Oxadiazole derivative compounds and pharmaceutical compositions containing the same

Abstract

The present invention relates to a novel compound having histone deacetylase 6 (HDAC6) inhibitory activity, its stereoisomers, its pharmaceutically acceptable salts, their uses for the manufacture of pharmaceuticals, pharmaceutical compositions and methods for prevention or treatment containing them, and a method for producing a novel 1,3,4-oxadiazole oxetane derivative.

Description

[Technical Field]

[0001] The present invention relates to a novel oxadiazole derivative compound, its stereoisomer or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the oxadiazole derivative compound, its stereoisomer or a pharmaceutically acceptable salt thereof, uses of the oxadiazole derivative compound, its stereoisomer or a pharmaceutically acceptable salt thereof for the manufacture of therapeutic agents, a method for preventing or treating a disease by administering the oxadiazole derivative compound, its stereoisomer or a pharmaceutically acceptable salt thereof, and a method for producing the oxadiazole derivative compound, its stereoisomer or a pharmaceutically acceptable salt thereof. [Background technology]

[0002] Post-translational modifications, such as acetylation, within cells are crucial regulatory modules that play a central role in biological processes and are strictly controlled by numerous enzymes. Histones are core proteins that make up chromatin; they act as axes around which DNA wraps and promote DNA condensation. Furthermore, the balance between histone acetylation and deacetylation plays a very important role in gene expression.

[0003] Histone deacetylases (HDACs) are enzymes that remove acetyl groups from lysine residues in histone proteins that make up chromatin. They are involved in gene silencing and are known to induce cell cycle arrest, inhibition of angiogenesis, immunomodulation, and cell death (Hassig et al., Curr. Opin. Chem. Biol. 1997, 1,300-308). Furthermore, it has been reported that inhibition of HDAC enzyme function degrades the activity of cancer cell survival-related factors and activates cancer cell death-related factors, thereby inducing apoptosis in cancer cells (Warrell et al, J. Natl. Cancer Inst. 1998, 90, 1621-1625).

[0004] In humans, 18 HDACs are known and are classified into four classes according to their homology with yeast HDACs. Of these, 11 HDACs that use zinc as a cofactor can be classified into three groups: Class I (HDAC1, 2, 3, 8), Class II (IIa: HDAC4, 5, 7, 9; IIb: HDAC6, 10), and Class IV (HDAC11). Furthermore, seven HDACs in Class III (SIRT1-7) use NAD+ as a cofactor instead of zinc (Bolden et al., Nat. Rev. Drug. Discov. 2006, 5(9), 769-784).

[0005] While various HDAC inhibitors are in the preclinical or clinical development stages, only non-selective HDAC inhibitors have been known as anticancer drugs until now. Vorinostat (SAHA) and romidepsin (FK228) are approved for the treatment of cutaneous T-cell lymphoma, and panobinostat (LBH-589) is approved for the treatment of multiple myeloma. However, non-selective HDAC inhibitors are generally known to cause side effects such as fatigue and nausea at high doses (Piekarz et al., Pharmaceuticals 2010, 3, 2751-2). 767). Such side effects have been reported to be caused by Class I HDAC inhibition, and due to these side effects, non-selective HDAC inhibitors have faced limitations in drug development in fields other than anticancer drugs (Witt et al., Cancer Letters 277 (2009) 8.21).

[0006] On the other hand, there are reports that selective Class II HDAC inhibition does not exhibit the toxicity associated with Class I HDAC inhibition. Therefore, if selective HDAC inhibitors are developed, the side effects such as toxicity caused by non-selective HDAC inhibition can be resolved, and selective HDAC inhibitors have the potential to be developed as effective treatments for various diseases (Matthias et al., Mol.Cell.Biol.2008,28,1688-1701).

[0007] HDAC6, a Class IIb HDAC, is primarily found in the cytoplasm and is known to be involved in the deacetylation of numerous non-histone substrates (such as HSP90 and cortactin), including tubulin proteins (Yao et al., Mol. Cell 2005, 18, 601-607). HDAC6 has two catalytic domains, and its C-terminal zinc finger domain can bind to ubiquitinated proteins. Because HDAC6 has many non-histone proteins as substrates, it is known to play an important role in various diseases, including cancer, inflammatory diseases, autoimmune diseases, neurological diseases, and neurodegenerative disorders. (Santo et al., Blood 2012 119:2579-2589; Vishwakarma et al., International Immunopharmacology 2013, 16, 72-78; Hu et al., J. Neurol. Sci. 2011, 304, 1-8).

[0008] The common structural features of various HDAC inhibitors, as shown in the structure of vorinostat below, consist of a cap group, a linker group, and a zinc binding group (ZBG). Many researchers have studied the inhibitory activity and selectivity of enzymes through structural modifications of the cap group and linker group. Of these, the zinc binding group is known to play a more important role in enzyme inhibitory activity and selectivity (Wiest et al., J.Org.Chem.2013 78:5051-5055; Methot et al., Bioorg.Med.Chem.Lett.2008,18,973-978).

[0009] [Chemical formula]

[0010] Most of the zinc-binding groups are hydroxamic acid or benzamide. Among them, hydroxamic acid derivatives show strong HDAC inhibitory effects, but have problems of low bioavailability and severe off-target activity. In the case of benzamide, since it contains aniline, there is a problem that toxic metabolites can be generated in vivo (Woster et al., Med. Chem. Commun. 2015, online publication).

[0011] Therefore, for the treatment of cancer, inflammatory diseases, autoimmune diseases, neurological diseases, neurodegenerative disorders, etc., different from non-selective inhibitors with side effects, there is a need to develop selective HDAC6 inhibitors with improved bioavailability and no side effects having a zinc-binding group.

Prior Art Documents

Patent Documents

[0012]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

[0013] The object of the present invention is to provide oxadiazole derivative compounds having selective HDAC6 inhibitory activity, stereoisomers thereof, or pharmaceutically acceptable salts thereof.

[0014] Another object of the present invention is to provide a pharmaceutical composition comprising an oxadiazole derivative compound having selective HDAC6 inhibitory activity, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0015] Another object of the present invention is to provide a method for producing oxadiazole derivative compounds, stereoisomers thereof, or pharmaceutically acceptable salts thereof.

[0016] Another object of the present invention is to provide a pharmaceutical composition comprising an oxadiazole derivative compound, its stereoisomer, or a pharmaceutically acceptable salt thereof for the prevention or treatment of diseases or degenerative brain diseases associated with HDAC6 activity.

[0017] Another object of the present invention is to provide uses for oxadiazole derivative compounds, stereoisomers thereof, or pharmaceutically acceptable salts thereof for the manufacture of agents for the prevention or treatment of diseases or degenerative brain diseases associated with HDAC6 activity.

[0018] Another object of the present invention is to administer therapeutically effective amounts of oxadiazole derivative compounds, their stereoisomers, or pharmaceutically acceptable salts thereof, for diseases related to the activity of HDAC6. Alternatively, it may involve providing methods for preventing or treating degenerative brain diseases.

[0019] Another object of the present invention is to provide applications for the prevention or treatment of diseases or degenerative brain diseases related to the activity of HDAC6 in oxadiazole derivative compounds, their stereoisomers, or pharmaceutically acceptable salts thereof. [Means for solving the problem]

[0020] The inventors of this invention have discovered an oxadiazole derivative compound having histone deacetylase 6 (HDAC6) inhibitory activity, and have completed the present invention by using this compound to inhibit or treat diseases related to HDAC6 activity.

[0021] This will be explained in detail below. All combinations of the various elements disclosed in this invention fall within the scope of the invention. Furthermore, the scope of the invention is not limited by the following detailed explanation.

[0022] compound The present invention provides a compound relating to any one of the following (1) to (7), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0023] (1) Compounds represented by the following chemical formula I, their stereoisomers, or pharmaceutically acceptable salts: [ka] In the above chemical formula I, X1 to X4 are each independently N or CR , , a , , , x , , ,

[0025] , , , a , , a , , x , , wherein, X1 to X4 cannot be N at the same time for three or more, and R x is -H, F, Cl, Br or I; R1 is -CX a H2, -C(X a )2H, or -C(X a )3, wherein Xa is F, Cl, Br or I; R2 and R3 are each independently F, Cl, Br or I.

[0024] (2) A compound represented by the chemical formula I according to (1) above, its stereoisomer or its pharmaceutically acceptable salt: X1 to X4 are each independently N or -CR x wherein, X1 to X4 cannot be N at the same time for three or more, and R x is H or F; R1 is -CX a H2 or -C(X a )2H, wherein X a is F or Cl; R2 and R3 may each independently be F or Cl.

[0025] (3) A compound represented by the chemical formula I according to (1) or (2) above, its stereoisomer or its pharmaceutically acceptable salt: The compound represented by the above chemical formula I may be a compound represented by the following chemical formula II. [Chemical formula] <s In the above chemical formula II, X2 is N or CR x wherein, R x is H, F, Cl, Br or I; R1 is -CX a H2, -C(X a )2H, or -C(X a)3, where Xa is F, Cl, Br or I; R2 and R3 may each be independently F, Cl, Br, or I.

[0026] (4) Compounds represented by chemical formula I according to (1), (2) or (3) above, stereoisomers thereof, or pharmaceutically acceptable salts thereof: The compound represented by chemical formula I or chemical formula II may also be a compound represented by the following chemical formulas II-1, II-2, II-3, or II-4.

[0027] [ka]

[0028] [ka]

[0029] [ka]

[0030] [ka]

[0031] In the aforementioned chemical formulas II-1, II-2, II-3, or II-4, X2 is either N or -CR in each chemical formula independently. x And R x is H, F or Cl, Br or I; R1 is independently -CX in each chemical formula. a H2, -C(X a )2H, or -C(X a )3, where Xa is independently F, Cl, Br, or I in each chemical formula; R2 and R3 are, independently, F, Cl, Br, or I in their respective chemical formulas.

[0032] (5) Compounds represented by chemical formula I according to (1), (2), (3) or (4) above, Stereoisomers of or pharmaceutically acceptable salts thereof: In the aforementioned chemical formulas II-1, II-2, II-3, or II-4, X2 is either N or -CR in each chemical formula independently. x And here, R x is either H or F; R1 is independently -CX in each chemical formula. a H2 or -C(X a )3, where X a is either F or Cl; R2 and R3 are independently F or Cl in their respective chemical formulas.

[0033] In embodiments of the present invention, R2 and R3 in chemical formulas II-1, II-2, II-3, or II-4 may be the same or different from each other.

[0034] In embodiments of the present invention, in chemical formula I, chemical formula II, or chemical formula I-1, R2 and R3 may be the same or different from each other, and in particular, R2 and R3 may be different from each other. For example, if one of R2 and R3 is F, the other may be Cl.

[0035] In embodiments of the present invention, in chemical formulas I, II, II-2, II-3, or II-4, R2 and R3 may be the same or different from each other. In particular, R2 and R3 may be the same from each other. For example, both R2 and R3 may be F, or both may be Cl.

[0036] (6) Compounds represented by chemical formula I according to (1), (2), (3), (4) or (5) above, stereoisomers thereof, or pharmaceutically acceptable salts thereof: The compound represented by chemical formula I may be any one selected from compounds 1 to 8 listed in Table 1 below. [Table 1]

[0037] (7) Compounds represented by chemical formula I according to (1), (2), (3), (4), (5) or (6) above, stereoisomers thereof, or pharmaceutically acceptable salts thereof: The compound represented by chemical formula I may be compound 1 or compound 5 as listed in Table 1.

[0038] In this invention, the term "pharmaceutically acceptable" may also mean physiologically acceptable and, when administered to a subject, not typically cause gastrointestinal disorders, dizziness, or other allergic or similar reactions. The pharmaceutically acceptable salts of the present invention may be produced by conventional methods known to those skilled in the art.

[0039] In this invention, pharmaceutically acceptable salts mean salts commonly used in the pharmaceutical industry, such as inorganic ion salts produced from calcium, potassium, sodium, or magnesium; inorganic acid salts produced from hydrochloric acid, nitric acid, phosphoric acid, bromate, iodic acid, perchloric acid, or sulfuric acid; acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carboxylic acid, vanillic acid, hydroiodic acid, organic acid salts produced from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or naphthalenesulfonic acid; amino acid salts produced from glycine, arginine, lysine, and other sources; and trimethylamine, triethylamine, ammonia. Examples include amine salts produced from nia, pyridine, picoline, etc., but the types of salts referred to in this invention are not limited by these salts.

[0040] Preferred salts in the present invention include hydrochloric acid, trifluoroacetic acid, citric acid, bromate, maleic acid, phosphoric acid, sulfuric acid, and tartaric acid.

[0041] The compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4 of the present invention may contain one or more chiral carbons, thereby existing as racemates, racemic mixtures, single enantiomers (optical isomers), diastereomer mixtures, and their respective diastereomers.

[0042] Such isomers can be separated by conventional techniques, for example, by resolution such as column chromatography or HPLC, for compounds represented by chemical formulas I, II, II-1, II-2, II-3, or II-4. Alternatively, the stereoisomers of each compound represented by chemical formulas I, II, II-1, II-2, II-3, or II-4 can be synthesized stereospecifically using optically pure starting materials and / or reagents having known sequences.

[0043] In the present invention, "stereoisomer" includes diastereomers and enantiomers, and optical isomers include not only enantiomers but also mixtures of enantiomers and racemates.

[0044] The compounds represented by chemical formulas I, II, II-1, II-2, II-3, or II-4 of the present invention, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts exhibit inhibitory activity against histone deacetylase 6 (HDAC6). Specifically, they exhibit selective inhibitory activity against HDAC6, and more specifically, they exhibit very high selective inhibitory activity against HDAC6 compared to other HDAC isotypes. They have very high inhibitory activity against HDAC6, while showing little to no inhibitory activity against other HDAC isotypes (Tables 3, 4, and 5).

[0045] The compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4 of the present invention, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof can significantly increase tubulin acetylation in nerve cells (Figures 4 and 5).

[0046] The compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4 of the present invention, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof can exhibit activity to prevent or treat diseases or diseases mediated by histone deacetylase 6 (HDAC6) activity.

[0047] In the present invention, "prevention" means any action that suppresses or delays the onset of a disease by administering a compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4 of the present invention, a compound represented by compound 1 to 8 listed in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0048] In this invention, "treatment" refers to the compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4 of this invention, or the compounds listed in Table 1. This refers to any action by which the symptoms of a suspected or affected subject are improved or favorably altered by administering a compound represented by 1-8, its stereoisomer, or a pharmaceutically acceptable salt thereof.

[0049] In this specification, "histone deacetylase-mediated disease" means a disease related to the activity of histone deacetylase 6 (HDAC6).

[0050] In embodiments of the present invention, diseases related to histone deacetylase 6 (HDAC6) activity or histone deacetylase 6-mediated diseases may include infectious diseases, neoplasms, endocrine, nutritional and metabolic disorders, mental and behavioral disorders, neurological disorders, ocular and adnexal disorders, cardiovascular diseases, respiratory diseases, digestive diseases, skin and subcutaneous tissue disorders, musculoskeletal and connective tissue disorders, or congenital malformations, deformities and chromosomal abnormalities.

[0051] In embodiments of the present invention, the infectious disease is a prion disease, the neoplasm is a benign or malignant tumor, the endocrine, nutritional and metabolic disease is Wilson's disease, amyloidosis or diabetes mellitus, the mental and behavioral disorder is depression or Rett syndrome, the neurological disease is a nervous system atrophy including central nervous system atrophy, neurodegenerative disease, motor disorder, neuropathy, motor nerve disorder or central nervous system demyelinating disease, the eye and adnexal disease is uveitis, the cardiovascular disease is atrial fibrillation or stroke, the respiratory disease is asthma, the digestive disease is alcoholic liver disease, inflammatory bowel disease, Crohn's disease or ulcerative bowel disease, the skin and subcutaneous tissue disease is psoriasis, the musculoskeletal and connective tissue disease is rheumatoid arthritis, osteoarthritis or systemic lupus erythematosus, and the congenital malformation, deformity and chromosomal abnormality may be autosomal dominant polycystic cystoma.

[0052] In embodiments of the present invention, the systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, may be Huntington's disease, spinal muscular atrophy (SMA), or spinocerebellar ataxia (SCA); the neurodegenerative disease may be Alzheimer's disease or tauopathy; the motor disorder may be Parkinson's disease; the neuropathic disease may be Charcot-Marie-Tooth disease (peripheral Charcot-Marie-Tooth disease, central Charcot-Marie-Tooth disease) or hereditary neuropathy including hereditary spastic paraplegia, diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy, or drug-induced neuropathy; the motor nerve disorder may be amyotrophic lateral sclerosis (ALS); and the central nervous system demyelinating disease may be multiple sclerosis (MS).

