Neuronal degeneration inhibitors

Abstract

The purpose of the present invention is to provide a nerve cell degeneration inhibitor. The present invention pertains to a nerve cell degeneration inhibitor comprising a compound represented by formula (I) [each symbol in the formula is as described in the specification] or a salt thereof.

Description

[Technical Field]

[0001] The present invention relates to an agent (pharmaceutical, pharmaceutical composition) containing a compound useful for treating motor neuron disease or dementia.

[0002] (Background of the invention) Eukaryotic genomes contain various repeat sequences, but in recent years, a 6-base repeat sequence called "GGGGCC" (sometimes referred to as the G4C2 repeat) located in the uncoding region of the C9orf72 gene has attracted attention as a link between amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). ALS is a representative motor neuron disease characterized by selective degeneration of upper and lower motor neurons, with approximately 10% of patients having sporadic cases and approximately 90% having familial cases. While extensive research has focused on gene mutations found in familial patients, recent studies have identified abnormal G4C2 repeat elongation in the C9orf72 gene as the most frequently observed gene abnormality in both familial and sporadic ALS patients. Meanwhile, FTD is the second most common type of dementia in those under 65 years of age, characterized by degeneration of the frontal and temporal lobes of the brain. Furthermore, abnormal G4C2 repeat elongation in the C9orf72 gene has been reported as the most frequently observed gene abnormality in FTD as well (Non-patent Literature 1, 2). RNA transcribed from the C9orf72 gene, in which the G4C2 repeat is abnormally elongated, is known to readily aggregate and form nuclear aggregates (hereinafter referred to as RNA foci) by involving nuclear proteins (Non-Patent Documents 1 and 2). Furthermore, this RNA is translated into dipeptide repeat proteins (hereinafter referred to as DPRs) by repeat-associated non-ATG translation (hereinafter referred to as RAN translation), and DPRs are known to form inclusion bodies within cells (Non-Patent Documents 3, 4, and 5). RNA foci and DPRs are considered to be major causes of neurodegeneration in this disease due to their cytotoxicity (Non-Patent Document 6), and it is believed that effectively suppressing their generation could prevent the onset of the disease or effectively suppress the progression of the disease after onset. To date, various model systems have been used to search for genes that can suppress the formation of RNA foci and / or DPRs. For example, screening using a Drosophila model that expresses abnormally elongated G4C2 repeats in the compound eye identified FUS (the gene responsible for ALS6) as a gene that suppresses RAN translation and compound eye neurodegeneration when overexpressed (Patent Document 1). In addition, antisense oligonucleotides against the C9orf72 gene have been developed and have been reported to effectively suppress neuronal cell death by inhibiting RNA foci formation (for example, Non-Patent Document 7). However, no small molecule compounds that can effectively inhibit the generation of RNA foci and / or DPRs caused by G4C2 repeats have yet been reported. [Prior art documents] [Patent Documents]

[0003] [Patent Document 1] Japanese Patent Publication No. 2018-193309 [Non-patent literature]

[0004] [Non-Patent Document 1] DeJesus-Hernandez, M. et al., Neuron 72, 245-56 (2011) [Non-Patent Document 2] Renton, AE et al., Neuron 72, 257-68 (2011) [Non-Patent Document 3] Ash,PEet al.,Neuron 77,639-46(2013) [Non-Patent Document 4] Mori,K.et al.,Science 339,1335-8(2013) [Non-Patent Document 5] Zu,T.et al.,Proc Natl Acad Sci USA 110,E4968-77(2013) [Non-Patent Document 6] J.Chew et al.,Science,348,1151-1154(2015) [Non-Patent Document 7] D. Sareen et al.,Sci.Transl.Med.,5,208ra149,doi:10.1126 / scitranslmed.3007529(2013) [Overview of the project] [Problems that the invention aims to solve]

[0005] The object of the present invention is to find compounds that can effectively suppress the production of RNA foci and DPRs, which are considered to be the causes of neurodegeneration in motor neuron diseases and dementias such as FTD, and to provide agents (pharmaceuticals, pharmaceutical compositions) that can be used as preventive or therapeutic agents for said diseases. [Means for solving the problem]

[0006] To solve the above problems, the inventors first discovered that motor neurons differentiated from iPS cells derived from ALS patients with abnormally elongated G4C2 repeats by overexpression of the Lhx3, Ngn2, and Isl1 genes spontaneously generate RNA foci and DPRs, leading to neurodegeneration. The differentiation induction method using the expression of these three genes is a method developed by the inventors that can induce differentiation into motor neurons in a short period of time with high synchronicity (WO 2014 / 148646, and K. Imamura et al, Science Translational Medicine 2017, 9, eaaf3962). Then, using the aforementioned motor neurons, we screened for small molecule compounds and found that seven compounds have excellent inhibitory effects on RNA foci and DPRs production, meaning they can be used as preventive or therapeutic agents for motor neuron diseases and / or dementia caused by the abnormal repeat extensions, thus completing the present invention.

[0007] In other words, the present invention is as follows: [1] Equation (I)

[0008] [ka]

[0009] [In the formula, R 1 is equation (a-1) or (a-2)

[0010] [ka]

[0011] (In the formula, R 11 and R 12 These are, independently, hydrogen atoms or C 1-6 It shows an alkyl group, R 13 is a hydrogen atom, a cyano group, C 1-6 Alkyl-carbonyl group or C1-6 represents an alkoxy-carbonyl group, R 14 is a C 1-6 alkyl group, a C 3-8 cycloalkyl group or a C 6-14 aryl group. ) represents a group represented by R 2 is of formula (b-1) to (b-3)

[0012]

Chemical formula

[0013] (wherein, R 21 is a C 1-6 alkyl group, a C 6-14 aryl group optionally substituted with a halogen atom, or a C 7-16 aralkyl group optionally substituted with a halogen atom, R 22 each independently represents a halogen atom, a cyano group, a C 1-6 alkyl group or a C 1-6 alkoxy group, n represents 0, 1 or 2. ) represents a group represented by R 3 each independently represents a halogen atom, a cyano group, a C 1-6 alkyl group or a C<​​​​​​​​​​​​​​​​​​​​​​​ R 14 However, C 1-6 It is an alkyl group, R 21 However, C 1-6 C may be substituted with alkyl or halogen atoms. 7-16 It is an aralkyl group, R 22 However, it is a halogen atom, n is 0 or 1, m is 0 and L is a methylene group. The neuronal degeneration inhibitor described in [1] above.

[0015] [3] A compound represented by formula (I), or a salt thereof, (1) 4-(4-fluoro-2-methoxyphenyl)-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (2) 1-(3-{[4-(2-methoxyphenyl)-1,3,5-triazine-2-yl]amino}phenyl)methanesulfonamide, (3) 1-(3-{[4-(4-chloro-2-methoxyphenyl)-1,3,5-triazine-2-yl]amino}phenyl)methanesulfonamide, (4) 1-[3-({4-[2-(benzyloxy)phenyl]-1,3,5-triazine-2-yl}amino)phenyl]methanesulfonamide, (5) 4-{2-[(3,4-dichlorophenyl)methoxy]phenyl}-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (6) Ethyl {[(3-{[4-(2,3-dihydro-1,4-benzodioxin-5-yl)-1,3,5-triazine-2-yl]amino}phenyl)methyl](methyl)oxo-λ 6 -Sulfanylidene}calbamate, (7) Ethyl {[(3-{[4-(2,3-dihydro-1-benzofuran-7-yl)-1,3,5-triazine-2-yl]amino}phenyl)methyl](methyl)oxo-λ6 -Sulfanylidene}calbamate, and neuronal degeneration inhibitors selected from salts thereof, as described in [1] above.

