Prophylactic or therapeutic for GM1 gangliosidosis, 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 disease, and is an inborn error of metabolism in which GM1 accumulates, especially in the nervous system (brain), due to a deficiency or abnormality in the enzyme involved in the hydrolysis of GM1 ganglioside. GM1 gangliosidosis is classified into infantile, juvenile, and adult types depending on the time of onset and clinical course. In particular, infantile types show developmental delays within 3 to 6 months of age, and most patients show severe neurological disorders such as decerebrate posturing by the time they reach one year of age, and usually die by the age of 3 to 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] At present, no therapeutic agent has been found for GM1 gangliosidosis, and the development of a novel therapeutic agent 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 inventors discovered a compound that expresses GM1 ganglioside (hereinafter also referred to as "GM1") through a screening system using neural stem cells differentiated from patient-derived iPS cells, and 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, comprising as an active ingredient at least one 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 pharmaceutical 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 pharma- ceutically 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 R 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 R 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 a compound represented by the following general formula (2), a pharma- ceutically acceptable salt thereof, or a solvate thereof as an active ingredient:

[0011] [ka]

[0012] [In general formula (2), R 10 R represents a single bond or a divalent linking group. 11 R 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 each independently 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. 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 multiple are present, 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 pharma- ceutically acceptable salt thereof, or a solvate thereof as an active ingredient:

[0013] [ka]

[0014] [In general formula (3), R 20 R 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 pharma- ceutically 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 a compound represented by the following general formula (5), a pharma- ceutically acceptable salt thereof, or a solvate thereof as an active ingredient:

[0017] [ka]

[0018] [In general formula (5), R 40 R 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 pharma- ceutical acceptable carrier. Effect of the Invention

[0019] According to the present invention, a novel preventive or therapeutic agent for GM1 gangliosidosis can be provided. [Brief description of the drawings]

[0020] [Figure 1] FIG. 1 shows a comparison of GM1 accumulation in neural stem cells derived from healthy individuals and GM1 patients. [Diagram 2] FIG. 1 shows the state of GM1 accumulation when a GM1-suppressing compound was used. [Diagram 3] FIG. 1 shows the state of GM1 accumulation when a GM1-suppressing compound was used. [Figure 4] FIG. 1 shows the effect of intraperitoneal administration of a GM1-suppressing compound on GM1 accumulation in brain slices of GM1 model mice. [Diagram 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 differentiated from normal (201B7) and disease-derived (A138 #1-3) iPS cells after addition of amodiaquine or thiethylperanzine. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] [Preventive or therapeutic agent for GM1 gangliosidosis] In one embodiment, the present invention provides Amodiaquine, Acacetin, Sulfamerazine, Ungerine, Amiodarone, Sertindole, Delcorine, Perphenazine, Althiazide, Diethylstilbestrol, Thiethylperazin The present invention provides an agent for preventing or treating 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 pharma- ceutically acceptable salts thereof, or solvates thereof.

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

[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 R 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 R 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 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 is preferably a group in which one hydrogen atom has been removed from a monocycloalkane having 3 to 8 carbon atoms, and specific examples thereof include cyclopentane, cyclohexane, and cyclooctane.

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

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

[0028] R 5 As the alkyl group having 1 to 5 carbon atoms, R 1 and R 2 Examples of the above-mentioned examples include those mentioned above. R 5 In the case of the alkoxy group having 1 to 5 carbon atoms, the R portion of -OR is R 1 and R 2 Examples of the alkyl group include the alkyl groups having 1 to 5 carbon atoms described above.

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

[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 pharma- ceutical 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 preferable.

[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 pharma- ceutical acceptable salt thereof, or a solvate thereof.

[0036] [ka]

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

[0038] [ka]

[0039] [In general formula (2), R 10 R represents a single bond or a divalent linking group. 11R 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 each independently 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. 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 multiple are present, they may be the same or different.

[0040] R 10 Examples of the divalent linking group in include the same as those mentioned above for X. R 11 In the above, 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 are preferably R 5 Examples of the above-mentioned examples include those mentioned above. 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 those mentioned above. R 12 and R 13 In the above, 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 are preferably R 11 Examples of the above-mentioned examples include those mentioned above. R 12and R 13 Examples of the halogen atom in 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 those mentioned above. R 14 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 in R 12 and R 13 Examples of the above-mentioned examples include those mentioned above.

[0041] Among the above, R 13 and R 14 is preferably a hydrogen atom, 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 pharma- ceutical acceptable salt thereof, or a solvate thereof.

[0043] [ka]

[0044] [In general formula (2-1), R 10 R represents a single bond or a divalent linking group. 11 R 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 preferable. 10 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. 1 and R 2 Examples of the above-mentioned examples include those mentioned above.

