Use of g923-0271 in the treatment or prevention of tdp-43 associated diseases

Abstract

The application belongs to the technical field of biological medicine, and discloses application of G923-0271 in treatment or prevention of TDP-43 related diseases, and specifically discloses application of G923-0271 or a derivative thereof in inhibition of TDP-43 expression or preparation of a TDP-43 inhibitor. The inventors have found that G923-0271 can reduce the protein level of overexpressed human TDP-43 at the cell level, and meanwhile, G923-0271 can improve the disease phenotype of a TDP-43 nematode model by reducing the level of TDP-43, indicating that G923-0271 can be used for treating diseases caused by abnormal expression of TDP-43 protein, such as ALS and FTD and the like, and is expected to be developed into a new anti-TDP-43 drug.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of G923-0271 in the treatment or prevention of TDP-43 related diseases. Background Technology

[0002] TARDNA-binding protein 43 (TDP-43) is a highly conserved nuclear RNA / DNA binding protein encoded by the TARDBP gene. It consists of 414 amino acids and belongs to the heterologous nucleoribonucleoprotein (hnRNP) family. Normally, TDP-43 is mainly found in the cell nucleus, playing roles in transcriptional regulation, alternative splicing, and mRNA stabilization. Under pathological conditions, TDP-43 may undergo splicing, hyperphosphorylation, and ubiquitination, leading to its accumulation and aggregation in the cytoplasm. Combined with its prion-like domain, this induces misfolding and self-aggregation, causing intercellular spread and accelerating cytotoxicity. Furthermore, TDP-43 can participate in liquid-liquid phase separation; mutations or abnormal post-translational modifications can lead to irreversible aggregation of TDP-43 through liquid-solid phase separation, resulting in cytotoxicity. The accumulation of TDP-43 aggregates in the central nervous system is a common feature of diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Reducing misfolded TDP-43 and TDP-43 aggregates will guide new treatment strategies for TDP-43-related diseases.

[0003] The compound 3-(4-fluorophenyl)-6-(4-methylphenyl)-N-[(4-methylphenyl)methyl]-2H-pyrazolo[3,4-b]pyridine-4-carboxamide (PubChem CID: 16033417, https: / / pubchem.ncbi.nlm.nih.gov / compound / 16033417), has the molecular formula C 28 H 23 FN4O, with a relative molecular weight of 450.5 g / mol, CAS: 959554-92-8, has the following structural formula: Figure 1 As shown in Figure B, this compound is referred to as G923-0271 in this patent. As a small molecule compound, G923-0271 has not been clearly reported in the field of drug preparation for inhibiting TDP-43 protein in TDP-43-related diseases. Summary of the Invention

[0004] The first aspect of the present invention is to provide the use of G923-0271 or its derivatives in inhibiting TDP-43 expression or in the preparation of TDP-43 inhibitors.

[0005] A second aspect of the present invention aims to provide the use of G923-0271 or its derivatives in the preparation of medicaments for treating, improving and / or preventing diseases.

[0006] A third aspect of the present invention aims to provide the use of G923-0271 or its derivatives in the preparation of a medicament for improving the loss of motor neurons in animals.

[0007] The fourth aspect of this invention is to provide a product.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0009] A first aspect of the present invention provides the use of G923-0271 or a derivative thereof in inhibiting TDP-43 expression or in the preparation of TDP-43 inhibitors, wherein G923-0271 is 3-(4-fluorophenyl)-6-(4-methylphenyl)-N-[(4-methylphenyl)methyl]-2H-pyrazolo[3,4-b]pyridine-4-carboxamide.

[0010] In some embodiments of the present invention, the G923-0271 or its derivatives can inhibit the expression of TDP-43 protein.

[0011] In some embodiments of the present invention, the derivatives include pharmaceutically acceptable salts, hydrates, solvent compounds, polymorphs, tautomers, and prodrugs.

[0012] In some embodiments of the present invention, the pharmaceutically acceptable salt includes at least one of hydrochloride, hydrobromide, acetate, citrate, benzoate, succinate, octanoate, fumarate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, tartrate, maleate, or trifluoroacetate.

[0013] In some embodiments of the present invention, the effective concentration of G923-0271 or its derivatives is 1 to 100 μM.

