GM1 gangliosidosis preventive or therapeutic agent, and composition for preventing or treating GM1 gangliosidosis

Abstract

To provide a novel prophylactic or therapeutic for GM1 gangliosidosis, and a novel composition for preventing or treating GM1 gangliosidosis.SOLUTION: Provided is a prophylactic or therapeutic for GM1 gangliosidosis, comprising, as an active ingredient, at least one selected from the group consisting of amodiaquine, acacetin, sulfamerazine, ungerine, amiodarone, sertindole, delcorine, perphenazine, althiazide, diethylstilbestrol, thiethylperazine, harmol, skimmianine, succinylsulfathiazole, phillalbine, canavanine, harmaline, trihexyphenidyl, fluoxetine, lovastatin, haloperidol, prenylamine lactate, bromperidol, convolvamine, and miglustat, and pharmaceutically acceptable salts thereof, or solvates thereof.SELECTED DRAWING: Figure 5

Description

[Technical Field]

[0001] The present invention relates to a preventive or therapeutic agent for GM1 gangliosidosis, and a composition for preventing or treating GM1 gangliosidosis. This application claims priority based on Japanese Patent Application No. 2017-254888, filed on December 28, 2017, the contents of which are incorporated herein by reference. [Background technology]

[0002] GM1 ganglioside is a type of glycolipid and a molecule involved in cell signaling, etc. GM1 gangliosidosis is known as a disease in which GM1 is involved.

[0003] GM1 gangliosidosis is a lysosomal storage disorder, a congenital metabolic disorder in which GM1 accumulates, particularly in the nervous system (brain), due to a deficiency or abnormality of the enzyme involved in the hydrolysis of GM1 ganglioside. GM1 gangliosidosis is classified into infantile, juvenile, and adult forms depending on the time of onset and clinical course. In particular, infantile forms show developmental delays by 3-6 months of age, and most patients develop severe neurological disorders such as decerebrate posturing by the time they are one year old, and usually die by the age of 3-4 years.

[0004] In the development of therapeutic agents for GM1 gangliosidosis, for example, Non-Patent Document 1 proposes Z-YVAD-FMK, a caspase inhibitor, as a preventive agent for GM1 gangliosidosis. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Special Publication No. 2016-533770 Summary of the Invention [Problem to be solved by the invention]

[0006] Currently, no therapeutic agent for GM1 gangliosidosis has been found, and the development of new therapeutic agents is desired.

[0007] Therefore, an object of the present invention is to provide a novel agent for preventing or treating GM1 gangliosidosis, and a composition for preventing or treating GM1 gangliosidosis. [Means for solving the problem]

[0008] The present inventors have discovered a compound that inhibits GM1 ganglioside (hereinafter also referred to as "GM1") through a screening system using neural stem cells differentiated from patient-derived iPS cells, and have completed the present invention. That is, the present invention is as follows. [1] Amodiaquine, Acacetin, Sulfamerazine, Ungerine, Amiodarone, Sertindole, Delcorine, Perphenazine, Althiazide, Diethylstilbestrol, Thiethylperazine A preventive or therapeutic agent for GM1 gangliosidosis, characterized by containing at least one active ingredient selected from the group consisting of Perazine, Harmol, Skimmianine, Succinylsulfathiazole, Fillalbin, Canavanine, Harmaline, Trihexyphenidyl, Fluoxetine, Lovastatin, Haloperidol, Prenylamine lactate, Bromperidol, Convolamine, and Miglustat, as well as pharmaceutically acceptable salts thereof, or solvates thereof. [2] A preventive or therapeutic agent for GM1 gangliosidosis, comprising a compound represented by the following general formula (1), a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient:

[0009] [ka]

[0010] [In general formula (1), R 1 and R 2 each independently represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, an aryl group, an aralkyl group, or a cycloalkyl group. X represents a single bond or a divalent linking group. n represents 0, 1, or 2. R3 represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group. 4 represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms. 5 represents a hydrogen atom, a hydroxyl group, an alkyl group having 1 to 5 carbon atoms, or an alkoxy group having 1 to 5 carbon atoms. 6 represents a hydrogen atom, a hydroxyl group, a cyano group, an alkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group.] [3] A preventive or therapeutic agent for GM1 gangliosidosis, comprising, as an active ingredient, a compound represented by the following general formula (2), a pharmaceutically acceptable salt thereof, or a solvate thereof:

[0011] [ka]