[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 pharma- ceutical acceptable salt thereof, or a solvate thereof.

[0047] [ka]

[0048] [In general formula (2-1-1), R 11 R 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 R 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 pharma- ceutically acceptable salt thereof, or a solvate thereof.

[0051] [ka]

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

[0053] [ka]

[0054] [In general formula (3), R 20 R 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 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, the aralkyl group, the aryl group, and the cycloalkyl group include R 12 Examples of the above-mentioned examples include those mentioned above. R 21 and R 24 Examples of the divalent linking group in include the same as those mentioned above for X. R 22 and R 23In 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, and the hydroxyalkyl group having 1 to 5 carbon atoms include R 12 Examples of the above-mentioned examples include those mentioned above. R 22 and R 23 In the formula, examples of the haloalkyl group having 1 to 5 carbon atoms include an alkyl fluoride group, an alkyl chloride group, and an alkyl bromide group. 1 and R 2 In the above-mentioned alkyl groups, at least one hydrogen atom is 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 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 pharma- ceutically acceptable salt thereof, or a solvate thereof.

[0058] [ka]

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

[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 those mentioned above.

[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 preferable.

[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 pharma- ceutically 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 a compound represented by the following general formula (5), a pharma- ceutically acceptable salt thereof, or a solvate thereof as an active ingredient.

[0067] [ka]

[0068] [In general formula (5), R 40 R 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 as those mentioned above for X. R 41 ~R 43 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 in R 12 Examples of the above-mentioned examples include those mentioned above.

[0070] Among the above, R 40 As R, an alkylene group is preferable. 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 those mentioned above. 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 pharma- ceutically 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 pharma- ceutically 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 pharma- ceutically 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 pharma- ceutically acceptable solvate, and examples thereof include hydrate, organic solvent solvate, 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 pharma- ceutically 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 generally used in the preparation of pharmaceutical compositions can be used without any particular limitation. More specifically, for example, binders such as gelatin, cornstarch, tragacanth gum, gum arabic, etc.; excipients such as starch and crystalline cellulose, swelling agents such as alginic acid, injection solvents such as water, ethanol, glycerin, etc.; adhesives such as rubber-based adhesives and silicone-based adhesives, etc. can be mentioned. 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 the present 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 saffron 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 may 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 appropriately determined according to the symptoms, weight, age, sex, etc. of the patient. For example, tablets, coated tablets, pills, powders, granules, capsules, liquids, suspensions, emulsions, etc. are administered orally. Injections are administered intravenously alone or in combination with normal replacement fluids such as glucose and amino acids, and further administered intraarterially, intramuscularly, intradermally, subcutaneously, or intraperitoneally as necessary. Suppositories are administered rectally. Skin external preparations are applied, pasted, or sprayed on the affected area.

[0080] (Dosage) The dosage of the preventive or therapeutic agent for GM1 gangliosidosis or the composition for preventing or treating GM1 gangliosidosis varies depending on the symptoms, body weight, age, sex, etc. of the patient and cannot be determined in general, but in the case of oral administration, for example, 1 μg to 10 g per day, for example, 0.01 to 2000 mg per day of the active ingredient may be administered. In the case of an injection, for example, 0.1 μg to 1 g per day, for example, 0.001 to 200 mg per day of the active ingredient may be administered. In the case of a suppository, for example, 1 μg to 10 g per day, for example, 0.01 to 2000 mg per day of the active ingredient may be administered. In the case of an external preparation for skin, for example, 1 μg to 10 g per day, for example, 0.01 to 2000 mg per day of the active ingredient may be administered.

[0081] [Other embodiments] In one embodiment, the present invention provides a pharmaceutical composition comprising Amodiaquine, Acacetin, Sulfamerazine, Ungerine, Amiodarone, Sertindole, Delcorine, Perphenazine, Althiazide, Diethylstilbestrol, Th for the prevention or treatment of GM1 gangliosidosis. The present invention provides 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 the compounds represented by the general formulas (1) to (5), or a pharma- ceutically acceptable salt or solvate thereof.

[0082] In one embodiment, the present invention provides Amodiaquine, Acacetin, Sulfamerazine, Ungerine, Amiodarone, Sertindole, Delcorine, Perphenazine, Althiazide, Diethylstilbestrol, Thiethylperazin The present invention provides a method for preventing or treating GM1 gangliosidosis, comprising administering to a patient in need of such prevention or treatment 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 general formulas (1) to (5), a pharma- ceutically acceptable salt thereof, or a solvate thereof.

