Huntington's disease treatment agents and therapeutic compositions

Abstract

The subject of this invention is to provide a Huntington's disease treatment agent or composition for Huntington's disease treatment, or a method of treating Huntington's disease, for which no treatment has been developed to date. The inventors of this invention have discovered that by differentiating striatal-type intermediate polyspinous neurons (MSNs) from iPS cells derived from Huntington's disease patients, and by administering a Huntington's disease treatment agent to the striatal-type MSNs exhibiting the pathological state of Huntington's disease, the pathological state of Huntington's disease in the striatal-type MSNs is improved. The Huntington's disease treatment agent comprises a compound of formula (1-1), a compound of formula (2-1), a compound of formula (3-1), a compound of formula (4-1), a pharmaceutically acceptable salt thereof, or a solvate thereof.

Description

Technical Field

[0001] The subject of this invention is to develop a therapeutic agent or composition for Huntington's disease. Background Technology

[0002] Huntington's disease is a hereditary neurodegenerative disease that follows an autosomal dominant inheritance pattern. It is characterized by involuntary movements such as choreiform movements, psychiatric symptoms, behavioral abnormalities, and cognitive impairment.

[0003] This disease is believed to be caused by the abnormal extension of the CAG repeat sequence in the Huntington's disease (HTT) gene (IT15 gene) located on chromosome 4. Because this genetic abnormality is hereditary and can be passed on to offspring, it is considered an autosomal dominant inheritance disorder. Pathological features of intracellular accumulation of abnormal HTT protein caused by this mutated HTT gene have been observed.

[0004] The onset and timing of Huntington's disease are thought to be related to the number of CAG repeats in the HTT gene. It is believed that Huntington's disease will develop when the number of CAG repeats is more than 40, and the more CAG repeats there are, the more likely it is to occur at a young age.

[0005] The incidence of the disease tends to vary slightly depending on race. It is reported that there are about 0.7 cases per 100,000 people in Japan and about 4 to 8 cases per 100,000 people in Caucasians. As for the specific number of patients, as of 2021, Japan reported 918 cases, while Europe and the United States are considered to have about 10 times that number.

[0006] Huntington's disease is believed to cause symptoms through the following mechanism: the abnormal HTT protein forms aggregates of various structural forms and exhibits different cytotoxicities, thereby causing degeneration of nerve cells in the striatum and cerebral cortex.

[0007] Its symptoms include initial symptoms, motor symptoms, involuntary movements, mental symptoms, and symptom progression. As each symptom...

[0008] • Initial symptoms: difficulty in performing fine motor skills, motor symptoms such as frowning or involuntary hand movements, irritability, or depression; and other mental symptoms / behavioral abnormalities.

[0009] • Motor symptoms: Initially, there are many cases where fine motor skills (such as writing) cannot be performed smoothly, and the same action cannot be continued. Symptoms such as dropping objects or falling occur. If the condition progresses, the gait becomes unstable, and symptoms such as tripping, falling, choking while eating, and difficulty speaking may occur.

[0010] • Involuntary movements: Rapid facial / limb movements, irregular involuntary hand movements, neck movements, frowning, tongue clicking, etc. (known as choreiform movements) that occur without one's own will;

[0011] • Mental symptoms: Unlike ordinary dementia, there is a tendency for impairment in the ability to plan and execute, and the ability to grasp the overall situation. Personality and behavioral changes may occur, such as becoming irritable or abnormally repeating the same behavior.

[0012] Currently, symptomatic treatments, such as medications to relieve symptoms, are used to address some symptoms of Huntington's disease, including choreiform movements, depressive symptoms, personality disorders, and irritability. Specifically, for choreiform movements, buphenazine, an antipsychotic drug that exhibits dopamine reuptake inhibition (release inhibition), is used.

[0013] However, there are currently no drugs available that aim to cure Huntington's disease (improvement in motor function, psychiatric symptoms, and cognitive function), and no cure-oriented treatment has yet been established. Therefore, there is a desire to develop drugs or treatments that can improve motor function, psychiatric symptoms, and cognitive function.

[0014] Existing technical documents

[0015] Non-patent literature

[0016] Non-patent literature 1: Wu M., et al., A Chemical Recipe for Generation of Clinical-Grade Striatal Neurons from hESCs., Stem cell reports, 11(3), 2018

[0017] Non-patent literature 2: Victor MB., et al., Generation of human striatal neurons by microRNA-dependent direct conversion of fibroblasts., Neuron, 84(2), 2015 Summary of the Invention

[0018] The problem the invention aims to solve

[0019] The subject of this invention is to provide a Huntington's disease treatment agent or composition for Huntington's disease treatment, or a method of treating Huntington's disease, for which no treatment has been developed to date.

[0020] Solution for solving the problem

[0021] The inventors of this invention discovered that by inducing the differentiation of mid-spiny neurons (MSNs) in the striatum from iPS cells derived from Huntington's disease patients, and by administering the following Huntington's disease therapeutic agent to the mid-spiny neurons (MSNs) in the striatum exhibiting the pathological state of Huntington's disease, the pathological state of the mid-spiny neurons (MSNs) in the striatum is improved.

[0022] Based on this insight, the present invention demonstrates that Huntington's disease can be treated by a Huntington's disease treatment agent comprising a compound of formula (1-1), a compound of formula (2-1), a compound of formula (3-1) or a compound of formula (4-1), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0023] More specifically, in order to solve the aforementioned problems, this application provides the following approach:

[0024] [1]: A Huntington's disease treatment comprising a compound represented by formula (1-1), a compound represented by formula (2-1), a compound represented by formula (3-1), a compound represented by formula (4-1), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0025]

[0026] In equation (1-1), R 1 Each of these can independently represent an alkyl group or a 4-hydroxyphenylethyl group having 1 to 6 carbon atoms, where n represents an integer from 1 to 3.

[0027]

[0028] In equation (2-1), R 21 For H or -CH3,

[0029] R 22 [Selected from the group consisting of H, alkyl groups having 1 to 6 carbon atoms, alkoxy groups having 1 to 6 carbon atoms, and halogens]

[0030]

[0031] In equation (3-1),

[0032] Y represents N or A 2 -C,

[0033] Z represents N or CH.

[0034] m can be 0, 1, 2, or 3.

[0035] B represents O, S, or NR. 33 ,

[0036] A 1 and A 2 Independently representing an alkyl group having 1 to 6 optional carbon atoms, an alkenyl group having 2 to 6 optional carbon atoms, an alkynyl group having 2 to 6 optional carbon atoms, a cycloalkyl group having 3 to 7 optional carbon atoms, an aryl group having 1 to 6 optional carbon atoms, a heterocycloalkyl group having 3 to 7 optional carbon atoms, a heteroaryl group having 1 to 6 optional carbon atoms, or a heteroaryl group having 1 to 6 optional carbon atoms, or H, halogen, cyano, trifluoromethyl, an alkoxy group having 1 to 6 carbon atoms, an alkylthio group having 1 to 6 carbon atoms, or -NR x R y ,

[0037] Among them, R x and R y Independently representing H, hydrocarbon group, or heterocyclic group, or R x and R y Together they represent alkylene groups having 2 to 6 carbon atoms.

[0038] R 31 -CH2CHR 34 NR 35 R 36 Or the following formula:

[0039]

[0040] R 32 R 33 R 34 R 35 R 36 and R 37 [Independently representing H or an alkyl group having 1 to 6 carbon atoms],

[0041]

[0042] In equation (4-1),

[0043] R 41 and R 42 Selected from alkyl groups having 1 to 6 carbon atoms, aryl groups having 6 to 10 carbon atoms, and alkyl groups having 1 to 6 carbon atoms substituted with aryl groups having 6 to 10 carbon atoms.

[0044] R 43 Selected from alkyl groups having 1 to 6 carbon atoms,

[0045] M is selected from halogens and alkyl sulfates.

[0046] [2]: According to the Huntington's disease treatment agent described in [1], n is 2 in the aforementioned formula (1-1).