[0014] In some preferred embodiments of the present invention, the effective concentration of G923-0271 or its derivative is 5 to 100 μM.

[0015] A second aspect of the invention provides the use of G923-0271 or a derivative thereof in the preparation of medicaments for treating, improving, and / or preventing diseases, including diseases caused by abnormal TDP-43 expression, wherein G923-0271 is 3-(4-fluorophenyl)-6-(4-methylphenyl)-N-[(4-methylphenyl)methyl]-2H-pyrazolo[3,4-b]pyridine-4-carboxamide.

[0016] In some embodiments of the present invention, the drug achieves the purpose of treating, improving and / or preventing diseases by inhibiting the expression of TDP-43.

[0017] In some embodiments of the present invention, the disease includes at least one of amyotrophic lateral sclerosis, frontotemporal dementia, and Alzheimer's disease.

[0018] In some embodiments of the present invention, the derivatives include pharmaceutically acceptable salts, hydrates, solvent compounds, polymorphs, tautomers, and prodrugs.

[0019] In some embodiments of the present invention, the pharmaceutically acceptable salt includes at least one of hydrochloride, hydrobromide, acetate, citrate, benzoate, succinate, octanoate, fumarate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, tartrate, maleate, or trifluoroacetate.

[0020] In some embodiments of the present invention, the drug further comprises one or more pharmaceutically acceptable excipients.

[0021] In some embodiments of the present invention, the pharmaceutically acceptable excipients comprise at least one of diluents, binders, wetting agents, lubricants, disintegrants, solvents, emulsifiers, cosolvents, solubilizers, preservatives, pH adjusters, osmotic pressure adjusters, surfactants, coating materials, antioxidants, antibacterial agents, or buffers.

[0022] In some embodiments of the present invention, the dosage form of the drug includes at least one of solid dosage form, liquid dosage form and semi-solid dosage form.

[0023] In some embodiments of the present invention, the drug comprises at least one of tablets, capsules, granules, pills, drop pills, liquid preparations, decoctions, suspensions, dispersants, syrups, suppositories, gels, aerosols, and patches.

[0024] In some embodiments of the present invention, the route of administration of the drug includes at least one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration, nebulized administration, or transdermal administration.

[0025] In some embodiments of the present invention, the effective concentration of G923-0271 or its derivatives is 25-100 μM.

[0026] In some preferred embodiments of the present invention, the effective concentration of G923-0271 or its derivative is 50-100 μM.

[0027] In some preferred embodiments of the present invention, the effective concentration of G923-0271 or its derivative is 60-100 μM.

[0028] A third aspect of the invention provides the use of G923-0271 or a derivative thereof in the preparation of a medicament for improving the loss of motor neurons in animals, wherein G923-0271 is 3-(4-fluorophenyl)-6-(4-methylphenyl)-N-[(4-methylphenyl)methyl]-2H-pyrazolo[3,4-b]pyridine-4-carboxamide.

[0029] In some embodiments of the present invention, the animal includes nematodes.

[0030] In some embodiments of the present invention, the derivatives include pharmaceutically acceptable salts, hydrates, solvent compounds, polymorphs, tautomers, and prodrugs.

[0031] In some embodiments of the present invention, the pharmaceutically acceptable salt includes at least one of hydrochloride, hydrobromide, acetate, citrate, benzoate, succinate, octanoate, fumarate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, tartrate, maleate, or trifluoroacetate.

[0032] In some embodiments of the present invention, the effective concentration of G923-0271 or its derivatives is 25-100 μM.

[0033] In some preferred embodiments of the present invention, the effective concentration of G923-0271 or its derivative is 50-100 μM.

[0034] In some preferred embodiments of the present invention, the effective concentration of G923-0271 or its derivative is 60-100 μM.

[0035] In some embodiments of the present invention, the drug further comprises one or more pharmaceutically acceptable excipients.

[0036] In some embodiments of the present invention, the pharmaceutically acceptable excipients comprise at least one of diluents, binders, wetting agents, lubricants, disintegrants, solvents, emulsifiers, cosolvents, solubilizers, preservatives, pH adjusters, osmotic pressure adjusters, surfactants, coating materials, antioxidants, antibacterial agents, or buffers.

