Novel saturated heterocycle-substituted heteroaryl-carbonohydrazonoyl dicyanide compound and use thereof

Abstract

The present invention relates to a novel saturated heterocycle-substituted heteroaryl-carbonohydrazonoyl dicyanide compound or a pharmaceutically acceptable salt thereof, a preparation method therefor, and a pharmaceutical composition comprising same as an active ingredient for preventing or treating neurological diseases. The saturated heterocycle-substituted heteroaryl-carbonohydrazonoyl dicyanide compound of the present invention inhibits the aggregation or hyperphosphorylation of tau protein and / or TDP-43, and thus can be used for preventing and treating neurological diseases including tauopathy and TDP-43 proteinopathy.

Description

Novel saturated heterocyclic substituted heteroaryl-carbonohydrazonoyl dicyanide compounds and uses thereof

[0001] The present invention relates to a novel saturated heterocyclic substituted heteroaryl-carbonohydrazonoyl dicyanide compound and its use.

[0002] Tau protein is a microtubule-associated protein (MAP) primarily expressed in the axons of neurons. It stabilizes microtubules and exhibits molecular diversity through phosphorylation. In humans, six isoforms of tau are generated by alternative splicing of mRNA, which involves insertions of 29 or 58 amino acid residues at the N-terminus and three or four repeats (called microtubule-binding domains) at the C-terminus.

[0003] In healthy neurons, tau stabilizes microtubules by promoting axonal growth and neuronal polarization. Pathologically, when hyperphosphorylated, tau detaches from microtubules, accumulating and aggregating within the cytoplasm. Hyperphosphorylation and / or aggregation of tau proteins leads to abnormal accumulation of these tau proteins in neurons, which has been implicated as a cause of various neurodegenerative diseases. Abnormal tau protein aggregation also occurs in tauopathies. While the precise role of this tau protein aggregation in the progression of tauopathies is not yet known, it appears to be similar to aggregation commonly observed in other neurodegenerative brain diseases.

[0004] The TDP-43 protein, primarily located in the nucleus, is known to be involved in gene expression by binding to DNA or RNA, inhibiting gene transcription, and regulating RNA splicing and translation. Under pathological conditions, mutations in the TDP-43 protein within the nucleus can cause it to migrate to the cytoplasm and form abnormal aggregates.

[0005] Thus, it is known that tau protein and / or Tdp-43 protein, alone or in combination, are hyperphosphorylated and / or aggregated in neurons to cause various degenerative brain diseases including tauopathies such as amyotrophic lateral sclerosis, frontotemporal dementia, and Alzheimer's disease. In particular, it has been reported that when tau protein and Tdp-43 protein aggregate in combination to cause tauopathy, the progression and severity of tau pathology are accelerated and aggravated. However, the specific mechanisms by which diseases mediated by these tau proteins and / or Tdp-43 proteins transmit signals and exhibit toxicity have not yet been elucidated, and no clear treatment or therapeutic agent is yet known to treat these diseases. Therefore, the present inventors have completed and provided an invention that can be applied to tau protein and / or Tdp-43 protein-related proteinopathies. Prior art related to the present invention includes Mol. Neurodegeneration, 2009, 4: 13, Biophys. J., 2006, 90: 3739-3748, J. Biol. Chem., 1995, 270: 7679-7688, J. Neuril. Neurosurg. Psychiatry, 2021, 92: 86-95., Front. Neuroscience, 2022, 16, 815765, Geroscience 2021, 43, 1627-1634, and Brain 2022, 145, 2769-2784.

[0006] The technical problem to be achieved by the present invention is to provide a novel saturated heterocyclic substituted heteroaryl-carbonohydrazonoyl dicyanide compound or a pharmaceutically acceptable salt thereof.

[0007] The present invention provides a method for producing the heteroaryl-carbonohydrazonoyl dicyanide compound or a pharmaceutically acceptable salt thereof.

[0008] In addition, another object of the present invention is to provide a pharmaceutical composition for preventing or treating a disease caused by aggregation or hyperphosphorylation of tau protein or Tdp-43 protein, comprising the heteroaryl-carbonohydrazonoyl dicyanide compound or a pharmaceutically acceptable salt thereof as an active ingredient.

[0009] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below.

[0010] To solve the above problem, the present invention provides a carbonohydrazonoyl dicyanide compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof:

[0011] [Chemical Formula 1]

[0012]

[0013] In the above chemical formula 1,

[0014] n is 0 or 1;

[0015] R1 is hydrogen, C 1-4 Alkyl, or C 1-4 Any one selected from alkylcarbonyl;

[0016] R2 is absent or halogen;

[0017] X is any one of O, S or NR3;

[0018] R3 is hydrogen, or C 1-4 It is alkyl;

[0019] Y1, Y2 are C or N;

[0020] Z is absent or NH;

[0021] Cy is unsubstituted or halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylcarbonyl, and C 1-4 A 5 to 10 membered saturated heterocyclyl substituted with at least one selected from the group consisting of alkoxycarbonyl.

[0022] According to one side,

[0023] R1 is any one selected from hydrogen, methyl or acetyl;

[0024] R2 is absent or chlorine;

[0025] X is any one of O, S or NR3;

[0026] R3 is any one selected from hydrogen, methyl, or ethyl;

[0027] Cy may be a 5 to 10 membered saturated heterocyclyl substituted with one or more selected from the group consisting of fluoro, methyl, isopropyl, methoxy, methylcarbonyl, and tert-butoxycarbonyl.

[0028] According to one side, the carbonohydrazonoyl dicyanide compound represented by the above [chemical formula 1] may be represented by the following [chemical formula 1-1] or [chemical formula 1-2]:

[0029] [Chemical Formula 1-1]

[0030]

[0031] [Chemical Formula 1-2]

[0032] .

[0033] According to one side,

[0034] The above compound is

[0035] 1. (2-morpholinobenzo[d]thiazol-5-yl)carbonohydrazonoyl dicyanide,

[0036] 2. (2-morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide,

[0037] 3. (2-morpholinobenzo[d]oxazol-5-yl)carbonohydrazonoyl dicyanide,

[0038] 4. (2-morpholinobenzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0039] 5. (1-methyl-2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide,

[0040] 6. (1-methyl-2-morpholino-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide,

[0041] 7. (2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide

[0042] 8. (2-(4-methylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0043] 9. (2-(4-methoxypiperidin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0044] 10. (2-(2,6-dimethylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0045] 11. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0046] 12. (2-(2-methylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0047] 13. (2-(3-methylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0048] 14. (2-(4-methylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0049] 15. (2-(2-methylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0050] 16. (2-(3-methylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0051] 17. (2-(4-methoxypiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0052] 18. (2-(2,6-dimethylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0053] 19. (5-chloro-2-morpholinobenzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0054] 20. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0055] 21. tert-butyl 4-(6-(2-(dicyanomethylene)hydrazinyl)benzo[d]oxazol-2-yl)piperazine-1-carboxylate

[0056] 22. tert-butyl 4-(6-(2-(dicyanomethylene)hydrazinyl)benzo[d]thiazol-2-yl)piperazine-1-carboxylate

[0057] 23. (2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0058] 24. (2-(piperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0059] 25. (2-(piperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0060] 26. (2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0061] 27. (2-(4-isopropylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0062] 28. (2-(4-isopropylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0063] 29. (2R,6S)-tert-butyl 4-(6-(2-(dicyanomethylene)hydrazinyl)benzo[d]thiazol-2-yl)-2,6-dimethylpiperazine-1-carboxylate

[0064] 30. (2-(4-acetylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0065] 31. (2-(4-acetylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0066] 32. (1-ethyl-2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide,

[0067] 33. (1-ethyl-2-morpholino-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide,

[0068] 34. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide)

[0069] 35. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide)

[0070] 36. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)(methyl)carbonohydrazonoyl dicyanide)

[0071] 37. Acetyl(2-morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0072] 38. (2-(morpholine-4-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0073] 39. (2-(2,6-dimethylmorpholine-4-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0074] 40. (2-(8-oxa-3-azabicyclo[3.2.1]octane-3-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0075] 41. (R)-(2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0076] 42. (S)-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0077] 43. (S)-(2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0078] 44. (2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0079] 45. (2-(pyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0080] 46. ​​(2-(piperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0081] 47.(R)-N-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0082] 48.N-(2-(morpholine-4-carbonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)carbonohydrazonoyl dicyanide, or

[0083] 49.N-(2-morpholinooxazolo[4,5-b]pyridin-6-yl)carbonohydrazonoyl dicyanide.

[0084] According to another embodiment of the present invention, a first step of forming an imine bond by reacting a compound represented by the following chemical formula 2, which includes a reactive amine group at one terminal, with sodium nitrite and malononitrile in the presence of an acid; and

[0085] Optionally, when R1 is a substituent other than hydrogen, a method for preparing a compound represented by the formula 1 is provided, which further comprises a second step of introducing an R1 substituent into the product obtained from the first step:

[0086] [Chemical Formula 1]

[0087]

[0088] [Chemical Formula 2]

[0089]

[0090] In the above chemical formulas 1 and 2,

[0091] n is 0 or 1;

[0092] R1 is hydrogen, C 1-4 Alkyl, or C 1-4 Any one selected from alkylcarbonyl;

[0093] R2 is absent or halogen;

[0094] X is any one of O, S or NR3;

[0095] R3 is hydrogen, or C 1-4 It is alkyl;

[0096] Y1, Y2 are C or N;

[0097] Z is absent or NH;

[0098] Cy is unsubstituted or halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylcarbonyl, and C 1-4 A 5 to 10 membered saturated heterocyclyl substituted with at least one selected from the group consisting of alkoxycarbonyl.

[0099] According to one side, the second stage is

[0100] i) R1 is C 1-6 In the case of alkyl, it is carried out by reacting with a halogenated alkane in the presence of potassium tert-butoxide or sodium hydride in an organic solvent,

[0101] ii) R1 is C 1-4 In the case of alkylcarbonyl, it may be carried out by reacting with a halogenated alkylcarbonyl in the presence of a base and triethylamine.

[0102] According to one side,

[0103] The compound represented by the above chemical formula 2 is

[0104] i) When n is 0 and X is S or NR3, irradiate with microwaves to react Cy-H with a compound represented by the chemical formula 3-a,

[0105] ii) When n is 0 and X is O, react Cy-H with a compound represented by chemical formula 3-b in the presence of triethylamine,

[0106] iii) When n is 1, react Cy-H with a compound represented by chemical formula 3-c,

[0107] R N In this case, the reaction product is further reduced,

[0108] iv) When Y2 is N, n is 1, and Z is NH, it can be obtained by reacting a compound represented by 3-d.

[0109] [Chemical Formula 3-a]

[0110]

[0111] [Chemical Formula 3-b]

[0112]

[0113] [Chemical Formula 3-c]

[0114]

[0115] [Chemical Formula 3-d]

[0116]

[0117] In the above chemical formulas 3-a to 3-d,

[0118] X1 is halogen, R N is nitro or amino;

[0119] R5 is any one selected from t-butoxy carbonyl, hydrogen, or benzyloxy carbonyl;

[0120] Cy is unsubstituted or halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylcarbonyl, and C 1-4 A 5 to 10 membered saturated heterocyclyl substituted with at least one selected from the group consisting of alkoxycarbonyl.

[0121] According to one side, in the compound represented by the above chemical formula 3-a, X is NR3 and R3 is C 1-4 The alkyl compound may be prepared by preparing a compound in which R3 is hydrogen by the above step i) and reacting it with halogenated R3 in the presence of DBU (1,8-Diazabicyclo[5.4.0]undec-7-ene).

[0122] According to one aspect, the compound represented by the above chemical formula 3-b may be prepared by reacting a compound represented by the following chemical formula 4 with potassium ethyl xanthogenate:

[0123] [Chemical Formula 4]

[0124] .

[0125] According to one aspect, the compound represented by the above chemical formula 3-c may be prepared by reacting the compound represented by the following chemical formula 5 with phenylchloroformate in the presence of triethylamine:

[0126] [Chemical Formula 5]

[0127] .

[0128] According to another aspect of the present invention, a pharmaceutical composition for preventing or treating a disease caused by aggregation or hyperphosphorylation of tau protein or Tdp-43 protein is provided, comprising the above-described compound as an active ingredient.

[0129] According to one aspect, the disease caused by aggregation or hyperphosphorylation of the tau protein may be selected from the group consisting of Alzheimer's disease, Parkinson's disease, vascular dementia, acute stroke, trauma, cerebrovascular disease, brain cord trauma, spinal cord trauma, peripheral neuropathy, retinopathy, glaucoma, and tauopathies.

[0130] According to one side, the above tauopathy is chronic traumatic encephalopathy (CTE), primary age-related tauopathy, progressive supranuclear palsy, corticobasal degeneration, Pick's disease, argyrophilic grain disease (AGD), frontotemporal dementia (FTD), Parkinsonism linked to chromosome 17, Lytico-Bodig disease (Parkinson-dementia complex of Guam), ganglioglioma, gangliocytoma, meningioangiomatosis, postencephalitic parkinsonism, It may be selected from the group consisting of subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, lipofuscinosis, posttraumeric stress disorder, and traumatic brain injury.

[0131] According to one side, diseases caused by aggregation or hyperphosphorylation of the TDP-43 protein include amyotrophic lateral sclerosis (ALS), familial amyotrophic lateral sclerosis due to genetic mutation (Familial ALS-SOD1), sporadic amyotrophic lateral sclerosis, superoxide dismutase 1 (SOD1), frontotemporal lobar degeneration (FTLD), amyotrophic lateral sclerosis with frontotemporal dementia (ALS-FTLD), multiple system proteinopathy (MSP), limbic-predominant age-related TDP-43 encephalopathy (LATE) / CART (cerebral age-related TDP-43 with sclerosis), Perry disease, facial onset sensory and motor neuronopathy (FOSMN), and sporadic inclusion body myositis (sIBM).

[0132] According to one aspect, the composition may inhibit the aggregation of tau protein or Tdp-43 protein.

[0133] According to one aspect, the composition may inhibit tau protein or Tdp-43 protein hyperphosphorylation.

[0134] The novel saturated heterocyclic substituted heteroaryl-carbonohydrazonoyl dicyanide compound of the present invention can effectively inhibit the aggregation and / or hyperphosphorylation of tau protein and Tdp-43, and thus can be usefully used for the prevention or treatment of various degenerative brain diseases including diseases caused thereby, such as frontotemporal dementia, Alzheimer's dementia, and tauopathy.

[0135] The effects of the present invention are not limited to the effects described above, and should be understood to include all effects that can be inferred from the detailed description of the present invention or the composition of the invention described in the claims.

[0136] Figure 1 shows the results of an experiment on the improvement of cognitive function in a tau gene-modified animal model.

[0137] Figure 2 shows the results of a BiFC fluorescence imaging experiment on the inhibition effect of tau aggregation in brain tissue.

[0138] Figure 3 shows the results of an experiment on the improvement of tau pathology through immunofluorescence staining inhibition using a tau phosphorylation antibody.

[0139] Figure 4 shows the results of an experiment to determine the efficacy of inhibiting tau pathology using brain lysate immunoblotting.

[0140] The present inventors synthesized a novel saturated heterocyclic substituted heteroaryl-carbonohydrazonoyl dicyanide compound and confirmed that it has utility for diseases related to aggregation and / or hyperphosphorylation of tau protein and Tdp-43, thereby completing the invention.

[0141] To solve the above problem, the present invention provides a carbonohydrazonoyl dicyanide compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof:

[0142] [Chemical Formula 1]

[0143]

[0144] In the above chemical formula 1,

[0145] n is 0 or 1;

[0146] R1 is hydrogen, C 1-4 Alkyl, or C 1-4 Any one selected from alkylcarbonyl;

[0147] R2 is absent or halogen;

[0148] X is any one of O, S or NR3;

[0149] R3 is hydrogen, or C 1-4 It is alkyl;

[0150] Y1, Y2 are C or N;

[0151] Z is absent or NH;

[0152] Cy is unsubstituted or halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylcarbonyl, and C 1-4 It may be, but is not limited to, a 5 to 10 membered saturated heterocyclyl substituted with one or more selected from the group consisting of alkoxycarbonyl.

[0153] Preferably,

[0154] R1 is any one selected from hydrogen, methyl or acetyl;

[0155] R2 is absent or chlorine;

[0156] X is any one of O, S or NR3;

[0157] R3 is any one selected from hydrogen, methyl, or ethyl;

[0158] Cy may be, but is not limited to, a 5- to 10-membered saturated heterocyclyl substituted with one or more selected from the group consisting of fluoro, methyl, isopropyl, methoxy, methylcarbonyl, and tert-butoxycarbonyl.

[0159] According to one side, the carbonohydrazonoyl dicyanide compound represented by the above [chemical formula 1] may be represented by the following [chemical formula 1-1] or [chemical formula 1-2]:

[0160] [Chemical Formula 1-1]

[0161]

[0162] [Chemical Formula 1-2]

[0163] .

[0164] According to one side,

[0165] The above compound is

[0166] 1. (2-morpholinobenzo[d]thiazol-5-yl)carbonohydrazonoyl dicyanide,

[0167] 2. (2-morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide,

[0168] 3. (2-morpholinobenzo[d]oxazol-5-yl)carbonohydrazonoyl dicyanide,

[0169] 4. (2-morpholinobenzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0170] 5. (1-methyl-2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide,

[0171] 6. (1-methyl-2-morpholino-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide,

[0172] 7. (2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide

[0173] 8. (2-(4-methylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0174] 9. (2-(4-methoxypiperidin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0175] 10. (2-(2,6-dimethylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0176] 11. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0177] 12. (2-(2-methylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0178] 13. (2-(3-methylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0179] 14. (2-(4-methylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0180] 15. (2-(2-methylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0181] 16. (2-(3-methylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0182] 17. (2-(4-methoxypiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0183] 18. (2-(2,6-dimethylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0184] 19. (5-chloro-2-morpholinobenzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0185] 20. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0186] 21. tert-butyl 4-(6-(2-(dicyanomethylene)hydrazinyl)benzo[d]oxazol-2-yl)piperazine-1-carboxylate

[0187] 22. tert-butyl 4-(6-(2-(dicyanomethylene)hydrazinyl)benzo[d]thiazol-2-yl)piperazine-1-carboxylate

[0188] 23. (2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0189] 24. (2-(piperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0190] 25. (2-(piperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0191] 26. (2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0192] 27. (2-(4-isopropylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0193] 28. (2-(4-isopropylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0194] 29. (2R,6S)-tert-butyl 4-(6-(2-(dicyanomethylene)hydrazinyl)benzo[d]thiazol-2-yl)-2,6-dimethylpiperazine-1-carboxylate

[0195] 30. (2-(4-acetylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0196] 31. (2-(4-acetylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0197] 32. (1-ethyl-2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide,

[0198] 33. (1-ethyl-2-morpholino-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide,

[0199] 34. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide)

[0200] 35. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide)

[0201] 36. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)(methyl)carbonohydrazonoyl dicyanide)

[0202] 37. Acetyl(2-morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0203] 38. (2-(morpholine-4-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0204] 39. (2-(2,6-dimethylmorpholine-4-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0205] 40. (2-(8-oxa-3-azabicyclo[3.2.1]octane-3-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide)

[0206] 41. (R)-(2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0207] 42. (S)-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0208] 43. (S)-(2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0209] 44. (2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide)

[0210] 45. (2-(pyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0211] 46. ​​(2-(piperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0212] 47.(R)-N-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0213] 48.N-(2-(morpholine-4-carbonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)carbonohydrazonoyl dicyanide, or

[0214] 49. N-(2-morpholinooxazolo[4,5-b]pyridin-6-yl)carbonohydrazonoyl dicyanide) may be used, and these may be represented as shown in Table 1 below.

[0215] 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49

[0216]

[0217]

[0218]

[0219] In the present invention, the term “substitution” refers to a reaction in which an atom or atomic group included in a molecule of a compound is replaced with another atom or atomic group.

[0220] In the present invention, the term "cyclo or cyclic" refers to a structure in which both ends of an organic compound skeleton are connected to form a ring. The inventors of the present invention have confirmed that when the heteroaryl side terminal in the heteroaryl-carbonohydrazonoyl-dicyanide structure is substituted with a saturated heterocyclyl, there is an excellent effect in inhibiting the aggregation and hyperphosphorylation of tau protein and / or Tdp-43.

[0221] In the present invention, the term “chain or cyclic alkyl group” means a monovalent linear or branched or cyclic saturated hydrocarbon residue consisting solely of carbon and hydrogen atoms, having 1 to 20 carbon atoms. Examples of such alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, 2-butyl, 3-butyl, pentyl, n-hexyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0222] In the present invention, the term "heterocycloalkyl group" typically refers to a saturated or unsaturated (but not aromatic) cyclohydrocarbon, which may be optionally unsubstituted, monosubstituted or polysubstituted, and in the structure of which at least one is selected from a heteroatom of N, O or S.

[0223] In the present invention, the term "aryl group" means an unsaturated aromatic ring compound having 3 to 12 carbon atoms, having a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl). Examples of such aryl groups include, but are not limited to, phenyl, naphthyl, and the like.

[0224] In the present invention, the term “heteroaryl group” refers to a single ring or multiple condensed rings in which at least one of the atoms constituting the ring has a heteroatom of N, O, or S. Examples of such heteroaryl groups include, but are not limited to, a pyridyl group, a pyrimidinyl group, a pyrazinyl group, an oxazolyl group, a furyl group, a thiophenyl group, and the like.

[0225] In the present invention, “halogen group” may be fluorine (F), chloride (Cl), bromine (Br), or iodine (I), and in the present invention, “carbonyl group” means a -C(=O)- group.

[0226] Meanwhile, the compound of the present invention may exist in the form of a pharmaceutically acceptable salt. Useful salts include acid salts formed with pharmaceutically acceptable free acids. The term "pharmaceutically acceptable salt" as used herein refers to any organic or inorganic addition salt of the compound represented by Chemical Formulae 1 to 1-2, which has a relatively non-toxic and harmless concentration that exhibits an effective effect, and whose side effects do not diminish the beneficial effects of the compound represented by Chemical Formulae 1 to 1-2.

[0227] Acid addition salts are prepared by conventional methods, for example, by dissolving the compound in an excess of an aqueous acid solution and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Equimolar amounts of the compound and an acid or alcohol (e.g., glycol monomethyl ether) in water can be heated, and the mixture can then be evaporated to dryness, or the precipitated salt can be filtered off with suction.

[0228] At this time, organic acids and inorganic acids can be used as free acids, and inorganic acids such as hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, and tartaric acid can be used, and organic acids such as methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, and hydroiodic acid can be used, but are not limited thereto.

[0229] Additionally, pharmaceutically acceptable metal salts can be prepared using bases. Alkali metal salts or alkaline earth metal salts are obtained, for example, by dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the undissolved compound salt, and evaporating and drying the filtrate. In this case, sodium, potassium, or calcium salts are particularly suitable for pharmaceutical purposes, but are not limited thereto. Furthermore, the corresponding silver salt can be prepared by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0230] Pharmaceutically acceptable salts of the compounds of the present invention include salts of acidic or basic groups that may be present in the compounds of Formulae 1 to 1-2, unless otherwise indicated. For example, pharmaceutically acceptable salts may include sodium, calcium, and potassium salts of hydroxyl groups, and other pharmaceutically acceptable salts of amino groups include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate), and p-toluenesulfonate (tosylate) salts, and may be prepared by methods for preparing salts known in the art.

[0231] As the salt of the compound of formulae 1 to 1-2 of the present invention, any salt that is pharmaceutically acceptable and exhibits pharmacological activity equivalent to that of the compound of formulae 1 to 1-2, for example, a salt of the compound of formulae 1 to 1-2 that inhibits aggregation and / or hyperphosphorylation of tau protein or Tdp-43 protein, may be used without limitation.

[0232] In addition, the compounds represented by the chemical formulae 1 to 1-2 according to the present invention include, without limitation, not only pharmaceutically acceptable salts thereof, but also solvates such as possible hydrates that can be prepared therefrom, and all possible stereoisomers. The solvates and stereoisomers of the compounds represented by the chemical formulae 1 to 1-2 can be prepared from the compounds represented by the chemical formulae 1 to 1-2 using methods known in the art.

[0233] Furthermore, the compounds represented by the chemical formulae 1 to 1-2 according to the present invention can be prepared in crystalline or amorphous form, and when prepared in crystalline form, can be optionally hydrated or solvated. The present invention may include not only stoichiometric hydrates of the compounds represented by the chemical formulae 1 to 1-2 but also compounds containing various amounts of water. The solvates of the compounds represented by the chemical formulae 1 to 1-2 according to the present invention include both stoichiometric solvates and non-stoichiometric solvates.

[0234]

[0235] According to another embodiment of the present invention, a first step of forming an imine bond by reacting a compound represented by the following chemical formula 2, which includes a reactive amine group at one terminal, with sodium nitrite and malononitrile in the presence of an acid; and

[0236] Optionally, when R1 is a substituent other than hydrogen, a method for preparing a compound represented by the formula 1 is provided, which further comprises a second step of introducing an R1 substituent into the product obtained from the first step:

[0237] [Chemical Formula 1]

[0238]

[0239] [Chemical Formula 2]

[0240]

[0241] In the above chemical formulas 1 and 2,

[0242] n is 0 or 1;

[0243] R1 is hydrogen, C 1-4 Alkyl, or C 1-4 Any one selected from alkylcarbonyl;

[0244] R2 is absent or halogen;

[0245] X is any one of O, S or NR3;

[0246] R3 is hydrogen, or C 1-4 It is alkyl;

[0247] Y1, Y2 are C or N;

[0248] Z is absent or NH;

[0249] Cy is unsubstituted or halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylcarbonyl, and C 1-4 A 5 to 10 membered saturated heterocyclyl substituted with at least one selected from the group consisting of alkoxycarbonyl.

[0250] Specifically, the first step of the above process is

[0251] Step 1-1: Dissolving the compound of chemical formula 2 and sodium nitrite in a water-containing solvent, and adding an acidic aqueous solution at -5 to 5°C to form a diazonium salt;

[0252] Step 1-2, adding malononitrile to the reaction solution containing the diazonium salt obtained from Step 1-1 and reacting at 15 to 40°C, and

[0253] It can be carried out by a series of processes including steps 1-3 of adding a base aqueous solution to the reaction solution of steps 1-2 to neutralize it, but is not limited thereto.

[0254] In addition, the first step may be performed by performing the reaction of step 1-1 using a 1 M hydrochloric acid solution in an aqueous solution at a low temperature of about 0°C while stirring for 2 minutes to 1 hour, and then performing the reaction of step 1-2 by stirring for 2 minutes to 1 hour at room temperature, but is not limited thereto.

[0255] In addition, the second step is, i) R1 is C 1-6 In the case of alkyl, it is carried out by reacting with a halogenated alkane in the presence of potassium tert-butoxide or sodium hydride in an organic solvent, and ii) R1 is C 1-4In the case of alkylcarbonyl, it can be carried out by reacting with a halogenated alkylcarbonyl in the presence of a base and triethylamine, but is not limited thereto.