[0047] [3]: The Huntington's disease treatment according to [1] or [2], wherein R1 in the aforementioned formula (1-1) is n-propyl.

[0048] [4]: The Huntington's disease treatment agent according to [1] or [2], wherein the compound represented by the aforementioned formula (1-1) is the compound represented by the following formula (1-2):

[0049]

[0050] (4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indol-2-one).

[0051] [5]: The Huntington's disease treatment agent according to claim [4], wherein the pharmaceutically acceptable salt of the compound represented by formula (1-1) is the hydrochloride salt of the compound represented by formula (1-2), namely 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one hydrochloride.

[0052] [6]: The Huntington's disease treatment agent according to [1] or [2], wherein the compound represented by the aforementioned formula (2-1) is the compound represented by the following formula (2-2):

[0053]

[0054] (2-[3-[4-(m-chlorophenyl)-1-piperazinyl]propyl]-s-triazolo[4,3-a]pyridine-3(2H)-one).

[0055] [7]: The Huntington's disease treatment agent according to [6], wherein the pharmaceutically acceptable salt of the compound shown in formula (2-1) is the hydrochloride salt of the compound shown in formula (2-2) (2-[3-[4-(m-chlorophenyl)-1-piperazinyl]propyl]-s-triazolo[4,3-a]pyridine-3(2H)-one hydrochloride).

[0056] [8]: The Huntington's disease treatment agent according to [1] or [2], wherein the compound represented by the aforementioned formula (3-1) is the compound represented by the following formula (3-2):

[0057]

[0058] (N,N-Dimethyl-2-[5-(1H-1,2,4-triazol-1-ylmethyl)-1H-indol-3-yl]ethylamine).

[0059] [9]: According to the Huntington's disease treatment agent described in [8], wherein the pharmaceutically acceptable salt of the compound shown in formula (3-1) is the benzoate of the compound shown in formula (3-2) (N,N-dimethyl-2-[5-(1H-1,2,4-triazol-1-ylmethyl)-1H-indol-3-yl]ethylamine benzoate).

[0060]

[10] : The Huntington's disease treatment agent according to [1] or [2], wherein the compound represented by the aforementioned formula (4-1) is the compound represented by the following formula (4-2):

[0061]

[0062] (3-Dimethylcarbamoyloxy-1-methylpyridinium bromide).

[0063]

[11] : A pharmaceutical composition for treating Huntington's disease, comprising, as an active ingredient, a compound of formula (1-1), a compound of formula (2-1), a compound of formula (3-1), a compound of formula (4-1), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0064]

[0065] [In formula (1-1), R1 independently represents an alkyl group or a 4-hydroxyphenylethyl group with 1 to 6 carbon atoms, and n represents an integer from 1 to 3.]

[0066]

[0067] In equation (2-1), R 21 For H or -CH3,

[0068] R 22 [Selected from the group consisting of H, alkyl groups having 1 to 6 carbon atoms, alkoxy groups having 1 to 6 carbon atoms, and halogens]

[0069]

[0070] In equation (3-1),

[0071] Y represents N or A 2 -C,

[0072] Z represents N or CH.

[0073] m can be 0, 1, 2, or 3.

[0074] B represents O, S, or NR. 33 ,

[0075] A 1 and A2 Independently representing an alkyl group having 1 to 6 optional carbon atoms, an alkenyl group having 2 to 6 optional carbon atoms, an alkynyl group having 2 to 6 optional carbon atoms, a cycloalkyl group having 3 to 7 optional carbon atoms, an aryl group having 1 to 6 optional carbon atoms, a heterocycloalkyl group having 3 to 7 optional carbon atoms, a heteroaryl group having 1 to 6 optional carbon atoms, or a heteroaryl group having 1 to 6 optional carbon atoms, or H, halogen, cyano, trifluoromethyl, an alkoxy group having 1 to 6 carbon atoms, an alkylthio group having 1 to 6 carbon atoms, or -NR x R y ,

[0076] Among them, R x and R y Independently representing H, hydrocarbon group, or heterocyclic group, or R x and R y Together they represent alkylene groups having 2 to 6 carbon atoms.

[0077] R 31 -CH2CHR 34 NR 35 R 36 Or the following formula:

[0078]

[0079] R 32 R 33 R 34 R 35 R 36 and R 37 [Independently representing H or an alkyl group having 1 to 6 carbon atoms],

[0080]

[0081] In equation (4-1),

[0082] R 41 and R 42 Selected from alkyl groups having 1 to 6 carbon atoms, aryl groups having 6 to 10 carbon atoms, and alkyl groups having 1 to 6 carbon atoms substituted with aryl groups having 6 to 10 carbon atoms.

[0083] R 43 Selected from alkyl groups having 1 to 6 carbon atoms,

[0084] M is selected from halogens and alkyl sulfates.

[0085] The effects of the invention

[0086] The Huntington's disease therapeutic agent of the present invention is a compound obtained by analyzing the striatal type of medium-sized polyspinous neurons (MSNs) induced by differentiation of iPS cells derived from Huntington's disease patients as a pathological state model and screening them with the characteristic phenotype of human pathological state as an evaluation item. Therefore, it can provide a therapeutic agent with high therapeutic effect on Huntington's disease. Attached Figure Description

[0087] Figure 1 This is a diagram illustrating a cell culture protocol used to create striatal-type medium-sized multispinous neurons (MSNs).

[0088] Figure 2 This is an image showing DARPP32 and DRD2 positive cells as MSN markers, obtained by immunostaining of cells derived from healthy individuals after differentiation induction.

[0089] Figure 3 This is a graph showing the results of immunostaining of differentiation-induced cells and the study of the percentage of DARPP32 and DRD2 positive cells as MSN markers.

[0090] Figure 4 This figure shows the results of an investigation into the phenotype of striatal MSN derived from disease-specific iPS cells in Huntington's disease, specifically LDH leakage. The figure demonstrates that the LDH leakage rate is significantly increased in MSN cells derived from disease-specific iPS cells.

[0091] Figure 5 This is a diagram illustrating the phenotype of striatal MSNs derived from disease-specific iPS cells in Huntington's disease. The diagram shows that neurite length is significantly reduced in MSN cells derived from disease-specific iPS cells.

[0092] Figure 6 This is a graph showing the results of a study on the phenotype of striatal MSNs derived from disease-specific iPS cells in Huntington's disease, specifically the neurite length. The graph shows that the neurite length in MSNs derived from disease-specific iPS cells significantly decreases over time.

[0093] Figure 7 This is a diagram illustrating the phenotype of striatal MSN derived from disease-specific iPS cells in Huntington's disease, showing significant accumulation of HTT aggregates in MSN cells derived from disease-specific iPS cells.

[0094] Figure 8The figure shows the results of an investigation into the accumulation of HTT aggregates, a phenotype of striatal MSN derived from disease-specific iPS cells in Huntington's disease. The figure shows that HTT aggregates accumulate significantly over time in MSN cells derived from disease-specific iPS cells.

[0095] Figure 9 This diagram illustrates the steps involved in screening compounds for the treatment of Huntington's disease using striatal-type MSNs derived from disease-specific iPS cells.

[0096] Figure 10-1 This is a graph showing the improvement rate (%) of pathological status calculated based on three indicators when eight compounds obtained through compound screening were applied to cells (HD1.CAG180) with a CAG replication number of 180.

[0097] Figure 10-2 This is a graph showing the improvement rate (%) of pathological status calculated based on three indicators when eight compounds obtained through compound screening were applied to cells (HD2.CAG50) with 50 repetitions of CAG.

[0098] Figure 11 This is a graph showing the effects of various concentrations of ropinirole on neurite retraction.

[0099] Figure 12 This is a graph showing the effects of various concentrations of ropinirole on HTT aggregates.

[0100] Figure 13 This is a diagram illustrating the steps involved in studying the effects of ropinirole using MSNs derived from striatal types of Huntington's disease-specific iPS cells.

