Therapeutic agent for frontotemporal lobar degeneration and therapeutic composition

Abstract

The present invention addresses the problem of providing a therapeutic agent for frontotemporal lobe degenerative disease (FTLD), a composition for treating FTLD, and a method for treating FTLD, which have not been developed so far. The inventor of the present invention finds that: a prefrontal lobe cerebral cortex neuron is differentiated and induced by an iPS cell derived from an FTLD patient; a therapeutic agent for frontotemporal lobe degenerative disease (FTLD), comprising a compound represented by formula (1-1), a compound represented by formula (2-1), or a compound represented by formula (3-1), a pharmaceutically acceptable salt thereof, or a solvate thereof, is administered to a prefrontal cerebral cortex neuron exhibiting a pathological state of FTLD. Therefore, the pathological state of FTLD of the forehead lobe type cerebral cortical neurons is improved.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480066101.0 (22) Application Date 2024.11.01 (30) Priority Data 2023-188497 2023.11.02 JP 2024-173803 2024.10.02 JP (85) PCT International Application Entering National Phase Date 2026.04.15 (86) PCT International Application Application Data PCT / JP2024 / 039021 2024.11.01 (87) PCT International Application Publication Data WO2025 / 095099 JA 2025.05.08 (71) Applicant: Kai Pharmaceutical Co., Ltd. Address: Japan (72) Inventor: Hiroshi Kokubun (74) Patent Agency: Beijing Linda Liu Intellectual Property Agency (General Partnership) 11277 Patent Attorneys: Li Maojia, Li Enhua (51) Int.Cl. A61K 31 / 4045 (2006.01) A61K 31 / 42 (2006.01) A61K 31 / 4184 (2006.01) A61P 25 / 00 (2006.01) A61P 25 / 28 (2006.01) (54) Invention Title: Treatment Agent and Composition for Frontotemporal Degenerative Disease (57) Abstract: The object of the present invention is to provide a treatment agent or composition for frontotemporal degenerative disease (FTLD) for which no treatment has been developed to date, and a treatment method for FTLD. The inventors of this invention have discovered that by inducing prefrontal cortical neurons through differentiation of iPS cells derived from FTLD patients, and by administering a frontotemporal degenerative disease (FTLD) treatment agent comprising a compound of formula (1-1), a compound of formula (2-1), or a compound of formula (3-1), a pharmaceutically acceptable salt thereof, or a solvate thereof to prefrontal cortical neurons exhibiting the pathological state of FTLD, the pathological state of FTLD in prefrontal cortical neurons is improved.Claims 4 pages, Description 18 pages, Drawings 7 pages, CN 122028915 A 2026.05.12 CN 1 22 02 89 15 A 1. A therapeutic agent for frontotemporal lobe degenerative disease (FTLD), comprising: a compound represented by the following formula (1-1): In formula (1-1), R1 independently represents an alkyl group or a 4-hydroxyphenylethyl group having 1 to 6 carbon atoms, and n represents an integer from 1 to 3; a compound represented by the following formula (2-1): In formula (2-1), R21 is selected from the group consisting of hydrogen, a fatty acid acyl group having 2 to 18 carbon atoms, and an aromatic carboxylic acid acyl group having 7 to 9 carbon atoms; a compound represented by the following formula (3): In formula (3-1), R31 is selected from the group consisting of hydrogen, halogen, an alkyl group having 1 to 6 carbon atoms, and CF3, R32 is selected from the group consisting of alkoxy groups having 1 to 6 carbon atoms substituted with an imidazole group, optionally substituted nitrogen-containing aromatic heterocycles; R33 is selected from the group consisting of hydrogen, alkyl groups having 1 to 6 carbon atoms, and cycloalkyl groups having 3 to 6 carbon atoms; R34 is selected from the group consisting of carboxyl groups, cyano groups, and 1H-tetrazole groups; pharmaceutically acceptable salts of these or their solvates. 2. The FTLD therapeutic agent according to claim 1, wherein FTLD is one or more symptoms selected from behavioral variant frontotemporal dementia (bvFTD), semantic dementia (SD), and progressive nonfluent aphasia (PNFA). 3. The FTLD therapeutic agent according to claim 1 or 2, wherein n is 2 in formula (1-1). 4. The FTLD therapeutic agent according to claim 1 or 2, wherein R1 is n-propyl in formula (1-1). Claims 1 / 4 page 2 CN 122028915 A 5. The FTLD therapeutic agent according to claim 3, wherein R1 in formula (1-1) is n-propyl. 6. The FTLD therapeutic agent according to claim 1 or 2, wherein the compound represented by formula (1-1) is the compound represented by formula (1-2): namely 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one. 7. The FTLD therapeutic agent according to claim 1 or 2, wherein a pharmaceutically acceptable salt of the compound represented by formula (1-1) is the hydrochloride salt of the compound represented by formula (1-2): namely 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one hydrochloride. 8. The FTLD therapeutic agent according to claim 1 or 2, wherein the compound represented by formula (2-1) is the compound represented by formula (2-2): namely, 4-amino-N-(3,4-dimethyl-5-isoxazolyl)benzenesulfonamide.9. The FTLD therapeutic agent according to claim 1 or 2, wherein the compound represented by formula (3-1) is the compound represented by formula (3-2) below: Claims 2 / 4 pages 3 CN 122028915 A i.e. 4'-[[4-methyl-6-(1-methyl-1H-benzimidazol-2-yl)-2-propyl1H-benzimidazol-1-yl]methyl]biphenyl-2-carboxylic acid. 10. A pharmaceutical composition for the treatment of frontotemporal lobe degenerative disease (FTLD), comprising the following substances as active ingredients: A compound represented by the following formula (1-1): In formula (1-1), R1 independently represents an alkyl group or a 4-hydroxyphenylethyl group having 1 to 6 carbon atoms, and n represents an integer from 1 to 3; A compound represented by the following formula (2-1): In formula (2-1), R21 is selected from the group consisting of hydrogen, a fatty acid acyl group having 2 to 18 carbon atoms, and an aromatic carboxylic acid acyl group having 7 to 9 carbon atoms; A compound represented by the following formula (3-1): Claims 3 / 4 page 4 CN 122028915 A In formula (3-1), R31 is selected from the group consisting of hydrogen, halogen, an alkyl group having 1 to 6 carbon atoms, and CF3; R32 is selected from the group consisting of an alkoxy group having 1 to 6 carbon atoms substituted with an imidazolium group, and optionally a nitrogen-containing aromatic heterocycle that has been substituted. R33 is selected from the group consisting of hydrogen, alkyl groups having 1 to 6 carbon atoms, and cycloalkyl groups having 3 to 6 carbon atoms; R34 is selected from the group consisting of carboxyl groups, cyano groups, and 1H-tetrazole groups; pharmaceutically acceptable salts of these or their solvates. Claims 4 / 4 pages 5 CN 122028915 A Treatment Agents and Compositions for Frontotemporal Lobar Degeneration Technical Field

[0001] The object of the present invention is to develop treatment agents or compositions for frontotemporal lobar degeneration (FTLD). Background Art

[0002] Frontotemporal lobar degeneration (FTLD) is a type of dementia characterized by degeneration of the prefrontal and lateral temporal lobes, with symptoms mainly including behavioral and speech disorders. It is designated as a designated intractable disease in Japan (Designated Intractable Disease 127), with an estimated number of approximately 12,000 patients (Non-Patent Document 1), an estimated number of 50,000 to 60,000 patients in the United States, and an estimated number of more than 100,000 patients throughout Europe.

[0003] FTLD is broadly classified into three subtypes based on its clinical symptoms, with the characteristics shown in the following table:

[0004] • Behavioral variant frontotemporal dementia (bvFTD)

[0005] • Semantic dementia (SD)

[0006] • Progressive non-fluent aphasia (PNFA).

