Oxadiazole derivative compound and pharmaceutical composition containing same

Abstract

The present invention relates to a novel compound having histone deacetylase 6 (HDAC6) inhibitory activity, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, use thereof for preparing a medicament, a pharmaceutical composition comprising the same, a prophylactic or therapeutic method thereof, and a method for preparing a novel 1, 3, 4-oxadiazole oxetane derivative.

Description

TECHNICAL FIELD

[0001] The present application relates to a novel oxadiazole derivative compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the oxadiazole derivative compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, a use of the oxadiazole derivative compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof for the preparation of a therapeutic drug, a method of preventing or treating a disease by administering the oxadiazole derivative compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a method for preparing the oxadiazole derivative compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. BACKGROUND

[0002] In cells, post-translational modifications, such as acetylation, play an extremely important regulatory module in the center of biological processes, and are also strictly controlled by various enzymes. As a core protein constituting chromatin, histone acts as an axle around which DNA is wound, and thus contributes to DNA condensation. In addition, the balance between acetylation and deacetylation of histone plays an extremely important role in gene expression.

[0003] As an enzyme that removes acetyl groups from lysine residues of histone proteins constituting chromatin, histone deacetylase (HDAC) is known to be associated with gene silencing and induces cell cycle arrest, angiogenesis inhibition, immunomodulation, apoptosis, etc. (Hassig et al., Curr. Opin. Chem. Biol. 1997, 1, 300-308). In addition, it is reported that inhibition of HDAC enzyme function induces cancer cell apoptosis by reducing the activity of cancer cell survival-related factors and activating in vivo cancer cell death-related factors (Warrell et al., J. Natl. Cancer Inst. 1998, 90, 1621-1625).

[0004] In the case of humans, 18 HDACs are known and are classified into four classes according to homology with yeast HDACs. In this case, eleven HDACs using zinc as a co-factor can be classified into three classes: Class I (HDAC1, 2, 3, 8), Class II (IIa: HDAC4, 5, 7, 9; IIb: HDAC6, 10), and Class IV (HDAC11). In addition, seven HDACs of Class III (SIRT 1 to 7) use NAD+ instead of zinc as a co-factor (Bolden et al., Nat. Rev. Drug Discov. 2006, 5(9), 769-784).

[0005] A variety of HDAC inhibitors are currently in preclinical or clinical development, but only non-selective HDAC inhibitors are known as anticancer agents to date. Vorinostat (SAHA) and romidepsin (FK228) have been approved as therapeutic agents for cutaneous T-cell lymphoma, and panobinostat (LBH-589) has been approved as a therapeutic agent for multiple myeloma. However, non-selective HDAC inhibitors are known to generally cause side effects such as fatigue, nausea, and the like at high doses (Piekarz et al., Pharmaceuticals 2010, 3, 2751-2767). It is reported that the side effects are caused by inhibition of class I HDACs. Due to such side effects, non-selective HDAC inhibitors are limited in drug development in other fields than anticancer agents (Witt et al., Cancer Letters 277 (2009) 8-21).

[0006] Meanwhile, it is reported that selective inhibition of class II HDACs does not show toxicity, whereas toxicity has occurred in the inhibition of class I HDACs. In the case of developing selective HDAC inhibitors, it is possible to solve side effects such as toxicity and the like caused by non-selective inhibition of HDACs. Therefore, there is an opportunity to develop selective HDAC inhibitors as effective therapeutic agents for various diseases (Matthias et al., Mol. Cell. Biol. 2008, 28, 1688-1701).

[0007] HDAC6 (one of class IIb HDACs) is known to exist mainly in the cytoplasm and to contain tubulin proteins, and thus to be involved in deacetylation of various non-histone substrates (HSP90, cortactin, and the like) (Yao et al., Mol. Cell 2005, 18, 601-607). HDAC6 has two catalytic domains, in which the C-terminal zinc finger domain can bind to ubiquitinated proteins. HDAC6 is known to have various non-histone proteins as substrates, and thus to play an important role in various diseases such as cancer, inflammatory diseases, autoimmune diseases, neurological diseases, neurodegenerative diseases, and the like (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 structural features common to various HDAC inhibitors include a cap group, a linker group, and a zinc-binding group (ZBG), as shown in the following structure of vorinostat. Many researchers have studied the inhibitory activity and selectivity of the enzyme through structural modification of the cap group and the linker group. In addition to these groups, the zinc-binding group is known to play a more important role in the 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]

[0010] Most of the zinc-binding groups include hydroxamic acid or benzamide, and in addition to this, hydroxamic acid derivatives show strong HDAC inhibitory effect, but have problems of low bioavailability and serious off-target activity. Benzamide contains aniline, and thus has a problem that a toxic metabolite can be produced in vivo (Woster et al., Med. Chem. Commun. 2015, online publication).

[0011] Therefore, unlike non-selective inhibitors having side effects, there is a need to develop selective HDAC6 inhibitors having a zinc-binding group with improved bioavailability, while not causing side effects, in order to treat cancer, inflammatory diseases, autoimmune diseases, neurological diseases, neurodegenerative diseases, etc.

[0012] [Related Art References]

[0013] [Patent Documents]

[0014] (Patent Document 1) International Unexamined Patent Publication No. WO 2011 / 091213

[0015] (Patent Document 2) International Unexamined Patent Publication No. WO 2011 / 011186

[0016] (Patent Document 3) International Unexamined Patent Publication No. WO 2013 / 052110

[0017] (Patent Document 4) International Unexamined Patent Publication No. WO 2013 / 041407

[0018] (Patent Document 5) International Unexamined Patent Publication No. WO 2013 / 134467

[0019] (Patent Document 6) International Unexamined Patent Publication No. WO 2013 / 008162

[0020] (Patent Document 7) International Unexamined Patent Publication No. WO 2013 / 080120

[0021] (Patent Document 8) International Unexamined Patent Publication No. WO 2013 / 066835

[0022] (Patent Document 9) International Unexamined Patent Publication No. WO 2013 / 066838

[0023] (Patent Document 10) International Unexamined Patent Publication No. WO 2013 / 066833

[0024] (Patent Document 11) International Unexamined Patent Publication No. WO 2013 / 066839 SUMMARY

[0025] TECHNICAL PROBLEM

[0026] An object of the present application is to provide an oxadiazole derivative compound having selective HDAC6 inhibitory activity, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0027] Another object of the present application 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.

[0028] Another object of the present application is to provide a method for preparing an oxadiazole derivative compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0029] Another object of the present application is to provide a pharmaceutical composition for preventing or treating a disease associated with HDAC6 activity or a degenerative brain disease, comprising an oxadiazole derivative compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0030] Another object of the present application is to provide use of an oxadiazole derivative compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for preventing or treating a disease associated with HDAC6 activity or a degenerative brain disease.

[0031] Another object of the present application is to provide a method for preventing or treating a disease associated with HDAC6 activity or a degenerative brain disease, comprising administering a therapeutically effective amount of an oxadiazole derivative compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0032] Another object of the present application is to provide use of an oxadiazole derivative compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, for preventing or treating a disease associated with HDAC6 activity or a degenerative brain disease.

[0033] TECHNICAL SOLUTION

[0034] The present inventors have discovered oxadiazole derivative compounds having histone deacetylase 6 (HDAC6) inhibitory activity, and have used them to inhibit or treat diseases associated with HDAC6 activity, thereby completing the present invention.

[0035] The present invention will be described in greater detail below. All combinations of the various elements disclosed in the present invention are within the scope of the present invention. In addition, it will be found that the scope of the present invention is not limited to the following detailed description.

[0036] Compounds

[0037] The present invention can provide any one of the following (1) to (7) compounds, stereoisomers thereof, or pharmaceutically acceptable salts thereof.

[0038] (1) a compound represented by the following formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0039] [Formula I]

[0040]

[0041] In the above formula I,

[0042] X1to X4are each independently N or CR x wherein three or more of X1to X4cannot be simultaneously N, and R x is -H, F, Cl, Br, or I;

[0043] R1is -CX a H2, -C(X a )2H, or -C(X a )3, wherein Xais F, Cl, Br, or I; and

[0044] R2and R3are each independently F, Cl, Br, or I.

[0045] (2) a compound represented by formula I according to the above (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0046] In the above formula I,

[0047] X1to X4are each independently N or -CR x wherein three or more of X1to X4cannot be simultaneously N, and R x is H or F;

[0048] R1is -CX a H2, or -C(X a )2H, wherein X a is F or Cl; and

[0049] R2and R3may each independently be F or Cl.

[0050] (3) The compound represented by formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to (1) or (2) above:

[0051] The compound represented by the above formula I can be a compound represented by formula II:

[0052] [Formula II]

[0053]

[0054] In the above formula II,

[0055] X2is N or CR x , wherein R x is H, F, Cl, Br, or I;

[0056] R1is -CX a H2, -C(X a )2H, or -C(X a )3, wherein Xais F, Cl, Br, or I; and

[0057] R2and R3may each independently be F, Cl, Br, or I.

[0058] (4) The compound represented by formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to (1), (2), or (3) above:

[0059] The compound represented by the above formula I or II can be a compound represented by the following formula II-1, II-2, II-3, or II-4.

[0060] [Formula II-1]

[0061]

[0062] [Formula II-2]

[0063]

[0064] [Formula II-3]

[0065]

[0066] [Formula II-4]

[0067]

[0068] In the above formula II-1, II-2, II-3, or II-4,

[0069] X2is independently N or CRx , and R x is H, F or Cl, Br or I;

[0070] R1in each formula is independently -CX a H2, -C(X a )2H or -C(X a )3, wherein Xain each formula is independently F, Cl, Br or I; and

[0071] R2and R3in each formula are each independently F, Cl, Br or I.

[0072] (5) The compound represented by formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to (1), (2), (3) or (4) described above:

[0073] In the above formula II-1, II-2, II-3 or II-4,

[0074] X2in each formula is independently N or -CR x , wherein R x is H or F;

[0075] R1in each formula is independently -CX a )2H or -C(X a )3, wherein X a is F or Cl; and

[0076] R2and R3in each formula are each independently F or Cl.

[0077] In an embodiment of the present application, in the above formula II-1, II-2, II-3 or II-4, R2and R3may be the same as or different from each other.

[0078] In an embodiment of the present application, in the above formula I, II or I-1, R2and R3may be the same as or different from each other, and particularly, R2and R3may be different from each other. For example, if one of R2and R3is F, the other can be Cl.

[0079] In an embodiment of the present application, in the above formula I, II, II-2, II-3 or II-4, R2and R3may be the same as or different from each other, and particularly, R2and R3may be the same as each other. For example, both of R2and R3may be F, or can be Cl.

[0080] (6) The compound represented by formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to (1), (2), (3), (4) or (5) described above:

[0081] wherein the compound represented by formula I of the present application can be any one selected from the group consisting of Compounds 1 to 8 shown in Table 1 below.

[0082] Table 1

[0083]

[0084]

[0085] (7) The compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the above (1), (2), (3), (4), (5), or (6):

[0086] wherein the compound represented by Formula I of the present application can be Compound 1 or 5 shown in Table 1 above.

[0087] In the present application, the term "pharmaceutically acceptable" can mean that it is physiologically acceptable and does not normally cause allergic reactions (e.g., gastrointestinal disorders and dizziness) or other similar reactions when administered into an individual.

[0088] The pharmaceutically acceptable salt of the present application can be prepared by a conventional method known to those skilled in the art.

[0089] In the present application, the pharmaceutically acceptable salt can refer to a salt conventionally used in the pharmaceutical industry, such as inorganic ion salts prepared from calcium, potassium, sodium, magnesium, etc.; inorganic acid salts prepared from hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, perchloric acid, sulfuric acid, etc.; organic acid salts prepared from 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, carbonic acid, vanillic acid, hydroiodic acid, etc.; sulfonic acid salts prepared from methane sulfonic acid, ethane sulfonic acid, benzene sulfonic acid, p-toluene sulfonic acid, naphthalene sulfonic acid, etc.; amino acid salts prepared from glycine, arginine, lysine, etc.; amine salts prepared from trimethylamine, triethylamine, ammonia, pyridine, methylpyridine, etc.; and the like, but the type of the salt referred to in the present application is not limited to those listed.

[0090] In the present application, the preferred salt can include hydrochloric acid, trifluoroacetic acid, citric acid, bromic acid, maleic acid, phosphoric acid, sulfuric acid, tartaric acid, etc.

[0091] The compound represented by Formula I, II, II-1, II-2, II-3, or II-4 of the present application can include at least one asymmetric carbon, and thus can exist as a racemate thereof, a racemic mixture, a single enantiomer (optical isomer), a mixture of diastereomers, and each diastereomer.

[0092] Such isomers can be separated by conventional techniques, for example, the compound represented by formula I, II, II-1, II-2, II-3, or II-4 can be separated by column chromatography, separation by HPLC, and the like. Alternatively, each stereoisomer of the compound represented by formula I, II, II-1, II-2, II-3, or II-4 can be stereospecifically synthesized using known arrangements of optically pure starting materials and / or reagents.

[0093] In the present application, "stereoisomer" can include diastereoisomer and optical isomer (enantiomer), wherein the optical isomer can include not only enantiomer but also a mixture of enantiomers, and even a racemate.

[0094] The compound represented by formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 listed in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof according to the present application can show histone deacetylase 6 (HDAC6) inhibitory activity, particularly selective inhibitory activity against HDAC6, and more particularly extremely high selective inhibitory activity against HDAC6 compared to other HDAC isoforms, and extremely high inhibitory activity against HDAC6 but little or no inhibitory activity against other HDAC isoforms (Tables 3, 4, and 5).

[0095] The compound represented by formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof according to the present application can significantly increase acetylation of tubulin in neural cells Figure 4 and Figure 5 ).

[0096] The compound represented by formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof according to the present application can show activity for preventing or treating a disease related to or mediated by histone deacetylase 6 (HDAC6) activity.

[0097] In the present application, "prevention" can mean all actions of inhibiting or delaying the occurrence of a disease by administering the compound represented by formula I, II, II-1, II-2, II-3, or II-4 according to the present application, the compounds 1 to 8 described in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof.

[0098] In the present application, "treatment" can mean all actions in which the suspected symptoms of the individual who can develop a disease or the symptoms of the individual who has a disease are improved or appear to be improved by administering the compound represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the present application.

[0099] In the present specification, "histone deacetylase-mediated disease" can mean a disease associated with histone deacetylase 6 (HDAC6) activity.

[0100] In the present application embodiment, the histone deacetylase 6 (HDAC6) activity-related disease or the histone deacetylase 6-mediated disease can include an infectious disease, a neoplasm, an endocrine disease, a nutritional and metabolic disease, a mental and behavioral disorder, a neurological disease, an eye and adnexa disease, a circulatory system disease, a respiratory system disease, a digestive problem, a skin and subcutaneous tissue disease, a musculoskeletal and connective tissue disease, or a teratosis, deformity, and chromosomal aberration, etc.

[0101] In the present application embodiment, the infectious disease can be a prion disease; the neoplasm can be a benign tumor or a malignant tumor; the endocrine disease, nutritional and metabolic disease can be Wilson's disease, amyloidosis, or diabetes; the mental and behavioral disorder can be depression or Rett syndrome; the neurological disease can be a neurological atrophy including central nervous system atrophy, a neurodegenerative disease, a movement disorder, a neuropathy, a motor neuron disease, or a central nervous system demyelinating disease; the eye and adnexa disease can be uveitis; the circulatory system disease can be atrial fibrillation or stroke; the respiratory system disease can be asthma; the digestive problem can be alcoholic liver disease, inflammatory bowel disease, Crohn's disease, or ulcerative bowel disease; the skin and subcutaneous tissue disease can be psoriasis; the musculoskeletal and connective tissue disease can be rheumatoid arthritis, osteoarthritis, or systemic lupus erythematosus; and the teratosis, deformity, and chromosomal aberration can be autosomal dominant polycystic kidney disease.

[0102] In embodiments of the present application, the neurodegeneration including central nervous system atrophy can be Huntington's disease, spinal muscular atrophy (SMA), or spinocerebellar ataxia (SCA); the neurodegenerative disease can be Alzheimer's disease or a tauopathy; the movement disorder can be Parkinson's disease; the neuropathic disease can be a hereditary neuropathy including Charcot-Marie-Tooth disease (peripheral neuropathy Charcot-Marie-Tooth disease, central neuropathy Charcot-Marie-Tooth disease) or hereditary spastic paraplegia, diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy, or drug-induced neuropathy; the motor neuropathy can be amyotrophic lateral sclerosis (ALS); and the central nervous system demyelinating disease can be multiple sclerosis (MS).

