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Patent Searching and Data


Title:
COMPOUNDS, COMPOSITIONS AND METHODS OF USE
Document Type and Number:
WIPO Patent Application WO/2020/117877
Kind Code:
A1
Abstract:
Herein, compounds, compositions and methods for modulating inclusion formation and stress granules in cells related to the onset of neurodegenerative diseases, musculoskeletal diseases, cancer, ophthalmological diseases, and viral infections are described.

Inventors:
VACCA JOSEPH (US)
WAGER TRAVIS (US)
Application Number:
PCT/US2019/064356
Publication Date:
June 11, 2020
Filing Date:
December 04, 2019
Export Citation:
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Assignee:
AQUINNAH PHARMACEUTICALS INC (US)
International Classes:
A61K31/425; A61K31/427
Foreign References:
US20070060566A12007-03-15
US20100317643A12010-12-16
US20170135992A12017-05-18
US20050282862A12005-12-22
US20030212068A12003-11-13
Other References:
DATABASE PUBCHEM 29 February 2008 (2008-02-29), "2-Chloro-5-(1,1-dioxido-1,2-thiazinan-2-yi)-N-(3,4,5-trimethoxyphenyl)benzamide", retrieved from ncbi
Attorney, Agent or Firm:
DENHART, Derek et al. (US)
Download PDF:
Claims:
We claim:

1. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and compound of Formula (I):

Formula (I)

or a pharmaceutically acceptable salt thereof, wherein:

Ring A is aryl or heteroaryl, optionally substituted with 1-5 R9;

R1 is H, Ci-C6 alkyl or CF -CF haloalkyl;

R2 is Ci-Ce alkyl, Ci-Ce alkenyl, Ci-Ce alkynyl, Ci-Ce heteroalkyl, Ci-Ce haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -ORB, -C(0)RD, -C(0)ORB, -NRARC, -NRAC(0)RD, -S(0)xRE, - OS(0)xRE, -C(0)NRAS(0)XRe, -NRAS(0)XRe, or -S(0)xNRA;

each of R3, R4, and R5 is independently H, C i -CF alkyl, CF-CF alkenyl, CF-CF alkynyl, Ci- Ce heteroalkyl, C i -O, haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -ORB, -C(0)RD, - C(0)ORB, -NRARc, -NRAC(0)RD, -S(0)xRE, -OS(0)xRE, -C(0)NRAS(0)xRE, -NRAS(0)xRE, or -S(0)xNRA;

R6 is Ci-Ce alkyl optionally substituted with 1-4 R8;

R7 is Ci-Ce alkyl or -N(R10)-CI-C6 alkyl, each of which is optionally substituted with 1-4 R8;

or R6 and R7, taken together with the atoms to which they are attached, form a 5-7 membered heterocyclic ring, optionally substituted with 1-5 R8, wherein said heterocyclic ring either includes no heteroatoms other than the N and S to which R6 and R7 are attached, or includes one additional N ring atom substituted with R10;

each R8 is independently halo, CF-CF alkyl, -ORB, -C(0)ORB, or Ci-Ce haloalkyl;

each R9 is independently C i -O, alkyl, CF-CF, alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, heterocyclyl, aryl, heteroaryl, -ORB, - C(0)RD, -C(0)ORB, -NRARC -NRAC(0)RD, -S(0)xRE, -OS(0)xRE, -C(0)NRAS(0)xRE, - NRAS(0)XRe, or -S(0)xNRA; wherein each cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted by 1-5 R11;

or two R9groups bound to adjacent ring atoms of Ring A, taken together with the atoms to which they are attached, form a carbocyclic or heterocyclic ring with Ring A (e.g., a 5-7 membered ring, e.g., a cyclopentyl ring fused to Ring A or a piperidinyl ring fused to Ring A). each R10 is independently H, C1-C6 alkyl or C1-C6 haloalkyl;

each R11 is independently C1-C6 alkyl, Ci-Ce alkenyl, CYO, alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, oxo, cycloalkyl, -ORB, -C(0)RD, -C(0)0RB, -NRARC, -NRAC(0)RD, -S(0)xRE, -0S(0)xRE, -C(0)NRAS(0)xRE, -NRAS(0)xRE, or -S(0)xNRA

each RA, RB, Rc, RD, or RE is independently H, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1-4 R8;

or RA and Rc, together with the atoms to which each is attached, form a heterocyclyl ring optionally substituted with 1-4 R8; and

x is 0, 1, or 2.

2. The pharmaceutical composition of claim 1, wherein

wherein m is 0, 1, 2, 3, or 4.

3. The pharmaceutical composition of claim 1, wherein

wherein m is 0, 1, 2, 3, or 4.

4. The pharmaceutical composition of claim 1, wherein

R6 the u O moiety is

wherein m is 0 or 1.

5. The pharmaceutical composition of any one of claims 2-4, wherein m is 0.

6. The pharmaceutical composition of any one of the preceding claims, wherein Ring A is aryl (e.g., phenyl).

7. The pharmaceutical composition of any one of the preceding claims, wherein Ring A is phenyl.

8. The pharmaceutical composition of any one of the preceding claims, wherein Ring A is

9. The pharmaceutical composition of any one of the preceding claims, wherein at least one R9 group is Ci-C6 alkyl or halo (e.g., chloro).

10. The pharmaceutical composition of any one of the preceding claims, wherein at least one R9 group is methyl or chloro.

11. The pharmaceutical composition of claim 8, wherein one R9 is chloro or methyl and the other R9 is G-G alkyl, G-G alkenyl, G-G alkynyl, C i -G alkoxy, G-G haloalkyl, halo, cyano, heterocyclyl, phenyl, or heteroaryl; wherein each heterocyclyl, phenyl, or heteroaryl is optionally substituted by 1-5 R11.

12. The pharmaceutical composition of claim 8, wherein one R9 is chloro and the other R9 is phenyl, monocyclic 4-7 membered heterocyclyl (wherein 1-2 of the ring atoms are heteroatoms selected from the group consisting of N, O, and S), or monocyclic 5-6 membered heteroaryl (wherein 1-2 of the ring atoms are heteroatoms selected from the group consisting of N, O, and S); wherein the phenyl, heterocyclyl, or heteroaryl is optionally substituted with 1-3 R11.

13. The pharmaceutical composition of claim 8, wherein one R9 group is chloro and the other R9 group is selected from the group consisting of

wherein n is 0, 1, 2, 3, 4, or 5.

14. The pharmaceutical composition of claim 13, wherein n is 0.

15. The pharmaceutical composition of any one of claims 1-7, wherein Ring A is

16. The pharmaceutical composition of claim 15, wherein R9 is halo (e.g., chloro).

17. The pharmaceutical composition of claim 15, wherein R9 is G-G alkyl, G-G alkenyl, G-G alkynyl, C i -G heteroalkyl, Ci-G haloalkyl, halo, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, or -ORB; wherein each cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted by 1-5 R11.

18. The pharmaceutical composition of claim 15, wherein R9 is G-G alkyl, G-G alkenyl, G-G alkynyl, G-G heteroalkyl, phenyl, G-G haloalkyl, halo, cyano, or G-G alkoxy.

19. The pharmaceutical composition of any one of claims 1-5, wherein Ring A is heteroaryl.

20. The pharmaceutical composition of claim 19, wherein Ring A is monocyclic 5-6 membered heteroaryl wherein 1-2 of the ring atoms are heteroatoms selected from O, N, and S.