[0256] Specifically, R1 is C 1-6 In the case of alkyl, the carbonohydrazonoyl dicyanide compound in which R1 is not substituted obtained from the previous step is dissolved in an organic solvent such as DMF, and a haloalkane corresponding to R1, such as an iodinated alkane, is added to the reaction solution to react at 50 to 70°C. The reaction solution may further include potassium tert-butoxide, but is not limited thereto.

[0257] Meanwhile, R1 is C 1-6 In the case of alkylcarbonyl, the carbonohydrazonoyl dicyanide compound in which R1 is not substituted obtained from the previous step is dissolved in a lower alcohol such as methanol, reacted with a base such as potassium hydroxide, and then solidified to obtain a product, which is reacted in an organic solvent such as acetonitrile in the presence of triethylamine to obtain C 1-6 The reaction can be carried out by reacting a halogenated alkylcarbonyl corresponding to the alkylcarbonyl, for example, acetyl chloride, but is not limited thereto.

[0258] According to one side,

[0259] The compound represented by the above chemical formula 2 is

[0260] i) When n is 0 and X is S or NR3, irradiate with microwaves to react Cy-H with a compound represented by the chemical formula 3-a,

[0261] ii) When n is 0 and X is O, react Cy-H with a compound represented by chemical formula 3-b in the presence of triethylamine,

[0262] iii) When n is 1, react Cy-H with a compound represented by chemical formula 3-c,

[0263] R NIn this case, the reaction product is further reduced,

[0264] iv) When Y2 is N, n is 1, and Z is NH, it can be obtained by reacting a compound represented by 3-d:

[0265] [Chemical Formula 3-a]

[0266]

[0267] [Chemical Formula 3-b]

[0268]

[0269] [Chemical Formula 3-c]

[0270]

[0271] [Chemical Formula 3-d]

[0272]

[0273] In the above chemical formulas 3-a to 3-d,

[0274] X1 is halogen, R N is nitro or amino,

[0275] R5 is any one selected from t-butoxy carbonyl, hydrogen, or benzyloxy carbonyl;

[0276] Cy is unsubstituted or halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylcarbonyl, and C 1-4 A 5 to 10 membered saturated heterocyclyl substituted with at least one selected from the group consisting of alkoxycarbonyl.

[0277] Preferably, the reduction may be carried out in an organic solvent such as 1,4-dioxane or methanol under a Pd / C catalyst, or by reaction with AcOH in the presence of Fe, or by reaction with ammonium chloride in the presence of Fe, but is not limited thereto.

[0278]

[0279] According to one side, in the compound represented by the above chemical formula 3-a, X is NR3 and R3 is C 1-4 The alkyl compound may be prepared by preparing a compound in which R3 is hydrogen by the above step i) and reacting it with halogenated R3 in the presence of DBU (1,8-Diazabicyclo[5.4.0]undec-7-ene), and preferably, this may be performed by irradiating with microwaves, but is not limited thereto.

[0280] According to one side, the compound represented by the above chemical formula 3-b may be prepared by reacting the compound represented by the following chemical formula 4 with potassium ethyl xanthogenate, and preferably, the reaction may be performed at 100 to 125°C while dissolved in an organic solvent such as pyridine, but is not limited thereto:

[0281] [Chemical Formula 4]

[0282] .

[0283] According to one side, the compound represented by the above chemical formula 3-c may be prepared by reacting the compound represented by the following chemical formula 5 with phenylchloroformate in the presence of triethylamine, and preferably, the reaction may be performed in a solvent such as dichloromethane or dimethylformamide alone or in a mixed solvent, but is not limited thereto:

[0284] [Chemical Formula 5]

[0285] .

[0286]

[0287]

[0288] In the manufacturing method of the present invention, the reactants and / or precursors used in each step may be purchased as commercially available compounds, or the reactants or intermediates of each step may be purchased and synthesized through a single reaction known in the art or by combining several reactions, but are not limited thereto.

[0289] Additionally, after each reaction, a process for isolating and / or purifying the product may be additionally included, as needed, and this can be performed using various methods known in the art.

[0290]

[0291] According to another aspect of the present invention, a pharmaceutical composition for preventing or treating a disease caused by aggregation or hyperphosphorylation of tau protein or Tdp-43 protein is provided, comprising the above-described compound as an active ingredient.

[0292] In the present invention, the term "prevention" means any act of inhibiting or delaying the occurrence, spread or recurrence of a neurological disease by administering the composition of the present invention, and "treatment" means any act of improving or beneficially changing the symptoms of the disease by administering the composition of the present invention.

[0293] The term "pharmaceutical composition" in the present invention refers to a composition manufactured for the purpose of preventing or treating a disease, and may be formulated and used in various forms according to conventional methods. For example, it may be formulated in oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, and may be formulated and used in the form of topical preparations, suppositories, and sterile injectable solutions.

[0294] In the present invention, "including as an active ingredient" means that the ingredient is included in an amount necessary or sufficient to realize a desired biological effect. In actual application, the amount included as an active ingredient can be determined by considering the amount for treating the target disease and not causing other toxicity, and may vary depending on various factors such as the disease or condition being treated, the form of the composition being administered, the size of the subject, or the severity of the disease or condition. A person of ordinary skill in the art to which the present invention pertains can empirically determine the effective amount of an individual composition without undue experimentation.

[0295] In the term "tau protein" of the present invention, tau is an abbreviation for 'tubulin associated unit', and the tau protein is one of the well-known microtubule-binding proteins, and is a group of six highly soluble protein isoforms generated by alternative splicing from the MAPT (microtubule-associated protein tau) gene. It mainly plays a role in maintaining the stability of microtubules in axons, and is abundant in neurons of the central nervous system (CNS), of which the cerebral cortex is the most abundant. Although less common elsewhere, it is also expressed at very low levels in CNS astrocytes and oligodendrocytes.

[0296] The term "TDP-43 protein" of the present invention refers to a transcriptional repressor that binds to chromosomally integrated TAR DNA and inhibits HIV-1 transcription, also known as 'transactive response DNA binding protein 43 kDa' or 'TAR DNA-binding protein 43'. In addition, the protein regulates alternative splicing of the CFTR gene. The TDP-43 protein binds to both DNA and RNA and has various functions in transcriptional repression, pre-mRNA splicing, and translational regulation.

[0297] As described above, the compound of the present invention not only inhibits the aggregation or hyperphosphorylation of tau protein and / or Tdp-43 protein but also does not exhibit toxicity to cells, so a pharmaceutical composition containing it as an active ingredient can be used for the prevention or treatment of diseases caused by the aggregation or hyperphosphorylation of tau protein or Tdp-43 protein.

[0298] For example, diseases caused by aggregation or hyperphosphorylation of tau protein to which the pharmaceutical composition of the present invention can be applied may be Alzheimer's disease, Parkinson's disease, vascular dementia, acute stroke, trauma, cerebrovascular disease, brain cord trauma, spinal cord trauma, peripheral neuropathy, retinopathy, glaucoma, or tauopathies.Non-limiting examples of the above tauopathies include chronic traumatic encephalopathy (CTE), primary age-related tauopathy, progressive supranuclear palsy, corticobasal degeneration, Pick's disease, argyrophilic grain disease (AGD), frontotemporal dementia (FTD), Parkinsonism linked to chromosome 17, Lytico-Bodig disease (Parkinson-dementia complex of Guam), ganglioglioma, gangliocytoma, meningioangiomatosis, postencephalitic parkinsonism, These include subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, lipofuscinosis, posttraumatic stress disorder, and traumatic brain injury.

[0299] In addition, diseases caused by aggregation or hyperphosphorylation of TDP-43 protein to which the pharmaceutical composition of the present invention can be applied include amyotrophic lateral sclerosis (ALS), familial amyotrophic lateral sclerosis due to genetic mutation (Familial ALS-SOD1), sporadic amyotrophic lateral sclerosis, superoxide dismutase 1 (SOD1), frontotemporal lobar degeneration (FTLD), amyotrophic lateral sclerosis with frontotemporal dementia (ALS-FTLD), multiple system proteinopathy (MSP), limbic-predominant age-related TDP-43 encephalopathy (LATE) / CART (cerebral These include age-related TDP-43 with sclerosis, Perry disease, facial onset sensory and motor neuronopathy (FOSMN), and sporadic inclusion body myositis (sIBM).

[0300] In addition, the pharmaceutical composition of the present invention may, depending on each formulation, additionally include one or more pharmaceutically acceptable carriers in addition to the above-described effective ingredients.

[0301] The pharmaceutically acceptable carrier may be saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture of one or more of these components, and may further include other conventional additives such as antioxidants, buffers, and bacteriostatic agents, if necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into an injectable formulation such as an aqueous solution, suspension, or emulsion, or into a pill, capsule, granule, or tablet. Furthermore, the composition may be preferably formulated according to each disease or component using an appropriate method in the art or a method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA).

[0302] The composition of the present invention can be administered orally or parenterally in a pharmaceutically effective amount depending on the intended method, and the term “pharmaceutically effective amount” of the present invention means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects, and the effective dosage level can be determined based on factors including the patient’s health condition, severity, activity of the drug, sensitivity to the drug, administration method, administration time, administration route and excretion rate, treatment period, drugs used in combination or simultaneously, and other factors well known in the medical field.

[0303] Therefore, by administering the pharmaceutical composition of the present invention to a subject, diseases associated with aggregation or hyperphosphorylation of tau protein or Tdp-43 can be prevented, treated, and / or diagnosed, examples of which are as described above.

[0304] In the present invention, the term “subject” is not limited to a mammal such as a livestock or a human that requires prevention, treatment, and / or diagnosis of the above-described exemplary disease, but may preferably be a human.

[0305] The pharmaceutical composition of the present invention can be formulated in various forms for administration to a subject, and a representative form for parenteral administration is an injectable form, preferably an isotonic aqueous solution or suspension.

[0306] The above formulations for parenteral administration can be prepared by conventional mixing, granulating, or coating methods and can contain the active ingredient in a range of about 0.1 to 75 wt%, preferably about 1 to 50 wt%. A unit dosage form for a mammal weighing about 50 to 70 kg can contain about 10 to 200 mg of the active ingredient. Injectable formulations can be prepared according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. For example, each component can be dissolved in saline or a buffer to prepare an injectable formulation. In addition, oral administration dosage forms include, for example, ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers, which may contain, in addition to the active ingredient, diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine) and lubricants (e.g., silica, talc, stearic acid and its magnesium or calcium salts, and / or polyethylene glycol). The tablets may contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidine, and may optionally further contain disintegrating agents such as starch, agar, alginic acid or its sodium salt, absorbents, coloring agents, flavoring agents, and / or sweetening agents. The above formulation can be prepared by conventional mixing, granulating or coating methods.

[0307] In addition, the pharmaceutical composition of the present invention may further include auxiliary agents such as preservatives, wetting agents, emulsifying agents, salts for osmotic pressure control or buffers, and other therapeutically useful substances, and may be formulated according to conventional methods.

[0308] The pharmaceutical composition according to the present invention can be administered via various routes, including oral, transdermal, subcutaneous, intravenous, or intramuscular. The dosage of the active ingredient can be appropriately selected based on various factors, such as the route of administration, the patient's age, sex, weight, and severity of the condition. Furthermore, the composition of the present invention can be administered in combination with known compounds capable of enhancing the desired effect.

[0309] The pharmaceutical composition according to the present invention may be administered to humans or animals orally or parenterally, such as intravenously, subcutaneously, intranasally, or intraperitoneally. Oral administration also includes sublingual administration. Parenteral administration includes injection methods such as subcutaneous injection, intramuscular injection, and intravenous injection, as well as drip methods.

[0310] In the pharmaceutical composition of the present invention, the total effective amount of the heteroaryl-carbonohydrazonoyl-dicyanide compound or a pharmaceutically acceptable salt thereof according to the present invention can be administered to a patient as a single dose, or can be administered by a fractionated treatment protocol in which multiple doses are administered over a long period of time. The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the severity of the disease, but can typically be administered repeatedly several times a day at an effective dose of 100 μg to 3,000 mg per administration for an adult. However, the concentration of the heteroaryl-carbonohydrazonoyl-dicyanide compound or a pharmaceutically acceptable salt thereof can be determined by taking into consideration various factors such as the route of administration and the number of treatments as well as the patient's age, weight, health status, sex, severity of the disease, diet, and excretion rate.

[0311] In addition, the pharmaceutical composition according to the present invention is not particularly limited in its formulation, administration route, and administration method as long as it exhibits the effects of the present invention, and the pharmaceutical composition of the present invention may additionally include a known drug in addition to the heteroaryl-carbonohydrazonoyl dicyanide compound or a pharmaceutically acceptable salt thereof as an active ingredient, and may be used in combination with other known treatments for the treatment of these diseases.

[0312] According to one aspect, the composition may inhibit the aggregation of tau protein or Tdp-43 protein.

[0313] According to one aspect, the composition may inhibit tau protein or Tdp-43 protein hyperphosphorylation.

[0314] As another aspect of the present invention, a method for preventing or treating a disease caused by aggregation or hyperphosphorylation of tau protein or Tdp-43 protein is provided, comprising administering the pharmaceutical composition of the present invention to a subject in need thereof.

[0315] By administering the pharmaceutical composition of the present invention to a subject, the above-mentioned disease can be effectively prevented or treated. Furthermore, since the pharmaceutical composition of the present invention exhibits a therapeutic effect by inhibiting the aggregation or hyperphosphorylation of tau protein or Tdp-43 protein, it can exhibit a synergistic effect when administered in combination with existing therapeutic agents.

[0316] The term "administration" in the present invention means providing a predetermined substance to a patient by any suitable method, and the route of administration of the composition of the present invention may be administered through any common route as long as it can reach the target tissue. It may be intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, or rectal administration, but is not limited thereto. In addition, the pharmaceutical composition of the present invention may be administered by any device that allows the active substance to travel to the target cell. Preferred administration methods and formulations include intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection, and drip injection. Injectables can be manufactured using aqueous solvents such as saline solution and Ringer's solution, non-aqueous solvents such as vegetable oils, higher fatty acid esters (e.g., ethyl oleate, etc.), alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, etc.), and pharmaceutical carriers such as stabilizers to prevent deterioration (e.g., ascorbic acid, sodium bisulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers to adjust pH, and preservatives to inhibit microbial growth (e.g., phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

[0317]

[0318] The terms used in the examples are for illustrative purposes only and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as "comprise" or "have" are intended to specify the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but should be understood to not preclude the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0319] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiments pertain. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0320] The present invention is susceptible to various modifications and embodiments. Specific embodiments are illustrated in the drawings and described in detail in the following detailed description. However, this is not intended to limit the present invention to specific embodiments, but rather to encompass all modifications, equivalents, and alternatives falling within the spirit and technical scope of the present invention. In describing the present invention, detailed descriptions of related known technologies will be omitted if they are deemed to obscure the gist of the present invention.

[0321] Hereinafter, the present invention will be described in more detail through examples. These examples are intended to explain the present invention more specifically, and the scope of the present invention is not limited by these examples.

[0322] Example 1: Preparation of (2-morpholinobenzo[d]thiazol-5-yl)carbonohydrazonoyl dicyanide (compound 1)

[0323] Step 1-1: Preparation of 2-morpholinobenzo[d]thiazol-5-amine

[0324]

[0325] 2-Chlorobenzo[d]thiazol-5-amine (300 mg, 1.62 mmol) was dissolved in morpholine (3 mL), and the mixture was irradiated with microwaves and reacted at 180°C for 10 minutes. Distilled water was then added, and the mixture was stirred at room temperature for 1 hour and filtered to obtain 311 mg (yield 81%) of the title compound.

[0326] 1 H NMR (400 MHz, DMSO-d6) δ7.34 (d,J= 8.4 Hz, 1H), 6.69 (d,J= 2.2 Hz, 1H), 6.43 - 6.34 (m, 1H), 5.00 (s, 2H), 3.71 (t,J= 4.9 Hz, 4H), 3.47 (t,J= 4.9 Hz, 4H).

[0327]

[0328] Step 1-2: Preparation of (2-morpholinobenzo[d]thiazol-5-yl)carbonohydrazonoyl dicyanide

[0329]

[0330] 2-Morpholinobenzo[d]thiazol-5-amine (300 mg, 1.27 mmol) and sodium nitrite (132 mg, 1.91 mmol), prepared according to the above step 1-1, were dissolved in distilled water, and then 1.0 M aqueous hydrochloric acid solution (3.8 mL, 3.82 mmol) was added at 0°C. The reaction mixture was stirred at 0°C for 10 minutes to form a diazonium salt. To the reaction mixture containing the diazonium salt, malononitrile (168 mg, 2.55 mmol) was added and stirred at room temperature for 10 minutes. Thereafter, the pH of the reaction solution was adjusted to 5.0 with aqueous sodium hydroxide and stirred at room temperature for 1 hour. Upon completion of the reaction, the produced solid was filtered to obtain 362 mg (yield 91%) of the title compound.

[0331] 1H NMR (400 MHz, DMSO-d6) δ13.03 (s, 1H), 7.80 (d,J= 8.6 Hz, 1H), 7.51 (d,J= 2.1 Hz, 1H), 7.23 (dd,J= 8.6, 2.2 Hz, 1H), 3.73 (t,J= 4.9 Hz, 4H), 3.56 (t,J= 4.8 Hz, 4H).

[0332]

[0333] Example 2: Preparation of (2-morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (compound 2)

[0334] Step 2-1: Preparation of 2-morpholinobenzo[d]thiazol-5-amine

[0335]

[0336] A reaction was performed in a similar manner to step 1-1 of Example 1, except that 2-chlorobenzo[d]thiazol-6-amine (300 mg, 1.62 mmol) was used instead of 2-chlorobenzo[d]thiazol-5-amine, to obtain 338 mg (yield 88%) of the title compound.

[0337] 1 H NMR (400 MHz, DMSO-d6) δ7.19 (d,J= 8.5 Hz, 1H), 6.91 (d,J= 2.3 Hz, 1H), 6.57 (dd,J= 8.5, 2.3 Hz, 1H), 4.92 (s, 2H), 3.74 - 3.67 (m, 4H), 3.45 - 3.38 (m, 4H).

[0338]

[0339] Step 2-2: Preparation of (2-morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0340]

[0341] Except that 2-morpholinobenzo[d]thiazol-6-amine (300 mg, 1.27 mmol) obtained from step 2-1 was used instead of 2-morpholinobenzo[d]thiazol-5-amine, a reaction was carried out in a similar manner to step 1-2 of Example 1, to obtain 219 mg (yield 55%) of the title compound.

[0342] 1 H NMR (400 MHz, DMSO-d6) δ13.10 (s, 1H), 7.92 (d,J= 2.1 Hz, 1H), 7.51 - 7.40 (m, 2H), 3.72 (t,J= 4.9 Hz, 4H), 3.55 (t,J= 4.9 Hz, 4H).

[0343]

[0344] Example 3: Preparation of (2-morpholinobenzo[d]oxazol-5-yl)carbonohydrazonoyl dicyanide (compound 3)

[0345] Step 3-1: Preparation of 5-nitrobenzo[d]oxazole-2(3H)-thione

[0346]

[0347] 2-Amino-4-nitrophenol (1 g, 6.49 mmol) was dissolved in pyridine, potassium ethyl xanthogenate (1.14 g, 7.14 mmol) was added, and the mixture was stirred at 120°C for 8 hours. 2 N HCl was added to the reaction mixture at room temperature until a solid reactant was formed, and the mixture was stirred for 30 minutes, then filtered with distilled water to obtain 528 mg (yield 41%) of the title compound.

[0348] 1 H NMR (400 MHz, DMSO-d6) δ8.19 (dq,J= 8.9, 2.3 Hz, 1H), 7.94 (d,J= 2.0 Hz, 1H), 7.73 (dd,J= 9.1, 3.3 Hz, 1H).

[0349]

[0350] Step 3-2: Preparation of 2-morpholino-5-nitrobenzo[d]oxazole

[0351]

[0352] 5-Nitrobenzo[d]oxazole-2(3H)-thione (100 mg, 0.51 mmol) prepared according to the above step 3-1 was dissolved in toluene, morpholine (53 μL, 0.61 mmol) and triethylamine (213 μL, 1.53 mmol) were added, and the mixture was stirred at 150°C for 12 hours. Upon completion of the reaction, the mixture was extracted with distilled water and ethyl acetate, and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, solidified with hexane and ether, and then filtered to obtain 45 mg (yield 35%) of the title compound.

[0353] 1 H NMR (400 MHz, DMSO-d6) δ8.09 (d,J= 2.3 Hz, 1H), 7.99 (dd,J= 8.8, 2.4 Hz, 1H), 7.66 (d,J= 8.8 Hz, 1H), 3.78 - 3.70 (m, 4H), 3.71 - 3.61 (m, 4H).

[0354]

[0355] Step 3-3: Preparation of 2-morpholinobenzo[d]oxazol-5-amine

[0356]

[0357]

[0358] 2-Morpholino-5-nitrobenzo[d]oxazole (150 mg, 0.60 mmol) and 10% Pd / C (128 mg, 0.12 mmol) prepared according to the above step 3-2 were dissolved in 1,4-dioxane and stirred at room temperature for 4 hours under hydrogen. Upon completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain 83 mg (yield 63%) of the title compound.

[0359] 1H NMR (400 MHz, DMSO-d6) δ7.03 (d,J= 8.4 Hz, 1H), 6.50 (d,J= 2.2 Hz, 1H), 6.25 (dd,J= 8.5, 2.2 Hz, 1H), 4.79 (s, 2H), 3.72 - 3.66 (m, 4H), 3.54 - 3.48 (m, 4H).

[0360]

[0361] Step 3-4: Preparation of (2-morpholinobenzo[d]oxazol-5-yl)carbonohydrazonoyl dicyanide

[0362]

[0363] A reaction was carried out in a similar manner to step 1-2 of Example 1, except that 2-morpholinobenzo[d]oxazol-5-amine (60 mg, 0.27 mmol) obtained from step 3-3 was used instead of 2-morpholinobenzo[d]thiazol-5-amine, to obtain 62 mg (yield 85%) of the title compound.

[0364] 1 H NMR (400 MHz, DMSO-d6) δ13.01 (s, 1H), 7.44 (d,J= 8.6 Hz, 1H), 7.35 (d,J= 2.2 Hz, 1H), 7.17 (dd,J= 8.6, 2.2 Hz, 1H), 3.75 - 3.68 (m, 5H), 3.63 - 3.56 (m, 3H).

[0365]

[0366] Example 4: Preparation of (2-morpholinobenzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 4)

[0367]

[0368] Step 4-1: Preparation of 6-nitrobenzo[d]oxazole-2(3H)-thione

[0369]

[0370] A reaction was performed in a similar manner to step 3-1 of Example 3, except that 2-amino-5-nitrophenol (1 g, 6.49 mmol) was used instead of 2-amino-4-nitrophenol, to obtain 373 mg (yield 29%) of the title compound.

[0371] 1 H NMR (400 MHz, DMSO-d6) δ14.37 (s, 1H), 8.43 (d,J= 2.2 Hz, 1H), 8.23 ​​(dd,J= 8.7, 2.2 Hz, 1H), 7.41 (d,J= 8.7 Hz, 1H).

[0372]

[0373] Step 4-2: Preparation of 2-morpholino-6-nitrobenzo[d]oxazole

[0374]

[0375] Except that 6-nitrobenzo[d]oxazole-2(3H)-thione (350 mg, 1.78 mmol) obtained from step 4-1 was used instead of 5-nitrobenzo[d]oxazole-2(3H)-thione, a reaction was performed in a similar manner to step 3-2 of Example 3, to obtain 220 mg (yield 49%) of the title compound.

[0376] 1 H NMR (400 MHz, DMSO-d6) δ8.31 (d,J= 2.3 Hz, 1H), 8.16 (dd,J= 8.7, 2.3 Hz, 1H), 7.42 (d,J= 8.7 Hz, 1H), 3.78 - 3.71 (m, 4H), 3.71 - 3.66 (m, 4H).

[0377]

[0378] Step 4-3: Preparation of 2-morpholinobenzo[d]oxazol-6-amine

[0379]

[0380] A reaction was carried out in a similar manner to step 3-3 of Example 3, except that 2-morpholino-6-nitrobenzo[d]oxazole (200 mg, 0.80 mmol) obtained from step 4-2 was used instead of 2-morpholino-5-nitrobenzo[d]oxazole, to obtain 130 mg (yield 74%) of the title compound.

[0381] 1 H NMR (400 MHz, DMSO-d6) δ6.98 (d,J= 8.3 Hz, 1H), 6.63 (d,J= 2.0 Hz, 1H), 6.42 (dd,J= 8.3, 2.1 Hz, 1H), 4.90 (s, 2H), 3.69 (t,J= 4.8 Hz, 4H), 3.46 (t,J= 4.8 Hz, 4H).

[0382]

[0383] Step 4-4: Preparation of (2-morpholinobenzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0384]

[0385] A reaction was carried out in a similar manner to step 1-2 of Example 1, except that 2-morpholinobenzo[d]oxazol-6-amine (100 mg, 0.46 mmol) obtained from step 4-3 was used instead of 2-morpholinobenzo[d]thiazol-5-amine, to obtain 116 mg (yield 94%) of the title compound.

[0386] 1 H NMR (400 MHz, DMSO-d6) δ13.08 (s, 1H), 7.51 (d,J= 1.8 Hz, 1H), 7.39 - 7.27 (m, 2H), 3.76 - 3.68 (m, 4H), 3.63 - 3.53 (m, 4H).

[0387]

[0388] Example 5: Preparation of (1-methyl-2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide (Compound 5)

[0389]

[0390] Step 5-1: Preparation of 4-(6-nitro-1H-benzo[d]imidazol-2-yl)morpholine

[0391]

[0392] 2-Chloro-6-nitro-1H-benzo[d]imidazole (500 mg, 2.53 mmol) was dissolved in morpholine (6 mL) and reacted at 180°C for 10 minutes under microwave irradiation. Upon completion of the reaction, the mixture was extracted with distilled water and ethyl acetate, and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, solidified with ether, and filtered to obtain 379 mg (yield 60%) of the title compound.

[0393] 1 H NMR (400 MHz, DMSO-d6) δ12.03 (s, 1H), 7.99 (d,J= 2.3 Hz, 1H), 7.94 (dd,J= 8.7, 2.3 Hz, 1H), 7.31 (d,J= 8.7 Hz, 1H), 3.79 - 3.70 (m, 4H), 3.62 - 3.51 (m, 4H).

[0394]

[0395] Step 5-2: Preparation of 4-(1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)morpholine (5-2-1) and 4-(1-methyl-5-nitro-1H-benzo[d]imidazol-2-yl)morpholine (5-2-2)

[0396]

[0397] 4-(6-Nitro-1H-benzo[d]imidazol-2-yl)morpholine (1 g, 4.03 mmol) and DBU (1,8-Diazabicyclo[5.4.0]undec-7-ene, 1.84 mL, 12.08 mmol) obtained from the above step 5-1 and methyl iodide (752 μL, 12.08 mmol) were dissolved in dimethylformamide, irradiated with microwave, and reacted at 180°C for 4 hours. Upon completion of the reaction, the mixture was extracted with distilled water and ethyl acetate, and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and separated by column chromatography to obtain 100 mg (yield 9%) of the title compounds 5-2-1 and 118 mg (yield 11%) of 5-2-2.