[0037] In some embodiments of the present invention, the dosage form of the drug includes at least one of solid dosage form, liquid dosage form and semi-solid dosage form.

[0038] In some embodiments of the present invention, the drug comprises at least one of tablets, capsules, granules, pills, drop pills, liquid preparations, decoctions, suspensions, dispersants, syrups, suppositories, gels, aerosols, and patches.

[0039] In some embodiments of the present invention, the route of administration of the drug includes at least one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration, nebulized administration, or transdermal administration.

[0040] A fourth aspect of the present invention provides a product comprising G923-0271 or a derivative thereof, wherein G923-0271 is 3-(4-fluorophenyl)-6-(4-methylphenyl)-N-[(4-methylphenyl)methyl]-2H-pyrazolo[3,4-b]pyridine-4-carboxamide.

[0041] In some embodiments of the present invention, the effective concentration of G923-0271 or its derivatives is 1 to 100 μM.

[0042] In some preferred embodiments of the present invention, the effective concentration of G923-0271 or its derivative is 5 to 100 μM.

[0043] In some embodiments of the present invention, the product has any of the following functions:

[0044] (1) Inhibit TDP-43 expression;

[0045] (2) To treat, improve and / or prevent diseases caused by abnormal TDP-43 expression;

[0046] (3) Improves the loss of motor neurons in animals.

[0047] In some embodiments of the present invention, the product is a drug or reagent.

[0048] In some embodiments of the present invention, the drug further comprises pharmaceutically acceptable excipients.

[0049] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one of the following: diluents, binders, wetting agents, lubricants, disintegrants, solvents, emulsifiers, cosolvents, solubilizers, preservatives, pH adjusters, osmotic pressure adjusters, surfactants, coating materials, antioxidants, antibacterial agents, or buffers.

[0050] In some embodiments of the present invention, the dosage form of the drug includes at least one of suspension, granules, capsules, powders, tablets, emulsions, solutions, pellets, injections, oral preparations, suppositories, enemas, aerosols, patches, or drops.

[0051] In some embodiments of the present invention, the route of administration of the drug includes at least one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration, nebulized administration, or transdermal administration.

[0052] The beneficial effects of this invention are:

[0053] The inventors discovered that G923-0271 can reduce the protein level of overexpressed human TDP-43 at the cellular level. At the same time, G923-0271 can improve the disease phenotype of TDP-43 nematode models by reducing TDP-43 levels (significantly improving paralysis, prolonging lifespan, improving behavioral disorders, and reducing motor neuron loss in TDP-43 nematodes). This suggests that G923-0271 can be used to treat diseases caused by abnormal TDP-43 protein expression, such as ALS and FTD, and is expected to be developed into a novel anti-TDP-43 drug. Attached Figure Description

[0054] Figure 1 The expression of exogenous TDP-43 protein after treating HeLa EGFP WT-TDP-43 and HeLa EGFP A315T-TDP-43 cell lines with G923-0271; where A is the result of Western blot detection; B is the chemical structure diagram of G923-0271, and * in the figure represents p < 0.05.

[0055] Figure 2G923-0271 alleviated the disease phenotype of TDP-43 transgenic nematodes by reducing TDP-43 protein levels. Specifically, A shows the effect of G923-0271 on the growth and development of N2 nematodes; B is a schematic diagram of the construction of unc::47::TDP-43-A315T transgenic nematodes; C shows the effect of G923-0271 on paralysis in TDP-43 transgenic nematodes, significantly improving paralysis; D shows the effect of G923-0271 on the lifespan of TDP-43 transgenic nematodes, significantly prolonging their lifespan; E shows the TDP-43 protein level detection results in TDP-43 transgenic nematodes; and F shows the TDP-43 protein level... Statistical results of TDP-43 protein levels in transgenic nematodes: G represents the effect of G923-0271 on behavioral disorders in TDP-43 transgenic nematodes, which can significantly improve behavioral disorders in TDP-43 transgenic nematodes; H is a schematic diagram of hybridization between unc::47::TDP-43-A315T transgenic nematodes and unc::47::GFP transgenic nematodes; I is a real graph showing the improvement of motor neuron loss in TDP-43 transgenic nematodes by G923-0271; J is a statistical graph showing the results of G923-0271 in improving motor neuron loss in TDP-43 transgenic nematodes. In the figure, * represents p < 0.05, ** represents p < 0.01, and *** represents p < 0.005. Detailed Implementation

[0056] The present invention will be further described in detail below through specific embodiments.

