazetidin-3-yl){4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone

[0783]

[0784] To a solution of Example 34 (46 mg) in ethanol (4.0 mL) were added (1-ethoxycyclopropyloxy)trimethylsilane (73 mg), acetic acid (13 mg) and sodium cyanoborohydride (44 mg), and the mixture was stirred overnight at 60° C. The reaction solution was cooled to room temperature and concentrated, and the residue was purified by preparative HPLC to give Example 67 (22 mg).

[0785] LC / MS ([M+H] + / Rt(min)):368.2 / 1.68(Analysis condition B)

[0786] 1 H-NMR (400MHz, CDCl3) δ: 8.58 (1H, dd, J = 4.4, 0.8Hz), 7.79 (1H, d, J = 8.0Hz), 7.50 (1H, dd, J = 8.0, 4.8Hz), 4.81 (1H, d, J = 12.8Hz), 3.53-3.50 (3H,m),3.38-3.31(2H,m),3.25-3.12(2H,m),2.76-2.65(1H,m),1.89-1.84(3H,m),1.71-1.61(2H,m),1.59(3H,s),0.38-0.35(4H,m).homo

[0787] Test 1: Neurospheres prepared by 3D culture of human iPS cells with PLA2G6 gene mutation Reproducibility test of Parkinson's disease pathology (accumulation of alpha-synuclein aggregates)

[0788] PLA2G6 gene mutant cells established from an iPS cell line derived from a healthy human (clone name: 201B7, obtained from iPS Cell Research and Application, Kyoto University) were cultured in StemFitAK03N medium (Ajinomoto Co., Inc., Basic03) at 37° C. and 5% CO 2 .

[0789] Neural stem cells were induced from iPS cells using PSC Neuron Induction Medium (Thermo Fisher Scientific Inc., cat#A1647801) to prepare cell stocks thereof.

[0790] Cryopreserved neural stem cells were cultured in a culture medium at 37° C. and 5% CO 2 . The culture medium used for neural stem cells herein had the following composition.

[0791] Culture medium composition for neural stem cells

[0792] Neurobasal medium (Thermo Fisher Scientific Inc., 2113049)

[0793] Advanced DMEM / F-12 medium (Thermo Fisher Scientific Inc., 12634028)

[0794] Neural induction supplement (Thermo Fisher Scientific Inc., A1647801)

[0795] Neural stem cells (10,000 cells / well) were seeded in 96-well round-bottom plates (Thermo Fisher Scientific Inc., cat#174929) and cultured in culture medium at 37°C and 5% CO2. Half of the culture medium was replaced every 3-4 days. The culture medium used for neurospheres herein had the following composition.

[0796] BrainPhys Neuronal Culture Medium (STEMCELL Technologies, cat#ST-05793)

[0797] NeuroCult SM1 Neuronal Supplement (STEMCELL Technologies, cat#05711)

[0798] N2 Supplement-A (STEMCELL Technologies, cat#07152)

[0799] 20ng / mL BDNF (PeproTech, Inc., cat#450-02)

[0800] 20ng / mL GDNF (PeproTech, Inc., cat#450-10)

[0801] 1mM dibutyryl cAMP (Nacalai Tesque, Inc., cat#11540-74)

[0802] 200 nM ascorbic acid (Nacalai Tesque, Inc., cat#03420-52)

[0803] The differentiated neurospheres were removed from the culture medium, a TBS solution (Nacalai Tesque, Inc., cat#12748-31) containing 1% TritionX-100 (Nacalai Tesque, Inc., cat#12967-32) was added thereto, and proteins were extracted from the mixture using a sonicator.

[0804] The extracted proteins were subjected to protein analysis (Protein Simple, Inc., cat# SM-W008) using a Simple Western system under non-reducing conditions using an α-synuclein antibody (Thermo Fisher Scientific Inc., cat# AHB0261) to measure the amount of α-synuclein aggregates and to assess the quantification of the waveform shown at a molecular weight of approximately 300 kD.

[0805] From the 7th day to the 9th day of culture, the amount of α-synuclein aggregates increased sharply. On the 9th day of culture, the amount of α-synuclein aggregates in the neurospheres derived from PLA2G6 mutant iPS cells was 5 times or more than that in the neurospheres derived from healthy iPS cells. After the 9th day of culture, a slow increase trend was shown. The results are shown in Figure 1 .

[0806] Test 2: Inhibition of α-synuclein with neurospheroids prepared from human iPS cells with a PLA2G6 gene mutation Assessment of protein aggregate accumulation

[0807] (1) Induction of differentiation into neural cells from human iPS cells

[0808] Neural stem cells were induced from PLA2G6 gene mutant iPS cells using PSC neuron induction medium (Thermo Fisher Scientific Inc., cat#A1647801). Neural spheroids were prepared from induced neural stem cells by a three-dimensional culture system and preserved with BrainPhys neuron medium (STEMCELL Technologies, cat#ST-05793) containing NeuroCult SM1 neuron supplement, N2 supplement-A, 20ng / mL BDNF, 20ng / mL GDNF, 1mM dibutyryl cAMP and 200nM ascorbic acid. Half of the culture medium was replaced on the 2nd and 4th day after induction of differentiation.