[0103] In embodiments of the present application, the disease related to the activity of histone deacetylase 6 (HDAC6) or the disease mediated by histone deacetylase 6, in addition to the symptoms or diseases associated with abnormal function of histone deacetylase 6, can include cancer, inflammatory disease, autoimmune disease, neurological or degenerative neurological disease, particularly lung cancer, colon cancer, breast cancer, prostate cancer, liver cancer, brain cancer, ovarian cancer, stomach cancer, skin cancer, pancreatic cancer, glioma, glioblastoma, leukemia, lymphoma, multiple myeloma, solid cancer, Wilson's disease, spinocerebellar ataxia, prion disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, hereditary neuropathy including Charcot-Marie-Tooth disease (peripheral neuropathy Charcot-Marie-Tooth disease, central neuropathy 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.

[0104] The compound represented by Formula I, II, II-1, II-2, II-3, or II-4, Compound 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the present application can show high brain-blood barrier permeability. In particular, the compound represented by Formula I, II, II-1, II-2, II-3, or II-4, Compound 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the present application can show a high B / P ratio after oral administration (Table 10).

[0105] The compound represented by formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the present application can show an excellent preventive and therapeutic effect on nervous system atrophy including central nervous system atrophy, neurodegenerative disease including degenerative brain disease, hereditary neuropathy, and the like.

[0106] In an embodiment of the present application, the nervous system atrophy including central nervous system atrophy, neurodegenerative disease, or hereditary neuropathy can be Huntington's disease, spinal muscular atrophy (SMA), spinocerebellar ataxia (SCA), Alzheimer's disease, tauopathy, hereditary neuropathy (including Charcot-Marie-Tooth disease (peripheral neuropathy Charcot-Marie-Tooth disease, central neuropathy Charcot-Marie-Tooth disease)), hereditary spastic paraplegia, diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy, drug-induced neuropathy, amyotrophic lateral sclerosis (ALS), or multiple sclerosis (MS).

[0107] In an embodiment of the present application, the compound represented by formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can show a remarkably excellent preventive and therapeutic effect on nervous system atrophy including central nervous system atrophy, neurodegenerative disease including degenerative brain disease, or hereditary neuropathy selected from the group consisting of Huntington's disease, dementia, Alzheimer's disease, amyloidosis, Charcot-Marie-Tooth disease (peripheral neuropathy Charcot-Marie-Tooth disease, central neuropathy Charcot-Marie-Tooth disease), and tauopathy.

[0108] In an embodiment of the present application, the compound represented by formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can increase the relative velocity of mitochondrial axons (the relative velocity of mitochondrial axons is reduced by treatment with a beta amyloid fragment (Aβ)), and this pharmacological effect can be sustained for a long period of time. Accordingly, the compound of the present application can exhibit an excellent preventive and therapeutic effect on nervous system atrophy (e.g., central nervous system atrophy including dementia and Alzheimer's disease), neurodegenerative disease, or hereditary neuropathy (Table 6, Table 7, Table 8, Figure 1 and Figure 2 ).

[0109] In embodiments of the present application, the compounds represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof can increase the reduced relative velocity of mitochondria axons in the nerve cells overexpressing tau protein. Thus, the compounds of the present application can exhibit excellent preventive and therapeutic effects on neuroatrophy (e.g., central nervous system atrophy, including tauopathy), neurodegenerative diseases, or genetic neuropathy (Table 9 and Figure 3 ).

[0110] In embodiments of the present application, the compounds represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof can significantly improve the decreased cognitive ability in tauopathy mice (PS19 mice). Thus, the compounds of the present application can exhibit excellent preventive and therapeutic effects on neuroatrophy (e.g., central nervous system atrophy, including tauopathy), neurodegenerative diseases, or genetic neuropathy ( Figure 6 、 Figure 7 and Figure 8 ).

[0111] In embodiments of the present application, the compounds represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof can significantly reduce the overphosphorylation of tau observed in the brain of tauopathy mice (PS19 mice). Thus, the compounds of the present application can exhibit excellent preventive and therapeutic effects on neuroatrophy (e.g., central nervous system atrophy, including tauopathy), neurodegenerative diseases including degenerative brain diseases, or genetic neuropathy ( Figure 9 、 Figure 10 、 Figure 11 、 Figure 12 and Figure 13 ).

[0112] In embodiments of the present application, the compounds represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof can significantly enhance the long-term memory of the hippocampus reduced in tauopathy mice (PS19 mice). Thus, the compounds of the present application can exhibit excellent preventive and therapeutic effects on neuroatrophy (e.g., central nervous system atrophy, including tauopathy), neurodegenerative diseases, or genetic neuropathy ( Figure 14 and Figure 15 ).

[0113] In embodiments of the present application, the compounds represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof can significantly improve the symptoms of ataxia in Huntington's disease model mice (Yac128 mice), and can significantly increase the grip strength reduced in Huntington's disease model mice (Yac128 mice). Thus, the compounds of the present application can exhibit excellent preventive and therapeutic effects on neurodegeneration (e.g., central nervous system atrophy, including Huntington's disease), neurodegenerative diseases, or hereditary neuropathies Figure 16 , Figure 17 , Figure 18 and Figure 19 .

[0114] In embodiments of the present application, the compounds represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof can significantly increase the relative velocity of mitochondria in the dorsal root ganglion (DRG) of Charcot-Marie-Tooth disease (CMT) model mice (MFN2 mutant mice). Thus, the compounds of the present application can exhibit excellent preventive and therapeutic effects on neurodegeneration (e.g., central nervous system atrophy, including Charcot-Marie-Tooth disease), neurodegenerative diseases, or hereditary neuropathies (Table 11 and Figure 20 .

[0115] In embodiments of the present application, the compounds represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof can significantly improve the reduced fall latency (confirmed by the constant speed rotarod test) in a CX32 deletion mouse model of Charcot-Marie-Tooth disease (CMT), and can significantly reduce the increased slip count and transverse time of the rod (confirmed by the balance beam test) in a CX32 deletion mouse model of Charcot-Marie-Tooth disease (CMT). Thus, the compounds of the present application can exhibit excellent preventive and therapeutic effects on neurodegeneration (e.g., central nervous system atrophy, including Charcot-Marie-Tooth disease), neurodegenerative diseases, or hereditary neuropathies Figure 21 and Figure 22 .

[0116] In embodiments of the present application, the compound represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can significantly improve the reduced latency to fall (confirmed by the accelerating rotarod test) in a MFN2 mutant mouse model of Charcot-Marie-Tooth disease (CMT) and can significantly reduce the increased slip count (confirmed by the balance beam test) in a MFN2 mutant mouse model of Charcot-Marie-Tooth disease (CMT). Thus, the compounds of the present application can exhibit excellent preventive and therapeutic effects on neurodegeneration (e.g., central nervous system atrophy, including Charcot-Marie-Tooth disease), neurodegenerative diseases, or hereditary neuropathies Figure 23 and Figure 24 ).

[0117] In embodiments of the present application, the compound represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can significantly improve the reduced latency to fall (confirmed by the accelerating rotarod test) in a CMT2A mouse model of Charcot-Marie-Tooth disease (CMT) and can significantly reduce the increased slip count (confirmed by the balance beam test) in a CMT2A mouse model of Charcot-Marie-Tooth disease (CMT). Thus, the compounds of the present application can exhibit excellent preventive and therapeutic effects on neurodegeneration (e.g., central nervous system atrophy, including Charcot-Marie-Tooth disease), neurodegenerative diseases, or hereditary neuropathies Figure 25 and Figure 26 ).

[0118] In embodiments of the present application, the compound represented by Formula I, II, II-1, II-2, II-3, or II-4, the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can significantly improve the reduced latency to fall (confirmed by the accelerating rotarod test) in a CMT2A mouse model of Charcot-Marie-Tooth disease (CMT) and can significantly reduce the increased slip count (confirmed by the balance beam test) in a CMT2A mouse model of Charcot-Marie-Tooth disease (CMT). Thus, the compounds of the present application can exhibit excellent preventive and therapeutic effects on neurodegeneration (e.g., central nervous system atrophy, including Charcot-Marie-Tooth disease), neurodegenerative diseases, or hereditary neuropathies Figure 27 and Figure 28 ).

[0119] In embodiments of the present application, the compound represented by Formula I, II, II-1, II-2, II-3, or II-4, Compound 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can significantly improve the reduced drop latency (confirmed by the constant rod test) in a CMT1A mouse model of Charcot-Marie-Tooth disease (CMT) and can significantly reduce the increased slip count (confirmed by the balance beam test) in a CMT1A mouse model of Charcot-Marie-Tooth disease (CMT). Thus, the compound of the present application can exhibit an excellent preventive and therapeutic effect on neurodegeneration (e.g., central nervous system atrophy, including Charcot-Marie-Tooth disease), neurodegenerative diseases, or hereditary neuropathy Figure 29 and Figure 30 ).

[0120] In embodiments of the present application, the compound represented by Formula I, II, II-1, II-2, II-3, or II-4, Compound 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can significantly improve the reduced sensory neuron action potential (SNAP) amplitude and sensory neuron conduction velocity (SNCV) in a MFN2 mutant mouse model of Charcot-Marie-Tooth disease (CMT). Thus, the compound of the present application can exhibit an excellent preventive and therapeutic effect on neurodegeneration (e.g., central nervous system atrophy, including Charcot-Marie-Tooth disease), neurodegenerative diseases, or hereditary neuropathy Figure 31 ).

[0121] In embodiments of the present application, the compound represented by Formula I, II, II-1, II-2, II-3, or II-4, Compound 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can significantly increase the reduced axon size of sciatic nerve fibers in a CX32 deletion mouse model of Charcot-Marie-Tooth disease (CMT). Thus, the compound of the present application can exhibit an excellent preventive and therapeutic effect on neurodegeneration (e.g., central nervous system atrophy, including Charcot-Marie-Tooth disease), neurodegenerative diseases, or hereditary neuropathy Figure 32 ).

[0122] Methods for preparing compounds

[0123] The present application can provide a method for preparing an oxadiazole derivative compound represented by Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0124] Hereinafter, in the reaction formula, symbols identical to those of Formula I and not specifically described are the same as those defined in Formula I, and repeated description is omitted. In addition, in the reaction formula, PG can represent an amine protecting group, and can be, for example, tert-butyloxycarbonyl (Boc).

[0125] The preferred method for preparing the oxadiazole derivative compound represented by the above Formula I, stereoisomer thereof, or pharmaceutically acceptable salt thereof can be the same as shown in the following Reaction Scheme 1-1, 1-2 to 1-3, and even therein can include a preparation method modified to a degree apparent to one skilled in the art.

[0126] In an embodiment of the present application, the preferred method for preparing the oxadiazole derivative compound represented by the above Formula I, stereoisomer thereof, or pharmaceutically acceptable salt thereof can be carried out by the preparation method of the following Reaction Scheme 1-1.

[0127] [Reaction Scheme 1-1]

[0128]

[0129] In the above [Reaction Scheme 1-1], X1 to X4, R1, R2, and R3 can be the same as those defined in Formula I, PG can be a protecting group, halogen can be F, Cl, Br, or I, and alkyl can be C1 to C5 alkyl.

[0130] In an embodiment of the present application, in the above [Reaction Scheme 1-1], X1, X3, and X4 can be CH, X2 can be N, R2 and R3 can each independently be F or Cl, R1 can be CF2H, PG can be tert-butyl carboxylate, and alkyl can be methyl, ethyl, or butyl.

[0131] In the above [Reaction Scheme 1-1], the compound represented by the above 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 can each independently be in the form of a salt, and the salt can be a hydrochloride or a trifluoroacetate.

[0132] The above [Reaction Scheme 1-1] can show a method for synthesizing a 1,3,4-oxadiazole derivative compound, in which a compound of Formula 1-1-1 containing an isocyanate can be reacted with a compound of Formula 1-1-2 in which a protecting group is introduced to prepare a compound of Formula 1-1-3 containing a urea structure.

[0133] Thereafter, the resulting compound can undergo a substitution reaction with a compound of Formula 1-1-4 to prepare a compound of Formula 1-1-5, and then the protecting group can be removed to prepare a compound of Formula 1-1-6.

[0134] The compound of formula 1-1-6 can undergo a reductive amination reaction with a compound of formula 1-1-7 to prepare a compound of formula 1-1-8, and then react with hydrazine to prepare a compound of formula 1-1-9, which is a hydrazine compound. Thereafter, difluoroacetic anhydride and imidazole can be used to prepare a compound of formula 1-1-10.

[0135] In an embodiment of the present application, in the reaction scheme [Reaction Scheme 1-1], the compound of formula 1-1-10 described above can be Compound 3.

[0136] In an embodiment of the present application, a preferred method for preparing an oxadiazole derivative compound represented by the above formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can be carried out by the preparation method of the following Reaction Scheme 1-2.

[0137] [Reaction Scheme 1-2]

[0138]

[0139] In the above [Reaction Scheme 1-2], X1 to X4, R1, R2, and R3 can be the same as those defined in formula I, PG can be a protecting group, halogen can be F, Cl, Br, or I, and alkyl can be C1 to C5 alkyl.

[0140] In an embodiment of the present application, in the above [Reaction Scheme 1-2], X1, X3, and X4 can be CH, X2 can be N or -CRx (Rx can be F or Cl, Br, or I), R2 and R3 can each independently be F or Cl, R1 can be CF2H, PG can be tert-butyl carboxylate, and alkyl can be methyl, ethyl, or butyl.

[0141] In the above [Reaction Scheme 1-2], the compound represented by 1-1-1, 1-2-1, 1-2-2, 1-1-4, 1-1-8, 1-1-9, or 1-1-10 described above can each independently be in the form of a salt, and the salt can be a hydrochloride or a trifluoroacetate.

[0142] The above [Reaction Scheme 1-2] can show a method for synthesizing a 1,3,4-oxadiazole compound having an oxetane structure, in which a compound of formula 1-1-1 containing an isocyanate can react with a compound of formula 1-2-1 having an oxetane introduced therein to prepare a compound of formula 1-2-2 containing a urea structure.

[0143] The compound of formula 1-2-2 can undergo a substitution reaction with a compound of formula 1-1-4 to prepare a compound 1-1-8, and then react with hydrazine to prepare a compound of formula 1-1-9, which is a hydrazine compound. Thereafter, difluoroacetic anhydride and imidazole can be used to prepare a target compound of formula 1-1-10.

[0144] In the present embodiment, in the above [Reaction Scheme 1-2], the compound of formula 1-1-10 can be compound 4, compound 5, compound 6, compound 7, compound 8, etc.

[0145] In the present embodiment, a preferred method for preparing the oxadiazole derivative compound represented by the above formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can be performed by the preparation method of the following Reaction Scheme 1-3.

[0146] [Reaction Scheme 1-3]

[0147]

[0148] In the above [Reaction Scheme 1-3], X1 to X4, R1, R2, and R3 can be the same as those defined in formula I, PG can be a protecting group, halogen can be F, Cl, Br, or I, and alkyl can be C1 to C5 alkyl.

[0149] In the present embodiment, in the above [Reaction Scheme 1-3], X1, X3, and X4 can be CH, X2 can be CH or -CRx(Rx can be F or Cl, Br, or I), R2 and R3 can each independently be F or Cl, R1 can be CF2H, PG can be tert-butyl carboxylate, and alkyl can be methyl, ethyl, or butyl.

[0150] In the above [Reaction Scheme 1-3], the compound represented by the above 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 can each independently be in the form of a salt, and the salt can be a hydrochloride or a trifluoroacetate.

[0151] The above [Reaction Scheme 1-3] can show a method for synthesizing an oxadiazole compound having an oxetane structure, in which the compound of formula 1-3-1 containing an amine group can undergo a substitution reaction with the compound of formula 1-1-4 to prepare the compound of formula 1-3-2, and then can react with the compound of formula 1-1-2 having a protecting group introduced therein to prepare the compound of formula 1-1-5 containing a urea structure. Thereafter, the protecting group of formula 1-1-5 can be removed to prepare the compound of formula 1-1-6.

[0152] The compound of formula 1-1-6 can undergo a reductive amination reaction with the compound of formula 1-1-7 to prepare the compound of formula 1-1-8, and then react with hydrazine to prepare the compound of formula 1-1-9, which is a hydrazine compound. Thereafter, difluoroacetic anhydride and imidazole can be used to prepare the target compound of formula 1-1-10.

[0153] In the present application, the compound of formula 1-1-10 can be Compound 1, Compound 2, and the like.

[0154] Pharmaceutical compositions, methods of treatment using the same and uses thereof

[0155] The present application can provide a pharmaceutical composition comprising a compound represented by the above formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0156] According to the present application, the compound represented by formula I contained in the pharmaceutical composition can be a compound represented by the above formula II.

[0157] According to the present application, the compound represented by formula I contained in the pharmaceutical composition can be a compound represented by the above formula II-1, II-2, II-3, or II-4.

[0158] The present application can provide a pharmaceutical composition comprising a compound of at least one of Compounds 1 to 8 described in Table 1 above, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0159] The present application can provide a pharmaceutical composition comprising a compound of at least one of Compounds 1 and 5 described in Table 1 above, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0160] The present application can provide a pharmaceutical composition for preventing or treating a histone deacetylase-mediated disease, comprising a compound represented by the above formula I, II, II-1, II-2, II-3, or II-4, a compound of at least one of Compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an effective ingredient.