[0012] [In general formula (2), R 10 represents a single bond or a divalent linking group. 11 represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group. 12 and R 13 are each independently a hydrogen atom, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group. 10 and n 11 R each independently represents an integer of 0 to 4. 12 and R 13 When there are multiple of each, they may be the same or different. 14 represents a hydrogen atom, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, or an alkoxy group having 1 to 5 carbon atoms. n 12 represents an integer from 0 to 4. 14When there are multiple, they may be the same or different. [4] A preventive or therapeutic agent for GM1 gangliosidosis, comprising a compound represented by the following general formula (3), a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient:

[0013] [ka]

[0014] [In general formula (3), R 20 represents a hydrogen atom, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group. 21 and R 24 R each independently represents a single bond or a divalent linking group. 22 and R 23 each independently represents a hydrogen atom, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a haloalkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, or a hydroxyalkyl group having 1 to 5 carbon atoms.] [5] A preventive or therapeutic agent for GM1 gangliosidosis, comprising a compound represented by the following general formula (4), a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient:

[0015] [ka]

[0016] [In general formula (4), R 30 and R 31 each independently represents a hydrogen atom, a halogen atom, a hydroxyl group, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, or an aralkyl group. The double line consisting of a solid line and a dotted line represents a single bond or a double bond.] [6] A preventive or therapeutic agent for GM1 gangliosidosis, comprising, as an active ingredient, a compound represented by the following general formula (5), a pharmaceutically acceptable salt thereof, or a solvate thereof:

[0017] [ka]

[0018] [In general formula (5), R 40 represents a single bond or a divalent linking group. 41 ~R 43 each independently represents a hydrogen atom, a hydroxyl group, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, or an aralkyl group. [7] A composition for preventing or treating GM1 gangliosidosis, comprising the agent for preventing or treating GM1 gangliosidosis according to any one of [1] to [6] and a pharmaceutically acceptable carrier. [Effects of the Invention]

[0019] According to the present invention, a new agent for preventing or treating GM1 gangliosidosis can be provided. [Brief explanation of the drawings]

[0020] [Figure 1] FIG. 1 is a diagram comparing GM1 accumulation in neural stem cells derived from healthy individuals and GM1 patients. [Figure 2] FIG. 1 shows the state of GM1 accumulation when a GM1-suppressing compound is used. [Figure 3] FIG. 1 shows the state of GM1 accumulation when a GM1-suppressing compound is used. [Figure 4] FIG. 1 shows the effect of intraperitoneal administration of a GM1-suppressing compound on GM1 accumulation in brain slices from GM1 model mice. [Figure 5]FIG. 1 shows the results of comparing the amount of GM1 ganglioside in the brain of GM1 model mice when a GM1-inhibitory compound was administered intraperitoneally. [Figure 6] FIG. 1 shows the synthesis and degradation processes of gangliosides. [Figure 7] This shows the results of an analysis of the expression levels of each enzyme gene in neural stem cells obtained by differentiation from normal (201B7) and disease-derived (A138 #1-3) iPS cells to which amodiaquine or thiethylperanzine was added. DETAILED DESCRIPTION OF THE INVENTION

[0021] [Preventive or therapeutic agent for GM1 gangliosidosis] In one embodiment, the present invention provides a steroid drug, such as Amodiaquine, Acacetin, Sulfamerazine, Ungerine, Amiodarone, Sertindole, Delcorine, Perphenazine, Althiazide, Diethylstilbestrol, Thiethylperazin The present invention provides a preventive or therapeutic agent for GM1 gangliosidosis, which contains, as an active ingredient, at least one member selected from the group consisting of Thiethylperazine, Harmol, Skimmianine, Succinylsulfathiazole, Fillalbin, Canavanine, Harmaline, Trihexyphenidyl, Fluoxetine, Lovastatin, Haloperidol, Prenylamine lactate, Bromperidol, Convolamine, and Miglustat, as well as pharmaceutically acceptable salts thereof, or solvates thereof.

[0022] In one embodiment, the present invention provides a preventive or therapeutic agent for GM1 gangliosidosis, comprising, as an active ingredient, a compound represented by the following general formula (1), a pharmaceutically acceptable salt thereof, or a solvate thereof:

[0023] [ka]

[0024] [In general formula (1), R 1 and R 2each independently represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, an aryl group, an aralkyl group, or a cycloalkyl group. X represents a single bond or a divalent linking group. n represents 0, 1, or 2. R 3 represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group. 4 represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms. 5 represents a hydrogen atom, a hydroxyl group, an alkyl group having 1 to 5 carbon atoms, or an alkoxy group having 1 to 5 carbon atoms. 6 represents a hydrogen atom, a hydroxyl group, a cyano group, an alkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group.]