[0016] [4] A neurodegenerative inhibitor as described in any of [1] to [3] above, used as a preventive or therapeutic agent for motor neuron disease or dementia. [5] The neuronal degeneration inhibitor described in [4] above, wherein the motor neuron disease or dementia is a motor neuron disease or dementia accompanied by abnormal elongation of hexanucleotide repeats. [6] The neuronal degeneration inhibitor described in [4] or [5] above, wherein the motor neuron disease is amyotrophic lateral sclerosis. [7] The neurodegeneration inhibitor according to [4] or [5] above, wherein the dementia is frontotemporal dementia. [8] A method for inhibiting neuronal degeneration in a mammal, comprising administering an effective amount of any of the compounds or salts thereof described in any of [1] to [3] above to the mammal. [9] A method for preventing or treating a neurodegenerative disease in a mammal, comprising administering an effective amount of any of the compounds or salts thereof described in any of [1] to [3] above to the mammal.

[10] A compound or salt thereof described in any of [1] to [3] above, for the prevention or treatment of neurodegenerative diseases.

[11] Use of any of the compounds or salts thereof described in [1] to [3] above for the manufacture of agents for the prevention or treatment of neurodegenerative diseases. [Effects of the Invention]

[0017] According to the present invention, an agent is provided that is excellent at suppressing the production of RNA foci and DPRs in neurodegenerative diseases accompanied by abnormal G4C2 repeat elongation of the C9orf72 gene, and can be used for the prevention or treatment of said diseases. [Brief explanation of the drawing]

[0018] [Figure 1]This is a fluorescence microscope image of motor neurons differentiated from ALS7 (C9orf72) cells, in which RNA foci were visualized using a fluorescent probe.

[0019] (Detailed description of the invention) The present invention will be described in detail below. The present invention provides a neuronal degeneration inhibitor containing a compound represented by the following formula (I), or a salt thereof (hereinafter sometimes referred to as "compound (I)") as an active ingredient.

[0020] [ka]

[0021] [In the formula, R 1 is equation (a-1) or (a-2)

[0022] [ka]

[0023] (In the formula, R 11 and R 12 These are, independently, hydrogen atoms or C 1-6 It shows an alkyl group, R 13 is a hydrogen atom, a cyano group, C 1-6 Alkyl-carbonyl group or C 1-6 It shows an alkoxy-carbonyl group, R 14 C 1-6 Alkyl alkyl group, C 3-8 Cycloalkyl groups or C 6-14 (This indicates an aryl group.) The base represented by R 2 This is the equation (b-1) to (b-3)

[0024] [ka]

[0025] (In the formula, R 21 C 1-6 C may be substituted with alkyl or halogen atoms. 6-14 C may be substituted with an aryl group or a halogen atom. 7-16 It shows an aralkyl group, R 22 These are, independently, a halogen atom, a cyano group, and C 1-6 Alkyl or C 1-6 It shows an alkoxy group, n represents 0, 1, or 2. The base represented by R 3 These are, independently, a halogen atom, a cyano group, and C 1-6 Alkyl or C 1-6 It shows an alkoxy group, m represents 0, 1, or 2. L is C 1-3 [Indicates an alkylene group.]

[0026] The definitions of each substituent used in this specification are described in detail below. Unless otherwise specified, each substituent has the following definitions. In this specification, examples of "halogen atoms" include fluorine, chlorine, bromine, and iodine. In this specification, "C 1-6 Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, 1-ethylpropyl, hexyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, and 2-ethylbutyl.

[0027] In this specification, "C 3-8Examples of "cycloalkyl groups" include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl, and adamantyl. In this specification, "C 6-14 Examples of "aryl groups" include phenyl, 1-naphthyl, 2-naphthyl, 1-anthryl, 2-anthryl, and 9-anthryl. In this specification, "C 7-16 Examples of "aralkyl groups" include benzyl, phenethyl, naphthylmethyl, and phenylpropyl.

[0028] In this specification, "C 1-6 Examples of "alkoxy groups" include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, and hexyloxy. In this specification, "C 1-6 Examples of alkyl-carbonyl groups include acetyl, propanoyl, butanoyl, 2-methylpropanoyl, pentanol, 3-methylbutanoyl, 2-methylbutanoyl, 2,2-dimethylpropanoyl, hexanoyl, and heptanol. In this specification, "C 1-6 Examples of "alkoxy-carbonyl groups" include methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl, and hexyloxycarbonyl.

[0029] In this specification, "C 1-3 Examples of alkylene groups include -CH2-, -(CH2)2-, -(CH2)3-, -CH(CH3)-, -CH2CH(CH3)-, -CH(CH3)CH2-, -C(CH3)2-, and -CH(C2H5)-.

[0030] The definitions of each symbol in equation (I) are described in detail below. R 1 is equation (a-1) or (a-2)

[0031] [ka]

[0032] This indicates the group represented by [the symbol]. Here, R 11 and R 12 These are, independently, hydrogen atoms or C 1-6 R indicates an alkyl group. 13 is a hydrogen atom, a cyano group, C 1-6 Alkyl-carbonyl group or C 1-6 It shows an alkoxy-carbonyl group, R 14 C 1-6 Alkyl alkyl group, C 3-8 Cycloalkyl groups or C 6-14 This indicates an aryl group. R 11 and R 12 Preferably, both are hydrogen atoms. R 13 Preferably, a hydrogen atom or C 1-6 The group is an alkoxy-carbonyl group (e.g., ethoxycarbonyl), and is particularly preferably a hydrogen atom. R 14 Preferably, C 1-6 The alkyl group is (e.g., methyl), and is particularly preferably a methyl group. R 1 Preferably, it is a group represented by formula (a-2).

[0033] R 2 This is the equation (b-1) to (b-3)

[0034] [ka]

[0035] This indicates the group represented by [the symbol]. Here, R21 is C 1-6 an alkyl group, a C optionally substituted with a halogen atom 6-14 aryl group, or a C optionally substituted with a halogen atom 7-16 aralkyl group, and R 22 each independently represents a halogen atom, a cyano group, a C 1-6 alkyl group or a C 1-6 alkoxy group, and n represents 0, 1 or 2.

[0036] R 21 is preferably a C 1-6 alkyl group (e.g., methyl), or a C optionally substituted with a halogen atom (e.g., chlorine atom) 7-16 aralkyl group (e.g., benzyl), more preferably a C 1-6 alkyl group (e.g., methyl), and particularly preferably a methyl group. R 22 is preferably a halogen atom (e.g., fluorine atom, chlorine atom), and particularly preferably a fluorine atom. n is preferably 0 or 1, and particularly preferably 1. R 2 are preferably groups represented by formula (b-1).

[0037] R 3 each independently represents a halogen atom, a cyano group, a C 1-6 alkyl group or a C 1-6 alkoxy group. m represents 0, 1 or 2. m is preferably 0.

[0038] L is a C 1-3 alkylene group. L is preferably a methylene group.