[0057] It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0058] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0059] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0060] Example 1: Effect of small molecule compound G923-0271 on the expression of endogenous and exogenous TDP-43 in cells.

[0061] Construction of HeLa EGFP WT-TDP-43 cells and HeLa EGFP A315T-TDP-43 plasmid: The vector plasmid FUGW-UBC-P2A-EGFP was linearized by double digestion with restriction endonucleases NheI and BamHI. The human TDP-43 insert (WT and A315T mutants) was amplified by PCR using recombinant primers (TDP43-Asb-F1:5'-GCAGGTCGACTCTAGAGGATCGGCTAGCGCCACCATGGACTACAAAGACCATGACG-3' (SEQ ID NO:1); TDP43-Asb-R1:5'-GCAGAGAGAAGTTTGTGGCGCCGGATCCCATTCCCCAGCCAGAAGACTTAGAATCC-3' (SEQ ID NO:2)). After DNA gel electrophoresis of the digestion products and PCR products, the linearized FUGW-UBC-P2A-EGFP and TDP43 fragments were recovered using a DNA fragment recovery kit. Using the Gibson Assembly kit, the linearized FUGW-UBC-P2A-EGFP7 and TDP43 fragments were assembled into a recombinant plasmid FUGW-UBC-TDP43-P2A-EGFP. After verification by enzyme digestion and Sanger sequencing, the plasmid was confirmed to be endotoxin-free and stored at -20°C for later use.

[0062] Construction of HeLa EGFP WT-TDP-43 and HeLa EGFP A315T-TDP-43 cells: First, Flag-TDP-43-WT-EGFP and Flag-TDP-43-A315T-EGFP were packaged into lentiviruses for 293TN cells using the Lentiviral III packaging system. HeLa cells were then transfected with the lentiviral particles. 72 hours after transfection, the culture medium was replaced with fresh antibiotic-containing medium. After one week of selection, single clones were picked and amplified. The resulting cell lines were verified by Western blotting, immunofluorescence, and sequencing, successfully constructing stably expressing HeLa EGFP WT-TDP-43 and HeLa EGFP A315T-TDP-43 cells.

[0063] The constructed HeLa EGFP WT-TDP-43 and HeLa EGFP A315T-TDP-43 cells were added to DMEM medium containing 10 v / v % FBS and cultured at 37°C under 5% CO2 conditions. Before drug treatment, the cells were digested into single cells by treating with 0.05% trypsin for 3 minutes and then cultured at 5 × 10⁻⁶ cells / year. 5Cells were seeded into 6-well plates at the specified density and cultured for 24 hours. HeLa EGFP WT-TDP-43 and HeLa EGFP A315T-TDP-43 cells were then treated for 24 hours with 1 μM, 5 μM, and 10 μM G923-0271 (10 mM stock solution dissolved in DMSO, diluted to the final concentration in culture medium), respectively. A control group was also included (treated with the same volume of DMSO for 24 hours). Cells were then lysed with RIPA strong lysis buffer for 30 minutes, sonicated every 10 minutes, and centrifuged at 15000g for 30 minutes to obtain total cellular protein. Cellular protein concentration was determined using the BCA assay, and TDP-43 protein expression was detected by Western blotting, using β-actin as an internal control. Flag antibody was used to indicate the content of exogenous TDP-43 protein.

[0064] The results are as follows Figure 1 As shown in Figure A, treatment of HeLa EGFP WT-TDP-43 and HeLaEGFP A315T-TDP-43 cells with 5 μM and 10 μM G923-0271 for 24 hours significantly reduced the expression level of exogenous TDP-43 protein (p < 0.05).

[0065] Example 2: Effects of G923-0271 on the growth and development of N2 nematodes

[0066] Preparation of NGM medium for nematodes: Weigh 3 g sodium chloride, 17 g agar, and 2.5 g peptone into 975 mL of water. After sterilization in an autoclave, add 1 mL of 1 mol calcium chloride solution, 1 mL of 1 mol magnesium sulfate solution, 1 mL of 5 mg / mL cholesterol, and 25 mL of 1 mol potassium phosphate.