[0025] R 1 and R 2 Specific examples of the alkyl group having 1 to 5 carbon atoms in the formula (I) include a methyl group, an ethyl group, a propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a tert-butyl group, a pentyl group, an isopentyl group, and a neopentyl group. R 1 and R 2 The aryl group in the formula (I) is preferably one having 6 to 18 carbon atoms, more preferably one having 6 to 10 carbon atoms, and specifically, a phenyl group is particularly preferred. R 1 and R 2 The aralkyl group in R 1 and R 2 In the above, the aryl group is preferably bonded to the aryl group. R 1 and R 2 The cycloalkyl group in the formula (I) is preferably a group in which one hydrogen atom has been removed from a monocycloalkane having 3 to 8 carbon atoms, and specific examples include cyclopentane, cyclohexane, and cyclooctane.

[0026] Examples of the divalent linking group for X include an alkylene group, -O-, -C(=O)-, -NH-, -S-, -S(=O)2-, or a combination thereof.

[0027] R 3 In the above, the alkyl group, aralkyl group, aryl group, and cycloalkyl group having 1 to 5 carbon atoms are R 1 and R 2 Examples of the above-mentioned examples include: R 4 As the alkyl group having 1 to 5 carbon atoms in R 1 and R 2 Examples of the above-mentioned examples include:

[0028] R 5 As the alkyl group having 1 to 5 carbon atoms in R 1 and R 2 Examples of the above-mentioned examples include: R 5 As the alkoxy group having 1 to 5 carbon atoms, the R part of -OR is R 1 and R 2 Examples of the alkyl group include the same alkyl groups having 1 to 5 carbon atoms as those mentioned above.

[0029] R 6 In the formula, the alkyl group, aralkyl group, aryl group, and cycloalkyl group having 1 to 5 carbon atoms are R 1 and R 2 Examples of the above-mentioned examples include: R 6 As the alkoxy group having 1 to 5 carbon atoms in R 5 Examples of the above-mentioned examples include:

[0030] Among the above, X is preferably a single bond, n is preferably 1, and R 3 ~R 5 is preferably a hydrogen atom.

[0031] The preventive or therapeutic agent for GM1 gangliosidosis of this embodiment preferably contains, as an active ingredient, a compound represented by the following general formula (1-1), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0032] [ka]

[0033] [In general formula (1-1), R 1 and R 2 each independently represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, an aryl group, an aralkyl group, or a cycloalkyl group. X represents a single bond or a divalent linking group. R 6 represents a hydrogen atom, a hydroxyl group, a cyano group, an alkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group.]

[0034] Among the above, R 6 is preferably a hydroxyl group, and R 1 and R 2 As the alkyl group, an alkyl group having 1 to 5 carbon atoms is preferred.

[0035] It is particularly preferable that the preventive or therapeutic agent for GM1 gangliosidosis of this embodiment contains, as an active ingredient, a compound represented by the following formula (1-1-1), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0036] [ka]

[0037] In one embodiment, the present invention provides a preventive or therapeutic agent for GM1 gangliosidosis, comprising, as an active ingredient, a compound represented by the following general formula (2), a pharmaceutically acceptable salt thereof, or a solvate thereof:

[0038] [ka]

[0039] [In general formula (2), R 10 represents a single bond or a divalent linking group. 11represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group. 12 and R 13 are each independently a hydrogen atom, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group. 10 and n 11 R each independently represents an integer of 0 to 4. 12 and R 13 When there are multiple of each, they may be the same or different. 14 represents a hydrogen atom, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, or an alkoxy group having 1 to 5 carbon atoms. n 12 represents an integer from 0 to 4. 14 When there are multiple, they may be the same or different.

[0040] R 10 Examples of the divalent linking group in include the same groups as those described above for X. R 11 In the above, examples of the alkyl group having 1 to 5 carbon atoms, the alkoxy group having 1 to 5 carbon atoms, the aralkyl group, the aryl group, and the cycloalkyl group include R 5 Examples of the above-mentioned examples include: R 11 In the case of the hydroxyalkyl group having 1 to 5 carbon atoms, the alkyl group portion is R 1 and R 2 Examples of the above-mentioned examples include: R 12 and R 13 In the above, examples of the alkyl group having 1 to 5 carbon atoms, the alkoxy group having 1 to 5 carbon atoms, the hydroxyalkyl group having 1 to 5 carbon atoms, the aralkyl group, the aryl group, and the cycloalkyl group include R 11 Examples of the above-mentioned examples include: R 12and R 13 Examples of the halogen atom in the formula include a fluorine atom, a chlorine atom, and a bromine atom. R 12 and R 13 In the case of the thioalkyl group having 1 to 5 carbon atoms, the alkyl group portion is R 1 and R 2 Examples of the above-mentioned examples include: R 14 In the formula (I), examples of the halogen atom, the alkyl group having 1 to 5 carbon atoms, the thioalkyl group having 1 to 5 carbon atoms, and the alkoxy group having 1 to 5 carbon atoms include R 12 and R 13 Examples of the above-mentioned examples include:

[0041] Among the above, R 13 and R 14 is preferably a hydrogen atom, and n 10 is preferably 1.