[0101] Figure 14 This is a graph showing the effect of various concentrations of ropinirole on LDH leakage when using striatal-type MSNs derived from Huntington's disease-specific iPS cells.

[0102] Figure 15 This is a graph showing the effect of various concentrations of ropinirole on LDH leakage when using striatal-type MSNs derived from Huntington's disease-specific iPS cells.

[0103] Figure 16 This is a graph showing the effect of various concentrations of ropinirole on HTT aggregates when using striatal-type MSNs derived from Huntington's disease-specific iPS cells.

[0104] Figure 17 This is a graph showing the effect of various concentrations of ropinirole on HTT aggregates when using striatal-type MSNs derived from Huntington's disease-specific iPS cells.

[0105] Figure 18 This is a graph showing that ropinirole at various concentrations is more effective than bromocriptine at the same concentration range in reducing LDH leakage when using striatal MSNs derived from Huntington's disease-specific iPS cells.

[0106] Figure 19 This is a graph showing the effect of various concentrations of ropinirole on LDH leakage induced by the addition of glutamate (50 μM) when using striatal-type MSNs derived from Huntington's disease-specific iPS cells.

[0107] Figure 20 This is a graph showing the improvement rate of LDH leakage after administration of 300 nM ropinirole relative to the use of multiple Huntington's disease-specific iPS cell-derived striatal MSNs. Detailed Implementation

[0108] [Screening of Huntington's Disease Treatment Agents and Drug Compositions for Huntington's Disease Treatment]

[0109] In this invention, iPS cells derived from cells collected from Huntington's disease patients are prepared. These iPS cells are then differentiated to induce medium-sized polyspinous neurons (MSNs) in the striatum. Using MSNs that reflect the pathological state of Huntington's disease, compounds in a compound library are screened based on evaluation criteria such as improving neurite retraction, inhibiting HTT aggregate number, and exhibiting neuroprotective effects (increased neuronal survival). This yields compounds that can improve the pathological state of Huntington's disease in MSNs and have therapeutic effects on Huntington's disease.

[0110] Regarding the method for inducing differentiation of medium-sized polyspinous neurons (MSNs) in the striatum, iPS cells were induced to differentiate into lateral basal nucleus primordia through dual SMAD inhibition (e.g., LDN-193189 and SB431542) and Wnt inhibition (e.g., Noggin and XAV939) with the addition of Sonic Hedgehog factor. Brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), and ascorbic acid were then added to obtain MSNs (Non-Patent Literature 1). However, due to the long time required for differentiation induction and the low efficiency of differentiation induction, which contributes to intercellular differences, a gene expression-based differentiation induction method using fibroblasts was reported (Non-Patent Literature 2). However, since the method is based on fibroblasts, the inventors of this invention have disclosed that by using iPS cells to induce gene expression of various transcription factors, the differentiation of medium-sized multispinous neurons (MSNs) in the striatum can be induced in a short time in multiple cell lines.

[0111] The compound library used in the screening can be of any type, such as a compound library that has undergone clinical trials for various diseases and whose safety has been confirmed.

[0112] In this invention, a compound library was screened using this method, and ropinirole, trazodone, rizatriptan, and pyridostigmine were selected as compounds that exhibited effects such as improving neurite retraction, inhibiting HTT aggregate number, and neuroprotective effects (increasing neuronal survival). Based on this insight, the inventors of this invention completed the present invention.

[0113] [Huntington's disease treatment agents, Huntington's disease treatment drug compositions]

[0114] In this invention, for pharmaceutical use of the compounds represented by formula (1-1), the compounds represented by formula (2-1), the compounds represented by formula (3-1), the compounds represented by formula (4-1), their pharmaceutically acceptable salts, or their solvates, the present invention provides: a Huntington's disease treatment agent comprising the above-mentioned compounds, their pharmaceutically acceptable salts, or their solvates; or a Huntington's disease treatment pharmaceutical composition comprising the above-mentioned compounds, their pharmaceutically acceptable salts, or their solvates.

[0115]

[0116] [In formula (1-1), R1 independently represents an alkyl group or a 4-hydroxyphenylethyl group with 1 to 6 carbon atoms, and n represents an integer from 1 to 3.]

[0117]

[0118] In equation (2-1), R 21 For H or -CH3,

[0119] R 22 [Selected from the group consisting of H, alkyl groups having 1 to 6 carbon atoms, alkoxy groups having 1 to 6 carbon atoms, and halogens]

[0120]

[0121] In equation (3-1),

[0122] Y represents N or A 2 -C,

[0123] Z represents N or CH.

[0124] m can be 0, 1, 2, or 3.

[0125] B represents O, S, or NR. 33 ,

[0126] A 1 and A 2 Independently representing an alkyl group having 1 to 6 optional carbon atoms, an alkenyl group having 2 to 6 optional carbon atoms, an alkynyl group having 2 to 6 optional carbon atoms, a cycloalkyl group having 3 to 7 optional carbon atoms, an aryl group having 1 to 6 optional carbon atoms, a heterocycloalkyl group having 3 to 7 optional carbon atoms, a heteroaryl group having 1 to 6 optional carbon atoms, or a heteroaryl group having 1 to 6 optional carbon atoms, or H, halogen, cyano, trifluoromethyl, an alkoxy group having 1 to 6 carbon atoms, an alkylthio group having 1 to 6 carbon atoms, or -NR x R y ,

[0127] Among them, R x and R y Independently representing H, hydrocarbon group, or heterocyclic group, or R x and R y Together they represent alkylene groups having 2 to 6 carbon atoms.

[0128] R 31 -CH2CHR 34 NR 35 R36 Or the following formula:

[0129]

[0130] R 32 R 33 R 34 R 35 R 36 and R 37 [Independently representing H or an alkyl group having 1 to 6 carbon atoms],

[0131]

[0132] In equation (4-1),

[0133] R 41 and R 42 Selected from alkyl groups having 1 to 6 carbon atoms, aryl groups having 6 to 10 carbon atoms, and alkyl groups having 1 to 6 carbon atoms substituted with aryl groups having 6 to 10 carbon atoms.

[0134] R 43 Selected from alkyl groups having 1 to 6 carbon atoms,

[0135] M is selected from halogens and alkyl sulfates.

[0136] The diseases that can be treated with the Huntington's disease therapeutic agent or the Huntington's disease therapeutic composition of the present invention can be any of the following: motor function symptoms such as chorea and involuntary movements, cognitive function symptoms such as dementia, and psychiatric symptoms. Furthermore, based on the pathological characteristics of patients with Huntington's disease, even patients with various neurodegenerative diseases accompanied by the accumulation of HTT protein aggregates, or those with abnormal accumulation of other protein aggregates (e.g., β-amyloid, α-synuclein, 43kDa TAR DNA-binding protein, sarcoma fusion protein, etc.), can be treated. Moreover, based on the genetic characteristics of patients with Huntington's disease, even patients with any type of Huntington's disease, such as those with mutations in the HTT gene, or those with mutations in other genes reported to have aggregate protein accumulation (e.g., β-amyloid, α-synuclein, 43kDa TAR DNA-binding protein, sarcoma fusion protein, etc.), can be treated.

[0137] In the compounds of formula (1-1) above, which are the active ingredients of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention, n in formula (1-1) can be 1, 2, or 3. When the active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention is ropinirole as described below, n is 2 in formula (1-1). Therefore, the active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention can also be a compound in formula (1-1) where n is 2.

[0138] Furthermore, in the compound of formula (1-1) described above, which is an effective component of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention, R in formula (1-1) 1 It can be a straight-chain, branched, or cyclic alkyl group having 1 to 6 carbon atoms. More specifically, examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, cyclopentyl, n-hexyl, cyclohexyl, etc. In the case where the active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention is ropinirole as described below, in formula (1-1), R... 1 It is n-propyl. Therefore, the active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention can also be R in the above formula (1-1). 1 It is a compound with n-propyl groups.