[0053] In embodiments of the present invention, diseases related to histone deacetylase 6 (HDAC6) activity or histone deacetylase 6-mediated diseases may include cancer, inflammatory diseases, autoimmune diseases, neurological diseases or neurodegenerative diseases, specifically lung cancer, colon cancer, breast cancer, prostate cancer, liver cancer, brain tumors, ovarian cancer, stomach cancer, skin cancer, pancreatic cancer, glioma, glioblastoma, leukemia, lymphoma, multiple myeloma, solid tumors, Wilson's disease, spinocerebellar ataxia, prion diseases, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, Charcot-Marie-Smith disease. This includes hereditary neuropathy, including Tooth disease (peripheral Charcot-Marie-Tooth disease, central Charcot-Marie-Tooth disease) or hereditary spastic paraplegia, diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy, or drug-induced neuropathy, amyloidosis, Alzheimer's disease, alcoholic liver disease, spinal muscular atrophy, rheumatoid arthritis, or osteoarthritis, and may also include conditions or diseases associated with abnormal function of histone deacetylase 6.

[0054] Chemical formulas I, II, II-1, II-2, and II-3 of the present invention The compounds represented by chemical formula II-4, compounds 1-8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts exhibit high permeability across the blood-brain barrier. Specifically, the compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4 of the present invention, compounds 1-8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts can exhibit a high B / P ratio when administered orally (Table 10).

[0055] The compounds represented by Chemical Formula I, Chemical Formula II, Chemical Formula II-1, Chemical Formula II-2, Chemical Formula II-3, or Chemical Formula II-4 of the present invention, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathy.

[0056] In embodiments of the present invention, the systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative disease, or hereditary neuropathy may be Huntington's disease, spinal muscular atrophy (SMA), spinocerebellar ataxia (SCA), Alzheimer's disease, tauopathy, Charcot-Marie-Tooth disease (including peripheral Charcot-Marie-Tooth disease, central Charcot-Marie-Tooth disease, or hereditary neuropathy including hereditary spastic paraplegia), diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy, drug-induced neuropathy, amyotrophic lateral sclerosis (ALS), or multiple sclerosis (MS).

[0057] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof exhibit remarkably excellent preventive and therapeutic effects against neurodegenerative diseases, including central nervous system system atrophy selected from the group consisting of Huntington's disease, dementia, Alzheimer's disease, amyloidosis, Charcot-Marie-Tooth disease (peripheral Charcot-Marie-Tooth disease, central nervous system Charcot-Marie-Tooth disease), and tauopathy, as well as hereditary neuropathy.

[0058] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1-8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts can increase the relative velocity of mitochondria in axons, which is reduced by treatment of amyloid-beta protein fragments (Aβ), and such pharmacological effects can be sustained over a long period. Therefore, the compounds of the present invention can exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including dementia and systemic atrophy of the central nervous system, neurodegenerative diseases, or hereditary neuropathy (Tables 6, 7, 8, Figures 1 and 2).

[0059] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof can increase the relative velocity of mitochondria in axons, which is reduced in nerve cells overexpressing tau protein. Therefore, the compounds of the present invention can show excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including tauopathy, neurodegenerative diseases, or hereditary neuropathy (Table 9 and Figure 3).

[0060] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1 to 8 listed in Table 1, and Stereoimers of or pharmaceutically acceptable salts of can significantly improve cognitive decline in tauopathy mice (PS19 mice). Therefore, the compounds of the present invention can show excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including tauopathy, neurodegenerative diseases, or hereditary neuropathy (Figures 6, 7, and 8).

[0061] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof can significantly reduce tau hyperphosphorylation observed in the brains of tauopathy mice (PS19 mice). Therefore, the compounds of the present invention can exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including tauopathy, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathy (Figures 9, 10, 11, 12, and 13).

[0062] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof can significantly enhance long-term memory in the hippocampus, which is impaired in tauopathy mice (PS19 mice). Therefore, the compounds of the present invention can show excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including tauopathy, neurodegenerative diseases, or hereditary neuropathy (Figures 14 and 15).

[0063] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof can significantly improve ataxia in Huntington's disease model mice (Yac128 mice) and significantly increase the reduced grip force in Huntington's disease model mice (Yac128 mice). Therefore, the compounds of the present invention can show excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Huntington's disease, neurodegenerative diseases, or hereditary neuropathy (Figures 16, 17, 18, and 19).

[0064] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof are used in the dosal root ganglia of the Charcot-Marie-Tooth disease (CMT) model mouse (MFN2 mutant mouse). The relative velocity of mitochondria in ganglia (DRG) can be significantly increased. Therefore, the compounds of the present invention can show excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Charcot-Marie-Tooth disease, neurodegenerative diseases, or hereditary neuropathy (Table 11 and Figure 20).

[0065] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts can significantly improve the waiting time to drop, which was reduced in Charcot-Marie-Tooth disease (CMT) model mice (CX32 null mice) (confirmed through constant velocity rotorod tests), and can significantly reduce the slip count and beam travel time, which were increased in Charcot-Marie-Tooth disease (CMT) model mice (CX32 null mice) (confirmed through balanced beam tests). Therefore, the present invention can significantly improve the waiting time to drop, which was reduced in Charcot-Marie-Tooth disease (CMT) model mice (CX32 null mice). The compound can demonstrate excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Charcot-Marie-Tooth disease, neurodegenerative diseases, or hereditary neuropathy (Figures 21 and 22).

[0066] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts can significantly improve the waiting time to fall, which was reduced in Charcot-Marie-Tooth disease (CMT) model mice (MFN2 mutant mice) (confirmed through accelerated rotorod testing), and can significantly reduce the slip count, which was increased in Charcot-Marie-Tooth disease (CMT) model mice (MFN2 mutant mice) (confirmed through balanced beam testing). Therefore, the compounds of the present invention can show excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Charcot-Marie-Tooth disease, neurodegenerative diseases, or hereditary neuropathy (Figures 23 and 24).

[0067] In embodiments of the present invention, compounds represented by formula I, formula II, formula II-1, formula II-2, formula II-3, or formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts can significantly improve the waiting time to fall, which was reduced in Charcot-Marie-Tooth disease (CMT) model mice (CMT2A mice) (confirmed through accelerated rotorod testing), and can significantly reduce the slip count, which was increased in Charcot-Marie-Tooth disease (CMT) model mice (CMT2A mice) (confirmed through balanced beam testing). Therefore, the above compounds of the present invention can show excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Charcot-Marie-Tooth disease, neurodegenerative diseases, or hereditary neuropathy (Figures 25 and 26).

[0068] In embodiments of the present invention, compounds represented by formula I, formula II, formula II-1, formula II-2, formula II-3, or formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts can significantly improve the waiting time to fall, which was reduced in Charcot-Marie-Tooth disease (CMT) model mice (CMT1X mice) (confirmed through isokinetic rotorod testing), and can significantly reduce the slip count, which was increased in Charcot-Marie-Tooth disease (CMT) model mice (CMT1X mice) (confirmed through balance beam testing). Therefore, the above compounds of the present invention can show excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Charcot-Marie-Tooth disease, neurodegenerative diseases, or hereditary neuropathy (Figures 27 and 28).

[0069] In embodiments of the present invention, compounds represented by formula I, formula II, formula II-1, formula II-2, formula II-3, or formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts can significantly improve the waiting time to fall, which was reduced in Charcot-Marie-Tooth disease (CMT) model mice (CMT1A mice) (confirmed through isokinetic rotorod testing), and can significantly reduce the slip count, which was increased in Charcot-Marie-Tooth disease (CMT) model mice (CMT1A mice) (confirmed through balance beam testing). Therefore, the above compounds of the present invention can show excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Charcot-Marie-Tooth disease, neurodegenerative diseases, or hereditary neuropathy (Figures 29 and 30).

[0070] In embodiments of the present invention, chemical formulas I, II, II-1, II-2, Compounds represented by chemical formula II-3 or chemical formula II-4, compounds 1-8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts can significantly improve sensory neuron action potential (SNAP) amplitude and sensory neuron conduction velocity (SNCV), which were reduced in Charcot-Marie-Tooth disease (CMT) model mice (MFN2 mutant mice). Therefore, the compounds of the present invention can show excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Charcot-Marie-Tooth disease, neurodegenerative diseases, or hereditary neuropathy (Figure 31).

[0071] In embodiments of the present invention, compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, compounds 1 to 8 listed in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof can significantly increase the axonal size of sciatic nerve fibers, which was reduced in a Charcot-Marie-Tooth disease (CMT) model mouse (CX32 null mouse). Therefore, the compounds of the present invention can exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Charcot-Marie-Tooth disease, neurodegenerative diseases, or hereditary neuropathy (Figure 32).

[0072] Method for producing compounds

[0073] The present invention provides a method for producing an oxadiazole derivative compound represented by chemical formula I, its stereoisomer, or a pharmaceutically acceptable salt thereof.

[0074] In the following reaction equations, those represented by the same symbols as in Chemical Formula I and not specifically explained are the same as those defined in Chemical Formula I, and redundant explanations have been omitted. Furthermore, in the reaction equations, PG represents an amine protecting group, and may be, for example, Boc(tert-Butyloxycarbonyl).

[0075] A preferred method for producing the oxadiazole derivative compound represented by the chemical formula I, its stereoisomer, or a pharmaceutically acceptable salt thereof can be represented by the following reaction formulas 1-1, 1-2, and 1-3, and this includes modified production methods that are obvious to those skilled in the art.

[0076] In embodiments of the present invention, a preferred method for producing an oxadiazole derivative compound represented by chemical formula I, its stereoisomer, or a pharmaceutically acceptable salt thereof can be carried out by the production method shown in reaction formula 1-1 below.

[0077] [ka]

[0078] In the above [Reaction Formula 1-1], X1 to X4, R1, R2 and R3 may be the same as those defined in chemical formula I, PG may be a protecting group, halo may be F, Cl, Br or I, and alkyl may be a C1 to C5 alkyl.

[0079] In embodiments of the present invention, in the above [Reaction Formula 1-1], X1, X3, and X4 are CH, X2 is N, R2 and R3 may each be independently F or Cl, R1 may be CF2H, PG may be tert-butylcarboxylic acid, and alkyl may be methyl, ethyl, or butyl.

[0080] In the above [reaction formula 1-1], the compounds represented by 1-1-1, 1-1-2, 1-1-3, 1-1-4, 1-1-5, 1-1-6, 1-1-7, 1-1-8, 1-1-9, or 1-1-10 may each be independently in the form of a salt, and the salt may be a hydrochloride salt or a trifluoroacetate salt.

[0081] The aforementioned [Reaction Formula 1-1] involves using a method for synthesizing 1,3,4-oxadiazole derivative compounds to react a compound of chemical formula 1-1-1 containing an isocyanate with a compound of chemical formula 1-1-2 to which a protecting group has been introduced, thereby producing a compound of chemical formula 1-1-3 containing a urea structure.

[0082] Subsequently, the compound of chemical formula 1-1-5 is produced by a substitution reaction with the compound of chemical formula 1-1-4, and then the protecting group is removed to produce the compound of chemical formula 1-1-6.

[0083] The compound of chemical formula 1-1-6 is first reacted with the compound of chemical formula 1-1-7 in a reductive amination reaction to produce the compound of chemical formula 1-1-8, which is then reacted with hydrazine to produce the hydrazide compound of chemical formula 1-1-9. Subsequently, the compound of chemical formula 1-1-10 can be produced using difluoroacetic anhydride and imidazole.

[0084] In embodiments of the present invention, the compound of chemical formula 1-1-10 in the reaction formula [reaction formula 1-1] may be compound 3.

[0085] In embodiments of the present invention, a preferred method for producing the oxadiazole derivative compound represented by chemical formula I, its isomers, or pharmaceutically acceptable salts thereof can be carried out by the following production method of reaction 1-2.

[0086] [ka]

[0087] In the above [reaction formula 1-2], X1 to X4, R1, R2 and R3 may be the same as those defined in chemical formula I, PG may be a protecting group, halo may be F, Cl, Br or I, and alkyl may be a C1 to C5 alkyl.

[0088] In embodiments of the present invention, in the above [reaction formula 1-2], X1, X3, and X4 are CH, X2 is N or -CRx (Rx is F or Cl, Br or I), R2 and R3 may each be independently F or Cl, R1 may be CF2H, PG may be tert-butylcarboxylic acid, and alkyl may be methyl, ethyl or butyl.

[0089] In the above [reaction formula 1-2], the compounds represented by 1-1-1, 1-2-1, 1-2-2, 1-1-4, 1-1-8, 1-1-9, or 1-1-10 may each independently be in the form of a salt, and the salt may be a hydrochloride salt or a trifluoroacetate salt.

[0090] The aforementioned [reaction formula 1-2] involves using a method for synthesizing 1,3,4-oxadiazole compounds having an oxetane structure to react a compound of chemical formula 1-1-1 containing an isocyanate with a compound of chemical formula 1-2-1 in which an oxetane has been introduced, thereby producing a compound of chemical formula 1-2-2 containing a urea structure.

[0091] The compound of chemical formula 1-2-2 is subjected to a substitution reaction with the compound of chemical formula 1-1-4 to produce the compound of chemical formula 1-1-8, which is then reacted with hydrazine to produce the hydrazide compound of chemical formula 1-1-9. Subsequently, the desired compound of chemical formula 1-1-10 is produced using difluoroacetic anhydride and imidazole.

[0092] In embodiments of the present invention, the compound of chemical formula 1-1-10 in [reaction formula 1-2] may be compound 4, compound 5, compound 6, compound 7, or compound 8, etc.

[0093] In embodiments of the present invention, a preferred method for producing the oxadiazole derivative compound represented by chemical formula I, its isomers, or pharmaceutically acceptable salts thereof can be carried out by the following production method shown in reaction formulas 1-3.

[0094] [ka]

[0095] In the above [Reaction Equation 1-3], X1 to X4, R1, R2 and R3 are defined by chemical formula I. It may be the same as defined, PG may be a protecting group, halo may be F, Cl, Br or I, and alkyl may be a C1-C5 alkyl group.

[0096] In embodiments of the present invention, in the above [reaction formula 1-3], X1, X3, and X4 are CH, X2 is CH or -CRx (Rx is F or Cl, Br or I), R2 and R3 may each be independently F or Cl, R1 may be CF2H, PG may be tert-butylcarboxylic acid, and alkyl may be methyl, ethyl or butyl.

[0097] In the above [reaction formula 1-3], the compounds represented by 1-3-1, 1-1-2, 1-1-4, 1-3-2, 1-1-5, 1-1-6, 1-1-7, 1-1-8, 1-1-9, or 1-1-10 may each be independently in the form of a salt, and the salt may be a hydrochloride salt or a trifluoroacetate salt.

[0098] The aforementioned [reaction formula 1-3] is a synthesis method for oxadiazole compounds having an oxetane structure. A compound of chemical formula 1-3-1 containing an amine group is substituted with a compound of chemical formula 1-1-4 to produce a compound of chemical formula 1-3-2. This compound is then reacted with the compound of chemical formula 1-1-2, which has a protecting group introduced, to produce a compound of chemical formula 1-1-5 containing a urea structure. Subsequently, the protecting group of chemical formula 1-1-5 is removed to produce a compound of chemical formula 1-1-6.

[0099] The compound of chemical formula 1-1-6 is first reacted with the compound of chemical formula 1-1-7 in a reductive amination reaction to produce the compound of chemical formula 1-1-8, which is then reacted with hydrazine to produce the hydrazide compound of chemical formula 1-1-9. Subsequently, the desired compound of chemical formula 1-1-10 is produced using difluoroacetic anhydride and imidazole.

[0100] In embodiments of the present invention, the compound of chemical formula 1-1-10 in [reaction formula 1-3] may be compound 1 or compound 2, etc.

[0101] Pharmaceutical composition, therapeutic method using the same, and its use The present invention provides a pharmaceutical composition comprising a compound represented by the chemical formula I, its stereoisomer, or a pharmaceutically acceptable salt thereof.

[0102] According to embodiments of the present invention, the compound represented by chemical formula I contained in the pharmaceutical composition may be the compound represented by chemical formula II.

[0103] According to embodiments of the present invention, the compound represented by chemical formula I contained in the pharmaceutical composition may be a compound represented by chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4.

[0104] The present invention provides a pharmaceutical composition comprising at least one compound from compounds 1 to 8 listed in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0105] The present invention provides a pharmaceutical composition comprising at least one compound from compounds 1 and 5 listed in Table 1, its stereoisomer, or a pharmaceutically acceptable salt thereof.