[0007] [Table 1]

[0008]

[0009] FTLD is a diverse disease in pathology and genetics. Based on related proteins, it is sometimes classified as: FTLD-tau (45%) with observed TAU accumulation, FTLD-TDP43 (45%) with observed TDP-43 accumulation, and FTLD-FUS (9%) with observed FUS accumulation. In addition, from a genetic perspective, GRN, MAPT, C9ORF72, and TARDBP genes are known to be representative mutated genes.

[0010] Frontotemporal dementia (FTD) refers to a disease group that comprehensively encompasses non-Alzheimer's type degenerative dementia diseases mainly caused by lesions of the motor cortex. The causes are known to involve TAU degeneration, TDP-43, and FUS. Diseases involving TDP-43 and FUS are divided into: diseases with motor impairment (amyotrophic lateral sclerosis) (FTLD-MND) and diseases without motor impairment (FTLD-nonMND). On the other hand, diseases caused by TAU degeneration are classified as: Pick's disease, which is mainly caused by 3R TAU degeneration, and the disease group mainly caused by 4R TAU degeneration (cortical basal ganglia degeneration, progressive supranuclear palsy, and argyrophilic granular dementia).

[0011] As a method for diagnosis based on pathological state, in the case of bvFTD, a diagnosis can be made by meeting at least 3 of the following evaluation items A to F.

[0012] A. Disinhibition behavior: Meeting any 1 or more of the following 3 symptoms.

[0013] 1) Social behavioral anomie specification 1 / 18 page 6 CN 122028915 A

[0014] 2) Lack of manners and etiquette

[0015] 3) Impulsive and reckless behavior

[0016] B. Lack of concern or energy

[0017] C. Lack of empathy and emotional involvement: Meeting any 2 or more of the following symptoms.

[0018] 1) Lack of responsiveness to the needs and emotions of others

[0019] 2) Decreased or lost interest in society, communication with others, or emotional resonance

[0020] D. Stereotyped / rigidity: Meeting any one or more of the following three symptoms.

[0021] 1) Repetition of simple actions

[0022] 2) Compulsive or ritualistic behaviors

[0023] 3) Stereotyped speech

[0024] E. Changes in oral preferences and eating habits: Meeting any one or more of the following three symptoms.

[0025] 1) Changes in dietary preferences

[0026] 2) Increased overeating, drinking, and smoking

[0027] 3) Oral exploration or pica

[0028] F. In neuropsychological examinations, although memory and visuospatial cognitive abilities are relatively maintained, executive dysfunction is observed.

[0029] In the case of SD, at least three of the following four were observed: A. Impairment in knowledge of the object (particularly significant impairment in low-frequency / low-contact speech), B. Surface alexia / agraphia, C. Repetition is maintained, D. Vocalization (grammar, spontaneous speech) is maintained.

[0030] Currently, treatment methods aimed at curing FTLD (improvement of cognitive function) have not been established, and symptomatic treatment is used for some symptoms. As such symptomatic treatments, for example, selective serotonin reuptake inhibitors (SSRIs) have been reported to be useful when administering antidepressants when behavioral disorders are obvious. In addition, although still at the case report level, antipsychotics and antiepileptics are known to be effective. However, these are not aimed at improving cognitive function, and it is hoped that drugs that can improve cognitive function will be developed.

[0031] Prior Art Documents

[0032] Non-Patent Documents

[0033] Non-Patent Document 1: Wada-Isoe K., et al., Epidemiological Survey of Frontotemporal Lobar Degeneration in Tottori Prefecture, Japan., Dement. Geriatr. Cogn. Dis. Extra. 2 (1), 381-386, 2012

[0034] Non-Patent Document 2: Imaizumi K., et al., Rostrocaudal Areal Patterning of Human PSC-Derived Cortical Neurons by FGF8 Signaling., eNeuro, 5 (2), 2018 Summary of the Invention

[0035] Problems to be Solved by the Invention

[0036] The object of the present invention is to provide a therapeutic agent or composition for frontotemporal lobe degeneration (FTLD) for which no therapeutic agent has been developed to date, or a treatment method for FTLD.

[0037] Solution to the Problem

[0038] The inventors of the present invention have discovered that by inducing prefrontal cortical neurons through differentiation from iPS cells derived from FTLD patients, and administering the following FTLD treatment agent to prefrontal cortical neurons exhibiting the pathological state of FTLD, the pathological state of FTLD in prefrontal cortical neurons is improved.

[0039] Based on this insight, the present invention demonstrates that a frontotemporal degenerative disease (FTLD) treatment agent comprising a compound represented by formula (1-1), a compound represented by formula (2-1), or a compound represented by formula (3-1), a pharmaceutically acceptable salt thereof, or a solvate thereof can treat FTLD.

[0040] More specifically, in order to solve the aforementioned problems, this application provides the following approach:

[0041] [1] A therapeutic agent for frontotemporal degenerative disease (FTLD) comprising:

[0042] a compound represented by the following formula (1-1):

[0043]

[0044] [In formula (1-1), R1 independently represents an alkyl group or a 4-hydroxyphenylethyl group having 1 to 6 carbon atoms, and n represents an integer from 1 to 3.

[0045] The compound represented by the following formula (2-1):

[0046]

[0047] [In formula (2-1), R21 is selected from the group consisting of hydrogen, fatty acid acyl groups having 2 to 18 carbon atoms, and aromatic carboxylic acid acyl groups having 7 to 9 carbon atoms];

[0048] The compound represented by the following formula (3):

[0049]

[0050] [In formula (3-1), R31 is selected from the group consisting of hydrogen, halogen, alkyl groups having 1 to 6 carbon atoms, and CF3,

[0051] R32 is selected from the group consisting of alkoxy groups having 1 to 6 carbon atoms substituted with imidazolium groups, and optionally substituted nitrogen-containing aromatic heterocycles,

[0052] R33 is selected from the group consisting of hydrogen, alkyl groups having 1 to 6 carbon atoms, and cycloalkyl groups having 3 to 6 carbon atoms,

[0053] R34 is selected from the group consisting of carboxyl groups, cyano groups, and 1H-tetrazole groups]; Specification 3 / 18 pages 8 CN 122028915 A

[0054] Their pharmaceutically acceptable salts or their solvates.

[0055] [2] The FTLD treatment agent according to claim 1, wherein FTLD is one or more symptoms selected from behavioral variant frontotemporal dementia (bvFTD), semantic dementia (SD), and progressive non-fluent aphasia (PNFA).

[0056] [3] The FTLD treatment agent according to claim 1 or 2, wherein n is 2 in the aforementioned formula (1-1).

[0057] [4] The FTLD treatment agent according to claim 1 or 2, wherein R1 is n-propyl in the aforementioned formula (1-1).

[0058] [5] The FTLD treatment agent according to claim 3, wherein R1 is n-propyl in the aforementioned formula (1-1).

[0059] [6] The FTLD therapeutic agent according to claim 1 or 2, wherein the compound represented by formula (1-1) is the compound represented by formula (1-2) below:

[0060]

[0061] that is, 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one.

[0062] [7] The FTLD therapeutic agent according to claim 1 or 2, wherein a pharmaceutically acceptable salt of the compound represented by formula (1-1) is the hydrochloride salt of the compound represented by formula (1-2) below:

[0063]

[0064] that is, 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one hydrochloride.

[0065] [8] The FTLD therapeutic agent according to [1] or [2], wherein the pharmaceutically acceptable salt of the compound shown in formula (2-1) is the compound shown in formula (2-2):

[0066]

[0067] namely 4-amino-N-(3,4-dimethyl-5-isoxazolyl)benzenesulfonamide.Instructions for Use 4 / 18 Page 9 CN 122028915 A

[0068] [9] According to [1] or [2], the pharmaceutically acceptable salt of the compound shown in formula (3-1) is the compound shown in formula (3-2) below:

[0069]

[0070] That is, 4'-[[4-methyl-6-(1-methyl-1H-benzimidazol-2-yl)-2-propyl1H-benzimidazol-1-yl]methyl]biphenyl-2-carboxylic acid.