[0809] The test compound was diluted with the medium to twice the final concentration, and an equal volume of the twice concentrated solution was added to each well when half of the medium was replaced on day 4 after differentiation induction.

[0810] (2) Evaluation of the amount of α-synuclein aggregates

[0811] Proteins were extracted from neurospheres 9 days after induction of differentiation using a TBS solution containing 1% TritionX-100, and the extracted proteins were subjected to protein analysis using an α-synuclein antibody (Thermo Fisher Scientific Inc., cat#AHB0261) and a SimpleWestern system (Protein Simple, Inc., cat#SM-W008) to measure the amount of α-synuclein aggregates.

[0812] The amount of aggregates in the neurospheres added with each test compound was estimated assuming that the amount of aggregates in the neurospheres added with the DMSO solution was 100%. Table 9 shows the amount of aggregates of representative test compounds.

[0813] [Table 9]

[0814]

[0815]

[0816] Test 3: Reduction of α-synuclein using neurospheroids prepared from human iPS cells with a PLA2G6 gene mutation Assessment of protein aggregate accumulation

[0817] (1) Induction of differentiation into neural cells from human iPS cells

[0818] Neural stem cells were induced from PLA2G6 gene mutant iPS cells using PSC neuron induction medium (Thermo Fisher Scientific Inc., cat#A1647801). Neural spheroids were prepared from induced neural stem cells by a three-dimensional culture system and preserved with BrainPhys neuron medium (STEMCELL Technologies, cat#ST-05793) containing NeuroCult SM1 neuron supplement, N2 supplement-A, 20ng / mL BDNF, 20ng / mL GDNF, 1mM dibutyryl cAMP and 200nM ascorbic acid. Half of the culture medium was replaced every 3 or 4 days after differentiation was induced.

[0819] The test compound was diluted with the medium to twice the final concentration, and an equal volume of the twice concentrated solution was added to each well when half of the medium was replaced on day 10 after differentiation induction.

[0820] (2) Evaluation of the amount of α-synuclein aggregates

[0821] From the neurospheres 15 days after induction of differentiation, proteins were extracted with TBS solution containing 1% TritionX-100, and the extracted proteins were subjected to protein analysis using an α-synuclein antibody (Thermo Fisher Scientific Inc., cat#AHB0261) and a SimpleWestern system (Protein Simple, Inc., cat#SM-W008) to measure the amount of α-synuclein aggregates.

[0822] The amount of aggregates in the neurospheres added with each test compound was measured. The amount of aggregates in the test samples was estimated assuming that the amount of aggregates in the neurospheres added with the DMSO solution was 100%. Table 10 shows the amount of aggregates of representative test compounds.

[0823] [Table 10]

[0824]

[0825]

[0826] Test 4: Parkin in dopamine neurospheroids prepared from human iPS cells with a PLA2G6 gene mutation Reproducibility test of Mori's disease pathology (accumulation of α-synuclein aggregates)

[0827] Dopaminergic progenitor cells were induced from PLA2G6 gene mutant iPS cells using a Dopaminergic Neuron Differentiation Kit (Thermo Fisher Scientific Inc., cat#A3147701) to prepare a cell stock solution.

[0828] Cryopreserved dopaminergic progenitor cells were cultured using the Floor Plate Cell Expansion Kit (Thermo Fisher Scientific Inc., cat# A3165801) at 37°C and 5% CO2.

[0829] First, dopaminergic progenitor cells (10,000 cells / well) were seeded in a 96-well round-bottom plate (Thermo Fisher Scientific Inc., cat#174929) and cultured in a culture medium at 37°C and 5% CO2. Half of the culture medium was replaced every 3 or 4 days after differentiation induction. The culture medium used for dopamine neurospheres herein had the following composition.

[0830] BrainPhys Neuronal Culture Medium (STEMCELL Technologies, cat#ST-05793)

[0831] Dopaminergic Neuron Maturation Supplement (Thermo Fisher Scientific Inc., cat# A3147401)

[0832] 20ng / mL BDNF (PeproTech, Inc., cat#450-02)

[0833] 20ng / mL GDNF (PeproTech, Inc., cat#450-10)

[0834] 1mM dibutyryl cAMP (Nacalai Tesque, Inc., cat#11540-74)

[0835] 200 nM ascorbic acid (Nacalai Tesque, Inc., cat#03420-52)

[0836] The differentiated dopamine neurospheres were removed from the culture medium, a TBS solution (Nacalai Tesque, Inc., cat#12748-31) containing 1% TritionX-100 (Nacalai Tesque, Inc., cat#12967-32) was added thereto, and proteins were extracted from the mixture using a sonicator.