[0106] The present invention provides a compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, its stereoisomer, or a pharmaceutically acceptable salt thereof as an active ingredient. The present invention provides a pharmaceutical composition for the prevention or treatment of histone deacetylase-mediated diseases, which includes the following:

[0107] The present invention provides a pharmaceutical composition for the prevention or treatment of histone deacetylase 6-mediated diseases, comprising as an active ingredient a compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0108] The pharmaceutical compositions of the present invention can exhibit the same type of pharmacological effects as those exhibited by compounds represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, its stereoisomer, or a pharmaceutically acceptable salt thereof.

[0109] In the aforementioned pharmaceutical composition, the histone deacetylase 6-mediated disease may be substantially the same as the histone deacetylase 6-mediated disease of the aforementioned compound.

[0110] In embodiments of the present invention, diseases or mediated by histone deacetylase 6 (HDAC6) activity may include infectious diseases, neoplasms, endocrine, nutritional and metabolic disorders, mental and behavioral disorders, neurological disorders, ocular and adnexal disorders, cardiovascular diseases, respiratory diseases, digestive diseases, skin and subcutaneous tissue disorders, musculoskeletal and connective tissue disorders, or congenital malformations, deformities and chromosomal abnormalities.

[0111] In embodiments of the present invention, the infectious disease is a prion disease, the neoplasm is a benign or malignant tumor, the endocrine, nutritional and metabolic disease is Wilson's disease, amyloidosis or diabetes mellitus, the mental and behavioral disorder is depression or Rett syndrome, the neurological disease is a nervous system atrophy including central nervous system atrophy, neurodegenerative disease, motor disorder, neuropathy, motor nerve disorder or central nervous system demyelinating disease, the ocular and adnexal disease is uveitis, the cardiovascular disease is atrial fibrillation or stroke, the respiratory disease is asthma, the digestive disease is alcoholic liver disease, inflammatory bowel disease, Crohn's disease or ulcerative bowel disease, the skin and subcutaneous tissue disease is psoriasis, the musculoskeletal and connective tissue disease is rheumatoid arthritis, osteoarthritis or systemic lupus erythematosus, and the congenital malformation, deformity and chromosomal abnormality may be autosomal dominant polycystic cystoma.

[0112] In embodiments of the present invention, the systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, may be Huntington's disease, spinal muscular atrophy (SMA), or spinocerebellar ataxia (SCA); the neurodegenerative disease may be Alzheimer's disease or tauopathy; the motor disorder may be Parkinson's disease; the neuropathic disease may be Charcot-Marie-Tooth disease (peripheral Charcot-Marie-Tooth disease, central Charcot-Marie-Tooth disease), or hereditary neuropathy including hereditary spastic paraplegia, diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy, or drug-induced neuropathy; the motor nerve disorder may be amyotrophic lateral sclerosis (ALS); and the demyelinating disease of the central nervous system may be multiple sclerosis (MS).

[0113] In embodiments of the present invention, diseases or mediated by histone deacetylase 6 (HDAC6) activity may include cancer, inflammatory diseases, autoimmune diseases, neurological diseases, or neurodegenerative diseases, specifically lung cancer, colon cancer, breast cancer, prostate cancer, liver cancer, brain tumors, ovarian cancer, stomach cancer, skin cancer, pancreatic cancer, glioma, glioblastoma, leukemia, lymphoma, multiple myeloma, solid tumors, and Wilson's disease. This includes hereditary neuropathy, including spinocerebellar ataxia, prion diseases, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, Charcot-Marie-Tooth disease (peripheral Charcot-Marie-Tooth disease, central Charcot-Marie-Tooth disease), or hereditary spastic paraplegia, diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy, or drug-induced neuropathy, amyloidosis, Alzheimer's disease, alcoholic liver disease, spinal muscular atrophy, rheumatoid arthritis, or osteoarthritis, and may also include conditions or diseases associated with abnormal function of histone deacetylase 6.

[0114] The present invention provides a pharmaceutical composition for the prevention or treatment of systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathy, comprising as an active ingredient a compound represented by the aforementioned chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0115] In the aforementioned pharmaceutical composition, systemic atrophy of the nervous system, neurodegenerative diseases, and hereditary neuropathy may be substantially the same as the degenerative brain diseases described above in the description of the compounds.

[0116] In embodiments of the present invention, the degenerative brain disease may be Huntington's disease, spinal muscular atrophy (SMA), spinocerebellar ataxia (SCA), Alzheimer's disease, tauopathy, Charcot-Marie-Tooth disease (peripheral Charcot-Marie-Tooth disease, central Charcot-Marie-Tooth disease), or hereditary neuropathy including hereditary spastic paraplegia, diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy, drug-induced neuropathy, amyotrophic lateral sclerosis (ALS), or multiple sclerosis (MS). Specifically, the degenerative brain disease may be Huntington's disease, dementia, Alzheimer's disease, amyloidosis, or tauopathy.

[0117] In the aforementioned pharmaceutical composition, the stereoisomers and pharmaceutically acceptable salts are as described in the section on stereoisomers and pharmaceutically acceptable salts of the compound described above.

[0118] The pharmaceutical composition of the present invention may further contain one or more pharmaceutically acceptable additives, in addition to the compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0119] In embodiments of the present invention, pharmaceutically acceptable additives may include physiological saline, sterile water, Ringer's solution, buffered physiological saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of these components. Other common additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed. Diluents, dispersants, surfactants, binders, and lubricants may also be added, and the formulations can be prepared as injectable dosage forms such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, or tablets. Therefore, the compositions of the present invention may be patches, liquids, pills, capsules, granules, tablets, suppositories, etc. These formulations can be produced by conventional methods used in formulation in the art or by methods disclosed in Remington's Pharmaceutical Science (latest edition), Mack Publishing Company, Easton PA, and can be formulated into a variety of formulations depending on the disease or component.

[0120] In embodiments of the present invention, pharmaceutically acceptable additives that the pharmaceutical composition may contain are those commonly used in the art, specifically lactose, dextrose, sucrose, sorbitol, mannitol, glycine, starch, tragacanth gum, and red sugar. Examples of ingredients include, but are not limited to, shea gum, calcium phosphate, calcium chloride, sodium chloride, alginic acid, sodium alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, polyethylene glycol, cellulose, water, ethanol, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoic acid, talc, magnesium stearate, magnesium aluminum silicate, silica, orange essence, strawberry essence, vanilla essence, or mineral oil.

[0121] The pharmaceutical composition of the present invention may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) by the method of the present invention, and the dosage may vary depending on the patient's weight, age, sex, health condition, diet, time of administration, method of administration, route of administration, excretion rate, type of disease, severity of disease, duration of treatment, including drugs used in combination or concurrently, and other factors well known in the medical field, and may be determined by a person skilled in the art taking the above factors into consideration.

[0122] The daily dose of a pharmaceutical composition comprising a compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4 of the present invention, at least one compound from compounds 1 to 8 listed in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is approximately 1 to 1000 mg / kg, preferably 5 to 100 mg / kg, and may be administered once or in several divided doses per day.

[0123] The pharmaceutical composition of the present invention may further contain, in addition to the compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, its stereoisomer or a pharmaceutically acceptable salt thereof, one or more active ingredients exhibiting the same or similar pharmacological effects.

[0124] The present invention provides a method for preventing or treating diseases related to HDAC6 activity, or neurodegenerative diseases including central nervous system system atrophy, neurodegenerative brain diseases including degenerative brain diseases, or hereditary neuropathy, comprising the step of administering to a subject a compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same.

[0125] The present invention provides a method for preventing or treating diseases related to the activity of histone deacetylase 6 or degenerative brain diseases, which includes not only treating the disease itself before the onset of symptoms, but also inhibiting or avoiding its symptoms by administering a compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, its stereoisomer, or a pharmaceutically acceptable salt thereof. In the management of the disease, the prophylactic or therapeutic dose of a particular active ingredient varies depending on the nature and severity of the disease or condition, and the route through which the active ingredient is administered. The dose and frequency of administration will vary depending on the age, weight, and response of the individual patient. Appropriate dosage and administration can be easily selected by a person with ordinary skill in the art, taking these factors into consideration.

[0126] In the method of the present invention, the disease or degenerative brain disease related to the activity of histone deacetylase 6 is the same as that described above.

[0127] In this invention, "administration" means introducing a given substance into a target in an appropriate manner.

[0128] In the present invention, "subject" means any animal, including mice, rats, and livestock, including humans who have or are likely to develop a disease or degenerative brain disease related to HDAC6 activity, and may specifically be mammals, including humans.

[0129] The present invention provides a method for preventing or treating diseases related to the activity of HDAC6, or systemic atrophy of the nervous system including central nervous system system atrophy, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathy, by administering a therapeutically effective dose of a compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0130] In the present invention, "therapeutic effective dose" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment and without causing side effects, and refers to an amount of a compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, its stereoisomer, or a pharmaceutically acceptable salt thereof, which is effective in preventing or treating diseases related to the activity of histone deacetylase 6 or systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathy.

[0131] Furthermore, the present invention provides a method for preventing or treating diseases related to the activity of histone deacetylase 6, or systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathy, which may further include administering an additional active agent in a therapeutically effective amount that helps treat the disease in combination with the compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, its stereoisomer, or a pharmaceutically acceptable salt thereof, and the additional active agent may exhibit a synergistic, additive, or auxiliary effect together with the compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, its stereoisomer, or a pharmaceutically acceptable salt thereof.

[0132] The present invention provides uses for a compound represented by chemical formula I, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, for the prevention or treatment of diseases related to HDAC6 activity or systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathy.

[0133] The present invention provides for the use of compounds represented by Chemical Formula I, Chemical Formula II, Chemical Formula II-1, Chemical Formula II-2, Chemical Formula II-3, or Chemical Formula II-4, at least one compound from Compounds 1 to 8 listed in Table 1, stereoisomers thereof or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing thereof, for the manufacture of agents for the prevention or treatment of diseases related to HDAC6 activity or systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathy.

[0134] In the aforementioned uses of the present invention, diseases related to the activity of histone deacetylase 6, or systemic atrophy of the nervous system including systemic atrophy of the central nervous system, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathy are the same as those described above.

[0135] For the manufacture of pharmaceuticals, a compound represented by chemical formula I of the present invention, chemical formula II, chemical formula II-1, chemical formula II-2, chemical formula II-3, or chemical formula II-4, at least one compound from compounds 1 to 8 listed in Table 1, its stereoisomer, or a pharmaceutically acceptable salt thereof may be mixed with a pharmaceutically acceptable adjuvant, diluent, carrier, etc., and may be manufactured as a complex formulation together with other active preparations to exhibit a synergistic effect.

[0136] Each item of the present invention, namely oxadiazole derivative compounds, methods for producing the same, pharmaceutical compositions containing the same, therapeutic methods using the same, and their uses, is applicable equally to each other, as long as they do not contradict each other. [Effects of the Invention]

[0137] The novel oxadiazole derivative compounds of the present invention, their stereoisomers, or pharmaceutically acceptable salts thereof possess selective HDAC6 inhibitory activity and exhibit excellent preventive or therapeutic effects against diseases associated with HDAC6 activity or systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathy. [Brief explanation of the drawing]

[0138] [Figure 1] The results of evaluating the effect of the compound of the present invention on the migration rate of mitochondria within axons, which was reduced by Aβ treatment in mouse hippocampal neurons, are shown. [Figure 2] The results of evaluating the effect of the compound of the present invention on the migration rate of mitochondria within axons, which was reduced by Aβ treatment in mouse hippocampal neurons, are shown. [Figure 3] This paper presents the results of evaluating the effect of the compound of the present invention on the reduced mitochondrial migration rate within axons in primary mouse cultured cells overexpressing human tau protein with the P301L mutation. [Figure 4] The results of evaluating the effect of the compound of the present invention on tubulin acetylation in SH-SY5Y cells, a type of human neuroblastoma, are shown. [Figure 5] The results of evaluating the effect of the compound of the present invention on tubulin acetylation in SH-SY5Y cells, a type of human neuroblastoma, are shown. [Figure 6] The results of evaluating the effect of the compound of the present invention on cognitive decline due to tauopathy in PS19 mice are shown. [Figure 7] The results of evaluating the effect of the compound of the present invention on cognitive decline due to tauopathy in PS19 mice are shown. [Figure 8] The results of evaluating the effect of the compound of the present invention on cognitive decline due to tauopathy in PS19 mice are shown. [Figure 9] The results of evaluating the effect of the compound of the present invention on tau hyperphosphorylation in the brains of PS19 mice are shown. [Figure 10] The results of evaluating the effect of the compound of the present invention on tau hyperphosphorylation in the brains of PS19 mice are shown. [Figure 11] The results of evaluating the effect of the compound of the present invention on tau hyperphosphorylation in the brains of PS19 mice are shown. [Figure 12]The results of evaluating the effect of the compound of the present invention on tau hyperphosphorylation in the brains of PS19 mice are shown. [Figure 13] The results of evaluating the effect of the compound of the present invention on tau hyperphosphorylation in the brains of PS19 mice are shown. [Figure 14] The results of evaluating the effect of the compound of the present invention on enhancing long-term memory in the hippocampus, which has deteriorated in PS19 mice, are shown. [Figure 15] The results of evaluating the effect of the compound of the present invention on enhancing long-term memory in the hippocampus, which has deteriorated in PS19 mice, are shown. [Figure 16] The results of evaluating the effect of the compound of the present invention on reduced motor function in Yac128 mice are shown. [Figure 17] The results of evaluating the effect of the compound of the present invention on reduced motor function in Yac128 mice are shown. [Figure 18] The results of evaluating the effect of the compound of the present invention on reduced motor function in Yac128 mice are shown. [Figure 19] The results of evaluating the effect of the compound of the present invention on reduced motor function in Yac128 mice are shown. [Figure 20] The results of evaluating the effect of the compound of the present invention on the reduced mitochondrial migration rate in axons of Charcot-Marie-Tooth disease animal model (MFN2 mutant mice) are shown. [Figure 21] The results of evaluating the effects of the compound of the present invention on reduced motor and sensory functions (rotarod test, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CX32 null mouse) are shown. [Figure 22] The results of evaluating the effects of the compound of the present invention on reduced motor and sensory functions (rotarod test, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CX32 null mouse) are shown. [Figure 23]The results of evaluating the effects of the compound of the present invention on reduced motor and sensory functions (rotarod test, balance beam test) in an animal model of Charcot-Marie-Tooth disease (MFN2 mutant mouse) are shown. [Figure 24] The results of evaluating the effects of the compound of the present invention on reduced motor and sensory functions (rotarod test, balance beam test) in an animal model of Charcot-Marie-Tooth disease (MFN2 mutant mouse) are shown. [Figure 25] The results of evaluating the effects of the compound of the present invention on reduced motor and sensory functions (rotarod test, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CMT2A mouse) are shown. [Figure 26] The results of evaluating the effects of the compound of the present invention on reduced motor and sensory functions (rotarod test, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CMT2A mouse) are shown. [Figure 27] The results of evaluating the effects of the compound of the present invention on reduced motor and sensory functions (rotarod test, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CMT1X mouse) are shown. [Figure 28] The results of evaluating the effects of the compound of the present invention on reduced motor and sensory functions (rotarod test, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CMT1X mouse) are shown. [Figure 29] The results of evaluating the effects of the compound of the present invention on reduced motor and sensory functions (rotarod test, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CMT1A mouse) are shown. [Figure 30] The results of evaluating the effects of the compound of the present invention on reduced motor and sensory functions (rotarod test, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CMT1A mouse) are shown. [Figure 31]The results of evaluating the effect of the compound of the present invention on the reduced nerve conduction velocity in an animal model of Charcot-Marie-Tooth disease (MFN2 mutant mice) are shown. [Figure 32] The results of evaluating the effect of the compound of the present invention on the reduced axonal size of sciatic nerve fibers in an animal model of Charcot-Marie-Tooth disease (CX32 null mice) are shown. [Modes for carrying out the invention]

[0139] The present invention will be described in more detail below with reference to embodiments. These embodiments are for illustrative purposes only, and it will be obvious to those who are ordinarily skilled in the art that the scope of the present invention is not limited by these embodiments. Ro. [Examples]

[0140] Manufacturing of compounds The specific method for producing the compound represented by chemical formula I is as follows:

[0141] Example 1: Synthesis of Compound 1, N-(3-chloro-4-fluorophenyl)-N-(4-(5-(difluoromethyl)-1,3,4-oxadiazole-2-yl)benzyl)-4-(oxetan-3-yl)piperazine-1-carboxamide

[0142] [Step 1] Synthesis of methyl 4-(((3-chloro-4-fluorophenyl)amino)methyl)benzoic acid [ka]

[0143] A solution of 3-chloro-4-fluoroaniline (2.911 g, 20.000 mmol) and sodium hydride (60.00%, 0.880 g, 22.000 mmol) dissolved in N,N-dimethylformamide (80 mL) was stirred at room temperature for 30 minutes. Methyl 4-(bromomethyl)benzoic acid (4.581 g, 20.000 mmol) was added, and the mixture was stirred further at the same temperature for 18 hours. Water was added to the reaction mixture, and it was extracted with ethyl acetate. The organic layer was washed with saturated sodium chloride aqueous solution, water was removed with anhydrous magnesium sulfate, and the mixture was filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography (SiO2, 40 g cartridge, ethyl acetate / hexane = 0%~10%), concentrated, and the title compound (3.711 g, 63.2%) was obtained as a brown oily substance.