[0039] Preferred embodiments of the compound (I) include the following compounds. R 1 is a group represented by formula (a-1) or (a-2), R11 and R 12 However, both are hydrogen atoms, R 13 However, hydrogen atoms or C 1-6 It is an alkoxy-carbonyl group (e.g., ethoxycarbonyl), R 14 However, C 1-6 It is an alkyl group (e.g., methyl), R 2 However, it is a base represented by equations (b-1) to (b-3), R 21 However, C 1-6 C may be substituted with an alkyl group (e.g., methyl) or a halogen atom (e.g., chlorine). 7-16 It is an aralkyl group (e.g., benzyl), R 22 However, these are halogen atoms (e.g., fluorine atoms, chlorine atoms), n is 0 or 1, m is 0 and L is a methylene group. Compound (I).

[0040] Specific examples of compound (I) include the following compounds: (1) 4-(4-fluoro-2-methoxyphenyl)-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine (compound number 1), (2) 1-(3-{[4-(2-methoxyphenyl)-1,3,5-triazine-2-yl]amino}phenyl)methanesulfonamide (compound number 2), (3) 1-(3-{[4-(4-chloro-2-methoxyphenyl)-1,3,5-triazine-2-yl]amino}phenyl)methanesulfonamide (compound number 3), (4) 1-[3-({4-[2-(benzyloxy)phenyl]-1,3,5-triazine-2-yl}amino)phenyl]methanesulfonamide (compound numbers 4a, 4b), (5) 4-{2-[(3,4-dichlorophenyl)methoxy]phenyl}-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine (compound number 5), (6) Ethyl {[(3-{[4-(2,3-dihydro-1,4-benzodioxin-5-yl)-1,3,5-triazine-2-yl]amino}phenyl)methyl](methyl)oxo-λ 6 -Sulfanylidene carbamate (compound number 6), (7) Ethyl {[(3-{[4-(2,3-dihydro-1-benzofuran-7-yl)-1,3,5-triazine-2-yl]amino}phenyl)methyl](methyl)oxo-λ 6 -Sulfanylidene carbamate (compound number 7), and their salts.

[0041] If compound (I) is a salt, a pharmacologically acceptable salt is preferred, and examples of such salts include salts with inorganic bases, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic amino acids, and salts with acidic amino acids. Suitable examples of salts with inorganic bases include alkali metal salts such as sodium salts and potassium salts; alkaline earth metal salts such as calcium salts and magnesium salts; aluminum salts; and ammonium salts. Suitable examples of salts with organic bases include salts with trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine, tromethamine [tris(hydroxymethyl)methylamine], tert-butylamine, cyclohexylamine, benzylamine, dicyclohexylamine, and N,N-dibenzylethylenediamine. Suitable examples of salts with inorganic acids include salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, and phosphoric acid. Suitable examples of salts with organic acids include salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid. Suitable examples of salts with basic amino acids include salts with arginine, lysine, and ornithine. Suitable examples of salts with acidic amino acids include salts with aspartic acid and glutamic acid.

[0042] If compound (I) is obtained as a free compound, it can be converted to the desired salt by a method known to the present day. Alternatively, if compound (I) is obtained as a salt, it can be converted to a free form or another type of salt of the present day by a method known to the present day.

[0043] In this specification, "neurodegeneration" refers to the occurrence of one or more abnormalities among neurite regression, neurite fragmentation, neurite disappearance, cell body atrophy, cell body fragmentation, and cell body disappearance. Generally, neurodegeneration can be detected and evaluated using cell death as an indicator (in practice, as the reciprocal of the number of living nerve cells).

[0044] Furthermore, in neurodegeneration caused by abnormal elongation of hexanucleotide (or trinucleotide) repeat sequences, neurodegeneration can be detected and evaluated using RNA foci, which consists of RNA transcribed from the repeat sequence, or DPRs produced by RAN translation from the RNA. Both RNA foci and DPRs can be detected and quantified by well-known and conventional methods. RNA foci may be detected, for example, by analyzing nerve cells using the FISH (fluorescence in situ hybridization) method with an oligonucleotide (preferably labeled with fluorescence, etc.) containing a sequence complementary to the repeat sequence as a probe, and detecting the fluorescent signal dots (for example, dots with a diameter of 0.60 μm or more) present in the nucleus of the cells. The RNA foci generation level may then be quantitatively evaluated by measuring the number of dots per nerve cell or per unit area (for example, the area of ​​the observation field, the area of ​​the nuclear staining area in the observation field, etc.). DPRs may be detected and quantified, for example, by using an antibody against the DPRs that may arise from the hexa(or tri)nucleotide repeat sequence in a lysate or extract derived from nerve cells, using a well-known and conventional immunological analysis technique (e.g., ELISA). Alternatively, nerve cells may be detected and quantified by immunostaining using the antibody, and the DPRs identified as antibody-positive dots.

[0045] In one embodiment, the rate of suppression of cytopathic changes in motor neurons by a certain compound (test compound) may be calculated using the following formula, using motor neurons derived from iPS cells of ALS patients. Inhibitory activity of the test compound in motor neuron degeneration = ((XC) / (TC)) × 100 X: Number of motor neurons in the test compound group y days after the start of culture, C: Number of motor neurons in the DMSO group y days after the start of culture, T: Number of motor neurons x days after the start of culture Here, x is selected as any day before spontaneous cell death occurs in the subject, and y is selected as any day during which spontaneous cell death occurs in the subject.

[0046] Compound (I) is excellent at suppressing the generation of RNA foci and DPRs caused by abnormal elongation of G4C2 repeats; therefore, the agent of the present invention can be suitably used as a preventive or therapeutic agent for neurodegenerative diseases accompanied by abnormal elongation of the aforementioned repeats. Furthermore, since it is believed that there is a common mechanism independent of sequence in the process by which RNA foci and DPRs are generated from hexa(or tri)nucleotide repeats, the agent of the present invention is expected to be a preventive or therapeutic agent for neurodegenerative diseases in general accompanied by abnormal elongation of hexa(or tri)nucleotides. The aforementioned neurodegenerative diseases include motor neuron diseases and dementia. Examples of the aforementioned motor neuron diseases include amyotrophic lateral sclerosis (ALS), progressive bulbar palsy, progressive muscular atrophy, primary lateral sclerosis, progressive pseudobulbar palsy, spinal muscular atrophy, Parkinson's disease, multiple system atrophy, Huntington's disease, spinocerebellar degeneration, myotonic dystrophy, fragile X syndrome-related disorders, oculopharyngeal myopathy, Fuchs corneal dystrophy, and spinal and bulbar muscular atrophy. In this specification, these diseases may be referred to as "motor neuron diseases." Of these, amyotrophic lateral sclerosis, spinal and bulbar muscular atrophy, myotonic dystrophy, Parkinson's disease, Huntington's disease, fragile X syndrome-related disorders, and spinocerebellar degeneration are particularly preferred examples of target motor neuron diseases for the agent according to the present invention. Examples of dementia include frontotemporal dementia (FTD) and Lewy body dementia, with FTD being a particularly suitable target dementia disease. The aforementioned preventive or therapeutic agent can be used for the aforementioned disease, regardless of whether it is sporadic or familial. The aforementioned preventive or therapeutic agent can be used as a preventive or therapeutic agent for the aforementioned disease in mammals (e.g., mice, rats, hamsters, rabbits, cats, dogs, cattle, sheep, monkeys, humans, etc.). The agent of the present invention may contain one compound (I), or it may contain two or more compounds in combination.