[0161] The present application can provide a pharmaceutical composition for preventing or treating a histone deacetylase 6-mediated disease, comprising a compound represented by the above formula I, II, II-1, II-2, II-3, or II-4, a compound of at least one of Compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an effective ingredient.

[0162] The pharmaceutical composition of the present application can exhibit the same kind of pharmacological effects as those exhibited by a compound represented by the above formula I, II, II-1, II-2, II-3, or II-4, a compound of at least one of Compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0163] In the pharmaceutical composition, the histone deacetylase 6-mediated disease can be substantially the same as the histone deacetylase 6-mediated disease previously investigated in the compound.

[0164] In embodiments of the present application, the disease associated with histone deacetylase 6 (HDAC6) activity or the disease mediated by histone deacetylase 6 (HDAC6) can include infectious diseases, neoplasms, endocrine diseases, nutritional and metabolic diseases, mental and behavioral disorders, neurological diseases, diseases of the eye and adnexa, circulatory diseases, respiratory diseases, digestive problems, diseases of the skin and subcutaneous tissue, diseases of the musculoskeletal system and connective tissue, or malformations, deformations and chromosomal abnormalities, etc.

[0165] In embodiments of the present application, the infectious disease can be prion disease; the neoplasm can be benign tumor or malignant tumor; the endocrine disease, nutritional and metabolic disease can be Wilson disease, amyloidosis or diabetes; the mental and behavioral disorder can be depression or Rett syndrome; the neurological disease can be neurodegeneration including central nervous system atrophy, neurodegenerative disease, movement disorder, neuropathy, motor neuron disease or central nervous system demyelinating disease; the disease of the eye and adnexa can be uveitis; the circulatory disease can be atrial fibrillation or stroke; the respiratory disease can be asthma; the digestive problem can be alcoholic liver disease, inflammatory bowel disease, Crohn's disease or ulcerative colitis; the disease of the skin and subcutaneous tissue can be psoriasis; the disease of the musculoskeletal system and connective tissue can be rheumatoid arthritis, osteoarthritis or systemic lupus erythematosus; and the malformation, deformation and chromosomal abnormality can be autosomal dominant polycystic kidney disease.

[0166] In embodiments of the present application, the neurodegeneration including central nervous system atrophy can be Huntington's disease, spinal muscular atrophy (SMA) or spinocerebellar ataxia (SCA); the neurodegenerative disease can be Alzheimer's disease or tauopathy; the movement disorder can be Parkinson's disease; the neuropathy disease can be hereditary neuropathy including Charcot-Marie-Tooth disease (peripheral neuropathy Charcot-Marie-Tooth disease, central neuropathy Charcot-Marie-Tooth disease) or hereditary spastic paraplegia, diabetic neuropathy, sporadic neuropathy, inflammatory neuropathy or drug-induced neuropathy; the motor neuropathy can be amyotrophic lateral sclerosis (ALS); and the central nervous system demyelinating disease can be multiple sclerosis (MS).

[0167] In the embodiments of the present application, the disease associated with the histone deacetylase 6 (HDAC6) activity or the disease mediated by the histone deacetylase 6 (HDAC6) can include cancer, an inflammatory disease, an autoimmune disease, a neurological or degenerative neurological disease, specifically, lung cancer, colon cancer, breast cancer, prostate cancer, liver cancer, brain cancer, ovarian cancer, stomach cancer, skin cancer, pancreatic cancer, glioma, glioblastoma, leukemia, lymphoma, multiple myeloma, solid cancer, Wilson's disease, spinocerebellar ataxia, prion disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, hereditary neuropathy (including Charcot-Marie-Tooth disease (peripheral neuropathy Charcot-Marie-Tooth disease, central neuropathy Charcot-Marie-Tooth disease)) or hereditary spastic paraplegia, diabetic neuropathy, idiopathic neuropathy, inflammatory neuropathy, or drug-induced neuropathy, amyloidosis, Alzheimer's disease, alcoholic liver disease, spinal muscular atrophy, rheumatoid arthritis or osteoarthritis, and a symptom or disease associated with abnormal function of the histone deacetylase 6.

[0168] The present application can provide a pharmaceutical composition for preventing or treating a neurological atrophy including central nervous system atrophy, a neurodegenerative disease including a degenerative brain disease, or a hereditary neuropathy, which comprises a compound represented by the above-described formula I, II, II-1, II-2, II-3, or II-4, a compound of at least one of compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an effective ingredient.

[0169] In the pharmaceutical composition, the neurological atrophy, the neurodegenerative disease, and the hereditary neuropathy can be substantially the same as the degenerative brain disease previously studied in the compound.

[0170] In the embodiments of the present application, the degenerative brain disease can be Huntington's disease, spinal muscular atrophy (SMA), spinocerebellar ataxia (SCA), Alzheimer's disease, tauopathy, hereditary neuropathy (including Charcot-Marie-Tooth disease (peripheral neuropathy Charcot-Marie-Tooth disease, central neuropathy Charcot-Marie-Tooth disease)) or hereditary spastic paraplegia, diabetic neuropathy, idiopathic neuropathy, inflammatory neuropathy, drug-induced neuropathy, amyotrophic lateral sclerosis (ALS), or multiple sclerosis (MS), and particularly, Huntington's disease, dementia, Alzheimer's disease, amyloidosis, or tauopathy.

[0171] In the pharmaceutical composition, the stereoisomer and the pharmaceutically acceptable salt can be the same as described above in the stereoisomer and the pharmaceutically acceptable salt in the compound.

[0172] In addition to compounds represented by formulas I, II, II-1, II-2, II-3 or II-4 above, compounds of at least one of compounds 1 to 8 described in Table 1, their stereoisomers or pharmaceutically acceptable salts thereof, the pharmaceutical compositions of the present invention may further comprise at least one pharmaceutically acceptable additive.

[0173] In embodiments of the present invention, pharmaceutically acceptable additives may include saline solutions, sterile water, Ringer's solution, buffered saline, glucose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of these components, and may include other conventional additives, such as antioxidants, buffer solutions, and antibacterial agents, if necessary. Furthermore, diluents, dispersants, surfactants, binders, and lubricants may be added to formulate injectable formulations, such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, or tablets. Therefore, the compositions of the present invention can be patches, liquids, pills, capsules, granules, tablets, suppositories, etc. Formulations can be prepared according to conventional methods used in the art for formulation or methods disclosed in Remington's Pharmaceutical Science (latest version), Mack Publishing Company, Easton PA, and the compositions can be formulated into various formulations according to various diseases or ingredients.

[0174] In embodiments of the present invention, pharmaceutically acceptable additives that may be included in the pharmaceutical composition may be those conventionally used in the art, specifically including (but not limited to) lactose, glucose, sucrose, sorbitol, mannitol, glycine, starch, tragacanth rubber, acacia rubber, calcium phosphate, calcium chloride, sodium chloride, alginic acid, sodium alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, polyethylene glycol, cellulose, water, ethanol, syrup, methylcellulose, methylparaben, propylparaben, talc, magnesium stearate, magnesium aluminum silicate, silicon dioxide, orange flavor, strawberry flavor, vanilla flavor, or mineral oil.

[0175] The pharmaceutical compositions of the present invention may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or locally) according to the intended method, wherein the dosage may vary within its range based on factors (including patient weight, age, sex, health status, diet, time of administration, method of administration, route of administration, excretion rate, disease type, disease severity, duration of treatment, combination or concurrent use of drugs) and other factors well known in the medical field, and may be determined by a person skilled in the art considering all of the foregoing factors.

[0176] The daily dose of the pharmaceutical composition comprising the compound represented by the above-described formula I, II, II-1, II-2, II-3, or II-4, at least one of the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the present application can be about 1 mg / kg to 1000 mg / kg, preferably about 5 mg / kg to 100 mg / kg, and can be administered once a day or several times a day by dividing the daily dose of the composition.

[0177] In addition to the compound represented by the above-described formula I, II, II-1, II-2, II-3, or II-4, at least one of the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, the pharmaceutical composition of the present application can further comprise at least one effective ingredient showing the same or similar efficacy.

[0178] The present application can provide a method for preventing or treating a disease associated with HDAC6 activity or a nervous system atrophy including central nervous system atrophy, a neurodegenerative disease including a degenerative brain disease, or a hereditary neuropathy, which comprises administering to an individual a compound represented by the above-described formula I, II, II-1, II-2, II-3, or II-4, at least one of the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0179] The method for preventing or treating a disease associated with histone deacetylase 6 activity or a degenerative brain disease according to the present application can not only include treating the disease itself before the symptoms appear by administering a compound represented by the above-described formula I, II, II-1, II-2, II-3, or II-4, at least one of the compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, but also inhibits or prevents such symptoms. In the management of the disease, the prophylactic or therapeutic dose of the specific active ingredient can vary depending on the nature and severity of the disease or condition and the route of administration of the active ingredient. The dose and frequency thereof can vary depending on the age, weight, and response of the individual patient. The appropriate dose and usage can be easily selected by those skilled in the art with reasonable consideration of such factors.

[0180] In the method of the present application, the disease associated with histone deacetylase 6 activity or a degenerative brain disease can be the same as described above.

[0181] In the present application, "administration" can mean introducing a predetermined substance into an individual by an appropriate method.

[0182] In the present invention, "individual" can mean all animals, such as rats, mice, domestic animals, etc., including humans, which can form or have formed a disease related to HDAC6 activity or a degenerative brain disease, and particularly can mean mammals, including humans, but is not limited thereto.

[0183] The method for preventing or treating a disease related to HDAC6 activity of the present invention or a nervous system atrophy including central nervous system atrophy, a neurodegenerative disease including a degenerative brain disease, or a hereditary neuropathy can mean administration of a therapeutically effective amount of a compound represented by the above Formula I, II, II-1, II-2, II-3, or II-4, a compound of at least one of Compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0184] In the present invention, "therapeutically effective amount" can mean an amount sufficient to treat a disease at a reasonable risk / benefit ratio applicable to medical treatment without causing side effects, and can mean an amount of a compound represented by the above Formula I, II, II-1, II-2, II-3, or II-4, a compound of at least one of Compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, which is effective for preventing or treating a disease related to histone deacetylase 6 activity or a nervous system atrophy including central nervous system atrophy, a neurodegenerative disease including a degenerative brain disease, or a hereditary neuropathy.

[0185] Further, the method for preventing or treating a disease related to histone deacetylase 6 activity of the present invention or a nervous system atrophy including central nervous system atrophy, a neurodegenerative disease including a degenerative brain disease, or a hereditary neuropathy can further include administration of a therapeutically effective amount of an additional active agent that is helpful in treating the disease, as well as a compound represented by the above Formula I, II, II-1, II-2, II-3, or II-4, a compound of at least one of Compounds 1 to 8 described in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and the additional active agent can show a synergistic effect, an additive effect, or a supplementary effect together with the compound represented by the above Formula I, II, II-1, II-2, II-3, or II-4, the compound of at least one of Compounds 1 to 8 described in Table 1, the stereoisomer thereof, or the pharmaceutically acceptable salt thereof.

[0186] The present invention can provide the use of compounds represented by formulas I, II, II-1, II-2, II-3 or II-4 above, compounds of at least one of compounds 1 to 8 described in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising them for the prevention or treatment of HDAC6 activity-related diseases or nervous system atrophy including central nervous system atrophy, neurodegenerative diseases including degenerative brain diseases or hereditary neuropathies.

[0187] The present invention can provide the use of compounds represented by formulas I, II, II-1, II-2, II-3 or II-4 above, compounds of at least one of compounds 1 to 8 described in Table 1, their stereoisomers, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising them, for the preparation of medicaments for the prevention or treatment of HDAC6-related diseases or nervous system atrophy, including central nervous system atrophy, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathies.

[0188] In the application of this invention, histone deacetylase 6 activity-related diseases or nervous system atrophy including central nervous system atrophy, neurodegenerative diseases including degenerative brain diseases, or hereditary neuropathies can be the same as described above.

[0189] In order to prepare a drug, the compounds of the present invention represented by formulas I, II, II-1, II-2, II-3 or II-4 above, at least one of compounds 1 to 8 described in Table 1, their stereoisomers or pharmaceutically acceptable salts thereof can be mixed with pharmaceutically acceptable excipients, diluents, carriers, etc., and can be prepared together with other active agents to form a complex formulation, thereby providing a synergistic effect.

[0190] The same applies to any of the claims mentioned in the various aspects of this invention (i.e., oxadiazole derivative compounds, methods of their preparation, pharmaceutical compositions comprising them, treatment methods using them, and their uses) unless they contradict each other.

[0191] Beneficial effects

[0192] The novel oxadiazole derivative compounds, their stereoisomers, or pharmaceutically acceptable salts thereof according to the present invention may have selective HDAC6 inhibitory activity and show excellent preventive or therapeutic effects on HDAC6 activity-related diseases or nervous system atrophy, including central nervous system atrophy, neurodegenerative diseases, including degenerative brain diseases, or hereditary neuropathies. Attached Figure Description

[0193] Figure 1 and Figure 2Results showing the effect of compounds of the application on mitochondrial axonal relative velocity, which is reduced by treatment with Aβ in mouse hippocampal neurons, are shown.

[0194] Figure 3 Results showing the effect of compounds of the application on mitochondrial axonal relative velocity, which is reduced in mouse primary culture cells overexpressing human tau protein with the P301 L mutation, are shown.

[0195] Figure 4 and Figure 5 Results showing the effect of compounds of the application on acetylation of tubulin in SH-SY5Y cells (human neuroblastoma) are shown.

[0196] Figure 6 to Figure 8 Results showing the effect of compounds of the application on the decrease in cognitive ability due to tauopathy in PS19 mice are shown.

[0197] Figure 9 to Figure 13 Results showing the effect of compounds of the application on hyperphosphorylation of tau in the brain of PS19 mice are shown.

[0198] Figure 14 and Figure 15 Results showing the effect of compounds of the application on the decrease in hippocampal long-term memory enhancement in PS19 mice are shown.

[0199] Figure 16 to Figure 19 Results showing the effect of compounds of the application on the decrease in motor function in Yac128 mice are shown.

[0200] Figure 20 Results showing the effect of compounds of the application on mitochondrial axonal relative velocity, which is reduced in a model of Charcot-Marie-Tooth disease (MFN2 mutant mice), are shown.

[0201] Figure 21 and Figure 22 Results showing the effect of compounds of the application on the decrease in motor and sensory functions (rotarod, balance beam test) in a model of Charcot-Marie-Tooth disease (CX32 deficient mice) are shown.

[0202] Figure 23 and Figure 24 Results showing the effect of compounds of the application on the decrease in motor and sensory functions (rotarod, balance beam test) in a model of Charcot-Marie-Tooth disease (MFN2 mutant mice) are shown.

[0203] Figure 25 and Figure 26Results showing the evaluation of the effect of the compounds of the present application on reduced motor and sensory functions (rotarod, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CMT2A mice).

[0204] Figure 27 and Figure 28 Results showing the evaluation of the effect of the compounds of the present application on reduced motor and sensory functions (rotarod, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CMT1X mice).

[0205] Figure 29 and Figure 30 Results showing the evaluation of the effect of the compounds of the present application on reduced motor and sensory functions (rotarod, balance beam test) in an animal model of Charcot-Marie-Tooth disease (CMT1A mice).

[0206] Figure 31 Results showing the evaluation of the effect of the compounds of the present application on reduced nerve conduction velocity in an animal model of Charcot-Marie-Tooth disease (MFN2 mutant mice).

[0207] Figure 32 Results showing the evaluation of the effect of the compounds of the present application on reduced axonal size of sciatic nerve fibers in an animal model of Charcot-Marie-Tooth disease (CX32 deletion mice). DETAILED DESCRIPTION

[0208] Hereinafter, the present application is described in more detail by way of illustrative embodiments. These illustrative embodiments are provided for the purpose of illustration only, and thus, it will be apparent to those skilled in the art that the scope of the present application is not limited to the same.

[0209] Preparation of compounds

[0210] A specific method for preparing the compound represented by Formula I is as follows.

[0211] Example 1: Synthesis of Compound 1 (Compound 1), i.e., N-(3-chloro-4-fluorophenyl)-N-(4-(5-(difluoromethyl)-1,3,4-oxadiazol-2-yl)benzyl)-4-(oxetan-3-yl)piperazine-1-carboxamide

[0212] [Step 1] Synthesis of methyl 4-(((3-chloro-4-fluorophenyl)amino)methyl)benzoate

[0213]

[0214] 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, after which 4-(bromomethyl)benzoic acid methyl ester (4.581 g, 20.000 mmol) was added thereto and further stirred at the same temperature for 18 hours. Water was poured into the reaction mixture and extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting concentrate was purified by column chromatography (SiO2, 40 g column; ethyl acetate / hexane = 0 to 10%) and concentrated to obtain the title compound as a brown oil (3.711 g, 63.2%).