[0837] The extracted proteins were subjected to protein analysis (Protein Simple, Inc., cat# SM-W008) using a Simple Western system under non-reducing conditions using an α-synuclein antibody (Thermo Fisher Scientific Inc., cat# AHB0261) to measure the amount of α-synuclein aggregates and to evaluate the quantification of the waveform shown at a molecular weight of approximately 300 kD.

[0838] From the 10th day to the 21st day of culture, α-synuclein aggregates increased dramatically. On the 21st day of culture, the amount of α-synuclein aggregates in dopamine neurospheres derived from PLA2G6 mutant iPS cells was 5 times or more than that in dopamine neurospheres derived from healthy iPS cells. After the 21st day of culture, a slow increase trend was shown. The results are shown in Figure 2 .

[0839] Test 5: Inhibition of α- Assessment of synuclein aggregate accumulation.

[0840] (1) Induction of differentiation into neural cells from human iPS cells

[0841] Dopaminergic progenitor cells were induced from PLA2G6 gene mutant iPS cells using a dopaminergic neuron differentiation kit (Thermo Fisher Scientific Inc., cat#A3147701). Dopamine neurospheres were prepared from induced dopaminergic progenitor cells by a three-dimensional culture system and preserved with BrainPhys neuronal culture medium (STEMCELL Technologies, cat#ST-05793) containing NeuroCult SM1 neuronal supplement, N2 supplement-A, 20ng / mL BDNF, 20ng / mL GDNF, 1mM dibutyryl cAMP and 200nM ascorbic acid. Half of the culture medium was replaced every 3 or 4 days after differentiation was induced.

[0842] The test compound was diluted with the medium to twice the final concentration, and an equal volume of the twice concentrated solution was added to each well when half of the medium was replaced on day 21 after differentiation induction.

[0843] (2) Evaluation of the amount of α-synuclein aggregates

[0844] From dopamine neurospheres 26 days after induction of differentiation, proteins were extracted with TBS solution containing 1% TritionX-100, and the extracted proteins were subjected to protein analysis using an α-synuclein antibody (Thermo Fisher Scientific Inc., cat#AHB0261) and a Simple Western system (Protein Simple, Inc., cat#SM-W008) to measure the amount of α-synuclein aggregates.

[0845] The amount of aggregates in the neurospheres added with each test compound was measured. The amount of aggregates in the test samples was estimated assuming that the amount of aggregates in the neurospheres added with the DMSO solution was 100%. Table 11 shows the amount of aggregates (%) of representative test compounds.

[0846] [Table 11]

[0847]

[0848] Test 6: Neuronal expression of dopamine neurospheres prepared from human iPS cells with a PLA2G6 gene mutation Methods for reproducing vulnerabilities

[0849] (1) Induction of differentiation into neural cells from human iPS cells

[0850] Dopaminergic progenitor cells were induced from PLA2G6 gene mutant iPS cells using a dopaminergic neuron differentiation kit (Thermo Fisher Scientific Inc., cat#A3147701). Dopamine neurospheres were prepared from induced dopaminergic progenitor cells by a three-dimensional culture system and preserved with BrainPhys neuronal culture medium (STEMCELL Technologies, cat#ST-05793) containing NeuroCult SM1 neuronal supplement, N2 supplement-A, 20ng / mL BDNF, 20ng / mL GDNF, 1mM dibutyryl cAMP and 200nM ascorbic acid. Half of the culture medium was replaced every 3 or 4 days after induction of differentiation.

[0851] (2) Evaluation of the amount of tyrosine hydroxylase

[0852] Proteins were extracted from dopamine neurospheres 26 days after differentiation induction using a TBS solution containing 1% TritionX-100, and the extracted proteins were subjected to protein analysis using a tyrosine hydroxylase antibody (Millipore, cat#AB152) and a Simple Western system (Protein Simple, Inc., cat#SM-W008) to measure the amount of tyrosine hydroxylase. The results are shown in Figure 3 .

[0853] Test 7: Improving neuronal function using dopamine neurospheroids prepared from human iPS cells with a PLA2G6 gene mutation After vulnerability assessment

[0854] (1) Induction of differentiation into neural cells from human iPS cells

[0855] Dopaminergic progenitor cells were induced from PLA2G6 gene mutant iPS cells using a dopaminergic neuron differentiation kit (Thermo Fisher Scientific Inc., cat#A3147701). Dopamine neurospheres were prepared from induced dopaminergic progenitor cells by a three-dimensional culture system and preserved with BrainPhys neuronal culture medium (STEMCELL Technologies, cat#ST-05793) containing NeuroCult SM1 neuronal supplement, N2 supplement-A, 20ng / mL BDNF, 20ng / mL GDNF, 1mM dibutyryl cAMP and 200nM ascorbic acid. Half of the culture medium was replaced every 3 or 4 days.