[0047] Compound (I) exhibits excellent pharmacokinetics (e.g., drug half-life in blood, brain penetration, metabolic stability) and low toxicity (e.g., superior as a pharmaceutical in terms of acute toxicity, chronic toxicity, genotoxicity, reproductive toxicity, cardiotoxicity, drug interactions, carcinogenicity, etc.), and can be safely administered orally or parenterally to mammals (e.g., humans, monkeys, cattle, horses, pigs, mice, rats, hamsters, rabbits, cats, dogs, sheep, goats) as a pharmaceutical in itself or as a pharmaceutical composition mixed with pharmaceutically acceptable carriers, etc. Parenteral administration includes intravenous, intramuscular, subcutaneous, intraorgan, intranasal, intradermal, ophthalmic, intracerebral, intrarectal, intravaginal, intraperitoneal, intratumoral, proximal to tumor, and direct administration to the lesion.

[0048] The dosage of compound (I) varies depending on the route of administration and symptoms, but for example, when administered orally to a patient with amyotrophic lateral sclerosis (adult, weighing 40-80 kg, e.g., 60 kg), the dosage is, for example, 0.001-1000 mg / kg body weight per day, preferably 0.01-100 mg / kg body weight per day, and more preferably 0.1-10 mg / kg body weight per day. This amount can be administered in 1 to 3 divided doses per day.

[0049] The agent of the present invention can be used by compound (I) alone or as a pharmaceutical composition of compound (I) mixed with a pharmaceutically acceptable carrier, according to methods known as pharmaceutical manufacturing methods (e.g., methods described in the Japanese Pharmacopoeia). The agent (pharmaceutical, pharmaceutical composition) of the present invention can be used, for example, as tablets (including sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets, buccal tablets, etc.), pills, powders, granules, capsules (including soft capsules, microcapsules), lozenges, syrups, liquids, emulsions, suspensions, controlled-release formulations (e.g., immediate-release formulations, sustained-release formulations, sustained-release microcapsules), aerosols, and films (e.g., orally disintegrating films). It can be safely administered orally or parenterally (e.g., intravenously, intramuscularly, subcutaneously, intra-organally, intranasally, intradermally, ophthalmally, intracerebrally, intrarectally, vaginally, intraperitoneally) as oral mucosal adhesive film, injections (e.g., subcutaneous injections, intracutaneous injections, intramuscular injections, intramuscular injections, intraperitoneal injections), intrainfusions, transdermal formulations, ophthalmic formulations, etc.

[0050] The aforementioned "pharmaceutically acceptable carriers" include various organic or inorganic carriers commonly used as starting materials. For example, in solid formulations, excipients, lubricants, binders, and disintegrants are used, while in liquid formulations, solvents, solubilizers, suspending agents, isotonic agents, buffers, and analgesics are used. Additionally, preservatives, antioxidants, colorants, sweeteners, and other formulation additives may be used as needed. Examples of excipients include lactose, sucrose, D-mannitol, starch, corn starch, crystalline cellulose, and light anhydrous silicic acid. Examples of lubricants include magnesium stearate, calcium stearate, talc, and colloidal silica. Examples of binders include crystalline cellulose, sucrose, D-mannitol, dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, starch, sucrose, gelatin, methylcellulose, and sodium carboxymethylcellulose. Examples of disintegrants include starch, carboxymethylcellulose, carboxymethylcellulose calcium, carboxymethyl starch sodium, and L-hydroxypropylcellulose. Examples of solvents include sterile water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil, and olive oil. Examples of solubilizers include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, and sodium citrate. Examples of suspending agents include surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glyceryl monostearate; and hydrophilic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose. Examples of isotonic agents include glucose, D-sorbitol, sodium chloride, glycerin, and D-mannitol. Examples of buffering agents include buffer solutions such as phosphates, acetates, carbonates, and citrates. An example of an analgesic agent is benzyl alcohol. Examples of preservatives include para-hydroxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenylethyl alcohol, dehydroacetic acid, and sorbic acid. Examples of antioxidants include sulfites, ascorbic acid, and α-tocopherol.

[0051] Pharmaceutical compositions (formulations) vary depending on the dosage form, administration method, carrier, etc., but they can be manufactured by conventional methods by adding compound (I) in a ratio of typically 0.01 to 100% (w / w), preferably 0.1 to 95% (w / w), relative to the total amount of the formulation.

[0052] Compound (I) may be used alone as a drug (pharmaceutical, pharmaceutical composition) or in combination of two or more compounds. Unless otherwise specified herein, compound (I) also includes combinations of multiple compounds. Compound (I) may also be used in combination with other active ingredients (hereinafter abbreviated as "combination drugs").