[0215] [Step 2] Synthesis of 4-((3-chloro-4-fluorophenyl)(4-(methoxycarbonyl)benzyl)carbamoyl)piperazine-1-carboxylic acid tert-butyl ester

[0216]

[0217] A solution of 4-(((3-chloro-4-fluorophenyl)amino)methyl)benzoic acid methyl ester (3.711 g, 12.635 mmol) prepared in Step 1, N,N-diisopropylethylamine (4.401 mL, 25.270 mmol), and triphosgene (1.875 g, 6.317 mmol) dissolved in dichloromethane (50 mL) was stirred at room temperature for 10 minutes, after which piperazine-1-carboxylic acid tert-butyl ester (2.353 g, 12.635 mmol) was added thereto and further stirred at room temperature for 18 hours. The solvent was removed from the reaction mixture under reduced pressure, and then the title compound was used without an additional purification process (6.300 g, 98.5%, brown oil).

[0218] [Step 3] Synthesis of 4-((N-(3-chloro-4-fluorophenyl)piperazine-1-carboxamido)methyl)benzoic acid methyl ester hydrochloride

[0219]

[0220] A solution of 4-((3-chloro-4-fluorophenyl)(4-(methoxycarbonyl)benzyl)carbamoyl)piperazine-1-carboxylic acid tert-butyl ester (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) dissolved in dichloromethane (50 mL) at room temperature was stirred at the same temperature for 3 hours. The precipitated solid was filtered, washed with dichloromethane, and dried to obtain the title compound as a white solid (3.647 g, 66.2%).

[0221] [Step 4] Synthesis of methyl 4-((N-(3-chloro-4-fluorophenyl)-4-(oxetan-3- yl)piperazine-1 -carboxamido)methyl)benzoate

[0222]

[0223] A solution of methyl 4-((N-(3-chloro-4-fluorophenyl)piperazine-1 - carboxamido)methyl)benzoate hydrochloride (0.885 g, 2.000 mmol) prepared in Step 3, oxetan-3-one (0.234 mL, 4.000 mmol) and sodium triacetoxyborohydride (0.848 g, 4.000 mmol) dissolved in dichloromethane (10 mL) at room temperature was stirred at the same temperature for 18 hours. Water was poured into the reaction mixture, extracted with dichloromethane and filtered through a plastic filter to remove solid residues and aqueous solution layer therefrom, and then concentrated under reduced pressure. The resulting concentrate was purified by column chromatography (Si02, 12 g cartridge; methanol / dichloromethane = 0 to 5%) and concentrated to obtain the title compound (0.626 g, 67.7%) as a brown oil.

[0224] [Step 5] Synthesis of N-(3-chloro-4-fluorophenyl)-N-(4-(hydrazinecarbonyl)benzyl)- 4-(oxetan-3-yl)piperazine-1 -carboxamide

[0225]

[0226] A solution of methyl 4-((N-(3-chloro-4-fluorophenyl)-4-(oxetan-3- yl)piperazine-1 -carboxamido)methyl)benzoate (0.626 g, 1.355 mmol) prepared in Step 4 and hydrazine monohydrate (1.317 mL, 27.104 mmol) dissolved in ethanol (6 mL) at room temperature was stirred at 75 °C for 18 hours, after which the reaction was terminated by reducing the temperature to room temperature. The solvent was removed from the resulting mixture under reduced pressure, and then the resulting concentrate was purified by column chromatography (Si02, 12 g cartridge; methanol / dichloromethane = 0 to 10%) and concentrated to obtain the title compound (0.435 g, 69.5%) as a white solid.

[0227] [Step 6] Synthesis of Compound 1

[0228]

[0229] A solution of N-(3-chloro-4-fluorophenyl)-N-(4-(hydrazinecarbonyl)benzyl)-4- (oxetan-3-yl)piperazine-1 -carboxamide prepared in Step 5 (0.100 g, 0.216 mmol), triethylamine (0.091 mL, 0.649 mmol), and 2,2-difluoroacetic anhydride (0.081 mL, 0.649 mmol) dissolved in dichloromethane (2 mL) at room temperature was stirred at 40 °C for 18 h, after which the reaction was completed by reducing the temperature to room temperature. Water was poured into the reaction mixture, extracted with dichloromethane, filtered through a plastic filter to remove solid residue and aqueous solution layers therefrom, and concentrated under reduced pressure. The resulting concentrate was purified by column chromatography (Si02, 4 g column; ethyl acetate / hexanes = 0 to 60%) and concentrated to obtain Compound 1 as a white solid (0.099 g, 87.9%).

[0230] 1 H NMR (400 MHz, CDC13) 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, 2H), 4.68 (t, 2H, J = 6.9 Hz), 3.68 - 3.65 (m, 1H), 3.49 - 3.47 (m, 4H), 2.43 (br s, 4H); LRMS (ES) m / z 522.4 (M + +1).

[0231] Example 2: Synthesis of Compound 2, N-(3-chloro-4-fluorophenyl)-N-(4-(5-(difluoromethyl)- 1,3,4-oxadiazol-2-yl)-2-fluorobenzyl)-4-(oxetan-3-yl)piperazine-1-carboxamide

[0232] Compound 2 was prepared according to the same reactions as described in Example 1, Steps 1 to 6, except that 4-(bromomethyl)-3-fluorobenzoic acid methyl ester was used in place of 4-(bromomethyl)benzoic acid methyl ester in Step 1 of Example 1.

[0233] 1H NMR (400 MHz, CDC13) 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, 2H), 4.67 (t, 2H, J = 6.9 Hz), 3.68 - 3.61 (m, 1H), 3.47-3.45 (m, 4H), 2.41 (br s, 4H); LRMS (ES) m / z 540.4 (M + +1).

[0234] Example 3: Synthesis of compound 3, i.e. N-(3-chloro-4-fluorophenyl)-N-((5-(5- (dichloromethyl)-1,3,4-oxadiazol-2-yl)pyridin-2-yl)methyl)-4-(oxetan-3-yl)piperazine-1- carboxamide

[0235] [Step 1] Synthesis of tert-butyl 4-((3-chloro-4-fluorophenyl)((5- (methoxycarbonyl)pyridin-2-yl)methyl)carbamoyl)piperazine-1-carboxylate

[0236]

[0237] Methyl 6-(bromomethyl)nicotinate (1.013 g, 4.402 mmol) was added to a solution of tert-butyl 4-((3-chloro-4-fluorophenyl)carbamoyl)piperazine-1-carboxylate (1.500 g, 4.192 mmol) and sodium hydride (60.00%, 0.184 g, 4.611 mmol) dissolved in N,N-dimethylformamide (30 mL) at 0 °C and stirred at the same temperature for 1 hour. Saturated aqueous sodium bicarbonate solution was poured into the reaction mixture and extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium chloride solution, anhydrous magnesium sulfate was added to remove water, filtered, and concentrated under reduced pressure. The resulting concentrate was purified by column chromatography (SiO2, 24 g column; ethyl acetate / hexane = 20% to 60%) and concentrated to obtain the title compound (2.000 g, 94.1%) as a yellow solid.

[0238] [Step 2] Synthesis of methyl 6-((N-(3-chloro-4-fluorophenyl)piperazine-1-carboxamido)methyl)nicotinate hydrochloride

[0239]

[0240] A solution of tert-butyl 4-((3-chloro-4-fluorophenyl)((5- (methoxycarbonyl)pyridin-2-yl)methyl)aminocarbonyl)piperazine-1-carboxylate prepared in Step 1 (2.000 g, 3.945 mmol) and hydrochloric acid (4.00 M solution in 1,4-dioxane, 4.931 mL, 19.725 mmol) dissolved in dichloromethane (50 mL) at room temperature was stirred at the same temperature for 18 hours. Saturated aqueous sodium bicarbonate solution was poured into the reaction mixture and extracted with dichloromethane. The organic layer was washed with saturated aqueous sodium chloride solution, anhydrous sodium sulfate was added to dryness, filtered and concentrated under reduced pressure. Ethyl acetate was put into the resulting concentrate and stirred to filter the precipitated solid which was then washed with ethyl acetate and subsequently dried to obtain the title compound as a yellow solid (1.120 g, 64.0%).

[0241] [Step 3] Synthesis of methyl 6-((N-(3-chloro-4-fluorophenyl)-4- (oxetan-3-yl)piperazine-1-carboxamido)methyl)nicotinate

[0242]

[0243] Oxetan-3-one (0.194 mL, 3.032 mmol) was added to a solution of methyl 6-((N-(3-chloro-4-fluorophenyl)piperazine-1-carboxamido)methyl)nicotinate hydrochloride prepared in Step 2 (1.120 g, 2.527 mmol) and N,N-diisopropylethylamine (0.440 mL, 2.527 mmol) dissolved in dichloromethane (20 mL) at room temperature and stirred at the same temperature. Sodium triacetoxyborohydride (0.803 g, 3.790 mmol) was added to the reaction mixture and further stirred at the same temperature for 18 hours. Saturated aqueous sodium bicarbonate solution was poured into the reaction mixture, extracted with dichloromethane, filtered through a plastic filter to remove solid residues and aqueous solution layers therefrom and concentrated under reduced pressure. The resulting concentrate was purified by column chromatography (SiO2, 24 g cartridge; ethyl acetate = 100%) and concentrated to obtain the product which was then purified again by chromatography (SiO2, 24 g cartridge; methanol / dichloromethane = 0 to 10%) and concentrated to obtain the title compound as a yellow oil (0.467 g, 39.9%).

[0244] [Step 4] Synthesis of N-(3-chloro-4-fluorophenyl)-N-((5-(hydrazinecarbonyl)pyridin-2- yl)methyl)-4-(oxetan-3-yl)piperazine-1-carboxamide

[0245]

[0246] A solution of methyl 6-((N-(3-chloro-4-fluorophenyl)-4-(oxetan-3-yl)piperazine-l- carboxamido)methyl)nicotinate (0.467 g, 1.009 mmol) and hydrazine monohydrate (0.981 mL, 20.177 mmol) prepared in Step 3 dissolved in ethanol (4 mL) at room temperature was stirred at 110 °C for 18 h, after which the reaction was ended by reducing the temperature to room temperature. The solvent was removed from the reaction mixture under reduced pressure, and then the title compound was used without further purification process (0.460 g, 98.5%, yellow solid).

[0247] [Step 5] Synthesis of Compound 3

[0248]

[0249] A solution of N-(3-chloro-4-fluorophenyl)-N-((5-(hydrazinecarbonyl)pyridin-2- yl)methyl)-4-(oxetan-3-yl)piperazine-1 -carboxamide (0.470 g, 1.015 mmol) and imidazole (0.207 g, 3.046 mmol) prepared in Step 4 dissolved in dichloromethane (10 mL) at room temperature was added 2,2-difluoroacetic anhydride (0.379 mL, 3.046 mmol) and refluxed (under heating) for 18 h, after which the reaction was ended by reducing the temperature to room temperature. Saturated aqueous sodium bicarbonate solution was poured into the reaction mixture, extracted with dichloromethane, filtered through a plastic filter to remove solid residues and aqueous solution layer therefrom, and concentrated under reduced pressure. The resulting concentrate was purified by column chromatography (Si02, 12 g column; methanol / dichloromethane = 0 to 2.5%) and concentrated to obtain Compound 3 as a yellow solid (0.167 g, 31.5%).

[0250] 1 H NMR (400 MHz, CDC13) 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).

[0251] Example 4: Synthesis of compound 4, i.e. N-(3,4-dichlorophenyl)-N-(4-(5- (difluoromethyl)-1,3,4-oxadiazol-2-yl)-2-fluorobenzyl)-4-(oxetan-3-yl)piperazine-1- carboxamide

[0252] [Step 1] Synthesis of tert-butyl 4-(oxetan-3-yl)piperazine-1-carboxylate

[0253]

[0254] Sodium triacetoxyborohydride (11.379 g, 53.688 mmol) was added to a solution of tert-butyl piperazine-1-carboxylate (5.000 g, 26.844 mmol) and oxetan-3-one (2.902 g, 40.266 mmol) dissolved in dichloromethane (200 mL) at room temperature and stirred at the same temperature for 18 hours. Saturated aqueous sodium bicarbonate solution was poured into the reaction mixture and extracted with dichloromethane. The organic layer was washed with saturated aqueous sodium chloride solution, anhydrous magnesium sulfate was added to remove water, filtered, and concentrated under reduced pressure. Ethyl ether was added to the resulting concentrate and stirred to filter the precipitated solid, which was then washed with ethyl ether and subsequently dried to obtain the title compound (6.230 g, 95.8%) in the form of a white solid.

[0255] [Step 2] Synthesis of 1-(oxetan-3-yl)piperazine trifluoroacetate

[0256]

[0257] 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. The solvent was removed from the reaction mixture under reduced pressure, and then ethyl acetate was added to the resulting concentrate and stirred to filter the precipitated solid, which was then washed with ethyl acetate and dried to obtain the title compound (4.720 g, 76.7%) in the form of a white solid.

[0258] [Step 3] Synthesis of N-(3,4-dichlorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide

[0259]

[0260] A solution of 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, dissolved in diethyl ether (10 mL) at room temperature, was stirred at the same temperature for 2 hours. The resulting precipitated solid was filtered, washed with diethyl ether, and dried to obtain the title compound as a white solid (0.497 g, 94.3%).

[0261] [Step 4] Synthesis of N-(3,4-dichlorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide

[0262]

[0263] Methyl 4-(bromomethyl)-3-fluorobenzoate (0.173 g, 0.700 mmol) was added to a solution of N-(3,4-dichlorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamide (0.210 g, 0.636 mmol) and sodium cyanide (60.00%, 0.028 g, 0.700 mmol) prepared in Step 3, dissolved in N,N-dimethylformamide (4 mL) at 0 °C and stirred at the same room temperature for 18 hours. The solvent was removed from the reaction mixture under reduced pressure, and then water was poured into the resulting concentrate, extracted with dichloromethane, filtered through a plastic filter to remove solid residues and aqueous solution layers therefrom, and concentrated under reduced pressure. The resulting concentrate was purified by column chromatography (SiO2, 4 g column; methanol / dichloromethane = 0 to 5%) and concentrated to obtain the title compound as a light yellow solid (0.170 g, 53.8%).

[0264] [Step 5] Synthesis of N-(3,4-dichlorophenyl)-N-(2-fluoro-4-(hydrazinecarbonyl)benzyl)-4- (oxetan-3-yl)piperazine-1-carboxamide

[0265]

[0266] A solution of 4-((N-(3,4-dichlorophenyl)-4-(oxetan-3-yl)piperazine-1-carboxamido)methyl)- 3-fluorobenzoic acid methyl ester (0.170 g, 0.342 mmol) prepared in Step 4 and hydrazine monohydrate (0.333 mL, 6.850 mmol), dissolved in ethanol (5 mL) at room temperature, was stirred at 110 °C for 18 hours, after which the reaction was completed by reducing the temperature to room temperature. The solvent was removed from the reaction mixture under reduced pressure, and then the title compound was used without an additional purification process (0.170 g, 100.0%, light yellow solid).

[0267] [Step 6] Synthesis of compound 4

[0268]

[0269] To a solution of N-(3,4-dichlorophenyl)-N-(2-fluoro-4-(hydrazinecarbonyl)benzyl)- 4-(oxetan-3-yl)piperazine-1 -carboxamide (0.170 g, 0.342 mmol) prepared in Step 5 and imidazole (0.070 g, 1.027 mmol) dissolved in dichloromethane (4 mL) at room temperature was added 2,2-difluoroacetic anhydride (0.128 mL, 1.027 mmol) and heated at reflux for 18 hours after which the reaction was ended by reducing the temperature to room temperature. The solvent was removed from the resulting mixture under reduced pressure and the resulting concentrate was then purified by column chromatography (Si02, 4 g column; methanol / dichloromethane = 0 to 2.5%) and concentrated to obtain the desired compound 4 (0.020 g, 10.5%) as a white solid.

[0270] 1 H NMR (400 MHz, CDC13) δ 7.89 (d, J = 7.8 Hz, 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, 2H), 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).

[0271] Examples 5 to 8: Synthesis of compounds 5 to 8

[0272] Compounds 5, 6, 7, and 8 of Examples 5, 6, 7, and 8, respectively, were prepared according to the same reactions as described in Steps 3 to 6 of Example 4, except that in Step 3 of Example 4, isocyanate 3,4-dichlorophenyl carbonate was replaced with reactant A of Table 2 below and in Step 4, 4-(bromomethyl)-3-fluorobenzoic acid methyl ester was replaced with reactant B of Table 2 below. The properties and yields of the products prepared in each of Steps 3 to 6 of Examples 5 to 8 are shown in Table 2, and the NMR data thereof are shown in Table 3. In Table 3 above, compounds 5, 6, 7, and 8 were prepared according to Examples 5, 6, 7, and 8, respectively.