[0398] 5-2-1: 1 H NMR (400 MHz, DMSO-d6) δ8.33 (d,J= 2.3 Hz, 1H), 8.04 (dd,J= 8.8, 2.3 Hz, 1H), 7.51 (d,J= 8.8 Hz, 1H), 3.82 - 3.77 (m, 4H), 3.73 (s, 3H), 3.44 - 3.37 (m, 4H),

[0399] 5-2-2: 1 H NMR (400 MHz, DMSO-d6) δ8.25 (d,J= 2.2 Hz, 1H), 8.06 (dd,J= 8.8, 2.3 Hz, 1H), 7.58 (d,J= 8.8 Hz, 1H), 3.82 - 3.77 (m, 4H), 3.71 (s, 3H), 3.35 - 3.32 (m, 4H).

[0400]

[0401] Step 5-3: Preparation of 1-methyl-2-morpholino-1H-benzo[d]imidazol-6-amine

[0402]

[0403] 4-(1-Methyl-6-nitro-1H-benzo[d]imidazol-2-yl)morpholine (80 mg, 0.30 mmol) and ammonium chloride (130 mg, 2.44 mmol) obtained from the above step 5-2 were dissolved in distilled water and tetrahydrofuran and stirred at room temperature for 30 minutes. Iron (136 mg, 2.44 mmol) was added to the reaction mixture and stirred at 60°C for 3 hours. Upon completion of the reaction, the mixture was filtered through a filter paper and extracted with distilled water and ethyl acetate. The organic layer was then dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and separated by column chromatography to obtain 54 mg (yield 76%) of the title compound.

[0404] 1 H NMR (400 MHz, DMSO-d6) δ7.08 (d,J= 8.3 Hz, 1H), 6.50 - 6.38 (m, 2H), 4.78 (s, 2H), 3.76 (t,J= 4.6 Hz, 4H), 3.46 (s, 3H), 3.10 (t,J= 4.7 Hz, 4H).

[0405]

[0406] Step 5-4: Preparation of (1-methyl-2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide

[0407]

[0408]

[0409] A reaction was carried out in a similar manner to step 1-2 of Example 1, except that 1-methyl-2-morpholino-1H-benzo[d]imidazol-6-amine (50 mg, 0.21 mmol) obtained from step 5-3 was used instead of 2-morpholinobenzo[d]thiazol-5-amine, to obtain 50 mg (yield 74%) of the title compound.

[0410] 1H NMR (400 MHz, DMSO-d6) δ7.50 - 7.40 (m, 2H), 7.32 (dd,J= 8.5, 2.0 Hz, 1H), 3.79 (t,J= 4.7 Hz, 4H), 3.65 (s, 3H), 3.34 - 3.31 (m, 4H).

[0411]

[0412] Example 6: Preparation of (1-methyl-2-morpholino-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide (compound 6)

[0413]

[0414] Step 6-1: Preparation of 1-methyl-2-morpholino-1H-benzo[d]imidazol-5-amine

[0415]

[0416] Except that 4-(1-methyl-5-nitro-1H-benzo[d]imidazol-2-yl)morpholine (40 mg, 0.17 mmol) obtained from step 5-2 of Example 5 was used instead of 4-(1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)morpholine, the reaction was carried out in the same manner as step 5-3 of Example 5, to obtain 67 mg (yield 75%) of the title compound.

[0417] 1 H NMR (400 MHz, DMSO-d6) δ6.99 (d,J= 8.3 Hz, 1H), 6.62 (d,J= 2.1 Hz, 1H), 6.43 (dd,J= 8.3, 2.1 Hz, 1H), 4.58 (s, 2H), 3.76 (t,J= 4.7 Hz, 4H), 3.49 (s, 3H), 3.14 (t,J= 4.7 Hz, 4H).

[0418]

[0419] Step 6-2: Preparation of (1-methyl-2-morpholino-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide

[0420]

[0421] Except that 1-methyl-2-morpholino-1H-benzo[d]imidazol-5-amine (50 mg, 0.21 mmol) obtained from step 6-1 was used instead of 2-morpholinobenzo[d]thiazol-5-amine, the reaction was carried out in a similar manner to step 1-2 of Example 1, to obtain 56 mg (yield 88%) of the title compound.

[0422] 1 H NMR (400 MHz, DMSO-d6) δ13.08 (s, 1H), 7.52 - 7.37 (m, 2H), 7.30 (dd,J= 8.7, 2.0 Hz, 1H), 3.78 (t,J= 4.6 Hz, 4H), 3.63 (s, 3H).

[0423]

[0424] Example 7: Preparation of (2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide (compound 7)

[0425]

[0426] Step 7-1: Preparation of 1-methyl-2-morpholino-1H-benzo[d]imidazol-6-amine

[0427]

[0428] Except that 4-(1-methyl-5-nitro-1H-benzo[d]imidazol-2-yl)morpholine (400 mg, 1.61 mmol) obtained from step 5-2 of Example 5 was used instead of 4-(1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)morpholine, the reaction was carried out in the same manner as step 5-3 of Example 5, to obtain 66 mg (yield 19%) of the title compound.

[0429] 1H NMR (400 MHz, DMSO-d6) δ10.94 (s, 1H), 6.88 (d,J= 8.2 Hz, 1H), 6.47 (d,J= 2.0 Hz, 1H), 6.27 (dd,J= 8.2, 2.1 Hz, 1H), 4.50 (s, 2H), 3.70 (t,J= 4.8 Hz, 4H), 3.35 (t,J= 4.8 Hz, 4H).

[0430]

[0431] Step 7-2: Preparation of (2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide

[0432]

[0433] Except that 1-methyl-2-morpholino-1H-benzo[d]imidazol-6-amine (60 mg, 0.27 mmol) obtained from step 7-1 was used instead of 2-morpholinobenzo[d]thiazol-5-amine, the reaction was carried out in a similar manner to step 1-2 of Example 1, to obtain 38 mg (yield 47%) of the title compound.

[0434] 1 H NMR (400 MHz, DMSO-d6) δ12.57 (s, 1H), 7.31 - 7.23 (m, 2H), 7.20 (dd,J= 8.5, 1.9 Hz, 1H), 3.77 (t,J= 4.8 Hz, 4H), 3.54 (t,J= 4.8 Hz, 4H).

[0435]

[0436] Example 8: Preparation of (2-(4-methylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (Compound 8)

[0437]

[0438] Step 8-1: Preparation of 2-(4-methylpiperazin-1-yl)benzo[d]thiazol-6-amine

[0439]

[0440] 2-Chlorobenzo[d]thiazol-6-amine (300 mg, 1.62 mmol) was dissolved in 1-methylpiperazine (2.2 mL, 19.50 mmol) and reacted at 180°C for 10 minutes under microwave irradiation. Distilled water was then added, the mixture was stirred at room temperature for 1 hour, extracted with ethyl acetate, and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, solidified with ether, and filtered to obtain 179 mg (yield 44%) of the title compound.

[0441] 1 H NMR (400 MHz, DMSO-d6) δ7.16 (d,J= 8.5 Hz, 1H), 6.89 (d,J= 2.3 Hz, 1H), 6.56 (dd,J= 8.5, 2.3 Hz, 1H), 4.89 (s, 2H), 3.43 (t,J= 5.1 Hz, 4H), 3.32 (s, 4H), 2.41 (t,J= 5.1 Hz, 4H).

[0442]

[0443] Step 8-2: Preparation of (2-(4-methylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0444]

[0445] A reaction was carried out in a similar manner to step 1-2 of Example 1, except that 2-(4-methylpiperazin-1-yl)benzo[d]thiazol-6-amine (100 mg, 0.40 mmol) obtained from step 8-1 was used instead of 2-morpholinobenzo[d]thiazol-5-amine, to obtain 103 mg (yield 78%) of the title compound.

[0446] 1H NMR (400 MHz, DMSO-d6) δ10.46 (s, 1H), 7.74 (d,J= 2.1 Hz, 1H), 7.41 (d,J= 8.6 Hz, 1H), 7.30 (dd,J= 8.6, 2.1 Hz, 1H), 3.77 (s, 4H), 3.24 (s, 4H), 2.77 (s, 3H).

[0447]

[0448] Example 9: Preparation of (2-(4-methoxypiperidin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (Compound 9)

[0449]

[0450] Step 9-1: Preparation of 2-(4-methoxypiperidin-1-yl)benzo[d]thiazol-6-amine

[0451]

[0452] 2-Chlorobenzo[d]thiazol-6-amine (200 mg, 1.08 mmol) was dissolved in toluene, 4-methoxypiperidine (1.34 mL, 10.83 mmol) was added, and the mixture was irradiated with microwave and reacted at 180°C for 30 minutes. Distilled water was added, the mixture was stirred at room temperature for 1 hour, extracted with ethyl acetate, and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and purified by column chromatography to obtain 270 mg (yield 95%) of the title compound.

[0453] 1 H NMR (400 MHz, DMSO-d6) δ7.15 (d,J= 8.5 Hz, 1H), 6.88 (d,J= 2.4 Hz, 1H), 6.55 (dd,J= 8.5, 2.4 Hz, 1H), 4.88 (s, 2H), 3.76 - 3.61 (m, 2H), 3.43 (dq,J= 8.2, 4.0 Hz, 1H), 3.29 - 3.23 (m, 5H), 1.50 (qd,J= 8.9, 4.5 Hz, 2H)

[0454]

[0455] Step 9-2: Preparation of (2-(4-methoxypiperidin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0456]

[0457] Except that 2-(4-methoxypiperidin-1-yl)benzo[d]thiazol-6-amine (200 mg, 0.76 mmol) obtained from step 9-1 was used instead of 2-morpholinobenzo[d]thiazol-5-amine, the reaction was carried out in a similar manner to step 1-2 of Example 1, to obtain 34 mg (yield 13%) of the title compound.

[0458] 1 H NMR (400 MHz, DMSO-d6) δ13.09 (s, 1H), 7.91 (d,J= 2.1 Hz, 1H), 7.50 - 7.39 (m, 2H), 3.88 (d,J= 12.5 Hz, 2H), 3.69 (dt,J= 6.7, 3.7 Hz, 2H), 2.82 (dd,J= 12.7, 10.6 Hz, 2H), 1.17 (d,J= 6.2 Hz, 6H).

[0459]

[0460] Example 10: Preparation of (2-(2,6-dimethylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (Compound 10)

[0461]

[0462] Step 10-1: Preparation of 2-(2,6-dimethylmorpholino)benzo[d]thiazol-6-amine

[0463]

[0464] The reaction was carried out in a similar manner to step 9-1 of Example 9, except that 2,6-dimethylmorpholine (1.33 mL, 10.83 mmol) was used instead of 4-methoxypiperidine, to obtain 175 mg (yield 60%) of the title compound.

[0465] 1H NMR (400 MHz, DMSO-d6) δ7.17 (d,J= 8.5 Hz, 1H), 6.90 (d,J= 2.3 Hz, 1H), 6.57 (dd,J= 8.5, 2.3 Hz, 1H), 4.91 (s, 2H), 3.75 (dd,J= 12.8, 2.4 Hz, 2H), 3.71 - 3.61 (m, 2H), 2.69 (dd,J= 12.5, 10.5 Hz, 2H), 1.15 (d,J= 6.2 Hz, 6H).

[0466]

[0467] Step 10-2: Preparation of (2-(2,6-dimethylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0468]

[0469] A reaction was carried out in a similar manner to step 1-2 of Example 1, except that 2-(2,6-dimethylmorpholino)benzo[d]thiazol-6-amine (100 mg, 0.54 mmol) obtained from step 10-1 was used instead of 2-morpholinobenzo[d]thiazol-5-amine, to obtain 81 mg (yield 44%) of the title compound.

[0470] 1 H NMR (400 MHz, DMSO-d6) δ13.09 (s, 1H), 7.91 (d,J= 2.1 Hz, 1H), 7.61 - 7.24 (m, 2H), 3.88 (d,J= 12.5 Hz, 2H), 3.69 (td,J= 6.6, 3.1 Hz, 2H), 2.82 (dd,J= 12.7, 10.6 Hz, 2H), 1.17 (d,J= 6.2 Hz, 6H).

[0471]

[0472] Example 11: Preparation of (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (Compound 11)

[0473]

[0474] Step 11-1: Preparation of 2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-amine

[0475]

[0476] The reaction was carried out in a similar manner to step 9-1 of Example 9, except that 3-oxa-8-azabicyclo[3.2.1]octane (613 mg, 0.54 mmol) was used instead of 4-methoxypiperidine, to obtain 262 mg (yield 91%) of the title compound.

[0477] 1 H NMR (400 MHz, Chloroform-d) δ7.38 (d,J= 8.5 Hz, 1H), 6.96 (d,J= 2.3 Hz, 1H), 6.69 (dd,J= 8.5, 2.4 Hz, 1H), 4.22 (s, 2H), 3.95 (d,J= 11.0 Hz, 2H), 3.62 (d,J= 11.0 Hz, 4H), 2.10 (s, 4H).

[0478] Step 11-2: Preparation of N-(2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (compound 11)

[0479]

[0480]

[0481] In the same manner as in Step 1-2, 2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-amine (150 mg, 0.57 mmol), sodium nitrite (59 mg, 0.86 mmol) and 1.0 M aqueous hydrochloric acid solution (1.7 mL, 1.72 mmol) were dissolved in distilled water, and then the resulting mixture was treated with malononitrile (76 mg, 1.15 mmol) and aqueous sodium hydroxide solution to obtain 90 mg (yield 46%) of the title compound.

[0482] 1H NMR (400 MHz, DMSO-d) δ13.10 (s, 1H), 7.92 (d, J = 2.2 Hz, 1H), 7.61 - 7.32 (m, 2H), 4.28 (s, 2H), 3.75 (d, J = 11.0 Hz, 2H), 3.61 (d, J = 11.0 Hz, 2H), 2.00 (s, 4H).

[0483]

[0484]

[0485] Example 12: Preparation of N-(2-(2-methylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (Compound 12)

[0486] Step 12-1: Preparation of 2-(2-methylmorpholino)benzo[d]thiazol-6-amine (Compound 12-1)

[0487]

[0488]

[0489] In the same manner as in step 9-1, 2-chloro-6-benzothiazolamine (100 mg, 0.54 mmol) was dissolved in toluene, and then 2-methylmorpholine (0.58 mL, 5.42 mmol) was added to obtain 84 mg (yield 62%) of the title compound.

[0490] 1 H NMR (400 MHz, DMSO-d6) δ7.18 (d, J = 8.5 Hz, 1H), 6.91 (d, J = 2.3 Hz, 1H), 6.57 (dd, J = 8.5, 2.3 Hz, 1H), 4.91 (s, 2H), 3.95 - 3.86 (m, 1H), 3.77 (d, J = 12.4 Hz, 1H), 3.70 - 3.53 (m, 3H), 3.08 (td, J = 12.1, 3.6 Hz, 1H), 2.76 (dd, J = 12.5, 10.4 Hz, 1H), 1.15 (d, J = 6.2 Hz, 3H).

[0491]

[0492] Step 12-2: Preparation of N-(2-(2-methylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (Compound 12)

[0493]

[0494]

[0495] In the same manner as in Step 1-2, 2-(2-methylmorpholino)benzo[d]thiazol-6-amine (50 mg, 0.20 mmol), sodium nitrite (21 mg, 0.30 mmol) and 1.0 M aqueous hydrochloric acid solution (0.6 mL, 0.60 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (26 mg, 0.40 mmol) and aqueous sodium hydroxide solution to obtain 40 mg (yield 60%) of the title compound.

[0496] 1 H NMR (400 MHz, DMSO-d) δ13.10 (s, 1H), 7.92 (d, J = 2.1 Hz, 1H), 7.52 - 7.39 (m, 2H), 3.99 - 3.87 (m, 2H), 3.80 (d, J = 12.7 Hz, 1H), 3.69 - 3.57 (m, 2H), 3.22 (td, J = 12.2, 3.6 Hz, 1H), 2.90 (dd, J = 12.7, 10.5 Hz, 1H), 1.17 (d, J = 6.2 Hz, 3H).

[0497]

[0498]

[0499] Example 13: Preparation of N-(2-(3-methylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (Compound 13)

[0500] Step 13-1: Preparation of 2-(3-methylmorpholino)benzo[d]thiazol-6-amine (Compound 13-1)

[0501]

[0502] In the same manner as in step 9-1, 2-chloro-6-benzothiazolamine (100 mg, 0.54 mmol) was dissolved in toluene, 3-methylmorpholine (0.57 mL, 5.42 mmol) was added, and the mixture was reacted at 180°C for 12 hours using microwave irradiation, to obtain 108 mg (yield 74%) of the title compound.

[0503] 1 H NMR (400 MHz, DMSO-d6) δ7.17 (d, J = 8.5 Hz, 1H), 6.90 (d, J = 2.3 Hz, 1H), 6.68 - 6.41 (m, 1H), 4.91 (s, 3H), 3.99 - 3.82 (m, 3H), 3.69 - 3.64 (m, 2H), 3.59 - 3.46 (m, 2H), 3.37 - 3.35 (m, 1H), 1.24 (d, J = 6.7 Hz, 3H).

[0504]

[0505]

[0506] Step 13-2: Preparation of N-(2-(3-methylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (compound 13)

[0507]

[0508]

[0509] In the same manner as in Step 1-2, 2-(3-methylmorpholino)benzo[d]thiazol-6-amine (50 mg, 0.20 mmol), sodium nitrite (21 mg, 0.30 mmol) and 1.0 M hydrochloric acid aqueous solution (0.6 mL, 0.60 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (26 mg, 0.40 mmol) and sodium hydroxide aqueous solution to obtain 14 mg (yield 21%) of the title compound.

[0510] 1H NMR (400 MHz, DMSO-d) δ13.10 (s, 1H), 7.91 (d, J = 2.1 Hz, 1H), 7.54 - 7.37 (m, 2H), 4.08 (d, J = 7.4 Hz, 1H), 4.00 - 3.86 (m, 1H), 3.76 - 3.62 (m, 3H), 3.59 - 3.50 (m, 1H), 3.44 (dd, J = 12.4, 3.5 Hz, 1H), 1.29 (d, J = 6.7 Hz, 3H).

[0511]

[0512]

[0513] Example 14: Preparation of N-(2-(4-methylpiperazin-1yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 14)

[0514]

[0515] Step 14-1: Preparation of 2-(4-methylpiperazin-1yl)-6-nitrobenzo[d]oxazole (Compound 14-1)

[0516]

[0517]

[0518] In the same manner as in step 3-2, 89 mg (yield 67%) of the title compound was obtained through 6-nitrobenzo[d]oxazole-2(3H)-thione (100 mg, 0.51 mmol), 1-methylpiperazine (85 μL, 0.76 mmol), triethylamine (213 μL, 1.53 mmol) and toluene.

[0519] 1 H NMR (400 MHz, DMSO-d6) δ8.29 (d, J = 2.3 Hz, 1H), 8.15 (dd, J = 8.7, 2.3 Hz, 1H), 7.40 (d, J = 8.7 Hz, 1H), 3.69 (t, J = 5.1 Hz, 4H), 2.45 (t, J = 5.2 Hz, 4H), 2.24 (s, 3H).

[0520]

[0521]

[0522] Step 14-2: Preparation of 2-(4-methylpiperazin-1-yl)benzo[d]oxazol-6-amine (Compound 14-2)

[0523]

[0524]

[0525] In the same manner as in step 5-3, 2-(4-methylpiperazin-1yl)-6-nitrobenzo[d]oxazole (100 mg, 0.38 mmol) and ammonium chloride (163 mg, 3.05 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (170 mg, 3.05 mmol) was added to the reaction mixture to obtain 45 mg (yield 51%) of the title compound.

[0526] 1 H NMR (400 MHz, DMSO-d6) δ6.95 (d, J = 8.3 Hz, 1H), 6.61 (d, J = 2.1 Hz, 1H), 6.41 (dd, J = 8.3, 2.1 Hz, 1H), 4.87 (s, 2H), 3.48 (t, J = 5.1) Hz, 4H), 2.39 (t, J = 5.1 Hz, 4H), 2.21 (s, 3H).

[0527]

[0528]

[0529] Step 14-3: Preparation of N-(2-(4-methylpiperazin-1yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 14)

[0530]

[0531]

[0532] In the same manner as in Step 1-2, 2-(4-methylpiperazin-1-yl)benzo[d]oxazol-6-amine (30 mg, 0.13 mmol), sodium nitrite (13 mg, 0.19 mmol) and 1.0 M hydrochloric acid aqueous solution (0.4 mL, 0.39 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (17 mg, 0.26 mmol) and sodium hydroxide aqueous solution to obtain 36 mg (yield 40%) of the title compound.

[0533] 1 H NMR (400 MHz, DMSO-d) δ10.05 (s, 1H), 7.31 (d, J = 1.7 Hz, 1H), 7.26 - 7.17 (m, 2H), 3.79 (s, 4H), 3.22 (s, 4H), 2.76 (s, 3H).

[0534]

[0535]

[0536] Example 15: Preparation of N-(2-(2-methylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 15)

[0537]

[0538] Step 15-1: Preparation of 2-(2-methylmorpholino)-6-nitrobenzo[d]oxazole (Compound 15-1)

[0539]

[0540]

[0541] In the same manner as in step 3-2, 6-nitrobenzo[d]oxazole-2(3H)-thione (300 mg, 1.53 mmol), 2-methylmorpholine (247 μL, 2.29 mmol), triethylamine (639 μL, 4.59 mmol) and toluene were added and reacted at 180°C for 30 minutes using microwave irradiation to obtain 193 mg (yield 72%) of the title compound.

[0542] 1H NMR (400 MHz, Chloroform-d) δ8.19 (dd, J = 8.7, 2.2 Hz, 1H), 8.14 (d, J = 2.2 Hz, 1H), 7.34 (d, J = 8.7 Hz, 1H), 4.23 - 4.01 (m, 3H), 3.83 - 3.64 (m, 2H), 3.35 (ddd, J = 13.1, 11.9, 3.7 Hz, 1H), 3.00 (dd, J = 13.0, 10.6 Hz, 1H), 1.28 (d, J = 6.2 Hz, 3H).

[0543]

[0544] Step 15-2: Preparation of 2-(2-methylmorpholino)benzo[d]oxazol-6-amine (Compound 15-2)

[0545]

[0546]

[0547] In the same manner as in step 5-3, 2-(2-methylmorpholino)-6-nitrobenzo[d]oxazole (200 mg, 0.95 mmol) and ammonium chloride (406 mg, 7.60 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (424 mg, 7.60 mmol) was added to the reaction mixture to obtain 88 mg (yield 40%) of the title compound.

[0548] 1 H NMR (400 MHz, DMSO-d6) δ6.97 (d, J = 8.3 Hz, 1H), 6.62 (d, J = 2.1 Hz, 1H), 6.42 (dd, J = 8.3, 2.1 Hz, 1H), 4.89 (s, 2H), 3.87 (ddd, J = 10.9, 6.3, 2.5 Hz, 2H), 3.82 - 3.73 (m, 1H), 3.62 - 3.53 (m, 2H), 3.07 (td, J = 12.4, 3.6 Hz, 1H), 2.76 (dd, J = 12.7, 10.4 Hz, 1H), 1.14 (d, J = 6.2 Hz, 3H).

[0549]

[0550] Step 15-3: Preparation of N-(2-(2-methylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 15)

[0551]

[0552]

[0553] In the same manner as in Step 1-2, 2-(2-methylmorpholino)benzo[d]oxazol-6-amine (30 mg, 0.13 mmol), sodium nitrite (13 mg, 0.19 mmol) and 1.0 M hydrochloric acid aqueous solution (0.4 mL, 0.39 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (17 mg, 0.26 mmol) and sodium hydroxide aqueous solution to obtain 12 mg (yield 30%) of the title compound.

[0554] 1 H NMR (400 MHz, DMSO-d) δ13.08 (s, 1H), 7.50 (d, J = 1.9 Hz, 1H), 7.38 - 7.28 (m, 2H), 4.15 - 3.76 (m, 3H), 3.71 - 3.53 (m, 2H), 3.20 (td, J = 12.5, 3.7 Hz, 1H), 2.89 (dd, J = 12.8, 10.5 Hz, 1H), 1.15 (d, J = 6.2 Hz, 3H).

[0555] Example 16: Preparation of N-(2-(3-methylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 16)

[0556]

[0557] Step 16-1: Preparation of 2-(3-methylmorpholino)-6-nitrobenzo[d]oxazole (Compound 16-1)

[0558]

[0559]

[0560] In the same manner as in step 3-2, 193 mg (yield 72%) of the title compound was obtained through 6-nitrobenzo[d]oxazole-2(3H)-thione (200 mg, 1.02 mmol), 3-methylmorpholine (161 μL, 1.53 mmol), triethylamine (426 μL, 3.06 mmol) and toluene.

[0561] 1 H NMR (400 MHz, Chloroform-d) δ8.24 - 8.09 (m, 2H), 7.34 (d, J = 8.7 Hz, 1H), 4.43 - 4.29 (m, 1H), 4.09 - 3.87 (m, 2H), 3.80 (s, 1H), 3.73 - 3.52 (m, 2H), 1.47 (d, J = 6.9 Hz, 3H).

[0562]

[0563] Step 16-2: Preparation of 2-(3-methylmorpholino)benzo[d]oxazol-6-amine (Compound 16-2)

[0564]

[0565]

[0566] In the same manner as in Step 5-3, 2-(3-methylmorpholino)-6-nitrobenzo[d]oxazole (100 mg, 0.43 mmol) and ammonium chloride (293 mg, 5.47 mmol) were dissolved in distilled water and tetrahydrofuran, and iron (305 mg, 5.47 mmol) was added to the reaction mixture. After removal of the solvent, the product was used in the next step without purification.

[0567]

[0568] Step 16-3: Preparation of N-(2-(3-methylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 16)

[0569]

[0570]

[0571] In the same manner as in Step 1-2, 2-(3-methylmorpholino)benzo[d]oxazol-6-amine (100 mg, 0.43 mmol), sodium nitrite (44 mg, 0.64 mmol) and 1.0 M aqueous hydrochloric acid solution (1.3 mL, 1.29 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (57 mg, 0.86 mmol) and aqueous sodium hydroxide solution to obtain 29 mg (yield 22%) of the title compound.

[0572] 1 H NMR (400 MHz, Chloroform-d) δ9.72 (s, 1H), 7.41 - 7.30 (m, 2H), 7.05 (dd, J = 8.4, 2.2 Hz, 1H), 4.29 (q, J = 7.1 Hz, 1H), 4.01 (dd, J = 11.0, 3.3 Hz, 1H), 3.92 - 3.85 (m, 1H), 3.79 (d, J = 2.2 Hz, 2H), 3.70 - 3.50 (m, 2H), 1.44 (d, J = 6.8 Hz, 3H).