[0144] [Step 2] Synthesis of tert-butyl 4-((3-chloro-4-fluorophenyl)(4-(methoxycarbonyl)benzyl)carbamoyl)piperazine-1-carboxylate [ka]

[0145] The solution of methyl 4-(((3-chloro-4-fluorophenyl)amino)methyl)benzoic acid (3.711 g, 12.635 mmol), N,N-diisopropylethylamine (4.401 mL, 25.270 mmol), and triphosgene (1.875 g, 6.317 mmol) prepared in Step 1, dissolved in dichloromethane (50 mL), was stirred at room temperature for 10 minutes. Then, tert-butylpiperazine-1-carboxylate (2.353 g, 12.635 mmol) was added, and the mixture was stirred further at the same temperature for 18 hours. After removing the solvent from the reaction mixture under reduced pressure, the title compound was used without further purification (6.300 g, 98.5%, brown oily substance).

[0146] [Step 3] Synthesis of methyl 4-((N-(3-chloro-4-fluorophenyl)piperazine-1-carboxamide)methyl)benzoate [ka]

[0147] The tert-butyl 4-((3-chloro-4-fluorophenyl)(4-(methoxycarbonyl)benzyl)carbamoyl)piperazine-1-carboxylate (6.300 g, 12.451 mmol) prepared in Step 2 and hydrogen chloride (4.00 M solution in 1,4-dioxane, 12.451 mL, 49.805 mmol) were dissolved in dichloromethane (50 mL) at room temperature and the solution was stirred at the same temperature for 3 hours. The precipitated solid was filtered, washed with dichloromethane, and dried to obtain the title compound (3.647 g, 66.2%) as a white solid.

[0148] [Step 4] Synthesis of methyl 4-((N-(3-chloro-4-fluorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide)methyl)benzoic acid [ka]

[0149] The solution prepared in Step 3 by dissolving methyl 4-((N-(3-chloro-4-fluorophenyl)piperazine-1-carboxamide)methyl)benzoate (0.885 g, 2.000 mmol), oxetane-3-one (0.234 mL, 4.000 mmol), and sodium triacetoxyborohydride (0.848 g, 4.000 mmol) in dichloromethane (10 mL) at room temperature was stirred for 18 hours. Water was added to the reaction mixture, and after extraction with dichloromethane, the mixture was filtered through a plastic filter to remove the solid residue and aqueous layer, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (SiO2, 12 g cartridge; methanol / dichloromethane = 0%~5%) and concentrated to obtain the title compound (0.626 g, 67.7%) as a brown oily substance.

[0150] [Step 5] Synthesis of N-(3-chloro-4-fluorophenyl)-N-(4-(hydrazinecarbonyl)benzyl)-4-(oxetan-3-yl)piperazine-1-carboxamide [ka]

[0151] The methyl 4-((N-(3-chloro-4-fluorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide)methyl)methyl benzoate (0.626 g, 1.355 mmol) prepared in Step 4 and hydrazine monohydrate (1.317 mL, 27.104 mmol) were dissolved in ethanol (6 mL) at room temperature. The solution was stirred at 75 °C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. After removing the solvent from the reaction mixture under reduced pressure, the concentrate was purified by column chromatography (SiO2, 12 g cartridge; methanol / dichloromethane = 0%~10%) and concentrated to obtain the title compound (0.435 g, 69.5%) as a white solid.

[0152] [Step 6] Synthesis of Compound 1 [ka]

[0153] The solution prepared in Step 5 by dissolving N-(3-chloro-4-fluorophenyl)-N-(4-(hydrazinecarbonyl)benzyl)-4-(oxetan-3-yl)piperazine-1-carboxamide (0.100 g, 0.216 mmol), triethylamine (0.091 mL, 0.649 mmol), and 2,2-difluoroacetic anhydride (0.081 mL, 0.649 mmol) in dichloromethane (2 mL) at room temperature was stirred at 40°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Water was added to the reaction mixture, extracted with dichloromethane, filtered through a plastic filter to remove the solid residue and aqueous layer, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (SiO2, 4g cartridge; ethyl acetate / hexane = 0%~60%) and concentrated to obtain title compound 1 (0.099g, 87.9%) as a white solid.

[0154] 1H NMR (400 MHz, CDCl3) δ 8.07 - 8.04 (m, 2H), 7.46 (d, 2H, J = 8.5 Hz), 7.15 (dd, 1H, J = 6.3, 2.7 Hz), 7.10 (t, 1H, J = 8.6 Hz), 7.05 - 6.80 (m, 2H), 4.90 (s, 2H), 4.77 - 4.74 (m, LRMS (ES) m / z 522.4 (M + + 1).

[0155] Example 2: Synthesis of Compound 2, N-(3-chloro-4-fluorophenyl)-N-(4-(5-(difluoromethyl)-1,3,4-oxadiazole-2-yl)-2-fluorobenzyl)-4-(oxetan-3-yl)piperazine-1-carboxamide Compound 2 was prepared according to the same reaction described in steps 1 to 6 of Example 1, except that methyl 4-(bromomethyl)-3-fluorobenzoic acid was used instead of methyl 4-(bromomethyl)benzoic acid in step 1 of Example 1.

[0156] 1 H NMR (400 MHz, CDCl3) δ 7.89 (dd, 1H, J = 8.0, 1.6 Hz), 7.77 (dd, 1H, J = 10.1, 1.6 Hz), 7.68 (t, 1H, J = 7.6 Hz), 7.19 (dd, 1H, J = 6.3, 2.7 Hz), 7.12 (t, 1H, J = 8.6 Hz), 7.06 - 6.80 (m, 2H), 4.92 (s, 2H), 4.75 - 4.72 (m, LRMS(ES) m / z 540.4 (M + + 1).

[0157] Example 3: Synthesis of Compound 3, N-(3-chloro-4-fluorophenyl)-N-((5-(5-(difluoromethyl)-1,3,4-oxadiazole-2-yl)pyridine-2-yl)methyl)-4-(oxetan-3-yl)piperazine-1-carboxamide [Step 1] Synthesis of tert-butyl 4-((3-chloro-4-fluorophenyl)((5-(methoxycarbonyl)pyridine-2-yl)methyl)carbamoyl)piperazine-1-carboxylate [ka]

[0158] 1.500 g, 4.192 mmol of tert-butyl 4-((3-chloro-4-fluorophenyl)carbamoyl)piperazine-1-carboxylate and sodium hydride (60.00%, 0.184 g, 4.611 mmol) were dissolved in N,N-dimethylformamide (30 mL) at 0°C. Methyl 6-(bromomethyl)nicotinate (1.013 g, 4.402 mmol) was added to this solution, and the mixture was stirred at the same temperature for 1 hour. A saturated sodium bicarbonate aqueous solution was poured over the reaction mixture, and it was extracted with ethyl acetate. The organic layer was washed with a saturated sodium chloride aqueous solution, water was removed with anhydrous magnesium sulfate, and the mixture was filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography (SiO2, 24 g cartridge; ethyl acetate / hexane = 20%~60%), concentrated, and the title compound 1 (2.000 g, 94.1%) was obtained as a yellow solid.

[0159] [Step 2] Synthesis of methyl 6-((N-(3-chloro-4-fluorophenyl)piperazine-1-carboxamide)methyl)nicotinate hydrochloride [ka]

[0160] The tert-butyl 4-((3-chloro-4-fluorophenyl)((5-(methoxycarbonyl)pyridine-2-yl)methyl)carbamoyl)piperazine-1-carboxylate (2.000 g, 3.945 mmol) prepared in Step 1 and hydrochloric acid (4.00 M solution in 1,4-dioxane, 4.931 mL, 19.725 mmol) were dissolved in dichloromethane (50 mL) at room temperature and the solution was stirred at the same temperature for 18 hours. Saturated sodium bicarbonate aqueous solution was poured over the reaction mixture and extracted with dichloromethane. The organic layer was washed with saturated sodium chloride aqueous solution, water was removed with anhydrous sodium sulfate, and the mixture was filtered and concentrated under reduced pressure. Ethyl acetate was added to the concentrate and stirred. The precipitated solid was filtered, washed with ethyl acetate, and dried to obtain the title compound (1.120 g, 64.0%) as a yellow solid.

[0161] [Step 3] Synthesis of methyl 6-((N-(3-chloro-4-fluorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide)methyl)nicotinate [ka]

[0162] In Step 2, methyl 6-((N-(3-chloro-4-fluorophenyl)piperazine-1-carboxamide)methyl)nicotinate hydrochloride (1.120 g, 2.527 mmol) and N,N-diisopropylethylamine (0.440 mL, 2.527 mmol) were prepared by dissolving them in dichloromethane (20 mL) at room temperature. Oxetane-3-one (0.194 mL, 3.032 mmol) was added to the solution and the mixture was stirred at the same temperature. Sodium triacetoxyborohydride (0.803 g, 3.790 mmol) was added to the reaction mixture and the mixture was stirred for a further 18 hours at the same temperature. A saturated aqueous sodium bicarbonate solution was poured over the reaction mixture, extracted with dichloromethane, filtered through a plastic filter to remove the solid residue and aqueous layer, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (SiO2, 24 g cartridge, ethyl acetate = 100%), concentrated, and then purified again by chromatography (SiO2, 24 g cartridge; methanol / dichloromethane = 0% to 10%), concentrated, to obtain the title compound (0.467 g, 39.9%) as a yellow oil.

[0163] [Step 4] Synthesis of N-(3-chloro-4-fluorophenyl)-N-((5-(hydrazinecarbonyl)pyridine-2-yl)methyl)-4-(oxetan-3-yl)piperazine-1-carboxamide [ka]

[0164] The methyl 6-((N-(3-chloro-4-fluorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide)methyl)nicotinate (0.467 g, 1.009 mmol) prepared in Step 3 and hydrazine monohydrate (0.981 mL, 20.177 mmol) were dissolved in ethanol (4 mL) at room temperature. The solution was stirred at 110 °C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. After removing the solvent from the reaction mixture under reduced pressure, the title compound was used without further purification (0.460 g, 98.5%, yellow solid).

[0165] [Step 5] Synthesis of Compound 3 [ka]

[0166] In Step 4, N-(3-chloro-4-fluorophenyl)-N-((5-(hydrazinecarbonyl)pyridine-2-yl)methyl)-4-(oxetan-3-yl)piperazine-1-carboxamide (0.470 g, 1.015 mmol) and imidazole (0.207 g, 3.046 mmol) were prepared by dissolving them in dichloromethane (10 mL) at room temperature. To this solution, 2,2-difluoroacetic anhydride (0.379 mL, 3.046 mmol) was added, and the mixture was heated under reflux for 18 hours. After that, the temperature was lowered to room temperature to terminate the reaction. A saturated aqueous sodium bicarbonate solution was poured over the reaction mixture, and after extraction with dichloromethane, the mixture was filtered through a plastic filter to remove the solid residue and aqueous layer, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (SiO2, 12g cartridge; methanol / dichloromethane = 0%~2.5%) and concentrated to obtain compound 3 (0.167g, 31.5%) as a yellow solid.

[0167] 1 H NMR (400 MHz, CDCl3) δ 9.25 (dd, J = 2.2, 0.7 Hz, 1H), 8.34 (dd, J = 20.0, 22.2 Hz, 1H ), 7.60 (d, J = 8.2 Hz, 1H), 7.29 - 7.27 (m, 2H), 7.13 - 6.83 (m, 3H), 5.06 (s, 2H), 4.65 (t, J = 6.6 Hz, 2H), 4.56 (t, J = 6.0 Hz, 2H), 3.46 - 3.43 (m, 1H), 3.34 - 3.33 (m, 4H), 2.19 - 2.18 (m, 4H); LRMS (ES) m / z 523.3 (M + + 1 ).

[0168] Example 4: Synthesis of Compound 4, N-(3,4-dichlorophenyl)-N-(4-(5-(difluoromethyl)-1,3,4-oxadiazole-2-yl)-2-fluorobenzyl)-4-(oxetan-3-yl)piperazine-1-carboxamide [Step 1] Synthesis of tert-butyl 4-(oxetan-3-yl)piperazine-1-carboxylate [ka]

[0169] 5.000 g, 26.844 mmol of tert-butylpiperazine-1-carboxylate and 2.902 g, 40.266 mmol of oxetane-3-one were dissolved in 200 mL of dichloromethane at room temperature. Sodium triacetoxyborohydride (11.379 g, 53.688 mmol) was added to this solution, and the mixture was stirred at the same temperature for 18 hours. A saturated aqueous sodium bicarbonate solution was poured over the reaction mixture, and it was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, water was removed with anhydrous magnesium sulfate, and the mixture was filtered and concentrated under reduced pressure. Diethyl ether was added to the concentrate and stirred. The precipitated solid was filtered, washed with diethyl ether, and dried to obtain the title compound (6.230 g, 95.8%) as a white solid.

[0170] [Step 2] Synthesis of 1-(oxetan-3-yl)piperazine trifluoroacetate [ka]

[0171] A solution of tert-butyl 4-(oxetan-3-yl)piperazine-1-carboxylate (6.230 g, 25.710 mmol) prepared in Step 1 and trifluoroacetic acid (5.906 mL, 77.129 mmol) dissolved in dichloromethane (50 mL) at room temperature was stirred at the same temperature for 7 hours. After removing the solvent from the reaction mixture under reduced pressure, ethyl acetate was added to the concentrate and stirred. The precipitated solid was filtered, washed with ethyl acetate, and dried to obtain the title compound (4.720 g, 76.7%) as a white solid.

[0172] [Step 3] Synthesis of N-(3,4-dichlorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide [ka]

[0173] The solution, prepared by dissolving 3,4-dichlorophenyl isocyanate (0.300 g, 1.596 mmol) and 1-(oxetan-3-yl)piperazine 2,2,2-trifluoroacetate (0.409 g, 1.596 mmol) prepared in Step 2 in diethyl ether (10 mL) at room temperature, was stirred at the same temperature for 2 hours. The precipitated solid was filtered, washed with diethyl ether, and dried to obtain the title compound (0.497 g, 94.3%) as a white solid.

[0174] [Step 4] Synthesis of N-(3,4-dichlorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide [ka]

[0175] A solution was prepared by dissolving N-(3,4-dichlorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide (0.210 g, 0.636 mmol) prepared in Step 3 and sodium hydride (60.00%, 0.028 g mmol) in N,N-dimethylformamide (4 mL) at 0°C, and then adding methyl 4-(bromomethyl)-3-fluorobenzoic acid. (0.173 g, 0.700 mmol) was added and the mixture was stirred at room temperature for 18 hours. The solvent was removed from the reaction mixture under reduced pressure, and water was added to the concentrate. After extraction with dichloromethane, the mixture was filtered through a plastic filter to remove the solid residue and aqueous layer, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (SiO2, 4 g cartridge; methanol / dichloromethane = 0%~5%) and concentrated to obtain the title compound (0.170 g, 53.8%) as a pale yellow solid.

[0176] [Step 5] Synthesis of N-(3,4-dichlorophenyl)-N-(2-fluoro-4-(hydrazinecarbonyl)benzyl)-4-(oxetan-3-yl)piperazine-1-carboxamide [ka]

[0177] The methyl 4-((N-(3,4-dichlorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide)methyl)-3-fluorobenzoic acid (0.170 g, 0.342 mmol) prepared in Step 4 and hydrazine monohydrate (0.333 mL, 6.850 mmol) were dissolved in ethanol (5 mL) at room temperature. The solution was stirred at 110 °C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. After removing the solvent from the reaction mixture under reduced pressure, the title compound was used without further purification (0.170 g, 100.0%, pale yellow solid).