21. The pharmaceutical composition of claim 19, wherein Ring A is monocyclic 5-6 membered heteroaryl wherein 1-2 of the ring atoms are nitrogen heteroatoms.

22. The pharmaceutical composition of claim 19, wherein Ring A is seleted from the group consisting of

23. The pharmaceutical composition of claim 22, wherein p is 0 or 1.

24. The pharmaceutical composition of any one of claims 19-23, wherein each R9 is independently halo, G-G alkyl, G-G alkenyl, G-G alkynyl, G-G heteroalkyl, Ci-G haloalkyl, halo, cyano, or G-G alkoxy.

25. The pharmaceutical composition of claim 19, wherein Ring A is seleted from the group consisting of

26. The pharmaceutical composition of any one of claims 19-25, wherein R1 is H.

27. The pharmaceutical composition of any one of claims 19-26, wherein R2 is halo, G-G alkyl or G-G haloalkyl.

28. The pharmaceutical composition of any one of the preceding claims, wherein R2 is halo.

29. The pharmaceutical composition of any one of the preceding claims, wherein R2 is chloro.

30. The pharmaceutical composition of any one of the preceding claims, wherein R3 is H or halo (e.g., chloro).

31. The pharmaceutical composition of any one of the preceding claims, wherein R4 and R5 are both H.

32. The pharmaceutical composition of any one of the preceding claims, wherein R4 and R5 are each H, R3 is either H or chloro, and R2 is chloro.

33. The pharmaceutical composition of any one of the preceding claims, wherein the compound of Formula (I) is a compound of Formula (I-a):

Formula (I-a)

or a pharmaceutically acceptable salt thereof, wherein Ring A, R^R11, n, x, p, and subvariables thereof are as described in any of the preceding claims, and

wherein W is -NH-, -CH2CH2-, or -CH2NH-, wherein the NH of -CH2NH- is linked to the sulfur atom of Formula (I-a).

34. The pharmaceutical composition of any one of the preceding claims, wherein the compound of Formula (I) is a compound of Formula (I-b):

Formula (I-b)

or a pharmaceutically acceptable salt thereof, wherein R '-R7, R3,R9-Rn, n, x, p, and subvariables thereof are as described in any of the preceding claims, and wherein W is -CH2-, - CH2CH2-, or -CH2NH-, wherein the NFI of -CH2NH- is linked to the sulfur atom of Formula (I- b).

35. The pharmaceutical composition of any one of the preceding claims, wherein the compound of Formula (I) is a compound of Formula (I-c):

Formula (I-c)

or a pharmaceutically acceptable salt thereof, wherein R2, R3, R9-Rn, x, p and subvariables thereof are as described in any of the preceding claims.

36. The pharmaceutical composition of any one of the preceding claims, wherein the compound of Formula (I) is a compound of Formula (I-d):

Formula (I-d)

or a pharmaceutically acceptable salt thereof, wherein wherein R2, R3, R9-Rn, x, p, and subvariables thereof are as described in any of the preceding claims.

37. The pharmaceutical composition of any one of the preceding claims, wherein the compound of Formula (I) is a compound of Formula (I-e):

Formula (I-e)

or a pharmaceutically acceptable salt thereof, wherein R3, R9-Rn, x, p, and subvariables thereof are as described in any of the preceding claims. 38. The pharmaceutical composition of any one of the preceding claims, wherein the compound of Formula (I) is a compound of Formula (I-f):

Formula (I-f)

or a pharmaceutically acceptable salt thereof, wherein R9-Rn, x, p, and subvariables thereof are as described in any of the preceding claims, and wherein R3 is H or chloro.

39. The pharmaceutical composition of any one of the preceding claims, wherein the compound of Formula (I) is selected from a compound described in FIGS. 1A-1TT.

40. The pharmaceutical composition of any one of the preceding claims, wherein the compound of Formula (I) is selected from the group consisting of:

41. A method for modulating stress granules, wherein the method comprises administering to a subject in need thereof a pharmaceutical composition of any one of claims 1-40.

42. The method of claim 41, wherein the stress granule comprises tar DNA binding protein- 43 (TDP-43), T-cell intracellular antigen 1 (TIA-1), TIA1 cytotoxic granule-associated RNA binding protein-like 1 (TIAR), GTPase activating protein binding protein 1 (G3BP-1), GTPase activating protein binding protein 2 (G3BP-2), tris tetraprolin (TTP), fused in sarcoma (FUS), or fragile X mental retardation protein (FMRP).

43. A method for modulating TDP-43 inclusion formation, wherein the method comprises administering to a subject in need thereof a pharmaceutical composition of any one of claims 1- 40.

44. The method of any one of claims 41-43, wherein the subject has a neurodegenerative disease or disorder, a musculoskeletal disease or disorder, a cancer, an ophthalmological disease or disorder, and/or a viral infection.

45. The method of claim 44, wherein the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, frontotemporal dementia (FTD), FTLD-U, FTD caused by mutations in the progranulin protein or tau protein (e.g., progranulin-deficient FTLD), frontotemporal dementia with inclusion body myopathy (IBMPFD), frontotemporal dementia with motor neuron disease, amyotrophic lateral sclerosis (ALS), Huntington’s disease (HD), Huntington’s chorea, prion diseases (e.g., Creutzf eld- Jacob disease, bovine spongiform encephalopathy, Kuru, or scrapie), Lewy Body disease, diffuse Lewy body disease (DLBD), polyglutamine (polyQ)-repeat diseases, trinucleotide repeat diseases, cerebral degenerative diseases, presenile dementia, senile dementia, Parkinsonism linked to chromosome 17 (FTDP- 17), progressive supranuclear palsy (PSP), progressive bulbar palsy (PBP), psuedobulbar palsy, spinal and bulbar muscular atrophy (SBMA), primary lateral sclerosis, Pick's disease, primary progressive aphasia, corticobasal dementia, HIV-associated dementia, Parkinson's disease, Parkinson's disease with dementia, dementia with Lewy bodies, Down's syndrome, multiple system atrophy, spinal muscular atrophy (SMA, e.g., SMA Type I (e.g., Werdnig-Hoffmann disease) SMA Type II, SMA Type III (e.g., Kugelberg-Welander disease), or congenital SMA with arthrogryposis), progressive spinobulbar muscular atrophy (e.g., Kennedy disease), post polio syndrome (PPS), spinocerebellar ataxia, pantothenate kinase-associated neurodegeneration (PANK), spinal degenerative disease/motor neuron degenerative diseases, upper motor neuron disorder, lower motor neuron disorder, age-related disorders and dementias, Hallervorden-Spatz syndrome, cerebral infarction, cerebral trauma, chronic traumatic encephalopathy, transient ischemic attack, Lytigo-bodig (amyotrophic lateral sclerosis-parkinsonism dementia), Guam- Parkinsonism dementia, hippocampal sclerosis, corticobasal degeneration, Alexander disease, Apler’s disease, Krabbe’s disease, neuroborreliosis, neurosyphilis, Sandhoff disease, Tay-Sachs disease, Schilder’s disease, Batten disease, Cockayne syndrome, Kearns-Sayre syndrome, Gerstmann-Straussler-Scheinker syndrome and other transmissible spongiform

encephalopathies, hereditary spastic paraparesis, Leigh’s syndrome, demyelinating diseases, neuronal ceroid lipofuscinoses, epilepsy, tremors, depression, mania, anxiety and anxiety disorders, sleep disorders (e.g., narcolepsy, fatal familial insomnia), acute brain injuries (e.g., stroke, head injury), autism, or any combination thereof.