[0053] Examples of concomitant medications include the following: Benzodiazepines (chlordiazepoxide, diazepam, potassium clorazepate, lorazepam, clonazepam, alprazolam, etc.), L-type calcium channel inhibitors (pregabalin, etc.), tricyclic or tetracyclic antidepressants (imipramine hydrochloride, amitriptyline hydrochloride, desipramine hydrochloride, clomipramine hydrochloride, etc.), selective serotonin reuptake inhibitors (fluvoxamine maleate, floxetine hydrochloride, citalopram bromide, sertraline hydrochloride) Paroxetine hydrochloride, escitalopram oxalate, etc.), serotonin-norepinephrine reuptake inhibitors (venlafaxine hydrochloride, duloxetine hydrochloride, desvenlafaxine hydrochloride, etc.), norepinephrine reuptake inhibitors (leboxetine mesylate, etc.), norepinephrine-dopamine reuptake inhibitors (bupropion hydrochloride, etc.), mirtazapine, trazodone hydrochloride, nefazodone hydrochloride, bupropion hydrochloride, cetiptiline maleate, 5-HT 1AAgonists (buspirone hydrochloride, tandospirone citrate, osemozotane hydrochloride, etc.), 5-HT3 antagonists (siamemazine, etc.), non-cardiac selective β-blockers (propranolol hydrochloride, oxyprenolol hydrochloride, etc.), histamine H1 antagonists (hydroxyzine hydrochloride, etc.), antischizophrenia medications (chlorpromazine, haloperidol, sulprid, clozapine, trifluoperazine hydrochloride, fluphenazine hydrochloride, olanzapine, quetiapine fumarate, risperidone, aripiprazole, etc.), CRF antagonists, other anxiolytics (meprobamate, etc.), tachykinin antagonists (MK-869, saledutant, etc.), drugs that act on metabotropic glutamate receptors Drugs, CCK antagonists, β3 adrenergic antagonists (amibegron hydrochloride, etc.), GAT-1 inhibitors (thiagabine hydrochloride, etc.), N-type calcium channel inhibitors, type 2 carbonic anhydrase inhibitors, NMDA glycine site agonists, NMDA antagonists (memantine, etc.), peripheral benzodiazepine receptor agonists, vasopressin antagonists, vasopressin V1b antagonists, vasopressin V1a antagonists, phosphodiesterase inhibitors, opioid antagonists, opioid agonists, uridine, nicotinic acid receptor agonists, thyroid hormones (T3, T4), TSH, TRH, MAO inhibitors (phenelzine sulfate, tranylcypromine sulfate, moclobemide, etc.), 5-HT 2A Antagonist, 5-HT 2AInverse agonists, COMT inhibitors (entacapone, etc.), bipolar disorder medications (lithium carbonate, sodium valproate, lamotrigine, riluzole, felbamate, etc.), cannabinoid CB1 antagonists (rimonabant, etc.), FAAH inhibitors, sodium channel blockers, anti-ADHD drugs (methylphenidate hydrochloride, methamphetamine hydrochloride, etc.), alcohol dependence medications, autism medications, chronic fatigue syndrome medications, convulsion medications, fibromyalgia medications, headache medications, insomnia medications (etizolam, zopiclone, triazolam, zolpidem, ramelteon, indipromone) (etc.), drugs for smoking cessation, myasthenia gravis, stroke, mania, hypersomnia, pain, mood disorders, autonomic nervous system disorders, male and female sexual dysfunction, migraine, pathological gambling, restless limb syndrome, substance addiction, alcohol-related disorders, irritable bowel syndrome, Alzheimer's disease (donepezil, galantamine, memantine, rivastigmine, etc.), Parkinson's disease (levodopa, carbidopa, benserazide, selegiline, zonisamide, endacapone, amantadine, taripe Xol, bramipexol, apomorphine, capelgoline, bromocriptine, istradefylline, trihexyphenidyl, promethazine, pergolide, etc.), Huntington's disease treatments (chlorpromazine hydrochloride, haloperidol, reserpine, etc.), Gaucher disease treatments (imiglucerase, taliglucerase alfa, veraglucerase alfa, eliglustat, miglustat, etc.), ALS treatments (riluzole, neurotrophic factors, etc.), multiple sclerosis treatments (fingolimod, interferon beta-1b, natalizumab, etc., molecular labeling Antiepileptic drugs (such as antiperspirants), anticonvulsants (phenytoin, carbamazepine, phenobarbital, primidone, zonizamide, sodium valproate, etosuximide, diazepam, nitrazepam, clonazepam, clobazam, gabapentin, topiramate, lamotrigine, levetiracetam, stiripentol, rufinamide, etc.), dyslipidemia drugs such as cholesterol-lowering drugs (statin series (pravastatin sodium, atrovastatin, simvastatin, rosuvastatin, etc.), fibrates (clofibrate, etc.), squalene synthesis inhibitors),Drugs for treating abnormal behavior or suppressing wandering tendencies due to dementia (sedatives, anxiolytics, etc.), apoptosis inhibitors, anti-obesity drugs, diabetes medications, hypertension medications, hypotension medications, rheumatoid arthritis medications (DMARDs), anticancer drugs, parathyroid drugs (PTH), calcium receptor antagonists, sex hormones or their derivatives (progesterone, estradiol, estradiol benzoate, etc.), neuronal differentiation promoters, nerve regeneration promoters, non-steroidal anti-inflammatory drugs (meloxicam, tenoxicam, indomethacin, ibuprofen, celecoxib, rofecoxib, aspirin, etc.), steroids (dexamethasone, cortisone acetate, etc.), anti-cytokine drugs (TNF inhibitors, MAP kinase inhibitors, etc.), antibody drugs, nucleic acids or nucleic acid derivatives, aptamer drugs, etc.

[0054] By combining compound (I) with the co-administered drug, (1) The dosage can be reduced compared to when compound (I) or the concomitant drug is administered alone. (2) Depending on the patient's symptoms (mild, severe, etc.), a drug can be selected to be used in combination with compound (I). (3) By selecting a co-administered drug with a different mechanism of action from compound (I), the treatment period can be extended. (4) By selecting a co-administered drug with a different mechanism of action from compound (I), the therapeutic effect can be sustained. (5) By using compound (I) in combination with the co-administered drug, excellent effects such as synergistic effects can be obtained.

[0055] Hereinafter, the use of compound (I) in combination with the co-administered drug will be referred to as "the co-administered agent of the present invention." When using the combination agent of the present invention, the timing of administration of compound (I) and the concomitant drug is not limited. Compound (I) or its pharmaceutical composition and the concomitant drug or its pharmaceutical composition may be administered simultaneously to the target patient, or they may be administered with a time difference. The dosage of the concomitant drug should be in accordance with clinically used dosages and can be appropriately selected depending on the target patient, route of administration, disease, combination, etc. The administration method of the combination agent of the present invention is not particularly limited, and it is sufficient that compound (I) and the combination drug are combined at the time of administration. Examples of such administration methods include: (1) Administration of a single formulation obtained by simultaneously formulating compound (I) and the co-administered drug. (2) Simultaneous administration of two formulations obtained by separately formulating compound (I) and the co-administered drug via the same route of administration. (3) Administration of two formulations obtained by separately formulating compound (I) and the co-administered drug, with a time difference between them, via the same route of administration. (4) Simultaneous administration of two formulations obtained by separately formulating compound (I) and the co-administered drug via different routes of administration. (5) Administration of two formulations obtained by separately formulating compound (I) and the co-administered drug, with a time difference between them, via different routes of administration (for example, administration in the order of compound (I); co-administered drug, or in the reverse order). These are some examples.

[0056] The combination agents of the present invention have low toxicity and can be safely administered orally or parenterally (e.g., topically, rectally, intravenously) by mixing compound (I) or / or the above combination agents with a pharmaceutically acceptable carrier according to known methods, for example, in the form of pharmaceutical compositions such as tablets (including sugar-coated tablets and film-coated tablets), powders, granules, capsules (including soft capsules), liquids, injections, suppositories, and sustained-release preparations. Injections can be administered intravenously, intramuscularly, subcutaneously, or intra-organally, or directly to the lesion. Examples of pharmaceutically acceptable carriers that may be used in the manufacture of the combination agent of the present invention include those described above.

[0057] The mixing ratio of compound (I) and the co-administered drug in the combination agent of the present invention can be appropriately selected depending on the target of administration, route of administration, disease, etc. For example, the content of compound (I) in the combination agent of the present invention varies depending on the form of the formulation, but is usually about 0.01 to 100% by weight, preferably about 0.1 to 50% by weight, and more preferably about 0.5 to 20% by weight relative to the total formulation. The content of the concomitant drug in the concomitant formulation of the present invention varies depending on the form of the formulation, but is usually about 0.01 to 100% by weight, preferably about 0.1 to 50% by weight, and more preferably about 0.5 to 20% by weight relative to the total formulation. [Examples]

[0058] The present invention will be further described in detail by the following examples and test examples, which are not intended to limit the present invention and may be modified without departing from the scope of the present invention. The test compounds used in the following test examples are as follows:

[0059] [Table 1] TIFF0007840054000010.tif53155

[0060] 4-(4-fluoro-2-methoxyphenyl)-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine (compound number 1) is described in Example 2 of WO 2012 / 160034. 1-(3-{[4-(2-methoxyphenyl)-1,3,5-triazine-2-yl]amino}phenyl)methanesulfonamide (compound number 2) is listed as Compound B1 in WO 2011 / 116951. 1-(3-{[4-(4-chloro-2-methoxyphenyl)-1,3,5-triazine-2-yl]amino}phenyl)methanesulfonamide (compound number 3) is listed as Compound B6 in WO 2011 / 116951. 1-[3-({4-[2-(benzyloxy)phenyl]-1,3,5-triazine-2-yl}amino)phenyl]methanesulfonamide (compound number 4b) and its trifluoroacetate (compound number 4a) are described as Compound B13 in WO 2011 / 116951. 4-{2-[(3,4-dichlorophenyl)methoxy]phenyl}-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine (compound number 5) is described in Example 61 of WO 2012 / 160034. Ethyl {[(3-{[4-(2,3-dihydro-1,4-benzodioxin-5-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ 6 Sulfanylidene carbamate (compound number 6) is described in Example 37 of WO 2012 / 160034. Ethyl{[(3-{[4-(2,3-dihydro-1-benzofuran-7-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ 6 Sulfanylidene carbamate (compound number 7) is described in Example 33 of WO 2012 / 160034.