[0573]

[0574] Example 17: Preparation of N-(2-(4-methoxypiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 17)

[0575]

[0576] Step 17-1: Preparation of 2-(4-methoxypiperidin-1-yl)-6-nitrobenzo[d]oxazole (Compound 17-1)

[0577]

[0578]

[0579] In the same manner as in step 3-2, 170 mg (yield 60%) of the title compound was obtained through 6-nitrobenzo[d]oxazole-2(3H)-thione (200 mg, 1.02 mmol), 3-methoxypiperidine (189 μL, 1.53 mmol), triethylamine (426 μL, 3.06 mmol) and toluene.

[0580] 1 H NMR (400 MHz, DMSO-d6) δ8.28 (d, J = 2.2 Hz, 1H), 8.14 (dd, J = 8.7, 2.3 Hz, 1H), 7.38 (d, J = 8.7 Hz, 1H), 3.89 (ddd, J = 11.4, 6.8, 4.0 Hz, 2H), 3.52 (dddd, J = 19.1, 11.5, 8.1, 3.7 Hz, 3H), 3.30 (s, 3H), 2.04 - 1.89 (m, 2H), 1.60 (dtd, J = 12.5, 8.1, 4.0 Hz, 2H).

[0581]

[0582]

[0583] Step 17-2: Preparation of 2-(4-methoxypiperidin-1-yl)benzo[d]oxazol-6-amine (Compound 17-2)

[0584]

[0585]

[0586] In the same manner as in step 5-3, 2-(4-methoxypiperidin-1-yl)-6-nitrobenzo[d]oxazole (120 mg, 0.51 mmol) and ammonium chloride (220 mg, 4.11 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (230 mg, 4.11 mmol) was added to the reaction mixture to obtain 63 mg (yield 49%) of the title compound.

[0587] 1H NMR (400 MHz, Chloroform-d) δ7.13 (d, J = 8.3 Hz, 1H), 6.66 (d, J = 2.2 Hz, 1H), 6.53 (dd, J = 8.2, 2.2 Hz, 1H), 3.91 (ddd, J = 13.1, 6.9, 3.9 Hz, 2H), 3.62 (s, 2H), 3.50 - 3.37 (m, 8H), 1.96 (ddt, J = 13.9, 7.7, 3.7 Hz, 2H), 1.70 (dtd, J = 12.6, 8.1, 3.9 Hz, 2H).

[0588]

[0589]

[0590] Step 17-3: Preparation of N-(2-(4-methoxypiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 17)

[0591]

[0592]

[0593] In the same manner as in Step 1-2, 2-(4-methoxypiperidin-1-yl)benzo[d]oxazol-6-amine (50 mg, 0.20 mmol), sodium nitrite (21 mg, 0.30 mmol) and 1.0 M aqueous hydrochloric acid solution (0.6 mL, 0.61 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (27 mg, 0.40 mmol) and aqueous sodium hydroxide solution to obtain 18 mg (yield 28%) of the title compound.

[0594] 1 H NMR (400 MHz, DMSO-d6) δ13.07 (s, 1H), 7.49 (s, 1H), 7.34 - 7.25 (m, 2H), 3.93 - 3.75 (m, 2H), 3.55 - 3.34 (m, 3H), 3.29 (s, 3H), 2.02 - 1.87 (m, 2H), 1.58 - 1.53 (m, 2H).

[0595]

[0596]

[0597] Example 18: Preparation of N-(2-(2,6-dimethylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0598]

[0599] Step 18-1: Preparation of 2-(2,6-dimethylmorpholino)-6-nitrobenzo[d]oxazole (Compound 18-1)

[0600]

[0601]

[0602] In the same manner as in step 3-2, 227 mg (yield 80%) of the title compound was obtained through 6-nitrobenzo[d]oxazole-2(3H)-thione (200 mg, 1.02 mmol), 2,6-dimethylmorpholine (188 μL, 1.53 mmol), triethylamine (426 μL, 3.06 mmol) and toluene.

[0603] 1 H NMR (400 MHz, Chloroform-d) δ8.19 (dd, J = 8.7, 2.2 Hz, 1H), 8.14 (d, J = 2.1 Hz, 1H), 7.33 (d, J = 8.7 Hz, 1H), 4.15 (dd, J = 13.5, 2.2 Hz, 2H), 3.74 (dtd, J = 12.5, 6.3, 2.6 Hz, 2H), 2.91 (dd, J = 13.1, 10.7 Hz, 2H), 1.29 (d, J = 6.2 Hz, 6H).

[0604]

[0605]

[0606] Step 18-2: Preparation of 2-(2,6-dimethylmorpholino)benzo[d]oxazol-6-amine (Compound 18-2)

[0607]

[0608]

[0609] In the same manner as in step 5-3, 2-(2,6-dimethylmorpholino)-6-nitrobenzo[d]oxazole (200 mg, 0.72 mmol) and ammonium chloride (309 mg, 5.77 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (322 mg, 5.77 mmol) was added to the reaction mixture to obtain 54 mg (yield 30%) of the title compound.

[0610] 1 H NMR (400 MHz, Chloroform-d) δ7.14 (d, J = 8.3 Hz, 1H), 6.67 (d, J = 2.2 Hz, 1H), 6.55 (dd, J = 8.3, 2.2 Hz, 1H), 4.00 (dd, J = 13.3, 2.3 Hz, 2H), 3.73 (dtd, J = 12.4, 6.2, 2.5 Hz, 2H), 3.60 (s, 2H), 2.78 (dd, J = 12.9, 10.6 Hz, 2H), 1.25 (d, J = 6.2 Hz, 6H).

[0611]

[0612]

[0613]

[0614] Step 18-3: Preparation of N-(2-(2,6-dimethylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 18)

[0615]

[0616]

[0617] In the same manner as in Step 1-2, 2-(2,6-dimethylmorpholino)benzo[d]oxazol-6-amine (50 mg, 0.20 mmol), sodium nitrite (21 mg, 0.30 mmol) and 1.0 M aqueous hydrochloric acid solution (0.6 mL, 0.61 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (27 mg, 0.40 mmol) and aqueous sodium hydroxide solution to obtain 19 mg (yield 29%) of the title compound.

[0618] 1H NMR (400 MHz, Chloroform-d) δ9.76 (s, 1H), 7.38 - 7.30 (m, 2H), 7.06 (dd, J = 8.4, 2.2 Hz, 1H), 4.18 - 4.00 (m, 3H), 3.74 (ddp, J = 12.4, 6.1, 3.1, 2.5 Hz, 2H), 2.86 (dd, J = 13.0, 10.7 Hz, 2H), 1.27 (d, J = 6.2 Hz, 8H).

[0619]

[0620] Example 19: Preparation of N-(5-chloro-2-morpholinobenzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0621]

[0622] Step 19-1: Preparation of 5-chloro-6-nitrobenzo[d]oxazole-2(3H)-thione (Compound 19-1)

[0623]

[0624]

[0625] 2-Amino-4-chloro-5-nitrophenol (200 mg, 1.06 mmol) was dissolved in pyridine, potassium ethyl xanthogate (187 mg, 1.17 mmol) was added, and the mixture was stirred at 120°C for 2 hours. After the reaction was completed, the mixture was extracted with distilled water and ethyl acetate, and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and used in the next step without purification.

[0626]

[0627] Step 19-2: Preparation of 5-chloro-2-morpholino-6-nitrobenzo[d]oxazole (Compound 19-2)

[0628]

[0629]

[0630] In the same manner as in step 3-2, 30 mg (yield 24%) of the title compound was obtained from 5-chloro-6-nitrobenzo[d]oxazole-2(3H)-thione (100 mg, 0.43 mmol), morpholine (56 μL, 0.65 mmol), triethylamine (181 μL, 1.30 mmol) and toluene.

[0631] 1 H NMR (400 MHz, DMSO-d6) δ8.29 (s, 1H), 7.61 (s, 1H), 3.76 - 3.74 (m, 4H), 3.69 - 3.67 (m, 4H).

[0632]

[0633] Step 19-3: Preparation of 5-chloro-2-morpholinobenzo[d]oxazol-6-amine (Compound 19-3)

[0634]

[0635]

[0636] In the same manner as in Step 3-3, 5-chloro-2-morpholino-6-nitrobenzo[d]oxazole (50 mg, 0.18 mmol) and 10% Pd / C (37 mg, 0.03 mmol) were dissolved in 1,4-dioxane and purified by hydrogen to give the title compound, which was used in the next step without purification.

[0637]

[0638] Step 19-4: Preparation of N-(5-chloro-2-morpholinobenzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 19)

[0639]

[0640]

[0641] In the same manner as in Step 1-2, 5-chloro-2-morpholinobenzo[d]oxazol-6-amine (30 mg, 0.12 mmol), sodium nitrite (12 mg, 0.18 mmol) and 1.0 M hydrochloric acid aqueous solution (0.3 mL, 0.35 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (16 mg, 0.24 mmol) and sodium hydroxide aqueous solution to obtain 11 mg (yield 27%) of the title compound.

[0642] 1 H NMR (400 MHz, Chloroform-d) δ10.10 (s, 1H), 7.55 (s, 1H), 7.35 (s, 1H), 3.83 (t,J= 4.8 Hz, 4H), 3.71 (t,J= 4.9 Hz, 4H).

[0643]

[0644]

[0645]

[0646] Example 20: Preparation of N-(2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0647]

[0648] Step 20-1: Preparation of 2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-6-nitrobenzo[d]oxazole (Compound 20-1)

[0649]

[0650]

[0651] In the same manner as in step 9-1, 799 mg (yield 57%) of the title compound was obtained through 6-nitrobenzo[d]oxazole-2(3H)-thione (1 g, 5.10 mmol), 3-oxa-8-azabicyclo[3.2.1]octane hydrochloride (1.14 g, 7.65 mmol), triethylamine (2.13 mL, 15.29 mmol) and toluene.

[0652] 1H NMR (400 MHz, Chloroform-d) δ8.20 (dd, J = 8.7, 2.2 Hz, 1H), 8.15 (d, J = 2.2 Hz, 1H), 7.35 (d, J = 8.7 Hz, 1H), 4.49 (dd, J = 4.6, 2.2 Hz, 2H), 3.87 (d, J = 11.2 Hz, 2H), 3.72 (d, J = 11.6 Hz, 2H), 2.25 - 2.09 (m, 4H).

[0653]

[0654] Step 20-2: Preparation of 2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]oxazol-6-amine (Compound 20-2)

[0655]

[0656]

[0657] In the same manner as in step 5-3, 2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-6-nitrobenzo[d]oxazole (750 mg, 2.72 mmol) and ammonium chloride (1.17 g, 21.80 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (1.22 g, 21.80 mmol) was added to the reaction mixture to obtain 241 mg (yield 36%) of the title compound.

[0658] 1 H NMR (400 MHz, Chloroform-d) δ7.17 (d, J = 8.3 Hz, 1H), 6.68 (d, J = 2.2 Hz, 1H), 6.56 (dd, J = 8.2, 2.2 Hz, 1H), 4.34 (s, 2H), 3.89 (d, J = 11.0) Hz, 2H), 3.63 (d, J = 10.8 Hz, 5H), 2.13 - 2.02 (m, 4H).

[0659]

[0660] Step 20-3: Preparation of N-(2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 20)

[0661]

[0662]

[0663] In the same manner as in Step 1-2, 2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]oxazol-6-amine (230 mg, 0.94 mmol), sodium nitrite (97 mg, 1.41 mmol) and 1.0 M aqueous hydrochloric acid solution (2.8 mL, 2.81 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (124 mg, 1.88 mmol) and aqueous sodium hydroxide solution to obtain 235 mg (yield 78%) of the title compound.

[0664] 1 H NMR (400 MHz, DMSO-d6) δ13.08 (s, 2H), 7.52 (s, 1H), 7.34 (s, 2H), 4.35 (s, 2H), 3.72 (d, J = 11.0 Hz, 2H), 3.61 (d, J = 11.1 Hz, 2H), 2.00 (d, J = 2.2 Hz, 4H).

[0665]

[0666]

[0667]

[0668] Example 21: Preparation of tert-butyl 4-(6-(2-(dicyano-methylene)hydrazinyl)benzo[d]oxazol-2-yl)piperazine-1-carboxylate

[0669]

[0670] Step 21-1: Preparation of tert-butyl 4-(6-nitrobenzo[d]oxazol-2-yl)piperazine-1-carboxylate (Compound 21-1)

[0671]

[0672]

[0673] In the same manner as in step 9-1, 246 mg (yield 53%) of the title compound was obtained through 6-nitrobenzo[d]oxazole-2(3H)-thione (300 mg, 1.53 mmol), 1-Boc-piperazine (427 mg, 2.29 mmol), triethylamine (639 μL, 4.59 mmol) and toluene.

[0674] 1 H NMR (400 MHz, Chloroform-d) δ8.19 (dd, J = 8.7, 2.2 Hz, 1H), 8.15 (d, J = 2.2 Hz, 1H), 7.34 (d, J = 8.7 Hz, 1H), 3.76 (dd, J = 6.6, 4.0 Hz, 4H), 3.61 (dd, J = 6.4, 4.0 Hz, 4H), 1.50 (s, 9H).

[0675]

[0676]

[0677] Step 21-2: Preparation of tert-butyl 4-(6-aminobenzo[d]oxazol-2-yl)piperazine-1-carboxylate (Compound 21-2)

[0678]

[0679]

[0680] In the same manner as in step 3-3, tert-butyl 4-(6-nitrobenzo[d]oxazol-2-yl)piperazine-1-carboxylate (100 mg, 0.29 mmol) and 10% Pd / C (61 mg, 0.06 mmol) were dissolved in 1,4-dioxane, and 43 mg (yield 47%) of the title compound was obtained through hydrogen.

[0681] 1 H NMR (400 MHz, Chloroform-d) δ7.15 (d, J = 8.3 Hz, 1H), 6.68 (d, J = 2.2 Hz, 1H), 6.55 (dd, J = 8.3, 2.2 Hz, 1H), 3.64 - 3.52 (m, 10H), 1.49 (s, 9H).

[0682]

[0683]

[0684] Step 21-3: Preparation of tert-butyl 4-(6-(2-(dicyano-methylene)hydrazinyl)benzo[d]oxazol-2-yl)piperazine-1-carboxylate (compound 21)

[0685]

[0686]

[0687] In the same manner as in Step 1-2, tert-butyl 4-(6-aminobenzo[d]oxazol-2-yl)piperazine-1-carboxylate (30 mg, 0.09 mmol), sodium nitrite (10 mg, 0.14 mmol) and 1.0 M aqueous hydrochloric acid solution (0.3 mL, 0.28 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (12 mg, 0.19 mmol) and aqueous sodium hydroxide solution to obtain 36 mg (yield 96%) of the title compound.

[0688] 1 H NMR (400 MHz, Chloroform-d) δ9.79 (s, 1H), 7.37 (d, J = 2.1 Hz, 1H), 7.33 (d, J = 8.4 Hz, 1H), 7.07 (dd, J = 8.4, 2.2 Hz, 1H), 3.69 (dd, J = 6.7, 3.8 Hz, 4H), 3.59 (dd, J = 6.5, 3.8 Hz, 4H), 1.50 (s, 9H).

[0689]

[0690] Example 22: Preparation of tert-butyl 4-(6-(2-(dicyano-methylene)hydrazinyl)benzo[d]thiazol-2-yl)piperazine-1-carboxylate

[0691] Step 22-1: Preparation of tert-butyl 4-(6-aminobenzo[d]thiazol-2-yl)piperazine-1-carboxylate (Compound 22-1)

[0692]

[0693]

[0694] In the same manner as in step 9-1, 223 mg (yield 62%) of the title compound was obtained through 2-chloro-6-benzothiazolamine (200 mg, 1.08 mmol), 1-Boc-piperazine (403 mg, 2.17 mmol), triethylamine (453 μL, 3.20 mmol) and toluene.

[0695] 1 H NMR (400 MHz, Chloroform-d) δ7.37 (d, J = 8.5 Hz, 1H), 6.95 (d, J = 2.4 Hz, 1H), 6.69 (dd, J = 8.5, 2.4 Hz, 1H), 3.62 - 3.48 (m, 10H), 1.48 (s, 9H).

[0696]

[0697]

[0698] Step 22-2: Preparation of tert-butyl 4-(6-(2-(dicyano-methylene)hydrazinyl)benzo[d]thiazol-2-yl)piperazine-1-carboxylate (compound 22)

[0699]

[0700]

[0701] In the same manner as in Step 1-2, tert-butyl 4-(6-aminobenzo[d]thiazol-2-yl)piperazine-1-carboxylate (200 mg, 0.60 mmol), sodium nitrite (62 mg, 0.90 mmol) and 1.0 M aqueous hydrochloric acid solution (1.8 mL, 1.79 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (79 mg, 1.20 mmol) and aqueous sodium hydroxide solution to obtain 24 mg (yield 98%) of the title compound.

[0702] 1H NMR (400 MHz, Chloroform-d) δ9.95 (s, 1H), 7.66 (d, J = 2.3 Hz, 1H), 7.54 (d, J = 8.7 Hz, 1H), 7.22 (dd, J = 8.7, 2.3 Hz, 1H), 3.68 - 3.50 (m, 8H), 1.49 (s, 8H).

[0703]

[0704] Example 23: Preparation of N-(2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0705] Step 23-1: Preparation of 2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]thiazol-6-amine (Compound 23-1)

[0706]

[0707] In the same manner as in step 9-1, 110 mg (yield 39%) of the title compound was obtained through 2-chloro-6-benzothiazolamine (200 mg, 1.08 mmol), 8-oxa-3-azabicyclo[3.2.1]octane hydrochloride (324 mg, 2.17 mmol), triethylamine (453 μL, 3.25 mmol) and toluene.

[0708] 1 H NMR (400 MHz, Chloroform-d) δ7.36 (d, J = 8.5 Hz, 1H), 6.95 (d, J = 2.4 Hz, 1H), 6.69 (dd, J = 8.5, 2.4 Hz, 1H), 4.47 (d, J = 2.7 Hz, 2H), 3.64 - 3.54 (m, 4H), 3.41 (dd, J = 12.2, 2.6 Hz, 2H), 2.07 - 1.81 (m, 4H).

[0709]

[0710]

[0711] Step 23-2: Preparation of N-(2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (compound 23)

[0712]

[0713]

[0714] In the same manner as in Step 1-2, 2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]thiazol-6-amine (100 mg, 0.38 mmol), sodium nitrite (40 mg, 0.57 mmol) and 1.0 M aqueous hydrochloric acid solution (1.1 mL, 1.15 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (51 mg, 0.76 mmol) and aqueous sodium hydroxide solution to obtain 117 mg (yield 91%) of the title compound.

[0715] 1 H NMR (400 MHz, DMSO-d6) δ13.11 (s, 1H), 7.90 (d, J = 2.0 Hz, 1H), 7.49 - 7.38 (m, 2H), 4.45 (d, J = 4.8 Hz, 2H), 3.63 (d, J = 12.2 Hz, 2H), 2.00 - 1.68 (m, 4H).

[0716]

[0717] Example 24: Preparation of N-(2-(piperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0718] Step 24-1: Preparation of 2-(piperazin-1-yl)benzo[d]thiazol-6-amine (compound 24-1)

[0719]

[0720] In the same manner as in step 9-1, 90 mg (yield 35%) of the title compound was obtained through 2-chloro-6-benzothiazolamine (200 mg, 1.08 mmol), piperazine (187 mg, 2.17 mmol) and toluene.

[0721] 1 H NMR (400 MHz, Chloroform-d) δ7.36 (d, J = 8.5 Hz, 1H), 6.95 (d, J = 2.4 Hz, 1H), 6.68 (dd, J = 8.5, 2.4 Hz, 1H), 3.63 - 3.46 (m, 6H), 3.03 - 2.93 (m, 4H).

[0722]

[0723]

[0724] Step 24-2: Preparation of N-(2-(piperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (compound 24)

[0725]

[0726]

[0727] In the same manner as in Step 1-2, 2-(piperazin-1-yl)benzo[d]thiazol-6-amine (100 mg, 0.43 mmol), sodium nitrite (44 mg, 0.64 mmol) and 1.0 M hydrochloric acid aqueous solution (1.3 mL, 1.28 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (56 mg, 0.85 mmol) and sodium hydroxide aqueous solution to obtain 42 mg (yield 31%) of the title compound.

[0728] 1 H NMR (400 MHz, Chloroform-d) δ9.76 (s, 1H), 7.70 (d, J = 2.3 Hz, 1H), 7.59 (d, J = 8.7 Hz, 1H), 7.26 - 7.21 (m, 1H), 4.51 - 4.44 (m, 2H), 4.00 (dd, J = 6.5, 4.6 Hz, 2H), 3.94 (dd, J = 6.3, 4.5 Hz, 2H), 3.70 (dd, J = 6.5, 4.6 Hz, 2H).

[0729]

[0730] Example 25: Preparation of N-(2-(piperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0731]

[0732] Step 25-1: Preparation of 6-nitro-2-(piperazin-1-yl)benzo[d]oxazole (Compound 25-1)

[0733]

[0734]

[0735] In the same manner as in step 9-1, 124 mg (yield 33%) of the title compound was obtained from 6-nitrobenzo[d]oxazole-2(3H)-thione (300 mg, 1.53 mmol), piperazine (198 mg, 2.29 mmol), triethylamine (639 μL, 4.59 mmol) and toluene.

[0736] 1 H NMR (400 MHz, DMSO-d6) δ8.27 (d, J = 2.3 Hz, 1H), 8.14 (dd, J = 8.7, 2.3 Hz, 1H), 7.38 (d, J = 8.7 Hz, 1H), 3.61 (t, J = 5.1 Hz, 4H), 2.81 (t, J = 5.1 Hz, 4H), 2.65 (s, 2H).

[0737]

[0738]

[0739] Step 25-2: Preparation of 2-(piperazin-1-yl)benzo[d]oxazol-6-amine (compound 25-2)

[0740]

[0741]

[0742] In the same manner as in step 3-3, 6-nitro-2-(piperazin-1-yl)benzo[d]oxazole (100 mg, 0.40 mmol) and 10% Pd / C (86 mg, 0.08 mmol) were dissolved in 1,4-dioxane, and 50 mg (yield 57%) of the title compound was obtained through hydrogen.

[0743] 1H NMR (400 MHz, Chloroform-d) δ7.14 (d, J = 8.3 Hz, 1H), 6.67 (d, J = 2.2 Hz, 1H), 6.54 (dd, J = 8.3, 2.2 Hz, 1H), 3.71 - 3.54 (m, 6H), 3.05 - 2.89 (m, 4H).

[0744]

[0745]

[0746] Step 25-3: Preparation of N-(2-(piperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 25)

[0747]

[0748]

[0749] In the same manner as in Step 1-2, 2-(piperazin-1-yl)benzo[d]oxazol-6-amine (50 mg, 0.23 mmol), sodium nitrite (24 mg, 0.34 mmol) and 1.0 M hydrochloric acid aqueous solution (0.7 mL, 0.69 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (30 mg, 0.46 mmol) and sodium hydroxide aqueous solution to obtain 14 mg (yield 20%) of the title compound.

[0750] 1 H NMR (400 MHz, DMSO-d6) δ13.10 (s, 1H), 7.54 (s, 1H), 7.35 (s, 1H), 4.45 - 4.39 (m, 2H), 3.90 (q, J = 5.1 Hz, 4H), 3.68 (dd, J = 6.6, 4.5 Hz, 2H).

[0751]

[0752] Example 26: Preparation of N-(2-(piperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0753]

[0754] Step 25-1: Preparation of 2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)-6-nitrobenzo[d]oxazole (Compound 25-1)

[0755]

[0756]

[0757] In the same manner as in step 9-1, 142 mg (yield 34%) of the title compound was obtained through 6-nitrobenzo[d]oxazole-2(3H)-thione (300 mg, 1.53 mmol), 8-oxa-3-azabicyclo[3.2.1]octane hydrochloride (343 mg, 2.29 mmol), triethylamine (639 μL, 4.59 mmol) and toluene.

[0758] 1 H NMR (400 MHz, Chloroform-d) δ8.19 (dd, J = 8.7, 2.2 Hz, 1H), 8.14 (d, J = 2.2 Hz, 1H), 7.33 (d, J = 8.7 Hz, 1H), 4.52 (d, J = 2.6 Hz, 2H), 3.91 (d, J = 13.1 Hz, 2H), 3.53 (dd, J = 12.8, 2.5 Hz, 2H), 2.12 - 2.01 (m, 2H), 1.96 - 1.83 (m, 2H).

[0759]

[0760]

[0761] Step 26-2: Preparation of 2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]oxazol-6-amine (Compound 26-2)

[0762]

[0763]

[0764] In the same manner as in step 5-3, 2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)-6-nitrobenzo[d]oxazole (120 mg, 0.44 mmol) and ammonium chloride (187 mg, 3.49 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (195 mg, 3.49 mmol) was added to the reaction mixture to obtain 33 mg (yield 30%) of the title compound.

[0765] 1 H NMR (400 MHz, Chloroform-d) δ7.14 (d, J = 8.2 Hz, 1H), 6.67 (d, J = 2.2 Hz, 1H), 6.54 (dd, J = 8.2, 2.2 Hz, 1H), 4.46 (d, J = 2.7 Hz, 1H), 3.85 - 3.67 (m, 2H), 3.66 - 3.47 (m, 2H), 3.40 (dd, J = 12.5, 2.5 Hz, 2H), 2.15 - 1.75 (m, 4H).

[0766]

[0767]

[0768] Step 26-3: Preparation of N-(2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 26)

[0769]

[0770]

[0771] In the same manner as in Step 1-2, 2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]oxazol-6-amine (20 mg, 0.08 mmol), sodium nitrite (8 mg, 0.12 mmol) and 1.0 M aqueous hydrochloric acid solution (0.2 mL, 0.24 mmol) were dissolved in distilled water, and then the resulting mixture was treated with malononitrile (11 mg, 0.16 mmol) and aqueous sodium hydroxide solution to obtain 6 mg (yield 24%) of the title compound.

[0772] 1H NMR (400 MHz, DMSO-d6) δ13.09 (s, 1H), 7.50 (d, J = 2.0 Hz, 1H), 7.36 - 7.26 (m, 2H), 4.44 (s, 3H), 3.71 (d, J = 12.4 Hz, 3H), 1.98 - 1.65 (m, 6H).

[0773]

[0774] Example 27: Preparation of N-(2-(4-isopropylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0775] Step 27-1: Preparation of 2-(4-isopropylpiperazin-1-yl)benzo[d]thiazol-6-amine (Compound 27-1)

[0776]

[0777] In the same manner as in step 9-1, 238 mg (yield 79%) of the title compound was obtained through 2-chloro-6-benzothiazolamine (200 mg, 1.08 mmol), 1-isopropyl piperazine (310 μL, 2.17 mmol), triethylamine (249 μL, 3.25 mmol) and toluene.