[0071]

[10] A pharmaceutical composition for the treatment of frontotemporal degenerative disease (FTLD) containing the following substance as an active ingredient:

[0072] The compound shown in formula (1-1) below:

[0073]

[0074] [In formula (1-1), R1 independently represents an alkyl or 4-hydroxyphenylethyl group having 1 to 6 carbon atoms, and n represents an integer from 1 to 3.

[0075] The compound represented by the following formula (2-1):

[0076]

[0077] [In formula (2-1), R21 is selected from the group consisting of hydrogen, fatty acid acyl groups having 2 to 18 carbon atoms, and aromatic carboxylic acid acyl groups having 7 to 9 carbon atoms];

[0078] The compound represented by the following formula (3-1): Specification 5 / 18 page 10 CN 122028915 A

[0079]

[0080] [In formula (3-1), R31 is selected from the group consisting of hydrogen, halogen, alkyl groups having 1 to 6 carbon atoms, and CF3,

[0081] R32 is selected from the group consisting of alkoxy groups having 1 to 6 carbon atoms substituted with imidazolium groups, and optionally substituted nitrogen-containing aromatic heterocycles,

[0082] R33 is selected from the group consisting of hydrogen, alkyl groups having 1 to 6 carbon atoms, and cycloalkyl groups having 3 to 6 carbon atoms,

[0083] R34 is selected from the group consisting of carboxyl, cyano, and 1H-tetrazole groups;

[0084] pharmaceutically acceptable salts of these or their solvates.

[0085] Effects of the Invention

[0086] The FTLD therapeutic agent of the present invention is based on the analysis of prefrontal cortical neurons induced by differentiation of iPS cells derived from FTLD patients as a pathological state model, and is screened using characteristic phenotypes of human pathological states as evaluation criteria. Therefore, it is possible to provide a therapeutic agent with high efficacy against FTLD. In addition, the present invention is able to provide an FTLD therapeutic agent that is effective against three subtypes associated with the pathological state of FTLD. Brief Description of the Drawings

[0087] Figure 1 is a diagram showing the steps of prefrontal cortical neurons induced by differentiation of disease-specific iPS cells derived from FTLD patients that can be used in the screening of compounds for the treatment of FTLD.

[0088] Figure 2 is a diagram showing the creation of prefrontal cortical neurons that undergo degeneration in FTLD using disease-specific iPS cells derived from cells of FTLD patients.

[0089] Figure 3 is a diagram showing the results of drug screening using disease-specific iPS cells derived from cells of FTLD patients.

[0090] Figure 4 is a diagram showing the steps for functional analysis of ropinirole (ROPI) using prefrontal cortical neurons created using disease-specific iPS cells derived from cells of FTLD patients.

[0091] Figure 5 is a diagram showing the results of measuring the LDH leakage rate as a parameter of the neuroprotective effect of ropinirole administration, and is a diagram showing the phenotype of neuronal death (lactate dehydrogenase assay, LDH assay) (Figure 5 left) and the neuroprotective effect when ropinirole (ROPI) is administered (Figure 5 right).

[0092] Figure 6 is a graph showing the phenotype of neuronal death induced by differentiation of iPS cells from healthy individuals and iPS cells from FTLD patients (TUJ1 immunostaining) (Figure 6(a)) and the value obtained by quantifying the number of surviving neurons (Figure 6(b)).

[0093] Figure 7 is a graph showing the effect of various concentrations of ropinirole (ROPI) on the phenotype of neuronal death (TUJ1 immunostaining).

[0094] Figure 8 is a graph showing the effect of various concentrations of ropinirole (ROPI) on the phenotype of neuronal death (number of TUJ1 immunostaining positive cells) (Figure 8(a)) and the phenotype of neuronal functional recovery (neuron length) (Figure 8(b)). Specification 6 / 18 pages 11 CN 122028915 A

[0095] Figure 9 is a graph showing the effect of various concentrations of ropinirole (ROPI) on lysosomal hypertrophy.

[0096] Figure 10 is a graph showing the results of measuring the improvement rate (%) of LDH leakage in prefrontal cortical neurons induced by differentiation of iPS cells from FTLD patients with the addition of various concentrations of ropinirole.

[0097] Figure 11 is a graph showing the results of measuring the fluorescence intensity of SiR-lysosomes in prefrontal cortical neurons induced by differentiation of iPS cells from FTLD patients with the addition of various concentrations of ropinirole to the culture medium.

[0098] Figure 12 shows the results of measuring the LDH leakage rate in prefrontal cortical neurons induced by differentiation of iPS cells from FTLD patients with the addition of various concentrations of various D2 receptor agonists to the culture medium.

[0099] Figure 13 shows the results of measuring the improvement rate (%) of LDH leakage in prefrontal cortical neurons induced by differentiation of iPS cells derived from FTLD patients when various concentrations of Torin1 were added to the culture medium. Detailed Embodiments

[0100] [Screening of FTLD Therapeutic Agents and Pharmaceutical Compositions for FTLD Treatment]

[0101] In this invention, iPS cells derived from cells collected from FTLD patients were prepared, and prefrontal cortical neurons were induced by differentiation of these iPS cells. Using prefrontal cortical neurons exhibiting the pathological state of FTLD, compounds in a compound library were screened for improvement of lysosomal function and neuroprotective effects (increased neuronal survival) as evaluation criteria. This resulted in compounds that could improve the pathological state of FTLD in prefrontal cortical neurons and had a therapeutic effect on FTLD.

[0102] The method for inducing differentiation of prefrontal cortical neurons can be as follows: after inducing differentiation of iPS cells into the brain nervous system by dual SMAD inhibition (e.g., LDN-193189 and SB431542) and Wnt inhibition (e.g., XAX939), Fgf8b is added (Non-Patent Literature 2). The inventors conducted a study and the results showed that dual SMAD inhibition alone cannot induce iPS cells to differentiate into the brain nervous system. In addition, even with Wnt antagonist treatment on the basis of dual SMAD inhibition, differentiation into the brain nervous system cannot be induced, showing that dual SMAD inhibition and Wnt inhibition are necessary for inducing differentiation into the brain nervous system.

[0103] The compound library used in the screening can be of any type, for example, a compound library that uses compounds that have undergone clinical trials for various diseases and whose safety has been confirmed as the library.

[0104] In this invention, the result of screening the compound library by this method is that ropinirole, telmisartan, and sulfisoxazole can be selected as compounds that improve lysosomal function and exhibit neuroprotective effects (increased neuronal survival). Based on this insight, the inventors of this invention completed this invention.

[0105] [FTLD therapeutic agent, FTLD therapeutic pharmaceutical composition]

[0106] In this invention, the compound represented by the following formula (1-1) is used:

[0107]

[0108] [In formula (1-1), R1 independently represents an alkyl or 4-hydroxyphenylethyl group having 1 to 6 carbon atoms, and n represents an integer of 1 to 3. (See page 7 / 18 of specification, CN 122028915 A)

[0109] The compound represented by the following formula (2-1):

[0110]

[0111] [In formula (2-1), R21 is selected from the group consisting of hydrogen, fatty acid acyl groups having 2 to 18 carbon atoms, and aromatic carboxylic acid acyl groups having 7 to 9 carbon atoms];

[0112] The compound represented by the following formula (3-1):

[0113]

[0114] [In formula (3-1), R31 is selected from the group consisting of hydrogen, halogen, alkyl groups having 1 to 6 carbon atoms, and CF3,

[0115] R32 is selected from the group consisting of alkoxy groups having 1 to 6 carbon atoms substituted with imidazolium groups, and optionally substituted nitrogen-containing aromatic heterocycles,

[0116] R33 is selected from the group consisting of hydrogen, alkyl groups having 1 to 6 carbon atoms, and cycloalkyl groups having 3 to 6 carbon atoms,

[0117] R34 is selected from the group consisting of carboxyl groups, cyano groups, and 1H-tetrazole groups];

[0118] The present invention provides, regarding the pharmaceutical use of their pharmaceutically acceptable salts or solvates, an FTLD therapeutic agent comprising the above-described compounds, their pharmaceutically acceptable salts or solvates; or an FTLD therapeutic pharmaceutical composition comprising the above-described compounds, their pharmaceutically acceptable salts or solvates.