[0061] Test Example 1 The effects of the compounds used in this invention on RNA foci were investigated using the ALS7 (C9orf72) cell line described in K. Imamura et al., Science Translational Medicine 2017, 9, eaaf3962. This cell line is a stable cell line created by introducing tetracycline-inducible Lhx3, Ngn2, and Isl1 genes into an iPS cell line established from a familial ALS patient with abnormal G4C2 repeat elongation of the C9orf72 gene. Therefore, ALS7 (C9orf72) cells rapidly differentiate into motor neurons (within approximately 7 days) when tetracycline or its derivatives are added to the culture medium, and after differentiation, they spontaneously undergo cell death (as described in the above-mentioned literature). Furthermore, the inventors found that the generation of RNA foci and DPRs occurs spontaneously prior to the aforementioned cell death. ALS7 (C9orf72) cells were cultured on feeder cells (mitomycin-treated SNL cells) using iPS cell maintenance medium consisting of Primate ES Cell medium (ReproCell, RCHEMD001A), 4 ng / ml hbFGF (Wako, 060-04543), 50 μg / ml G418 (Nacalai, 09380-86), and Penicillin-Streptomycin (Thermo Fisher Scientific, 15140-122). The method for seeding ALS7 (C9orf72) cells into assay plates is as follows: DMEM / F-12 (1:1) (Thermo Fisher Scientific, 11330-057), N2 supplement (Thermo Fisher Scientific, 17502-048), Penicillin-Streptomycin (Thermo Fisher Scientific, 15140-122), 10 ng / ml Recombinant human BDNF (PeproTech, 450-02), 10 ng / ml Recombinant human GDNF (PeproTech, 450-10), 10 ng / ml Recombinant human NT-3 (PeproTech, 450-03), 1 μM Retinoic acid (Sigma, R2625), 1 μg / ml Doxycycline (Clontech, 631311), 1 μM SAG (Enzo Life Sciences, ALX-270-426-M001), 10 μM Matrigel (BD Falcon, D2650) was diluted 20-fold with assay medium consisting of Y-27632 (Wako, 253-00513), and then coated with a 384-well plate (CellCarrier-384 Ultra, PerkinElmer, 6057300). Next, ALS7(C9orf72) cells were suspended in assay medium, 1 x 10⁶ cells per well. 4 The cells were seeded onto assay plates coated with Matrigel to form cells.

[0062] The method for fluorescent staining RNA foci in motor neurons differentiated from ALS7 (C9orf72) cells is as follows: ALS7 (C9orf72) cells seeded on assay plates according to the method described above were cultured four days after seeding with assay medium that did not contain Y-27632. Seven days after seeding, assay medium containing the test compound at a predetermined concentration (Retinoic Acid, Doxycycline, SAG, Y-27632-free) was added by changing the medium, and one day after adding the test compound, PFA (Wako, 163-20145) was added to fix the cells. After washing with PBS (Wako, 045-29795) three times, ice-cold methanol (Wako, 131-01826) was added, and the cells were allowed to stand at room temperature for 10 minutes. The DNA probe [5'-(Cy3)-CCCCGGCCCCGGCCCCGGCCCCGG-3' (SEQ ID NO: 1), SIGMA genosys, custom synthesized] was denatured at 80°C for 75 seconds, then prepared to a concentration of 2 μg / μL in a hybrid buffer consisting of 50% formamide (Wako, 066-02301), 2x SSC (Nippon Gene, 319-90015), 50 mM sodium phosphate (TEKNOVA, P2070), 10% Dextran sulfate (SIGMA, D8906-100G), 0.1 mg / mL yeast tRNA (invitrogen, 15401029), and RNase-free water (QIAGEN, 129112). This buffer was added to a plate and allowed to stand at 37°C for 16-24 hours to bind the probe to RNA foci. Wash buffer consisting of 50% formamide, 1x SSC, and RNase-free water was added and allowed to stand at 37°C for 30 minutes, followed by washing with PBS. Hoechst (invitrogen, H3569), diluted 5000-fold with PBS, was added and allowed to stand at room temperature for 20 minutes to stain the cell nuclei, followed by washing with PBS. RNA foci in the cell nucleus were detected by measuring the plates that had undergone the above treatment using a high-content analyzer (measuring the fluorescence of Cy3). The high-content analyzer used was the Opera Phenix from PerkinElmer. Figure 1 shows a typical image acquired with Opera.

[0063] The method for detecting the activity of the test compound is as follows: As a negative control, cells cultured in assay medium supplemented with DMSO instead of the test substance (motor neurons differentiated from ALS7 (C9orf72)) were used. Because cells sometimes overlapped and their nuclei overlapped, the area of ​​the nuclear staining region was used as an indicator of cell number. Changes in cell number were corrected by dividing the number of RNA foci in the nucleus by the area of ​​the nuclear staining region. The degree to which the test compound reduces the number of RNA foci in the negative control was defined as the RNA foci inhibitory activity of the test compound, and it was calculated using the following formula. RNA foci inhibitory activity of the test compound = 100 - X / Cx100 X: Number of RNA foci of the test compound group per unit nuclear area, C: Number of RNA foci of the DMSO group per unit nuclear area Table 2 below shows the activity values ​​of the test compound at concentrations of 1, 3, and 10 μmol / l.

[0064] [Table 2]

[0065] As shown in Table 2, treatment with any of the compounds effectively suppressed spontaneous RNA foci production in motor neurons derived from ALS7 (C9orf72) cells. In particular, compounds 1, 4a, 4b, and 5-7 suppressed RNA foci production by more than 40% over a wide concentration range of 1-10 μM. Therefore, it has been shown that the compound according to the present invention can effectively suppress RNA foci generation caused by abnormal elongation of G4C2 repeats.

[0066] Test Example 2 Next, the effect of the compounds used in this invention on RAN translation was investigated. The amount of poly-GP among the DPRs produced by RAN translation was measured using an electrochemiluminescence system (Meso Scale Diagnostics). ALS7 (C9orf72) cells were seeded into assay plates using the same method as in Test Example 1. Four days after seeding, assay medium without Y-27632 was added, and the cells were cultured until seven days after seeding. Then, assay medium containing the test compound at a predetermined concentration (Retinoic Acid, Doxycycline, SAG, Y-27632-free) was added by medium replacement. Two days after the addition of the test compound, the medium was removed, and the cells were lysed with Urea buffer consisting of 8M Urea (Wako, 219-00175), 4% CHAPS (Dojindo, 349-04722), and 30mM Tris-HCl (pH 8.0, Nippon Gene, 312-90061). Cell lysate was added to a MULTI-ARRAY 384 Well Plate (Meso Scale Diagnostics, L21XB-4), and the plate was agitated at room temperature and 700 rpm using a plate shaker (AS ONE, DM-301). The plate was then left to stand overnight at 4°C. After removing the cell lysate, a blocking buffer consisting of 5% Blocker A (Meso Scale Diagnostics, R93AA-1) and 1xTBS-T {TBS containing 0.05% Tween-20 (Bio-Rad, 170-6531) (Bio-Rad, 170-6435)} was added. The plate was agitated at room temperature and 700 rpm for 1 hour using a plate shaker, and then washed three times with a wash buffer consisting of 0.05% Tween-20 and PBS (Wako, 162-18547). After adding C9RANT Antibody (Novus Biologicals, NBP2-25018) diluted 10,000-fold with 1xTBS-T containing 1% Blocker A, the plate was agitated in a plate shaker at 700 rpm at 4°C for 2 hours, and then allowed to stand overnight at 4°C. After washing three times with wash buffer, SULFO-TAG Goat Anti-Rabbit Antibody (Meso Scale Diagnostics, R32AB-1) diluted 500-fold with 1xTBS-T containing 1% Blocker A was added, and the plate was agitated in a plate shaker at 700 rpm at room temperature for 2 hours.After washing three times with Wash buffer, 2xMSD Read Buffer T with Surfactant (Meso Scale Diagnostics, R92TC-1) was added, and the signal originating from the C9RANT Antibody was measured using MESO SECTOR S600 (Meso Scale Diagnostics).