[0273] [Table 2]

[0274]

[0275] Table 3

[0276]

[0277] Protocols for assaying and analyzing the activity of compounds of the invention

[0278] Experimental Example 1. Exploration of HDAC enzyme activity inhibition (in vitro)

[0279] Experiments were performed to identify the selectivity of the compound of the present application represented by Formula I for HDAC6 through experiments for HDAC1 and HDAC6 enzyme activity inhibition.

[0280] HDAC enzyme activity was measured using HDAC Fluorometric Assay Drug Discovery Kit (BML-AK511, 516) of Enzo Life Science, Inc. For the test for the HDAC1 enzyme activity, human recombinant HDAC1 (BML-SE456) was used as an enzyme source and Fluor de Lumi "SIRT1 (BNL-KI177)" as a substrate. Five-fold dilution of the compound was dispensed into a 96-well plate, and then 0.3 μg of the enzyme and 10 μM of the substrate were added to each well and subjected to reaction at 30°C for 60 minutes, after which Fluor de Lumi Developer II (BML-KI176) was added thereto and subjected to reaction for 30 minutes and ended. Thereafter, fluorescence value (Ex 360, Em 460) was measured using a multi-plate reader (Flexstation 3, Molecular Device). The experiment for HDAC6 enzyme was performed by using human recombinant HDAC6 (382180) of Calbiochem Inc. according to the same protocol as in the HDAC1 enzyme activity test method. For the final result value, IC 50 values were calculated using the GraphPad Prism 4.0 program.

[0281] Table 4

[0282]

[0283] As described in Table 4 above, it was confirmed from the results of testing the activity inhibition of HDAC1 and HDAC6 that the 1,3,4-oxadiazole oxetane derivative compound of the present application, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof showed excellent selective HDAC6 inhibitory activity of about 517-fold to about 1207-fold.

[0284] Experimental Example 2. Analysis of HDAC6 inhibitory activity and other HDAC isoform selectivity (in vitro)

[0285] The potency and selectivity of the HDAC6 inhibitory activity of the compound represented by Compound 5 (the compound of Example 5: Compound 5) and the compound represented by Compound 1 (the compound of Example 1: Compound 1) were confirmed at the enzyme level. This experiment was commissioned to Reaction biology Corp. (Malvern, PA, USA) and was performed according to an in-house established test method. Specifically, a serial dilution of the compound represented by Compound 5 or the compound represented by Compound 1 was dispensed into a plate, and then a substrate (i.e., RHK-K(Ac)-AMC) and an enzyme were added 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 a reaction. Thereafter, 50 mm 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 thereto and then subjected to a reaction. Then, a fluorescence signal was measured at Ex. 360 nm / Em. 460 nm to measure enzyme activity, and then the results thereof are shown in Table 5 below.

[0286] [Table 5]

[0287]

[0288] It was confirmed that the IC50 of HDAC6 of the compound represented by Compound 5 was 18.9 nM, while other HDAC isoforms were not inhibited at all. 50 It was confirmed that the IC50 of HDAC6 of the compound represented by Compound 1 was 60.0 nM, while other HDAC isoforms were not inhibited at all. 50 In other words, it was confirmed that the compound represented by 13608 and the compound represented by Compound 1 are compounds having excellent HDAC6 inhibitory activity and high selectivity for HDAC6 compared to other HDAC isoforms.

[0289] Experimental Example 3. Analysis of the effect on axonal migration of mitochondria (in vitro)

[0290] It has been reported that acetylation of tubulin, which is a component of microtubules, decreases in various degenerative brain diseases, and it is known that intracellular transport that occurs through microtubules is impaired due to various other intracellular dysfunctions.

[0291] HDAC6 is an enzyme that plays a role in removing acetylation of tubulin, and when the enzyme is inhibited, it is known that acetylation of tubulin increases to stabilize the microtubules and has a positive effect on intracellular transport and axonal transport.

[0292] Experiments were conducted for this example to confirm whether the compound represented by Compound 5 (the compound of Example 5: Compound 5) and the compound represented by Compound 1 (the compound of Example 1: Compound 1) according to the present application show an improving effect on the reduced mitochondrial relative velocity by treatment with a β-amyloid fragment (Aβ), which is a substance causing dementia in neurodegenerative brain diseases by selectively inhibiting HDAC6 activity to increase acetylation of tubulin, which is a major substrate of HDAC6, in the axon of neurons.

[0293] Specifically, the hippocampal tissue of mouse embryos obtained from ICR pregnant maternal mice was subjected to single cell suspension, and then the hippocampal neurons were cultured in a culture vessel coated with extracellular matrix for imaging for 7 days. After culturing for 7 days, the mouse hippocampal neurons were treated with Aβ at a concentration of 1 μM. After 24 hours, the resulting neurons were treated with the compound represented by Compound 5 and the compound represented by Compound 1 at a concentration of 0.3 μM for 3 hours, after which the migration of stained mitochondria was photographed at low speed by an automated cell photographing apparatus in order to measure the migration distance per unit time, thereby evaluating the degree of intracellular transport.

[0294] The images were photographed at 1 second intervals for 1 minute to measure the relative velocity of each mitochondrion per second. After setting the section in which the relative velocity of mitochondria was significantly reduced compared to the vehicle in the normal group in the β-amyloid treated group, the results were normalized to the vehicle group and are shown in Table 6 below.

[0295]

Table 6

[0296]

[0297] In Table 6 above, normal means a normal group treated only with a vehicle (0.5% DMSO) without treatment with a β-amyloid (Aβ), and β-amyloid vehicle means a group treated only with a β-amyloid and a vehicle (0.5% DMSO).

[0298] As shown in Table 6 above, it can be seen that the reduced relative velocity of mitochondria in the group of neurons treated with Aβ was increased by 3 hours of treatment with the compound represented by Compound 5 and the compound represented by Compound 1 compared to the normal group of neurons, and thus the reduced relative velocity of mitochondria in neurodegeneration including atrophy of the central nervous system, neurodegenerative diseases including neurodegenerative brain diseases, or hereditary neuropathy was improved by treatment with the compounds.

[0299] Accordingly, it has been confirmed that the compound of the present application exhibits an excellent effect of preventing and treating a nervous system atrophy including central nervous system atrophy, a neurodegenerative disease (including a neurodegenerative brain disease) or a hereditary neuropathy (including dementia and Alzheimer's disease).

[0300] Experimental Example 4. Analysis of the effect on the relative velocity of axons of mitochondria when treated at each concentration (in vitro)

[0301] The experiment for this example was performed to confirm whether the compound represented by Compound 5 according to the present application (the compound of Example 5: Compound 5) shows an improvement effect on the relative velocity of mitochondria decreased by treatment with a beta amyloid fragment (Aβ) which is a substance causing dementia in a neurodegenerative brain disease by selectively inhibiting HDAC6 activity to increase acetylation of tubulin which is a major substrate of HDAC6 in a neuron axon in a concentration-dependent manner.

[0302] Specifically, the hippocampal tissue of mouse embryos obtained from a pregnant maternal mouse was subjected to single cell suspension, and then the hippocampal neurons were cultured in a culture vessel coated with extracellular matrix for imaging for 7 days. After being cultured for 7 days, the mouse hippocampal neurons were treated with Aβ at a concentration of 1 μM. After 24 hours, the resulting neurons were treated with the compound represented by Compound 5 at concentrations of 0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30 μM for 3 hours, after which the migration of stained mitochondria was photographed at low speed by a confocal microscope in order to measure the migration distance per unit time, thereby evaluating the degree of intracellular transport.

[0303] The images were photographed at 1 second intervals for 1 minute to measure the relative velocity of each mitochondria per second. After setting a section in which the relative velocity of mitochondria was significantly decreased compared to the normal group in the beta amyloid treatment group, the normal group was normalized to 100% and the group treated only with beta amyloid was normalized to 0%, and the results are shown in Table 7 and Figure 1 below.

[0304]

Table 7

[0305]

[0306] In Table 7 above, normal means a normal group treated only with a solvent (0.5% DMSO) without treatment with a beta amyloid (Aβ), and solvent in beta amyloid means a group treated only with a beta amyloid and a solvent (0.5% DMSO).

[0307] As shown in Table 7 and Figure 1As shown in FIG. 5, it can be seen that the decreased mitochondrial relative velocity in the group of neurons treated with Aβ is significantly increased in a dose-dependent manner by 3-hour treatment with the compound represented by Compound 5, and thus the decreased mitochondrial relative velocity in neurodegeneration including central nervous system atrophy, neurodegenerative disease (including neurodegenerative brain disease) or hereditary neuropathy is improved by treatment with the compound.

[0308] Accordingly, it has been confirmed that the compounds of the present application exhibit excellent effects in preventing and treating neurodegeneration including central nervous system atrophy, neurodegenerative disease (including neurodegenerative brain disease) or hereditary neuropathy (including dementia and Alzheimer's disease).

[0309] Experimental Example 5. Analysis of duration of pharmacodynamic effect on mitochondrial axonal relative velocity (in vitro)

[0310] Experiments for this example were performed to confirm the duration of the improving effect on the decreased mitochondrial relative velocity by treatment with a β amyloid fragment (Aβ) by the compound represented by Compound 5 according to the present application (the compound of Example 5: Compound 5).

[0311] Specifically, the hippocampal tissue of mouse embryos obtained from pregnant maternal mice was subjected to single cell suspension, and then the hippocampal neurons were cultured in a culture vessel coated with extracellular matrix for imaging for 7 days. After 7 days of culture, the mouse hippocampal neurons were treated with Aβ at a concentration of 1 μM. After 24 hours, the resulting neurons were treated with the compound represented by Compound 5 at a concentration of 0.3 μM for 3 hours and replaced with fresh culture medium. Thereafter, in order to confirm the persistence of the pharmacodynamic effect for 2 hours to 24 hours when not treated with the compound, the migration of stained mitochondria was slowly photographed by confocal microscopy, and the migration distance per unit time was measured to evaluate the degree of intracellular transport.

[0312] The images were photographed for 1 minute at 1-second intervals to measure the relative velocity per second of each mitochondrion. After setting the section in which the mitochondrial relative velocity was significantly decreased compared to the vehicle of the normal group in the β amyloid-treated group, the results normalized to 100% of the vehicle and 0% of the group treated with β amyloid are shown in Table 8 and Figure 2 in FIG. 6.

[0313]

Table 8

[0314]

[0315] In Table 8 above, normal means a normal group treated with a solvent (0.5% DMSO) only and not treated with amyloid beta (Aβ), and amyloid beta solvent means a group treated with amyloid beta and a solvent (0.5% DMSO) only.

[0316] As shown in Table 8 and Figure 2 above, it has been confirmed that the decreased relative velocity of mitochondria in the group of neurons treated with Aβ compared to the normal group of neurons was significantly increased by 3-hour treatment with the compound represented by Compound 5, and the efficacy thereof was significantly sustained for up to 9 hours even after the compound was removed by replacing the culture medium with a new one.

[0317] Accordingly, it has been confirmed that the compound of the present application exhibits an excellent effect of preventing and treating neuroatrophy including central nervous system atrophy, neurodegenerative disease including neurodegenerative brain disease, or hereditary neuropathy including dementia and Alzheimer's disease.

[0318] Experimental Example 6. Analysis of the effect on the axonal relative velocity of mitochondria damaged in a tauopathy-like situation (in vitro)

[0319] In order to confirm the therapeutic effect of the compound represented by Compound 5 (the compound of Example 5: Compound 5) on degenerative brain disease, a cell model of tauopathy was prepared and treated with the compound of the present application, and then the experiment of this example was performed to confirm the duration of the improving effect on the decreased relative velocity of mitochondria.

[0320] It is known that tau binds to microtubules constituting nerve cells, thus helping to maintain the stability of the microtubules, and in pathological conditions such as tauopathy, tau is separated from the microtubules, and then the microtubules become unstable and adversely affect various cellular functions. Microtubules act as a pathway mediating the movement of various organelles, vesicles, and various substances required for cellular homeostasis in nerve cells, and under tauopathy conditions, such intracellular transport can not occur normally due to the instability of these microtubules. A test system was constructed using supercultured cells of mice having human tau with a P301L mutation that promotes aggregation and detachment from microtubules overexpressed, and in this test system, it was evaluated whether the compound represented by Compound 5 improves microtubule normalization and intracellular transport.

[0321] Specifically, the cerebral cortex tissue of mouse embryos obtained from pregnant mouse mothers was subjected to single cell suspension, and then transfected with a P301L mutant human tau protein expression vector using electroporation. After culturing in a cell incubator for 7 days, the resulting cells were treated with a compound represented by Compound 5 at concentrations of 0.01, 0.1, 0.3, 1, 3, 10 μM for 3 hours, after which the migration of stained mitochondria was photographed at low speed by confocal microscopy in order to measure the migration distance per unit time, thereby evaluating the degree of intracellular transport.

[0322] All results are expressed as mean ± standard error, and the effectiveness of the pharmacological effect was determined by statistical significance between the negative control group and each test substance group. Statistical analysis was performed using one-way ANOVA to confirm the homogeneity of dispersion, and when the p-value was less than 0.05 by Dunnett post test, it was determined to be statistically significant, and the test results are shown in Table 9 and Figure 3

[0323]

Table 9

[0324]

[0325] In Table 9 above, P301L_Tau(-) refers to a group treated only with a solvent (0.5% DMSO), which serves as a normal neuron group transfected with a control vector that does not cause tauopathy, and P301L_Tau(+) refers to a group treated only with a solvent (0.5% DMSO), which serves as a disease neuron group transfected with a P301L mutant human tau protein expression vector.

[0326] As shown in Table 9 and Figure 3 above, it can be seen that the reduced mitochondrial relative velocity in the P301L_Tau(+) group compared to the P301L_Tau(-) group was significantly increased in a dose-dependent manner by 3-hour treatment with a compound represented by Compound 5, and thus the reduced mitochondrial relative velocity in the tauopathy state was improved by treatment with the compound of the present application.

[0327] Accordingly, it has been confirmed that the compound of the present application exhibits an excellent effect of preventing and treating neurological atrophy including central nervous system atrophy, neurodegenerative diseases (including neurodegenerative brain diseases) or hereditary neuropathy (including tauopathy).

[0328] Experimental Example 7. Analysis of HDAC6 inhibitory activity and selectivity at the level of neurons (in vitro)

[0329] ​The HDAC6 inhibitory efficacy and selectivity of the compound represented by Compound 5 (the compound of Example 5: Compound 5) and the compound represented by Compound 1 (the compound of Example 1: Compound 1) were confirmed at the neuronal level.

[0330] The experiment of this example was performed to confirm whether the compound represented by Compound 5 and the compound represented by Compound 1 according to the present application selectively inhibit HDAC6 activity in neural cells to increase acetylation of tubulin, which is the main substrate of HDAC6.

[0331] Specifically, SH-SY5Y cells (human neuroblastoma) were attached to a culture dish and incubated in an incubator for 24 hours. After culturing for 1 day, the SH-SY5Y cells were treated with the compound represented by Compound 5 and the compound represented by Compound 1 at concentrations of 0.1, 0.3, and 1 μM for 3 hours. The cells were chemically lysed using RIPA buffer, and then the extracted proteins were arranged by each protein size on a polyacrylamide gel by SDS-PAGE technique. The proteins on the gel were transferred to a nitrocellulose membrane using electrophoresis, and the degree of expression was determined by allowing an antibody selectively bound to the proteins to be observed to react and visualized in the form of a band, and the results are shown in Figure 4 and Figure 5 .

[0332] In the above Figure 4 and Figure 5 , con refers to the results of cells not treated with the compound of the present application.

[0333] As can be confirmed from the above Figure 4 and Figure 5 , it was confirmed that the compound represented by Compound 5 and the compound represented by Compound 1 significantly increased acetylation of tubulin.

[0334] Therefore, it can be seen that the compound according to the present application exhibits an excellent effect of increasing acetylation of tubulin.

[0335] Experimental Example 8. Analysis of brain penetration of HDAC6-specific inhibitor (in vivo)

[0336] In order to observe the therapeutic effect of the compound on degenerative brain diseases, the compound exposed to blood needs to penetrate the blood-brain barrier (BBB) and reach neural cells in brain tissues.

[0337] The experiment for this example was performed to confirm whether the compound represented by Compound 5 (the compound of Example 5: Compound 5) and the compound represented by Compound 1 (the compound of Example 1: Compound 1) according to the present application are absorbed in the stomach, penetrate the blood-brain barrier (BBB), and reach neural cells in brain tissues when orally administered.