[0139] The compound represented by formula (1-1) above, which is the active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention, may also be 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one. That is, the compound represented by formula (1-1) above may also be ropinirole. The chemical formula of ropinirole is shown in the following formula (1-2).

[0140]

[0141] Ropinirole originally possesses dopamine D2 receptor (D2R) agonist activity on dopamine neurons and was therefore developed as a treatment for Parkinson's disease. In this invention, it is currently unclear whether ropinirole exerts its effect on Huntington's disease through the same intracellular mechanism of action or through a different intracellular mechanism. However, clinical trials as a drug have been completed, and its safety when administered to organisms has been fully confirmed. Thus, because ropinirole is an existing drug, it is possible to rapidly develop Huntington's disease treatment agents or pharmaceutical compositions for Huntington's disease.

[0142] The compound represented by formula (2-1) above, which is the active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention, may be 2-[3-[4-(m-chlorophenyl)-1-piperazinyl]propyl]-s-triazolo[4,3-a]pyridin-3(2H)-one or its hydrochloride salt. That is, the compound represented by formula (2-1) above may be trazodone or its hydrochloride salt. The chemical formula of trazodone is shown in the following formula (2-2).

[0143]

[0144] Trazodone or its hydrochloride salt originally exhibits serotonin (5-HT) reuptake inhibition and has the activity of enhancing serotonin-reducing neuronal function in patients with depression, and has therefore been developed as a treatment for depression / depressive state. In this invention, it is currently unclear whether trazodone acts on Huntington's disease based on the same intracellular mechanism of action or a different intracellular mechanism. However, clinical trials as a medicine have been completed, and its safety when administered to organisms has been well established. Thus, since trazodone or its hydrochloride salt is an existing medicine, it is possible to rapidly develop Huntington's disease treatment agents or pharmaceutical compositions for Huntington's disease.

[0145] The compound represented by formula (3-1) above, which is the active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention, can be N,N-dimethyl-2-[5-(1H-1,2,4-triazol-1-ylmethyl)-1H-indol-3-yl]ethylamine or its benzoate. That is, the compound represented by formula (3-1) above can be rizatriptan or its benzoate. The chemical formula of rizatriptan is shown in the following formula (3-2).

[0146]

[0147] Rizatriptan or its benzoate originally acted on 5-HT1B receptors present in the blood vessels of the skull, selectively constricting extracranial and intracranial arteries that are thought to dilate during migraine attacks. Additionally, it acted on peripheral and central inhibitory 5-HT1D receptors present in the trigeminal nerve, exhibiting activity in inhibiting the release of various peptides (substance P, calcitonin gene-related peptide, etc.), inhibiting vasodilation, dural inflammation, and central pain transmission. Therefore, it was developed as a migraine relief and treatment. In this invention, it is currently unclear whether rizatriptan or its benzoate acts on Huntington's disease based on the same intracellular mechanism of action or a different intracellular mechanism. However, clinical trials as a drug have been completed, and its safety when administered to organisms has been well confirmed. Thus, since rizatriptan or its benzoate is an existing drug, it is possible to rapidly develop Huntington's disease treatment agents or pharmaceutical compositions for Huntington's disease.

[0148] The compound represented by formula (4-1) above, which is an active ingredient in the Huntington's disease treatment agent or pharmaceutical composition for Huntington's disease treatment of the present invention, may be a bromide of 3-dimethylcarbamoyloxy-1-methylpyridinium. That is, the compound represented by formula (4-1) above may be a bromide of pyridostigmine. The chemical formula of pyridostigmine is shown in the following formula (4-2).

[0149]

[0150] Pyridostigmine bromide was originally developed as a treatment for myasthenia gravis primarily by reversibly inhibiting cholinesterase activity at the neuromuscular junction and inhibiting the breakdown of acetylcholine, thereby indirectly enhancing the effects of acetylcholine, and it also possesses acetylcholine-like effects itself. In this invention, it is currently unclear whether pyridostigmine bromide acts on Huntington's disease based on the same intracellular mechanism of action or a different intracellular mechanism. However, clinical trials as a drug have been completed, and its safety in vivo has been well-established. Thus, since pyridostigmine bromide is an existing drug, it is possible to rapidly develop Huntington's disease treatment agents or pharmaceutical compositions for Huntington's disease.

[0151] The active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention may be a salt of any one of the compounds shown in formula (1-1), formula (2-1), formula (3-1), or formula (4-1), or a solvate of any one of the compounds shown in formula (1-1), formula (2-1), formula (3-1), or formula (4-1), or a solvate of a salt of any one of the compounds shown in formula (1-1), formula (2-1), formula (3-1), or formula (4-1).

[0152] When using a salt of any of the compounds represented by formula (1-1), formula (2-1), formula (3-1), or formula (4-1) above in the active ingredient of the Huntington's disease treatment agent or pharmaceutical composition for Huntington's disease treatment of the present invention, there are no particular limitations as long as the salt is pharmaceutically acceptable. Examples include inorganic acid salts such as hydrochloride, sulfate, hydrobromide, nitrate, and phosphate; organic acid salts such as acetate, methanesulfonate, succinate, maleate, fumarate, citrate, and tartrate; alkali metal salts such as sodium and potassium salts; alkaline earth metal salts such as magnesium and calcium salts; metal salts such as aluminum and zinc salts; ammonium salts such as ammonium salts and tetramethylammonium salts; organic amine addition salts such as morpholine and piperidine; and amino acid addition salts such as glycine, phenylalanine, lysine, aspartic acid, and glutamic acid.

[0153] Furthermore, when using any of the compounds or their salts in the Huntington's disease treatment agent or pharmaceutical composition for Huntington's disease treatment of the present invention, there are no particular limitations as long as the solvation is pharmaceutically acceptable, such as hydrates, organic solvations, etc.

[0154] The active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention may also be 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one hydrochloride, i.e. ropinirole hydrochloride.

[0155] The active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention may be 2-[3-[4-(m-chlorophenyl)-1-piperazinyl]propyl]-s-triazolo[4,3-a]pyridine-3(2H)-one hydrochloride, i.e. trazodone hydrochloride.

[0156] The active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention may be N,N-dimethyl-2-[5-(1H-1,2,4-triazol-1-ylmethyl)-1H-indol-3-yl]ethylamine benzoate, i.e. rizatriptan benzoate.

[0157] The active ingredient of the Huntington's disease treatment agent or the pharmaceutical composition for treating Huntington's disease of the present invention may be 3-dimethylcarbamoyloxy-1-methylpyridinium bromide, i.e., pyridostigmine bromide.

[0158] The pharmaceutical composition for treating Huntington's disease of the present invention can be formulated as a pharmaceutical composition, for example, orally in the form of tablets, capsules, elixirs, microcapsules, etc., or non-orally in the form of injections, suppositories, topical skin preparations, etc. More specifically, as topical skin preparations, ointments, patches, and other dosage forms can be included.

[0159] In the Huntington's disease treatment pharmaceutical composition of the present invention, carriers commonly used in pharmaceutical compositions can be used as pharmaceutically acceptable carriers without particular limitation. More specifically, examples include binders such as hydroxypropyl methylcellulose, dextrin, polyethylene glycol 400, gelatin, corn starch, tragacanth gum, and gum arabic; excipients such as lactose hydrate, D-mannitol, starch, crystalline cellulose, and alginate; solvents for injection such as water, ethanol, and glycerin; and adhesives such as rubber-based adhesives and silicone-based adhesives.

[0160] The pharmaceutical composition for treating Huntington's disease of the present invention may include additives. Examples of additives include lubricants such as calcium stearate and magnesium stearate; sweeteners such as sucrose, lactose, saccharin, and maltitol; flavoring agents such as peppermint and red beech oil; stabilizers such as sodium carboxymethyl cellulose, hardened oil, light anhydrous silica, povidone, glyceryl fatty acid esters, benzyl alcohol, and phenol; buffers such as phosphates and sodium acetate; solubilizers such as benzyl benzoate and benzyl alcohol; and colorants such as iron oxide yellow, ferric oxide, iron oxide black, and titanium dioxide.