[0042] The preventive or therapeutic agent for GM1 gangliosidosis of this embodiment preferably contains, as an active ingredient, a compound represented by the following general formula (2-1), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0043] [ka]

[0044] [In general formula (2-1), R 10 represents a single bond or a divalent linking group. 11 represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group. 12 represents a hydrogen atom, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group.]

[0045] Among the above, R10 As R, an alkylene group is preferred. 10 The alkylene group in the formula (I) is preferably a group in which one hydrogen atom has been removed from a linear or branched alkyl group having 1 to 5 carbon atoms. As the alkyl group having 1 to 5 carbon atoms, R 1 and R 2 Examples of the above-mentioned examples include:

[0046] It is more preferable that the preventive or therapeutic agent for GM1 gangliosidosis of this embodiment contains, as an active ingredient, a compound represented by the following general formula (2-1-1), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0047] [ka]

[0048] [In general formula (2-1-1), R 11 represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group. 12 represents a hydrogen atom, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group.]

[0049] Among the above, R 11 is preferably an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 1 to 5 carbon atoms. 12 is preferably a halogen atom or a thioalkyl group having 1 to 5 carbon atoms.

[0050] It is particularly preferable that the preventive or therapeutic agent for GM1 gangliosidosis of this embodiment contains, as an active ingredient, a compound represented by the following formula (2-1-1-1) or (2-1-1-2), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0051] [ka]

[0052] In one embodiment, the present invention provides a preventive or therapeutic agent for GM1 gangliosidosis, comprising, as an active ingredient, a compound represented by the following general formula (3), a pharmaceutically acceptable salt thereof, or a solvate thereof:

[0053] [ka]

[0054] [In general formula (3), R 20 represents a hydrogen atom, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, an aralkyl group, an aryl group, or a cycloalkyl group. 21 and R 24 R each independently represents a single bond or a divalent linking group. 22 and R 23 each independently represents a hydrogen atom, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a haloalkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, or a hydroxyalkyl group having 1 to 5 carbon atoms.]

[0055] R 20 In the above, examples of the halogen atom, the alkyl group having 1 to 5 carbon atoms, the thioalkyl group having 1 to 5 carbon atoms, the alkoxy group having 1 to 5 carbon atoms, the hydroxyalkyl group having 1 to 5 carbon atoms, the aralkyl group, the aryl group, and the cycloalkyl group include R 12 Examples of the above-mentioned examples include: R 21 and R 24 Examples of the divalent linking group in include the same groups as those described above for X. R 22 and R 23In the above, examples of the halogen atom, the alkyl group having 1 to 5 carbon atoms, the thioalkyl group having 1 to 5 carbon atoms, the alkoxy group having 1 to 5 carbon atoms, and the hydroxyalkyl group having 1 to 5 carbon atoms include R 12 Examples of the above-mentioned examples include: R 22 and R 23 In the formula, examples of the haloalkyl group having 1 to 5 carbon atoms include alkyl fluoride, alkyl chloride, and alkyl bromide. 1 and R 2 Examples of the alkyl groups include those in which at least one hydrogen atom has been replaced with a halogen atom.

[0056] Among the above, R 20 R is preferably an alkyl group having 1 to 5 carbon atoms or an aralkyl group. 20 The aralkyl group in the formula (I) is preferably an aralkyl group in which an alkylene group having 1 to 5 carbon atoms is bonded to a phenyl group. R 21 and R 24 is preferably a single bond or —O—. R 22 and R 23 is preferably a hydrogen atom or a trifluoromethyl group.

[0057] It is particularly preferable that the preventive or therapeutic agent for GM1 gangliosidosis of this embodiment contains, as an active ingredient, a compound represented by the following formula (3-1) or (3-2), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0058] [ka]

[0059] In one embodiment, the present invention provides a preventive or therapeutic agent for GM1 gangliosidosis, comprising, as an active ingredient, a compound represented by the following general formula (4), a pharmaceutically acceptable salt thereof, or a solvate thereof:

[0060] [ka]

[0061] [In general formula (4), R 30 and R 31 each independently represents a hydrogen atom, a halogen atom, a hydroxyl group, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, or an aralkyl group. The double line consisting of a solid line and a dotted line represents a single bond or a double bond.]