[0178] [Step 6] Synthesis of Compound 4 [ka]

[0179] In Step 5, N-(3,4-dichlorophenyl)-N-(2-fluoro-4-(hydrazinecarbonyl)benzyl)-4-(oxetan-3-yl)piperazine-1-carboxamide (0.170 g, 0.342 mmol) and imidazole (0.070 g, 1.027 mmol) were prepared by dissolving them in dichloromethane (4 mL) at room temperature. To this solution, 2,2-difluoroacetic anhydride (0.128 mL, 1.027 mmol) was added, and the mixture was heated under reflux for 18 hours. After cooling to room temperature, the reaction was terminated. After removing the solvent from the reaction mixture under reduced pressure, the concentrate was purified by column chromatography (SiO2, 4 g cartridge; methanol / dichloromethane = 0%~2.5%) and concentrated to obtain the desired compound 4 (0.020 g, 10.5%) as a white solid.

[0180] 1 H NMR (400 MHz, CDCl3) δ 7.89 (d, J = 7.8 H z, 1H), 7.78 (d, J = 9.9 Hz, 1H), 7.68 (t, J = 7.3 Hz, 1H), 7.38 (d, J = 8.6 Hz, 1H), 7.23 (s, 1H), 7.06 - 6.80 (m, 2H), 4.95 (s, 2H), 4.64 (t, J = 6.1 Hz, 2H), 4.55 (t, J = 5.4 Hz, 2 H), 3.45 - 3.42 (m, 1H), 3.34 - 3.33 (m, 4H), 2.18 - 2.17 (m, 4H); LRMS (ES) m / z 556.2 (M + + 1).

[0181] Examples 5-8: Synthesis of Compounds 5-8 In Step 3 of Example 4, instead of 3,4-dichlorophenyl isocyanate, Reactant A in Table 2 below was used, and in Step 4, instead of methyl 4-(bromomethyl)-3-fluorobenzoate, Reactant B in Table 2 below was used. Compounds 5, 6, 7, and 8 according to Example 5, Example 6, Example 7, and Example 8 were respectively produced by the same reactions as described in Steps 3 to Step 6 of Example 4, except for the above changes. The properties and yields of the products produced in each of Steps 3 to Step 6 of Example 5 to Example 8 are shown in Table 2, and their NMR data are shown in Table 3. In Table 3 above, Compound 5, Compound 6, Compound 7, and Compound 8 are compounds produced by Example 5, Example 6, Example 7, and Example 8 respectively.

[0182]

Table 2

[0183]

Table 3

[0184] Activity Measurement and Analysis Protocol of the Compounds of the Present Invention Experimental Example 1. HDAC Enzyme Activity Inhibition Assay (in vitro) To confirm the selectivity of the compounds represented by Chemical Formula I of the present invention for HDAC6, experiments were carried out through HDAC1 and HDAC6 enzyme activity inhibition experiments.

[0185] HDAC enzyme activity was measured using the HDAC Fluorimetric Drug Discovery Kit (BML-AK511, 516) from Enzo Life Science. For the HDAC1 enzyme activity test, human recombinant HDAC1 (BML-SE456) was used as the enzyme source, and Fluor de Lys (registered trademark)-SIRT Compound 1 (BNL-KI177) was used as the substrate. After dispensing the compound, diluted 5-fold, into a 96-well plate, 0.3 μg of enzyme and 10 μM substrate were added to each well. The mixture was reacted at 30°C for 60 minutes. Then, Fluor de Lys® Developer II (BML-KI176) was added and reacted for 30 minutes. The reaction was then terminated, and fluorescence values ​​(Ex 360, Em 460) were measured using a multi-plate reader (Flexstation 3, Molecular Device). The HDAC6 enzyme was tested using Calbichem's human recombinant HDAC6 (382180) following the same protocol as the HDAC1 enzyme activity test. The final results were obtained using GraphPad Prism. Each IC using the 4.0 program 50 The value was calculated.

[0186] [Table 4]

[0187] As shown in Table 4 above, the results of activity inhibition tests against HDAC1 and HDAC6 confirmed that the 1,3,4-oxadiazole oxetane derivative compounds of the present invention, their stereoisomers, or pharmaceutically acceptable salts exhibit excellent selective HDAC6 inhibitory activity of approximately 517 to 1207 times.

[0188] Experimental Example 2. Analysis of HDAC6 inhibitory activity and selectivity for other HDAC isotypes (in vitro) The HDAC6 inhibitory effect and selectivity of the compound referred to as Compound 5 (Compound 5 in Example 5: Compound 5) and the compound referred to as Compound 1 (Compound 1 in Example 1: Compound 1) were confirmed at the enzymatic level. This experiment was commissioned to Reaction Biology Corp. (Malvern, PA, USA) and was performed according to the established test methods within the institution. Specifically, the compound referred to as Compound 5 or Compound 1 was sequentially diluted and dispensed, and then the substrate RHK-K(Ac)-AMC and the enzyme were added together to 50 mM Tris-HCl buffer (pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, 1 mg / ml BSA) to induce the reaction. Subsequently, a 50 ml Tris-HCl buffer (pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2) containing 2 mM nicotinamide and 16 mg / mL trypsin was added and allowed to react. Enzyme activity was then measured by measuring the fluorescence signal at Ex. 360 nm / Em. 460 nm, and the results are shown in Table 5 below.

[0189] [Table 5]

[0190] IC of compound 5 against HDAC6 50 The concentration was 18.9 nM, and it was confirmed that it did not inhibit any other HDAC isotypes. Compound 1 IC for HDAC6 50 The concentration was 60.0 nM, and it was confirmed that it did not inhibit any other HDAC isotypes. In other words, the compound referred to as 13608 and the compound referred to as compound 1 exhibited excellent inhibitory activity against HDAC6 and were confirmed to be compounds that showed high selectivity for HDAC6 compared to other HDAC isotypes.

[0191] Experimental Example 3. Analysis of the effect on the migration speed of mitochondria within axons (in vitro) In several degenerative brain diseases, a decrease in tubulin acetylation, a component of microtubules, has been reported. Furthermore, it is known that intracellular transport mediated by microtubules is impaired due to various other intracellular dysfunctions.

[0192] HDAC6 is an enzyme that plays a role in removing tubulin acetylation. It is known that inhibiting HDAC6 increases tubulin acetylation, stabilizing microtubules and positively impacting intracellular and axonal transport.

[0193] Compound 5 of the present invention (compound of Example 5: compound 5) and compound 1 The compound referred to as (Compound 1 in Example 1: Compound 1) selectively inhibits HDAC6 activity and increases the acetylation of tubulin, the main substrate of HDAC6. This allowed us to confirm the effect of this compound on improving mitochondrial migration speed, which is reduced by treatment with amyloid-beta protein fragments (Aβ), a causative agent of dementia, a degenerative brain disease, within the axons of nerve cells.

[0194] Specifically, hippocampal tissue from mouse fetuses obtained from ICR mouse pregnant mothers was isolated into single cells, and the hippocampal neurons were cultured for 7 days in imaging culture vessels coated with extracellular matrix. After 7 days of culture, the mouse hippocampal neurons were treated with Aβ at a concentration of 1 μM. After 24 hours, a compound referred to as compound 5 and a compound referred to as compound 1 were treated at a concentration of 0.3 μM for 3 hours. After processing, the movement of stained mitochondria was captured at low speed using an automated cell imaging system, and the distance traveled per unit time was measured to evaluate the degree of intracellular transport.

[0195] Images were taken every minute at 1-second intervals to measure the movement speed of each mitochondrion per second. After setting a section where the movement speed of mitochondria was significantly decreased in the amyloid β treatment group compared to Vehicle in the normal group, the results normalized against the Vehicle group are shown in Table 6 below.

[0196]

Table 6

[0197] In Table 6 above, Normal is the normal group treated only with Vehicle (0.5% DMSO) without treating with amyloid β (Aβ). In amyloid β, Vehicle indicates the group treated only with amyloid β and Vehicle (0.5% DMSO).

[0198] As shown in Table 6 above, the movement speed (Relative velocity) of mitochondria, which was decreased in the nerve cell group treated with Aβ compared to the normal nerve cell group, increased by treating with a compound called Compound 5 and a compound called Compound 1 for 3 hours. It was found that the decreased movement speed of mitochondria in the pathological conditions of neurodegenerative atrophy of the nervous system including central nervous system atrophy, neurodegenerative diseases (including degenerative brain diseases) or hereditary neuropathies was improved by the treatment with the compound. As a result, it was confirmed that the compound of the present invention exhibits excellent preventive and therapeutic effects against neurodegenerative atrophy of the nervous system including central nervous system atrophy, neurodegenerative diseases (including degenerative brain diseases) or hereditary neuropathies, including dementia and Alzheimer's disease.

[0199]

[0200] Experimental Example 4. Analysis of the effect on the movement speed of mitochondria in axons during treatment at each concentration (in vitro) The compound referred to as Compound 5 of the present invention (Compound 5 of Example 5: Compound 5) selectively inhibits HDAC6 activity and increases the acetylation of tubulin, the main substrate of HDAC6. This allowed for the confirmation of a concentration-dependent improvement effect on the mitochondrial migration rate, which is reduced by treatment with amyloid-beta protein fragments (Aβ), a causative agent of dementia among degenerative brain diseases, within the axons of nerve cells. The experiments of this example were conducted to confirm this effect.

[0201] Specifically, hippocampal tissue from mouse fetuses obtained from pregnant mice was isolated into single cells, and the hippocampal neurons were cultured for 7 days in imaging culture vessels coated with extracellular matrix. After 7 days of culture, the mouse hippocampal neurons were treated with Aβ at a concentration of 1 μM. After 24 hours, a compound referred to as compound 5 was treated at concentrations of 0.01, 0.03, 0.1, 0.3, 1, 3, 10, and 30 μM for 3 hours. Subsequently, the movement of stained mitochondria was captured using a confocal microscope at low speed, and the distance traveled per unit time was measured to evaluate the degree of intracellular transport.

[0202] Images were taken at 1-second intervals for 1 minute to measure the migration speed of each mitochondrion per second. After identifying a section in the amyloid-beta-treated group where the mitochondrial migration speed was significantly reduced compared to the normal group, the data was normalized by setting the normal group to 100% and the group treated with amyloid-beta alone to 0%. The results are shown in Table 7 and Figure 1 below.

[0203] [Table 7]

[0204] In Table 7 above, "Normal" refers to the normal group treated only with Vehicle (0.5% DMSO) and not with amyloid-beta (Aβ), while "Vehicle" refers to the group treated only with amyloid-beta and Vehicle (0.5% DMSO) for amyloid-beta.

[0205] As shown in Table 7 and Figure 1 above, the relative velocity of mitochondria, which was reduced in the Aβ-treated neuronal population compared to the normal neuronal population, significantly increased in a dose-dependent manner after 3 hours of treatment with a compound called compound 5. This indicates that the reduced mitochondrial migration velocity in neuronal system atrophy, including central nervous system system atrophy, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy was improved by treatment with the compound.

[0206] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including central nervous system system atrophy, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy, including dementia and Alzheimer's disease.

[0207] Experimental Example 5. Analysis of the effect of drug efficacy on the migration speed of mitochondria within axons (in vitro) The experiment of this example was conducted to confirm the duration of the improvement effect of the compound referred to as Compound 5 of the present invention (Compound of Example 5: Compound 5) on the mitochondrial migration rate, which is reduced by treatment with amyloid-beta protein fragments (Aβ).

[0208] Specifically, hippocampal tissue from mouse fetuses obtained from pregnant mice was isolated into single cells, and the hippocampal neurons were cultured for 7 days in imaging culture vessels coated with extracellular matrix. After 7 days of culture, the mouse hippocampal neurons were treated with Aβ at a concentration of 1 μM. After 24 hours... Compound 5 was applied to the cells at a concentration of 0.3 μM for 3 hours, and then the culture medium was replaced with fresh medium. Subsequently, in the untreated state, the persistence of the drug effect was confirmed over a period of 2 to 24 hours. Using a confocal microscope, the movement of stained mitochondria was captured at low speed, and the distance traveled per unit time was measured to evaluate the degree of intracellular transport.

[0209] Images were taken at 1-second intervals for 1 minute to measure the migration speed of each mitochondrion per second. After identifying a section in the amyloid-beta treated group where the mitochondrial migration speed was significantly reduced compared to the normal group's Vehicle, the data was normalized by setting the Vehicle to 100% and the amyloid-beta treated group to 0%. The results are shown in Table 8 and Figure 2 below.

[0210] [Table 8]

[0211] In Table 8 above, "Normal" refers to the normal group treated only with Vehicle (0.5% DMSO) and not with amyloid-beta (Aβ), while "Vehicle" refers to the group treated only with amyloid-beta and Vehicle (0.5% DMSO) for Aβ.

[0212] As shown in Table 8 and Figure 2 above, in the Aβ-treated nerve cell population, the mitochondrial migration velocity (Relative velocity), which was reduced compared to the normal nerve cell population, was significantly increased by treatment with a compound called compound 5 for 3 hours. Furthermore, it was confirmed that the drug effect significantly persisted for up to 9 hours even after the compound was removed by changing to a new culture medium.

[0213] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including central nervous system system atrophy, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy, including dementia and Alzheimer's disease.

[0214] Experimental Example 6. Analysis of the effect on the migration speed of damaged mitochondria within axons in a condition with symptoms similar to tauopathy (in vitro). To confirm the therapeutic effect of compound 5 (compound of Example 5: compound 5) on degenerative brain diseases, a cell model of tauopathy was prepared and treated with the compound of the present invention. Then, to confirm the duration of the improvement effect on the reduced mitochondrial migration rate, The example experiment was conducted.

[0215] Tau proteins are known to bind to microtubules that make up nerve cells, contributing to the maintenance of microtubule stability. In pathological conditions such as tauopathy, tau proteins detach from microtubules, causing microtubule instability and adversely affecting various cellular functions. Microtubules act as roads within nerve cells, mediating the movement of various organelles, vesicles, and substances required for cellular homeostasis. However, in tauopathy, this intracellular transport is impaired due to the instability of microtubules. A test system was created using primary mouse cultured cells overexpressing human tau protein with the P301L mutation, which promotes aggregation and detachment from microtubules. In this test system, we evaluated whether a compound referred to as compound 5 improved the normalization of microtubules and intracellular transport.

[0216] Specifically, cerebral cortical tissue from mouse fetuses obtained from pregnant mice was isolated into single cells, and a P301L mutant human tau protein expression vector was transduced using electroporation. After culturing in a cell culture machine for 7 days, the cells were treated with a compound referred to as compound 5 at concentrations of 0.01, 0.1, 0.3, 1, 3, and 10 μM for 3 hours. Subsequently, the movement of stained mitochondria was captured using a confocal microscope at low speed, and the distance traveled per unit time was measured to evaluate the degree of intracellular transport.

[0217] All results are presented as mean ± standard error, and the efficacy of the drug was determined by the statistical significance between the negative control group and each test substance group. For statistical analysis, homogeneity of variance was confirmed using one-way ANOVA, and a Dunnett post-test was performed. A p-value of less than 0.05 was considered statistically significant, and the test results are shown in Table 9 and Figure 3 below.

[0218] [Table 9]

[0219] In Table 9 above, P301L_Tau(-) refers to a group of normal nerve cells transduced with a control vector that does not cause tauopathy, and treated only with vehicle (0.5% DMSO), while P301L_Tau(+) refers to a group of diseased nerve cells transduced with a P301L mutant human tau protein expression vector, and treated only with vehicle (0.5% DMSO).

[0220] As shown in Table 9 and Figure 3, in the P301L_Tau(+) group, the relative velocity of mitochondria, which was decreased compared to the P301L_Tau(-) group, significantly increased in a dose-dependent manner after 3 hours of treatment with a compound called compound 5, indicating the decreased mitochondrial movement in the pathological state of tauopathy. It was found that the dynamic speed was improved by treatment with the compound of the present invention.

[0221] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including tauopathy, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy.

[0222] Experimental Example 7. Analysis of the inhibitory activity and selectivity of HDAC6 at the neuronal level (in vitro) Compound 5 (Compound of Example 5: Compound 5) and Compound 1 The HDAC6 inhibitory effect and selectivity of the compound (compound from Example 1: Compound 1) were confirmed at the neuronal level.

[0223] The experiments of this embodiment were conducted to confirm that selective inhibition of HDAC6 activity in nerve cells by the compounds represented by compound 5 and compound 1 of the present invention increases the acetylation of tubulin, the major substrate of HDAC6.