[0856] The test compound was diluted with the medium to twice the final concentration, and an equal volume of the twice concentrated solution was added to each well when half of the medium was replaced on day 21 after differentiation induction.

[0857] (2) Evaluation of the amount of tyrosine hydroxylase

[0858] Proteins were extracted from dopamine neurospheres 26 days after differentiation induction using a TBS solution containing 1% TritionX-100, and the extracted proteins were subjected to protein analysis using a tyrosine hydroxylase antibody (Millipore, cat#AB152) and a Simple Western system (Protein Simple, Inc., cat#SM-W008) to measure the amount of tyrosine hydroxylase.

[0859] Test 8: Neuronal cells using neurospheres prepared from human iPS cells with PLA2G6 gene mutation Reproduction of death

[0860] (1) Induction of differentiation into neural cells from human iPS cells

[0861] Dopaminergic progenitor cells were induced from PLA2G6 gene mutant iPS cells using a dopaminergic neuron differentiation kit (Thermo Fisher Scientific Inc., cat#A3147701). Dopamine neurospheres were prepared from induced dopaminergic progenitor cells by a three-dimensional culture system and preserved with BrainPhys neuronal culture medium (STEMCELL Technologies, cat#ST-05793) containing NeuroCult SM1 neuronal supplement, N2 supplement-A, 20ng / mL BDNF, 20ng / mL GDNF, 1mM dibutyryl cAMP and 200nM ascorbic acid. Half of the culture medium was replaced every 3 or 4 days.

[0862] (2) Assessment of neuronal cell death

[0863] 10 μM dopamine was added to the culture medium 35 days after differentiation induction. Proteins were extracted from dopamine neurospheres 40 days after differentiation induction with a TBS solution containing 1% TritionX-100, and protein analysis was performed on the extracted proteins using a cleaved caspase 3 antibody (Cell Signaling Technology, Inc., cat#9664) using a Simple Western system (ProteinSimple, Inc., cat#SM-W008) to measure the amount of neuronal cell death. The results are shown in Figure 4 .

[0864] Test 9: Inhibition of neuronal proliferation using neurospheroids prepared from human iPS cells with a PLA2G6 gene mutation Assessment of cell death

[0865] (1) Induction of differentiation into neural cells from human iPS cells

[0866] Dopaminergic progenitor cells were induced from PLA2G6 gene mutant iPS cells using a dopaminergic neuron differentiation kit (Thermo Fisher Scientific Inc., cat#A3147701). Dopamine neurospheres were prepared from induced dopaminergic progenitor cells by a three-dimensional culture system and preserved with BrainPhys neuronal culture medium (STEMCELL Technologies, cat#ST-05793) containing NeuroCult SM1 neuronal supplement, N2 supplement-A, 20ng / mL BDNF, 20ng / mL GDNF, 1mM dibutyryl cAMP and 200nM ascorbic acid. Half of the culture medium was replaced every 3 or 4 days.

[0867] The test compound was diluted with the medium to twice the final concentration, and an equal volume of the twice concentrated solution having 10 μM dopamine was added to each well when half of the medium was replaced on day 35 after differentiation induction.

[0868] (2) Assessment of neuronal cell death

[0869] From dopamine neurospheres 40 days after induction of differentiation, proteins were extracted with TBS solution containing 1% TritionX-100, and the extracted proteins were subjected to protein analysis using a cleaved caspase 3 antibody (Cell Signaling Technology, Inc., cat#9664) with a Simple Western system (Protein Simple, Inc., cat#SM-W008) to measure the amount of neuronal cell death.

[0870] Test 10: Neurospheres prepared by three-dimensional culture of human iPS cells with GBA1 gene mutation Methods for reproducing Parkinson's disease pathology (accumulation of alpha-synuclein aggregates)

[0871] GBA1 gene homozygous mutant cells established from an iPS cell line derived from a healthy human were cultured in StemFitAK03N medium (Ajinomoto Co., Inc., Basic03) at 37° C. and 5% CO 2 .

[0872] Dopaminergic progenitor cells were induced from GBA1 homozygous mutant iPS cells using a Dopaminergic Neuron Differentiation Kit (Thermo Fisher Scientific Inc., cat#A3147701) to prepare cell stock solutions.

[0873] Cryopreserved dopaminergic progenitor cells were cultured in culture medium at 37° C. and 5% CO 2 using the Floor Plate Cell Expansion Kit (Thermo Fisher Scientific Inc., cat# A3165801).