46. The method of claim 44, wherein the musculoskeletal disease is selected from the group consisting of muscular dystrophy, facioscapulohumeral muscular dystrophy (e.g., FSHD1 or FSHD2), Freidrich’s ataxia, progressive muscular atrophy (PMA), mitochondrial

encephalomyopathy (MELAS), multiple sclerosis, inclusion body myopathy, inclusion body myositis (e.g., sporadic inclusion body myositis), post-polio muscular atrophy (PPMA), motor neuron disease, myotonia, myotonic dystrophy, sacropenia, multifocal motor neuropathy, inflammatory myopathies, and paralysis.

47. The method of claim 44, wherein the cancer is selected from the group consisting of breast cancer, a melanoma, adrenal gland cancer, biliary tract cancer, bladder cancer, brain or central nervous system cancer, bronchus cancer, blastoma, carcinoma, a chondrosarcoma, cancer of the oral cavity or pharynx, cervical cancer, colon cancer, colorectal cancer, esophageal cancer, gastrointestinal cancer, glioblastoma, hepatic carcinoma, hepatoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, non-small cell lung cancer, ophthalmological cancer, osteosarcoma, ovarian cancer, pancreas cancer, peripheral nervous system cancer, prostate cancer, sarcoma, salivary gland cancer, small bowel or appendix cancer, small-cell lung cancer, squamous cell cancer, stomach cancer, testis cancer, thyroid cancer, urinary bladder cancer, uterine or endometrial cancer, vulval cancer, or any combination thereof.

48. The method of claim 47, wherein the lymphoma is selected from a B-cell lymphoma or a T-cell lymphoma.

49. The method of claim 48, wherein the B-cell or T-cell lymphoma is selected from the group consisting of diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma, intravascular large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma, mantle cell lymphoma, marginal zone B-cell lymphomas, extranodal marginal B-cell lymphomas, mucosa-associated lymphoid tissue (MALT) lymphomas, modal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, Waldenstrom’s macroglobulinemia, hairy cell leukemia, primary central nervous system (CNS) lymphoma, precursor T-lymphoblastic lymphoma/leukemia, peripheral T-cell lymphoma, cutaneous T-cell lymphoma, smoldering adult T-cell lymphoma, chronic adult T-cell lymphoma, acute adult T-cell lymphoma, lymphomatous adult T-cell lymphoma, angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma nasal type (ENKL), enteropathy-associated intestinal T-cell lymphoma (EATL), and anaplastic large cell lymphoma (ALCL).

50. The method of claim 44, wherein the ophthalmological disease is selected from the group consisting of macular degeneration, age-related macular degeneration, diabetes retinopathy, histoplasmosis, macular hole, macular pucker, Bietti’s crystalline dystrophy, retinal detachment, retinal thinning, retinoblastoma, retinopathy of prematurity, Usher’s syndrome, vitreous detachment, Refsum disease, retinitis pigmentosa, onchocerciasis, choroideremia, Leber congenital amaurosis, retinoschisis, juvenile retinoschisis, Stargardt disease, ophthalmoplegia, or any combination thereof.

51. The method of claim 44, wherein the viral infection is caused by a virus selected from the group consisting of West Nile virus, respiratory syncytial virus (RSV), herpes simplex virus 1, herpes simplex virus 2, Epstein-Barr virus (EBV), hepatitis virus A, hepatitis virus B, hepatitis virus C, influenza viruses, chicken pox, avian flu viruses, smallpox, polio viruses, HIV-1, HIV- 2, Ebola virus, and any combination thereof.

52. The method of any one of claims 44-51 , wherein the subject is a mammal.

53. The method of claim 52, wherein the subject is human.

54. The method of any one of claims 44-53, further comprising the step of diagnosing the subject with the neurodegenerative disease or disorder, musculoskeletal disease or disorder, cancer, ophthalmological disease or disorder, or viral infection prior to onset of said

administration.

55. The method of any one of claims 44-54, wherein pathology of said neurodegenerative disease or disorder, said musculoskeletal disease or disorder, said cancer, said ophthalmological disease or disorder, or said viral infection comprises stress granules.

56. The method of any one of claims 44-55, wherein pathology of said neurodegenerative disease, said musculoskeletal disease or disorder, said cancer, said ophthalmological disease or disorder, or said viral infection comprises TDP-43 inclusions.

57. A compound of Formula (II):

Formula (II)

or a pharmaceutically acceptable salt thereof, wherein:

W is -CH2-, -CH2CH2-, -N(R10)-, or -CH2N(R10)-, wherein the N of -CH2N(R10)- is linked to the sulfur atom of Formula (II);

R1 is H, C1-C6 alkyl or C1-C6 haloalkyl;

R2 is C1-C6 alkyl, G-G alkenyl, G-G alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -ORB, -C(0)RD, -C(0)0RB, -NRARC, -NRAC(0)RD, -S(0)xRE, - 0S(0)xRE, -C(0)NRAS(0)xRE, -NRAS(0)xRE, or -S(0)xNRA;

each of R3, R4, and R5 is independently H, C1-C6 alkyl, CVG, alkenyl, G-G alkynyl, G- G heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -ORB, -C(0)RD, - C(0)0RB, -NRARc, -NRAC(0)RD, -S(0)xRE, -0S(0)xRE, -C(0)NRAS(0)xRE, -NRAS(0)xRE, or -S(0)xNRA; or

each R8 is independently halo, C1-C6 alkyl, -ORB, -C(0)0RB, or C1-C6 haloalkyl;

each R9 is independently C1-C6 alkyl, G-G alkenyl, G-G alkynyl, G-G heteroalkyl, G-G haloalkyl, halo, cyano, nitro, azido, cycloalkyl, heterocyclyl, aryl, heteroaryl, -ORB, - C(0)RD, -C(0)0RB, -NRARC, -NRAC(0)RD, -S(0)xRE, -0S(0)xRE, -C(0)NRAS(0)xRE, - NRAS(0)xRE, or -S(0)xNRA; wherein each cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted by 1-5 R11;

each R10 is independently H, G-G alkyl or G-G haloalkyl;

each R11 is independently G-G alkyl, G-G alkenyl, G-G alkynyl, G-G heteroalkyl, G-G haloalkyl, halo, cyano, nitro, azido, oxo, cycloalkyl, -ORB, -C(0)RD, -C(0)0RB, -NRARC -NRAC(0)RD, -S(0)xRE, -0S(0)xRE, -C(0)NRAS(0)xRE, -NRAS(0)xRE, or -S(0)xNRA; each R12 is independently C1-C6 alkyl, CVG alkenyl, Ci-Ce alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -ORB, -C(0)RD, -C(0)0RB, -NRARC - NRAC(0)RD, -S(0)xRE, -0S(0)xRE, -C(0)NRAS(0)xRE, -NRAS(0)xRE, or -S(0)xNRA;

each RA, RB, Rc, RD, or RE is independently H, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1-4 R8;

or RA and Rc, together with the atoms to which each is attached, form a heterocyclyl ring optionally substituted with 1-4 R8;

x is 0, 1, or 2;

m is 0, 1, 2, 3, or 4;

q is 0, 1, 2, or 3: and

wherein the compound is not

58. The compound of claim 1 , wherein

W is -CH2- or -CH2CH2-; and

m is 0, 1, 2, 3, or 4.