[0119] As an FTLD disease that can be treated with the FTLD therapeutic agent or the FTLD therapeutic pharmaceutical composition of the present invention, any of behavioral variant frontotemporal dementia (bvFTD), semantic dementia (SD), and progressive non-fluent aphasia (PNFA) may also be included. In addition, based on the pathological characteristics of patients with FTLD, any of the following FTLD patients can be included as treatment subjects: patients with confirmed TAU accumulation FTLD-tau, patients with confirmed TDP-43 accumulation FTLD-TDP43, and patients with confirmed FUS accumulation FTLD-FUS. Furthermore, based on the genetic characteristics of patients with FTLD, any patient with FTLD, such as those with GRN gene mutations, MAPT gene mutations, C9ORF72 gene mutations, or TARDBP gene mutations, can be a treatment target.

[0120] In the compounds of formula (1-1) above, which are the active ingredients of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention, n in formula (1-1) can be 1, 2, or 3. In the case where the active ingredient of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention is ropinirole as described below, n is 2 in formula (1-1). Therefore, the active ingredient of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention can also be a compound in formula (1-1) where n is 2.

[0121] Furthermore, in the compound described in the above formula (8 / 18 pages, CN 122028915 A (1-1) of the specification, which is the effective component of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention, R1 in the above formula (1-1) can be a straight-chain, branched, or cyclic alkyl group having 1 to 6 carbon atoms. More specifically, examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, cyclopentyl, n-hexyl, cyclohexyl, etc. In the case where the effective component of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention is ropinirole as described below, R1 in formula (1-1) is n-propyl. Therefore, the effective component of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention can also be a compound in the above formula (1-1) where R1 is n-propyl.

[0122] The compound shown in formula (1-1) above, which is an effective component of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention, may also be 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one. That is, the compound shown in formula (1-1) above may also be ropinirole. The chemical formula of ropinirole is shown in the following formula (1-2).

[0123]

[0124] Ropinirole originally had dopamine D2 receptor agonist activity of dopamine neurons and was therefore developed as a treatment for Parkinson's disease. In the present invention, it is not currently known whether ropinirole exerts its effect on FTLD based on the same intracellular mechanism of action or based on a different intracellular mechanism of action. However, clinical trials as a drug have been completed, and its safety when administered to organisms has been fully confirmed. Thus, since ropinirole is an existing drug, it is possible to rapidly develop FTLD therapeutic agents or FTLD therapeutic pharmaceutical compositions.

[0125] The compound shown in formula (2-1) above, which is an effective component of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention, may also be 4-amino-N-(3,4-dimethyl-5-isoxazolyl)benzenesulfonamide. That is, the compound shown in formula (2-1) above may also be sulfamethoxazole. The chemical formula of sulfamethoxazole is shown in the following formula (2-2).

[0126]

[0127] Sulfamethoxazole was originally developed as a sulfonamide antibacterial drug, but in recent years, its application in the treatment of diseases such as Alzheimer's disease and Parkinson's disease based on modified cellular stress responses has been studied. In the present invention, it is currently unclear whether sulfamethoxazole exerts its effect on FTLD based on the same intracellular mechanism of action or based on a different intracellular mechanism of action. However, clinical trials as a drug have been completed, and its safety when administered to organisms has been fully confirmed.Thus, since sulfamethoxazole is an existing drug, FTLD therapeutic agents or FTLD therapeutic drug compositions can be rapidly developed. Specification 9 / 18 pages 14 CN 122028915 A

[0128] For the compound shown in formula (3-1) above as an active ingredient of the FTLD therapeutic agent or FTLD therapeutic drug composition of the present invention,

[0129] the compound shown in formula (3-1) above as an active ingredient of the FTLD therapeutic agent or FTLD therapeutic drug composition of the present invention may also be 4'-[[4-methyl-6-(1-methyl-1H-benzimidazol-2-yl)-2-propyl1H-benzimidazol-1-yl]methyl]biphenyl-2-carboxylic acid. That is, the compound shown in formula (3-1) above may also be telmisartan. The chemical formula of telmisartan is shown in the following formula (3-2).

[0130]

[0131] Telmisartan originally possessed bile-excreting sustained AT1 receptor blocker activity and was therefore developed as a treatment for hypertension. In this invention, it is currently unclear whether telmisartan exerts its effect on FTLD based on the same intracellular mechanism of action or on a different intracellular mechanism. However, clinical trials as a drug have been completed, and its safety when administered to organisms has been well established. Thus, since telmisartan is an existing drug, FTLD therapeutic agents or FTLD therapeutic drug compositions can be rapidly developed.

[0132] The active ingredient of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention may be a salt of any of the compounds shown in formula (1-1), formula (2-1), or formula (3-1), or a solvate of any of the compounds shown in formula (1-1), formula (2-1), or formula (3-1), or a solvate of a salt of any of the compounds shown in formula (1-1), formula (2-1), or formula (3-1).

[0133] In the effective components of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention, when using a salt of any of the compounds shown in formula (1-1), formula (2-1), or formula (3-1), there are no particular limitations as long as the salt is pharmaceutically acceptable. Examples include inorganic acid salts such as hydrochloride, sulfate, hydrobromide, nitrate, and phosphate; organic acid salts such as acetate, methanesulfonate, succinate, maleate, fumarate, citrate, and tartrate; alkali metal salts such as sodium and potassium salts; alkaline earth metal salts such as magnesium and calcium salts; metal salts such as aluminum and zinc salts; ammonium salts such as ammonium salts and tetramethylammonium salts; organic amine addition salts such as morpholine and piperidine; and amino acid addition salts such as glycine, phenylalanine, lysine, aspartic acid, and glutamic acid.

[0134] Furthermore, in the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention, when using a solvate of any of the compounds shown in formula (1-1), formula (2-1), or formula (3-1) above, or a salt thereof, there are no particular limitations as long as the solvate is pharmaceutically acceptable; for example, hydrates, organic solvates, etc., can be listed.

[0135] The active ingredient of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention may also be 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one hydrochloride, i.e., ropinirole hydrochloride.

[0136] The active ingredient of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention may also be 4-amino-N-(3,4-dimethyl-5-isoxazolyl)benzenesulfonamide, i.e., sulfamethoxazole.

[0137] The active ingredient of the FTLD therapeutic agent or FTLD therapeutic pharmaceutical composition of the present invention may also be 4'-[[4-methyl-6-(1-methyl-1H-benzimidazol-2-yl)-2-propyl1H-benzimidazol-1-yl]methyl]biphenyl-2-carboxylic acid, i.e., telmisartan.

[0138] The FTLD therapeutic pharmaceutical composition of the present invention can be formulated in the form of a pharmaceutical composition, for example, it can be administered orally in the form of tablets, capsules, elixirs, microcapsules, etc., or it can be administered non-orally in the form of injections, suppositories, topical skin preparations, etc. More specifically, ointments, patches, etc. can be listed as topical skin preparations.

[0139] In the FTLD therapeutic pharmaceutical composition of the present invention, a pharmaceutically acceptable carrier may be used without particular limitation, such as a carrier commonly used in pharmaceutical compositions.More specifically, examples include binders such as hydroxypropyl methylcellulose, dextrin, polyethylene glycol 400, gelatin, corn starch, tragacanth gum, and gum arabic; excipients such as lactose hydrate, D-mannitol, starch, crystalline cellulose, and alginic acid; solvents for injection such as water, ethanol, and glycerin; and adhesives such as rubber-based adhesives and silicone-based adhesives.