[0874] Dopaminergic progenitor cells (10000 cells / well) were seeded in 96-well round-bottom plates (Thermo Fisher Scientific Inc., cat#174929) and cultured in culture medium at 37°C and 5% CO2. Half of the culture medium was replaced every 3-4 days after differentiation induction. The culture medium used for dopamine neurospheres herein had the following composition.

[0875] BrainPhys Neuronal Culture Medium (STEMCELL Technologies, cat#ST-05793)

[0876] Dopaminergic Neuron Maturation Supplement (Thermo Fisher Scientific Inc., cat# A3147401)

[0877] 20ng / mL BDNF (PeproTech, Inc., cat#450-02)

[0878] 20ng / mL GDNF (PeproTech, Inc., cat#450-10)

[0879] 1mM dibutyryl cAMP (Nacalai Tesque, Inc., cat#11540-74)

[0880] 200 nM ascorbic acid (Nacalai Tesque, Inc., cat#03420-52)

[0881] The differentiated DA neurospheres were removed from the culture medium, a TBS solution (Nacalai Tesque, Inc., cat#12748-31) containing 1% TritionX-100 (Nacalai Tesque, Inc., cat#12967-32) was added thereto, and proteins were extracted from the mixture using a sonicator.

[0882] α-synuclein antibody (Thermo Fisher Scientific Inc., cat#

[0883] AHB0261), and the extracted proteins were subjected to protein analysis using the Simple Western system under non-reducing conditions (Protein Simple, Inc., cat# SM-W008) to measure the amount of α-synuclein aggregates and to evaluate the quantification of the waveform shown at a molecular weight of approximately 300 kD.

[0884] From the 21st day to the 40th day of culture, the amount of α-synuclein aggregates increased dramatically. On the 40th day of culture, the amount of aggregates reached saturation, and no change in the amount was observed thereafter. The results are shown in Figure 5 .

[0885] Test 11: Reduction of α- Assessment of synuclein aggregate accumulation

[0886] (1) Induction of differentiation into neural cells from human iPS cells

[0887] Dopaminergic progenitor cells were induced from PLA2G6 gene mutant iPS cells using a dopaminergic neuron differentiation kit (Thermo Fisher Scientific Inc., cat#A3147701). Dopamine neurospheres were prepared from induced dopaminergic progenitor cells by a three-dimensional culture system and preserved with BrainPhys neuronal culture medium (STEMCELL Technologies, cat#ST-05793) containing NeuroCult SM1 neuronal supplement, N2 supplement-A, 20ng / mL BDNF, 20ng / mL GDNF, 1mM dibutyryl cAMP and 200nM ascorbic acid. Half of the culture medium was replaced every 3 or 4 days.

[0888] The test compound was diluted with the medium to twice the final concentration, and an equal volume of the twice concentrated solution was added to each well when half of the medium was replaced on day 40 after differentiation induction.

[0889] (2) Evaluation of the amount of α-synuclein aggregates

[0890] From dopamine neurospheres 44 days after induction of differentiation, proteins were extracted with TBS solution containing 1% TritionX-100, and the extracted proteins were subjected to protein analysis using an α-synuclein antibody (Thermo Fisher Scientific Inc., cat#AHB0261) and a Simple Western system (Protein Simple, Inc., cat#SM-W008) to measure the amount of α-synuclein aggregates.

[0891] The amount of aggregates in the neurospheres added with each test compound was measured. Assuming that the amount of aggregates in the neurospheres added with the DMSO solution was 100%, the amount of aggregates in the test samples was evaluated. Table 12 shows the amount of aggregates (%) of representative test compounds.

[0892] [Table 12]

[0893]

[0894] Test 12: Examination of neurospheres prepared by three-dimensional culture of human iPS cells with GBA1 gene mutation Check for abnormal synchronous discharge

[0895] GBA1 gene homozygous mutant cells established from an iPS cell line derived from a healthy human were cultured in StemFitAK03N medium (Ajinomoto Co., Inc., Basic03) at 37° C. and 5% CO 2 .

[0896] Dopaminergic progenitor cells were induced from GBA1 homozygous mutant iPS cells using a Dopaminergic Neuron Differentiation Kit (Thermo Fisher Scientific Inc., cat#A3147701) to prepare cell stock solutions.

[0897] Cryopreserved dopaminergic progenitor cells were cultured using the Floor Plate Cell Expansion Kit (Thermo Fisher Scientific Inc., cat# A3165801) at 37°C and 5% CO2.

[0898] Dopaminergic progenitor cells (10000 cells / well) were seeded in 96-well round-bottom plates (Thermo Fisher Scientific Inc., cat#174929) and cultured in culture medium at 37°C and 5% CO2. Half of the culture medium was replaced every 3-4 days after differentiation induction. The culture medium used for dopamine neurospheres herein had the following composition.