[0224] Specifically, SH-SY5Y cells, a type of human neuroblastoma, were attached to a culture dish and cultured in an incubator for 24 hours. After 1 day of culture, the SH-SY5Y cells were treated with a compound referred to as compound 5 and a compound referred to as compound 1 at concentrations of 0.1, 0.3, and 1 μM for 3 hours. After chemical disruption using RIPA buffer, the extracted proteins were arranged by protein size on a polyacrylamide gel using SDS-PAGE. The proteins on the gel were then transferred to a nitrocellulose membrane using electrophoresis, and antibodies that selectively bind to the target protein were reacted with the membrane. The expression level was measured by visualizing the result as a band, and the results are shown in Figures 4 and 5.

[0225] In Figures 4 and 5, con represents the result for cells that have not been treated with the compound of the present invention.

[0226] As can be seen in Figures 4 and 5 above, the compound referred to as compound 5 and the compound referred to as compound 1 The compound was found to significantly increase the acetylation of tubulin.

[0227] Therefore, it can be seen that the compound according to the present invention exhibits an excellent effect in increasing tubulin acetylation.

[0228] Experimental Example 8. Analysis of brain permeability of HDAC6-specific inhibitors (in vivo) To investigate the therapeutic effects of compounds in degenerative brain diseases, the compounds exposed to the blood must be able to cross the brain-blood barrier (BBB) ​​and reach nerve cells within the brain tissue.

[0229] Compound 5 of the present invention (compound of Example 5: compound 5) and compound 1 The experiment described in this example was conducted to confirm that when the compound referred to as (compound of Example 1: compound 1) is administered orally, it is absorbed in the stomach, crosses the blood-brain barrier, and reaches nerve cells in brain tissue.

[0230] Specifically, ICR mice were orally administered a single dose of a compound referred to as compound 5 at a concentration of 10 mg / kg. Blood and brain samples were collected after 0.5, 2, and 4 hours. The residual concentration of compound 5 was measured in the plasma of the collected mouse blood and brain tissue using LC-MS / MS (see Rapid Commun. Mass Spectrom. 14, 1729-1735 (2000)). ICR mice were also given a dose of a compound referred to as compound 1 at a concentration of 50 mg. A single oral dose was administered at a concentration of / kg, and blood and brain samples were collected after 0.5, 2, and 4 hours. In the collected mouse blood plasma and brain tissue, compound 1 was identified by LC-MS / MS. The residual concentrations of the compounds were measured, and the results are shown in Table 10.

[0231] All results are expressed as mean ± standard deviation of the mean. The concentration of compounds in the brain is in ng / g, and the concentration of compounds in plasma is in ng / mL.

[0232] [Table 10]

[0233] As shown in Table 10 above, the compound referred to as Compound 5 and the compound referred to as Compound 1 are, It exhibited a B / P ratio of 0.3 or higher, confirming that it possesses the characteristic of high brain permeability.

[0234] Therefore, it can be seen that the compounds according to the present invention exhibit excellent brain permeability and show excellent preventive and therapeutic effects against neurological diseases such as brain diseases, central nervous system atrophy, or hereditary neurological diseases.

[0235] Experiment Example 9. Assessment of animal behavior (assessment of cognitive function) Cognitive impairment is the most common symptom of degenerative neurological diseases, and various assessment tools are used in clinical practice to evaluate patients' cognitive function (e.g., MoCA, MMSE, CDR, ADAS-cog). In animal experiments, cognition / learning assessment methods mainly include the Y-maze, manual avoidance test, and underwater maze test.

[0236] To confirm the therapeutic effect of the drug on cognitive impairment caused by neurodegeneration, we examined its efficacy in tauopathy mice (PS19) that develop the disease through transfection with tau, the causative protein of Alzheimer's disease and tauopathy.

[0237] Tauopathy is one of the most common degenerative brain diseases, and its causative protein, tau, is known to be involved in the development of at least 10 different degenerative brain diseases. It is a disease that develops in people who have various mutations in MAPT, the gene responsible for tauopathy.

[0238] Tau proteins are known to bind to microtubules that make up nerve cells, contributing to the maintenance of microtubule stability. In pathological conditions such as tauopathy, tau proteins detach from microtubules, causing them to become unstable and negatively impacting various cellular functions.

[0239] The PS19 tauopathy mouse is a disease model animal that overexpresses the human P301S mutant tau protein, which promotes tau protein aggregation and detachment from microtubules, resulting in increased tau and phosphorylated tau proteins in the brain, impaired memory and cognitive function, and impaired motor function, among other clinical symptoms of tauopathy.

[0240] The compound of the present invention was orally administered twice daily to PS19 tauopathy mice, and the test substance was administered. The following behavioral test was performed 30 minutes later.

[0241] Y-Maze Test As an experiment to measure short-term memory, a Y-maze test was conducted to evaluate the ability to act sequentially. The measuring device consisted of three arms, each 42 cm long, 3 cm wide, and 12 cm high, positioned at a 120° angle. The device was made of white polyvinyl plastic, and the three arms were labeled A, B, and C before the experiment. After placing the experimental animals inside for 8 minutes, points were awarded for each arm, based on the number of times the animal entered the arm up to its tail and the number of times it entered each arm in sequence (actual alternation). Alternation behavior was defined as entering all three arms without overlapping, and spontaneous alternation was calculated using the following formula, the results of which are shown in Figures 6 to 8.

[0242] In Figures 6 to 8, WT (Littermate) refers to a normal group of mice orally administered 0.5% methylcellulose, PS19 (vehicle Littermate) refers to PS19 mice (control group) orally administered 0.5% methylcellulose, and PS19 + compound 5 refers to the group of PS19 mice orally administered a compound called compound 5.

[0243] As can be seen in Figure 6, repeated administration (oral administration) of compound 5 (compound from Example 5: compound 5) for 8 weeks to 12-week-old tauopathy (PS19) mice resulted in a significant increase in alternating behavior values ​​compared to the control group (Littermate (Vehicle PS19) administered 0.5% methylcellulose), indicating an improvement in cognitive decline due to tauopathy.

[0244] As can be seen in Figure 7, repeated administration (oral administration) of compound 5 (compound from Example 5: compound 5) to 12-week-old tauopathy (PS19) mice for 16 weeks resulted in increased alternating behavior values ​​and improved cognitive function due to tauopathy compared to the control group (Littermate (Vehicle PS19) administered 0.5% methylcellulose).

[0245] As can be seen in Figure 8, compound 1 was administered to 28-week-old tauopathy (PS19) mice. The compound in Example 1 (compound 1) was administered orally for eight weeks, and it was found that alternating behavioral values ​​increased and cognitive decline due to tauopathy improved compared to the control group (Littermate (Vehicle PS19) administered 0.5% methylcellulose).

[0246] Therefore, it can be seen that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including tauopathy, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy.

[0247] Experimental Example 10. Histopathological Analysis Similar to Experimental Example 9, the test substance was administered to PS19 mice, and after the administration of the substance, histopathological analysis of brain tissue was performed. Final results for Compound 5 (Compound of Example 5: Compound 5) 0.5 hours after administration, the mice were anesthetized with isoflurane, and their brains were removed. For double immunofluorescence staining, brain tissue sections were washed twice with phosphate-buffered saline (PBS) for 15 minutes each, and then reacted with a blocking solution of bovine serum albumin (Sigma, USA) mixed with PBS for 2 hours. After that, the sections were washed twice with PBS, and the primary antibody was Triton x-100 (0.3%). The samples were diluted with PBS containing 5% normal donkey serum and reacted overnight at 4°C. Primary antibodies used included anti-phospho-Tau (Ser202, Thr205), monoclonal antibody (clone AT8, 1:300, Invitrogen, MN1020), and anti-NeuN polyclonal antibody (1:300, Invitrogen, PA5-78499). Subsequently, the tissue was washed with PBS and reacted with the secondary antibodies Alexa488-tagged donkey anti-mouse IgG (1:500, Jackson Immunoresearch, 715-545-150, for phospho-tau) and Cy3-tagged donkey anti-mouse IgG (1:500, Jackson Immunoresearch, 711-165-152, for NeuN) at room temperature for 3 hours. After re-washing with PBS, the tissue was attached to a coated slide, thoroughly dried, and then mounted in a DAPI-containing mounting medium (VECTASHIELD® Antifade Mounting Medium with DAPI, H-1200-10, Vector Laboratories) to prepare tissue specimens.

[0248] Brain tissue stained using a confocal microscope was imaged, and the staining intensity of AT8 in each brain tissue region was measured using the imageJ program. The results are shown in Figures 9 to 13.

[0249] In Figures 9 to 13, WT (Littermate) refers to a normal group of mice administered 0.5% methylcellulose, PS19 (vehicle Littermate) refers to PS19 mice (control group) administered 0.5% methylcellulose, and PS19 + compound 5 refers to the group of PS19 mice administered a compound called compound 5.

[0250] As can be seen in Figures 9-13, after administering a compound called compound 5 to 12-week-old tauopathy (PS19) mice repeatedly for 8 weeks, histopathological analysis of brain regions of the mice showed that the control group (vehicle LitterMate) mice had significantly higher levels of transfected tau hyperphosphorylation (AT8, S202 / T205) compared to the normal group (WT LitterMate) mice. On the other hand, animals administered with compound 5 showed a significant improvement in tau hyperphosphorylation compared to the control group mice.

[0251] This indicates that the compound referred to as compound 5 improves the tau hyperphosphorylation observed in PS19 mice.

[0252] Therefore, it can be seen that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including tauopathy, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy.

[0253] Experimental Example 11. Electrophysiological Analysis Similar to Experimental Example 9, the test substance was administered to PS19 mice, and electrophysiological analysis was performed on the brain tissue after the completion of substance administration. Final administration of Compound 5 (Compound 5 of Example 5: Compound 5) After 0.5 hours had passed since administration, the mice were anesthetized and their brains were removed. The removed brains were then sliced ​​into 300 μm thick sections to prepare tissue sections containing the hippocampus, and these sections were transferred to a recording chamber perfused with artificial cerebrospinal fluid (30°C-32°C) for further investigation.

[0254] fEPSP (field excitatory postsynaptic potential) recordings were performed using a DAM80 amplifier and WinLTP 2.10 software (University of Bristol), with filtering at 1 kHz and sampling at 20 kHz. Recording pipettes with resistances of 1–3 MΩ were filled with artificial cerebrospinal fluid. All fEPSPs were recorded in the Schaffer collateral pathway (CA3 to CA1 synapse) by stimulation using two bipolar electrical stimulation electrodes (FHC) placed in the radiating layer, or by stimulation with a 20-second interval (ISI). Theta burst stimulation (TBS) was applied with 100 Hz (5 pulses, 20 times) at 5 Hz intervals. Successful LTP induction was confirmed by statistical comparison of the average fEPSP gradient / amplitude measured 50–60 minutes after TBS with the average fEPSP gradient / amplitude measured 10 minutes before TBS. WinLTP 2.10 reanalysis software (University of Bristol) was used for data analysis. All data are presented as mean ± standard error of the mean (SEM), and statistical analysis was performed using SPSS Statistics version 21 (IBM). Student's t-test was used for significance testing when comparing two groups, and ANOVA was used for significance testing for group comparisons. Post-hoc LSD analysis was performed, and the results are shown in Figures 14 and 15.

[0255] The significance level was set at p<0.05. * or § means p<0.01; ** or §§ means p<0.01; *** or §§§ means p<0.001.

[0256] In Figure 15, in 7 / 2, 7 represents the number of brain tissue slices and 2 represents the number of mice evaluated. In 14 / 4, 14 represents the number of brain tissue slices and 4 represents the number of mice evaluated.

[0257] In Figures 14 and 15, WT (Littermate) refers to normal mice orally administered 0.5% methylcellulose, PS19 (vehicle Littermate) refers to PS19 mice (control group) orally administered 0.5% methylcellulose, and PS19 + compound 5 refers to the group of PS19 mice orally administered compound 5. As confirmed in Figures 4 and 15, the magnitude of long-term memory enhancement (LTP) in the hippocampus of the control group was significantly reduced compared to the normal group. This indicates a deterioration of the neurological pathways associated with memory. On the other hand, repeated oral administration of a compound called compound 5 to 12-week-old tauopathy (PS19) mice for 16 weeks resulted in a significant recovery of hippocampal long-term memory enhancement compared to the control group (Littermate (vehicle PS19) administered 0.5% methylcellulose).

[0258] Therefore, it can be seen that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including tauopathy, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy.

[0259] Experimental Example 12. Assessment of animal behavior (assessment of motor function) To confirm the therapeutic effects of the compound referred to as 13524 (compound 1 in Example 1) and the compound referred to as compound 5 (compound 5 in Example 5), which are used to treat degenerative brain diseases, the compounds were administered to Yac128 mice, a Huntington's disease model mouse, and the improvement in the animals' motor function was evaluated.

[0260] Huntington's disease is one of the most common degenerative brain disorders, a genetic disorder that leads to severe physical and mental impairment within 15 to 25 years of onset, and often results in death. It is an autosomal dominant disorder that develops in individuals with a mutation in the HTT gene, the causative gene for Huntington's disease, in which the CAG sequence is repeated 40 or more times.

[0261] Yac128 is a Huntington's disease model mouse in which a human mutant HTT gene with 128 repeats of the CAG sequence is inserted. It expresses the human mutant HTT transcript and protein, and is a disease model animal that exhibits clinical symptoms of Huntington's disease, such as striatal death and decreased motor function, including muscle strength and limb coordination.

[0262] Specifically, six-month-old Yac128 mice were given a compound called compound 1 in a 5:20 ml solution. The compound referred to as Compound 5 was administered orally at g / kg twice daily for 8 weeks, and at 1, 3, 10, and 30 mg / kg twice daily for 12 weeks. Motor function was evaluated at 4-week intervals, and the results are shown in Figures 16 to 19.

[0263] In Figures 16 to 19, all results are shown as mean ± standard error, and the efficacy of the drug was determined by the statistical significance between the Yac128 mouse control group and each test substance group. Statistical analysis was performed using ANOVA to confirm the homogeneity of the variance (on-way ANOVA for single measurements, two-way ANOVA for repeated measurements), followed by a Dunnett or Bonferroni post-test. A p-value of less than 0.05 was considered statistically significant. Accelerating rotarod and grip strength tests were performed once daily for two days. Subsequently, based on the test results, groups were separated using the Z-array method. Yac128 mice were divided into a vehicle administration group and a compound administration group (compound 1 and The mice were divided into groups of 5 (20 mg / kg) and 1, 3, 10, and 30 mg / kg (oral administration), with 18 mice in each group. The test substance was administered orally twice a day for 8 and 12 weeks, and motor function was evaluated once a day for two days at 4-week intervals during the administration period.

[0264] Accelerator rotor rod test To evaluate motor coordination and motor function, an accelerated rotorod test (ROTA ROD, LE8205, Panlab) was performed. Prior to the test, all test animals underwent adaptation training for approximately three weeks, placing them on a rod accelerating from 4 to 20 rpm three times a day for three days. Animals that fell from the rod for 180 seconds or more were used in the test. In this experiment, the time it took for an animal to fall from a rod accelerating from 4 to 40 rpm was measured for three minutes. The accelerated rotorod test was performed three times a day for two consecutive days, for a total of six tests, and the maximum value from the six measurements was used. The results are shown in Figures 16 and 17.

[0265] In Figures 16 and 17, WT (wild type) refers to normal mice orally administered with vehicle (0.5% methylcellulose), Yac128 refers to the Huntington's disease model mouse control group orally administered with vehicle, Yac128+ Compound 5 refers to the group of Yac128 mice orally administered with a compound called Compound 5, and Yac128+ Compound 1 refers to the group of Yac128 mice orally administered with a compound called Compound 1.

[0266] As shown in Figure 16, the latency to fall, which was reduced in the Yac128 control group compared to normal mice, was due to compound 1. The combined substance was administered at a dose of 20 mg / kg for 4 weeks, resulting in a significant increase.

[0267] Furthermore, as shown in Figure 17, the latency to fall, which was reduced in the Yac128 control group compared to normal mice, significantly increased after 1 week of administration of a compound called compound 5 at a dose of 3 mg / kg. This significant increase was observed in most cases at longer administration periods or higher concentrations, indicating that ataxia, a symptom of Huntington's disease, was improved by administration of the compound of the present invention.

[0268] Therefore, it can be seen that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Huntington's disease, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy.

[0269] Grip strength test To evaluate muscle strength, a grip strength test (GRIP STRENGTH TEST, BIO-GS3, BIOSEB) was performed. The grip strength test used a bar to assess the strength of the mouse's two forelimbs. All experiments were conducted on one mouse. After the mouse gripped the bar, its tail was gently pulled to elicit grip strength, and then the maximum tension was measured by pulling at a 15° incline. A total of 10 grip strength tests were performed, five times a day for two consecutive days. The results, using the average of these values, are shown in Figures 18 and 19.