[0161] The Huntington's disease treatment pharmaceutical composition of the present invention can be formulated by appropriately combining the above-mentioned active ingredient with the above-mentioned pharmaceutically acceptable carrier and additives, and mixing them in a unit dosage form required by generally accepted pharmaceutical practice. The active ingredient of the Huntington's disease treatment pharmaceutical composition of the present invention can be used alone or in combination with two or more ingredients.

[0162] Typically, the appropriate daily dosage of the Huntington's disease treatment pharmaceutical composition of the present invention is the amount of the active ingredient contained in the minimum effective dosage for producing a therapeutic effect. The aforementioned effective minimum dosage depends on various factors including: the activity of the active ingredient contained in the Huntington's disease treatment pharmaceutical composition, the specified fat-soluble / water-soluble functional group modifications, the route of administration, the time of administration, the excretion rate of the specific active ingredient used, the duration of treatment, other drugs, compounds and / or substances used in combination, age, sex, weight, disease, health status and the patient's pre-existing conditions, and other factors known in the medical field. Typically, the dosage of the Huntington's disease treatment pharmaceutical composition of the present invention for a patient is an amount containing approximately 0.0001 to approximately 100 mg / kg body weight of active ingredient per day. The Huntington's disease treatment pharmaceutical composition of the present invention can be administered once daily or in approximately 2 to 4 divided doses.

[0163] In particular, when the active ingredient is a compound represented by formula (1-2), for the dosage of the Huntington's disease treatment pharmaceutical composition of the present invention, for example, if it is a ropinirole sustained-release formulation, it is possible to consider oral administration of 2 mg of the active ingredient once a day, increasing weekly, within the range of no more than 16 to 24 mg of the active ingredient per day.

[0164] In particular, when the active ingredient is a compound represented by formula (2-2), for the dosage of the Huntington's disease treatment pharmaceutical composition of the present invention, for example, 25 mg of the active ingredient can be administered orally three times a day, and the dosage can be increased weekly, within the range of not exceeding 75 to 200 mg of the active ingredient per day.

[0165] In particular, when the active ingredient is a compound represented by formula (3-2), for the dosage of the Huntington's disease treatment pharmaceutical composition of the present invention, for example, 10 mg of the active ingredient can be administered orally once a day, and the dosage can be increased weekly, within the range of not exceeding 10 to 20 mg of the active ingredient per day.

[0166] In particular, when the active ingredient is a compound represented by formula (4-2), for the dosage of the Huntington's disease treatment pharmaceutical composition of the present invention, for example, oral administration of 60 mg of the active ingredient once may be considered, within the range of a maximum daily dose of 180 mg of the active ingredient.

[0167] [Other Implementation Methods]

[0168] In another aspect, the present invention provides a treatment for Huntington's disease, comprising the steps of administering any one of the compounds represented by formula (1-1), formula (2-1), formula (3-1), or formula (4-1), a pharmaceutically acceptable salt thereof, or a solvate thereof to a patient requiring treatment for Huntington's disease. In this aspect of the invention, the same substances as described above can be used as the active ingredient, including any one of the compounds represented by formula (1-1), formula (2-1), formula (3-1), or formula (4-1), a pharmaceutically acceptable salt thereof, or a solvate thereof. Furthermore, in this aspect of the invention, the dosage of the active ingredient can be determined based on suitable dosage studies for each compound, for example, as described above when ropinirole is used as the active ingredient.

[0169] In this invention, any one of the compounds represented by formula (1-1), formula (2-1), formula (3-1), or formula (4-1), a pharmaceutically acceptable salt thereof, or a solvate thereof, is provided for treating Huntington's disease. In this aspect of the invention, the same substances as described above may be used as any one of the compounds represented by formula (1-1), formula (2-1), formula (3-1), or formula (4-1), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0170] The present invention is illustrated below with specific examples. These examples are not intended to limit the invention in any way.

[0171] Example

[0172] Example 1: Screening of compounds for the treatment of Huntington's disease

[0173] In this embodiment, disease-specific iPS cells derived from patients with Huntington's disease are used to differentiate and induce striatal-type intermediate polyspinous neurons (MSNs) that can be used in the screening of compounds for the treatment of Huntington's disease. Compounds that can improve the pathological state of Huntington's disease in these striatal-type MSNs are then screened.

[0174] (1-1) Differentiation / Induction of Striatal Medium-Spiny Neurons (MSNs)

[0175] Huntington's disease is known to present with various pathological states due to an increased number of CAG repeats in the Huntington's gene (HTT gene). In this invention, as disease-specific iPS cells, five patient-derived iPS cells (5 strains) and three healthy-derived iPS cells with varying numbers of CAG repeats are used, based on... Figure 1 The culture method shown induced differentiation into striatal-type MSNs.

[0176] The iPS cells used in this instruction manual are as follows:

[0177] Disease-specific iPS cells

[0178] HD1.CAG180; CAG repeats 180 times.

[0179] HD2.CAG50; CAG repetitions are 50.

[0180] HD3.CAG47; The number of CAG repetitions is 47.

[0181] HD4.CAG50; CAG repetition count is 50 times.

[0182] HD5.CAG42; The CAG repetition count is 42.

[0183] iPS cells from healthy individuals

[0184] ND50018:

[0185] ND50025:

[0186] RC802:

[0187] Specifically, firstly, cell lines containing plasmids linked to promoters induced by doxycycline (DOX) and linked to the genes microRNA9 / 124, ASCL1, BCL2L1, CTIP2, MYT1L, and FOXP1 / 2, which were expressed in the presence of doxycycline (DOX) via the PiggyBac method, were induced for 7 days with doxycycline (CAS number: 24390-14-5) at a final concentration of 2 μg / mL to induce the expression of ASCL1, BCL2L1, CTIP2, MYT1L, and FOXP1 / 2, transcription factors expressed in the initial stage of neuronal differentiation. White, and also contains BDNF (CAS No.: 218441-99-7) at a final concentration of 10 ng / ml, neurotrophic factor-3 (NT-3) at a final concentration of 10 ng / ml (CAS No.: 130939-66-1), valproic acid at a final concentration of 1 μM (CAS No.: 1069-66-5), dibutyryl cAMP (dbcAMP) at a final concentration of 200 μM (CAS No.: 16980-89-5), retinoic acid (RA) at a final concentration of 1 μM (CAS No.: 302-79-4), and B27 (trademark) supplement (Thermo) at a final concentration of 2%. Medium-sized multispinous neurons (MSNs) of the striatal type were induced by culturing them for 7 days in Neurobasal plus (trademark) medium (Thermo Fishcer Scientific A3582901) (medium medium A) from Fischer Scientific (12587010). During this culture process, cytarabine (Ara-C) at a final concentration of 2 μM was added to the medium on days 1 and 3 to stop the proliferation of astrocytes. The medium was changed every 2–3 days.

[0188] Next, the neurotrophic factor-3 (NT-3) at a final concentration of 10 ng / ml (CAS No.: 218441-99-7), at a final concentration of 10 ng / ml (CAS No.: 130939-66-1), at a final concentration of 1 μM valproic acid (CAS No.: 1069-66-5), at a final concentration of 200 μM dibutyryl cAMP (dbcAMP) (CAS No.: 16980-89-5), at a final concentration of 1 μM retinoic acid (RA) (CAS No.: 302-79-4), at a final concentration of 2% B27 supplement (Thermo Fischerscientific, 12587010), and at a final concentration of 0.1% CultureOne supplement (Gibco, A3320201) in Brainphys neuronal culture medium (STEMCELL). Incubate in Technology, ST-05790 (medium B) for 28 to 35 days. Add 1 μM DAPT (CAS No.: 208255-80-5) to the medium only on day 8 when changing the medium. Change the medium every 2–3 days.