[0899] BrainPhys Neuronal Culture Medium (STEMCELL Technologies, cat#ST-05793)

[0900] Dopaminergic Neuron Maturation Supplement (Thermo Fisher Scientific Inc., cat# A3147401)

[0901] 20ng / mL BDNF (PeproTech, Inc., cat#450-02)

[0902] 20ng / mL GDNF (PeproTech, Inc., cat#450-10)

[0903] 1mM dibutyryl cAMP (Nacalai Tesque, Inc., cat#11540-74)

[0904] 200 nM ascorbic acid (Nacalai Tesque, Inc., cat#03420-52)

[0905] On or after the 40th day after differentiation induction, half of the culture medium was removed, and a measurement culture medium containing a fluorescent calcium probe (Molecular Devices, product name: FLIPR calcium 6 assay bulk kit, cat#R8191) equal to the remaining culture medium was added, and the mixture was left to stand for 30 minutes and then measured. The measurement culture medium used herein was 20mM Hepes (Thermo Fisher Scientific Inc., cat#15630-080) and Hank's buffer solution (Thermo Fisher Scientific Inc., cat#14065-056) containing 0.1% bovine serum albumin (Sigma-Aldrich, cat#A9576). One frame per second was taken.

[0906] Test 13: Improvement using dopamine neurospheroids prepared from human iPS cells with GBA1 gene mutation Evaluation of Synchronous Discharge Anomalies

[0907] (1) Induction of differentiation into neural cells from human iPS cells

[0908] Dopaminergic progenitor cells were induced from GBA1 gene mutant iPS cells using a dopaminergic neuron differentiation kit (Thermo Fisher Scientific Inc., cat#A3147701). Dopamine neurospheres were prepared from induced dopaminergic progenitor cells using a three-dimensional culture system and preserved in BrainPhys neuronal medium (STEMCELL Technologies, cat#ST-05793) containing NeuroCult SM1 neuronal supplement, N2 supplement-A, 20ng / mL BDNF, 20ng / mL GDNF, 1mM dibutyryl cAMP and 200nM ascorbic acid. Half of the culture medium was replaced every 3 or 4 days.

[0909] The test compound was diluted with the medium to twice the final concentration, and an equal volume of the twice concentrated solution was added to each well when half of the medium was replaced on day 40 after differentiation induction.

[0910] (2) Evaluation of synchronous discharge

[0911] On the 44th day after induction of differentiation, half of the dopamine neurosphere culture medium was removed, and a measurement culture medium containing a fluorescent calcium probe (Molecular Devices, product name: FLIPR calcium 6 assay bulk kit, cat#R8191) equal to the remaining culture medium was added, and the mixture was left to stand for 30 minutes and then measured. The measurement culture medium used herein was 20mM Hepes (Thermo Fisher Scientific Inc., cat#15630-080) and Hank's buffer solution (Thermo Fisher Scientific Inc., cat#14065-056) containing 0.1% bovine serum albumin (Sigma-Aldrich, cat#A9576). One frame per second was taken.

[0912] Industrial Applicability

[0913] The present compound has the effect of inhibiting or reducing the accumulation of α-synuclein aggregates, and thus the present compound can be used as a drug for treating or preventing central nervous system diseases, characterized by inhibiting or reducing the accumulation of abnormal protein aggregation in the brain. In addition, the present invention can be used as a method for reproducing Parkinson's disease pathology with neurospheroids, and a method for evaluating the amount of α-synuclein aggregates using a reproducing method.

[0914] As described above, the compound of formula (1) or a pharmaceutically acceptable salt thereof has the effect of inhibiting or reducing the aggregation of α-synuclein aggregates. Therefore, the compound of formula (1) or a pharmaceutically acceptable salt thereof can be used as a drug for treating or preventing central nervous system diseases associated with α-synuclein aggregates such as Parkinson's disease and Lewy body dementia.