[0062] R 30 and R 31 In the above, examples of the halogen atom, the alkyl group having 1 to 5 carbon atoms, the thioalkyl group having 1 to 5 carbon atoms, the alkoxy group having 1 to 5 carbon atoms, the hydroxyalkyl group having 1 to 5 carbon atoms, and the aralkyl group include R 12 Examples of the above-mentioned examples include:

[0063] Among the above, R 30 and R 31 As the alkyl group, an alkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, and a hydroxyl group are preferred.

[0064] It is particularly preferable that the preventive or therapeutic agent for GM1 gangliosidosis of this embodiment contains, as an active ingredient, a compound represented by the following formula (4-1) or (4-2), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0065] [ka]

[0066] In one embodiment, the present invention provides a preventive or therapeutic agent for GM1 gangliosidosis, comprising, as an active ingredient, a compound represented by the following general formula (5), a pharmaceutically acceptable salt thereof, or a solvate thereof:

[0067] [ka]

[0068] [In general formula (5), R 40 represents a single bond or a divalent linking group. 41 ~R 43 each independently represents a hydrogen atom, a hydroxyl group, a halogen atom, an alkyl group having 1 to 5 carbon atoms, a thioalkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a hydroxyalkyl group having 1 to 5 carbon atoms, or an aralkyl group.

[0069] R 40 Examples of the divalent linking group in include the same groups as those described above for X. R 41 ~R 43 In the formula (I), examples of the halogen atom, the alkyl group having 1 to 5 carbon atoms, the thioalkyl group having 1 to 5 carbon atoms, the alkoxy group having 1 to 5 carbon atoms, the hydroxyalkyl group having 1 to 5 carbon atoms, and the aralkyl group include R 12 Examples of the above-mentioned examples include:

[0070] Among the above, R 40 As R, an alkylene group is preferred. 40 The alkylene group in is preferably a group in which one hydrogen atom has been removed from a linear or branched alkyl group having 1 to 5 carbon atoms. Examples of the linear or branched alkyl group having 1 to 5 carbon atoms include R 1 and R 2 Examples of the above-mentioned examples include: R 41 ~R 43 is preferably a hydrogen atom or a halogen atom.

[0071] It is particularly preferable that the preventive or therapeutic agent for GM1 gangliosidosis of this embodiment contains, as an active ingredient, a compound represented by the following formula (5-1) or (5-2), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0072] [ka]

[0073] As the preventive or therapeutic agent for GM1 gangliosidosis of the present invention, the above-mentioned compound may be used in the form of a free form or a pharmaceutically acceptable salt, or may be used in the form of a solvate of the free form or a solvate of the salt.

[0074] The salt is not particularly limited as long as it is a pharmaceutically acceptable salt, and examples thereof include hydrochloride, sulfate, hydrobromide, hydroiodide, phosphate, nitrate, benzoate, methanesulfonate, 2-hydroxyethanesulfonate, p-toluenesulfonate, acetate, propanoate, oxalate, malonate, succinate, glutarate, adipate, tartrate, maleate, fumarate, malate, mandelate, etc. The solvate is not particularly limited as long as it is a pharmaceutically acceptable solvate, and examples thereof include hydrates, organic solvents, etc.

[0075] [Composition for preventing or treating GM1 gangliosidosis] In one embodiment, the present invention provides a composition for preventing or treating GM1 gangliosidosis, comprising the above-mentioned agent for preventing or treating GM1 gangliosidosis and a pharmaceutically acceptable carrier.

[0076] The composition for preventing or treating GM1 gangliosidosis of this embodiment can be administered orally in the form of, for example, tablets, coated tablets, pills, powders, granules, capsules, liquids, suspensions, emulsions, etc., or parenterally in the form of injections, suppositories, topical skin preparations, etc.

[0077] As the pharmaceutically acceptable carrier, those usually used in the preparation of pharmaceutical compositions can be used without any particular limitation. More specifically, for example, binders such as gelatin, corn starch, tragacanth gum, gum arabic, etc.; excipients such as starch, crystalline cellulose, etc.; swelling agents such as alginic acid, etc.; solvents for injections such as water, ethanol, glycerin, etc.; adhesives such as rubber-based adhesives, silicone-based adhesives, etc. The pharmaceutically acceptable carriers can be used alone or in combination of two or more.