[0189] Immunostaining was performed on cells derived from healthy individuals after differentiation induction to investigate the expression of DARPP32 and DRD2, which are MSN markers, and the results are presented in... Figure 2 The results showed that DARPP32 and DRD2 were expressed together with TUJ1, a pan-neuronal cell marker. Figure 3 The study showed that the proportion of DARPP32 and DRD2 positive cells in the obtained neurons was investigated, and the MSNs made from iPS cells derived from Huntington's disease patients had the same proportion as those made from iPS cells derived from healthy individuals. These results indicate that the cells differentiated and induced according to the above steps are striatal-type MSNs.

[0190] Furthermore, it was confirmed whether striatal MSNs derived from Huntington's disease-specific iPS cells after differentiation-induced differentiation exhibited Huntington's disease-specific physiological markers. As Huntington's disease-specific physiological markers, the leakage rate of lactate dehydrogenase (LDH), a marker of neurite cell death, neurite length regression, and HTT aggregate accumulation were measured. LDH leakage rate was measured using a commercially available kit (model "G7891", Promega). Regarding neurite length measurement, immunostaining with TUJ1 antibody was performed on days 7, 14, 21, 28, and 35, and image analysis was performed using this immunostaining as an indicator, quantifying neurite length over time up to day 35 after the start of culture. Regarding the accumulation of HTT aggregates, immunostaining with HTT and polyglutamine antibodies was performed on days 7, 14, 21, 28, and 35. Image analysis was conducted using these immunostains as indicators, and HTT aggregates were quantified over time up to day 35 after the start of culture. All measurements were expressed as a ratio to the average value obtained from MSNs generated from healthy human iPS cells.

[0191] The results clearly showed that, compared with MSNs made from iPS cells derived from healthy individuals, all striatal MSNs derived from Huntington's disease-specific iPS cells exhibited a significantly increased LDH leakage rate. Figure 4 Significant reduction in neurite length ( Figure 5 , Figure 6 ), significant accumulation of HTT aggregates ( Figure 7 , Figure 8 ()( Figure 5 and Figure 7 All results were obtained using cells from day 35. These results indicate that the obtained MSN cells exhibit the pathological characteristics specific to Huntington's disease. Figure 4 , Figure 6 , Figure 8 The asterisks and double angle brackets indicate that there is a significant difference at a significance level of less than 5% or less than 1%, respectively.

[0192] (1-2) Results of wound drug screening using striatal-type medium-sized polyspinous neurons (MSNs)

[0193] Using striatal MSNs derived from Huntington's disease-specific iPS cells induced by differentiation as described in (1-1), drugs that restore the Huntington's disease phenotype were screened from an existing drug library using the reduction in LDH leakage rate, neurite length elongation, and HTT aggregate accumulation as phenotypes shown in (1-1). The determination of LDH leakage rate, neurite length, and HTT aggregate quantification were performed using the same methods as described in Example (1-1). Two types of iPS cells, HD1.CAG180 and HD2.CAG50, were used for screening.

[0194] like Figure 9 As shown, compounds from the existing drug library were added to the culture medium from day 29 to 34, starting from the induction of differentiation, and then screened. The results identified eight drugs that showed promise as therapeutic agents for Huntington's disease. The improvement rate (%) of Huntington's disease phenotype when these drugs were added to the culture medium was calculated.

[0195] The improvement rate (%) of Huntington's disease phenotype was calculated for each parameter using the following formula (1):

[0196] Improvement rate (%) = (AB) / (AC) × 100…(1)

[0197] In formula (1),

[0198] A represents the measured value of striatal-type medium-sized polyspinous neurons (MSNs) induced by differentiation of iPS cells derived from HD patients in the absence of drugs.

[0199] B represents the measured value of striatal-type intermediate-sized polyspinous neurons (MSNs) induced by differentiation of iPS cells derived from HD patients in the presence of the drug.

[0200] C represents the measured value of striatal-type intermediate-sized multispinous neurons (MSNs) induced by differentiation of iPS cells from healthy individuals in the absence of drugs.

[0201] It should be noted that the proportions of each phenotype—LDH leakage, neurite retraction, and HTT aggregate accumulation—were the highest at 35%, 35%, and 30%, respectively, totaling 100%.

[0202] The eight drugs obtained above were added to the culture medium of striatal-type medium-sized polyspinous neurons (MSNs) induced by differentiation of iPS cells (HD1.CAG180 and HD2.CAG50) derived from HD patients at final concentrations of 0.01 μM, 0.1 μM, 1 μM, and 10 μM, and the results are shown in Figure 10. These results show that, when the improvement rate (%) of Huntington's disease phenotypes is converted to the combined proportion of LDH leakage, neurite regression, and HTT aggregate accumulation as 100%, ropinirole, trazodone, rizatriptan, and pyridostigmine exhibited high neuroprotective effects in the range of 0.01 μM–10 μM.

[0203] Based on these results, ropinirole was selected from these compounds for further investigation. Immunostaining of cells also showed that ropinirole improves neurite retraction (…). Figure 11 (using HD2.CAG50 as cells) and its effect on improving HTT aggregates ( Figure 12 (HD1.CAG180 cells were used). It should be noted that Table 1 shows the improvement rate of ropinirole on each phenotype in the two cell types (HD1.CAG180 and HD2.CAG50).

[0204] [Table 1]

[0205]

[0206] Example 2: The effects of ropinirole

[0207] In this embodiment, functional analysis was performed on striatal-type medium-sized polyspinous neurons (MSNs) made from disease-specific iPS cells derived from cells of Huntington's disease patients, obtained from ropinirole in Example 1 (1-2).

[0208] (2-1) Steps for treating cells with ropinirole

[0209] The timing of adding ropinirole to the culture, such as Figure 13 The change shown was from day 29 to day 36 from the start of differentiation induction. Otherwise, the efficacy of ropinirole was evaluated in the same manner as in (1-2) of Example 1.

[0210] Specifically, ropinirole was added to the culture medium of striatal-type medium-sized polyspinous neurons (MSNs) induced by iPS cell differentiation from Huntington's disease patients with HTT gene mutations (HD1.CAG180, HD2.CAG50, HD3.CAG47) on days 29, 31, and 34, and assays were performed on day 36. Ropinirole was added to the culture medium at final concentrations of 10 nM, 100 nM, and 1000 nM.

[0211] (2-2) Neuroprotective effects of ropinirole

[0212] As a parameter of neuroprotective effect, the LDH leakage rate induced by ropinirole administration was measured. The LDH leakage rate was measured in the same manner as in (1-2) of Example 1. The results are shown in... Figure 14 and Figure 15 .

[0213] Figure 14 This is a graph showing the results of measuring LDH leakage rates for striatal-type medium-sized polyspinous neurons (MSNs) induced by differentiation of iPS cells (HD1.CAG180, HD2.CAG50, HD3.CAG47) derived from Huntington's disease patients with different CAG repeat numbers, when various concentrations (10 nM, 100 nM, 1000 nM) of ropinirole were added to the culture medium. Figure 14 The vertical axis represents the LDH leakage rate (relative value) when various concentrations of ropinirole are added, based on the LDH leakage value without the addition of ropinirole. The horizontal axis represents the results of adding ropinirole to the culture medium at any concentration of 10 nM, 100 nM, or 1000 nM. Figure 14 The asterisk (*) indicates a significant difference at a significance level of less than 5%, and the double-digit symbol (**) indicates a significant difference at a significance level of less than 1%.