[0067] Preparation of drug culture plates: Dilute 50 mM G923-0271 drug to final concentrations of 10, 50, 100, and 150 μM using NGM medium, pour the solution into culture plates, and store at 4°C in the dark after solidification to obtain the drug culture plates.

[0068] On the first day, adult N2 nematodes were transferred to a drug culture plate to lay eggs. Two hours later, the adults were removed, and the number of eggs laid by the nematodes was counted under a microscope. Then, the growth of the nematodes was observed for three consecutive days, including the number of nematodes from eggs to L1 stage, the number of nematodes that grew to L4 stage, and the number of adult nematodes.

[0069] The results showed that after treatment with different concentrations (0, 10, 50, 100, 150 μM) of G923-0271, there were no significant differences in the number of eggs laid, the number of eggs reaching the L1 stage, the number of nematodes reaching the L4 stage, and the number of adult nematodes in N2 nematodes, indicating that G923-0271 had no effect on the growth and development of N2 nematodes. Figure 2 (A) indicates that G923-0271 has a high level of security.

[0070] Example 3: Effects of G923-0271 on the disease phenotype of TDP-43 transgenic nematode

[0071] Construction of plasmid unc47::TDP-43-A315T: The vector plasmid unc47-sl2-DsRed2 (gifted by Zhejiang University) was double-digested with restriction endonuclease XbaI and EcoRI. During linearization, the sl2-DsRed2 sequence was removed, retaining only the unc47 promoter sequence. The human TDP-43 insert was amplified by PCR using recombinant primers (TDP43-Asb-F1:5'-GTCACATTTATTTCATTACAGGGATCCCTGCTCTAGAGGTGGAGGTTCAATGTCTGAATATATTCGG-3' (SEQ ID NO:3); TDP43-Asb-R1:5'-GCGACCGGCGCTCGAGTTAGAATTCTACATTCCCCAGCCAGAAGACTTAGAATCC-3' (SEQ ID NO:4)). After DNA gel electrophoresis of the enzyme digestion products and PCR products, the linearized unc47 and TDP-43 fragments were recovered using a DNA fragment recovery kit. The linearized unc47 and TDP-43 fragments were then assembled into a recombinant plasmid unc47::TDP-43 using a Gibson Assembly kit. After verification by enzyme digestion and Sanger sequencing, the endotoxin-free plasmid was extracted and stored at -20°C for later use.

[0072] Paralysis test: Thirty adult nematodes on day 1 were placed on an NGM board, and their movement was observed daily. If a nematode remained motionless after being poked with a nematode pick on the NGM board, but could still feed, it was considered paralyzed.

[0073] Lifespan determination: To determine the lifespan of nematodes, 30 nematodes on an NGM plate were maintained at 20°C from the first day of adulthood until death. Nematodes that did not respond to touch and were unable to feed were declared dead.

[0074] Nematode protein extraction: First-day adult nematodes were washed off with M9 buffer and transferred to 1.5 mL EP tubes. The tubes were placed on ice and allowed to precipitate after 10 minutes. The nematodes were then washed again with M9 buffer, repeated three times. The supernatant was thoroughly aspirated, and the tubes were stored at -80°C overnight. The nematodes were then lysed using RIPA lysis buffer for 30 minutes, sonicated every 10 minutes, centrifuged, and the supernatant was collected as total nematode protein. TDP-43 protein expression was detected by Western blotting. The content of exogenous TDP-43 protein was indicated using a TDP-43 antibody.

[0075] Nematode swimming test: Thirty adult nematodes on day 1 were placed in one drop of M9 buffer. After the nematodes adapted to the M9 buffer for one minute, the nematodes' waving was recorded as a video for one minute.