[0270] In Figures 18 and 19, WT (wild type) refers to normal mice administered with vehicle, Yac128 refers to the control group of Huntington's disease model mice administered with vehicle, Yac128+ Compound 5 refers to the group of Yac128 mice administered with a compound called Compound 5, and Yac128+ Compound 1 refers to the group of Yac128 mice administered with a compound called Compound 1.

[0271] As shown in Figure 18, grip strength, which was reduced in the Yac128 control group compared to normal mice, significantly increased in a dose-dependent manner after 8 weeks of administration of a compound referred to as compound 1.

[0272] Furthermore, as shown in Figure 19, grip strength, which was reduced in the Yac128 control group compared to normal mice, was reduced after administering a compound called compound 5 for 12 weeks. This resulted in a significant increase, and a significant increase was observed in all dose concentrations, indicating that muscle weakness, a symptom of Huntington's disease, was improved by administering the compound of the present invention.

[0273] Therefore, it can be seen that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Huntington's disease, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy.

[0274] Experimental Example 13. Effect of increasing the migration speed of damaged mitochondria within axons in Charcot-Marie-Tooth disease (CMT, HMSN, hereditary motor-sensory neuropathy) (in vitro) To confirm the therapeutic effect of the compound referred to as Compound 5 (Compound of Example 5: Compound 5) and the compound referred to as Compound 1 (Compound of Example 1: Compound 1) on CMT disease, the experiment of this example was conducted to confirm the improvement effect obtained by treating the compound of the present invention with the reduced mitochondrial motility in the axons of neurons isolated from the dorsal root ganglia of MFN2 mutant mice (9 months old) in which CMT2A type Charcot-Marie-Tooth disease was induced.

[0275] Mouse neurons obtained by isolating the dosal root ganglia (DRG) from 9-month-old MFN2 mutant mice were cultured in a cell culture machine for 3 days, and then treated with compounds referred to as compound 5 and compound 1 at concentrations of 100 and 300 nM for 3 hours. Subsequently, the movement of stained mitochondria was captured using a confocal microscope at low speed, and the distance traveled per unit time was measured to evaluate the degree of intracellular transport.

[0276] Results are shown as mean ± standard error, and the efficacy of the drug is compared between the negative control group and each test substance group. The determination was based on statistical significance. For statistical analysis, homogeneity of variances was confirmed using one-way ANOVA, and a Dunnett post-test was performed. If the p-value was less than 0.05, it was considered statistically significant.

[0277] Also, MFN R94QIn mouse DRG, after identifying a compartment in which mitochondrial migration rate was significantly reduced compared to the normal group (WT DRG), the normal group (100%) and the MFN group (100%) were separated. R94Q The mouse DRG was normalized to 0%, and the test results are shown in Table 11 and Figure 20 below.

[0278] In Table 11 above, WT refers to the group of normal mice administered with vehicle (0.5% methylcellulose), and MFN refers to the group of normal mice. R94Q The mouse DRG group consisted of MFN2 mutant mice (inducible CMT2A type Charcot-Marie-Tooth disease) that were administered only Vehicle (0.5% methylcellulose).

[0279] [Table 11]

[0280] As shown in Table 11 and Figure 20 above, in the MFN2 mutant control group, the relative velocity of mitochondria, which was reduced compared to the WT group, significantly increased after 3 hours of treatment with compounds referred to as Compound 1 and Compound 5. This indicates that the reduced mitochondrial migration velocity in the CMT pathology was improved by treatment with the compounds of the present invention.

[0281] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including Charcot-Marie-Tooth disease, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy.

[0282] Experimental Example 14. Confirmation of the therapeutic effect on CMT. CMT is the most common hereditary peripheral neuropathy, caused by mutations in proteins that make up nerves. To date, more than 1,000 mutations have been identified in over 90 genes (Timmerman et al., (2014) Genes 5:1). 3-32). When Charcot-Marie-Tooth disease develops, progressive degeneration of peripheral nerves causes atrophy of muscles affected by nerve distribution, resulting in gradual muscle atrophy and weakness in the limbs, and deformities of the feet and fingers. CMT is known to be highly diverse and complex both genetically and clinically, and its symptoms vary from mild, near-normal conditions to severe conditions that necessitate wheelchair use, depending on the type of mutation. It mainly develops in teenagers and occurs in approximately 1 in 2500 people (Krajewski et al., (2000) Brain 123:1516).

[0283] CMT is a rare hereditary peripheral nerve disorder. However, its prevalence is approximately 1 in 2,500 people, with about 20,000 patients in South Korea and 2.8 million worldwide. Currently, CMT is treated with limited therapies, including rehabilitation, assistive devices, pain management, and surgery. Since no effective treatment has yet been developed, the development of CMT treatments is extremely important.

[0284] For example, regarding CMT, the most common type of hereditary sensorimotor neuropathy, a large-scale clinical trial was conducted on ascorbic acid, which was proven to be an essential substance for myelin formation in the peripheral nervous system through experiments culturing Schwann cells and dorsal root ganglion cells together, but it failed to demonstrate efficacy (Pareyson et al., (2011) 10(4):3205). In particular, certain types of diseases such as CMT1X and CMT2A have high expression of the causative genes in the central nervous system, and in fact, approximately 10% of patients exhibit symptoms such as atrophy of specific brain tissues, including the optic nerve, and visual impairment, highlighting the importance of drug action in the central nervous system.

[0285] As described above, the compounds of the present invention can cross the blood-brain barrier and exhibit therapeutic effects not only against peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT), but also against central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT).

[0286] Therefore, in the following two types of CMT disease model studies, it was confirmed that the compounds of the present invention can be effectively used for the prevention and treatment of CMT by significantly improving the motor and sensory functions (rotarod and balance beam tests) of CMT mice.

[0287] 14.1 Assessment of motor and sensory functions

[0288] Neurological damage caused by hereditary neurological disorders such as Charcot-Marie-Tooth disease (CMT) results in gait disturbances, sensory loss, and impaired limb coordination. Clinical studies involving patients use various assessment tools to evaluate these functional impairments, including a 6-meter walk, sensory function assessment based on the position of arm and leg parts, and buttoning tasks. In animal experiments, methods such as the Rotarod test, balance beam test, and grip strength test are primarily used to assess symptoms resulting from such neurological damage.

[0289] In this study, the compound referred to as Compound 5 (Compound of Example 5: Compound 5) was used to treat CMT disease. To confirm the therapeutic effects on motor and sensory functions in patients, the compound was administered to two types of CMT disease model mice (CX32 null mice and MFN2 mutant mice), and the improvement in the relevant functions of the animals was evaluated.

[0290] 14.1.1 Research results of CX32 null mice First, CX32 null mice, at 5 months of age, were given a compound called compound 5 for 5 months. The drug was administered orally twice daily at a dose of 0 mg / kg, and motor and sensory functions were evaluated during the administration period. The results are shown in Figures 21 and 22.

[0291] In Figures 21 and 22 below, all results are shown as mean ± standard error, and the efficacy of the drug was determined by the statistical significance between the normal group (WT) and each test substance group. For statistical analysis, homogeneity of variance was confirmed using ANOVA (on-way ANOVA for single measurements, two-way ANOVA for repeated measurements), and a Dunnett or Bonferroni post-test was performed. A p-value of less than 0.05 was considered statistically significant.

[0292] In this study, animals were classified into groups using the Z-array method, as shown in Table 12 below, based on their body weight before drug administration, constant rotarod, and balance beam test results. Mice were orally administered either vehicle (0.5% methylcellulose) or a compound referred to as compound 5 at 10 mg / kg, depending on their assigned group.

[0293] The male CX32 null mice used in this study were provided with a standard diet (Central Lab Animal, Inc.) and water freely (ad libitum), and were housed in a controlled environment with temperature (22±2℃), humidity (44~56%), and a 12-hour dark / light cycle. All experimental procedures were carried out with approval from the IACUC (Institutional Animal Care and Use Committee) of the Korea CKD Laboratory Animal Center (Approval Number: S-22_016).

[0294] [Table 12]

[0295] Constant velocity rotor rod test The Rota Rod test (LE 8205, Panlab) was performed to evaluate forced motor activity and coordination function. For adaptation, all test animals underwent a 3-day course of adaptation training at 8 rpm five times a day, followed by two days of adaptation training at 12 rpm five times a day. Animals meeting the 100-180 second latency to fall category were used for further experiments (approximately 80% of the animals met this category). Latency to fall was measured three times at a fixed speed of 12 rpm for 3 minutes. The Rota Rod test was repeated three times per experiment, and the maximum of the three measurements was used as the test result (latency to fall).

[0296] In Figure 21, the normal group (wild type, WT) consists of normal mice orally administered with vehicle, the CX32 null mice represent the control group of CMT disease model mice orally administered with vehicle, and the CX32 null mice + compound 5 represent CMT disease mice in which CX32 is knocked out (CX32 null mice This refers to the group that was orally administered a compound called compound 5.

[0297] As a result, as shown in Figure 21, it was confirmed that the compound of the present invention significantly improves the waiting time until dropping in CX32 null mice.

[0298] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against Charcot-Marie-Tooth disease, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT), as well as neurodegenerative diseases (including degenerative brain diseases) or hereditary neuropathy.

[0299] Balance beam test The balance beam test was performed to measure motor coordination and proprioception in the limbs. A beam (1.2 cm wide, 0.6 cm high, 1.0 m long) was fixed at a 9° incline (45 cm above the starting point and 60 cm above the arrival point). At the starting point, the mouse was stimulated with 60 W of light, and at the arrival point, a dark box without light was prepared to make the mouse feel safe. All experimental animals were adapted to the same conditions as the experiment 30 minutes prior to evaluation. Mice were placed at the starting point and asked to walk to the arrival point, and the number of slips after starting was measured. Prior to the test, the mice were trained three times a day for two days, and the results from the third day were used for grouping. Each experiment was independently evaluated by two individuals, and the results are shown in Figure 22.

[0300] In Figure 22, the normal group (wild type, WT) consists of normal mice orally administered with vehicle, Veh represents the control group of CMT disease model mice orally administered with vehicle to CX32 null mice, and compound 5 represents CX32 knocked. Compound 5 was found in CMT disease mice that were eliminated (CX32 null mice). This refers to the group that received the compound orally.

[0301] As shown in Figure 22, it was confirmed that the compound of the present invention has the effect of significantly reducing the slip count and beam transverse time.

[0302] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against Charcot-Marie-Tooth disease, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT), as well as neurodegenerative diseases (including degenerative brain diseases) or hereditary neuropathy.

[0303] 14.1.2 Research results of MFN2 mutant mice Six-month-old MFN2 mutant mice were given a compound called Compound 5 at a dose of 10 mg / kg for three months. The drug was administered orally twice daily, and motor and sensory functions were evaluated during the administration period. The results are shown in Figures 23 and 24. The presentation and statistical processing methods of the results were the same as those used in the CX32 null mouse study described in 14.1.1 above.

[0304] In this study, animals were classified into groups using the Z-array method, based on their body weight before drug administration, values ​​obtained from accelerated rotarod and balance beam tests, as shown in Table 13 below. Mice were orally administered either vehicle (0.5% methylcellulose) or compound 5 at 10 mg / kg, depending on their assigned group.

[0305] The male MFN2 mutant mice used in this study were fed a standard diet (Central Lab The animals were provided with free access to water (ad libitum) and reared in a controlled environment with temperature (22±2℃), humidity (44~56%), and a 12-hour dark / light cycle. All experimental procedures were carried out with approval from the IACUC (Institutional Animal Care and Use Committee) of the Korea CKD Laboratory Animal Center (Approval Number: S-22_012).

[0306] [Table 13]

[0307] Accelerator rotor rod test To evaluate motor coordination and motor function, the accelerated rotorod test (ROTA ROD, LE8205, Panlab) was performed. Prior to the test, all test animals underwent adaptation training for approximately three weeks, placing them on a rod accelerating from 4 to 20 rpm three times a day for three days. Animals that fell from the rod for 180 seconds or more were used in the test. In this experiment, the time it took for an animal to fall from a rod accelerating from 4 to 40 rpm was measured over a three-minute period. The accelerated rotorod test was performed three times a day, and the maximum value of the three measurements was used. The results are shown in Figure 23.

[0308] In Figure 23, the normal group (wild type, WT) consists of normal mice orally administered with vehicle, while Veh represents the control group of CMT disease model mice in which MFN2 mutant mice were orally administered with vehicle (0.5% methylcellulose), and compound 5 This refers to the group of MFN2 mutant mice that were orally administered a compound called compound 5.

[0309] As a result, as shown in Figure 23, it was confirmed that the compound of the present invention has the effect of significantly improving the latency to fall.

[0310] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against Charcot-Marie-Tooth disease, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT), as well as neurodegenerative diseases (including degenerative brain diseases) or hereditary neuropathy.

[0311] Balance beam test The balance beam test was performed to measure motor coordination and proprioception in the limbs. The research method was the same as that used in the CX32 null mouse study described in 14.1.1 above, and the results are shown in Figure 24.

[0312] In Figure 24, the normal group (wild type, WT) consists of normal mice orally administered with vehicle, while Veh represents the control group of CMT disease model mice in which MFN2 mutant mice were orally administered with vehicle (0.5% methylcellulose), and compound 5 This refers to the group of MFN2 mutant mice that were orally administered a compound called compound 5.

[0313] As shown in Figure 24, it was confirmed that the compound of the present invention has the effect of significantly reducing the number of slips.

[0314] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against Charcot-Marie-Tooth disease, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT), as well as neurodegenerative diseases (including degenerative brain diseases) or hereditary neuropathy.

[0315] 14.1.3 Research results of CMT2A mice 6-month-old CMT2A mice (mutant Mfn2 R94Q Compound 5 from Example 5 was orally administered to the mice at a dose of 10 mg / kg twice daily for 3 months, and motor and sensory functions were evaluated during the administration period. The results are shown in Figures 25-26. The presentation and statistical processing of the results were carried out in the same manner as in the CX32null mouse study described in 14.1.1 above.

[0316] In this study, the animals were classified into groups using the Z-array method based on their body weight before drug administration, values ​​obtained from accelerated rotarod and balance beam tests, as shown in Table 14 below. Mice were orally administered either vehicle (0.5% methylcellulose) or 10 mg / kg of compound 5 from Example 5, depending on their assigned group.

[0317] The male CMT2A mice used in this study were provided with a standard diet (Central Lab Animal, Inc.) and water freely (ad libitum), and were housed in a controlled environment with controlled temperature (22±2℃), humidity (44~56%), and a 12-hour contrast cycle. All experimental procedures were carried out with approval from the IACUC (Institutional Animal Care and Use Committee) of the Korea CKD Laboratory Animal Center (Approval Number: S-22_012).

[0318] [Table 14]

[0319] Accelerator rotor rod test To evaluate motor coordination and motor skills, the accelerated rotarod test (ROTA ROD, LE8205, Panlab) was performed. Prior to the test, all test animals underwent adaptation training for approximately three weeks, placing them on a rod accelerating from 4 to 20 rpm three times a day for three days. Animals that fell from the rod for 180 seconds or more were used in the test. In this experiment, the time it took for an animal to fall from a rod accelerating from 4 to 40 rpm was measured over a three-minute period. The accelerated rotarod test was performed three times a day, and the maximum value of the three measurements was used. The accelerated rotarod test was conducted for three months, and the AUC data (period × measurement value in the rotarod test) obtained using the prism file is shown in Figure 25.

[0320] In Figure 25 above, the normal group (wild type, WT) took the vehicle orally. These are normal mice that were administered the compound. Veh refers to the control group of CMT disease model mice that were orally administered vehicle (0.5% methylcellulose) to CMT2A mice, and Compound 5 refers to the group of CMT2A mutant mice that were orally administered Compound 5 (Example 5).

[0321] As a result, as shown in Figure 25, it was confirmed that the compound of the present invention (for example, compound 5 of Example 5) exhibits the effect of significantly improving the latency to fall.

[0322] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT).

[0323] Balance beam test The balance beam test was performed to measure motor coordination ability and proprioception for the limbs. The research method was the same as in the CX32null mouse study described in 14.1.1 above, and the results are shown in Figure 26.

[0324] In Figure 26 above, the normal group (wild type, WT) refers to normal mice orally administered with vehicle, Veh(TG) refers to the control group of CMT disease model mice orally administered with vehicle (0.5% methylcellulose) to CMT2A mice, and Compound 5(TG) refers to the group of CMT2A mutant mice orally administered with Compound 5 (Example 5).

[0325] As shown in Figure 26 above, it was confirmed that the compound of the present invention (for example, compound 5 of Example 5) exhibits the effect of significantly reducing the slip count.

[0326] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT).