[0778] 1 H NMR (400 MHz, Chloroform-d) δ7.36 (d, J = 8.5 Hz, 1H), 6.95 (d, J = 2.3 Hz, 1H), 6.68 (dd, J = 8.5, 2.4 Hz, 1H), 3.58 (t, J = 5.2 Hz, 6H), 2.75 (p, J = 6.5 Hz, 1H), 2.63 (t, J = 5.1 Hz, 4H), 1.07 (d, J = 6.5 Hz, 6H).

[0779]

[0780]

[0781] Step 27-2: Preparation of N-(2-(4-isopropylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (Compound 27)

[0782]

[0783]

[0784] In the same manner as in Step 1-2, 2-(4-isopropylpiperazin-1-yl)benzo[d]thiazol-6-amine (200 mg, 0.72 mmol), sodium nitrite (75 mg, 1.08 mmol) and 1.0 M aqueous hydrochloric acid solution (2.2 mL, 2.17 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (96 mg, 1.45 mmol) and aqueous sodium hydroxide solution to obtain 51 mg (yield 20%) of the title compound.

[0785] 1 H NMR (400 MHz, Chloroform-d) δ7.64 (d, J = 2.3 Hz, 1H), 7.52 (d, J = 8.7 Hz, 1H), 7.19 (dd, J = 8.7, 2.4 Hz, 1H), 3.66 (t, J = 5.1 Hz, 4H), 2.78 (p, J = 6.5 Hz, 1H), 2.65 (t, J = 5.2 Hz, 4H), 1.08 (d, J = 6.5 Hz, 6H).

[0786]

[0787] Example 28: Preparation of N-(2-(piperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0788]

[0789] Step 28-1: Preparation of 2-(4-isopropylpiperazin-1-yl)-6-nitrobenzo[d]oxazole (Compound 28-1)

[0790]

[0791]

[0792] In the same manner as in step 9-1, 267 mg (yield 49%) of the title compound was obtained through 6-nitrobenzo[d]oxazole-2(3H)-thione (300 mg, 1.53 mmol), 1-isopropyl piperazine (328 μL, 2.29 mmol), triethylamine (351 μL, 4.59 mmol) and toluene.

[0793] 1 H NMR (400 MHz, Chloroform-d) δ8.18 (dd, J = 8.7, 2.2 Hz, 1H), 8.13 (d, J = 2.2 Hz, 1H), 7.31 (d, J = 8.7 Hz, 1H), 3.79 (t, 4H), 2.80 (p, J = 6.6) Hz, 1H), 2.66 (t, 4H), 1.08 (d, J = 6.5 Hz, 6H).

[0794]

[0795]

[0796] Step 28-2: Preparation of 2-(4-isopropylpiperazin-1-yl)benzo[d]oxazol-6-amine (Compound 28-2)

[0797]

[0798]

[0799] In the same manner as in step 3-3, 2-(4-isopropylpiperazin-1-yl)-6-nitrobenzo[d]oxazole (200 mg, 0.69 mmol) and 10% Pd / C (147 mg, 0.14 mmol) were dissolved in 1,4-dioxane, and 148 mg (yield 82%) of the title compound was obtained through hydrogen.

[0800] 1H NMR (400 MHz, Chloroform-d) δ7.14 (d, J = 8.3 Hz, 1H), 6.67 (d, J = 2.2 Hz, 1H), 6.53 (dd, J = 8.3, 2.2 Hz, 1H), 3.69 - 3.61 (m, 4H), 3.57 (s, 2H), 2.75 (p, J = 6.6 Hz, 1H), 2.62 (t, J = 5.1 Hz, 4H), 1.07 (d, J = 6.5 Hz, 6H).

[0801]

[0802]

[0803] Step 28-3: Preparation of N-(2-(4-isopropylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 28)

[0804]

[0805]

[0806] In the same manner as in Step 1-2, 2-(4-isopropylpiperazin-1-yl)benzo[d]oxazol-6-amine (100 mg, 0.38 mmol), sodium nitrite (40 mg, 0.58 mmol) and 1.0 M aqueous hydrochloric acid solution (1.1 mL, 1.15 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (51 mg, 0.77 mmol) and aqueous sodium hydroxide solution to obtain 123 mg (yield 95%) of the title compound.

[0807] 1 H NMR (400 MHz, Acetone-d6) δ7.51 (d, J = 2.1 Hz, 1H), 7.35 (dd, J = 8.5, 2.1 Hz, 1H), 7.26 (d, J = 8.5 Hz, 1H), 3.72 - 3.68 (m, 4H), 2.85 (p, J = 6.6 Hz, 1H), 2.74 - 2.67 (m, 4H), 1.08 (d, J = 6.6 Hz, 6H).

[0808]

[0809] Example 29: Preparation of tert-butyl (2S,6R)-4-(6-(2-(dicyano-methylene)hydrazinyl)benzo[d]thiazol-2-yl)-2,6-dimethylpiperazine-1-carboxylate

[0810] Step 29-1: Preparation of tert-butyl (2S,6R)-4-(6-aminobenzo[d]thiazol-2-yl)-2,6-dimethylpiperazine-1-carboxylate (Compound 29-1)

[0811]

[0812] In the same manner as in Step 9-1, 76 mg (yield 13%) of the title compound was obtained through 2-chloro-6-benzothiazolamine (300 mg, 1.62 mmol), tert-butyl (2S,6R)-2,6-dimethylpiperazine-1-carboxylate (696 mg, 3.25 mmol), triethylamine (679 μL, 4.87 mmol) and toluene.

[0813] 1 H NMR (400 MHz, DMSO-d6) δ7.16 (d, J = 8.5 Hz, 1H), 6.89 (d, J = 2.3 Hz, 1H), 6.56 (dd, J = 8.5, 2.3 Hz, 1H), 4.90 (s, 2H), 4.15 (p, J = 6.6) Hz, 2H), 3.78 (d, J = 12.9 Hz, 2H), 3.22 (dd, J = 12.9, 4.5 Hz, 2H), 1.43 (s, 8H), 1.19 (d, J = 6.8 Hz, 6H).

[0814]

[0815]

[0816] Step 29-2: Preparation of tert-butyl (2S,6R)-4-(6-(2-(dicyano-methylene)hydrazinyl)benzo[d]thiazol-2-yl)-2,6-dimethylpiperazine-1-carboxylate (compound 29)

[0817]

[0818]

[0819] In the same manner as in Step 1-2, tert-butyl (2S,6R)-4-(6-aminobenzo[d]thiazol-2-yl)-2,6-dimethylpiperazine-1-carboxylate (60 mg, 0.17 mmol), sodium nitrite (17 mg, 0.25 mmol) and 1.0 M hydrochloric acid aqueous solution (0.5 mL, 0.50 mmol) were dissolved in distilled water, and then the resulting mixture was treated with malononitrile (22 mg, 0.33 mmol) and sodium hydroxide aqueous solution to obtain 61 mg (yield 84%) of the title compound.

[0820] 1 H NMR (400 MHz, Chloroform-d) δ7.64 (d, J = 2.3 Hz, 1H), 7.52 (d, J = 8.7 Hz, 1H), 7.21 (dd, J = 8.7, 2.4 Hz, 1H), 4.41 - 4.25 (m, 2H), 3.94 (d, J = 13.0 Hz, 2H), 3.39 (dd, J = 13.1, 4.7 Hz, 2H), 1.50 (s, 9H), 1.32 (d, J = 6.9 Hz, 6H).

[0821]

[0822] Example 30: Preparation of N-(2-(4-isopropylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0823] Step 30-1: Preparation of 1-(4-(6-aminobenzo[d]thiazol-2-yl)piperazin-1-yl)ethan-1-one (compound 30-1)

[0824]

[0825] In the same manner as in step 9-1, 208 mg (yield 69%) of the title compound was obtained through 2-chloro-6-benzothiazolamine (200 mg, 1.08 mmol), 1-acetyl piperazine (278 mg, 2.17 mmol), triethylamine (453 μL, 3.25 mmol) and toluene.

[0826] 1H NMR (400 MHz, Chloroform-d) δ7.38 (d, J = 8.5 Hz, 1H), 6.97 (d, J = 2.3 Hz, 1H), 6.70 (dd, J = 8.5, 2.3 Hz, 1H), 3.76 (d, J = 5.5 Hz, 2H), 3.66 - 3.49 (m, 8H), 2.15 (s, 3H).

[0827]

[0828] Step 30-2: Preparation of N-(2-(4-acetylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (compound 30)

[0829]

[0830]

[0831] In the same manner as in Step 1-2, 1-(4-(6-aminobenzo[d]thiazol-2-yl)piperazin-1-yl)ethan-1-one (180 mg, 0.65 mmol), sodium nitrite (67 mg, 0.98 mmol) and 1.0 M aqueous hydrochloric acid solution (2.0 mL, 1.95 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (86 mg, 1.30 mmol) and aqueous sodium hydroxide solution to obtain 117 mg (yield 51%) of the title compound.

[0832] 1 H NMR (400 MHz, DMSO-d6) δ13.10 (s, 1H), 7.91 (d, J = 2.2 Hz, 1H), 7.53 - 7.36 (m, 2H), 3.64 - 3.52 (m, 8H), 2.06 (s, 3H).

[0833]

[0834] Example 31: Preparation of N-(2-(4-acetylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide

[0835]

[0836] Step 31-1: Preparation of 1-(4-(6-nitrobenzo[d]oxazol-2-yl)piperazin-1-yl)ethan-1-one (compound 31-1)

[0837]

[0838]

[0839] In the same manner as in step 9-1, 153 mg (yield 34%) of the title compound was obtained through 6-nitrobenzo[d]oxazole-2(3H)-thione (300 mg, 1.53 mmol), 1-acetyl piperazine (392 mg, 3.06 mmol), triethylamine (639 μL, 4.59 mmol) and toluene.

[0840] 1 H NMR (400 MHz, Chloroform-d) δ8.23 - 8.15 (m, 2H), 7.36 (d, J = 8.6 Hz, 1H), 3.89 - 3.73 (m, 6H), 3.69 - 3.57 (m, 2H), 2.18 (s, 3H).

[0841]

[0842] Step 31-2: Preparation of 1-(4-(6-aminobenzo[d]oxazol-2-yl)piperazin-1-yl)ethan-1-one (Compound 31-2)

[0843]

[0844]

[0845] In the same manner as in step 5-3, 1-(4-(6-nitrobenzo[d]oxazol-2-yl)piperazin-1-yl)ethan-1-one (120 mg, 0.41 mmol) and ammonium chloride (177 mg, 3.31 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (185 mg, 3.31 mmol) was added to the reaction mixture to obtain 33 mg (yield 31%) of the title compound.

[0846] 1H NMR (400 MHz, DMSO-d6) δ6.98 (d, J = 8.3 Hz, 1H), 6.63 (d, J = 2.2 Hz, 1H), 6.42 (dd, J = 8.3, 2.2 Hz, 1H), 4.90 (s, 2H), 3.63 - 3.40 (m, 8H), 2.05 (s, 3H).

[0847]

[0848]

[0849] Step 31-3: Preparation of N-(2-(4-acetylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 31)

[0850]

[0851]

[0852] In the same manner as in Step 1-2, 1-(4-(6-aminobenzo[d]oxazol-2-yl)piperazin-1-yl)ethan-1-one (20 mg, 0.08 mmol), sodium nitrite (8 mg, 0.12 mmol) and 1.0 M hydrochloric acid aqueous solution (0.2 mL, 0.23 mmol) were dissolved in distilled water, and then the title compound (25 mg, 98% yield) was obtained through malononitrile (10 mg, 0.15 mmol) and sodium hydroxide aqueous solution.

[0853] 1 H NMR (400 MHz, Chloroform-d) δ10.12 (s, 1H), 7.42 (d, J = 2.1 Hz, 1H), 7.35 (d, J = 8.4 Hz, 1H), 7.13 (dd, J = 8.4, 2.2 Hz, 1H), 3.84 - 3.69 (m, 6H), 3.70 - 3.58 (m, 2H), 2.18 (s, 3H).

[0854]

[0855] Example 32: Preparation of N-(1-ethyl-2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide (Compound 32)

[0856] Step 32-1: 4-(1-Ethyl-6-nitro-1H-benzo[d]imidazol-2-yl)morpholine (Compound 32-1)

[0857] 4-(1-Ethyl-5-nitro-1H-benzo[d]imidazol-2-yl)morpholine (Compound 32-2)

[0858]

[0859]

[0860]

[0861] In the same manner as in Step 5-1, 43 mg (yield 19%) of the title compound (32-1) and 30 mg (yield 14%) of the title compound (32-2) were obtained from 4-(6-nitro-1H-benzo[d]imidazol-2-yl)morpholine (200 mg, 0.81 mmol), DBU (361 μL, 2.42 mmol), ethyl iodide (194 μL, 2.42 mmol) and dimethylformamide.

[0862] Compound 32-1: 1 H NMR (400 MHz, Chloroform-d) δ8.21 - 8.11 (m, 2H), 7.58 (d, J = 8.7 Hz, 1H), 4.13 (q, J = 7.3 Hz, 2H), 3.94 - 3.86 (m, 4H), 3.48 - 3.38 (m, 4H), 1.53 (t, J = 7.3 Hz, 3H).

[0863] Compound 32-2: 1 H NMR (400 MHz, Chloroform-d) δ8.48 (d, J = 2.2 Hz, 1H), 8.13 (dd, J = 8.9, 2.2 Hz, 1H), 7.28 - 7.25 (m, 1H), 4.19 - 4.07 (m, 2H), 3.94 - 3.86 (m, 4H), 3.40 - 3.34 (m, 4H), 1.50 (t, J = 7.3 Hz, 3H).

[0864]

[0865] Step 32-2: Preparation of 1-ethyl-2-morpholino-1H-benzo[d]imidazol-6-amine (compound 32-3)

[0866]

[0867]

[0868] In the same manner as in step 5-3, 4-(1-ethyl-6-nitro-1H-benzo[d]imidazol-2-yl)morpholine (70 mg, 0.25 mmol) and ammonium chloride (108 mg, 2.03 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (113 mg, 2.03 mmol) was added to the reaction mixture to obtain 34 mg (yield 54%) of the title compound.

[0869] 1 H NMR (400 MHz, DMSO-d6) δ7.10 (d, J = 8.4 Hz, 1H), 6.52 (dd, J = 2.1, 0.6 Hz, 1H), 6.42 (dd, J = 8.4, 2.1 Hz, 1H), 4.79 (s, 2H), 3.94 (q, J = 7.2 Hz, 2H), 3.81 - 3.68 (m, 4H), 3.12 - 2.99 (m, 4H), 1.36 (s, 2H), 1.30 (t, J = 7.2 Hz, 3H).

[0870]

[0871]

[0872] Step 32-3: Preparation of N-(1-ethyl-2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide (compound 32)

[0873]

[0874]

[0875] In the same manner as in Step 1-2, 1-ethyl-2-morpholino-1H-benzo[d]imidazol-6-amine (30 mg, 0.12 mmol), sodium nitrite (13 mg, 0.18 mmol) and 1.0 M hydrochloric acid aqueous solution (0.4 mL, 0.36 mmol) were dissolved in distilled water, and then the resulting mixture was treated with malononitrile (16 mg, 0.24 mmol) and sodium hydroxide aqueous solution to obtain 22 mg (yield 55%) of the title compound.

[0876] 1 H NMR (400 MHz, DMSO-d6) δ13.15 (s, 1H), 7.50 (s, 1H), 7.44 (d, J = 8.5 Hz, 1H), 7.35 - 7.27 (m, 1H), 4.12 (q, J = 7.1 Hz, 2H), 3.79 (t, J = 4.6 Hz, 4H), 3.29 - 3.23 (m, 4H), 1.37 (t, J = 7.3 Hz, 3H).

[0877]

[0878]

[0879] Example 33: Preparation of N-(1-ethyl-2-morpholino-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide (Compound 33)

[0880]

[0881] Step 33-1: Preparation of 1-ethyl-2-morpholino-1H-benzo[d]imidazol-5-amine (compound 33-1)

[0882]

[0883]

[0884] In the same manner as in step 5-3, 4-(1-ethyl-5-nitro-1H-benzo[d]imidazol-2-yl)morpholine (80 mg, 0.29 mmol) and ammonium chloride (124 mg, 2.32 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (129 mg, 2.32 mmol) was added to the reaction mixture to obtain 48 mg (yield 68%) of the title compound.

[0885] 1 H NMR (400 MHz, DMSO-d6) δ7.05 (d, J = 8.4 Hz, 1H), 6.64 (d, J = 2.0 Hz, 1H), 6.45 (dd, J = 8.4, 2.1 Hz, 1H), 4.69 (s, 2H), 3.96 (q, J = 7.2) Hz, 2H), 3.80 - 3.67 (m, 4H), 3.17 - 3.05 (m, 4H), 1.36 (s, 2H), 1.30 (t, J = 7.2 Hz, 3H).

[0886]

[0887] Step 33-2: Preparation of N-(1-ethyl-2-morpholino-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide (compound 33)

[0888]

[0889]

[0890] In the same manner as in Step 1-2, 1-ethyl-2-morpholino-1H-benzo[d]imidazol-5-amine (40 mg, 0.16 mmol), sodium nitrite (17 mg, 0.24 mmol) and 1.0 M aqueous hydrochloric acid solution (0.5 mL, 0.49 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (21 mg, 0.32 mmol) and aqueous sodium hydroxide solution to obtain 16 mg (yield 31%) of the title compound.

[0891] 1 H NMR (400 MHz, DMSO-d6) δ13.06 (s, 1H), 7.47 (s, 2H), 7.31 (d, J = 8.5 Hz, 1H), 4.10 (q, J = 7.2 Hz, 2H), 3.81 - 3.74 (m, 4H), 3.27 (s, 4H), 1.36 (t, J = 7.2 Hz, 3H).

[0892]

[0893] Example 34: Preparation of N-(2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide

[0894] Step 34-1: Preparation of 8-(6-nitro-1H-benzo[d]imidazol-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (Compound 34-1)

[0895]

[0896]

[0897] In the same manner as in Step 5-1, 332 mg (yield 80%) of the title compound was obtained from 2-chloro-6-nitro-1H-benzoimidazole (300 mg, 1.52 mmol), 3-oxa-8-azabicyclo[3.2.1]octane (344 mg, 3.04 mmol) and toluene.

[0898] 1 H NMR (400 MHz, DMSO-d6) δ12.31 - 12.00 (m, 1H), 8.06 - 7.86 (m, 2H), 7.33 (d, J = 8.8 Hz, 1H), 4.39 (s, 2H), 3.72 (d, J = 10.9 Hz, 2H), 3.61 (d, J = 10.9 Hz, 2H), 2.05 - 1.92 (m, 4H).

[0899]

[0900] Step 34-2:

[0901] 8-(1-Methyl-6-nitro-1H-benzo[d]imidazol-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (Compound 34-2)

[0902] 8-(1-Methyl-5-nitro-1H-benzo[d]imidazol-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (Compound 34-3)

[0903]

[0904]

[0905]

[0906] In the same manner as in Step 5-2, 118 mg (yield 37%) of the title compound (34-2) and 66 mg (yield 21%) of the title compound (34-3) were obtained from 8-(6-nitro-1H-benzo[d]imidazol-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (300 mg, 0.81 mmol), DBU (491 μL, 3.28 mmol), methyl iodide (204 μL, 3.28 mmol) and dimethylformamide.

[0907] Compound 34-2: 1 H NMR (400 MHz, Chloroform-d) δ8.17 - 8.06 (m, 2H), 7.50 (d, J = 8.6 Hz, 1H), 4.18 (s, 2H), 4.00 (d, J = 10.8 Hz, 2H), 3.75 - 3.70 (m, 5H), 2.14 (s, 4H).

[0908] Compound 34-3: 1 H NMR (400 MHz, Chloroform-d) δ8.42 (d, J = 2.2 Hz, 1H), 8.10 (dd, J = 8.8, 2.2 Hz, 1H), 7.18 (d, J = 8.8 Hz, 1H), 4.10 (s, 2H), 4.00 (d, J = 10.8 Hz, 2H), 3.76 - 3.68 (m, 5H), 2.20 - 2.07 (m, 4H).

[0909]

[0910]

[0911] Step 34-3: Preparation of 2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-6-amine (compound 34-4)

[0912]

[0913]

[0914] In the same manner as in step 5-3, 8-(1-methyl-6-nitro-1H-benzo[d]imidazol-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (200 mg, 0.69 mmol) and ammonium chloride (297 mg, 5.55 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (310 mg, 5.55 mmol) was added to the reaction mixture to obtain 170 mg (yield 97%) of the title compound.

[0915] 1 H NMR (400 MHz, DMSO-d6) δ7.03 (d, J = 8.4 Hz, 1H), 6.44 (d, J = 2.1 Hz, 1H), 6.38 (dd, J = 8.3, 2.1 Hz, 1H), 4.73 (s, 2H), 3.92 (s, 2H), 3.82 (d, J = 10.5 Hz, 2H), 3.60 - 3.55 (m, 2H), 3.49 (s, 3H), 2.02 - 1.81 (m, 4H).

[0916]

[0917] Step 34-4: Preparation of N-(2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide (compound 34)

[0918]

[0919]

[0920] In the same manner as in Step 1-2, 2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-6-amine (100 mg, 0.39 mmol), sodium nitrite (40 mg, 0.58 mmol) and 1.0 M aqueous hydrochloric acid solution (1.2 mL, 1.16 mmol) were dissolved in distilled water, and then the title compound (82 mg, 63% yield) was obtained through malononitrile (51 mg, 0.77 mmol) and aqueous sodium hydroxide solution.

[0921] 1H NMR (400 MHz, DMSO-d6) δ13.12 (s, 1H), 7.44 (d, J = 2.0 Hz, 1H), 7.37 (d, J = 8.5 Hz, 1H), 7.28 (dd, J = 8.6, 2.0 Hz, 1H), 4.19 (s, 2H), 3.85 (d, J = 10.8 Hz, 2H), 3.68 (s, 3H), 3.62 (dd, J = 11.0, 1.9 Hz, 2H), 2.06 - 1.85 (m, 4H).

[0922]

[0923] Example 35: Preparation of N-(2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide

[0924]

[0925] Step 35-1: Preparation of 2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-5-amine (Compound 35-1)

[0926]

[0927]

[0928] In the same manner as in step 5-3, 8-(1-methyl-5-nitro-1H-benzo[d]imidazol-2-yl)-3-oxa-8-azabicyclo[3.2.1]octane (130 mg, 0.45 mmol) and ammonium chloride (193 mg, 3.61 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (201 mg, 3.61 mmol) was added to the reaction mixture to obtain 37 mg (yield 32%) of the title compound.

[0929] 1H NMR (400 MHz, DMSO-d6) δ6.96 (d, J = 8.3 Hz, 1H), 6.57 (d, J = 2.0 Hz, 1H), 6.39 (dd, J = 8.3, 2.1 Hz, 1H), 4.54 (s, 2H), 3.99 (s, 2H), 3.82 (d, J = 10.5 Hz, 2H), 3.63 - 3.55 (m, 2H), 3.52 (s, 3H), 2.03 - 1.81 (m, 4H).

[0930]

[0931] Step 35-2 Preparation of N-(2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide (compound 35)

[0932]

[0933]

[0934] In the same manner as in Step 1-2, 2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-5-amine (20 mg, 0.08 mmol), sodium nitrite (8 mg, 0.12 mmol) and 1.0 M hydrochloric acid aqueous solution (0.2 mL, 0.23 mmol) were dissolved in distilled water, and then the title compound (15 mg, 57% yield) was obtained through malononitrile (10 mg, 0.15 mmol) and sodium hydroxide aqueous solution.

[0935] 1 H NMR (400 MHz, DMSO-d6) δ13.04 (s, 1H), 7.47 - 7.37 (m, 2H), 7.31 - 7.24 (m, 1H), 4.21 (s, 2H), 3.85 (d, J = 10.8 Hz, 2H), 3.67 (s, 3H), 3.62 (d, J = 10.8 Hz, 2H), 2.06 - 1.90 (m, 4H).

[0936]

[0937] Example 36: Preparation of N-(2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)-N-methylcarbonohydrazonoyl dicyanide

[0938]

[0939] Step 36-1: Preparation of N-(2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)-N-methylcarbonohydrazonoyl dicyanide

[0940]

[0941]

[0942] N-(2-(3-Oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (30 mg, 0.09 mmol) was dissolved in dimethylformamide under nitrogen, and potassium tert-butoxide (15 mg, 0.13 mmol) was added at room temperature, followed by iodomethane (28 μL, 0.44 mmol). The reaction mixture was stirred at 60 °C for 4 hours. Upon completion of the reaction, the mixture was extracted with distilled water and ethyl acetate, and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 8 mg (yield 26%) of the title compound.

[0943] 1 H NMR (400 MHz, DMSO-d6) δ8.00 (d, J = 2.4 Hz, 1H), 7.52 (d, J = 8.7 Hz, 1H), 7.42 (dd, J = 8.8, 2.4 Hz, 1H), 4.31 (s, 2H), 4.08 (s, 3H), 3.74 (d, J = 11.0 Hz, 2H), 3.62 (d, J = 10.8 Hz, 2H), 2.01 (s, 4H).

[0944]

[0945]

[0946] Example 37: Preparation of N-acetyl-N-(2-morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0947]

[0948] Step 37-1: Preparation of N-acetyl-N-(2-morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0949]

[0950]

[0951] N-(2-Morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (150 mg, 0.48 mmol) and potassium hydroxide (30 mg, 0.53 mmol) were dissolved in methanol and stirred at room temperature for 3 hours. After the reaction was completed, the solvent was concentrated under reduced pressure and solidified using ether. The resulting solid (170 mg, 0.48 mmol) and triethylamine (34 μL, 0.24 mmol) were dissolved in acetonitrile, and acetyl chloride (86 μL, 1.21 mmol) was added and stirred at room temperature for 4 hours. After the reaction was completed, extraction was performed using distilled water and ethyl acetate, and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, solidified using ether, and filtered. After concentrating the filtrate under reduced pressure again, the solid was solidified using hexane to obtain 64 mg (yield 17%) of the title compound.

[0952] 1 H NMR (400 MHz, Chloroform-d) δ7.63 (d, J = 8.6 Hz, 1H), 7.42 (d, J = 2.1 Hz, 1H), 7.07 (dd, J = 8.5, 2.2 Hz, 1H), 3.84 (t, J = 4.9 Hz, 4H), 3.67 (t, J = 4.9 Hz, 4H), 2.60 (s, 3H).

[0953]

[0954] Example 38: Preparation of N-(2-(morpholine-4-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0955]

[0956] Step 38-1: Preparation of phenyl(6-nitrobenzo[d]thiazol-2-yl)carbamate

[0957]

[0958]

[0959] 2-Amino-6-nitrobenzothiazole (500 mg, 2.56 mmol) and triethylamine (1.07 mL, 7.68 mmol) were dissolved in dichloromethane and dimethylformamide, and then phenyl chloroformate (386 μL, 3.07 mmol) was added. The reaction mixture was stirred at room temperature for 6 hours. Upon completion of the reaction, the resulting solid was filtered to obtain 802 mg (yield 99%) of the title compound.