[0076] Preparation of TDP-43 transgenic nematodes for visual motor neurons: The unc47::TDP-43-A315T transgenic nematodes were hybridized with the unc47::GFP transgenic nematodes. After self-pollination, the resulting nematodes were selected as dihozygous transgenic nematodes. The specific steps of nematode hybridization included: (1) inducing male nematodes to be generated by unc47::GFP nematodes at 30℃; (2) 3 male unc47::GFP nematodes were hybridized with hermaphroditic unc47::TDP-43-A315T transgenic nematodes (obtained by Fujian Shangyuan Biotechnology Co., Ltd. through microinjection and ultraviolet integration of the unc47::TDP-43-A315T plasmid into the N2 nematode genome for stable expression). (15) were placed in the same NGM culture dish and allowed to mate freely; (3) F1 female nematodes with GFP fluorescence were selected and allowed to self-fertilize to produce F2 generation. 20 nematodes with the strongest fluorescence intensity (unc47::GFP homozygote) were selected and placed in separate NGM dishes to continue self-fertilizing to produce F3 generation. 10 larvae from each F3 generation were selected for PCR identification. All 10 larvae were TDP-43 positive, confirming that the F2 generation nematodes were suspected to be TDP-43 homozygotes; (4) The suspected TDP-43 homozygotes were self-fertilized and PCR identification was continued. All 3 generations of nematodes were TDP-43 positive, indicating that the unc47::TDP-43-A315T::GFP double transgenic nematodes).

[0077] Statistical analysis of nematode motor neurons: Imaging was performed using a laser confocal microscope (nematode slide preparation: 1% agar was prepared, heated, and a drop was placed in the center of a slide. A coverslip was then placed over the slide, and after solidification, the coverslip was removed. 2.5 μL of 10 μM levamisole was added to the agar to anesthetize the nematodes. Then, up to 5 nematodes were placed in the levamisole solution, and the coverslip was placed over the slide for imaging). This invention revealed that 75 μM G923-0271 significantly inhibited the loss of motor neurons in diseased nematodes on day 5 of nematode maturation.

[0078] The culture conditions and methods for all nematodes were the same as in Example 2.

[0079] The transgenic nematode unc47::TDP-43-A315T was stably expressed by transferring the TDP-43-A315T plasmid into the genome of N2 nematode through microinjection and ultraviolet integration by Fujian Shangyuan Biotechnology Co., Ltd. This invention obtained a TDP-43 transgenic model nematode containing the A315T mutation after backcrossing the unc47::TDP-43-A315T transgenic nematode with N2 nematode for four generations, thus removing the background. Figure 2 (B) Transgenic nematodes of N2 and unc47::TDP-43-A315T were cultured at 20°C. Under 50 μM and 75 μM conditions, G923-0271 significantly improved the paralysis rate of the transgenic nematodes of unc47::TDP-43-A315T. Figure 2 (C), of which 75 μM G923-0271 treatment can significantly prolong the survival of diseased nematodes (C). Figure 2 (D). Subsequent experimental results demonstrated that treatment with 75 μM G923-0271 significantly reduced the exogenous TDP-43 content in transgenic nematodes of unc47::TDP-43-A315T ( Figure 2 (E-F) and significantly improved the swimming impairment of nematodes ( Figure 2 (G). By hybridizing unc47::TDP-43-A315T transgenic nematodes and unc47::GFP transgenic nematodes, a double transgenic nematode with visual motor neurons was obtained (G). Figure 2 Treatment with the drug G923-0271 significantly delayed the loss of motor neurons in diseased nematodes. Figure 2 (I~J).

[0080] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. The use of G923-0271 or a pharmaceutically acceptable salt thereof in the preparation of medicaments for treating, improving, and / or preventing diseases, said diseases being caused by abnormal TDP-43 expression, wherein G923-0271 is 3-(4-fluorophenyl)-6-(4-methylphenyl)-N-[(4-methylphenyl)methyl]-2H-pyrazolo[3,4-b]pyridine-4-carboxamide; said diseases being at least one of amyotrophic lateral sclerosis, frontotemporal dementia, and Alzheimer's disease.

2. The application according to claim 1, characterized in that, The effective concentration of G923-0271 or its pharmaceutically acceptable salt is 25~100 μM.

3. A drug having G923-0271 or a pharmaceutically acceptable salt thereof as its active ingredient, wherein G923-0271 is 3-(4-fluorophenyl)-6-(4-methylphenyl)-N-[(4-methylphenyl)methyl]-2H-pyrazolo[3,4-b]pyridine-4-carboxamide; wherein the effective concentration of G923-0271 or a pharmaceutically acceptable salt thereof is 1-100 μM.

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