[0078] The composition for preventing or treating GM1 gangliosidosis of this embodiment may further contain additives, such as lubricants such as calcium stearate and magnesium stearate, sweeteners such as sucrose, lactose, saccharin, and maltitol, flavorings such as peppermint and rhizome oil, stabilizers such as benzyl alcohol and phenol, buffers such as phosphates and sodium acetate, solubilizers such as benzyl benzoate and benzyl alcohol, antioxidants, and preservatives. The additives can be used alone or in combination of two or more.

[0079] (Administration method) The method of administration of the preventive or therapeutic agent for GM1 gangliosidosis or the composition for preventing or treating GM1 gangliosidosis is not particularly limited and may be determined appropriately depending on the patient's symptoms, weight, age, sex, etc. For example, tablets, coated tablets, pills, powders, granules, capsules, liquids, suspensions, emulsions, etc. are administered orally. Injections are administered intravenously alone or mixed with common replacement fluids such as glucose and amino acids, and may also be administered intraarterially, intramuscularly, intradermally, subcutaneously, or intraperitoneally as needed. Suppositories are administered rectally. External skin preparations are applied, pasted, or sprayed onto the affected area.

[0080] (Dosage) The dosage of a preventive or therapeutic agent for GM1 gangliosidosis or a composition for preventing or treating GM1 gangliosidosis varies depending on the patient's symptoms, body weight, age, sex, etc., and cannot be determined in general. However, for oral administration, for example, 1 μg to 10 g of active ingredient may be administered per day, for example, 0.01 to 2000 mg per day. For injections, for example, 0.1 μg to 1 g of active ingredient may be administered per day, for example, 0.001 to 200 mg per day. For suppositories, for example, 1 μg to 10 g of active ingredient may be administered per day, for example, 0.01 to 2000 mg per day. For topical skin preparations, for example, 1 μg to 10 g of active ingredient may be administered per day, for example, 0.01 to 2000 mg per day.

[0081] [Other embodiments] In one embodiment, the present invention provides a compound such as Amodiaquine, Acacetin, Sulfamerazine, Ungerine, Amiodarone, Sertindole, Delcorine, Perphenazine, Althiazide, Diethylstilbestrol, Th for the prevention or treatment of GM1 gangliosidosis. Provided is a compound selected from the group consisting of iethylperazine, Harmol, Skimmianine, Succinylsulfathiazole, Fillalbin, Canavanine, Harmaline, Trihexyphenidyl, Fluoxetine, Lovastatin, Haloperidol, Prenylamine lactate, Bromperidol, Convolamine, Miglustat, and compounds represented by the general formulae (1) to (5), or a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0082] In one embodiment, the present invention provides a compound selected from the group consisting of Amodiaquine, Acacetin, Sulfamerazine, Ungerine, Amiodarone, Sertindole, Delcorine, Perphenazine, Althiazide, Diethylstilbestrol, Thiethylperazin The present invention provides a method for preventing or treating GM1 gangliosidosis, which comprises administering to a patient in need thereof an effective amount of a compound selected from the group consisting of Thiethylperazine, Harmol, Skimmianine, Succinylsulfathiazole, Fillalbin, Canavanine, Harmaline, Trihexyphenidyl, Fluoxetine, Lovastatin, Haloperidol, Prenylamine lactate, Bromperidol, Convolamine, Miglustat, and the compounds represented by the general formulae (1) to (5), or a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0083] In one embodiment, the present invention relates to a compound such as amodiaquine, acacetin, sulfamerazine, ungerine, amiodarone, sertindole, delcorine, perphenazine, althiazide, diethylstilbestrol, or the like for producing an agent for preventing or treating GM1 gangliosidosis, or a composition for preventing or treating GM1 gangliosidosis. The present invention provides use of a compound selected from the group consisting of Ethylstilbestrol, Thiethylperazine, Harmol, Skimmianine, Succinylsulfathiazole, Fillalbin, Canavanine, Harmaline, Trihexyphenidyl, Fluoxetine, Lovastatin, Haloperidol, Prenylamine lactate, Bromperidol, Convolamine, Miglustat, and the compounds represented by the general formulas (1) to (5), or a pharmaceutically acceptable salt thereof, or a solvate thereof. [Example]

[0084] The present invention will be explained in more detail below by way of experimental examples, but the present invention is not limited to these examples.