59. The compound of claim 1 , wherein

W is -CH2-; and

m is 0.

60. The compound of any one of claims 57-58, wherein m is 0.

61. The compound of any one of claims 57-60, wherein each R12 is independently C1-C6 alkyl, CVG alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, halo, cyano, or cycloalkyl,

62. The compound of any one of claims 57-60, wherein q is 0.

63. The compound of claim 62, wherein at least one R9 group is halo or C1-C6 alkyl.

64. The compound of claim 63, wherein at least one R9 group is chloro or methyl.

65. The compound of claim 63, wherein at least one R9 group is halo (e.g., chloro).

66. The compound of claim 64, wherein one R9 is chloro or methyl and the other R9 is Ci-Ce alkyl, G-G alkenyl, G-G alkynyl, G-G alkoxy, C i -G haloalkyl, halo, cyano, heterocyclyl, phenyl, or heteroaryl; wherein each heterocyclyl, phenyl, or heteroaryl is optionally substituted by 1-5 R11.

67. The compound of claim 64, wherein one R9 is chloro or methyl and the other R9 is phenyl, monocyclic 4-7 membered heterocyclyl (wherein 1-2 of the ring atoms are heteroatoms selected from the group consisting of N, O, and S), or monocyclic 5-6 membered heteroaryl (wherein 1-2 of the ring atoms are heteroatoms selected from the group consisting of N, O, and S); wherein the phenyl, heterocyclyl, or heteroaryl is optionally substituted with 1-3 R11.

68. The compound of claim 67, wherein one R9 group is chloro or methyl and the other R9 group is selected from the group consisting of

wherein n is 0, 1, 2, 3, 4, or 5.

69. The compound of claim 68, wherein n is 0.

70. The compound of any one of claims 57-69, wherein R1 is H.

71. The compound of any one of claims 57-70, wherein R2 is halo, Ci-Ce alkyl or Ci-Ce haloalkyl.

72. The compound of any one of claims 57-70, wherein R2 is halo.

73. The compound of any one of claims 57-70, wherein R2 is chloro.

74. The compound of any one of claims 57-73, wherein R3 is H or halo.

75. The compound of any one of claims 57-73, wherein R3 is H or chloro.

76. The compound of any one of claims 57-75, wherein R4 and R5 are both H.

77. The compound of any one of claims 57-76, wherein R4 and R5 are each H, R3 is H or chloro, and R2 is chloro.

78. The compoundof any one of claims 57-77, wherein the compound of Formula (II) is a compound of Formula (Il-a):

Formula (II- a)

or a pharmaceutically acceptable salt thereof, wherein wherein R2, R3, R9-Rn, x, and subvariables thereof are as described in any of the preceding claims.

79. The compound of any one of claims 57-78, wherein the compound of Formula (II) is a compound of Formula (Il-b):

Formula (Il-b)

or a pharmaceutically acceptable salt thereof, wherein R3, R9-Rn, x, and subvariables thereof are as described in any of the preceding claims.

80. The compound of any one of claims 57-79, wherein the compound of Formula (II) is a compound of Formula (II-c):

Formula (II-c)

or a pharmaceutically acceptable salt thereof, wherein R3, R9-Rn, x, and subvariables thereof are as described in any of the preceding claims.

81. The compound of any one of claims 57-80, wherein the compound of Formula (II) is selected from a compound described in FIGS. 1A-1TT.

82. The compound of any one of claims 57-81, wherein the compound of Formula (II) is selected from the group consisting of:

83. The compound of any one of claims 57-81, wherein the compound of Formula (II) is selected from the group consisting of:

84. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of any one of claims 57-83.

85. A method for modulating stress granules, wherein the method comprises administering to a subject in need thereof a compound of any one of claims 57-83. 86. The method of claim 85, wherein the stress granule comprises tar DNA binding protein-

43 (TDP-43), T-cell intracellular antigen 1 (TIA-1), TIA1 cytotoxic granule-associated RNA binding protein-like 1 (TIAR), GTPase activating protein binding protein 1 (G3BP-1), GTPase activating protein binding protein 2 (G3BP-2), tris tetraprolin (TTP), fused in sarcoma (FUS), or fragile X mental retardation protein (FMRP).

87. A method for modulating TDP-43 inclusion formation, wherein the method comprises administering to a subject in need thereof a compound of any one of claims 57-83.

88. The method of any one of claims 85-87, wherein the subject has a neurodegenerative disease or disorder, a musculoskeletal disease or disorder, a cancer, an ophthalmological disease or disorder, and/or a viral infection.

89. The method of claim 88, wherein the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, frontotemporal dementia (FTD), FTLD-U, FTD caused by mutations in the progranulin protein or tau protein (e.g., progranulin-deficient FTLD), frontotemporal dementia with inclusion body myopathy (IBMPFD), frontotemporal dementia with motor neuron disease, amyotrophic lateral sclerosis (ALS), Huntington’s disease (HD), Huntington’s chorea, prion diseases (e.g., Creutzf eld- Jacob disease, bovine spongiform encephalopathy, Kuru, or scrapie), Lewy Body disease, diffuse Lewy body disease (DLBD), polyglutamine (polyQ)-repeat diseases, trinucleotide repeat diseases, cerebral degenerative diseases, presenile dementia, senile dementia, Parkinsonism linked to chromosome 17 (FTDP- 17), progressive supranuclear palsy (PSP), progressive bulbar palsy (PBP), psuedobulbar palsy, spinal and bulbar muscular atrophy (SBMA), primary lateral sclerosis, Pick's disease, primary progressive aphasia, corticobasal dementia, HIV-associated dementia, Parkinson's disease, Parkinson's disease with dementia, dementia with Lewy bodies, Down's syndrome, multiple system atrophy, spinal muscular atrophy (SMA, e.g., SMA Type I (e.g., Werdnig-Hoffmann disease) SMA Type II, SMA Type III (e.g., Kugelberg-Welander disease), or congenital SMA with arthrogryposis), progressive spinobulbar muscular atrophy (e.g., Kennedy disease), post polio syndrome (PPS), spinocerebellar ataxia, pantothenate kinase-associated neurodegeneration (PANK), spinal degenerative disease/motor neuron degenerative diseases, upper motor neuron disorder, lower motor neuron disorder, age-related disorders and dementias, Hallervorden-Spatz syndrome, cerebral infarction, cerebral trauma, chronic traumatic encephalopathy, transient ischemic attack, Lytigo-bodig (amyotrophic lateral sclerosis-parkinsonism dementia), Guam- Parkinsonism dementia, hippocampal sclerosis, corticobasal degeneration, Alexander disease, Apler’s disease, Krabbe’s disease, neuroborreliosis, neurosyphilis, Sandhoff disease, Tay-Sachs disease, Schilder’s disease, Batten disease, Cockayne syndrome, Kearns-Sayre syndrome, Gerstmann-Straussler-Scheinker syndrome and other transmissible spongiform

encephalopathies, hereditary spastic paraparesis, Leigh’s syndrome, demyelinating diseases, neuronal ceroid lipofuscinoses, epilepsy, tremors, depression, mania, anxiety and anxiety disorders, sleep disorders (e.g., narcolepsy, fatal familial insomnia), acute brain injuries (e.g., stroke, head injury), autism, or any combination thereof.