[0960] 1 H NMR (400 MHz, DMSO-d6) δ13.07 (s, 1H), 9.07 (d, J = 2.4 Hz, 1H), 8.29 (dd, J = 9.1, 2.6 Hz, 1H), 7.90 (d, J = 8.9 Hz, 1H), 7.49 (t, J = 7.9) Hz, 2H), 7.35 - 7.28 (m, 3H).

[0961]

[0962] Step 38-2: Preparation of N-(6-nitrobenzo[d]thiazol-2-yl)morpholine-4-carboxamide

[0963]

[0964]

[0965] Phenyl(6-nitrobenzo[d]thiazol-2-yl)carbamate (200 mg, 0.63 mmol) was dissolved in dimethyl sulfoxide, and morpholine (277 μL, 3.17 mmol) was added. The reaction mixture was stirred at 50°C for 2 hours. Upon completion of the reaction, the mixture was extracted with aqueous sodium bicarbonate and ethyl acetate, and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 66 mg (yield 34%) of the title compound.

[0966] 1 H NMR (400 MHz, DMSO-d6) δ12.04 (s, 1H), 8.91 (d, J = 2.4 Hz, 1H), 8.23 ​​(dd, J = 8.9, 2.5 Hz, 1H), 7.69 (d, J = 8.9 Hz, 1H), 3.67 - 3.52 (m, 8H).

[0967]

[0968] Step 38-3: Preparation of N-(6-aminobenzo[d]thiazol-2-yl)morpholine-4-carboxamide

[0969]

[0970]

[0971] In the same manner as in step 5-3, N-(6-nitrobenzo[d]thiazol-2-yl)morpholine-4-carboxamide (50 mg, 0.16 mmol) and ammonium chloride (69 mg, 1.30 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (72 mg, 1.30 mmol) was added to the reaction mixture to obtain 34 mg (yield 74%) of the title compound.

[0972] 1 H NMR (400 MHz, DMSO-d6) δ10.91 (s, 1H), 7.22 (s, 1H), 6.91 (s, 1H), 6.63 (dd, J = 8.5, 2.2 Hz, 1H), 5.03 (s, 2H), 3.63 - 3.45 (m, 8H).

[0973]

[0974] Step 38-4: Preparation of N-(2-(morpholine-4-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (compound 38)

[0975]

[0976]

[0977] In the same manner as in Step 1-2, N-(6-aminobenzo[d]thiazol-2-yl)morpholine-4-carboxamide (30 mg, 0.11 mmol), sodium nitrite (11 mg, 0.16 mmol) and 1.0 M hydrochloric acid aqueous solution (0.3 mL, 0.32 mmol) were dissolved in distilled water, and then the resulting mixture was treated with malononitrile (14 mg, 0.22 mmol) and sodium hydroxide aqueous solution to obtain 35 mg (yield 92%) of the title compound.

[0978] 1 H NMR (400 MHz, Acetone-d6) δ12.00 (s, 1H), 7.98 (d, J = 2.2 Hz, 1H), 7.62 (d, J = 8.8 Hz, 1H), 7.55 (dd, J = 8.8, 2.2 Hz, 1H), 3.73 - 3.62 (m, 8H).

[0979]

[0980] Example 39: Preparation of N-(2-(2,6-dimethylmorpholine-4-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[0981] Step 39-1: Preparation of 2,6-dimethyl-N-(6-nitrobenzo[d]thiazol-2-yl)morpholine-4-carboxamide

[0982]

[0983]

[0984] In the same manner as in step 38-2, phenyl(6-nitrobenzo[d]thiazol-2-yl)carbamate (200 mg, 0.63 mmol) was dissolved in dimethyl sulfoxide solvent and then 2,6-dimethyl morpholine (391 μL, 3.17 mmol) was added to obtain 75 mg (yield 35%) of the title compound.

[0985] 1 H NMR (600 MHz, Chloroform-d) δ9.08 (s, 1H), 8.69 (d, J = 2.4 Hz, 1H), 8.31 - 8.23 ​​(m, 1H), 7.67 (d, J = 8.9 Hz, 1H), 3.93 (s, 2H), 3.63 (dt, J = 6.9, 3.8 Hz, 2H), 2.72 (t, J = 11.9 Hz, 2H), 1.21 (d, J = 6.2 Hz, 6H).

[0986]

[0987]

[0988] Step 39-2: Preparation of N-(6-aminobenzo[d]thiazol-2-yl)-2,6-dimethylmorpholine-4-carboxamide

[0989]

[0990]

[0991] In the same manner as in step 5-3, 2,6-dimethyl-N-(6-nitrobenzo[d]thiazol-2-yl)morpholine-4-carboxamide (50 mg, 0.15 mmol) and ammonium chloride (64 mg, 1.19 mmol) were dissolved in distilled water and tetrahydrofuran, and then iron (66 mg, 1.19 mmol) was added to the reaction mixture to obtain 44 mg (yield 97%) of the title compound.

[0992] 1H NMR (600 MHz, DMSO-d6) δ7.19 (s, 1H), 6.86 (d, J = 20.8 Hz, 1H), 6.60 (dd, J = 8.6, 2.2 Hz, 1H), 5.01 (s, 2H), 4.12 (s, 2H), 3.51 - 3.40 (m, 2H), 2.48 (s, 3H), 1.07 (d, J = 6.2 Hz, 6H).

[0993]

[0994] Step 39-3: Preparation of N-(2-(2,6-dimethylmorpholine-4-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (compound 39)

[0995]

[0996]

[0997] In the same manner as in Step 1-2, N-(6-aminobenzo[d]thiazol-2-yl)-2,6-dimethylmorpholine-4-carboxamide (40 mg, 0.13 mmol), sodium nitrite (13 mg, 0.20 mmol) and 1.0 M hydrochloric acid aqueous solution (0.4 mL, 0.39 mmol) were dissolved in distilled water, and then the resulting mixture was treated with malononitrile (17 mg, 0.26 mmol) and sodium hydroxide aqueous solution to obtain 18 mg (yield 35%) of the title compound.

[0998] 1 H NMR (400 MHz, Acetone-d6) δ11.95 (s, 1H), 10.14 (s, 1H), 7.98 (d, J = 2.2 Hz, 1H), 7.62 (d, J = 8.8 Hz, 1H), 7.55 (dd, J = 8.7, 2.2 Hz, 1H), 4.23 (dd, J = 13.4, 1.9 Hz, 2H), 3.62 (dqd, J = 14.9, 6.1, 2.3 Hz, 2H), 2.70 - 2.56 (m, 2H), 1.17 (d, J = 6.2 Hz, 6H).

[0999]

[1000] Example 40: Preparation of N-(2-(8-oxa-3-azabicyclo[3.2.1]octane-3-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[1001]

[1002] Step 40-1: Preparation of N-(6-nitrobenzo[d]thiazol-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane-3-carboxamide

[1003]

[1004]

[1005] In the same manner as in Step 38-2, phenyl(6-nitrobenzo[d]thiazol-2-yl)carbamate (200 mg, 0.63 mmol) was dissolved in dimethyl sulfoxide solvent, and then 8-oxa-3-azabicyclo[3.2.1]octane hydrochloride (474 ​​mg, 3.17 mmol) and triethylamine (442 μL, 3.17 mmol) were added to obtain 90 mg (yield 42%) of the title compound.

[1006] 1 H NMR (600 MHz, DMSO-d6) δ11.65 (s, 1H), 8.89 (s, 1H), 8.21 (dd, J = 8.9, 2.4 Hz, 1H), 7.68 (s, 1H), 4.34 (d, J = 2.6 Hz, 2H), 3.85 (s, 2H), 3.08 (d, J = 11.7 Hz, 2H), 1.83 - 1.62 (m, 4H).

[1007]

[1008] Step 40-2: Preparation of N-(6-nitrobenzo[d]thiazol-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane-3-carboxamide

[1009]

[1010]

[1011] In the same manner as in Step 5-3, N-(6-nitrobenzo[d]thiazol-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane-3-carboxamide (70 mg, 0.21 mmol) and ammonium chloride (90 mg, 1.67 mmol) were dissolved in distilled water and tetrahydrofuran, and iron (93 mg, 1.67 mmol) was added to the reaction mixture, and after removal of the solvent, it was used in the next step without purification.

[1012]

[1013]

[1014] Step 40-3: Preparation of N-(2-(8-oxa-3-azabicyclo[3.2.1]octane-3-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide (compound 40)

[1015]

[1016]

[1017] In the same manner as in Step 1-2, N-(6-nitrobenzo[d]thiazol-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane-3-carboxamide (50 mg, 0.16 mmol), sodium nitrite (17 mg, 0.25 mmol) and 1.0 M aqueous hydrochloric acid solution (0.5 mL, 0.49 mmol) were dissolved in distilled water, and then the resulting mixture was treated with malononitrile (22 mg, 0.33 mmol) and aqueous sodium hydroxide solution to obtain 43 mg (yield 69%) of the title compound.

[1018] 1 H NMR (600 MHz, DMSO-d6) δ13.12 (s, 1H), 11.18 (s, 1H), 7.96 (s, 1H), 7.63 (s, 1H), 7.49 (s, 1H), 4.33 (s, 2H), 3.81 (s, 2H), 3.06 (s, 2H), 1.84 - 1.73 (m, 3H).

[1019]

[1020] Example 41: Preparation of (R)-N-(2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 41)

[1021]

[1022] Step 41-1: Preparation of (R)-2-(3-fluoropyrrolidin-1-yl)-6-nitrobenzo[d]oxazole (Compound 41-1)

[1023]

[1024]

[1025] In the same manner as in step 3-2, 6-nitrobenzo[d]oxazole-2(3H)-thione (300 mg, 1.53 mmol), (R)-(-)-3-fluoropyrrolidine hydrochloride (576 mg, 4.59 mmol), triethylamine (1.07 mL, 7.64 mmol) and toluene were added and reacted at 180°C for 30 minutes using microwave irradiation, to obtain 253 mg (yield 66%) of the title compound.

[1026] 1 H NMR (400 MHz, DMSO-d6) δ8.32 (d, J = 2.2 Hz, 1H), 8.16 (dd, J = 8.7, 2.2 Hz, 1H), 7.40 (d, J = 8.7 Hz, 1H), 5.50 (d, J = 53 Hz, 1H), 3.93 - 3.80 (m, 3H), 3.74 - 3.67 (m, 1H), 2.38 - 2.16 (m, 2H).

[1027]

[1028] Step 41-2: Preparation of (R)-2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-amine (Compound 41-2)

[1029]

[1030]

[1031] In the same manner as in step 5-3, (R)-2-(3-fluoropyrrolidin-1-yl)-6-nitrobenzo[d]oxazole (200 mg, 0.79 mmol) and ammonium chloride (213 mg, 3.98 mmol) were dissolved in distilled water, tetrahydrofuran, and ethanol, and then iron (222 mg, 3.98 mmol) was added to the reaction mixture to obtain 114 mg (yield 65%) of the title compound.

[1032] 1 H NMR (400 MHz, DMSO-d6) δ6.96 (d, J = 8.3 Hz 1H), 6.65 (d, J = 2 Hz, 1H), 6.42 (dd, J = 8.3, 2 Hz, 1H), 5.43 (d, J = 53 Hz, 1H), 4.84 (s, 2H), 3.74 - 3.67 (m, 2H), 3.59 - 3.50 (m, 2H), 2.29 - 2.09 (m, 2H).

[1033]

[1034]

[1035] Step 41-3: Preparation of (R)-N-(2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 41)

[1036]

[1037]

[1038] In the same manner as in Step 1-2, (R)-2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-amine (100 mg, 0.45 mmol), sodium nitrite (77 mg, 0.90 mmol) and 1.0 M aqueous hydrochloric acid solution (1.36 mL, 1.36 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (60 mg, 0.90 mmol) and aqueous sodium hydroxide solution to obtain 80 mg (yield 59%) of the title compound.

[1039] 1H NMR (600 MHz, DMSO-d6) δ13.05 (s, 1H), 7.50 (d, J = 1.6 Hz 1H), 7.34 - 7.28 (m, 2H), 5.47 (d, J = 53 Hz, 1H), 3.85 - 3.73 (m, 2H), 3.66 - 3.59 (m, 2H), 2.31 - 2.13 (m, 2H).

[1040]

[1041] Example 42: Preparation of (S)-N-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 42)

[1042]

[1043] Step 42-1: Preparation of (S)-2-(3-fluoropiperidin-1-yl)-6-nitrobenzo[d]oxazole (Compound 42-1)

[1044]

[1045]

[1046] In the same manner as in step 3-2, 6-nitrobenzo[d]oxazole-2(3H)-thione (300 mg, 1.53 mmol), (S)-3-fluoropiperidine hydrochloride (640 mg, 4.59 mmol), triethylamine (639 μL, 4.59 mmol) and toluene were added and reacted at 180°C for 30 minutes using microwave irradiation, to obtain 238.9 mg (yield 59%) of the title compound.

[1047] 1H NMR (400 MHz, DMSO-d6) δ8.30 (d, J = 2.2 Hz, 1H), 8.15 (dd, J = 8.7, 2.3 Hz, 1H), 7.40 (d, J = 8.7 Hz, 1H), 4.94 (d, J = 48.3 Hz, 1H), 4.18 - 4.11 (m, J = 1H), 4.03 (dt, J = 13.2, 4.28 Hz, 1H), 3.69 (qd, J = 14.2, 2.0 Hz, 1H), 3.47 - 3.41 (m, 1H), 1.99 - 1.79 (m, 3H), 1.70 - 1.63 (m, 1H).

[1048]

[1049]

[1050] Step 42-2: Preparation of (S)-2-(3-fluoropyrrolphen-1-yl)benzo[d]oxazol-6-amine (compound 42-2)

[1051]

[1052]

[1053] In the same manner as in step 5-3, (S)-2-(3-fluoropyrrolphen-1-yl)-6-nitrobenzo[d]oxazole (200 mg, 0.79 mmol) and ammonium chloride (323 mg, 6.03 mmol) were dissolved in distilled water, tetrahydrofuran, and ethanol, and then iron (337 mg, 6.03 mmol) was added to the reaction mixture to obtain 153 mg (yield 87%) of the title compound.

[1054] 1H NMR (400 MHz, DMSO-d6) δ 6.94 (d, J = 8.3 Hz 1H), 6.61 (d, J = 2.0 Hz, 1H), 6.40 (dd, J = 8.2, 2.1 Hz, 1H), 4.88 - 4.76 (m, 3H), 3.89 - 3.82 (m, 1H), 3.75 (dt, J = 12.9, 4.5 1H), 3.55 (qd, J = 13.8, 2.4, 1H), 1.90 - 1.79 (m, 3H), 1.58 - 1.53 (m, 1H).

[1055]

[1056]

[1057] Step 42-3: Preparation of (S)-N-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 42)

[1058]

[1059]

[1060] In the same manner as in Step 1-2, (S)-2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-amine (120 mg, 0.51 mmol), sodium nitrite (52.8 mg, 0.77 mmol) and 1.0 M aqueous hydrochloric acid solution (3.1 mL, 3.06 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (67.4 mg, 1.02 mmol) and aqueous sodium hydroxide solution to obtain 81.1 mg (yield 51%) of the title compound.

[1061] 1 H NMR (600 MHz, DMSO-d6) δ13.08 (s, 1H), 7.50 (d, J = 1.8 Hz 1H), 7.34 - 7.27 (m, 2H), 4.89 (d, J = 47 Hz, 1H), 4.06 - 3.99 (m, 1H), 3.91 (dt, J = 13.2, 4.08, 1H), 3.62 (qd, J = 14, 2.0, 1H), 1.94 - 1.78 (m, 3H), 1.63 - 1.58 (m, 1H).

[1062]

[1063] Example 43: Preparation of (S)-N-(2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 43)

[1064]

[1065] Step 41-1: Preparation of (S)-2-(3-fluoropyrrolidin-1-yl)-6-nitrobenzo[d]oxazole (Compound 43-1)

[1066]

[1067]

[1068] In the same manner as in step 3-2, 6-nitrobenzo[d]oxazole-2(3H)-thione (600 mg, 3.09 mmol), (S)-(-)-3-fluoropyrrolidine hydrochloride (583 mg, 4.64 mmol), triethylamine (1.26 mL, 9.28 mmol) and toluene were added and reacted at 180°C for 30 minutes using microwave irradiation, to obtain 300 mg (yield 38.7%) of the title compound.

[1069] 1 H NMR (400 MHz, DMSO-d6) δ8.33 (d, J = 2.2 Hz, 1H), 8.17 (dd, J = 8.7, 2.3 Hz, 1H), 7.43 (d, J = 8.7 Hz, 1H), 5.51 (d, J = 53 Hz, 1H), 3.93 - 3.81 (m, 3H), 3.74 - 3.67 (m, 1H), 2.36 - 2.17 (m, 2H).

[1070]

[1071]

[1072] Step 43-2: Preparation of (S)-2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-amine (compound 43-2)

[1073]

[1074]

[1075] In the same manner as in step 5-3, (S)-2-(3-fluoropyrrolidin-1-yl)-6-nitrobenzo[d]oxazole (283 mg, 1.12 mmol) and ammonium chloride (301 mg, 5.63 mmol) were dissolved in distilled water, tetrahydrofuran, and ethanol, and then iron (315 mg, 5.63 mmol) was added to the reaction mixture to obtain 110 mg (yield 44%) of the title compound.

[1076] 1 H NMR (400 MHz, DMSO-d6) δ6.96 (d, J = 8.2 Hz 1H), 6.66 (d, J = 1.96 Hz, 1H), 6.42 (dd, J = 8.2, 2.0 Hz, 1H), 5.44 (d, J = 53 Hz, 1H), 4.85 (s, 2H), 3.78 - 3.68 (m, 2H), 3.59 - 3.55 (m, 1H), 2.29 - 2.09 (m, 2H).

[1077]

[1078]

[1079] Step 43-3: Preparation of (S)-N-(2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 43)

[1080]

[1081]

[1082] In the same manner as in Step 1-2, (S)-2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-amine (105 mg, 0.47 mmol), sodium nitrite (81 mg, 0.95 mmol) and 1.0 M aqueous hydrochloric acid solution (1.42 mL, 1.42 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (63 mg, 0.95 mmol) and aqueous sodium hydroxide solution to obtain 58 mg (yield 40.8%) of the title compound.

[1083] 1H NMR (600 MHz, DMSO-d6) δ13.08 (s, 1H), 7.53 (d, J = 1.7 Hz, 1H), 7.36 - 7.30 (m, 2H), 5.49 (d, J = 53 Hz, 1H), 3.87 - 3.74 (m, 3H), 3.68 - 3.61 (m, 1H), 2.33 - 2.14 (m, 2H).

[1084]

[1085] Example 44: Preparation of N-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 44)

[1086]

[1087] Step 44-1: Preparation of 2-(3-fluoropiperidin-1-yl)-6-nitrobenzo[d]oxazole (compound 44-1)

[1088]

[1089]

[1090] In the same manner as in step 3-2, 6-nitrobenzo[d]oxazole-2(3H)-thione (500 mg, 2.55 mmol), 3-fluoropiperidine hydrochloride (712 mg, 5.10 mmol), triethylamine (1.4 mL, 10.2 mmol) and toluene were added and reacted at 180°C for 30 minutes using microwave irradiation, to obtain 363 mg (yield 54%) of the title compound.

[1091] 1 H NMR (400 MHz, DMSO-d6) δ8.30 (S, 1H), 8.16 (d, J = 8.7 Hz, 1H), 7.40 (d, J = 8.7 Hz, 1H), 4.95 (d, J = 46.8 Hz, 1H), 4.16 (t, J = 11.92, 1H), 4.04 (d, J = 13 Hz, 1H), 3.69 (dd, J = 32.9, 13.9 Hz, 1H), 3.47 - 3.41 (m, 1H), 1.96 - 1.67 (m, 4H).

[1092]

[1093] Step 44-2: Preparation of 2-(3-fluoropyrrolphen-1-yl)benzo[d]oxazol-6-amine (compound 44-2)

[1094]

[1095]

[1096] In the same manner as in step 5-3, 2-(3-fluoropyrrolphen-1-yl)-6-nitrobenzo[d]oxazole (300 mg, 1.13 mmol) and ammonium chloride (484 mg, 9.05 mmol) were dissolved in distilled water, tetrahydrofuran, and ethanol, and then iron (505 mg, 9.05 mmol) was added to the reaction mixture to obtain 231 mg (yield 87%) of the title compound.

[1097] 1 H NMR (400 MHz, DMSO-d6) δ6.95 (d, J = 8.2 Hz 1H), 6.62 (d, J = 2.0 Hz, 1H), 6.41 (dd, J = 8.3, 2.1 Hz, 1H), 4.89 - 4.76 (m, 3H), 3.90 - 3.83 (m, 1H), 3.78 - 3.73 (m, 1H), 3.56 (qd, J = 13.8, 2.4, 1H), 1.91 - 1.79 (m, 3H), 1.59 - 1.54 (m, 1H).

[1098]

[1099]

[1100] Step 44-3: Preparation of N-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 44)

[1101]

[1102]

[1103] In the same manner as in Step 1-2, 2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-amine (200 mg, 0.85 mmol), sodium nitrite (88 mg, 1.28 mmol) and 1.0 M aqueous hydrochloric acid solution (5.1 mL, 5.1 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (112 mg, 1.70 mmol) and aqueous sodium hydroxide solution to obtain 111 mg (yield 42%) of the title compound.

[1104] 1 H NMR (600 MHz, DMSO-d6) δ13.06 (s, 1H), 7.50 (d, J = 1.8 Hz 1H), 7.34 - 7.27 (m, 2H), 4.89 (d, J = 47 Hz, 1H), 4.06 - 3.99 (m, 1H), 3.93 - 3.89 (m, 1H), 3.62 (qd, J = 14, 2.0, 1H), 1.94 - 1.81 (m, 3H), 1.63 - 1.58 (m, 1H).

[1105]

[1106] Example 45: Preparation of N-(2-(pyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 45)

[1107]

[1108] Step 45-1: Preparation of 2-(pyrrolidin-1-yl)-6-nitrobenzo[d]oxazole (compound 45-1)

[1109]

[1110]

[1111] In the same manner as in step 3-2, 6-nitrobenzo[d]oxazole-2(3H)-thione (300 mg, 1.53 mmol), pyrrolidine (255 μL, 3.06 mmol), triethylamine (852 μL, 6.12 mmol) and toluene were added and refluxed at 120°C for 2 hours to obtain 119 mg (yield 33%) of the title compound.

[1112] 1 H NMR (400 MHz, DMSO-d6) δ8.28 (S, 1H), 8.14 (dd, J = 8.7 Hz, 2.28, 1H), 7.37 (d, J = 8.7 Hz, 1H), 3.62 (t, J = 6.64 Hz, 4H), 2.01 - 1.98 (m, 4H).

[1113]

[1114]

[1115] Step 45-2: Preparation of 2-(pyrrolidin-1-yl)benzo[d]oxazol-6-amine (compound 45-2)

[1116]

[1117]

[1118] In the same manner as in step 5-3, 2-(pyrrolidin-1-yl)-6-nitrobenzo[d]oxazole (100 mg, 0.43 mmol) and ammonium chloride (183 mg, 3.43 mmol) were dissolved in distilled water, tetrahydrofuran, and ethanol, and then iron (192 mg, 3.43 mmol) was added to the reaction mixture to obtain 72 mg (yield 82%) of the title compound.

[1119] 1 H NMR (400 MHz, CDCl3-d1) δ7.44 (d, J = 8.2 Hz 1H), 6.69 (d, J = 2.1 Hz, 1H), 6.54 (dd, J = 8.2, 2.1 Hz, 1H), 3.63 - 3.60 (m, 6H), 2.04 - 2.00 (m, 4H).

[1120]

[1121] Step 45-3: Preparation of N-(2-(pyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 45)

[1122]

[1123]

[1124] In the same manner as in Step 1-2, 2-(pyrrolidin-1-yl)benzo[d]oxazol-6-amine (60 mg, 0.30 mmol), sodium nitrite (31 mg, 0.44 mmol) and 1.0 M hydrochloric acid aqueous solution (1.8 mL, 1.77 mmol) were dissolved in distilled water, and then the title compound (56 mg, 67% yield) was obtained through malononitrile (39 mg, 0.59 mmol) and sodium hydroxide aqueous solution.

[1125] 1 H NMR (600 MHz, DMSO-d6) δ13.09 (s, 1H), 7.50 (d, J = 1.8 Hz 1H), 7.34 - 7.26 (m, 2H), 3.58 - 3.55 (m, 4H), 2.00 - 1.96 (m, 4H).

[1126]

[1127] Example 46: Preparation of N-(2-(piperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 46)

[1128]

[1129] Step 46-1: Preparation of 2-(piperidin-1-yl)-6-nitrobenzo[d]oxazole (compound 46-1)

[1130]

[1131]

[1132] In the same manner as in step 3-2, 6-nitrobenzo[d]oxazole-2(3H)-thione (300 mg, 1.53 mmol), piperidine (302 μL, 3.06 mmol), triethylamine (852 μL, 6.12 mmol) and toluene were added and refluxed at 170 °C for 19 hours to obtain 233 mg of the title compound, which was then subjected to the next reaction without separation.

[1133]

[1134]

[1135] Step 46-2: Preparation of 2-(piperidin-1-yl)benzo[d]oxazol-6-amine (compound 46-2)

[1136]

[1137]

[1138] In the same manner as in step 5-3, 2-(piperidin-1-yl)-6-nitrobenzo[d]oxazole (200 mg, 0.81 mmol) and ammonium chloride (346 mg, 6.47 mmol) were dissolved in distilled water, tetrahydrofuran, and ethanol, and then iron (361 mg, 6.47 mmol) was added to the reaction mixture to obtain 126 mg (yield 72%) of the title compound.

[1139] 1 H NMR (400 MHz, DMSO-d6) δ6.94 (d, J = 8.2 Hz 1H), 6.62 (d, J = 2.0 Hz, 1H), 6.41 (dd, J = 8.2, 2.1 Hz, 1H), 4.85 (s, 2H), 3.50 - 3.49 (m, 4H), 1.66 - 1.55 (m, 6H).

[1140]

[1141]

[1142] Step 46-3: Preparation of N-(2-(piperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (compound 46)

[1143]

[1144]

[1145] In the same manner as in Step 1-2, 2-(piperidin-1-yl)benzo[d]oxazol-6-amine (110 mg, 0.51 mmol), sodium nitrite (52 mg, 0.76 mmol) and 1.0 M aqueous hydrochloric acid solution (3.04 mL, 3.04 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (67 mg, 1.01 mmol) and aqueous sodium hydroxide solution to obtain 56 mg (yield 37%) of the title compound.

[1146] 1 H NMR (600 MHz, DMSO-d6) δ13.0 (s, 1H), 7.49 (d, J = 1.8 Hz 1H), 7.34 - 7.26 (m, 2H), 3.62 - 3.61 (m, 4H), 1.66 - 1.58 (m, 6H).