[0085] [Experimental Example 1] [Establishment of skin-derived fibroblasts] Fibroblasts were established from skin biopsy explants from patients with GM1 gangliosidosis and healthy controls under an ethical committee-approved protocol and with informed consent. Skin samples from patients and healthy volunteers were minced and cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS). After confirming the appearance of fibroblasts, the fibroblasts were expanded to introduce reprogramming genes.

[0086] [Establishment and maintenance of iPS cells] iPS cells were established from the established human fibroblasts using the method described in N. Fusaki, H. Ban, A. Nishiyama, K. Saeki, M. Hasegawa, Proc. Jpn. Acad. Ser., B. Phys. Biol. Eci., 85, 348 (2009). Specifically, 1 day before infection, 5 × 10 cells were cultured per well in a 6-well plate. 5 Human fibroblasts were seeded onto the cells and then infected with a Sendai virus (SeV) vector containing the Oct3 / 4, Sox2, K1f4, and c-Myc genes at a multiplicity of infection (MOI) of 3. The SeV vector containing the Oct3 / 4, Sox2, K1f4, and c-Myc genes was prepared according to the method described in N. Fusaki, H. Ban, A. Nishiyama, K. Saeki, and M. Hasegawa, Proc. Jpn. Acad. Ser., B. Phys. Biol. Eci., 85, 348 (2009). Seven days after infection, the infected fibroblasts were harvested using trypsin and plated at 5.4 × 10 cells per 60-mm dish. 4 cells, or 1-2 x 10 per 100 mm dish 5 The cells were seeded onto MMC-treated MEF feeder cells. The next day, the medium was replaced with human iPS cell medium, and the culture was continued for 30 days after infection, and colonies were collected.

[0087] 20% KNOCKOUT™ serum replacement (KSR, Invitrogen), 2 mM L-glutamine, 1 × 10 -4 M non-essential amino acids (NEAA, Sigma), 1 × 10 -4Human iPS cells were maintained on mitomycin C (MMC)-treated MEF feeder cells using a human iPS medium containing DMEM / F12 (Sigma) supplemented with 2-mercaptoethanol (Sigma) of M, 0.5% penicillin and streptomycin (Nacalai Tesque, Japan), and 5 ng / mL basic fibroblast growth factor (bFGF, Wako, Japan).

[0088] [Differentiation into neural stem cells (Neural Stem Cell: NSC)] One day before the start of differentiation induction, human iPS cells were seeded at a density of 2.5×10 5 ~3×10 5 cells / well on a 6-well plate coated with Geltrex (Thermo Fisher Scientific) and cultured in a human iPS medium. The next day (the start day of induction: Day 0), the human iPS medium was removed and replaced with PSC Neural Induction Medium (Thermo Fisher Scientific). Subsequently, the medium was changed with PSC Neural Induction Medium once every two days, and the cells were cultured until day 7 (Day 7) after the start of culture to induce NSC. The induced NSC was detached using StemPro Accutase Cell Dissociation Reagent (Thermo Fisher Scientific) except for the medium, suspended in Neural Expansion Medium (Thermo Fisher Scientific), and seeded onto a 100 mm cell culture dish coated with Geltrex. The cells at this point were designated as P0 NSC (NSC at passage 0), and thereafter, the NSC was appropriately expanded in this medium and used in the experiment.

[0089] [Experimental Example 2] [Visualization of GM1 accumulation using FITC-CTB] Neural stem cells induced from iPS cells derived from healthy individuals and patients with GM1 gangliosidosis were fixed with 4% paraformaldehyde, permeabilized with 0.1% TritonX-100 / PBS, and blocked with 1% BSA solution. Subsequently, multiple staining was performed using FITC-CTB (a reagent that specifically stains GM1 ganglioside) and anti-nestin antibody (a molecule specifically expressed in neural stem cells: a neural stem cell marker). The results are shown in Figure 1. Nestin staining showed strong staining in NSCs from both healthy individuals and GM patients, confirming their differentiation into neural stem cells (Figure 1, bottom).On the other hand, FITC-CTB staining was barely observed in healthy individuals, but strong staining was observed in GM1 patients (Figure 1, top). These results confirmed that GM1 accumulates in neural stem cells derived from GM1 patients.