[0140] The FTLD therapeutic pharmaceutical composition of the present invention may contain additives. Examples of additives include lubricants such as calcium stearate and magnesium stearate; sweeteners such as sucrose, lactose, saccharin, and maltitol; flavorings such as peppermint and red beech oil; stabilizers such as sodium carboxymethyl cellulose, hardened oil, light anhydrous silica, povidone, glyceryl fatty acid esters, benzyl alcohol, and phenol; buffers such as phosphates and sodium acetate; cosolvents such as benzyl benzoate and benzyl alcohol; and colorants such as iron oxide yellow, ferric oxide, iron oxide black, and titanium oxide.

[0141] The FTLD therapeutic pharmaceutical composition of the present invention can be formulated by appropriately combining the above-mentioned active ingredient with the above-mentioned pharmaceutically acceptable carrier and additives, and mixing them in a unit dosage form required by generally recognized pharmaceutical practice. The active ingredient of the FTLD therapeutic pharmaceutical composition of the present invention can be used alone or in combination with two or more.

[0142] Generally, the suitable daily dosage of the FTLD therapeutic pharmaceutical composition of the present invention is the amount of active ingredient contained in the minimum effective dosage for producing a therapeutic effect. The above-mentioned effective minimum dosage depends on various factors including: the activity of the active ingredient contained in the FTLD therapeutic pharmaceutical composition, the specified fat-soluble / water-soluble functional group modification, the route of administration, the time of administration, the excretion rate of the specific active ingredient used, the treatment period, other drugs, compounds and / or substances used in combination, age, sex, weight, disease, health status and the patient's pre-existing condition, and other factors known in the medical field. Generally, the dosage of the FTLD therapeutic pharmaceutical composition of the present invention for a patient is the amount of active ingredient containing about 0.0001 to about 100 mg / kg body weight per day. The FTLD therapeutic pharmaceutical composition of the present invention can be administered once a day or in 2 to 4 divided doses.

[0143] In particular, when the active ingredient is a compound represented by formula (1-2), for the dosage of the FTLD therapeutic pharmaceutical composition of the present invention, for example, if it is a ropinirole sustained-release formulation, it is possible to consider oral administration of 2 mg of the active ingredient once a day, increasing the dose weekly, within the range of no more than 16 to 24 mg of the active ingredient per day.

[0144] In particular, when the active ingredient is a compound represented by formula (2-2), for the dosage of the FTLD therapeutic pharmaceutical composition of the present invention, for example, if it is sulfasoxazole, it is possible to consider oral administration of 4,000 mg of the active ingredient 4 to 6 times a day, gradually increasing the dose, within the range of not exceeding 8,000 mg of the active ingredient per day.

[0145] In particular, when the active ingredient is a compound represented by formula (3-2), for the dosage of the FTLD therapeutic pharmaceutical composition of the present invention, for example, if it is telmisartan extended-release, it is possible to consider oral administration of 20 mg of the active ingredient once a day, gradually increasing the dose, within the range of not exceeding 80 mg of the active ingredient per day.

[0146] [Other Embodiments] Specification 11 / 18 pages 16 CN 122028915 A

[0147] In another aspect, the present invention provides a treatment method for FTLD, comprising the steps of administering any one of the compounds shown in formula (1-1), formula (2-1), or formula (3-1), a pharmaceutically acceptable salt thereof, or a solvate thereof to a patient requiring treatment for FTLD. In this aspect of the present invention, the same substances as described above can be used as the active ingredient, such as the compound shown in formula (1-1), formula (2-1), or formula (3-1), a pharmaceutically acceptable salt thereof, or a solvate thereof. Furthermore, in this aspect of the present invention, the dosage of the active ingredient can be determined based on suitable dosage studies for each compound, for example, as described above when ropinirole is used as the active ingredient.

[0148] In this invention, any one of the compounds shown in formula (1-1), formula (2-1), or formula (3-1) above, a pharmaceutically acceptable salt thereof, or a solvate thereof is provided for treating FTLD. In this aspect of the invention, the same substances as described above can be used as any one of the compounds shown in formula (1-1), formula (2-1), or formula (3-1) above, a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0149] Hereinafter, examples are given to specifically illustrate the invention. The examples shown below are not intended to limit the invention in any way.

[0150] Examples

[0151] Example 1: Screening of compounds for the treatment of FTLD

[0152] In this example, disease-specific iPS cells derived from FTLD patients were used to differentiate / induce prefrontal cortical neurons that could be used in the screening of compounds for the treatment of FTLD, and to screen for compounds that could improve the pathological state of FTLD in these prefrontal cortical neurons.

[0153] (1-1) Differentiation / Induction of Prefrontal Cortical Neurons

[0154] It is known that familial FTLD is caused by GRN gene mutations, MAPT gene mutations, and C9ORF72 gene mutations. Therefore, based on the culture method shown in Figure 1, iPS cells derived from healthy individuals and iPS cells derived from FTLD patients with GRN gene mutations (bvFTD patients with GRNS116X mutations), who are representatives of familial FTLD, were differentiated into prefrontal cortical neurons. The mutation of the GRN gene is S116X.

[0155] Specifically, firstly, in a culture medium containing LDN-193189 (CAS No.: 1062368-24-4) at a final concentration of 50 nM, SB431542 (CAS No.: 301836-41-9) at a final concentration of 2 μM, and XAV939 (CAS No.: 284028-89-3) at a final concentration of 1 μM, each cell line (Ngn2-iPSC in Figure 1) containing a plasmid ligated to a promoter that can be inducibly expressed in the presence of DOX via the PiggyBac method was cultured for 6 days to induce cortical neural stem cells. The culture medium was changed every 2-3 days.

[0156] Next, the obtained cortical neural stem cells were isolated one by one and cultured in a culture medium containing Fgf8b for 3 days.

[0157] Subsequently, while inducing the expression of the transcription factor NGN2 protein, which is expressed in the early stage of neuronal differentiation, with doxycycline (CAS No.: 24390-14-5) at a final concentration of 0.25 ng / mL, the cells were cultured for 5 days in a medium containing DAPT (CAS No.: 208255-80-5) at a final concentration of 3 μM and palbociclib (CAS No.: 571190-30-2) at a final concentration of 2 μM. The medium was changed every 2 to 3 days.

[0158] Differentiation-induced prefrontal cortical neurons were immunostained using anti-Beta III Tubulin Antibody (Chemicon) and anti-Pea3 antibody (Abcam) to investigate the expression of βIII-tubulin and polyomavirus enhancer activator 3 (Pea3). The results are shown in Figure 2. As a result, for the obtained neurons, PEA3, a prefrontal marker, was confirmed to be expressed using anti-Pea3 antibody (Abcam) on D6 (according to the culture days in Figure 1), and TUJ1, a pan-neuronal marker, was confirmed to be expressed using anti-Tubulin βIII antibody (Chemicon) on D13 (according to the culture days in Figure 1). This confirmed that the obtained cells were prefrontal cortical neurons.

[0159] (1-2) Results of drug screening using prefrontal cortical neurons

[0160] Using prefrontal cortical neurons derived from FTLD disease-specific iPS cells derived from the differentiation-induced FTLD disease strain (bvFTD patients with GRNS116X mutation) in Example 1 (1-1), drugs that restore the phenotype of FTLD (close to wild type) were screened from an existing drug library using lactate dehydrogenase (LDH) leakage rate and TUJ1 positive cell count as markers of neuronal death, and the fluorescence intensity of LAMP1 immunostaining as a marker of lysosomal abnormalities. The LDH leakage rate was determined using a commercially available kit (model "G7891", Promega). The number of TUJ1 positive cells was determined by immunostaining with anti-tubulin β-III antibody (Chemicon), and LAMP1 immunostaining as a marker of lysosomal abnormalities was performed using anti-LAMP1 antibody [H4A3] (Abcam).