[0067] The method for detecting the activity of the test compound is as follows: For negative controls, cells cultured in assay medium supplemented with DMSO instead of the test substance were used. The degree to which the test compound reduces the amount of poly-GP in the negative control was defined as the RAN translation inhibitory activity of the test compound, and it was calculated using the following formula. RAN translation inhibitory activity of the test compound = 100 - X / C × 100 X: Poly-GP amount in the test compound group, C: Poly-GP amount in the DMSO group Table 3 below shows the activity values ​​of the test compound at concentrations of 1, 3, and 10 μmol / l.

[0068] [Table 3]

[0069] As shown in Table 3, poly-GP production in ALS7 (C9orf72) cell-derived motor neurons was suppressed at 10 μM in all treatments with the compound. Therefore, it has been shown that the compound according to the present invention can effectively suppress RAN translation caused by abnormal elongation of G4C2 repeats.

[0070] Test Example 3 To examine the cytotoxicity of the test compound, intracellular ATP levels were measured. ALS7 (C9orf72) cells were seeded into assay plates using the same method as in Test Example 1. Four days after seeding, assay medium without Y-27632 was added, and the cells were cultured until seven days after seeding. Then, assay medium containing the test compound at a predetermined concentration (Retinoic Acid, Doxycycline, SAG, Y-27632-free) was added by medium exchange, and the medium was removed 24 or 48 hours after the addition of the test compound, and the CellTiter-Glo Luminescent Cell Viability Assay (Promega, G7570) was performed.

[0071] The method for detecting the cytotoxicity of the test compound is as follows: For negative controls, cells cultured in assay medium supplemented with DMSO instead of the test substance were used. The degree of effect on the intracellular ATP amount of cells treated with the test compound was defined as the ATP amount of the test compound compared to the negative control, and was calculated using the following formula. ATP content of the test compound compared to the negative control = X / C × 100 X: ATP amount of the test compound group, C: ATP amount of the DMSO group Cytotoxicity was determined if the intracellular ATP level was less than 0.5 times or greater than 1.5 times that of the negative control. Table 4 below shows the intracellular ATP levels at test compound concentrations of 1, 3, and 10 μmol / l.

[0072] [Table 4]

[0073] As shown in Table 4, in all cases of treatment with any of the compounds, the intracellular ATP levels in ALS7 (C9orf72) cell-derived motor neurons were within the range of 0.5 to 1.5 times the intracellular ATP levels of the negative control. Since the levels remained within this range even after 48 hours of treatment with each compound, the cytotoxicity of the compounds according to the present invention is considered to be sufficiently low. In particular, with compounds 1, 6, and 7, the intracellular ATP levels were maintained at 80% or more of the negative control even after 48 hours of treatment at concentrations in the 1-10 μM range, indicating that their cytotoxicity is considered to be very low. These results demonstrate that the compounds according to the present invention can effectively suppress RNA foci and RAN translation caused by abnormal G4C2 repeat elongation within a concentration range with sufficiently low cytotoxicity.

[0074] The compounds shown in Table 5 below, Reference Examples 1-30, are also expected to suppress RNA foci and RAN translation, similar to compounds 1-7.

[0075] [Table 5-1]

[0076] [Table 5-2]

[0077] [Table 5-3]

[0078] [Table 5-4]

[0079] [Table 5-5]

[0080] Examples of formulations A pharmaceutical product containing the compound of the present invention as an active ingredient can be manufactured, for example, by the following formulation. 1. Capsules (1) 10 mg of the compound obtained in Example 1 (2) Lactose 90mg (3) Microcrystalline cellulose 70 mg (4) Magnesium stearate 10 mg 1 capsule contains 180 mg After mixing the entire amounts of (1), (2), and (3) with 5 mg of (4), granulate the mixture, add the remaining 5 mg of (4), and enclose the whole mixture in gelatin capsules.

[0081] 2. Tablets (1) 10 mg of the compound obtained in Example 1 (2) Lactose 35 mg (3) Corn starch 150 mg (4) Microcrystalline cellulose 30 mg (5) Magnesium stearate 5 mg 1 tablet 230mg After mixing the entire amounts of (1), (2), and (3) above with 20 mg of (4) and 2.5 mg of (5), the mixture is granulated. Then, 10 mg of the remaining (4) and 2.5 mg of (5) are added to the granules, and the mixture is compressed to form tablets. [Industrial applicability]

[0082] The compound according to the present invention can effectively suppress the generation of RNA foci and DPRs by RAN translation in neurodegenerative diseases (e.g., ALS or FTD) caused by abnormal elongation of G4C2 repeats, thereby potentially preventing or treating said diseases. Some neurodegenerative diseases are caused by abnormal elongation of repeats consisting of different hexa(or tri)nucleotide sequences, and in all cases, the generation of RNA foci and DPRs is considered to be the main cytotoxicity leading to neurodegeneration. Furthermore, the existence of a sequence-independent common mechanism has been suggested in the process of generating these RNA foci and DPRs. Therefore, the compounds according to the present invention are expected to contribute as preventive or therapeutic agents not only to neurodegenerative diseases caused by abnormal elongation of G4C2 repeats, but also to neurodegenerative diseases in general caused by abnormal elongation of nucleotide repeats.

[0083] This application is based on Japanese Patent Application No. 2020-058414, filed in Japan on March 27, 2020, the contents of which are fully incorporated herein. [Sequence List] TIFF0007840054000019.tif153158

Claims

1. Equation (I) 【Chemistry 1】 [In the formula, R １ is equation (a-1) or (a-2) 【Chemistry 2】 (In the formula, R １１ and R １２ These each represent a hydrogen atom. R １３ is a hydrogen atom or C １－６ It shows an alkoxy-carbonyl group, R １４ C １－６ (Indicates an alkyl group.) The group represented by R ２ These are equations (b-1) to (b-3) 【Transformation 3】 (In the formula, R ２１ represents a C １－６ alkyl group, R ２２ This indicates a halogen atom. n represents 1, or R ２１ C may be substituted with halogen atoms. ７－１６ It shows an aralkyl group, n represents 0. The group represented by m represents 0, L is C １－３ [Indicates an alkylene group.] An inhibitor of neuronal degeneration accompanied by abnormal extension of the G4C2 repeat, comprising a compound represented by or a salt thereof, A compound represented by formula (I), or a salt thereof, (1) 4-(4-fluoro-2-methoxyphenyl)-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (2) 1-[3-({4-[2-(benzyloxy)phenyl]-1,3,5-triazine-2-yl}amino)phenyl]methanesulfonamide, (3) 4-{2-[(3,4-dichlorophenyl)methoxy]phenyl}-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (4) Ethyl {[(3-{[4-(2,3-dihydro-1,4-benzodioxin-5-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ6-sulfanylidene}carbamate, (5) Ethyl {[(3-{[4-(2,3-dihydro-1-benzofuran-7-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ6-sulfanylidene}carbamate, and neuronal degeneration inhibitors selected from their salts.