90. The method of claim 88, wherein the musculoskeletal disease is selected from the group consisting of muscular dystrophy, facioscapulohumeral muscular dystrophy (e.g., FSHD1 or FSHD2), Freidrich’s ataxia, progressive muscular atrophy (PMA), mitochondrial

encephalomyopathy (MELAS), multiple sclerosis, inclusion body myopathy, inclusion body myositis (e.g., sporadic inclusion body myositis), post-polio muscular atrophy (PPMA), motor neuron disease, myotonia, myotonic dystrophy, sacropenia, multifocal motor neuropathy, inflammatory myopathies, and paralysis.

91. The method of claim 88, wherein the cancer is selected from the group consisting of breast cancer, a melanoma, adrenal gland cancer, biliary tract cancer, bladder cancer, brain or central nervous system cancer, bronchus cancer, blastoma, carcinoma, a chondrosarcoma, cancer of the oral cavity or pharynx, cervical cancer, colon cancer, colorectal cancer, esophageal cancer, gastrointestinal cancer, glioblastoma, hepatic carcinoma, hepatoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, non-small cell lung cancer, ophthalmological cancer, osteosarcoma, ovarian cancer, pancreas cancer, peripheral nervous system cancer, prostate cancer, sarcoma, salivary gland cancer, small bowel or appendix cancer, small-cell lung cancer, squamous cell cancer, stomach cancer, testis cancer, thyroid cancer, urinary bladder cancer, uterine or endometrial cancer, vulval cancer, or any combination thereof.

92. The method of claim 91, wherein the lymphoma is selected from a B-cell lymphoma or a T-cell lymphoma.

93. The method of claim 92, wherein the B-cell or T-cell lymphoma is selected from the group consisting of diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma, intravascular large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma, mantle cell lymphoma, marginal zone B-cell lymphomas, extranodal marginal B-cell lymphomas, mucosa-associated lymphoid tissue (MALT) lymphomas, modal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, Waldenstrom’s macroglobulinemia, hairy cell leukemia, primary central nervous system (CNS) lymphoma, precursor T-lymphoblastic lymphoma/leukemia, peripheral T-cell lymphoma, cutaneous T-cell lymphoma, smoldering adult T-cell lymphoma, chronic adult T-cell lymphoma, acute adult T-cell lymphoma, lymphomatous adult T-cell lymphoma, angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma nasal type (ENKL), enteropathy-associated intestinal T-cell lymphoma (EATL), and anaplastic large cell lymphoma (ALCL).

94. The method of claim 88, wherein the ophthalmological disease is selected from the group consisting of macular degeneration, age-related macular degeneration, diabetes retinopathy, histoplasmosis, macular hole, macular pucker, Bietti’s crystalline dystrophy, retinal detachment, retinal thinning, retinoblastoma, retinopathy of prematurity, Usher’s syndrome, vitreous detachment, Refsum disease, retinitis pigmentosa, onchocerciasis, choroideremia, Leber congenital amaurosis, retinoschisis, juvenile retinoschisis, Stargardt disease, ophthalmoplegia, or any combination thereof.

95. The method of claim 88, wherein the viral infection is caused by a virus selected from the group consisting of West Nile virus, respiratory syncytial virus (RSV), herpes simplex virus 1, herpes simplex virus 2, Epstein-Barr virus (EBV), hepatitis virus A, hepatitis virus B, hepatitis virus C, influenza viruses, chicken pox, avian flu viruses, smallpox, polio viruses, HIV-1, HIV- 2, Ebola virus, and any combination thereof.

96. The method of any one of claims 88-95, wherein the subject is a mammal.

97. The method of claim 96, wherein the subject is human.

98. The method of any one of claims 88-97, further comprising the step of diagnosing the subject with the neurodegenerative disease or disorder, musculoskeletal disease or disorder, cancer, ophthalmological disease or disorder, or viral infection prior to onset of said

administration.

99. The method of any one of claims 88-98, wherein pathology of said neurodegenerative disease or disorder, said musculoskeletal disease or disorder, said cancer, said ophthalmological disease or disorder, or said viral infection comprises stress granules.

100. The method of any one of claims 88-99, wherein pathology of said neurodegenerative disease, said musculoskeletal disease or disorder, said cancer, said ophthalmological disease or disorder, or said viral infection comprises TDP-43 inclusions.

Description:
COMPOUNDS. COMPOSITIONS AND METHODS OF USE

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 62/775,006, filed December 4, 2018, and U.S. Provisional Application No. 62/874,674, filed July 16, 2019, each of which is hereby incorporated in their entirety by reference.

FIELD OF THE INVENTION

The invention relates to compounds, compositions and methods for modulating inclusion formation and stress granules in cells, and for treatment of neurodegenerative diseases, musculoskeletal diseases, cancer, ophthalmological diseases, and viral infections.

BACKGROUND OF THE INVENTION

One of the hallmarks of many neurodegenerative diseases is the accumulation of protein inclusions in the brain and central nervous system. These inclusions are insoluble aggregates of proteins and other cellular components that cause damage to cells and result in impaired function. Proteins such as tau, a-synuclein, huntingtin and b-amyloid have all been found to form inclusions in the brain and are linked to the development of a number of neurodegenerative diseases, including Alzheimer’s disease and Huntington’s disease. Recently, the TDP-43 protein was identified as one of the major components of protein inclusions that typify the

neurogenerative diseases Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Lobar Dementia with ubiquitin inclusions (FTLD-U) (Ash, P.E., et al. (2010) Hum Mol Genet 19(16):3206-3218; Hanson, K.A., et al. (2010) J Biol Chem 285: 11068-11072; Li, Y„ et al. (2010) Proc Natl Acad Sci U.S.A. 107(7):3169-3174; Neumann, M., et al. (2006) Science 314: 130-133; Tsai, K.J., et al. (2010) J Exp Med 207: 1661-1673; Wils, H„ et al. (2010) Proc Natl Acad Sci U.S.A. 170:3858-3863). Abnormalities in TDP-43 biology appear to be sufficient to cause neurodegenerative disease, as studies have indicated that mutations in TDP-43 occur in familial ALS (Barmada, S.J., et al. (2010) J Neurosci 30:639-649; Gitcho, M.A., et al. (2008) Ann Neurol 63(4): 535-538; Johnson, B.S., et al. (2009) J Biol Chem 284:20329-20339; Ling, S.C., et al. (2010) Proc Natl Acad Sci U.S.A. 107:13318-13323; Sreedharan, J., et al. (2008) Science 319: 1668-1672). In addition, TDP-43 has been found to play a role in the stress granule machinery (Colombrita, C., et al. (2009) J Neurochem 111(4): 1051-1061 ; Liu-Yesucevitz, L., et al. (2010) PLoS One 5(10):el3250). Analysis of the biology of the major proteins that accumulate in other neurodegenerative diseases has lead to major advances in our understanding of the pathophysiology of TDP-43 inclusions as well as the development of new drug discovery platforms.

Currently, it is believed that aggregates that accumulate in neurodegenerative diseases like ALS, FTLD-U, Parkinson's disease and Huntington's disease accumulate slowly and are very difficult to disaggregate or perhaps can't be disaggregated. Thus, there is a need in the art for compostions and methods that can rapidly disaggregate these accumulating proteins, more specifically, TDP-43 and/or inhibit the formation of aggregates altogether.

SUMMARY OF THE INVENTION

In one aspect, the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound of Formula (I):

Formula (I) or a pharmaceutically acceptable salt thereof, wherein each of the variables and subvariables thereof are described herein, for example, in the Detailed Description below.