[1147]

[1148] Example 47: Preparation of (R)-N-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 47)

[1149] Step 47-1: Preparation of (R)-2-(3-fluoropiperidin-1-yl)-6-nitrobenzo[d]oxazole (Compound 47-1)

[1150]

[1151]

[1152] In the same manner as in step 3-2, 6-nitrobenzo[d]oxazole-2(3H)-thione (500 mg, 2.55 mmol), (R)-3-fluoropiperidine hydrochloride (712 mg, 5.10 mmol), triethylamine (1.4 mL, 10.2 mmol) and toluene were added and reacted at 160°C for 30 minutes using microwave irradiation, to obtain 402.7 mg (yield 61.6%) of the title compound.

[1153] 1 H NMR (400 MHz, DMSO-d6) δ8.31 (d, J = 2.24 Hz 1H), 8.16 (dd, J = 8.68, 2.28 Hz, 1H), 7.41 (d, J = 8.72 Hz, 1H), 5.02 - 4.88 (m, 1H), 4.20 - 4.03 (m, 2H), 3.76 - 3.63 (m, 1H), 3.48 - 3.41 (m, 1H), 2.00 - 1.81 (m, 3H), 1.69 - 1.65 (m, 1H).

[1154]

[1155]

[1156] Step 47-2: Preparation of (R)-2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-amine (Compound 47-2)

[1157]

[1158]

[1159] In the same manner as in step 5-3, (R)-2-(3-fluoropiperidin-1-yl)-6-nitrobenzo[d]oxazole (350 mg, 1.32 mmol) and ammonium chloride (5+65 mg, 10.6 mmol) were dissolved in distilled water, tetrahydrofuran, and ethanol, and then iron (590 mg, 10.6 mmol) was added to the reaction mixture to obtain 254.9 mg (yield 82%) of the title compound.

[1160] 1 H NMR (400 MHz, DMSO-d6) δ6.95 (d, J = 8.2 Hz 1H), 6.62 (d, J = 1.9 Hz, 1H), 6.41 (dd, J = 8.5, 2.1 Hz, 1H), 4.90 - 4.77 (m, 3H), 3.91 - 3.74 (m, 2H), 3.62 - 3.50 (m, 1H), 1.91 - 1.79 (m, 3H), 1.60 - 1.55 (m, 1H).

[1161]

[1162]

[1163] Step 47-3: Preparation of (R)-N-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide (Compound 47)

[1164]

[1165]

[1166] In the same manner as in Step 1-2, (R)-2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-amine (200 mg, 0.85 mmol), sodium nitrite (90 mg, 1.3 mmol) and 1.0 M aqueous hydrochloric acid solution (5.10 mL, 5.10 mmol) were dissolved in distilled water, and then the mixture was treated with malononitrile (112 mg, 1.7 mmol) and aqueous sodium hydroxide solution to obtain 173 mg (yield 65.2%) of the title compound.

[1167] 1 H NMR (400 MHz, DMSO-d6) δ13.1 (s, 1H), 7.51 (d, J = 1.8 Hz 1H), 7.35 - 7.28 (m, 2H), 4.96 - 4.84 (m, 1H), 4.07 - 3.91 (m, 2H), 3.69 - 3.57 (m, 1H), 3.42 - 3.39 (m, 1H), 1.95 - 1.80 (m, 3H), 1.65 - 1.61 (m, 1H).

[1168]

[1169] Example 48: Preparation of N-(2-(morpholine-4-carbonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)carbonohydrazonoyl dicyanide (compound 48)

[1170] Step 48-2: Preparation of morpholino(1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methanone (Compound 48-2)

[1171]

[1172]

[1173] N,N-diisopropylamine (DIPEA, 3.50 mL, 19.8 mmol) and HATU (Hexafluorophosphate Azabenzotriazole Tetramethyl Uronium, 4.90 g, 12.9 mmol) were added to 1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine-2-carboxylic acid (3 g, 9.90 mmol) under nitrogen, and the mixture was stirred at room temperature for 20 minutes. Then, morpholine (1.70 mL, 19.8 mmol) was added, and the mixture was stirred at room temperature for 5 hours to obtain 3.00 g (yield 83.11%) of the title compound.

[1174] 1 H NMR (400 MHz, DMSO-d6) δ8.43 (d, J = 4.7 Hz, 1H), 8.23 ​​(d, J = 8.0 Hz, 2H), 8.06 (d, J = 7.8 Hz, 1H), 7.73 (t, J = 7.3 Hz, 1H), 7.64 (t, J = 7.6 Hz, 2H), 7.34 (dd, J = 7.8 Hz, 4.8 Hz, 1H), 6.93 (s, 1H), 3.70 (s, 4H), 3.65 3.64 (m, 2H), 3.49 (m, 2H).

[1175]

[1176]

[1177] Step 48-3: Preparation of morpholino(5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methanone (Compound 48-3)

[1178]

[1179]

[1180] Morpholino(1-phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methanone (2 g, 5.40 mmol) and tetrabutylammonium nitrate (2.1 g, 7.0 mmol) were dissolved in dichloromethane under nitrogen, and the mixture was stirred at 0°C for 1.5 h. Then, trifluoroacetic anhydride (TFAA, 978 μL, 7.0 mmol) was added, and the mixture was stirred at 0°C for 10 min and then at room temperature for an additional 18.5 h. After the reaction was completed, the residue was purified by column chromatography to obtain 1.6 g (yield 73%) of the title compound.

[1181] 1 H NMR (400 MHz, DMSO-d6) δ9.26 (d, J = 2.4 Hz, 1H), 8.97 (d, J = 2.5 Hz, 1H), 8.28 (d, J = 7.7 Hz, 2H), 7.80 (t, J = 7.4 Hz, 1H), 7.70 (t, J = 7.7 Hz, 2H), 7.12 (s, 1H), 3.72 (s, 4H), 3.67 3.66 (m, 2H), 3.53 (m, 2H).

[1182]

[1183]

[1184] Step 48-4: Preparation of morpholino(5-nitro-1H-pyrrolo[2,3-b]pyridin-2-yl)methanone (compound 48-4)

[1185]

[1186]

[1187] Morpholino(5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methanone (1.4 g, 3.40 mmol) and solid potassium tert-butoxide (1.1 g, 10 mmol) were dissolved in dioxane under nitrogen and stirred at 80°C for 13 hours to obtain 325 mg (yield 35%) of the title compound.

[1188] 1H NMR (400 MHz, DMSO-d6) δ13.0 (s, 1H), 9.18 (d, J = 2.2 Hz, 1H), 8.96 (d, J = 1.8 Hz, 1H), 7.03 (s, 1H), 3.68 (d, J = 11.9 Hz, 8H).

[1189]

[1190]

[1191] Step 48-5: Preparation of tert-butyl 2-(morpholine-4-carbonyl)-5-nitro-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (compound 48-5)

[1192]

[1193]

[1194] Morpholino(5-nitro-1H-pyrrolo[2,3-b]pyridin-2-yl)methanone (300 mg, 1.09 mmol), di-tert-butyl dicarbonate (568 mg, 2.60 mmol), and 4-dimethylaminopyridine (26 mg, 0.2 mmol) were dissolved in tetrahydrofuran under nitrogen, and the mixture was stirred at room temperature for 22.5 hours. 186.7 mg (yield 46%) of the title compound was obtained after purification by column chromatography.

[1195] 1 H NMR (400 MHz, DMSO-d6) δ9.28 (d, J = 2.4 Hz, 1H), 8.99 (d, J = 2.4 Hz, 1H), 7.08 (s, 1H), 3.70-3.52 (m, 8H), 1.56 (s, 9H).

[1196]

[1197]

[1198] Step 48-6: Preparation of tert-butyl 5-amino-2-(morpholine-4-carbonyl)-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (compound 48-6)

[1199]

[1200]

[1201] tert-Butyl 2-(morpholine-4-carbonyl)-5-nitro-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (150 mg, 0.40 mmol) was dissolved in ethyl acetate under nitrogen, and 10% carbon-activated palladium catalyst (47 mg, 0.065 mmol) was added. The mixture was stirred at room temperature for 10 hours under hydrogen gas to obtain 120.2 mg (yield 86%) of the title compound.

[1202] 1 H NMR (400 MHz, DMSO-d6) δ7.86 (d, J = 2.4 Hz, 1H), 7.12 (d, J = 2.4 Hz, 1H), 6.60 (s, 1H), 5.16 (s, 2H), 3.67-3.44 (m, 8H), 1.51 (s, 9H).

[1203]

[1204]

[1205] Step 48-7: Preparation of tert-butyl 5-(2-(dicyano-methylene)hydrazinyl)-2-(morpholine-4-carbonyl)-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (compound 48-7)

[1206]

[1207]

[1208] Tert-Butyl 5-amino-2-(morpholine-4-carbonyl)-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (50 mg, 0.14 mmol) and sodium nitrite (15 mg, 0.22 mmol) were dissolved in ethanol under nitrogen, and then 1.0 M aqueous hydrochloric acid solution (0.4 mL, 0.36 mmol) was added at 0°C. The reaction mixture was stirred at 0°C for 1 h to form a diazonium salt. After the diazonium salt was formed, malononitrile (14 mg, 0.22 mmol) was added and stirred at room temperature for 3.5 h to obtain 46.6 mg (yield 79%) of the title compound.

[1209] 1 H NMR (400 MHz, DMSO-d6) δ8.57 (s, 1H), 8.05 (s, 1H), 6.87 (s, 1H), 3.69-3.49 (m, 8H), 1.55 (s, 9H).

[1210]

[1211]

[1212] Step 48-8: Preparation of N-(2-(morpholine-4-carbonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)carbonohydrazonoyl dicyanide (compound 48)

[1213]

[1214]

[1215] tert-Butyl 5-(2-(dicyano-methylene)hydrazinyl)-2-(morpholine-4-carbonyl)-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (40 mg, 0.09 mmol) was dissolved in dichloromethane under nitrogen, and trifluoroacetic acid (145 μL, 1.89 mmol) was added. The reaction mixture was stirred at room temperature for 1.5 hours to obtain 17.3 mg (yield 59.4%) of the title compound.

[1216] 1 H NMR (400 MHz, DMSO-d6) δ13.2 (s, 1H), 12.2 (s, 1H), 8.51 (s, 1H), 8.04 (s, 1H), 6.81 (s, 1H) 3.70-3.66 (m, 8H)

[1217]

[1218] Example 49: Preparation of N-(2-(morpholinooxazolo[4,5-b]pyridin-6-yl)carbonohydrazonoyl dicyanide (compound 49)

[1219]

[1220] Step 49-1: Preparation of oxazolno[4,5-b]pyridine-2(3H)-thione (compound 49-1)

[1221]

[1222] 2-Aminopyridin-3-ol (5 g, 45.4 mmol) and potassium hydroxide (7.6 g, 136 mmol) were dissolved in ethanol, and then carbon disulfide (54.6 mL, 908 mmol) was added. The reaction mixture was stirred at 75°C for 24 hours to obtain 5.026 g (yield 73%) of the title compound.

[1223] 1H NMR (400 MHz, DMSO-d6) δ14.49 (s, 1H), 8.24 (d, J = 5.2 Hz, 1H), 7.89 (dd, J = 8.1, 1.5 Hz, 1H), 7.29 (dd, J = 7.9, 5.1 Hz, 1H).

[1224]

[1225]

[1226] Step 49-2: Preparation of 2-(methyltino)oxazolno[4,5-b]pyridine (Compound 49-2)

[1227]

[1228] Oxazolno[4,5-b]pyridine-2(3H)-thione (4.8 g, 32 mmol) and potassium carbonate (6.1 g, 44.8 mmol) were dissolved in ethyl acetate, and methyl iodide (5.0 mL, 78 mmol) was added. The reaction mixture was stirred at room temperature for 12.5 hours to obtain 4.533 g (yield 85%) of the title compound.

[1229] 1H NMR (400 MHz, DMSO-d6) δ8.44 (dd, J = 4.9, 1.5 Hz, 1H), 8.09 (dd, J = 8.2, 1.5 Hz, 1H), 7.35 (ddd, J = 8.4, 4.9, 1.3 Hz, 1H), 2.81 (d, J = 1.2 Hz, 3H).

[1230]

[1231] Step 49-3: Preparation of 2-morpholinooxazolno[4,5-b]pyridine (compound 49-3)

[1232]

[1233]

[1234] 2-(Methyltino)oxazolno[4,5-b]pyridine (4.5 g, 27.1 mmol) was dissolved in tetrahydrofuran, and morpholine (11.6 mL, 135 mmol) was added. The reaction mixture was stirred at 80°C for 13 hours to obtain 3.910 g (yield 70.3%) of the title compound.

[1235] 1H NMR (400 MHz, DMSO-d6) δ8.15 (dd, J = 5.3, 1.5 Hz, 1H), 7.79 - 7.72 (m, 1H), 7.02 (dd, J = 8.1, 5.4 Hz, 1H), 3.74 (t, J = 4.7 Hz, 4H), 3.66 (dd, J = 5.7, 3.8 Hz, 4H).

[1236]

[1237]

[1238] Step 49-4: Preparation of 2-morpholino-6-nitrooxazolo[4,5-b]pyridine (compound 49-4)

[1239]

[1240]

[1241] 2-Morpholinoxazolno[4,5-b]pyridine (3.6 g, 5.80 mmol) and tetrabutylammonium nitrate (2.6 g, 8.70 mmol) were dissolved in dichloromethane under nitrogen and stirred at room temperature for 3 hours. Then, trifluoroacetic anhydride (TFAA, 1.6 mL, 11.6 mmol) was added and stirred at 0°C for 10 minutes and then at room temperature for an additional 31 hours. After the reaction was completed, the residue was purified by column chromatography to obtain 620 mg (yield 14%) of the title compound.

[1242] 1H NMR (400 MHz, DMSO-d6) δ9.11 (d, J = 2.4 Hz, 1H), 8.56 (d, J = 2.3 Hz, 1H), 3.76 (s, 8H).

[1243]

[1244]

[1245] Step 49-5: Preparation of 2-morpholinooxazolo[4,5-b]pyridin-6-amine (compound 49-5)

[1246]

[1247]

[1248] 2-Morpholino-6-nitrooxazolo[4,5-b]pyridine (620 mg, 2.50 mmol) was dissolved in ethyl acetate, and 10% carbon-activated palladium catalyst (590 mg, 0.5 mmol) was added. The mixture was stirred at room temperature for 7 hours under hydrogen gas. After the reaction was complete, the residue was purified by column chromatography to obtain 620 mg (yield 20%) of the title compound.

[1249] 1H NMR (400 MHz, DMSO-d6) δ7.58 (d,J= 2.3 Hz, 1H), 7.01 (d,J= 2.3 Hz, 1H), 5.05 (s, 2H), 3.72 (t,J= 4.9 Hz, 4H), 3.55 (t,J= 4.9 Hz, 4H).

[1250]

[1251] Step 49-6: Preparation of N-(2-(morpholinooxazolo[4,5-b]pyridin-6-yl)carbonohydrazonoyl dicyanide (compound 49-6)

[1252]

[1253]

[1254] 2-Morpholinoxazolo[4,5-b]pyridin-6-amine (50 mg, 0.23 mmol) and sodium nitrite (24 mg, 0.34 mmol) were dissolved in ethanol, and 1.0 M aqueous hydrochloric acid solution (0.7 mL, 0.68 mmol) was added at room temperature. The reaction mixture was stirred at room temperature for 1 hour to form a diazonium salt. After the diazonium salt was formed, malononitrile (22.5 mg, 0.34 mmol) was added, stirred at room temperature for 1 hour, and purified by column chromatography to obtain 13.2 mg (yield 19%) of the title compound.

[1255] 1H NMR (400 MHz, DMSO-d6) δ13.6 (s, 1H), 8.32 (s, 1H), 7.83 (s, 1H), 3.75 (t, J = 4.6 Hz, 4H), 3.67 (d, J = 4.7 Hz, 4H).

[1256]

[1257] Comparative Examples 1 to 5: Preparation of benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide containing heteroaryl or acyclic substituents

[1258]

[1259] The following six compounds were synthesized according to the method disclosed in the applicant's prior patent, Patent Application No. 10-2020-0039398, and used as comparative compounds 1 to 6.

[1260] Comparative compound 1: N-(2-methylbenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide

[1261] Comparative compound 2: N-(2-phenylbenzo[d]thiazol-6-yl) carbonohydrazonoyl dicyanide

[1262] Comparative compound 3: N-(2-(4-fluorophenyl)benzo[d]thiazol-6-yl) carbonohydrazonoyl dicyanide

[1263] Comparative compound 4: N-(2-(4-(dimethylamino)phenyl)benzo[d]thiazol-6-yl) carbonohydrazonoyl dicyanide

[1264] Comparative compound 5: N-(2-(thiophen-2-yl)benzo[d]thiazol-6-yl) carbonohydrazonoyl dicyanide

[1265] Comparative compound 6: N-(2-(6-methylpyridin-3-yl) benzo[d]thiazol-6-yl) carbonohydrazonoyl dicyanide

[1266] Comparative compound 1 Comparative compound 2 Comparative compound 3 Comparative compound 4 Comparative compound 5 Comparative compound 6

[1267]

[1268] [Example]

[1269] Meanwhile, the novel compound represented by Chemical Formula 1 according to the present invention can be formulated into various forms depending on the purpose. The following are examples of several formulation methods containing the compound represented by Chemical Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

[1270]

[1271] Preparation Example 1: Preparation of tablets by direct pressurization

[1272]

[1273] 5.0 mg of the active ingredient manufactured according to Examples 1 to 46 above was sieved, and then mixed with 14.1 mg of lactose, 0.8 mg of crospovidone USNF, and 0.1 mg of magnesium stearate, and pressed to manufacture tablets.

[1274]

[1275] Preparation Example 2: Preparation of tablets by wet granulation

[1276]

[1277] 5.0 mg of the active ingredient prepared according to Examples 1 to 46 above was sieved and mixed with 16.0 mg of lactose and 4.0 mg of starch. 0.3 mg of polysorbate 80 was dissolved in pure water, and an appropriate amount of this solution was added to the mixture to form fine particles. After drying, the fine particles were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate, and the fine particles were pressed to form tablets.

[1278]

[1279] Formulation Example 3: Preparation of powder and capsules

[1280]

[1281] 5.0 mg of the active ingredient prepared according to Examples 1 to 46 above was sieved and mixed with 14.8 mg of lactose, 10.0 mg of polyvinylpyrrolidone, and 0.2 mg of magnesium stearate. The mixture was filled into hard No. 5 gelatin capsules using an appropriate device to prepare capsules.

[1282]

[1283] Preparation Example 4: Preparation of Injection

[1284]

[1285] An injection was prepared by containing 100 mg of the active ingredient manufactured according to Examples 1 to 46, and further including 180 mg of mannitol, 26 mg of Na2HPO412H2O, and 2974 mg of distilled water.

[1286]

[1287] Experimental Example 1: Screening of Tau Aggregation Inhibitors Using the Tau-BiFC Cell Model

[1288] To screen for novel tau aggregation inhibitors, we utilized a tau-BiFC cell model, which facilitates observation of tau oligomer formation in living cells. Tau-BiFC cells were seeded in 384-well plates, and the following day, the cells were treated with Forskolin (at a concentration of 30 μM), a compound that activates tau phosphorylation enzyme PKA and induces tau aggregation, along with the compounds prepared according to Examples 1 to 46 at concentrations of 1, 3, and 10 μM, respectively. After 48 hours, the nuclei in the cells were stained with Hoechst (at a concentration of 2 μg / mL), and the BiFC fluorescence intensity was automatically quantified using Operetta (Perkin Elmer), and the stained nuclei in each well were counted from the entire plate. The group treated only with Forskolin, which induces tau aggregation, was set as a reference point with 100% tau aggregation, and the effect was confirmed by the formula "BiFC fluorescence intensity by the compound synthesized according to the example of the present invention / (fluorescence intensity of the control group treated only with Forskolin, which induces tau aggregation - fluorescence intensity of the untreated control group) × 100". In addition, the degree of cytotoxicity induced by the newly synthesized compound was also calculated by the formula "(number of stained nuclei in the group treated with the compound / number of stained nuclei in the group treated with Forskolin) × 100", using the group treated only with Forskolin as a reference with 100% viability. From the treatment results, a substance that inhibits intracellular tau aggregation was selected from a series of candidates based on 70% or more tau aggregation inhibition and 100% cell viability at a compound treatment concentration of 10 μM.

[1289]

[1290] Experimental Example 2: Confirmation of the concentration-dependent tau aggregation inhibition effect of a novel compound.

[1291] To confirm the tau aggregation inhibitory effect according to the concentration of the compounds selected according to the above Experimental Example 1, the selected compounds were treated with tau-BiFC cells at concentrations of 0.03, 0.1, 0.3, 1, 3, 10, and 30 μM, respectively, together with Forskolin (treatment concentration 30 μM), a tau aggregation inducer. After 48 hours, the cell images were checked to analyze the tau aggregation reaction and the degree of cytotoxicity. IC of the compounds 50 Toxicity was analyzed using nonlinear regression analysis using Prism software (GraphPad). The results for representative compounds are shown in Table 3: Tau aggregation inhibition effect and cell viability of novel compounds.

[1292] Tau aggregation inhibitory effect (Tau-BiFC cell line) Compound number IC 50 (nM) Cell viability (% @10μM) Compound number IC 50 (nM) Cell viability (% @10 μM) 112 100 24 100 320 100 45 6 100 532 100 62 4 10 47 6 4 100 832 10 19 36 100 102 4 10 9 1132 100 1232 10 2132 0 9 144 8 100 154 8 100 164 9 100 178 8 100 1812 8 100 19 45 3100 20 119 62 348 9 22 4 168 9 5 25 36 9426109927449928169929369730609931881003212983349734121003512100361369737248538689739449840316984126010042201004328100441210045498461610047117100Comparative compound 1500100Comparative compound 22,80065Comparative compound 33,20050Comparative compound 43,10080Comparative compound 57,30070

[1293]

[1294] Experimental Example 3: Screening of Tdp-43 aggregation inhibitors using the Tau-BiFC cell model.

[1295] To screen for novel Tdp43 aggregation inhibitors, a Tdp43-BiFC cell model was used, which facilitates observation of Tdp43 aggregate formation in living cells. Specifically, Tdp43-BiFC cells were seeded in 384-well plates, and the following day, the cells were treated with Forskolin (treatment concentration: 30 μM), a compound that induces Tdp43 aggregation by activating the phosphorylation enzyme PKA, along with the compounds prepared according to Examples 1 to 46 at concentrations of 3 and 10 μM, respectively. After 48 hours, the nuclei in the cells were stained with Hoechst (treatment concentration: 1 μg / mL), and the BiFC fluorescence intensity was automatically quantified using Operetta (PerkinElmer), and the stained nuclei in each well from the entire plate were counted. The group treated only with Forskolin, which induces Tdp-43 aggregation, was set as a reference point with 100% Tdp-43 aggregation, and the effect was confirmed by the formula, "(BiFC fluorescence intensity by the compound synthesized according to the example of the present invention - fluorescence intensity of the untreated control group) / (fluorescence intensity of the control group treated only with Forskolin, which induces Tdp-43 aggregation - fluorescence intensity of the untreated control group) × 100." In addition, the degree of cytotoxicity induced by the newly synthesized compound was also calculated by the formula, "(number of stained nuclei in the group treated with compound / number of stained nuclei in the group treated with Forskolin) × 100," using the group treated only with Forskolin as a reference with 100% viability. From the treatment results, a substance that inhibits intracellular Tdp-43 aggregation was selected from a series of candidates based on 80% or more inhibition of Tdp-43 aggregation and 60% cell viability at a compound treatment concentration of 3 μM.

[1296]

[1297] Experimental Example 4: Confirmation of the concentration-dependent Tdp-43 aggregation inhibition effect of a novel compound.

[1298] Furthermore, in order to confirm the inhibitory effect of Tdp43 aggregation according to the concentration of the compounds selected according to the above Experimental Example 3, the selected compounds were treated with Forskolin (treatment concentration 30 μM), a Tdp43 aggregation inducer, at concentrations of 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3, and 10 μM, respectively, to tau-BiFC cells. After 48 hours, the cell images were checked to analyze the Tdp43 aggregation reaction and the degree of cytotoxicity. IC of the compounds 50 And toxicity was analyzed using nonlinear regression analysis using Prism software (GraphPad). The results for representative compounds are shown in Table 4: TDP-43 protein aggregation inhibition effect and cell activity of novel compounds.

[1299] Compound numberTDP-43 protein aggregation inhibition effect (TDP-43 BiFC cells) Compound numberTDP-43 protein aggregation inhibition effect (TDP-43 BiFC cells) Aggregation inhibition effect (%) (@3 uM) IC 50 (nM) Cell viability (%) (@ 3 uM) Aggregation inhibition effect (%) (@ 3 uM) IC 50(nM) Cell viability (%) (@ 3 uM) 178.187.5210087.5379.6102.34100104.2594.7109.2697.498.1810083.0910076.21010083.81110080.91210073.01310063.614100101.4159 7.5105.21610097.81710096.91898.6102.11992.697.02091.714075.42266.5130.72399.210781.12497.95383.12598.25376.72694.824876.127 48.2104.82857.1113.42947.296.23088.014683.13141.42,90182.73295.49959.73393.912455.03482.41059.53562.919267.43791.20.761.338 49.6111.93958.7117.74067.795.64187.871.54264.762.44392.989.04471.185.04581.137173.64681.432073.6Comparative compound 172.979.0Comparative compound 669.560.8

[1300]

[1301] Experimental Example 5: Confirmation of the liver microsomal stability of a novel compound.

[1302] The liver microsome stability of the compounds prepared according to Examples 1 to 46 was confirmed. Specifically, four types of liver microsomes (0.5 mg mL each from human, dog, rat, and mouse) and 0.1 M phosphate buffer solution (pH 7.4) were added, and the compounds were added at a concentration of 1 μM, and pre-incubated at 37°C for 5 minutes. Then, an NADPH generation system solution was added and incubated at 37°C for an additional 30 minutes. Afterwards, an acetonitrile solution containing an internal standard (chloropropamide) was added to terminate the reaction, and after centrifugation for 5 minutes (14,000 rpm, 4°C), the supernatant was injected into an LC-MS / MS system to analyze the substrate drug, thereby evaluating the metabolic stability of eight compounds.

[1303] The amount of substrate remaining through the above reaction was analyzed using a Shimadzu Nexera XR system and TSQ vantage (Thermo). The HPLC column used was Kinetex XB-C18 (2.1 × 100 mm, 2.6 μm, particle size; Phenomenex, USA), and the mobile phases used were (A) distilled water containing 0.1% formic acid and (B) acetonitrile containing 0.1% formic acid. Analyst software (version 1.6.3) and Xcalibur (version 1.6.1) were used for data analysis. The results are presented in Table 5: Stability data of four liver microsomes (human, rat, and mouse) of the novel compounds.