[0338] Specifically, ICR mice were orally administered once with the compound represented by Compound 5 at a concentration of 10 mg / kg, and blood and brain were collected after 0.5, 2, or 4 hours. Residual concentrations of the compound represented by Compound 5 were measured in the plasma of the harvested mouse blood and brain tissue by LC-MS / MS technique (see Rapid Commun. Mass Spectrom. 14, 1729-1735 (2000)). ICR mice were orally administered once with the compound represented by Compound 1 at a concentration of 50 mg / kg, and blood and brain were collected after 0.5, 2, or 4 hours. Residual concentrations of the compound represented by Compound 1 were measured in the plasma of the harvested mouse blood and brain tissue by LC-MS / MS technique and the results thereof are shown in Table 10.

[0339] All results are expressed as mean ± standard deviation of the mean, the unit of the compound concentration in the brain is ng / g, and the unit of the compound concentration in the plasma is ng / mL.

[0340]

Table 10

[0341]

[0342] As shown in Table 10 above, it has been confirmed that the compound represented by Compound 5 and the compound represented by Compound 1 have a B / P ratio of 0.3 or more, and thus have high brain penetration.

[0343] Accordingly, it can be seen that the compound according to the present application shows excellent brain penetration, and thus exhibits an excellent effect of preventing and treating neurological diseases (e.g., brain diseases, central nervous system atrophy, hereditary neurological diseases, etc.).

[0344] Experimental Example 9. Animal behavior evaluation (cognitive function evaluation)

[0345] The most prominent symptom of neurodegenerative diseases is cognitive dysfunction, and various assessment tools are used to evaluate the cognitive function of patients in actual clinical trials (MoCA, MMSE, CDR., ADAS-cog, etc.). In animal experiments, Y-maze, passive avoidance test, and water maze test are mainly used as cognitive / learning evaluation methods.

[0346] In order to confirm the therapeutic effect on cognitive dysfunction caused by neurodegeneration, the efficacy was confirmed in tauopathy mice (PS19) formed by transformation of tau, which is a causative protein of Alzheimer's disease and tauopathy.

[0347] Tauopathy is one of representative degenerative brain diseases, and tau (a pathogenic protein) is known to be associated with the formation of at least ten kinds of degenerative brain diseases. Tauopathy is a disease formed in a person having several mutations in MAPT (a pathogenic gene of tauopathy).

[0348] Tau is known to bind to microtubules constituting nerve cells, thus helping to maintain the stability of the microtubules, and under pathological conditions (e.g., tauopathy), tau is separated from the microtubules, and then the microtubules become unstable and adversely affect various cellular functions.

[0349] PS19 tauopathy mice are disease model animals showing clinical symptoms of patients suffering from tauopathy: for example, increase of tau and phosphorylated tau proteins in the brain, decrease of memory and cognitive functions, and decrease of motor functions, which are caused by overexpression of human P301S mutant tau protein that promotes tau aggregation and detachment from microtubules.

[0350] The compound of the present application was orally administered twice a day to PS19 tauopathy mice, and after 30 minutes from the administration of the test substance, the following behavioral tests were performed.

[0351] Y-maze test

[0352] The Y-maze test was performed to evaluate the ability to act in sequence in an experiment for determining short-term memory. The determination device consists of three parts each having a length of 42 cm, a width of 3 cm, a height of 12 cm, and a folding angle of 120°. The device consists of white polyethylene plastic, and the experiment was performed after setting three branches as A, B, and C, respectively. The experimental animals were placed, and the number of times each animal scored 1 point (actual alternation) by counting the times of complete entry into each branch even with its tail and sequential entry into each branch within 8 minutes. Alternation behavior was defined as entry into all three branches without repetition, and spontaneous alternation behavior was calculated by the following equation, and the result thereof is shown in Table 1. Figure 6 to Figure 8

[0353] In Table 1 above, Figure 6 to Figure 8 WT (littermate) is a mouse of a normal group orally administered with 0.5% methyl cellulose, PS19 (vehicle, littermate) is a PS19 mouse (control group) orally administered with 0.5% methyl cellulose, and PS19 + Compound 5 is a group of PS19 mice orally administered with a compound represented by Compound 5.

[0354] As Figure 6 ​As confirmed in Example 5, since compound 5 (compound of Example 5: Compound 5) was repeatedly administered (orally) to 12-week-old tauopathy (PS19) mice for 8 weeks, it was seen that the alternation behavior value was significantly increased compared to the control group (littermates administered with 0.5% methyl cellulose (vehicle, PS19)), thus showing that the decline in cognitive function caused by tauopathy was improved.

[0355] As Figure 7 As confirmed in Example 5, since compound 5 (compound of Example 5: Compound 5) was repeatedly administered (orally) to 12-week-old tauopathy (PS19) mice for 16 weeks, it was seen that the alternation behavior value was increased compared to the control group (littermates administered with 0.5% methyl cellulose (vehicle, PS19)), thus showing that the decline in cognitive function caused by tauopathy was improved.

[0356] As Figure 8 As confirmed in Example 5, since compound 5 (compound of Example 5: Compound 5) was repeatedly administered (orally) to 12-week-old tauopathy (PS19) mice for 16 weeks, it was seen that the alternation behavior value was increased compared to the control group (littermates administered with 0.5% methyl cellulose (vehicle, PS19)), thus showing that the decline in cognitive function caused by tauopathy was improved.

[0357] Thus, it was seen that the compounds of the present application exhibit excellent effects in preventing and treating nervous system atrophy including central nervous system atrophy, neurodegenerative diseases (including neurodegenerative brain diseases) or hereditary neuropathy (including tauopathy).

[0358] Experimental Example 10. Histopathological analysis

[0359] As in Experimental Example 9, after the test substance was administered to PS19 mice, histopathological analysis was performed on brain tissues after termination of administration of the substance. 0.5 hours after the final administration of Compound 5 (the compound of Example 5: Compound 5), the mice were anesthetized with isoflurane, and then the brain was extracted. For double immunofluorescence staining, brain tissue sections were washed twice with phosphate buffered saline (PBS) for 15 minutes, and reacted with a blocking solution in which bovine serum albumin (Sigma, USA) was mixed with PBS for 2 hours. After washing twice with PBS, the primary antibody was diluted with PBS containing Triton x-100 (0.3%) and normal donkey serum (5%), and reacted overnight at 4°C. Anti-phosphorylated-tau (Ser202, Thr205), monoclonal antibody (clone AT8, 1:300, Invitrogen, MN1020) and anti-NeuN polyclonal antibody (1:300, Invitrogen, PA5-78499) were used as the primary antibody. Thereafter, the tissues were washed with PBS, and reacted with secondary antibodies, Alexa488-labeled donkey anti-mouse IgG (1:500, Jackson Immunoresearch, 715-545-150, for phosphorylated-tau) and Cy3-labeled donkey anti-mouse IgG (1:500, Jackson Immunoresearch, 711-165-152, for NeuN) for 3 hours at room temperature. Then, the tissues were washed again with PBS, attached to a coated slide, dried well, and mounted with DAPI-containing mounting medium (Antifade Mounting Medium with DAPI, H-1200-10, Vector Laboratories) to prepare tissue samples.

[0360] The stained brain tissues were photographed using a confocal microscope, and the staining intensity of AT8 in each brain tissue region was measured using the image J program in the photographed area, and the results thereof are shown in Figure 9 to Figure 13

[0361] In the above Figure 9 to Figure 13 , WT (littermate) is a mouse of a normal group administered with 0.5% methyl cellulose, PS19 (vehicle, littermate) is a PS19 mouse (control group) administered with 0.5% methyl cellulose, and PS19+Compound 5 is a group of PS19 mice administered with a compound represented by Compound 5.

[0362] As shown in the above Figure 9 to Figure 13 ​​It was confirmed that, as a result of performing histopathological analysis on each brain region of the 12-week-old tauopathy (PS19) mice after repeated administration of the compound represented by Compound 5 for 8 weeks, the degree of hyperphosphorylation of the transformed tau (AT8, S202 / T205) was significantly higher in the control group mice (vehicle, littermates) than in the normal group mice (WT, littermates). On the contrary, it was confirmed that the animals administered with the compound represented by Compound 5 in the control group mice showed significantly improved hyperphosphorylation of tau.

[0363] Thus, it can be seen that the compound represented by Compound 5 improved the hyperphosphorylation of tau observed in the PS19 mice.

[0364] Thus, it can be seen that the compound of the present application exhibits an excellent effect of preventing and treating a nervous system atrophy including central nervous system atrophy, a neurodegenerative disease (including a neurodegenerative brain disease) or a hereditary neuropathy (including a tauopathy).

[0365] Experimental Example 11. Electrophysiological analysis

[0366] After administering the test substance to the PS19 mice as described in Experimental Example 9, electrophysiological analysis was performed on the brain tissue after terminating the administration of the substance. After 0.5 hours from the final administration of Compound 5 (the compound of Example 5: Compound 5), the mice were anesthetized, and then the brain was extracted. The extracted brain was sliced to a thickness of 300 μm to prepare a tissue slice containing the hippocampus, and was transferred to a recording chamber perfused with artificial cerebrospinal fluid (30 to 32°C) and continued.

[0367] Field excitatory postsynaptic potential (fEPSP) recordings were obtained using DAM80 amplifiers and WinLTP 2.10 software (University of Bristol) filtered at 1 kHz and sampled at 20 kHz. Recording pipettes with a resistance of 1-3 ΜΩ were filled with artificial cerebrospinal fluid. All fEPSPs were recorded on the Schaffer collateral pathway (CA3 to CA1 synapse) either by stimulation with two bipolar electrical stimulators (FHC) placed in the irradiated layer or by stimulation with 20 s inter-stimulus interval (ISI). Theta burst stimulation (TBS) was given as a stimulus train of 100 Hz (5 pulses, 20 times) at 5 Hz intervals. Confirmation of successful LTP induction was determined by statistical comparison of the mean fEPSP slope / amplitude 50-60 min after TBS to the mean fEPSP slope / amplitude 10 min before TBS. Analysis of data was performed using WinLTP 2.10 reanalysis software (University of Bristol). All data are expressed as mean ± standard error of the mean (SEM) and statistical analysis was performed using SPSS statistical version 21 (IBM). In comparing two groups, a Student t-test was used for significance testing, ANOVA for significance testing of group comparisons, with LSD post-hoc analysis, the results of which are shown in Figure 14 and Figure 15 .

[0368] Significance levels were set at p < 0.05. This means * or §, p < 0.01; ** or §§, p < 0.01; *** or §§§, p < 0.001.

[0369] In the above Figure 15 , 7 means the number of brain tissue sections and 2 means the number of mice that have undergone evaluation in terms of 7 / 2. In terms of 14 / 4, 14 means the number of brain tissue sections and 4 means the number of mice that have undergone evaluation.

[0370] In the above Figure 14 and Figure 15 , WT (littermate) is a mouse of a normal group orally administered with 0.5% methyl cellulose, PS19 (vehicle, littermate) is a PS19 mouse (control group) orally administered with 0.5% methyl cellulose, and PS19 + Compound 5 is a group of PS19 mice orally administered with a compound represented by Compound 5. As shown in the above Figure 14 and Figure 15It has been confirmed that, compared to the normal group, the magnitude of the long-term protein troponin (LTP) in the hippocampus of the control group was significantly reduced. This indicates the degeneration of neural pathways related to memory. Conversely, it has been confirmed that, through repeated (oral) administration of the compound represented by compound 5 to 12-week-old PS19 mice for 16 weeks, enhanced hippocampal long-term memory was significantly restored compared to the control group (livestock mice given 0.5% methylcellulose (solvent, PS19)).

[0371] Therefore, it can be seen that the compounds of the present invention exhibit excellent effects in preventing and treating nervous system atrophy, including central nervous system atrophy, neurodegenerative diseases (including neurodegenerative brain diseases), or hereditary neuropathies (including tau protein diseases).

[0372] Experimental Example 12. Animal Behavioral Assessment (Motor Function Assessment)

[0373] To confirm the therapeutic effects of the compound represented by 13524 (compound of Example 1: compound 1) and the compound represented by 5 (compound of Example 5: compound 5) on degenerative brain diseases, the compounds were administered to Yac128 mice, a Huntington's disease model mouse, and the improvement in motor function of the animals was subsequently evaluated.

[0374] Huntington's disease is a representative degenerative brain disease and a hereditary disorder that causes death from severe physical and mental impairment within 15 to 25 years after onset. Huntington's disease is an autosomal dominant disorder that develops in individuals with a mutation in which the CAG sequence is repeated 40 or more times in the HTT gene (responsible for Huntington's disease).

[0375] Yac128 is a mouse model of Huntington's disease, in which a human mutant HTT gene with 128 CAG repeat sequences is inserted. As a disease model animal, it exhibits clinical symptoms of Huntington's disease, such as the expression of human mutant HTT transcripts and proteins, striatum death, and deterioration of motor function (e.g., muscle strength and limb coordination).

[0376] Specifically, 6-month-old Yac128 mice were orally administered the compound represented by compound 1 at doses of 5 and 20 mg / kg twice daily for 8 weeks, and the compound represented by compound 5 was repeatedly orally administered at doses of 1, 3, 10, and 30 mg / kg twice daily for 12 weeks. Motor function was assessed at 4-week intervals, and the results are presented. Figure 16 to Figure 19 middle.

[0377] In the above Figure 16 to Figure 19In this study, all results are indicated as mean ± standard error, and efficacy was determined by statistical significance between the Yac128 mouse control group and each test substance group. For statistical analysis, homogeneity of dispersion was determined using ANOVA (one-way ANOVA for single assays and two-way ANOVA for repeated assays). Statistical significance was determined by Dunnett or Bonferroni post-hoc tests if the p-value was less than 0.05. Accelerated rotarod and grip strength tests were performed once daily for two days. Subsequently, based on the test results, mice were grouped using the Z-array method. Yac128 mice were divided into solvent-based administration groups and compound-based administration groups (compounds represented by compound 1: 5, 20 mg / kg; compounds represented by compound 5: 1, 3, 10, 30 mg / kg, administered orally), with 18 mice in each group. The test substances were administered orally twice daily for 8 and 12 weeks, and motor function was assessed twice daily at four-week intervals during the administration period.

[0378] Accelerated Rotor Test

[0379] The accelerated rotarod test (ROTA ROD, LE8205, Panlab) was conducted to assess motor coordination / motor function. Prior to this test, all test animals were placed on a bar accelerated from 4 rpm to 20 rpm three times a day for three days to train adaptation for approximately three weeks. Animals that took 180 seconds or more to fall from the bar were used for this test. In this experiment, the time it took for an animal to fall from the bar accelerated from 4 rpm to 40 rpm was measured within three minutes. The accelerated rotarod test was performed three times a day for two consecutive days, for a total of six times, and the maximum of the six measurements was used. The results are shown in... Figure 16 and Figure 17 middle.

[0380] In the above Figure 16 and Figure 17 In this context, WT (wild-type) refers to normal mice orally administered the solvent (0.5% methylcellulose), Yac128 refers to the control group of Huntington's disease model mice orally administered the solvent, Yac128+compound5 represents the group in which Yac128 mice were orally administered the compound represented by compound 5, and Yac128+compound1 represents the group in which Yac128 mice were orally administered the compound represented by compound 1.

[0381] As mentioned above Figure 16 As shown, the dropout latency (which was reduced compared to the normal mice in the Yac128 control group) was significantly increased by administering the compound represented by compound 1 at a dose of 20 mg / kg for 4 weeks.

[0382] In addition, such as Figure 17As shown in FIG. 2, it can be seen that the fall latency (which has been decreased in the Yac128 control group compared to the normal mice) is significantly increased at one week from the administration of the compound represented by Compound 5 at a dose of 3 mg / kg, and the fall latency is significantly increased in most cases during a longer administration period or at a higher concentration, and thus the loss of motor function, which is a symptom of Huntington's disease, is improved by the administration of the compound of the present application.

[0383] Therefore, it can be seen that the compound of the present application exhibits an excellent effect of preventing and treating a nervous system atrophy including central nervous system atrophy, a neurodegenerative disease (including a neurodegenerative brain disease) or a hereditary neuropathy (including Huntington's disease).

[0384] Grip strength test

[0385] A grip strength test (BIO-GS3, BIOSEB) was performed to evaluate muscle strength. The grip strength test evaluated the force of both forepaws of the mice using a bar. All experiments were performed by one person. When the mice gripped, the tail was gently pulled to have grip strength, and then pulled at an inclination of 15° to measure the maximum tension. The grip strength test was performed a total of ten times for two consecutive days, five times a day, and the average of these values was used, and the results thereof are shown in FIGS. 1, 3, 5, 7, 9, 11, 13, 15, 17 and 19. Figure 18 and Figure 19

[0386] In the above Figure 18 and Figure 19 , WT (wild type) means normal mice to which a vehicle was administered, Yac128 means a control group of Huntington's disease model mice to which a vehicle was administered, Yac128 + Compound 5 indicates a group in which a compound represented by Compound 5 was administered to Yac128 mice, and Yac128 + Compound 1 represents a group in which a compound represented by Compound 1 was administered to Yac128 mice.