[0214] Figure 15 This demonstrates that for intermediate-sized polyspinous neurons (MSNs) of the striatum type induced by differentiation of iPS cells (HD1.CAG180, HD2.CAG50, HD5.CAG42) derived from Huntington's disease patients with different CAG repeat numbers, compared to... Figure 14 The experiment provides more detailed graphs showing the LDH leakage rate determination results when various concentrations (0.1 nM, 1 nM, 10 nM, 30 nM, 100 nM, 300 nM) of ropinirole were added to the culture medium. Figure 15 The vertical axis represents the LDH leakage rate (relative value) when various concentrations of ropinirole are added, based on the LDH leakage value without the addition of ropinirole. The horizontal axis represents the result of adding ropinirole to the culture medium at any concentration of 0.1 nM, 1 nM, 10 nM, 30 nM, 100 nM, or 300 nM. Figure 15 The asterisk (*) indicates a significant difference at a significance level of less than 5%, and the double-digit symbol (**) indicates a significant difference at a significance level of less than 1%.

[0215] Figure 14 and Figure 15 The results showed that even on days 29–36 from the start of differentiation induction, the addition of ropinirole confirmed a neuroprotective effect (reduction in LDH leakage). Furthermore, Figure 14 The EC50 of ropinirole in the medium is below 10 nM (calculated using ImageJ and Excel) and Figure 15 The IC50 of ropinirole in the study was 64.2 nM (calculated using ImageJ and Excel), demonstrating its ability to exert neuroprotective effects at very low doses.

[0216] (2-3) Improvement of HTT aggregate accumulation by ropinirole

[0217] For the accumulation of HTT aggregates, a physiological marker specific to Huntington's disease, following differentiation-induced differentiation of intermediate-sized polyspinous neurons (MSNs) derived from striatal iPS cells (a disease-specific iPS cell), the change in HTT aggregate accumulation rate induced by ropinirole administration was measured as a parameter for the improvement of HTT aggregate accumulation. The HTT aggregate accumulation rate was measured in the same manner as in Example 1. The results are presented in... Figure 16 and Figure 17 .

[0218] Figure 16 The figure shows the following results after immunostaining: For striatal-type medium-sized polyspinous neurons (MSNs) induced from the differentiation of iPS cells (HD1.CAG180, HD2.CAG50, HD5.CAG42) derived from Huntington's disease patients with different CAG repeat numbers, the addition of various concentrations (10 nM, 30 nM, 100 nM, 300 nM) of ropinirole to the culture medium improved the accumulation of HTT aggregates. Figure 16 The arrows indicate the accumulation sites of HTT aggregates, and the horizontal axis represents the results of adding ropinirole to the culture medium at any concentration of 10 nM, 30 nM, 100 nM, or 300 nM.

[0219] Figure 17 It is Figure 16 The results are presented in numerical charts. Figure 17 The vertical axis represents the HTT aggregate accumulation rate (relative value) when various concentrations of ropinirole are added, based on the HTT number under the condition without ropinirole. The horizontal axis represents the result of adding ropinirole to the culture medium at any concentration of 10 nM, 30 nM, 100 nM, or 300 nM. Figure 17The asterisk (*) indicates a significant difference at a significance level of less than 5%, and the double-digit symbol (**) indicates a significant difference at a significance level of less than 1%.

[0220] according to Figure 16 and Figure 17 The results showed that, from day 29 to 36 of differentiation induction, the addition of ropinirole also confirmed an improvement in HTT aggregate accumulation (a reduction in HTT aggregate accumulation rate). Furthermore, ropinirole's IC50 = 40.5 nM (calculated using ImageJ and Excel) demonstrated that it could exert its effect in improving HTT aggregate accumulation at very low doses.

[0221] (2-4) Neuroprotective effects of ropinirole compared to the same active ingredient

[0222] Ropinirone possesses dopamine D2 receptor (D2R) agonist activity on dopamine neurons; therefore, the neuroprotective effects of bromocriptine, also a D2 receptor agonist, were compared with those of ropinirone. The LDH leakage rate induced by ropinirone administration was measured as a parameter of neuroprotective effect. The LDH leakage rate was measured in the same manner as in Example 1. The results are presented below. Figure 18 .

[0223] Figure 18 This is a graph showing the results of LDH leakage in striatal-type medium-sized polyspinous neurons (MSNs) induced by differentiation of iPS cells (HD1.CAG180, HD2.CAG50, HD5.CAG42) from Huntington's disease patients, measured in culture media with various concentrations of ropinirole and bromocriptine (a D2 receptor agonist). Figure 18 The vertical axis represents the LDH leakage rate (relative value) when various concentrations of ropinirole were added, based on the LDH leakage value without the addition of ropinirole. The horizontal axis represents the results when ropinirole was added to the culture medium at any of the following concentrations: 1 nM, 30 nM, 300 nM, or bromocriptine at any of the following concentrations: 1 nM, 30 nM, 300 nM. Figure 15 The asterisk (*) indicates a significant difference at a significance level of less than 5%, and the double-digit symbol (**) indicates a significant difference at a significance level of less than 1%.

[0224] The results showed that neuroprotective effects (reduction of LDH leakage) were confirmed by the addition of ropinirole from days 29 to 36, starting from the start of differentiation induction, but no neuroprotective effects were confirmed by bromocriptine. These results indicate that ropinirole, a D2 receptor agonist, exerts a neuroprotective effect compared to bromocriptine with the same agonist.

[0225] (2-5) Neuroprotective effect of ropinirole against glutamate-induced cell death

[0226] It is known that when glutamate neurostimulation is excessive, nerve cells become overexcited and unable to tolerate the stimulation, leading to cell death. This phenomenon is called glutamate neurotoxicity and is considered a cause of various neurodegenerative diseases. To confirm whether ropinirole has a neuroprotective effect against glutamate neurotoxicity, the LDH leakage rate induced by ropinirole administration was measured as a parameter of neuroprotective effect. The LDH leakage rate was measured in the same manner as in Example 1. The results are shown in... Figure 19 .

[0227] Figure 19 This is a graph showing the results of LDH leakage measurements in striatal-type medium-sized polyspinous neurons (MSNs) induced by differentiation of iPS cells (HD2.CAG50) derived from Huntington's disease patients, with the addition of glutamate and various concentrations of ropinirole to the culture medium. Figure 19 The vertical axis represents the LDH leakage rate (relative value) when various concentrations of ropinirole are added to cells with 50 μM glutamate, based on the LDH leakage value under conditions where neither glutamate nor ropinirole is added. The horizontal axis represents the results when ropinirole is added to the culture medium at any concentration of 10 nM, 100 nM, or 1000 nM. Figure 19 The "+" on the horizontal axis indicates that the culture medium has been supplemented with 50 μM glutamic acid. Figure 19 The asterisk (*) indicates a significant difference at a significance level of less than 5%, and the double-digit symbol (**) indicates a significant difference at a significance level of less than 1%.

[0228] The results showed that, from day 29 to 36 of differentiation induction, neuronal death was observed with the addition of glutamate, but neuroprotective effects (reduction of LDH leakage) were confirmed with the addition of ropinirole. These results indicate that ropinirole exerts a neuroprotective effect against overstimulation of glutamate receptors.

[0229] (2-6) Neuroprotective effects of ropinirole using various HD strains

[0230] As a parameter of the neuroprotective effect induced by ropinirole administration, the LDH leakage rate induced by ropinirole administration was measured, and the improvement effect of adding ropinirole on the LDH leakage rate was studied based on the change in LDH leakage rate. The calculation of the LDH leakage improvement rate was performed in the same manner as in (1-2) of Example 1. The results are shown in... Figure 20 .

[0231] Figure 20The results show the LDH leakage rate of striatal-type medium-sized polyspinous neurons (MSNs) induced by differentiation of iPS cells (HD1.CAG180, HD2.CAG50, HD3.CAG47, HD4.CAG50, HD5.CAG42) derived from Huntington's disease patients when 300 nM ropinirole was added to the culture medium. The graph shows the improvement rate of LDH leakage after ropinirole administration. Figure 20 The horizontal axis represents the LDH leakage improvement rate (relative value) from the LDH leakage value before ropinirole administration (0 nM) to the LDH leakage value after ropinirole administration (300 nM), with the LDH leakage value of healthy human samples as the baseline (100%). The vertical axis represents the results for each cell in which ropinirole was added to the culture medium (0 nM or 300 nM). Figure 20 The asterisk (*) indicates a significant difference at a significance level of less than 5%, and the double-digit symbol (**) indicates a significant difference at a significance level of less than 1%.