[0327] 14.1.4 Research results of CMT1X mice Five-month-old CMT1X mice (Gjb1 KO) were orally administered compound 5 from Example 5 at a dose of 10 mg / kg twice daily for six months, and motor and sensory functions were evaluated during the administration period. The results are shown in Figures 27-28. The presentation and statistical analysis of the results were carried out in the same manner as in the study of CX32null mice described in 14.1.1 above.

[0328] In this study, the animals were classified into groups using the Z-array method based on their body weight before drug administration, values ​​obtained from accelerated rotarod and balance beam tests, as shown in Table 15 below. Mice were orally administered either vehicle (0.5% methylcellulose) or 10 mg / kg of compound 5 from Example 5, depending on their assigned group.

[0329] The male CMT1X mice used in this study were provided with a standard diet (Central Lab Animal, Inc.) and water freely (ad libitum), and were housed in a controlled environment with controlled temperature (22±2℃), humidity (44~56%), and a 12-hour contrast cycle. All experimental procedures were performed at the IACUC (Insti) of the Korea CKD Laboratory Animal Center. This was carried out with approval in accordance with the National Animal Care and Use Committee (Approval Number: S-22_012).

[0330] [Table 15]

[0331] Constant velocity rotor rod test To evaluate motor coordination and motor skills, a constant-velocity rotor rod test (ROTA ROD, LE8205, Panlab) was performed. Prior to the test, all test animals underwent adaptation training for approximately three weeks, placing them on a rod rotating at 10 rpm three times a day for three days. Animals that fell from the rod for 180 seconds or more were used in the test. In this experiment, the time an animal took to fall from the rod was measured for three minutes. The constant-velocity rotor rod test was performed three times a day, and the maximum value of the three measurements was used. The results are shown in Figure 27.

[0332] In Figure 27 above, the normal group (wild type, WT) refers to normal mice orally administered with vehicle, Veh(TG) refers to the control group of CMT disease model mice orally administered with vehicle (0.5% methylcellulose) to CMT1X mice, and Compound 5(TG) refers to the group of CMT1X mice orally administered with Compound 5 (Example 5).

[0333] As a result, as shown in Figure 27, it was confirmed that the compound of the present invention (for example, compound 5 of Example 5) exhibits the effect of significantly improving the latency to fall.

[0334] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT).

[0335] Balance beam test The balance beam test was performed to measure motor coordination ability and proprioception for the limbs. The research method was the same as in the CX32null mouse study described in 14.1.1 above, and the results are shown in Figure 28.

[0336] In Figure 28 above, the normal group (wild type, WT) refers to normal mice orally administered with vehicle, Veh(TG) refers to the control group of CMT disease model mice orally administered with vehicle (0.5% methylcellulose) to CMT1X mice, and Compound 5(TG) refers to the group of CMT1X mice orally administered with Compound 5 (Example 5).

[0337] As shown in Figure 28 above, it was confirmed that the compound of the present invention (for example, compound 5 of Example 5) exhibits the effect of significantly reducing the slip count.

[0338] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT).

[0339] 14.1.5 Research results of CMT1A mice 6.5-week-old CMT1A mutant mice (hPMP22 3-4 copies) were orally administered compound 5 from Example 5 at a dose of 10 mg / kg twice daily for 6 weeks, and motor and sensory functions were evaluated during the administration period. The results are shown in Figures 29-30. The presentation and statistical analysis of the results were carried out in the same manner as in the study of CX32null mice described in 14.1.1 above.

[0340] In this study, the animals were classified into groups using the Z-array method based on their body weight before drug administration, values ​​obtained from constant rotarod and balance beam tests, as shown in Table 16 below. Mice were orally administered either vehicle (0.5% methylcellulose) or 10 mg / kg of compound 5 from Example 5, depending on their assigned group.

[0341] The male CMT1A mutant mice used in this study were fed a standard diet (Central Lab The animals were provided with free access to water (ad libitum) and reared in a controlled environment with temperature (22±2℃), humidity (44~56%), and a 12-hour contrast cycle. All experimental procedures were carried out with approval from the IACUC (Institutional Animal Care and Use Committee) of the Korea CKD Laboratory Animal Center (Approval Number: S-22_012).

[0342] [Table 16]

[0343] Constant velocity rotor rod test To evaluate motor coordination and motor skills, a constant-velocity rotorod test (ROTA ROD, LE8205, Panlab) was performed. Prior to the test, all test animals underwent adaptation training for approximately three weeks, placing them on a rod rotating at 10 rpm three times a day for three days. Animals that fell from the rod for 180 seconds or more were used in the test. In this experiment, the time an animal took to fall from the rod was measured for three minutes. The constant-velocity rotorod test was performed three times a day, and the maximum value of the three measurements was used. The results are shown in Figure 29.

[0344] In Figure 29 above, the normal group (wild type, WT) refers to normal mice orally administered with vehicle, Veh(TG) refers to the control group of CMT disease model mice orally administered with vehicle (0.5% methylcellulose) to CMT1A mice, and Compound 5(TG) refers to the group of CMT1A mice orally administered with Compound 5 (Example 5).

[0345] As a result, as shown in Figure 29, it was confirmed that the compound of the present invention (for example, compound 5 of Example 5) exhibits the effect of significantly improving the latency to fall.

[0346] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT).

[0347] Balance beam test The balance beam test was performed to measure motor coordination ability and proprioception for the limbs. The research method was the same as in the CX32null mouse study described in 14.1.1 above, and the results are shown in Figure 30.

[0348] In Figure 30 above, the normal group (wild type, WT) refers to normal mice orally administered with vehicle, Veh(TG) refers to the control group of CMT disease model mice orally administered with vehicle (0.5% methylcellulose) to CMT1A mutant mice, and Compound 5(TG) refers to the group of CMT1A mutant mice orally administered with Compound 5 (Example 5).

[0349] As shown in Figure 30 above, it was confirmed that the compound of the present invention (for example, compound 5 of Example 5) exhibits the effect of significantly reducing the slip count.

[0350] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases (including degenerative brain diseases), or hereditary neuropathy, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT).

[0351] 14.2 Nerve conduction study (NCS) This experiment aimed to evaluate the effectiveness of the compound of the present invention by confirming its effect on the nerve conduction velocity of animals.

[0352] The animals used for nerve conduction studies were the same animals used in the study described in Section 14.1.2. Six-month-old male MFN2 mutant mice were administered the compound or vehicle for three months, after which nerve conduction studies were performed.

[0353] Electrophysiological recordings were evaluated by examining electrical conduction in the caudal nerve, which is associated with sensory nerve conduction in the peripheral nervous system (PNS), under anesthesia with isoflurane (USP Terrel, Piramal Critical Care, Inc., NDC66794-017-25) in 30% oxygen (Daehan gas) and 70% nitrogen (Daehan gas). The relevant nerve conduction studies (NCS) were performed using Nicolet Viking Quest. Sensory neuron action potential (SNAP) amplitude and sensory neuron conduction velocity (SNCV) were measured.

[0354] Data are shown as mean ± SEM. Statistical significance between the compound treatment group and the vehicle group of the present invention was analyzed using One-Way ANOVA (post-hoc analysis using Dunnett's test) to compare three or more groups. All statistical analyses were performed using GraphPad Prism (ver 9.0), and the results are shown in Figure 31.

[0355] In Figure 31, the normal group (wild type, WT) consists of normal mice orally administered with vehicle, while Veh represents the control group of CMT disease model mice in which MFN2 mutant mice were orally administered with vehicle (0.5% methylcellulose), and compound 5 This refers to the group of MFN2 mutant mice that were orally administered a compound called compound 5.

[0356] As shown in Figure 31, the compounds of the present invention significantly improve SNAP and SNCV, and it has been confirmed that they can be effectively used for the prevention and treatment of CMT by improving nerve conduction velocity.

[0357] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against Charcot-Marie-Tooth disease, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT), as well as neurodegenerative diseases (including degenerative brain diseases) or hereditary neuropathy.

[0358] 14.3 Histopathology analysis This experiment aimed to confirm the effect of the compound of the present invention on the axonal size of sciatic nerve fibers.

[0359] 14.3.1 Research results of CX32 null mice The animals used for histopathological analysis were the same animals used in the study described in Section 14.1.1. Five-month-old male CX32 null mice were administered the compound or vehicle for five months, after which histopathological analysis was performed.

[0360] At 0.5 hours after the final treatment with compound 5 (10 mg / kg) of the present invention, the ischial nerve was detected. Celluloids were collected and fixed overnight in a 2.5% glutaraldehyde solution (340855, Sigma). The fixed specimens were delivered to the Department of Pathology at Ulsan University for semitin sectioning and toluidine blue staining (T3260, Sigma).

[0361] For image analysis, the fixed samples were processed using standard methods (see Acta Neuropathologica Communications volume 7, Article number: 144 (2019), Sele et al.) to prepare 0.5 μm sections, which were then stained with toluidine blue.

[0362] Histological evaluation was performed using a light microscope. Pathological changes, including demyelination, remyelination, abnormally thin myelin, and axonal morphological changes, were examined in the sections. Finally, the axonal diameter was analyzed using imageJ software, and the results are shown in Figure 32.

[0363] In Figure 32, the normal group (wild type, WT) consists of normal mice orally administered with vehicle, and Veh represents CX32 null mice that were given vehicle. This refers to the control group of CMT disease model mice that were orally administered compound 5, in which case CX32 was knocked. Compound 5 was found in CMT disease mice that were eliminated (CX32 null mice). This refers to the group that received the compound orally.

[0364] Data are shown as mean ± SEM. Statistical significance between the compound treatment group and the vehicle group was analyzed using One-Way ANOVA (post-hoc analysis using Dunnett's test) to compare three or more groups. All statistical analyses were performed using GraphPad Prism (ver 9.0).

[0365] As shown in Figure 32, the compound of the present invention was confirmed to have a significant effect in increasing the axonal size of sciatic nerve fibers.

[0366] This confirms that the compounds of the present invention exhibit excellent preventive and therapeutic effects against Charcot-Marie-Tooth disease, including peripheral nervous system (PNS)-associated Charcot-Marie-Tooth disease (CMT) and central nervous system (CNS)-associated Charcot-Marie-Tooth disease (CMT), as well as neurodegenerative diseases (including degenerative brain diseases) or hereditary neuropathy.

Claims

1. A compound represented by the following chemical formula I, its stereoisomer, or a pharmaceutically acceptable salt thereof, 【Chemistry 1】 In the aforementioned chemical formula I, X １ ~X ４ These are, independently, N or CR ｘ And here, X １ ~X ４ It is not possible for three or more to be N at the same time, and R ｘ is -H, F, Cl, Br, or I; R １ is -CX ａ H ２ , -C(X ａ ) ２ H, or -C(X ａ ) ３ where Xa is F, Cl, Br or I; R ２ and R ３ A compound represented by chemical formula I, wherein each is independently F, Cl, Br, or I, its stereoisomer, or a pharmaceutically acceptable salt thereof.

2. X １ ~X ４ These are, independently, N or CR ｘ And here, X １ ~X ４ It is not possible for three or more to be N at the same time, and R ｘ is -H or F; R １ -CX ａ H ２ or -C(X ａ ) ２ H is H, where Xa is F or Cl; R ２ and R ３ A compound represented by chemical formula I as described in claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein each is independently F or Cl.

3. The compound represented by the aforementioned chemical formula I is represented by the following chemical formula II, 【Chemistry 2】 In the aforementioned chemical formula II, X ２ is N or CR ｘ And here, R ｘ is -H, F, Cl, Br, or I the law of nature; R １ -CX ａ H ２ , -C(X ａ ) ２ H, or -C(X ａ ) ３ And here, Xa is F, Cl, Br, or I; R ２ and R ３ A compound represented by chemical formula I according to claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein each of the elements is independently F, Cl, Br, or I.

4. The compound represented by the aforementioned chemical formula I is represented by the following chemical formulas II-1, II-2, II-3, or II-4: 【Transformation 3】 【Chemistry 4】 【Transformation 5】 【Transformation 6】 In the aforementioned chemical formulas II-1, II-2, II-3, or II-4, X ２ In each chemical formula, independently, N or CR ｘ And R ｘ is H, F or Cl, Br or I; R １ In each chemical formula, independently, -CX ａ H ２ , -C(X ａ ) ２ H, or -C(X ａ ) ３ Here, Xa is independently H, F, Cl, Br, or I in each chemical formula; R ２ and R ３ A compound represented by chemical formula I according to claim 1 or 3, wherein in each chemical formula, independently is F, Cl, Br, or I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

5. In the aforementioned chemical formulas II-1, II-2, II-3, or II-4, X ２ In each chemical formula, independently, N or CR ｘ And here, R ｘ is -H or F; R １ In each chemical formula, independently, -C(X ａ ) ２ H, or -C(X ａ ) ３ Here, Xa is independently F or Cl in each chemical formula; R ２ and R ３ In each of the equations, independently, F or Cl, A compound represented by chemical formula I as described in claim 4, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

6. Compounds having the structure represented by the following chemical formula I, their stereoisomers, or pharmaceutically acceptable salts thereof: Table 1

7. The compound described above is a compound represented by the following chemical formula, the compound according to claim 6, its stereoisomer or a pharmaceutically acceptable salt thereof: Table 2

8. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

9. The pharmaceutical composition according to claim 8, wherein the pharmaceutical composition is for the prevention or treatment of histone deacetylase 6-mediated diseases.

10. The aforementioned histone deacetylase 6-mediated diseases are Infectious diseases; neoplasms; endocrine, nutritional and metabolic disorders; mental and behavioral disorders; neurological disorders; ocular and adnexal disorders; cardiovascular diseases; respiratory diseases; digestive diseases; skin and subcutaneous tissue disorders; musculoskeletal and connective tissue disorders; or congenital malformations, deformities and infections. The pharmaceutical composition according to claim 9, which is a color abnormality.

11. The aforementioned infectious disease is a prion disease. The aforementioned neoplasm may be a benign or malignant tumor. The aforementioned endocrine, nutritional, and metabolic disorders include Wilson's disease, amyloidosis, or diabetes. The aforementioned mental and behavioral disorders are depression or Rett syndrome. The aforementioned neurological disorders include systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases, motor disorders, neuropathy, motor nerve disorders, or demyelinating diseases of the central nervous system. The aforementioned eye and adnexal disease is uveitis. The aforementioned cardiovascular disease is atrial fibrillation or stroke. The aforementioned respiratory disease is asthma. The aforementioned digestive diseases are alcoholic liver disease, inflammatory bowel disease, Crohn's disease, or ulcerative bowel disease. The aforementioned skin and subcutaneous tissue disease is psoriasis. The aforementioned musculoskeletal and connective tissue diseases are rheumatoid arthritis, osteoarthritis, or systemic lupus erythematosus. The aforementioned congenital malformations, deformities, and chromosomal abnormalities are autosomal dominant polycystic tumors. The pharmaceutical composition according to claim 10.

12. The aforementioned systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, is Huntington's disease, spinal muscular atrophy (SMA), or spinocerebellar ataxia (SCA). The aforementioned neurodegenerative disease is Alzheimer's disease or tauopathy. The aforementioned movement disorder is Parkinson's disease. The aforementioned neuropathic disorders include hereditary neuropathy, including Charcot-Marie-Tooth disease or hereditary spastic paraplegia, diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy, or drug-induced neuropathy. The aforementioned motor nerve disorder is amyotrophic lateral sclerosis (ALS). The aforementioned demyelinating disease of the central nervous system is multiple sclerosis (MS). The pharmaceutical composition according to claim 11.

13. A pharmaceutical composition for the prevention or treatment of systemic atrophy of the nervous system, including systemic atrophy of the central nervous system, neurodegenerative diseases, or neuropathy, comprising as an active ingredient a compound according to any one of claims 1 to 7, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

14. The aforementioned systemic atrophy of the central nervous system is Huntington's disease, spinal muscular atrophy (SMA), or spinocerebellar ataxia (SCA). The aforementioned neurodegenerative diseases include dementia, Alzheimer's disease, or tauopathy. The aforementioned neuropathy is hereditary neuropathy including Charcot-Marie-Tooth disease or hereditary spastic paraplegia, diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy, drug-induced neuropathy, amyotrophic lateral sclerosis (ALS), or multiple sclerosis (MS). The pharmaceutical composition according to claim 13.

15. A method for preventing or treating a histone deacetylase 6-mediated disease, comprising administering a therapeutically effective amount of a compound according to any one of claims 1 to 7, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

16. Use of a compound according to any one of claims 1 to 7, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, for the prevention or treatment of histone deacetylase 6-mediated diseases.

17. Use of a compound according to any one of claims 1 to 7, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, for the manufacture of an agent for the prevention or treatment of histone deacetylase 6-mediated diseases.

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