[0387] As shown in the above Figure 18 , by administering a compound represented by Compound 1 for 8 weeks, the grip strength (which was decreased in the Yac128 control group compared to normal mice) was significantly increased in a dose-dependent manner.

[0388] Further, as shown in the above Figure 19 , it can be seen that, by administering a compound represented by Compound 5 for 12 weeks, the grip strength (which was decreased in the Yac128 control group compared to normal mice) was significantly increased, and was significantly increased in all administration groups at all concentrations, and thus the decrease in muscle strength, which is a symptom of Huntington's disease, was improved by the administration of the compound.

[0389] ​Thus, it can be seen that the compounds of the present application exhibit excellent effects in preventing and treating nervous system atrophy including central nervous system atrophy, neurodegenerative diseases including neurodegenerative brain diseases, or hereditary neuropathy including Huntington's disease.

[0390] Experimental Example 13. Analysis of the effect on the increased axonal relative velocity of damaged mitochondria in Charcot-Marie-Tooth disease (CMT, HMSN, hereditary motor and sensory neuropathy) (in vitro)

[0391] To confirm the therapeutic effect of the compound represented by Compound 5 (the compound of Example 5: Compound 5) and the compound represented by Compound 1 (the compound of Example 1: Compound 1) on CMT disease, experiments for this example were performed to confirm the ameliorating effect on the reduced mitochondrial movement velocity in neuronal axons isolated from the dorsal root ganglion of a 9-month-old MFN2 mutant mouse having CMT type 2A Charcot-Marie-Tooth disease induced according to treatment with the compound of the present application.

[0392] After culturing for 3 days in a cell incubator, mouse neuronal cells obtained by isolating dorsal root ganglion (DRG) from a 9-month-old MFN2 mutant mouse were treated with Compound 5 and Compound 1 at concentrations of 100 nM and 300 nM for 3 hours, and then the migration of stained mitochondria was slowly photographed by confocal microscopy to measure the migration distance per unit time, thereby evaluating the degree of intracellular transport.

[0393] The results are expressed as the mean ± standard error, and the effectiveness of the pharmacological effect was determined by statistical significance between the negative control group and each test substance group. Statistical analysis was performed using one-way ANOVA to confirm the homogeneity of dispersion, and it was determined to have statistical significance when the p-value by Dunnett's post-hoc test was less than 0.05.

[0394] In addition, in the MFN R94Q After setting a section in which the relative velocity of mitochondria was significantly reduced compared to the normal group (WT DRG) in mouse DRG, normalization to 100% of the normal group and MFN R94Q Mouse DRG 0%, and the test results are shown in Table 11 and Figure 20 below.

[0395] In Table 11 above, WT means a group in which a vehicle (0.5% methylcellulose) was administered to a normal mouse, and MFN R94Q Mouse DRG means a group in which only a vehicle (0.5% methylcellulose) was administered to a MFN2 mutant mouse (a group having induced CMT type 2A Charcot-Marie-Tooth disease).

[0396]

Table 11

[0397]

[0398] As shown in Table 11 and Figure 20 As shown in Table 11 and

[0399] Therefore, it has been confirmed that the compounds of the present application exhibit excellent effects in preventing and treating a nervous system atrophy including central nervous system atrophy, a neurodegenerative disease including neurodegenerative brain disease, or a hereditary neuropathy including Charcot-Marie-Tooth disease.

[0400] Experimental Example 14. Confirmation of therapeutic effect on CMT

[0401] CMT is the most common type of hereditary peripheral neuropathy, which is caused by mutations in proteins that make up nerves. To date, over 1000 mutations have been identified from about 90 genes (Timmerman et al., (2014) Genes 5:13-32). When Charcot-Marie-Tooth (CMT) disease occurs, progressive degeneration of peripheral nerves leads to atrophy of muscles affected by nerve distribution, and thus patients exhibit symptoms of gradual atrophy of their hand and foot muscles and deformed hands and feet. CMT is extremely diverse and complex in genetics and clinics, and it is known that its symptoms vary from near normal state to wheelchair-dependent state depending on the type of mutation. CMT mainly occurs in adolescents and occurs in one in every 2500 people (Krajewski et al., (2000) Brain 123:1516).

[0402] CMT belongs to an orphan disease as a hereditary peripheral neuropathy. However, its prevalence is equivalent to one in every 2500 people. There are about 20,000 patients in Korea and 2,800,000 patients worldwide. So far, therapies for CMT have been limited to rehabilitation, aids, pain control, surgical therapy, etc., but a successful therapeutic agent has not yet been developed. Therefore, there is a great need for the development of a therapeutic agent for CMT.

[0403] For example, with respect to CMT, which is the most common type of inherited motor and sensory neuropathy, large-scale clinical trials based on ascorbic acid, it has been proven through experiments based on culturing lemmocytes and dorsal root ganglion cells together that ascorbic acid is an essential substance for myelination in the peripheral nervous system, but this experiment could not prove effectiveness (Pareyson et al., (2011) 10(4):3205). In particular, with respect to certain types of diseases such as CMT1X, CMT2A, etc., the genes causing the diseases are highly expressed in the central nervous system. In fact, it has been confirmed that about 10% of patients have symptoms such as atrophy of certain brain tissues including the optic nerve, and thus the importance of the drug acting on the central nervous system is also mentioned.

[0404] As described above, the compound of the present application can pass through the blood-brain barrier, and thus can exhibit a therapeutic effect on Charcot-Marie-Tooth disease (CMT) related to the peripheral nervous system (PNS) as well as a therapeutic effect on Charcot-Marie-Tooth disease (CMT) related to the central nervous system (CNS).

[0405] Therefore, in the following studies for two types of CMT disease models, it has been confirmed that the compound of the present application can be advantageously used for the prevention and treatment of CMT by significantly improving the motor and sensory functions of CMT mice (rotarod and balance beam tests).

[0406] 14.1 Evaluation of motor and sensory functions

[0407] In inherited neurological diseases such as Charcot-Marie-Tooth disease (CMT), damage to the nervous system causes gait disturbance, sensory loss, decreased limb coordination, etc. In clinical studies of patients, various assessment tools such as walking 6 meters, evaluating sensory function according to the position of the arms and legs, tying a button, etc. are used to assess such functional defects. In addition, in animal experiments, methods such as rotarod test, balance beam test, grip strength test, etc. are mainly used to evaluate symptoms caused by such damage to the nervous system.

[0408] In the corresponding studies, in order to confirm the therapeutic effect of the compound represented by Compound 5 (the compound of Example 5: Compound 5) on the motor and sensory functions of CMT disease, the compound was administered to two CMT disease model mice (CX32 deletion mice, MFN2 mutant mice), and then the improvement effect on the corresponding functions of the animals was evaluated.

[0409] 14.1.1 CX32 Deletion Mouse Study Results

[0410] First, 5-month-old CX32 knockout mice were orally administered Compound 5 represented by Compound 5 at 10 mg / kg twice a day for 5 months, and motor and sensory functions were evaluated during the administration period. The results thereof are shown in Table 1. Figure 21 to Figure 22

[0411] In the following Figure 21 to Figure 22

[0412] For the grouping of the respective studies, the animals were divided into groups based on the body weight of the animals before administration, values derived from the constant rotarod and balance beam test results according to the Z-array method, as shown in Table 12 below. The mice were orally administered with a vehicle (0.5% methylcellulose) or Compound 5 represented by Compound 5 at 10 mg / kg according to the defined groups.

[0413] Male CX32 knockout mice used for the respective studies were provided with a standard diet (Central Lab Animal, Inc.) and water ad libitum and were housed in a controlled environment with temperature (22 ± 2℃), humidity (44% to 56%), and a 12-hour light-dark cycle. All experimental procedures were approved and conducted in accordance with the Institutional Animal Care and Use Committee (IACUC) of the Korea Center for Disease Control (CKD) (with approval number: S-22_016).

[0414]

Table 12

[0415]

[0416] Constant Rotarod Test

[0417] The rotarod test (LE 8205, Panlab) was performed to evaluate forced motor activity and coordination function. For acclimation, all test animals were acclimated to the test apparatus for a three-day session at 8 rpm five times a day, and for a two-day session at 12 rpm five times a day. Animals that met the category of fall latency of 100 seconds to 180 seconds were used for further experiments (about 80% of animals met the category). The fall latency was measured three times at a fixed speed of 12 rpm for three minutes each time. The rotarod test was repeated three times for each experiment and the maximum value of the three measurement values was used as the test result (fall latency). ​​

[0418] In the above Figure 21 , the normal group (wild type, WT) means normal mice to which a vehicle was orally administered, the CX32 knockout mice means a control group of CMT disease model mice to which a vehicle was orally administered, and the CX32 knockout mice + Compound 5 represents a group in which a compound represented by Compound 5 was orally administered to the CMT disease mice in which CX32 was knocked out (CX32 knockout mice).

[0419] As a result, as shown in Figure 21 , it has been confirmed that the compound of the present application exhibits an effect of significantly increasing the fall latency of the CX32 knockout mice.

[0420] Accordingly, it has been confirmed that the compound of the present application exhibits an excellent effect of preventing and treating atrophy of the nervous system, neurodegenerative diseases including neurodegenerative brain diseases, or hereditary neuropathies including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS).

[0421] Balance beam test

[0422] The balance beam test was performed to determine motor coordination function and limb-specific sensation. A beam (1.2 cm wide, 0.6 cm high, and 1.0 m long) was fixed with a slope of 9° (45 cm high from the starting point and 60 cm high from the end point). At the starting point, the mice were stimulated with a light of 60 W and the end point was equipped with a light-free dark box so that the mice could feel relaxed. All experimental animals were acclimated to the environment for 30 minutes before evaluation under the same conditions as the experimental conditions. The mice were placed at the starting point to walk toward the end point, and the number of slips after starting was determined. Before the test, the mice were trained three times a day for two days, and the results on the 3rd day were used for grouping. Each experiment was independently evaluated by two individuals, and the results thereof are shown in Figure 22 .

[0423] In the above Figure 22 , the normal group (wild type, WT) means normal mice to which a vehicle was orally administered, Veh means a control group of CMT disease model mice to which a vehicle was orally administered, and Compound 5 represents a group in which a compound represented by Compound 5 was orally administered to the CMT disease mice in which CX32 was knocked out (CX32 knockout mice).

[0424] As shown in Figure 22 , it has been confirmed that the compound of the present application exhibits an effect of significantly reducing the slip count and the beam crossing time.

[0425] Accordingly, it has been confirmed that the compounds of the present application exhibit excellent effects in preventing and treating neurodegenerative diseases, including neurodegenerative brain diseases, or inherited neuropathies, including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS).

[0426] 14.1.2 MFN2 mutant mouse research results

[0427] Six-month-old MFN2 mutant mice were orally administered with Compound 3 represented by Compound 5 at 10 mg / kg twice a day for 3 months, and motor and sensory functions were evaluated during the administration period, and the results thereof are shown in Figure 23 to Figure 24 The representation and statistical processing method for the corresponding results were performed in the same manner as the research of the CX32 deletion mice of 14.1.1 above.

[0428] For grouping for the corresponding research, the animals were grouped into each group based on the body weight of the animals before administration, values derived from the results of the accelerating rotarod and balance beam tests according to the Z-array method, as shown in Table 13 below. The mice were orally administered with a vehicle (0.5% methylcellulose) or 10 mg / kg of Compound 5 according to the defined groups.

[0429] Male MFN2 mutant mice for the corresponding research were provided with a standard diet (Central Lab Animal, Inc.) and water without restriction and were raised in a controlled environment with temperature (22 ± 2℃), humidity (44% to 56%), and a 12-hour light-dark cycle. All experimental procedures were approved and conducted in accordance with the Institutional Animal Care and Use Committee (IACUC) of the Korea CKD Experimental Animal Center (with approval number: S-22_012).

[0430]

Table 13

[0431]

[0432] Accelerating Rotarod Test

[0433] The accelerating rotarod test (LE8205, Panlab) was performed to evaluate motor coordination function / motor function. Prior to the test, all test animals were placed on a rod accelerating from 4 rpm to 20 rpm three times a day for three days to train for adaptation for about three weeks, and animals that took 180 seconds or more to fall off the rod were used for the test. In this experiment, the time for the animals to fall off the rod accelerating from 4 rpm to 40 rpm in three minutes was measured. The accelerating rotarod test was performed a total of three times a day, and the maximum value among three measured values was used. The results thereof are shown in Figure 23

[0434] ​In the above Figure 23 , the normal group (wild type, WT) means normal mice to which a vehicle was orally administered, Veh means a control group of CMT disease model mice in which a vehicle (0.5% methyl cellulose) was orally administered to MFN2 mutant mice, and Compound 5 represents a group in which a compound represented by Compound 5 was orally administered to MFN2 mutant mice.

[0435] As a result, as shown in Figure 23 , it has been confirmed that the compound of the present application exhibits an effect of significantly increasing the fall latency.

[0436] Accordingly, it has been confirmed that the compound of the present application exhibits an excellent effect of preventing and treating atrophy of the nervous system, a neurodegenerative disease (including a neurodegenerative brain disease) or a hereditary neuropathy (including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS), for example).

[0437] Balance beam test

[0438] The balance beam test was performed to determine motor coordination function and limb-specific sensation, and the research method was carried out in the same manner as the research of the CX32 deletion mouse of 14.1.1 above, and the results thereof are shown in Figure 24 .

[0439] In the above Figure 24 , the normal group (wild type, WT) means normal mice to which a vehicle was orally administered, Veh means a control group of CMT disease model mice in which a vehicle (0.5% methyl cellulose) was orally administered to MFN2 mutant mice, and Compound 5 represents a group in which a compound represented by Compound 5 was orally administered to MFN2 mutant mice.

[0440] As shown in the above Figure 24 , it has been confirmed that the compound of the present application exhibits an effect of significantly reducing the slip count.

[0441] Accordingly, it has been confirmed that the compound of the present application exhibits an excellent effect of preventing and treating atrophy of the nervous system, a neurodegenerative disease (including a neurodegenerative brain disease) or a hereditary neuropathy (including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS), for example).

[0442] 14.1.3 CMT2A mouse research results

[0443] CMT2A mice (mutant Mfn2 R94QCompound 3 represented by Compound 5 of Example 5 was administered orally at 10 mg / kg twice a day for 3 months, and motor and sensory functions were evaluated during the administration period, and the results thereof are shown in Table 14. Figure 25 to Figure 26 The representation and statistical processing method for the corresponding results were performed in the same manner as in the study of the CX32 deletion mouse of 14.1.1 above.

[0444] For the grouping of the corresponding study, the animals were grouped based on the body weight of the animals before administration, values derived from the results of the accelerating rotarod and balance beam tests according to the Z-array method, as shown in Table 14 below. The mice were orally administered with a vehicle (0.5% methyl cellulose) or 10 mg / kg of Compound 5 according to the defined groups.

[0445] Male CMT2A mice for the corresponding study were provided with a standard diet (Central Lab Animal, Inc.) and water ad libitum and were housed in a controlled environment with temperature (22 ± 2℃), humidity (44% to 56%), and a 12-hour light-dark cycle. All experimental procedures were approved and conducted in accordance with the Institutional Animal Care and Use Committee (IACUC) of the Korea Center for Disease Experimental Animals (with approval number: S-22_012).

[0446] [Table 14]

[0447]

[0448] Accelerating Rotarod Test

[0449] The accelerating rotarod test (ROTAROD, LE8205, Panlab) was performed to evaluate motor coordination function / motor function. Prior to the test, all test animals were placed on a rod accelerating from 4 rpm to 20 rpm three times a day for three days to train for about three weeks, and animals that took 180 seconds or more to fall off the rod were used for the test. In this experiment, the time for the animals to fall off the rod accelerating from 4 rpm to 40 rpm in three minutes was measured. The accelerating rotarod test was performed a total of three times a day, and the maximum value between three measurement values was used. The accelerating rotarod test was performed for 3 months, and the AUC data (period x measurement value in the rotarod test) obtained using the prism file are shown in Table 14. Figure 25

[0450] In Table 14 above, Figure 25 The results, as shown in Table 14, indicate that the motor coordination function / motor function of the CMT2A mice was significantly improved in the group in which Compound 5 represented by Compound 5 of Example 5 was administered orally at 10 mg / kg twice a day for 3 months, compared to the control group in which a vehicle (0.5% methyl cellulose) was administered orally to the CMT2A mice.

[0451] The results, as shown in Table 14, indicate that the motor coordination function / motor function of the CMT2A mice was significantly improved in the group in which Compound 5 represented by Compound 5 of Example 5 was administered orally at 10 mg / kg twice a day for 3 months, compared to the control group in which a vehicle (0.5% methyl cellulose) was administered orally to the CMT2A mice.​Figure 25 As shown in Table 2, it was confirmed that the compound of the present application exhibited an effect of significantly increasing the fall latency.