Claims

1. A compound of formula (1) or a pharmaceutically acceptable salt thereof: in X is oxygen or NR 7 , R 7 is hydrogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkyl or cyclopropyl, Y is CH or nitrogen, m is 0, 1, or 2, n is 0 or 1, r is 0, 1, 2, 3, or 4, s is 0, 1, or 2, Provided that when s is 0, then Y is CH, and r is 1, 2, 3 or 4, When s is 1, then Y is CH, and r is 0, 1, 2 or 3, and When s is 2, then r is 1 or 2. R 1 is hydrogen, halogen, methyl or hydroxy, R 2 is hydrogen, halogen, methyl or hydroxy, R 3 is hydrogen or C 1-3 alkyl, R 4 is hydrogen or C 1-3 alkyl, Or, R 3 and R 4 may optionally together form a bridged methylene or a bridged ethylene, R 5 is halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and Hy is a pyridine ring, a pyridazine ring, a pyrimidine ring or a pyrazine ring, The premise is (I) When with R 5 and R 6 Hy is 5-fluoropyridin-2-yl, and m and n are 1, then r is 0, and s is 1, (II) When R 5 and R 6 When Hy is 6-methoxypyridin-3-yl, m and n are 1, and X is oxygen, then r is 0, and s is 1, (III) When R 5 and R 6 Hy is 5-methoxypyridin-2-yl, m and n are 1, and X is NR 7 When , r is 0, and s is 1, (IV) When R 5 is methyl, and R 6 When it is hydrogen, r is 0, and s is 1, but, (I') with R 5 and R 6 Hy is not 4,6-dimethylpyrimidin-2-yl or 5-bromopyrimidin-2-yl, (II') When with R 5 and R 6 When Hy is 5-chloropyridin-2-yl, m and n are 1, and R 1 and R 2 Not hydrogen, Provided that the following compounds are not included (III') (1-isopropylpiperidin-4-yl){3-(2-methoxypyridin-3-yl)pyrrolidin-1-yl}methanone, and (IV') (3-ethoxyoxetan-3-yl)[3-{4-(trifluoromethyl)pyrimidin-2-yl}pyrrolidin-1-yl]methanone.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein m is 1, and n is 1.

3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 are independently hydrogen, methyl or fluoro.

4. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 For hydrogen.

5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, wherein X is oxygen, NH or NMe.

6. The compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, wherein R 3 For hydrogen.

7. The compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein R 4 It is methyl or ethyl.

8. The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein Y is CH.

9. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein s is 1. 10 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein r is 0, and s is 1.

11. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is represented by formula (2): in X is oxygen, NH or NMe, R 4 is methyl or ethyl, R 5 is halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and Hy is a pyridine ring, a pyridazine ring, a pyrimidine ring or a pyrazine ring, The premise is that it has R 5 and R 6 The Hy of is not 5-chloropyridin-2-yl.

12. The compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof, wherein Hy is a pyridine ring.

13. The compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, wherein R 5 It is trifluoromethyl.

14. The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein Hy is pyridin-3-yl.

15. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein X is oxygen.

16. The compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, wherein R 4 It is methyl.

17. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein X is NH or NMe.

18. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein X is NMe.

19. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is selected from the following compounds: (3-methyloxetan-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, (3-methyloxetan-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, (3-methyloxetan-3-yl){4-[4-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, (3-methyloxetan-3-yl){4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, {4-[5-fluoro-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}(3-methyloxetan-3-yl)methanone, (3-methyloxetan-3-yl){4-[4-methyl-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, (1,3-dimethylazetidin-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, (1,3-dimethylazetidin-3-yl){4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, (1,3-dimethylazetidin-3-yl){4-[4-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, and (1,3-Dimethylazetidin-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone.

20. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is selected from the following compounds: (3-methyloxetan-3-yl){4-[6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, (3-methyloxetan-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, {4-[5-fluoro-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}(3-methyloxetan-3-yl)methanone, (3-methyloxetan-3-yl){4-[4-methyl-6-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, (1,3-dimethylazetidin-3-yl){4-[2-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone, and (1,3-Dimethylazetidin-3-yl){4-[5-(trifluoromethyl)pyridin-3-yl]piperidin-1-yl}methanone.

21. A medicament comprising the compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof as an active ingredient.

22. A drug for treating or preventing a central nervous system disease caused by abnormal protein aggregation in the brain, comprising as an active ingredient a compound of formula (1) or a pharmaceutically acceptable salt thereof: in X is oxygen or NR 7 , R 7 is hydrogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkyl or cyclopropyl, Y is CH or nitrogen, m is 0, 1, or 2, n is 0 or 1, r is 0, 1, 2, 3, or 4, s is 0, 1, or 2, Provided that when s is 0, then Y is CH, and r is 1, 2, 3 or 4, When s is 1, then Y is CH, and r is 0, 1, 2 or 3, and When s is 2, then r is 1 or 2. R 1 is hydrogen, halogen, methyl or hydroxy, R 2 is hydrogen, halogen, methyl or hydroxy, R 3 is hydrogen or C 1-3 alkyl, R 4 is hydrogen or C 1-3 alkyl, Or, R 3 and R 4 may optionally together form a bridged methylene or a bridged ethylene, R 5 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and Hy is a pyridine ring, a pyridazine ring, a pyrimidine ring or a pyrazine ring.