[0090] [Experimental Example 3] <Evaluation of compounds through drug screening using an imaging cytometer> Screening for therapeutic agents for GM1 gangliosidosis was carried out as follows. After coating a 96-well plate with Geltrex, 5 × 10 neural stem cells induced from iPS cells derived from healthy individuals and GM1 gangliosidosis patients suspended in Neural Expansion Medium were added to the plate. 4 Cells were seeded at a density of 1000 / well. Compounds from the known drug library were then added to each well to a final concentration of 5 μM, and the cells were cultured for 72 hours. After fixation with 4% paraformaldehyde, permeabilization with 0.1% Triton X-100 / PBS, and blocking with 1% BSA solution, the cells were fluorescently stained using FITC-CTB (a reagent that specifically stains GM1 ganglioside), Hoechst 33342 (a reagent that stains cell nuclei), and CellMask (a reagent that stains cell membranes). The fluorescence intensity of each well was measured by acquiring images using an imaging cytometer (IN Cell Analyzer 6000), and the amount of GM1 ganglioside per cell was calculated using the IN Cell Developer Toolbox protocol. Table 1 shows the results of the compounds examined.

[0091] [Table 1]

[0092] Among the compounds examined, the staining images of neural stem cells after drug treatment in several compounds that were identified as GM1 inhibitory compounds are shown in Figures 2 and 3. In the identified compounds, it was confirmed that the fluorescence of CTB was decreased and the accumulation of GM1 was suppressed, as compared with the examples without the addition of the drug (lower panels in Figures 2b and 3e), (lower panels in Figures 2c and d, Figures 3f to g).

[0093] [Experimental Example 4] <Change in the amount of GM1 ganglioside in the brain of a GM1 gangliosideosis model mouse after administration of a GM1 inhibitory compound> GM1 gangliosidosis model mice (BKO mice: mice with complete deficiency of β-Galactosidase, β-Gal(- / -)) were intraperitoneally administered with Amodiaquine (40 mg / kg) and Thiethylperanzine (6 mg / kg) twice a day for 6 days from postnatal day P9 to P15. For positive controls, the same procedures were performed using PBS instead of the drug solution. For negative controls, the same procedures were performed using PBS instead of the drug solution and β-Gal(+ / -) mice. Then, the mouse brains were embedded in OCT compound and sliced at a thickness of 5 μm. After fixation with 4% paraformaldehyde, the slices were blocked with 1% BSA solution and fluorescently stained with Alexa Fluor488-CTB and Hoechst 33342. In addition, sphingolipids were fractionated and purified from mouse brains based on the extraction method of Svennerholm and Fredman and used as samples. These samples were analyzed by Agilent6460 Triple Quadrupole LC / MS to quantify GM1 in the brain. The results of the fluorescence staining are shown in Figure 4.

[0094] In the negative controls, almost no fluorescence of CTB was observed (upper panel of Figure 4a). On the other hand, in the positive controls, strong fluorescence of CTB was observed and accumulation of GM1 was confirmed (upper panel of Figure 4b). On the other hand, in all cases treated with the GM1 inhibitory compound, the fluorescence of CTB was strongly suppressed, and it was confirmed that the accumulation of GM1 was suppressed (upper panels of Figures 4c and d). The quantitative results of GM1 are shown in Figure 5.

[0095] An increase in the amount of GM1 ganglioside was confirmed in the positive controls compared to the negative controls. On the other hand, in all cases treated with the drug solution, it was confirmed that the amount of GM1 ganglioside was decreased compared to the positive controls.

[0096] [Experimental Example 5] [Effect of Adding GM1 Inhibitory Compound on Autophagy] The synthesis and degradation processes of gangliosides are shown in Figure 6. Gangliosides GM1, GM2, and GM3 are produced by the sequential addition of monosaccharides and N-acetylneuraminic acid. Ganglioside degradation is carried out in a stepwise manner by the action of glycosidases in lysosomes. (1) to (7) in Figure 6 indicate the enzymes that act in each reaction. Figure 7 shows that the expression of NEU1 and β-GLU increases upon treatment with candidate compounds. Neural stem cells differentiated from normal (201B7) and disease-derived (A138 #1-3) iPS cells were seeded into wells, and amodiaquine or thiethylperanzine was added to a concentration of 5 μM. The cells were cultured for 72 hours. After culture, RNA was isolated and the expression levels of each enzyme gene were analyzed. In Figure 7, - indicates a sample without addition, amo indicates a sample with addition of amodiaquine, and thie indicates a sample with addition of thiethylperanzine. These results suggest that the action of GM1 inhibitors promotes the degradation of gangliosides in lysosomes and activates autophagy. [Industrial Applicability]

[0097] According to the present invention, a new agent for preventing or treating GM1 gangliosidosis can be provided.

Claims

1. A preventive or therapeutic agent for GM1 gangliosidosis, characterized by containing thiethylperazine, a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient.

2. A composition for preventing or treating GM1 gangliosidosis, comprising the agent for preventing or treating GM1 gangliosidosis according to claim 1 and a pharmaceutically acceptable carrier.

Images