In another aspect, the invention provides a compound of Formula (II):

Formula (II) or a pharmaceutically acceptable salt thereof, wherein each of the variables and subvariables thereof are described herein, for example, in the Detailed Description below.

In embodiments, a compound of Formula (I) or Formula (II) is formulated as a composition (e.g., a pharmaceutical composition).

In another aspect, the invention provides methods for treatment of a neurodegenerative disease or disorder, a musculoskeletal disease or disorder, a cancer, an ophthalmological disease or disorder (e.g., a retinal disease or disorder), and/or a viral infection in a subject, the method comprising administering a compound of Formula (I) or (II) to a subject in need thereof.

In another aspect, the invention provides methods of diagnosing a neurodegenerative disease or disorder, a musculoskeletal disease or disorder, a cancer, an ophthalmological disease or disorder (e.g., a retinal disease or disorder), and/or a viral infection in a subject, the method comprising administering a compound of Formula (I) or (II) to a subject. For use in diagnosis, the compound of Formula (I) or (II) can be modified with a label.

In another aspect, the invention provides methods of modulating stress granules comprising administering a compound of Formula (I) or (II) to a cell or a subject in need thereof. In embodiments, the subject has a neurodegenerative disease or disorder, a musculoskeletal disease or disorder, a cancer, an ophthalmological disease or disorder (e.g., a retinal disease or disorder), and/or a viral infection.

In another aspect, the invention provides methods of modulating TDP-43 inclusion formation comprising administering a compound of Formula (I) or (II) to a cell or a subject in need thereof. In embodiments, the subject has a neurodegenerative disease or disorder, a musculoskeletal disease or disorder, a cancer, an ophthalmological disease or disorder (e.g., a retinal disease or disorder), and/or a viral infection.

In another aspect, the invention provides a method of screening for modulators of TDP- 43 aggregation comprising contacting a compound of Formula (I) or (II) with the cell that expresses TDP-43 and develops spontaneous inclusions.

Still other objects and advantages of the invention will become apparent to those of skill in the art from the disclosure herein, which is simply illustrative and not restrictive. Thus, other embodiments will be recognized by the skilled artisan without departing from the spirit and scope of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1TT are a table of exemplary compounds of the invention.

DETAILED DESCRIPTION OF THE INVENTION

Amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig’s disease or Charcot disease, is a fatal neurodegenerative disease that occurs with an incidence of approximately 1/100,000 (Mitchell, J.D. and Borasio, G.D., (2007) Lancet 369:2031-41). There is currently no therapy for ALS, and the average survival time of patients from the onset of the disease is roughly four years. ALS presents with motor weakness in the distal limbs that rapidly progresses proximally (Mitchell, J.D. and Borasio, G.D., (2007) Lancet 369:2031-41; Lambrechts, D.E., et al. (2004) Trends Mol Med 10:275-282). Studies over the past decade have indicated that TDP- 43 is the major protein that accumulates in affected motor neurons in sporadic ALS (Neumann,

M., et al. (2006) Science 314:130-133). The causes of sporadic ALS are not known, but identification of the major pathological species accumulating in the spinal cord of ALS patients represents a seminal advance for ALS research. To date, TDP-43 is the only protein that has been both genetically and pathologically linked with sporadic ALS, which represents the predominant form of the disease. Multiple papers have identified mutations in TDP-43 associated with sporadic and familial ALS (Sreedharan, J., et al. (2008) Science 319:1668-1672; Gitcho, M.A., et al. (2008) Ann Neurol 63(4):535-538; Neumann, M., et al. (2006) Science 314:130-133). Inhibitors of cell death and inclusions linked to TDP-43 represent a novel therapeutic approach to ALS, and may also elucidate the biochemical pathway linked to the formation of TDP-43 inclusions (Boyd, J.B., et al. (2014) J Biomol Screen 19(l):44-56). As such, TDP-43 represents one of the most promising targets for pharmacotherapy of ALS.

TDP-43 is a nuclear RNA binding protein that translocates to the cytoplasm in times of cellular stress, where it forms cytoplasmic inclusions. These inclusions then colocalize with reversible protein-mRNA aggregates termed“stress granules” (SGs) (Anderson P. and Kedersha,

N. (2008) Trends Biochem Sci 33:141-150; Kedersha, N. and Anderson, P. (2002) Biochem Soc Trans 30:963-969; Lagier-Tourenne, C., et al. (2010) Hum Mol Genet 19:R46-R64). Under many stress-inducing conditions (e.g., arsenite treatment, nutrient deprivation), TDP-43 can co localize with SGs. The reversible nature of SG-based aggregation offers a biological pathway that might be applied to reverse the pathology and toxicity associated with TDP-43 inclusion formation. Studies show that agents that inhibit SG formation also inhibit formation of TDP-43 inclusions (Liu-Yesucevitz, L., et al. (2010) PLoS One 5(10):el3250). The relationship between TDP-43 and stress granules is important because it provides a novel approach for dispersing TDP-43 inclusions using physiological pathways that normally regulate this reversible SG process.. Investigating the particular elements of the SG pathway that regulate TDP-43 inclusion formation can identify selective approaches for therapeutic intervention to delay or halt the progression of disease. Stress granule biology also regulates autophagy and apoptosis, both of which are linked to neurodegeneration. Hence, compounds inhibiting TDP-43 aggregation may play a role in inhibiting neurodegeneration. Compounds

Accordingly, in one aspect, the invention provides a pharmaceutical composition including a pharmaceutically acceptable carrier and compound of Formula (I):

Formula (I)

or a pharmaceutically acceptable salt thereof, wherein:

Ring A is aryl or heteroaryl, optionally substituted with 1-5 R 9 ;

R 1 is H, Ci-Ce alkyl or Ci -O, haloalkyl;

R 2 is H, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -OR B , -C(0)R D , -C(0)OR B , -NR A R C -NR A C(0)R D , - S(0) x R E , -OS(0) x R E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ;

each of R 3 , R 4 , and R 5 is independently H, C1-C6 alkyl, Ci-Ce alkenyl, Ci-Ce alkynyl, Ci- Ce heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -OR B , -C(0)R D , - C(0)OR B , - N R L R c - N R A C (O ) R 13 , -S(0) x R E , -OS(0) x R E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ;

R 6 is C1-C6 alkyl optionally substituted with 1-4 R 8 ;

R 7 is C1-C6 alkyl or -N(R 10 )-CI-C6 alkyl, each of which is optionally substituted with 1-4 or R 6 and R 7 , taken together with the atoms to which they are attached, form a 5-7 membered heterocyclic ring, optionally substituted with 1-5 R 8 , wherein said heterocyclic ring either includes no heteroatoms other than the N and S to which R 6 and R 7 are attached, or includes one additional N ring atom substituted with R 10 ;

each R 8 is independently halo, Ci-Ce alkyl, -OR B , -C(0)0R B , or Ci-Ce haloalkyl;

each R 9 is independently Ci-Ce alkyl, Ci-Ce alkenyl, Ci-Ce alkynyl, Ci-Ce heteroalkyl, Ci-Ce haloalkyl, halo, cyano, nitro, azido, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR B , - C(0)R D , -C(0)0R B , -NR A R C -NR A C(0)R D , -S(0) x R E , -0S(0) x R E , -C(0)NR A S(0) x R E , - NR A S(0) x R E , or -S(0) x NR A ; wherein each cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted by 1-5 R 11 ;

or two R 9 groups bound to adjacent ring atoms of Ring A, taken together with the atoms to which they are attached, form a carbocyclic or heterocyclic ring with Ring A (e.g., a 5-7 membered ring, e.g., a cyclopentyl ring fused to Ring A or a piperidinyl ring fused to Ring A) each R 10 is independently H, Ci-Ce alkyl or Ci-Ce haloalkyl; each R 11 is independently G-G alkyl, G-G alkenyl, G-G alkynyl, G -G heteroalkyl, G-G haloalkyl, halo, cyano, nitro, azido, oxo, cycloalkyl, -OR B , -C(0)R D , -C(0)OR B , -NR A R C -NR A C(0)R D , -S(0)xR E , -OS(0)xR E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A

each R A , R B , R c , R D , or R E is independently H, G-G alkyl, C i -G heteroalkyl, C i -G haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1-4 R 8 ;

or R A and R c , together with the atoms to which each is attached, form a heterocyclyl ring optionally substituted with 1-4 R 8 ; and

x is 0, 1, or 2.