[0232] As a result, neuroprotective effects (improved LDH leakage) were confirmed even on days 29–36 from the start of differentiation induction with the addition of ropinirole. These results demonstrate that the neuroprotective effects of ropinirole are effective in treating patients with a wide range of HTT CAG repeats.

[0233] Industrial availability

[0234] According to the present invention, a therapeutic agent for Huntington's disease and a composition for treating Huntington's disease can be provided. The Huntington's disease therapeutic agent or composition of the present invention can be used not only to treat Huntington's disease but also to prevent its onset. Furthermore, by analyzing the pharmacodynamic mechanism of the Huntington's disease therapeutic agent of the present invention on striatal-type intermediate polyspinous neurons (MSNs) differentiated from iPS cells derived from Huntington's disease patients, the pathological mechanisms of Huntington's disease can be elucidated.

Claims

1. A Huntington's disease treatment agent comprising a compound represented by formula (1-1), a compound represented by formula (2-1), a compound represented by formula (3-1), a compound represented by formula (4-1), a pharmaceutically acceptable salt thereof, or a solvate thereof. In equation (1-1), R 1 Each of these can independently represent an alkyl group or a 4-hydroxyphenylethyl group with 1 to 6 carbon atoms, where n represents an integer from 1 to 3. In equation (2-1), R 21 For H or -CH3, R 22 Choose from the group consisting of H, alkyl groups with 1 to 6 carbon atoms, alkoxy groups with 1 to 6 carbon atoms, and halogens. In equation (3-1), Y represents N or A 2 -C, Z represents N or CH. m can be 0, 1, 2, or 3. B represents O, S, or NR. 33 , A 1 and A 2 Independently representing an alkyl group having 1 to 6 carbon atoms that is optionally substituted, an alkenyl group having 2 to 6 carbon atoms that is optionally substituted, an alkynyl group having 2 to 6 carbon atoms that is optionally substituted, a cycloalkyl group having 3 to 7 carbon atoms that is optionally substituted, an aryl group having 1 to 6 carbon atoms that is optionally substituted, a heterocycloalkyl group having 3 to 7 carbon atoms that is optionally substituted, a heteroaryl group having 1 to 6 carbon atoms that is optionally substituted, or a heteroaryl group having 1 to 6 carbon atoms that is optionally substituted, or H, halogen, cyano, trifluoromethyl, an alkoxy group having 1 to 6 carbon atoms, an alkylthio group having 1 to 6 carbon atoms, or -NR x R y , in, R x and R y Independently representing H, hydrocarbon group, or heterocyclic group, or R x and R y Together they represent alkylene groups having 2 to 6 carbon atoms. R 31 -CH2CHR 34 NR 35 R 36 Or the following formula: R 32 R 33 R 34 R 35 R 36 and R 37 Independently representing H or an alkyl group having 1 to 6 carbon atoms, In equation (4-1), R 41 and R 42 Selected from alkyl groups having 1 to 6 carbon atoms, aryl groups having 6 to 10 carbon atoms, and alkyl groups having 1 to 6 carbon atoms substituted with aryl groups having 6 to 10 carbon atoms. R 43 Selected from alkyl groups having 1 to 6 carbon atoms, M is selected from halogens and alkyl sulfates.

2. The Huntington's disease treatment agent according to claim 1, wherein, In equation (1-1), n ​​is 2.

3. The Huntington's disease treatment agent according to claim 1 or 2, wherein, In equation (1-1), R 1 It is n-propyl.

4. The Huntington's disease treatment agent according to claim 1 or 2, wherein, The compound represented by formula (1-1) is the same as the compound represented by formula (1-2) below: 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indol-2-one.

5. The Huntington's disease treatment agent according to claim 4, wherein, The pharmaceutically acceptable salt of the compound represented by formula (1-1) is the hydrochloride salt of the compound represented by formula (1-2), namely 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one hydrochloride.

6. The Huntington's disease treatment agent according to claim 1 or 2, wherein, The compound represented by formula (2-1) is the same as the compound represented by formula (2-2) below: 2-[3-[4-(m-chlorophenyl)-1-piperazinyl]propyl]-s-triazolo[4,3-a]pyridine-3(2H)-one.

7. The Huntington's disease treatment agent according to claim 6, wherein, The pharmaceutically acceptable salt of the compound represented by formula (2-1) is the hydrochloride salt of the compound represented by formula (2-2), namely 2-[3-[4-(m-chlorophenyl)-1-piperazinyl]propyl]-s-triazolo[4,3-a]pyridine-3(2H)-one hydrochloride.

8. The Huntington's disease treatment agent according to claim 1 or 2, wherein, The compound represented by formula (3-1) is the same as the compound represented by formula (3-2) below: N,N-Dimethyl-2-[5-(1H-1,2,4-triazol-1-ylmethyl)-1H-indol-3-yl]ethylamine.

9. The Huntington's disease treatment agent according to claim 8, wherein, The pharmaceutically acceptable salt of the compound represented by formula (3-1) is the benzoate of the compound represented by formula (3-2), namely N,N-dimethyl-2-[5-(1H-1,2,4-triazol-1-ylmethyl)-1H-indol-3-yl]ethylamine benzoate.

10. The Huntington's disease treatment agent according to claim 1 or 2, wherein, The compound represented by formula (4-1) is the same as the compound represented by formula (4-2) below: 3-Dimethylcarbamoyloxy-1-methylpyridinium bromide.

11. A pharmaceutical composition for treating Huntington's disease, comprising, as an active ingredient, a compound of formula (1-1), a compound of formula (2-1), a compound of formula (3-1), a compound of formula (4-1), a pharmaceutically acceptable salt thereof, or a solvate thereof. In equation (1-1), R 1 Each of these can independently represent an alkyl group or a 4-hydroxyphenylethyl group with 1 to 6 carbon atoms, where n represents an integer from 1 to 3. In equation (2-1), R 21 For H or -CH3, R 22 Choose from the group consisting of H, alkyl groups with 1 to 6 carbon atoms, alkoxy groups with 1 to 6 carbon atoms, and halogens. In equation (3-1), Y represents N or A 2 -C, Z represents N or CH. m can be 0, 1, 2, or 3. B represents O, S, or NR. 33 , A 1 and A 2 Independently representing an alkyl group having 1 to 6 carbon atoms that is optionally substituted, an alkenyl group having 2 to 6 carbon atoms that is optionally substituted, an alkynyl group having 2 to 6 carbon atoms that is optionally substituted, a cycloalkyl group having 3 to 7 carbon atoms that is optionally substituted, an aryl group having 1 to 6 carbon atoms that is optionally substituted, a heterocycloalkyl group having 3 to 7 carbon atoms that is optionally substituted, a heteroaryl group having 1 to 6 carbon atoms that is optionally substituted, or a heteroaryl group having 1 to 6 carbon atoms that is optionally substituted, or H, halogen, cyano, trifluoromethyl, an alkoxy group having 1 to 6 carbon atoms, an alkylthio group having 1 to 6 carbon atoms, or -NR x R y , in, R x and R y Independently representing H, hydrocarbon group, or heterocyclic group, or R x and R y Together they represent alkylene groups having 2 to 6 carbon atoms. R 31 -CH2CHR 34 NR 35 R 36 Or the following formula: R 32 R 33 R 34 R 35 R 36 and R 37 Independently representing H or an alkyl group having 1 to 6 carbon atoms, In equation (4-1), R 41 and R 42 Selected from alkyl groups having 1 to 6 carbon atoms, aryl groups having 6 to 10 carbon atoms, and alkyl groups having 1 to 6 carbon atoms substituted with aryl groups having 6 to 10 carbon atoms. R 43 Selected from alkyl groups having 1 to 6 carbon atoms, M is selected from halogens and alkyl sulfates.

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