[0161] After adding compounds from the existing drug library to the culture medium from days 13 to 17 from the start of differentiation induction, screening was performed, and three drugs, namely ropinirole, telmisartan, and sulfisoxazole, were found to be promising therapeutic agents for FTLD. Figure 3 shows the improvement rate (%) of FTLD phenotype when these drugs were added to the culture medium.

[0162] The improvement rate (%) of the FTLD phenotype was calculated for each parameter using the following formula (1):

[0163] Improvement rate (%) = (A-B) / (A-C) × 100…(1)

[0164] [In formula (1),

[0165] A represents the measured value of prefrontal cortical neurons induced by iPS cell differentiation from FTLD patients in the absence of drug,

[0166] B represents the measured value of prefrontal cortical neurons induced by iPS cell differentiation from FTLD patients in the presence of drug,

[0167] C represents the measured value of prefrontal cortical neurons induced by iPS cell differentiation from healthy individuals in the absence of drug. ]

[0168] Figure 3 shows the results of adding the three drugs (ropinirole, telmisartan, and sulfamethoxazole) obtained above to a final concentration of 100 nM, 1 μM, and 10 μM to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells derived from FTLD patients. The results show that all three drugs have neuroprotective effects and lysosomal function improvement effects in the range of at least 100 nM to 10 μM. For ropinirole of these three drugs, further studies were conducted in the following examples.

[0169] Example 2: Effects of ropinirole

[0170] In this example, functional analysis was performed on prefrontal cortical neurons made from disease-specific iPS cells derived from cells of FTLD patients using ropinirole obtained in Examples (1-2).

[0171] (2-1) Steps for treating cells with ropinirole

[0172] The timing of adding ropinirole to the culture was changed to days 8 to 13 from the start of differentiation induction, as shown in Figure 4. Otherwise, the efficacy of ropinirole was evaluated in the same manner as in Examples 1-2.

[0173] Specifically, ropinirole was added to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells from patients with GRN gene mutations in FTLD (GRN-FTLD), on days 8 and 10, and the results were tested on day 13. Ropinirole was added to the culture medium at final concentrations of 10 nM, 30 nM, 100 nM, and 300 nM.

[0174] (2-2) Neuroprotective effect of ropinirole, 13 / 18 pages, 18 CN 122028915 A

[0175] As a parameter of neuroprotective effect, the LDH leakage rate based on ropinirole administration was measured. The LDH leakage rate was determined using the same commercially available kit (model "G7891", Promega) as in Examples 1 (1-2). The results are shown in Figure 5.

[0176] Figure 5 on the left shows the results of measuring the LDH leakage rate of prefrontal cortical neurons induced by differentiation of iPS cells from healthy individuals and iPS cells from GRNS116XFTLD patients. On the left side of Figure 5, the vertical axis represents the LDH leakage rate (relative value) of prefrontal cortical neurons induced by differentiation of iPS cells from FTLD patients, calculated based on the average LDH leakage rate in prefrontal cortical neurons induced by differentiation of iPS cells from healthy individuals. “ ”, “##”, and “&&” indicate that there is a significant difference between GRNS116XFTLD patient-derived cerebral cortical neurons and healthy individual-derived cerebral cortical neurons (RC802, ND025, ND554) at a significance level of less than 1%.

[0177] In contrast, the right side of Figure 5 shows the results of LDH leakage rate determination when various concentrations (30 nM, 100 nM, 300 nM, 1 μM) of ropinirole were added to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells derived from GRNS116XFTLD patients. On the right side of Figure 5, the vertical axis represents the LDH leakage rate (relative value) calculated based on the LDH leakage without ropinirole (0 nM), and the horizontal axis represents the results of adding ropinirole to the culture medium at any of the amounts of 30 nM, 100 nM, 300 nM, and 1 μM. On the right side of Figure 5, “ ” indicates a significant difference at a significance level of less than 1%.

[0178] As a result, even from day 8 to 13 from the start of differentiation induction, a neuroprotective effect (reduction in LDH leakage) was confirmed by the addition of ropinirole. In addition, ropinirole's EC50 = 38.1 nM (calculated using ImageJ and Excel) shows that it can exert a neuroprotective effect at very low doses.

[0179] (2-3) Effect of ropinirole on neuronal death

[0180] First, the phenotype of neuronal death was investigated between prefrontal cortical neurons induced by differentiation of iPS cells from healthy individuals and those induced by differentiation of iPS cells from FTLD patients (TUJ1 immunostaining), using TUJ1 expression as a parameter. TUJ1 immunostaining was performed using an anti-microtubule β-III antibody (Chemicon) in the same manner as in Example 1 (1-1). The results are shown in Figure 6.

[0181] Specifically, Figure 6 shows the results of TUJ1 immunostaining of prefrontal cortical neurons induced by differentiation of iPS cells from healthy individuals and iPS cells from FTLD patients (Figure 6(a)), and a graph showing the values ​​obtained by quantifying the number of surviving neurons based on this (Figure 6(b)).In Figure 6(b), the vertical axis represents the number of TUJ1-positive cells in prefrontal cortical neurons induced by differentiation of iPS cells from healthy individuals (relative value), calculated based on the average number of TUJ1-positive cells in prefrontal cortical neurons induced by differentiation of iPS cells from healthy individuals. " ", "##, and "&" indicate significant differences between GRNS116XFTLD patient-derived prefrontal cortical neurons and healthy individual-derived prefrontal cortical neurons (RC802, ND025, ND554) at significance levels below 1%, below 1%, and below 5%, respectively.

[0182] As a result, as a characteristic of prefrontal cortical neurons induced by differentiation of iPS cells, neuronal death (a decrease in the number of TUJ1-positive cells) was observed in prefrontal cortical neurons induced by differentiation of iPS cells from GRNS116XFTLD patient-derived prefrontal cortical neurons from days 8 to 13 from the start of differentiation induction.

[0183] Next, ropinirole was added to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells from patients with GRN gene mutations (GRNS116XFTLD) from day 8 to 13 from the start of differentiation induction. The number of TUJ1-positive cells, a parameter of neuronal death, and the length of neurites, a parameter of neuronal function, were measured. Ropinirole was added to the culture medium at final concentrations of 30 nM, 100 nM, 300 nM, and 1 μM. The results are shown in Figures 7 and 8.

[0184] Figure 7 is a staining image showing the TUJ1 immunostaining results when ropinirole was added to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells from patients with GRNS116XFTLD. “ROPI” indicates the results of adding ropinirole (page 14 / 18 of the specification, CN 122028915 A) to the culture medium at various concentrations. The results show that ropinirole has a neuroprotective effect.

[0185] In addition, the characteristics of prefrontal cortical neurons were investigated by adding various concentrations of ropinirole to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells derived from GRNS116XFTLD patients, using the number of TUJ1 positive cells and neurite length as parameters. The results are shown in Figure 8.

[0186] Here, Figure 8(a) is a graph showing the results of measuring the number of TUJ1 positive cells when various concentrations of ropinirole were added to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells derived from GRNS116XFTLD patients. In Figure 8(a), the vertical axis represents the number of TUJ1 positive cells (relative value) calculated based on the number of TUJ1 positive cells without the addition of ropinirole (0 nM).“ROPI” indicates the results of adding ropinirole to the culture medium at various concentrations.

[0187] Figure 8(b) is a graph showing the results of neurite length measurements when various concentrations of ropinirole were added to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells derived from GRNS116XFTLD patients. In Figure 8(b), the vertical axis represents the neurite length (relative value) calculated based on the neurite length without ropinirole (0 nM). “ROPI” indicates the results of adding ropinirole to the culture medium.

[0188] Based on these results, from day 8 to 13 from the start of differentiation induction, the addition of ropinirole confirmed neuroprotective effects (increase in the number of TUJ1 positive cells) and the functional recovery of nerve cells (increase in neurite length). That is, the neuroprotective effect of ropinirole was also confirmed by methods other than the LDH assay.