In some embodiments, the compound of Formula (I) is:

Formula (I)

or a pharmaceutically acceptable salt thereof, wherein:

Ring A is aryl or heteroaryl, optionally substituted with 1-5 R 9 ;

R 1 is H, G-G alkyl or G -G haloalkyl;

R 2 is G-G alkyl, G-G alkenyl, G-G alkynyl, G-G heteroalkyl, G-G haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -OR B , -C(0)R D , -C(0)OR B , -NR A R C -NR A C(0)R D , -S(0) x R E , - OS(0) x R E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ;

each of R 3 , R 4 , and R 5 is independently H, G-G alkyl, G-G alkenyl, G-G alkynyl, G- G heteroalkyl, C i -G, haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -OR B , -C(0)R D , - C(0)OR B , -NR A R c , -NR A C(0)R D , -S(0) x R E , -OS(0) x R E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ;

R 6 is G-G alkyl optionally substituted with 1-4 R 8 ;

R 7 is G-G alkyl or -N(R 10 )-C I -C 6 alkyl, each of which is optionally substituted with 1-4 or R 6 and R 7 , taken together with the atoms to which they are attached, form a 5-7 membered heterocyclic ring, optionally substituted with 1-5 R 8 , wherein said heterocyclic ring either includes no heteroatoms other than the N and S to which R 6 and R 7 are attached, or includes one additional N ring atom substituted with R 10 ;

each R 8 is independently halo, G-G alkyl, -OR B , -C(0)OR B , or C i -G, haloalkyl; each R 9 is independently C1-C6 alkyl, CVO, alkenyl, C G, alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR B , - C(0)R D , -C(0)OR B , -NR A R C -NR A C(0)R D , -S(0) x R E , -OS(0) x R E , -C(0)NR A S(0) x R E , - NR A S(0) x R E , or -S(0) x NR A ; wherein each cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted by 1-5 R 11 ;

or two R 9 groups bound to adjacent ring atoms of Ring A, taken together with the atoms to which they are attached, form a carbocyclic or heterocyclic ring with Ring A (e.g., a 5-7 membered ring, e.g., a cyclopentyl ring fused to Ring A or a piperidinyl ring fused to Ring A). each R 10 is independently H, C1-C6 alkyl or C1-C6 haloalkyl;

each R 11 is independently C1-C6 alkyl, Ci-Ce alkenyl, Ci-Ce alkynyl, C1-C6 heteroalkyl,

C1-C6 haloalkyl, halo, cyano, nitro, azido, oxo, cycloalkyl, -OR B , -C(0)R D , -C(0)0R B , -NR A R C , -NR A C(0)R D , -S(0)xR E , -0S(0)xR E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A

each R A , R B , R c , R D , or R E is independently H, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1-4 R 8 ;

or R A and R c , together with the atoms to which each is attached, form a heterocyclyl ring optionally substituted with 1-4 R 8 ; and

x is 0, 1, or 2.

In some embodiments, the moiety is

m is 0, 1, 2, 3, or 4. In some embodiments the moeity is

In some embodiments m is 0.

In some embodiments Ring A is aryl (e.g., phenyl). In some embodiments Ring A is

phenyl. In some embodiments Ring

In some embodiments at least one R 9 group is Ci-Ce alkyl or halo (e.g., chloro). In some embodiments at least one R 9 group is methyl or chloro. In some embodiments one R 9 is chloro or methyl and the other R 9 is C i -G alkyl, G-G alkenyl, G-G alkynyl, C i -G alkoxy, C i -G haloalkyl, halo, cyano, heterocyclyl, phenyl, or heteroaryl; wherein each heterocyclyl, phenyl, or heteroaryl is optionally substituted by 1-5 R 11 .

In some embodiments one R 9 is chloro and the other R 9 is phenyl, monocyclic 4-7 membered heterocyclyl (wherein 1-2 of the ring atoms are heteroatoms selected from the group consisting of N, O, and S), or monocyclic 5-6 membered heteroaryl (wherein 1-2 of the ring atoms are heteroatoms selected from the group consisting of N, O, and S); wherein the phenyl, heterocyclyl, or heteroaryl is optionally substituted with 1-3 R 11 .

In some embodiments, one R 9 group is chloro and the other R 9 group is selected from the group consisting of

wherein n is 0, 1, 2, 3, 4, or 5.

In some embodiments, one R 9 group is chloro and the other R 9 group is selected from the group consisting of Ci-Ce alkyl, G-G alkenyl, G-G heteroalkyl, Ci-Ce haloalkyl, halo, cyano, - OR B , -C(0)OR B , -NR A R C

wherein n is 0, 1, 2, 3, 4, or 5.

In some embodiments n is 0. l R 9

In some embodiments Ring A is V

In some embodiments R 9 is halo (e.g., chloro). In some embodiments R 9 is Ci-Ce alkyl,

Ci-Ce alkenyl, Ci-Ce alkynyl, Ci-Ce heteroalkyl, Ci-Ce haloalkyl, halo, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, or -OR B ; wherein each cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted by 1-5 R 11 . In some embodiments R 9 is C 1 -C 6 alkyl, Ci-Ce alkenyl, C 2 - Ce alkynyl, C 1 -C6 heteroalkyl, phenyl, C 1 -C6 haloalkyl, halo, cyano, or C 1 -C6 alkoxy.

In some embodiments Ring A is heteroaryl. In some embodiments Ring A is monocyclic 5-6 membered heteroaryl wherein 1-2 of the ring atoms are heteroatoms selected from O, N, and S. In some embodiments Ring A is monocyclic 5-6 membered heteroaryl wherein 1-2 of the ring atoms are nitrogen heteroatoms.

In some embodiments Ring A is seleted from the group consisting of

In some embodiments p is 0 or 1.

In some embodiments each R 9 is independently halo, C 1 -C 6 alkyl, C O, alkenyl, C G, alkynyl, C 1 -C6 heteroalkyl, C 1 -C6 haloalkyl, halo, cyano, or C 1 -C6 alkoxy.

In some embodiments Ring A is seleted from the group consisting of

In some embodiments R 1 is H.

In some embodiments R 2 is halo, C 1 -C 6 alkyl or C 1 -C 6 haloalkyl. In some embodiments R 2 is halo. In some embodiments R 2 is chloro.

In some embodiments R 3 is H or halo (e.g., chloro). In some embodiments R 4 and R 5 are both H.

In some embodiments R 4 and R 5 are each H, R 3 is either H or chloro, and R 2 is chloro.