2. Equation (I) 【Chemistry 4】 [In the formula, R １ is equation (a-1) or (a-2) 【Transformation 5】 (In the formula, R １１ and R １２ These each represent a hydrogen atom. R １３ is a hydrogen atom or C １－６ It shows an alkoxy-carbonyl group, R １４ C １－６ (Indicates an alkyl group.) The group represented by R ２ These are equations (b-1) to (b-3) 【Transformation 6】 (In the formula, R ２１ C １－６ It shows an alkyl group, R ２２ This indicates a halogen atom. n represents 1, or R ２１ C may be substituted with halogen atoms. ７－１６ It shows an aralkyl group, n represents 0. The group represented by m represents 0, L is C １－３ [Indicates an alkylene group.] A preventive or therapeutic agent for motor neuron disease or dementia accompanied by abnormal extension of the G4C2 repeat, comprising a compound represented by or a salt thereof, A compound represented by formula (I), or a salt thereof, (1) 4-(4-fluoro-2-methoxyphenyl)-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (2) 1-[3-({4-[2-(benzyloxy)phenyl]-1,3,5-triazine-2-yl}amino)phenyl]methanesulfonamide, (3) 4-{2-[(3,4-dichlorophenyl)methoxy]phenyl}-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (4) Ethyl {[(3-{[4-(2,3-dihydro-1,4-benzodioxin-5-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ6-sulfanylidene}carbamate, (5) Ethyl {[(3-{[4-(2,3-dihydro-1-benzofuran-7-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ6-sulfanylidene}carbamate, and preventive or therapeutic agents selected from those salts.

3. The preventive or therapeutic agent according to claim 2, wherein the motor neuron disease or dementia is a motor neuron disease or dementia accompanied by abnormal elongation of hexanucleotide repeats.

4. The preventive or therapeutic agent according to claim 2 or 3, wherein the motor neuron disease is amyotrophic lateral sclerosis.

5. The preventive or therapeutic agent according to claim 2 or 3, wherein the dementia is frontotemporal dementia.

6. For manufacturing agents for the prevention or treatment of motor neuron disease or dementia involving abnormal elongation of G4C2 repeats, Equation (I) 【Transformation 7】 [In the formula, R １ is equation (a-1) or (a-2) 【Transformation 8】 (In the formula, R １１ and R １２ These each represent a hydrogen atom. R １３ is a hydrogen atom or C １－６ It shows an alkoxy-carbonyl group, R １４ C １－６ (Indicates an alkyl group.) R ２ These are equations (b-1) to (b-3) 【Chemistry 9】 (In the formula, R ２１ C １－６ It shows an alkyl group, R ２２ This indicates a halogen atom. n represents 1, or R ２１ C may be substituted with halogen atoms. ７－１６ It shows an aralkyl group, n represents 0. The group represented by m represents 0, L is C １－３ [Indicates an alkylene group.] The use of a compound represented by or a salt thereof, A compound represented by formula (I), or a salt thereof, (1) 4-(4-fluoro-2-methoxyphenyl)-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (2) 1-[3-({4-[2-(benzyloxy)phenyl]-1,3,5-triazine-2-yl}amino)phenyl]methanesulfonamide, (3) 4-{2-[(3,4-dichlorophenyl)methoxy]phenyl}-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (4) Ethyl {[(3-{[4-(2,3-dihydro-1,4-benzodioxin-5-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ6-sulfanylidene}carbamate, (5) Ethyl {[(3-{[4-(2,3-dihydro-1-benzofuran-7-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ6-sulfanylidene}carbamate, and a selection of those salts for use.

7. Formula (I) for manufacturing a preventive or therapeutic agent for neurodegenerative diseases involving abnormal extension of the G4C2 repeat. 【Chemistry 10】 [In the formula, R １ is equation (a-1) or (a-2) 【Chemistry 11】 (In the formula, R １１ and R １２ These each represent a hydrogen atom. R １３ is a hydrogen atom or C １－６ It shows an alkoxy-carbonyl group, R １４ C １－６ (Indicates an alkyl group.) The group represented by R ２ These are equations (b-1) to (b-3) 【Chemistry 12】 (In the formula, R ２１ C １－６ It shows an alkyl group, R ２２ This indicates a halogen atom. n represents 1, or R ２１ C may be substituted with halogen atoms. ７－１６ It shows an aralkyl group, n represents 0. The group represented by m represents 0, L is C １－３ [Indicates an alkylene group.] The use of a compound represented by or a salt thereof, A compound represented by formula (I), or a salt thereof, (1) 4-(4-fluoro-2-methoxyphenyl)-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (2) 1-[3-({4-[2-(benzyloxy)phenyl]-1,3,5-triazine-2-yl}amino)phenyl]methanesulfonamide, (3) 4-{2-[(3,4-dichlorophenyl)methoxy]phenyl}-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (4) Ethyl {[(3-{[4-(2,3-dihydro-1,4-benzodioxin-5-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ6-sulfanylidene}carbamate, (5) Ethyl {[(3-{[4-(2,3-dihydro-1-benzofuran-7-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ6-sulfanylidene}carbamate, and a selection of those salts for use.

8. Equation (I) 【Chemistry 13】 [In the formula, R １ is equation (a-1) or (a-2) 【Chemistry 14】 (In the formula, R １１ and R １２ These each represent a hydrogen atom. R １３ is a hydrogen atom or C １－６ It shows an alkoxy-carbonyl group, R １４ C １－６ (Indicates an alkyl group.) The group represented by R ２ These are equations (b-1) to (b-3) 【Chemistry 15】 (In the formula, R ２１ C １－６ It shows an alkyl group, R ２２ This indicates a halogen atom. n represents 1, or R ２１ C may be substituted with halogen atoms. ７－１６ It shows an aralkyl group, n represents 0. The group represented by m represents 0, L is C １－３ [Indicates an alkylene group.] A composition for use in the prevention or treatment of motor neuron disease or dementia accompanied by abnormal G4C2 repeat elongation, comprising a compound represented by or a salt thereof, and a pharmaceutically acceptable carrier, A compound represented by formula (I), or a salt thereof, (1) 4-(4-fluoro-2-methoxyphenyl)-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (2) 1-[3-({4-[2-(benzyloxy)phenyl]-1,3,5-triazine-2-yl}amino)phenyl]methanesulfonamide, (3) 4-{2-[(3,4-dichlorophenyl)methoxy]phenyl}-N-{3-[(S-methanesulfonimidoyl)methyl]phenyl}-1,3,5-triazine-2-amine, (4) Ethyl {[(3-{[4-(2,3-dihydro-1,4-benzodioxin-5-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ6-sulfanylidene}carbamate, (5) Ethyl {[(3-{[4-(2,3-dihydro-1-benzofuran-7-yl)-1,3,5-triazine-2-yl]aminophenyl)methyl](methyl)oxo-λ6-sulfanylidene}carbamate, and compositions selected from their salts.

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