[0189] (2-4) Effect of ROPI on lysosomal hypertrophy

[0190] It is known that nerve cells in FTLD disease are characterized by lysosomal hypertrophy, so the effects of ropinironiro on lysosomal hypertrophy were investigated. The timing of ropinironiro addition was changed to days 8-13 from the start of differentiation induction. Otherwise, immunostaining against LAMP1, a marker of lysosomal abnormality, was performed in the same manner as in (1-2) of Example 1, thereby evaluating the efficacy of ropinironiro.

[0191] Specifically, various concentrations of ropinironiro were added to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells from FTLD patients with GRN gene mutation (S116X) (GRNS116XFTLD), and the fluorescence intensity of LAMP1 immunostaining, a membrane protein of the lysosomal body membrane, was measured. The determination of the fluorescence intensity of LAMP1 immunostaining was performed using anti-LAMP1 antibody [H4A3] (Abcam) in the same manner as in Examples 1-2. Ropinirole was added to the culture medium at final concentrations of 10 nM, 30 nM, 100 nM, and 300 nM. The results are shown in Figure 9.

[0192] Figure 9 is a graph showing the determination of the fluorescence intensity of LAMP1 immunostaining when various concentrations of ropinirole were added to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells from FTLD patients. In Figure 9, the vertical axis represents the LAMP1 fluorescence intensity (relative value) calculated based on the LAMP1 fluorescence intensity without the addition of ropinirole (0 nM). The horizontal axis represents the concentration of ropinirole added. “ROPI” indicates the result of adding ropinirole to the culture medium. In Figure 9, “ ” indicates a significant difference at a significance level of less than 1%.

[0193] As a result, even on days 8–13 from the start of differentiation induction, the addition of ropinirole confirmed an improvement in lysosomal function (a decrease in LAMP1 fluorescence intensity), with an EC50 of 16.7 nM for ropinirole.

[0194] Example 3: Study using various FTLD strains

[0195] In this example, the effect of ropinirole on neuronal cell death was investigated using disease-specific iPS cells from patients with FTLD of various causes.

[0196] (3-1) Inhibitory effect of ropinirole on neuronal death

[0197] Using the same method as in Example 1 (1-2), iPS cells from multiple familial (gene mutation) FTLD patients (GRNM1LFTLD, GRNS116XFTLD, GRNR493XFTLD, MAPTR406WFTLD, 1 case each) and iPS cells from sporadic FTLD patients (SD 2 cases (sFTD1, sFTD2), bvFTD 1 case (sFTD3), and 1 case with unknown clinical symptoms (sFTD4)) were differentiated into prefrontal cortical neurons. In the culture medium of various prefrontal cortical neurons induced by differentiation of iPS cells derived from FTLD patients, various concentrations of ropinirole were added from day 8 to 13 from the start of differentiation induction. The LDH leakage rate, a parameter of neuronal death, was measured using a commercially available kit (model "G7891", Promega). Ropinirole was added to the culture medium at final concentrations of 10 nM, 30 nM, 100 nM, and 300 nM. LDH leakage was measured in n=3 to 5 measurements under each experimental condition. Based on these results, the LDH leakage improvement rate (relative value) was calculated from the LDH leakage value before ropinirole administration (0 nM) to the LDH leakage value after ropinirole administration (300 nM), using the LDH leakage value in healthy human control samples as a baseline (100%). The calculation of the LDH leakage improvement rate was performed in the same manner as in Examples 1 (1-2). The results are shown in Figure 10.

[0198] Figure 10 is a graph showing the results of the determination of the improvement rate (%) of LDH leakage when various concentrations of ropinirole were added to the culture medium of prefrontal cortical neurons induced by differentiation of iPS cells from FTLD patients. In Figure 10, “ ” and “ ” indicate that there were significant differences at a significance level of less than 5% or less than 1%, respectively, by one-way ANOVA based on Tukey post-hoc test.

[0199] As a result, neuroprotective effects (reduction of LDH leakage) were also confirmed by the addition of ropinirole in prefrontal cortical neurons derived from iPS cells in multiple FTLD diseases.

[0200] This result supports the view that ropinirole is effective not only against the S116X mutation of the GRN gene, but also against familial (gene mutation) FTLD and sporadic FTLD.

[0201] (3-2) Effect of ROPI on CatD activity

[0202] Since lysosomal function is abnormal in nerve cells of FTLD, the activity of cathepsin D, one of the main protein-degrading enzymes present in lysosomes, was investigated by adding ropinirole, thereby analyzing the mechanism of action of ropinirole. In this experiment, the timing of ropinirole addition was changed to day 8 to 13 from the start of differentiation induction, and the ropinirole treatment was performed in the same manner as in Example 1 (1-2).

[0203] Specifically, for prefrontal cortical neurons induced by differentiation of iPS cells from FTLD patients with GRN gene mutation (S116X) (GRNS116XFTLD), various concentrations of ropinirole were added to the culture medium, and the fluorescence intensity of the fluorescent dye SiR-lysosome kit (Cytoskeleton Inc.) was measured after CatD digestion. The fluorescence intensity of SiR-lysosomes was measured in the same manner as in (1-2) of Example 1. Ropinirole was added to the culture medium at final concentrations of 10 nM, 30 nM, 100 nM, and 300 nM, and the n=4 values ​​were measured for each concentration. The results are shown in Figure 11.

[0204] Figure 11 is a graph showing the results of the fluorescence intensity measurement of SiR-lysosomes when various concentrations of ropinirole were added to the culture medium for prefrontal cortical neurons induced by differentiation of iPS cells from FTLD patients. In Figure 11, the vertical axis represents the relative fluorescence intensity of SiR-lysosomes calculated based on the fluorescence intensity of SiR-lysosomes without ropinirole (0 nM). The horizontal axis represents the concentration of added ropinirole. “ROPI” indicates the result of adding ropinirole to the culture medium. In Figure 11, “ ” indicates that a significant difference exists at a significance level of less than 5% in a one-way ANOVA based on Tukey’s post-hoc test.

[0205] As a result, even from day 8 to 13 from the start of differentiation induction, the addition of ropinirole confirmed an improvement in CatD activity (an increase in SiR-lysosome fluorescence intensity).

[0206] (3-3) Mechanism of neuroprotective effect of ropinirole analyzed by DRD2 dependence / independence analysis

[0207] It is known that ropinirole originally has dopamine D2 receptor (D2R) agonist activity of dopamine neurons. Therefore, the neuroprotective effects of bromocriptine, rotigotine, and sumanirole, which are also D2 receptor agonists, were compared with those of ropinirole.

[0208] As a parameter of neuroprotective effect, the LDH leakage rate after administration of ropinirole or various D2 receptor agonists (bromocriptine, rotigotine, sumanirole) was measured. The LDH leakage rate was measured using a commercially available kit (model "G7891", Promega) in the same manner as in Example 1 (1-2). The results are shown in Figure 12.

[0209] Figure 12 is a graph showing the results of LDH leakage rate determination for prefrontal cortical neurons induced by differentiation of iPS cells derived from GRNS116XFTLD patients, with the addition of various concentrations (10 nM, 30 nM, 100 nM, 300 nM) of bromocriptine, rotigotine, or sumanilol to the culture medium. On the right side of Figure 5, the vertical axis represents the LDH leakage rate (relative value) calculated based on the LDH leakage without the addition of each substance (0 nM), and the horizontal axis represents the results obtained by adding any of bromocriptine, rotigotine, or sumanilol to the culture medium at any amount of 10 nM, 30 nM, 100 nM, or 300 nM.

[0210] As a result, from day 8 to day 13 from the start of differentiation induction, the neuroprotective effect (reduction in LDH leakage) observed with the addition of ropinirol was not confirmed in the presence of various D2 receptor agonists.