23. The medicament according to claim 22, wherein m is 1, and n is 1.

24. The drug according to claim 22 or 23, wherein R 1 and R 2 are independently hydrogen, methyl or fluoro.

25. The drug according to claim 22 or 23, wherein R 1 and R 2 For hydrogen.

26. The medicament according to any one of claims 22 to 25, wherein X is oxygen, NH or NMe.

27. The medicament according to any one of claims 22 to 26, wherein R 3 For hydrogen.

28. The medicament according to any one of claims 22 to 27, wherein R 4 It is methyl or ethyl.

29. The medicament according to any one of claims 22 to 28, wherein Y is CH.

30. The medicament according to any one of claims 22 to 29, wherein s is 1.

31. The medicament according to any one of claims 22 to 30, wherein r is 0, and s is 1.

32. The drug according to claim 22, wherein the compound is represented by formula (2): in X is oxygen, NH or NMe, R 4 is methyl or ethyl, R 5 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl, C 1-3 Alkoxy, R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl, C 1-3 Alkoxy, and Hy is a pyridine ring, a pyridazine ring, a pyrimidine ring or a pyrazine ring.

33. The medicament according to any one of claims 22 to 32, wherein Hy is a pyridine ring.

34. The medicament according to any one of claims 22 to 33, wherein R 5 It is trifluoromethyl.

35. The medicament according to any one of claims 22 to 34, wherein Hy is pyridin-3-yl.

36. The medicament according to any one of claims 22 to 35, wherein X is oxygen.

37. The medicament according to any one of claims 22 to 36, wherein R 4 It is methyl.

38. The medicament according to any one of claims 22 to 35, wherein X is NH or NMe.

39. The medicament according to any one of claims 22 to 35, wherein X is NMe.

40. The medicament according to any one of claims 22-39, wherein the central nervous system disease is associated with tau, α-synuclein, TDP-43 or polyglutamine.

41. The medicament according to any one of claims 22 to 39, wherein the central nervous system disease is Alzheimer's disease, frontotemporal lobar degeneration, Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Gaucher disease, infantile axonal dystrophy, amyotrophic lateral sclerosis, Huntington's disease, or spinocerebellar ataxia.

42. The medicament according to any one of claims 22-39, wherein the central nervous system disease is associated with α-synuclein.

43. The medicament according to any one of claims 22-39, wherein the central nervous system disease is associated with Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Gaucher disease or infantile neuroaxonal dystrophy.

44. A drug for treating or preventing a central nervous system disease whose cause is associated with abnormal aggregation of proteins in the brain, comprising a compound according to any one of claims 22 to 39 or a pharmaceutically acceptable salt thereof in combination with at least one agent selected from the group consisting of L-dopa, a dopamine agonist, a MAO-B inhibitor, a catechol-O-methyltransferase (COMT) inhibitor, an αSyn antibody and a pharmaceutically acceptable salt thereof.

45. A drug for treating or preventing a central nervous system disease whose cause is associated with abnormal aggregation of proteins in the brain, comprising a compound according to any one of claims 22 to 39 or a pharmaceutically acceptable salt thereof in combination with at least one agent selected from the group consisting of L-dopa, a dopamine agonist, a MAO-B inhibitor, a catechol-O-methyltransferase (COMT) inhibitor, an αSyn antibody and a pharmaceutically acceptable salt thereof.

46. ​​A method for preparing a compound of formula (3) or a pharmaceutically acceptable salt thereof: in R 4 is methyl or ethyl, R 5 is trifluoromethyl, R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and Hy is pyridin-3-yl, It includes the following steps 1; (Step 1) The compound of formula (4) or its salt: Where R 5 , R 6 and Hy are defined above, Condensation with a compound of formula (5) or a salt thereof: Where R 4 As defined above, and A is OH or halogen, To prepare the compound of formula (3) or a pharmaceutically acceptable salt thereof.

47. A method for preparing a compound of formula (6) or a pharmaceutically acceptable salt thereof: in R 4 Methyl, R 5 is trifluoromethyl, R 6 is hydrogen, halogen, C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 alkyl or C optionally substituted with 1 to 3 identical or different halogen atoms 1-3 Alkoxy, and Hy is pyridin-3-yl, It includes the following steps 1-3; (Step 1) The compound of formula (4) or its salt: Where R 5 , R 6 and Hy are defined above, Condensation with a compound of formula (7) or a salt thereof: Where R 4 As defined above, A is OH or halogen, and Pro is an amino protecting group, to prepare a compound of formula (8) or a pharmaceutically acceptable salt thereof: Where R 4 , R 5 , R 6 , Hy and Pro are as defined above, (Step 2) The amino protecting group in the compound of formula (8) or its salt is deprotected to prepare the compound of formula (9) or its salt: Where R 4 , R 5 , R 6 and Hy are defined above, (Step 3) The compound of formula (9) or a salt thereof is reacted with formaldehyde or an equivalent compound thereof in the presence of a reducing agent to prepare a compound of formula (6) or a pharmaceutically acceptable salt thereof.

Images