[0452] Accordingly, it was confirmed that the compound of the present application exhibited an excellent effect of preventing and treating neurodegenerative diseases including neurodegenerative brain diseases or genetic neuropathies including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS).

[0453] Balance beam test

[0454] The balance beam test was performed to determine motor coordination function and limb-specific sensation, and the research method was performed in the same manner as the research of CX32 deletion mice of 14.1.1 above, and the results thereof are shown in Table 2. Figure 26

[0455] In Table 2 above, Figure 26 the normal group (wild type, WT) means normal mice to which a vehicle was orally administered, Veh (TG) means a CMT disease model mouse control group in which a vehicle (0.5% methyl cellulose) was orally administered to CMT2A mice, and Compound 5 (TG) represents a group in which a compound represented by Compound 5 (Example 5) will be orally administered to CMT2A mice.

[0456] As shown in Table 2 above, Figure 26 it was confirmed that the compound of the present application (e.g., Compound 5 of Example 5) exhibited an effect of significantly reducing the slip count.

[0457] Accordingly, it was confirmed that the compound of the present application exhibited an excellent effect of preventing and treating neurodegenerative diseases including neurodegenerative brain diseases or genetic neuropathies including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS).

[0458] 14.1.4 CMT1X mouse research results

[0459] 5-month-old CMT1X mice (Gjb1 KO) were orally administered Compound 6 represented by Compound 5 at 10 mg / kg twice a day for 6 months, and motor and sensory functions were evaluated during the administration period, and the results thereof are shown in Table 3. Figure 27 to Figure 28 The representation and statistical processing method for the corresponding results were performed in the same manner as the research of CX32 deletion mice of 14.1.1 above.

[0460] ​For the grouping of the respective study, the animals were divided into groups based on the body weight of the animals before administration, values derived from the results of the accelerating rotarod and balance beam tests according to the Z-array method, as shown in Table 15 below. The mice were orally administered with vehicle (0.5% methylcellulose) or 10 mg / kg of Compound 5 of Example 5 according to the defined groups.

[0461] The male CMT1X mice used for the respective study were provided with a standard diet (Central Lab Animal, Inc.) and water ad libitum and housed in a controlled environment with temperature (22 ± 2℃), humidity (44% to 56%), and a 12-hour light-dark cycle. All experimental procedures were approved and conducted in accordance with the Institutional Animal Care and Use Committee (IACUC) of the CKD Experimental Animal Center of Korea (with approval number: S-22_012).

[0462] [Table 15]

[0463]

[0464] Constant Rotarod Test

[0465] A constant rotarod test (ROTAROD, LE8205, Panlab) was performed to evaluate motor coordination function / motor function. Prior to the test, all test animals were placed on a rod rotating at 10 rpm three times a day for three days to train for adaptation for about three weeks, and animals that took 180 seconds or more to fall off the rod were used for the test. In this experiment, the time for the animals to fall off the rod within three minutes was measured. The constant rotarod test was performed a total of three times a day, and the maximum value among three measurement values was used. The results thereof are shown in Figure 27 .

[0466] In the above Figure 27 , the normal group (wild type, WT) refers to normal mice to which vehicle was orally administered, Veh (TG) means a CMT disease model mouse control group in which vehicle (0.5% methylcellulose) was orally administered to CMT1X mice, and Compound 5 (TG) represents a group in which a compound represented by Compound 5 (Example 5) was orally administered to CMT1X mice.

[0467] As a result, as shown in Figure 27 , it has been confirmed that the compound of the present application exhibits an effect of significantly increasing the fall latency.

[0468] Accordingly, it has been confirmed that the compounds of the present application exhibit excellent effects in preventing and treating atrophy of the nervous system, neurodegenerative diseases including neurodegenerative brain diseases, or hereditary neuropathy including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS).

[0469] Balance beam test

[0470] The balance beam test was performed to determine motor coordination function and limb-specific sensation, and the research method was performed in the same manner as the research of the CX32 deletion mouse of 14.1.1 above, and the results thereof are shown in Figure 28 .

[0471] In the above Figure 28 , the normal group (wild type, WT) means a normal mouse to which a vehicle was orally administered, Veh (TG) means a CMT disease model mouse control group in which a vehicle (0.5% methyl cellulose) was orally administered to a CMT1X mouse, and Compound 5 (TG) represents a group in which a compound represented by Compound 5 (Example 5) was orally administered to a CMT1X mouse.

[0472] As shown in the above Figure 28 , it has been confirmed that the compounds of the present application (e.g., Compound 5 of Example 5) exhibit an effect of significantly reducing the slip count.

[0473] Accordingly, it has been confirmed that the compounds of the present application exhibit excellent effects in preventing and treating atrophy of the nervous system, neurodegenerative diseases including neurodegenerative brain diseases, or hereditary neuropathy including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS).

[0474] 14.1.5 CMT1A mouse research results

[0475] CMT1A mice (hPMP22 3-4 copies) of 6 weeks and a half were orally administered with Compound 6 represented by Compound 5 at 10 mg / kg twice a day for 6 weeks, and motor and sensory functions were evaluated during the administration period, and the results thereof are shown in Figure 29 to Figure 30 . The representation and statistical processing method for the corresponding results were performed in the same manner as the research of the CX32 deletion mouse of 14.1.1 above.

[0476] For the grouping of the respective study, the animals were divided into groups based on the body weight of the animals before administration, values derived from the results of the accelerating rotarod and balance beam tests according to the Z-array method, as shown in Table 16 below. The mice were orally administered with vehicle (0.5% methylcellulose) or 10 mg / kg of Compound 5 of Example 5 according to the defined groups.

[0477] The male CMT1A mice used for the respective study were provided with standard diet (Central Lab Animal, Inc.) and water ad libitum and housed in a controlled environment with temperature (22 ± 2℃), humidity (44% to 56%), and 12-hour light-dark cycle. All experimental procedures were approved and conducted in accordance with the Institutional Animal Care and Use Committee (IACUC) of the CKD Experimental Animal Center in Korea (with approval number: S-22_012).

[0478] [Table 16]

[0479]

[0480] Constant Rotarod Test

[0481] A constant rotarod test (ROTAROD, LE8205, Panlab) was performed to evaluate motor coordination function / motor function. Prior to the test, all test animals were placed on a rod rotating at 10 rpm three times a day for three days to train for about three weeks, and animals that took 180 seconds or more to fall off the rod were used for the test. In this experiment, the time for the animals to fall off the rod within three minutes was measured. The constant rotarod test was performed a total of three times a day, and the maximum value among three measurement values was used. The results thereof are shown in Table 17. Figure 29

[0482] In the above Figure 29 , the normal group (wild type, WT) refers to normal mice to which vehicle was orally administered, Veh (TG) means a CMT disease model mouse control group in which vehicle (0.5% methylcellulose) was orally administered to CMT1A mice, and Compound 5 (TG) represents a group in which a compound represented by Compound 5 (Example 5) was orally administered to CMT1A mice.

[0483] As a result, as shown in Table 17, Figure 29 , it has been confirmed that the compound of the present application (for example, Compound 5 of Example 5) exhibits an effect of significantly increasing the fall latency.

[0484] ​Accordingly, it has been confirmed that the compounds of the present application exhibit excellent effects in preventing and treating atrophy of the nervous system, neurodegenerative diseases including neurodegenerative brain diseases, or hereditary neuropathy including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS).

[0485] Balance beam test

[0486] The balance beam test was performed to determine motor coordination function and limb-specific sensation, and the research method was performed in the same manner as the research of the CX32 deletion mouse of 14.1.1 above, and the results thereof are shown in Figure 30 .

[0487] In the above Figure 30 , the normal group (wild type, WT) means normal mice to which a vehicle was orally administered, Veh (TG) means a CMT disease model mouse control group in which a vehicle (0.5% methyl cellulose) was orally administered to CMT1A mice, and Compound 5 (TG) represents a group in which a compound represented by Compound 5 (Example 5) was orally administered to CMT1A mice.

[0488] As shown in the above Figure 30 , it has been confirmed that the compounds of the present application (e.g., Compound 5 of Example 5) exhibit effects of significantly reducing the slip count.

[0489] Accordingly, it has been confirmed that the compounds of the present application exhibit excellent effects in preventing and treating atrophy of the nervous system, neurodegenerative diseases including neurodegenerative brain diseases, or hereditary neuropathy including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS).

[0490] 14.2 Nerve conduction study (NCS)

[0491] Through this experiment, the efficacy of the compounds of the present application was evaluated by confirming the effects of the compounds of the present application on the nerve conduction velocity of animals.

[0492] The animals used in the nerve conduction study were the animals studied in Section 14.1.2, and the nerve conduction study was performed after the administration of the compounds or a vehicle to male 6-month-old MFN2 mutant mice for 3 months.

[0493] Animals were anesthetized with isoflurane (USP Terrel, Piramal Critical Care, Inc., NDC 66794-017-25) in 30% oxygen (Daehan gas) and 70% nitrogen (Daehan gas). In this state, electrophysiological recordings were evaluated by electrical conduction in the tail nerve associated with sensory nerve conduction in the peripheral nervous system (PNS). Nicolet Viking Quest was used for the corresponding nerve conduction study (NCS). Sensory neuron action potential (SNAP) amplitude and sensory neuron conduction velocity (SNCV) were measured.

[0494] Data are expressed as mean ± SEM, and statistical significance between the group treated with the compound of the present application and the vehicle group was analyzed for comparison of three or more groups using one-way ANOVA (post-hoc analysis using Dunnett's test). All statistical analyses were performed using GraphPad Prism (version 9.0) and the results thereof are shown in Figure 31

[0495] In the above Figure 31 , the normal group (wild type, WT) refers to normal mice to which a vehicle was orally administered, Veh means a control group of CMT disease model mice in which a vehicle (0.5% methyl cellulose) was orally administered to MFN2 mutant mice, and Compound 5 represents a group in which a compound represented by Compound 5 was orally administered to MFN2 mutant mice.

[0496] As Figure 31 shown in the above, it has been confirmed that the compound of the present application exhibits an effect of significantly improving SNAP and SNCV, and thus is advantageously used for the prevention and treatment of CMT by improving nerve conduction velocity.

[0497] Accordingly, it has been confirmed that the compound of the present application exhibits an excellent effect of preventing and treating atrophy of the nervous system, neurodegenerative diseases (including neurodegenerative brain diseases) or hereditary neuropathies (including Charcot-Marie-Tooth disease such as Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) (CMT) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS) (CMT)).

[0498] 14.3. Histopathological analysis

[0499] Through this experiment, an attempt was made to confirm the effect of the compound of the present application on the axon size of sciatic nerve fibers.

[0500] 14.3.1 Results of CX32 deletion mouse study

[0501] ​Animals used in histopathological analysis were animals studied in Section 14.1.1, and the histopathological analysis was performed after administration of the compound or vehicle to male 5-month-old CX32 knockout mice for 5 months.

[0502] After the final treatment with the compound of the present application 5 (10 mg / kg), the sciatic nerve was collected at 0.5 hour and fixed in a 2.5% glutaraldehyde solution (340855, Sigma) overnight. The fixed sample was transferred to the Department of Pathology in University of Ulsan for semi-thin sectioning and toluidine blue (T3260, Sigma) staining.

[0503] The fixed sample was processed for image analysis (see Acta Neuropathologica Communications Volume 7, Article Number: 144 (2019), Sele et al.) and then 0.5 pm sections were prepared and stained with toluidine blue.

[0504] Histological evaluation was performed under an optical microscope. Pathological changes, including demyelination, remyelination, abnormally thin myelin sheath, and axonal morphological changes, were studied from the sections. Finally, the diameter of the axon was analyzed using image J software, and the results thereof are shown in Figure 32 .

[0505] In the above Figure 32 , the normal group (wild type, WT) refers to a normal mouse to which a vehicle was orally administered, Veh means a CMT disease model mouse control group in which a vehicle was orally administered to a CX32 knockout mouse, and Compound 5 represents a group in which a compound represented by Compound 5 was orally administered to a CMT disease mouse (CX32 knockout mouse) in which CX32 was knocked out.

[0506] Data are expressed as mean ± SEM. For comparison of three or more groups, statistical significance between the group treated with the compound of the present application and the vehicle group was analyzed using one-way ANOVA (post-hoc analysis using Dunnett's test). Statistical analysis was performed using GraphPad Prism (version 9.0).

[0507] As shown in Figure 32 , it has been confirmed that the compound of the present application significantly improves the increase in the axon size of the sciatic nerve fiber.

[0508] Thus, it has been confirmed that the compounds of the present application exhibit excellent effects in preventing and treating neurodegenerative diseases, including neurodegenerative brain diseases, or hereditary neuropathies, including Charcot-Marie-Tooth disease associated with the peripheral nervous system (PNS) (CMT) and Charcot-Marie-Tooth disease associated with the central nervous system (CNS) (CMT).

Claims

1. A compound represented by the following Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: [Formula I] wherein in the above Formula I, R1is H, F, Cl, Br, or I; R2and R3are each independently F, Cl, Br, or I.

2. The compound represented by Formula I according to claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R2and R3are each independently F or Cl.

3. The compound represented by Formula I according to claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound represented by the above Formula I is a compound represented by the following Formula II: [Formula II] wherein in the above Formula II, R1is H, F, Cl, Br, or I; R2and R3are each independently F, Cl, Br, or I. X1to X4are each independently N or CR x wherein three or more of X1to X4cannot be N at the same time, and R x is -H, F, Cl, Br or I; R1is -CX a H2, -C(X a )2H or -C(X a )3, wherein Xa is F, Cl, Br or I; and 4. The compound represented by Formula I according to claim 1 or 3, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound represented by the above Formula I is a compound represented by the following Formula II-1, II-2, II-3, or II-4: [Formula II-1] [Formula II-2] [Formula II-3] [Formula II-4] wherein in the above Formula II-1, II-2, II-3, or II-4, R1is H, F, Cl, Br, or I; R2and R3are each independently F, Cl, Br, or I in each formula.

5. The compound represented by Formula I according to claim 4, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: in the above Formula II-1, II-2, II-3, or II-4, R2and R3are each independently F or Cl in each formula. X1to X4are each independently N or CR x wherein three or more of X1to X4cannot be N at the same time, and R x is -H or F; R1is -CX a H2or -C(X a )2H, wherein Xa is F or Cl; and 6. A compound having a structure represented by the following formula, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:.

7. The compound according to claim 6, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is a compound represented by the following formula:.

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

9. The pharmaceutical composition according to claim 8, wherein the pharmaceutical composition is for preventing or treating a histone deacetylase 6-mediated disease. X2is N or CR x wherein R x is -H, F, Cl, Br or I; R1is -CX a H2, -C(X a )2H or -C(X a )3, wherein Xa is F, Cl, Br or I; and 10. The pharmaceutical composition according to claim 9, wherein the histone deacetylase 6-mediated disease is: an infectious disease; a neoplasm; an endocrine disease, nutritional and metabolic diseases; a mental and behavioral disorder; a neurological disease; an eye and ocular adnexa disease; a circulatory system disease; a respiratory system disease; a digestive problem; a skin and subcutaneous tissue disease; a musculoskeletal system and connective tissue disease; or a malformation, deformation, and chromosomal aberration.

11. The pharmaceutical composition according to claim 10, wherein: the infectious disease is a prion disease; the neoplasm is a benign tumor or a malignant tumor; the endocrine disease, nutritional and metabolic diseases is Wilson disease, amyloidosis, or diabetes; the mental and behavioral disorder is depression or Rett syndrome; the neurological disease is a nervous system atrophy including central nervous system atrophy, a neurodegenerative disease, a movement disorder, a neuropathy, a motor neuron disease, or a central nervous system demyelinating disease; the eye and ocular adnexa disease is uveitis; the circulatory system disease is atrial fibrillation or stroke; the respiratory system disease is asthma; ​ ​ ​ ​ ​ X2is independently at each occurrence N or CR x , and R x is H, F or Cl, Br or I; R1is independently in each formula -CX a H2, -C(X a )2H or -C(X a )3, wherein Xais independently in each formula H, F, Cl, Br or I; and ​ ​ ​ X2is independently at each occurrence N or CR x wherein R x is -H or F; R1is independently at each occurrence -C(X a )2H or -C(X a )3, wherein Xais independently at each occurrence F or Cl; and R1is independently at each occurrence -C(X a )2H or -C(X a )3, wherein Xais independently at each occurrence F or Cl; and ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ the skin and subcutaneous tissue disease is psoriasis; the musculoskeletal system and connective tissue disease is rheumatoid arthritis, osteoarthritis, or systemic lupus erythematosus; and the malformation, deformation, and chromosomal aberration is autosomal dominant polycystic kidney disease.

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

13. A pharmaceutical composition comprising the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7 as an effective ingredient for preventing or treating nervous system atrophy including central nervous system atrophy, neurodegenerative disease, or neuropathy.

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

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

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

17. Use of the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7 for the manufacture of a medicament for preventing or treating a histone deacetylase 6-mediated disease. ​

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