[0211] (3-4) Analysis of the neuroprotective mechanism of ropinirole by using mTOR inhibitors

[0212] The inventors predicted that ropinirole would exert its effect of inhibiting neuronal cell death by inhibiting mTOR itself or its signaling pathway. To confirm this effect, it is preferable that the neuronal cell death inhibition effect of ropinirole disappears when an mTOR activator is added, but there is a problem that there is a lack of suitable mTOR activators. Therefore, in order to help clarify such a mechanism of action of ropinirole, it was decided to indirectly infer that ropinirole inhibits mTOR based on whether the mTOR inhibitor has the same neuronal cell death inhibition effect as ropinirole.

[0213] In this technical field, rapamycin is known as an mTOR inhibitor, but it is known to only inhibit a part of the function of mTOR (i.e., S6K, a protein downstream of mTOR, can be completely inhibited, but another downstream protein, 4EBP, cannot be completely inhibited).This is believed to be due to the mechanism of mTOR inhibition by rapamycin.

[0214] On the other hand, it is known that second-generation mTOR inhibitors competitively bind to the ATP binding site required for the kinase activity of mTOR, and thus can almost completely inhibit the kinase activity of mTOR. Therefore, in this embodiment, the second-generation mTOR inhibitor TORIN1 was used in the experiment.

[0215] As a parameter of neuroprotective effect, the LDH leakage rate based on nerve cell death after administration of Torin1 was measured. Torin1 has the following chemical formula.

[0216]

[0217] For the determination of LDH leakage, n=3 measurements were performed for each experimental condition. Based on the measurement results, the LDH leakage improvement rate (relative value) from the LDH leakage value before administration of Torin1 (0 nM) to the LDH leakage value after administration of Torin1 (300 nM) was calculated with the LDH leakage value in the healthy human control sample as a baseline (100%). The determination of LDH leakage rate was performed using a commercially available kit (model "G7891", Promega) in the same manner as in Examples 1 (1-2). The results are shown in Figure 13.

[0218] Figure 13 is a graph showing the results of the determination of the LDH leakage improvement rate (%) for prefrontal cortical neurons induced by differentiation of iPS cells derived from GRNS116XFTLD patients when various concentrations (3nM, 10nM, 30nM, 100nM) of Torin1 (Selleck) were added to the culture medium. In Figure 13, " " indicates that there was a significant difference at a significance level of less than 5% by one-way ANOVA based on Tukey post-hoc test.

[0219] As a result, from day 8 to 13 from the start of differentiation induction, the addition of Torin1 confirmed a neuroprotective effect (reduction of LDH leakage) comparable to that of ropinirole. Therefore, the mechanism of action of ropinirole is presumed to be mediated through mTOR inhibition.

[0220] Industrial Applicability

[0221] According to the present invention, FTLD therapeutic agents and FTLD therapeutic compositions can be provided. The FTLD therapeutic agents or FTLD therapeutic compositions of the present invention can treat not only familial FTLD but also sporadic FTLD. Furthermore, by analyzing the pharmacodynamic mechanism of the FTLD therapeutic agents of the present invention on prefrontal cortical neurons differentiated from iPS cells derived from FTLD patients, the pathological mechanism of FTLD can be elucidated.Instruction manual 18 / 18 page 23 CN 122028915 A Figure 1 Figure 2 Figure 3 Instruction manual drawing 1 / 7 page 24 CN 122028915 A Figure 4 Figure 5 Instruction manual drawing 2 / 7 page 25 CN 122028915 A Figure 6 Instruction manual drawing 3 / 7 page 26 CN 122028915 A Figure 7 Figure 8 Instruction manual drawing 4 / 7 page 27 CN 122028915 A Figure 9 Figure 10 Instruction manual drawing 5 / 7 page 28 CN 122028915 A Figure 11 Figure 12 Instruction manual drawing 6 / 7 page 29 CN 122028915 A Figure 13 Instruction manual drawing 7 / 7 page 30 CN 122028915 A.

Claims

1. A therapeutic agent for frontotemporal degenerative disease (FTLD), comprising: The following compound is represented by formula (1-1): In equation (1-1), R 1 Each of the following can be independently represented as an alkyl group or a 4-hydroxyphenylethyl group having 1 to 6 carbon atoms, where n represents an integer from 1 to 3; The following compound is represented by formula (2-1): In equation (2-1), R 21 The group consisting of hydrogen, fatty acid acyl groups with 2 to 18 carbon atoms, and aromatic carboxylic acid acyl groups with 7 to 9 carbon atoms; The following compound is represented by formula (3): In equation (3-1), R 31 Choose from the group consisting of hydrogen, halogens, alkyl groups with 1 to 6 carbon atoms, and CF3. R 32 The group consisting of alkoxy groups with 1 to 6 carbon atoms substituted by imidazole groups and optionally substituted nitrogen-containing aromatic heterocycles is selected. R 33 The group consisting of hydrogen, alkyl groups having 1 to 6 carbon atoms, and cycloalkyl groups having 3 to 6 carbon atoms is selected. R 34 Select from the group consisting of carboxyl, cyano, and 1H-tetrazole groups; Their pharmaceutically acceptable salts or their solvates.

2. The FTLD therapeutic agent according to claim 1, wherein, FTLD is one or more symptoms selected from behavioral variant frontotemporal dementia (bvFTD), semantic dementia (SD), and progressive non-fluent aphasia (PNFA).

3. The FTLD therapeutic agent according to claim 1 or 2, wherein, In equation (1-1), n ​​is 2.

4. The FTLD therapeutic agent according to claim 1 or 2, wherein, In equation (1-1), R 1 It is n-propyl.

5. The FTLD therapeutic agent according to claim 3, wherein, In equation (1-1), R 1 It is n-propyl.

6. The FTLD therapeutic agent according to claim 1 or 2, wherein, The compound represented by formula (1-1) is the same as the compound represented by formula (1-2) below: That is, 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one.

7. The FTLD therapeutic agent according to claim 1 or 2, wherein, The pharmaceutically acceptable salt of the compound represented by formula (1-1) is the hydrochloride salt of the compound represented by formula (1-2) below: That is, 4-[2-(dipropylamino)ethyl]-1,3-dihydro-2H-indole-2-one hydrochloride.

8. The FTLD therapeutic agent according to claim 1 or 2, wherein, The compound represented by formula (2-1) is the same as the compound represented by formula (2-2) below: That is, 4-amino-N-(3,4-dimethyl-5-isoxazolyl)benzenesulfonamide.

9. The FTLD therapeutic agent according to claim 1 or 2, wherein, The compound represented by formula (3-1) is the same as the compound represented by formula (3-2) below: That is, 4'-[[4-methyl-6-(1-methyl-1H-benzimidazol-2-yl)-2-propyl1H-benzimidazol-1-yl]methyl]biphenyl-2-carboxylic acid.

10. A pharmaceutical composition for the treatment of frontotemporal degenerative disease (FTLD), comprising the following substances as active ingredients: The following compound is represented by formula (1-1): In equation (1-1), R 1 Each of these can independently represent an alkyl group or a 4-hydroxyphenylethyl group with 1 to 6 carbon atoms, where n represents an integer from 1 to 3. The following compound is represented by formula (2-1): In equation (2-1), R 21 The group consisting of free hydrogen, fatty acid acyl groups with 2 to 18 carbon atoms, and aromatic carboxylic acid acyl groups with 7 to 9 carbon atoms is selected. The following compound is represented by formula (3-1): In equation (3-1), R 31 Choose from the group consisting of hydrogen, halogens, alkyl groups with 1 to 6 carbon atoms, and CF3. R 32 The group consisting of alkoxy groups with 1 to 6 carbon atoms substituted by imidazole groups and optionally substituted nitrogen-containing aromatic heterocycles is selected. R 33 The group consisting of hydrogen, alkyl groups having 1 to 6 carbon atoms, and cycloalkyl groups having 3 to 6 carbon atoms is selected. R 34 Select from the group consisting of carboxyl, cyano, and 1H-tetrazole groups; Their pharmaceutically acceptable salts or their solvates.

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