[0083] In one embodiment, the present invention relates to a prophylactic or therapeutic agent for GM1 gangliosidosis, or a composition for the prevention or treatment of GM1 gangliosidosis, comprising Amodiaquine, Acacetin, Sulfamerazine, Ungerine, Amiodarone, Sertindole, Delcorine, Perphenazine, Althiazide, Diethylstilbestrol, or the like. 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 general formulas (1) to (5), or a pharma- ceutically acceptable salt thereof, or a solvate thereof. EXAMPLES

[0084] The present invention will be described in more detail below with reference to 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 explants of skin biopsies from patients with GM1 gangliosidosis and healthy controls under informed consent, according to a protocol approved by the ethical committee. Skin samples from patients and healthy subjects were finely 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 by 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 5 Human fibroblasts were seeded on the cells, and then the cells were infected with a Sendai virus (SeV) vector containing Oct3 / 4, Sox2, K1f4, and c-Myc genes at a multiplicity of infection (MOI) of 3. The SeV vector containing Oct3 / 4, Sox2, K1f4, and c-Myc genes was prepared according to 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). Seven days after infection, the infected fibroblasts were harvested using trypsin and 5.4 × 10 cells were cultured 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 cell medium containing 2-mercaptoethanol (Sigma) of M, 0.5% penicillin and streptomycin (Nacalai Tesque, Japan), and basic fibroblast growth factor (bFGF, Wako, Japan) at 5 ng / mL in DMEM / F12 (Sigma).

[0088] [Differentiation into Neural Stem Cells (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 cell medium. The next day (the start day of induction: Day 0), the human iPS cell 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 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 time were designated as P0 NSC (NSC at passage 0), and thereafter, 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 subjects and patients with GM1 gangliosidosis were fixed with 4% paraformaldehyde, permeabilized with 0.1% TritonX-100 / PBS, and blocked with 1% BSA solution. Then, 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 marker for neural stem cells). The results are shown in Figure 1. Nestin staining was strong in both NSCs from healthy subjects and GM patients, confirming their differentiation into neural stem cells (Fig. 1, bottom).On the other hand, FITC-CTB staining was barely observed in healthy subjects, but strong staining was observed in GM1 patients (Fig. 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 subjects and GM1 gangliosidosis patients suspended in Neural Expansion Medium were added to the plate. 4 The 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 5μM, and the cells were cultured for 72 hours. The cells were then fixed with 4% paraformaldehyde, permeabilized with 0.1% TritonX-100 / PBS, blocked with 1% BSA solution, and fluorescently stained using FITC-CTB (a reagent that specifically stains GM1 ganglioside), Hoechst 33342 (a reagent that stains the cell nucleus), and CellMask (a reagent that stains the cell membrane). 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 stained images of neural stem cells after drug treatment in several compounds that were hit as GM1 inhibitory compounds are shown in Figures 2 and 3. In the hit compounds, it was confirmed that the fluorescence of CTB was decreased and the accumulation of GM1 was suppressed as compared with the examples without addition of the drug (lower panels in Figures 2b and 3e), (lower panels in Figures 2c and d, and 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 Amodiaquine (40 mg / kg) and Thiethylperanzine (6 mg / kg) twice a day for 6 days from P9 to P15 after birth. For positive controls, PBS was used as the drug solution and the same procedure was performed. For negative controls, PBS was used as the drug solution and β-Gal(+ / -) mice were used, and the same procedure was performed. Then, the mouse brains were embedded in OCT compound and sliced at a thickness of 5 μm. After the slices were fixed with 4% paraformaldehyde, they 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 these samples were analyzed by Agilent6460 Triple Quadrupole LC / MS to quantify GM1 in the brain. The results of the fluorescent 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 the 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 process of gangliosides is 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 stepwise by the action of glycosidases in lysosomes. (1) to (7) in Figure 6 indicate the enzymes that act in each reaction. FIG. 7 shows that the expression of NEU1 and β-GLU is increased by treatment with the candidate compound. Neural stem cells obtained by differentiation 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, followed by culture for 72 hours. After culture, RNA was isolated and the expression level of each enzyme gene was analyzed. In FIG. 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 the GM1 inhibitory compound promotes the degradation of gangliosides in lysosomes and activates autophagy. [Industrial Applicability]

[0097] According to the present invention, a novel preventive or therapeutic agent for GM1 gangliosidosis can be provided.

Claims

1. A preventive or therapeutic agent for GM1 gangliosidosis, comprising thiethylperazine, a pharma- ceutical acceptable salt thereof, or a solvate thereof as an active ingredient.

2. A composition for preventing or treating GM1 gangliosidosis, comprising the GM1 gangliosidosis preventive or therapeutic agent according to claim 1 and a pharma- ceutical acceptable carrier.

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