[1304] Compound #Human Microsomes (Residual Rate %)Rat Microsomes (Residual Rate %)Mouse Microsomes (Residual Rate %)191.22.149.3274.355.294.10399.473.080.6493.580.998.5577.877.891.969066.187.3883.976.677.495.017.783.81069.484.887.31181.373.987.81262.770.185.11379.947.784.815>10060>1001679.849.689.51716.759.3>1001 895.872.986.220>10082.995.42385.669.886.62696.395.1992811.750.781.830>10091.983.83199.988.3>1003295.581.792.33394.996.8893488.997.178.23594.374.195.13887.696.086.13983.882.388.44094.287.793.44170.019.065.3Comparative compound 137.01.21.2Comparative compound 684.783.9>100

[1305]

[1306] Experimental Example 6: Inhibitory effect of novel compounds on CYP isoenzyme activity

[1307] The inhibitory effect on CYP isoenzyme activity by compounds prepared according to Examples 1 to 46 was confirmed. Specifically, human liver microsomes (0.25 mg / mL), 0.1 M phosphate buffer solution (pH 7.4), a cocktail of substrate drugs of five drug-metabolizing enzymes (Phenacetin 50 μM, Diclofenac 10 μM, S-mephenytoin 100 μM, Dextromethorphan 5 μM, and Midazolam 2.5 μM), and compounds were added at concentrations of 0 or 10 μM, respectively, and pre-incubated at 37°C for 5 minutes. After adding the NADPH generation system solution, the mixture was further incubated at 37°C for 15 minutes. Afterwards, the reaction was terminated by adding an acetonitrile solution containing an internal standard (Terfenadine), and after centrifugation for 5 minutes (14,000 rpm, 4℃), the supernatant was injected into an LC-MS / MS system to simultaneously analyze metabolites of the substrate drug, thereby evaluating the drug-metabolizing enzyme inhibition ability.

[1308] The metabolites of each CYP isoenzyme marker drug generated through the above reaction were analyzed using a Shimadzu Nexera XR system and TSQ vantage (Thermo). The HPLC column used was a Kinetex C18 (2.1 × 100 mm, 2.6 μm, particle size; Phenomenex, USA), and the mobile phases used were (A) distilled water containing 0.1% formic acid and (B) acetonitrile containing 0.1% formic acid, and the gradient program shown in Table 6 below was applied.

[1309] Time (min)Flow rate (mL / min)%A%B00.310001.00.360404.00.350504.10.310007.00.31000

[1310]

[1311] The generated metabolites were quantified using MRM (multiple reaction monitoring) quantitative mode, and Xcalibur (version 1.6.1) was used for data analysis. In order to show the inhibitory effect of the novel compounds manufactured according to the examples of the present invention on CYP isoenzyme activity, the % activity of CYP isoenzyme with respect to the control group to which no compound was added is shown in Table 6 below. Additionally, in order to confirm the difference in effect according to the substituent, two compounds, Comparative Compounds 1 and 5, which share a similar parent nucleus structure as Compounds 1 to 46 of the present invention, but in which Cy is substituted with aromatic heteroaryl rather than saturated heterocyclyl, were selected as comparative compounds, and their activities were also shown.

[1312] Compound numberCYP1A2(% activity)CYP2C9(% activity)CYP2C19(% activity)CYP2D6(% activity)CYP3A4(% activity)180.282.5>100>10095.5281.795.196.7>10095.7399.9>100>100>100>100486.893.293.4>100945 84.39890.2>10086.1692.3>10098.4>10096.7757.667.895.1>10083.1891.8>100>100>10095968.960.786.9> 10066.11068.476.883.693.874.51183.770.280.398.678.41274.965.184.894.878.913837583.3>10084.81489.78895.5>10096.81580.288.687.9>10087.31699.987>10092.196.71783.273.89089.587.11898.9>100100 9794.62055.64851.857.555.42370.668.569.671.571.72465.172.067.974.274.7258793.585.793.591.52679.153.382.182.181.32795.158.595.492.359.82895.548.590.87887.83088.986.286.288.974.63192.788. 890.893.388.33278.965.177.192.1733375.528.878.697.975.43485.880.880.692.484.33581.584.38294.986.13883.973.68095.786.7398442.978.685.956.54080.246.965.788.675.54171.951.286.786.886.5Comparative compound 680.232.081.291.881.7

[1313]

[1314] Experimental Example 7: Pharmacokinetic Study of a Novel Compound

[1315] The pharmacokinetics of the compounds prepared according to Examples 1 to 46 were studied using an experimental animal model. Specifically, 7-8 week-old SD rats (Coatech, Hana Sangsa Co., Ltd.) weighing approximately 250 to 300 g were purchased and maintained in a small animal simple breeding room set at a temperature of 22±2℃, a relative humidity of 50±5%, a lighting period of 12 hours (lights on at 8:00 AM and lights off at 8:00 PM), and an illumination intensity of 150 to 300 lux. During the entire experimental period, feed and RO water were provided ad libitum, and fasting was performed for 16 hours before oral drug administration. After preparing the drugs under the conditions shown in Table 7 below, the drugs were administered intravenously (IV) or orally (PO). Blood transfusions were collected in 500 μL aliquots into tubes containing heparin sodium and centrifuged at 8,000 rpm for 6 minutes at 2 to 8°C to obtain plasma. Twenty μL of the obtained serum was added to 180 μL of acetonitrile containing an internal standard, vortexed, and centrifuged at 15,000 rpm for 5 minutes at 4°C. The resulting supernatant was analyzed by LC-MS / MS. The HPLC and MS conditions used for the analysis are shown in Tables 8 and 9, respectively.

[1316] Experimental group Administration route Formulation Dose (mg / kg) Dose volume (μL) 1 IV 10% solutol, 10% PEG400 80% DW (1% 1N NaOH) 1250 2 PO 10% solutol, 10% PEG400 80% DW (1% 1N NaOH) 15500

[1317]

[1318] HPLC system: Nexera XR system (Shimadzu, Japan) Column: Kinetex C18 column (2.1 × 100 mm, 2.6 μm, particle size; Phenomenex, USA) Injection volume: 2 μL Mobile phase: (A) 0.1% formic acid aqueous solution (B) 0.1% formic acid acetonitrile solution Sample analysis time: 3.5 min Retention time: 2.2 min

[1319]

[1320] Analytical System TSQ vantage triple quadruple (Thermo, USA) Molecular Weight 312.35 g / mol Ion Source Type & Ionization Mode Turbo Spray Ionization, negative mode - MRM transition (m / z): 263.090 → 185.1 - CE (V): 19, S-lens: 83 Lower Limit of Quantification (LLOQ) - Plasma: 1 ng / mL - Brain: 1 ng / mL Standard Curve Range - Plasma: 1 to 5000 ng / mL - Brain: 1 to 2000 ng / mL

[1321]

[1322] Specific PK variables were calculated using the Phoenix WinNonlin 6.4 version (Pharsight, USA) program and the non-compartmental analysis model, and the results are summarized in Table 11 below.

[1323] Compound Serum PK Parameters (Plasma PK Parameters) Administration Route Dose (mg / kg) AUC all (hours ng / mL)T 1 / 2 (Time)C max (ng / mL)T max(Time) CL (mL / min / kg) F (%) 6 IV 2 1 7 0 1.34 0.58 9 8 6 8.51-19.54 2.0 PO 10 3,569.920.894 8 8 1.4 10.25-16 IV 2 4,090.37 0.514092.93-8.33 113.9 PO 10 2,312.94 1.87 11,145.04 0.67-20 IV 1 1 3 0.10.2--131.5 113.5 PO 15 2,214.21.6 20 46.60.4

[1324]

[1325] Experimental Example 8: Analysis of the cognitive function improvement effect of a novel compound in a tau genetically modified animal model (TauP301L-BiFC Tg mouse).

[1326] Compounds 6, 16, and 20 were orally administered five times a week at a concentration of 10 mg / kg to 8.2-month-old tau transgenic animal models (TauP301L-BiFC tg mice) for a total of 19 weeks (n=10 / group). The vehicle-treated group of TauP301L-BiFC mice and WT mice of similar age were used as controls. To confirm the effects of drug administration on cognitive and motor functions, three behavioral tests (Novel Objective Recognition test, Y-maze test, Balance beam test) were performed, and the results of these tests and each were as follows.

[1327] (1) Novel Objective Recognition test: To evaluate the object recognition memory ability of mice, they were allowed to explore two identical objects (Old-Old) for 10 minutes in a 40cmx40cm box. The next day, one object was replaced with a new object (Old-New) and the mice were allowed to explore for another 10 minutes. The time it took for the exploration behavior toward the existing object and the new object was measured. Mice with normal object recognition ability (Recognition index) took a long time to explore the new object, whereas mice with low object recognition ability could not distinguish the new object. The object recognition index values ​​were as follows. (RI, Vehicle= 0.48, ns, #20= 0.59, p<0.05, #6= 0.66, p<0.0001, #16= 0.58, p<0.05). In TauP301L-BiFC mice, the effects of drugs #20, #6, and #16 on improving object recognition function were confirmed compared to the vehicle group.

[1328] (2) Y-maze test: The Y-maze test was performed to evaluate the spatial working memory ability of mice. Video recordings were used to measure whether the mice spontaneously alternated between the three branches of the Y-maze for a certain period of time. An increase in the spontaneous alternation ratio was considered to indicate improved working memory ability. The spontaneous alternation ratios were as follows. (Alteration ratio (%), vehicle = 60.7%, #20 = 70.8%, p<0.05, #6 = 71.1%, p<0.05, #16 = 76.4%, p<0.001). In TauP301L-BiFC mice, the spatial working ability improvement effect of drugs #20, #6, and #16 was confirmed compared to the vehicle group.

[1329] (3) Balance beam test: The Balance beam test was performed to evaluate the balance and motor coordination ability of the mice. The time taken to pass through a bar that was 50 cm high and 80 cm long and the number of times the feet went out of the bar during the passage were measured. (Average time taken to pass the bar (sec), vehicle = 10.5 s, ##20 = 8.0 s, ns, #6 = 7.9 s, ns, #16 = 6.7 s, p<0.05), (Average number of times the feet went out of the bar (n), vehicle = 1.5, ##20 = 0.7, p<0.01, #6 = 1.1, ns, #16 = 0.8, p<0.05). The effects of drugs #20, #6, and #16 on improving balance and motor ability were confirmed in TauP301L-BiFC mice compared to the vehicle group.

[1330] The experimental process and results described above are shown in Figure 1.

[1331]

[1332] Experimental Example 9: Analysis of the effect of a novel compound on improving tau pathology in a tau genetically modified animal model (TauP301L-BiFC Tg mouse).

[1333] After oral administration of compounds 6, 16, and 20 and completion of behavioral experiments, 13.2-month-old TauP301L-BiFC mice were sacrificed, and [1] brain tissue was extracted and [2] brain lysate was purified.

[1334] [1] First, brain tissue extraction, BiFC fluorescence imaging of brain tissue, and immunofluorescence of tau phosphorylation antibodies were performed as follows. TauP301L-BiFC mice were perfused with 0.9% saline and fixed in phosphate-buffered saline (PBS) containing 4% paraformaldehyde (pH 7.4). The brains were extracted and fixed in 4% paraformaldehyde in PBS at 4°C for 48–72 hours. For cryoprotection, the brains were transferred to PBS containing 30% sucrose and stored at 4°C until they settled. For cryosectioning, the brains were frozen in OCT (Tissue-TEK) and serially sectioned using a brain tissue slicer. Tissue sections of 30 μm thickness were stored in 0.05% sodium azide in PBS at 4°C.

[1335] Brain tissue sections (n=10 per group) were mounted on slides and stained with Sudan Black B solution (70% ethanol containing 0.05% Sudan Black B) for 10 min to reduce autofluorescence, washed three times with PBS containing 0.1% Triton X-100, and then washed with distilled water. For nuclear staining, brain tissue sections were stained with Hoechst (0.5 μg / mL) for 30 min. BiFC fluorescence (λ ex =460-490nm and λ em =500-550 nm) images were acquired. For quantitative analysis, the average BiFC-fluorescence intensity was measured in the somatosensory cortex (layer V) and hippocampal CA1, and the habenular region was used as a control region to reduce background fluorescence differences between brain samples. Regions of interest were marked using ImageJ software (NIH), and fluorescence intensity values ​​were calculated, and data are presented as the mean fluorescence intensity and standard deviation of brain tissue images of 10 mice per group.

[1336] Tau-BiFC fluorescence imaging results of four regions of total brain tissue (hippocampus (CA1), sensory cortex, cerebellum, and brainstem) showed that the levels of Tau-BiFC fluorescence, which are indicated by the formation of initial oligomers and aggregates of tau aggregation, were as follows.

[1337] (1) Hippocampus (CA1), vehicle = 100%, #20= 79%, p<0.05, #6= 80%, p<0.05, #16= 72%, p<0.001.

[1338] (2) Sensory cortex (SCTX), vehicle = 100%, #20= 72%, p<0.05, #6= 73%, p<0.05, #16= 66%, p<0.01.

[1339] (3) Cerebellum (CB), vehicle = 100%, #20= 64%, p<0.05, #6= 68%, ns, #16= 76%, ns

[1340] (4) Brain stem, vehicle = 100%, #20= 60%, p<0.05, #6= 62%, p<0.05, #16= 60%, p<0.05.

[1341] In TauP301L-BiFC mice, the inhibitory effects of compounds 6, 16, and 20 on tau oligomer formation and improvement of tau pathology compared to vehicle were confirmed, and this is shown in Figure 2.

[1342]

[1343] To measure the degree of tau phosphorylation seen in tau pathology as well as tau oligomer formation, immunofluorescence staining using a tau phosphorylation antibody was performed. For brain tissue immunofluorescence image analysis, brain tissue slices (n=10 per group) were stained with AT8 (pS202 / T205) antibody (1:200), and the primary antibody was detected using Alexa Flour 633-labeled anti-mouse secondary antibody (1:500). Fluorescence images (λ) were captured using an Axio Scan.Z1 (ZEISS). ex=620-640nm and λ em =650-700nm) was obtained.

[1344] The AT8 fluorescence imaging results of two brain tissue regions (hippocampus (CA1) and sensory cortex) were as follows.

[1345] (1) Hippocampus (CA1), vehicle = 100%, #20= 72%, p<0.01, #6= 81%, p<0.05, #16= 76%, p<0.01.

[1346] (2) Sensory cortex (SCTX), vehicle = 100%, #20= 54%, p<0.0001, #6= 63%, p<0.001, #16= 57%, p<0.0001.

[1347] In TauP301L-BiFC mice, the improvement of tau pathology through inhibition of tau phosphorylation by compounds 6, 16, and 20 compared to vehicle was confirmed, and this is shown in Figure 3.

[1348]

[1349] [2] To verify the tau pathology inhibitory efficacy of the drug confirmed through brain tissue imaging, the brain lysate was purified and immunoblotted using a tau antibody on the soluble and insoluble fractions. The brain was weighed, and RIPA lysis buffer containing the corresponding dose of protease and phosphatase inhibitors was added to the brain tissue. The tissue was then homogenized, incubated at 4°C for 2 hours, and centrifuged at 13,200 rpm for 20 minutes at 4°C. After centrifugation, the supernatant was collected as the RIPA-soluble fraction and stored at -80°C. To prepare the RIPA-insoluble fraction, the remaining pellet was washed once with RIPA lysis buffer containing 1 M sucrose, then incubated with 2% SDS solution (1 mL per 1 g of tissue) at room temperature for 1 h. The mixture was then centrifuged at 13,200 rpm for 1 min at room temperature, and the supernatant was collected as the RIPA-insoluble fraction and stored at -80°C.

[1350] For tau immunoblot analysis, 20 μg of RIPA-soluble brain tissue samples were separated on a 10% SDS-PAGE gel and transferred to PVDF membranes. RIPA-insoluble samples were diluted 10-fold in 2% SDS solution, and 20 μL of each sample was loaded onto an SDS-PAGE gel. Total and phosphorylated tau levels were detected with anti-tau antibody Tau5 and anti-phosphorylated tau antibodies pS199 and pS396. Band intensities were quantified using ImageJ software (NIH).

[1351] The results of two types of tau phosphorylation immunoblots using RIPA-soluble brain lysates were as follows.

[1352] (1) pS199: vehicle = 100%, #20= 60%, p<0.05, #6= 48%, p<0.01, #16= 24%, p<0.001.

[1353] (2) pS396: vehicle = 100%, #20= 71%, p<0.05, #6= 60%, p<0.01, #16= 40%, p<0.001.

[1354] Immunoblot results using brain lysates from TauP301L-BiFC mice confirmed the improvement of tau pathology through inhibition of tau phosphorylation by compounds 6, 16, and 20 compared to vehicle in the soluble fraction.

[1355] Total tau immunoblot results using RIPA-insoluble brain lysate were as follows.

[1356] Total tau (Tau5): vehicle = 100%, #20= 23%, p<0.0001, #6= 14%, p<0.0001, #16= 20%, p<0.0001.

[1357] Immunoblot results using brain lysates from TauP301L-BiFC mice confirmed the improvement of tau pathology through inhibition of insoluble tau aggregate formation by compounds 6, 16, and 20 compared to vehicle in the insoluble fraction.

[1358] The results of the above experiment are shown in Fig. 4.

[1359]

[1360] Although the embodiments described above have been described with limited drawings, those skilled in the art will appreciate that various technical modifications and variations can be applied based on the above. For example, appropriate results can still be achieved even if the described techniques are performed in a different order than described, and / or components of the described systems, structures, devices, circuits, etc. are combined or combined in a different manner than described, or are replaced or substituted with other components or equivalents.

[1361] Therefore, other implementations, other embodiments, and equivalents to the claims also fall within the scope of the claims described below.

Claims

1. A carbonohydrazonoyl dicyanide compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, n is 0 or 1; R1 is hydrogen, C 1-4 Alkyl, or C 1-4 Any one selected from alkylcarbonyl; R2 is absent or halogen; X is any one of O, S or NR3; R3 is hydrogen, or C 1-4 It is alkyl; Y1, Y2 are C or N; Z is absent or NH; Cy is unsubstituted or halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylcarbonyl, and C 1-4 A 5 to 10 membered saturated heterocyclyl substituted with at least one selected from the group consisting of alkoxycarbonyl.

2. In paragraph 1, R1 is any one selected from hydrogen, methyl or acetyl; R2 is absent or chlorine; X is any one of O, S or NR3; R3 is any one selected from hydrogen, methyl, or ethyl; A carbonohydrazonoyl dicyanide compound or a pharmaceutically acceptable salt thereof, wherein Cy is a 5 to 10 membered saturated heterocyclyl substituted with one or more selected from the group consisting of fluoro, methyl, isopropyl, methoxy, methylcarbonyl, and tert-butoxycarbonyl.

3. In paragraph 1, The carbonohydrazonoyl dicyanide compound represented by the above [chemical formula 1] is a carbonohydrazonoyl dicyanide compound represented by the following [chemical formula 1-1] or [chemical formula 1-2], or a pharmaceutically acceptable salt thereof. [Chemical Formula 1-1] [Chemical Formula 1-2] .

4. In paragraph 1, The above compound is 1. (2-morpholinobenzo[d]thiazol-5-yl)carbonohydrazonoyl dicyanide, 2. (2-morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide, 3. (2-morpholinobenzo[d]oxazol-5-yl)carbonohydrazonoyl dicyanide, 4. (2-morpholinobenzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide 5. (1-methyl-2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide, 6. (1-methyl-2-morpholino-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide, 7. (2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide 8. (2-(4-methylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 9. (2-(4-methoxypiperidin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 10. (2-(2,6-dimethylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 11. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 12. (2-(2-methylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide 13. (2-(3-methylmorpholino)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 14. (2-(4-methylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide) 15. (2-(2-methylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide 16. (2-(3-methylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide 17. (2-(4-methoxypiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide) 18. (2-(2,6-dimethylmorpholino)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide) 19. (5-chloro-2-morpholinobenzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide 20. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide) 21. tert-butyl 4-(6-(2-(dicyanomethylene)hydrazinyl)benzo[d]oxazol-2-yl)piperazine-1-carboxylate 22. tert-butyl 4-(6-(2-(dicyanomethylene)hydrazinyl)benzo[d]thiazol-2-yl)piperazine-1-carboxylate 23. (2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 24. (2-(piperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 25. (2-(piperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide) 26. (2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide) 27. (2-(4-isopropylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 28. (2-(4-isopropylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide) 29. (2R,6S)-tert-butyl 4-(6-(2-(dicyanomethylene)hydrazinyl)benzo[d]thiazol-2-yl)-2,6-dimethylpiperazine-1-carboxylate 30. (2-(4-acetylpiperazin-1-yl)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 31. (2-(4-acetylpiperazin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide) 32. (1-ethyl-2-morpholino-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide, 33. (1-ethyl-2-morpholino-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide, 34. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-6-yl)carbonohydrazonoyl dicyanide) 35. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1-methyl-1H-benzo[d]imidazol-5-yl)carbonohydrazonoyl dicyanide) 36. (2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)benzo[d]thiazol-6-yl)(methyl)carbonohydrazonoyl dicyanide) 37. Acetyl(2-morpholinobenzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide 38. (2-(morpholine-4-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 39. (2-(2,6-dimethylmorpholine-4-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 40. (2-(8-oxa-3-azabicyclo[3.2.1]octane-3-carboxamido)benzo[d]thiazol-6-yl)carbonohydrazonoyl dicyanide) 41. (R)-(2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide 42. (S)-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide 43. (S)-(2-(3-fluoropyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide 44. (2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide) 45. (2-(pyrrolidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide 46. ​​(2-(piperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide 47.(R)-N-(2-(3-fluoropiperidin-1-yl)benzo[d]oxazol-6-yl)carbonohydrazonoyl dicyanide 48.N-(2-(morpholine-4-carbonyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)carbonohydrazonoyl dicyanide, or 49. A carbonohydrazonoyl dicyanide compound, which is N-(2-morpholinooxazolo[4,5-b]pyridin-6-yl)carbonohydrazonoyl dicyanide, or a pharmaceutically acceptable salt thereof.

5. A first step of forming an imine bond by reacting a compound represented by the following chemical formula 2 containing a reactive amine group at one terminal with sodium nitrite and malononitrile in the presence of an acid; and Optionally, a method for producing a compound represented by the chemical formula 1, further comprising a second step of introducing an R1 substituent into the product obtained from the first step, when R1 is a substituent other than hydrogen: [Chemical Formula 1] [Chemical Formula 2] In the above chemical formulas 1 and 2, n is 0 or 1; R1 is hydrogen, C 1-4 Alkyl, or C 1-4 Any one selected from alkylcarbonyl; R2 is absent or halogen; X is any one of O, S or NR3; R3 is hydrogen, or C 1-4 It is alkyl; Y1, Y2 are C or N; Z is absent or NH; Cy is unsubstituted or halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylcarbonyl, and C 1-4 A 5 to 10 membered saturated heterocyclyl substituted with at least one selected from the group consisting of alkoxycarbonyl.

6. In paragraph 5, The second step above is, i) R1 is C 1-6 In the case of alkyl, it is carried out by reacting with a halogenated alkane in the presence of potassium tert-butoxide or sodium hydride in an organic solvent, ii) R1 is C 1-4 A manufacturing method, which is performed by reacting with a halogenated alkylcarbonyl in the presence of a base and triethylamine, in the case of an alkylcarbonyl.

7. In paragraph 5, The compound represented by the above chemical formula 2 is i) When n is 0 and X is S or NR3, irradiate with microwaves to react Cy-H with a compound represented by the chemical formula 3-a, ii) When n is 0 and X is O, react Cy-H with a compound represented by chemical formula 3-b in the presence of triethylamine, iii) When n is 1, react Cy-H with a compound represented by chemical formula 3-c, R N In this case, the reaction product is further reduced, iv) A manufacturing method obtained by reacting a compound represented by 3-d, where Y2 is N, n is 1, and Z is NH: [Chemical Formula 3-a] [Chemical Formula 3-b] [Chemical Formula 3-c] [Chemical Formula 3-d] In the above chemical formulas 3-a to 3-d, X1 is halogen, R N is nitro or amino; R5 is any one selected from t-butoxy carbonyl, hydrogen, or benzyloxy carbonyl; Cy is unsubstituted or halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylcarbonyl, and C 1-4 A 5 to 10 membered saturated heterocyclyl substituted with at least one selected from the group consisting of alkoxycarbonyl.

8. In paragraph 7, In the compound represented by the above chemical formula 3-a, X is NR3 and R3 is C 1-4 A method for preparing an alkyl compound, wherein the compound in which R3 is hydrogen is prepared by the step i) above and reacting the compound with halogenated R3 in the presence of DBU (1,8-Diazabicyclo[5.4.0]undec-7-ene).

9. In paragraph 7, The compound represented by the above chemical formula 3-b is prepared by reacting the compound represented by the following chemical formula 4 with potassium ethyl xanthogenate, the manufacturing method: [Chemical Formula 4] .

10. In paragraph 7, The compound represented by the above chemical formula 3-c is prepared by reacting the compound represented by the following chemical formula 5 with phenylchloroformate in the presence of triethylamine, the manufacturing method: [Chemical Formula 5] .

11. A pharmaceutical composition for preventing or treating a disease caused by aggregation or hyperphosphorylation of tau protein or Tdp-43 protein, comprising the compound of paragraph 1 as an active ingredient.

12. A pharmaceutical composition according to claim 11, wherein the disease caused by aggregation or hyperphosphorylation of the tau protein is selected from the group consisting of Alzheimer's disease, Parkinson's disease, vascular dementia, acute stroke, trauma, cerebrovascular disease, brain cord trauma, spinal cord trauma, peripheral neuropathy, retinopathy, glaucoma, and tauopathies.

13. In paragraph 12, The above tauopathies include chronic traumatic encephalopathy (CTE), primary age-related tauopathy, progressive supranuclear palsy, corticobasal degeneration, Pick's disease, argyrophilic grain disease (AGD), frontotemporal dementia (FTD), Parkinsonism linked to chromosome 17, Lytico-Bodig disease (Parkinson-dementia complex of Guam), ganglioglioma, gangliocytoma, meningioangiomatosis, postencephalitic parkinsonism, and subacute sclerosing encephalopathy. A pharmaceutical composition selected from the group consisting of subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, lipofuscinosis, posttraumeric stress disorder, and traumatic brain injury.

14. In paragraph 11, Diseases caused by aggregation or hyperphosphorylation of the TDP-43 protein include amyotrophic lateral sclerosis (ALS), familial amyotrophic lateral sclerosis (Familial ALS-SOD1) due to genetic mutation, sporadic amyotrophic lateral sclerosis, superoxide dismutase 1 (SOD1), frontotemporal lobar degeneration (FTLD), amyotrophic lateral sclerosis with frontotemporal dementia (ALS-FTLD), multiple system proteinopathy (MSP), limbic-predominant age-related TDP-43 encephalopathy (LATE) / CART (cerebral age-related TDP-43 with sclerosis), A pharmaceutical composition selected from the group consisting of Perry disease, facial onset sensory and motor neuronopathy (FOSMN), and sporadic inclusion body myositis (sIBM).

15. A pharmaceutical composition according to claim 11, characterized in that the composition inhibits aggregation of tau protein or Tdp-43 protein.

16. A pharmaceutical composition according to claim 11, characterized in that the composition inhibits hyperphosphorylation of tau protein or Tdp-43 protein.

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