In some embodiments the compound of Formula (I) is a compound of Formula (I-a):

Formula (I-a)

or a pharmaceutically acceptable salt thereof, wherein Ring A, R^R 11 , n, x, p, and subvariables thereof are as described in any of the preceding embodiments, and

wherein W is -NH-, -CH 2 CH 2 -, or -CH 2 NH-, wherein the NH of -CH 2 NH- is linked to the sulfur atom of Formula (I-a).

In some embodiments the compound of Formula (I) is a compound of Formula (I-b):

Formula (I-b)

or a pharmaceutically acceptable salt thereof, wherein R '-R 7 , R 3 ,R 9 -R n , n, x, p, and subvariables thereof are as described in any of the preceding embodiments, and wherein W is - CH 2 -, -CH 2 CH 2 -, or -CH 2 NH-, wherein the NH of -CH 2 NH- is linked to the sulfur atom of Formula (I-b).

In some embodiments the compound of Formula (I) is a compound of Formula (I-c):

Formula (I-c)

or a pharmaceutically acceptable salt thereof, wherein R 2 , R 3 , R 9 -R n , x, p and subvariables thereof are as described in any of the preceding embodiments. In some embodiments the compound of Formula (I) is a compound of Formula (I-d):

Formula (I-d)

or a pharmaceutically acceptable salt thereof, wherein wherein R 2 , R 3 , R 9 -R n , x, p, and subvariables thereof are as described in any of the preceding embodiments.

In some embodiments the compound of Formula (I) is a compound of Formula (I-e):

Formula (I-e)

or a pharmaceutically acceptable salt thereof, wherein R 3 , R 9 -R n , x, p, and subvariables thereof are as described in any of the preceding embodiments.

In some embodiments the compound of Formula (I) is a compound of Formula (I-f):

Formula (I-f)

or a pharmaceutically acceptable salt thereof, wherein R 9 -R n , x, p, and subvariables thereof are as described in any of the preceding embodiments, and wherein R 3 is H or chloro.

In some embodiments the compound of Formula (I) is selected from a compound described in FIGS. 1A-1TT.

In another aspect, the invention provides a compound of Formula (II):

Formula (II)

or a pharmaceutically acceptable salt thereof, wherein:

W is -CH 2 -, -CH2CH2-, -N(R 10 )-, or -CH 2 N(R 10 )-, wherein the N of -CH 2 N(R 10 )- is linked to the sulfur atom of Formula (II);

R 1 is H, C1-C6 alkyl or C1-C6 haloalkyl;

R 2 is H, C1-C6 alkyl, Ci-Ce alkenyl, Ci-Ce alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -OR B , -C(0)R D , -C(0)0R B , -NR A R C , -NR A C(0)R D , - S(0) x R E , -0S(0) x R E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ;

each of R 3 , R 4 , and R 5 is independently H, C1-C6 alkyl, G-G alkenyl, Ci-Ce alkynyl, Ci- Ce heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -OR B , -C(0)R D , - C(0)0R B , -NR A R c , -NR A C(0)R D , -S(0) x R E , -0S(0) x R E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ; or

each R 8 is independently halo, C1-C6 alkyl, -OR B , -C(0)0R B , or C1-C6 haloalkyl;

each R 9 is independently C1-C6 alkyl, Ci-Ce alkenyl, CVG, alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR B , - C(0)R D , -C(0)0R B , -NR A R C , -NR A C(0)R D , -S(0) x R E , -0S(0) x R E , -C(0)NR A S(0) x R E , - NR A S(0) x R E , or -S(0) x NR A ; wherein each cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted by 1-5 R 11 ;

each R 10 is independently H, C1-C6 alkyl or C1-C6 haloalkyl;

each R 11 is independently C1-C6 alkyl, G-G alkenyl, G-G alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, oxo, cycloalkyl, -OR B , -C(0)R D , -C(0)0R B , -NR A R C , -NR A C(0)R D , -S(0) x R E , -0S(0) x R E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ;

each R 12 is independently C1-C6 alkyl, G-G alkenyl, G-G alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -OR B , -C(0)R D , -C(0)0R B , -NR A R C , - NR A C(0)R D , -S(0) x R E , -0S(0) x R E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ;

each R A , R B , R c , R D , or R E is independently H, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1-4 R 8 ; or R A and R c , together with the atoms to which each is attached, form a heterocyclyl ring optionally substituted with 1-4 R 8 ;

x is 0, 1, or 2;

m is 0, 1, 2, 3, or 4;

q is 0, 1, 2, or 3: and

wherein the compound is not

In some embodiments, the invention provides a compound of Formula (II):

Formula (II)

or a pharmaceutically acceptable salt thereof, wherein:

W is -CH 2 -, -CH2CH2-, -N(R 10 )-, or -CH 2 N(R 10 )-, wherein the N of -CH 2 N(R 10 )- is linked to the sulfur atom of Formula (II);

R 1 is FI, C1-C6 alkyl or C1-C6 haloalkyl;

R 2 is C1-C6 alkyl, CVG alkenyl, CVG alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -OR B , -C(0)R D , -C(0)OR B , -NR A R C -NR A C(0)R D , -S(0) x R E , - OS(0)xR E , -C(0)NR A S(0)xR E , -NR A S(0) x R E , or -S(0) x NR A ;

each of R 3 , R 4 , and R 5 is independently FI, C1-C6 alkyl, CVG alkenyl, C2-C6 alkynyl, Ci- Ce heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -OR B , -C(0)R D , - C(0)OR B , -NR A R c , -NR A C(0)R D , -S(0) x R E , -OS(0) x R E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ; or

each R 8 is independently halo, C1-C6 alkyl, -OR B , -C(0)OR B , or C1-C6 haloalkyl;

each R 9 is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR B , - C(0)R D , -C(0)OR B , -NR A R C , -NR A C(0)R D , -S(0) x R E , -OS(0) x R E , -C(0)NR A S(0) x R E , - NR A S(0) x R E , or -S(0) x NR A ; wherein each cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted by 1-5 R 11 ;

each R 10 is independently H, C1-C6 alkyl or C1-C6 haloalkyl;

each R 11 is independently C1-C6 alkyl, G-G alkenyl, G-G alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, oxo, cycloalkyl, -OR B , -C(0)R D , -C(0)0R B , -NR A R C , -NR A C(0)R D , -S(0)xR E , -0S(0)xR E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ;

each R 12 is independently C1-C6 alkyl, C G, alkenyl, G-G alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, nitro, azido, cycloalkyl, -OR B , -C(0)R D , -C(0)0R B , -NR A R C , - NR A C(0)R D , -S(0)xR E , -0S(0)xR E , -C(0)NR A S(0) x R E , -NR A S(0) x R E , or -S(0) x NR A ;

each R A , R B , R c , R D , or R E is independently H, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1-4 R 8 ;

or R A and R c , together with the atoms to which each is attached, form a heterocyclyl ring optionally substituted with 1-4 R 8 ;

x is 0, 1, or 2;

m is 0, 1, 2, 3, or 4;

q is 0, 1, 2, or 3: and

wherein the compound is not

In some embodiments W is -CH2-, -N(R 10 )-, or -CH2N(R 10 )-, wherein the N of - CH2N(R 10 )- is linked to the sulfur atom of Formula (II). In some embodiments, W is -CH2- or - CH2CH2-, and m is 0, 1, 2, 3, or 4. In some embodiments W is -CH2-, and m is 0.

In some embodiments m is 0.

In some embodiments each R 12 is independently C1-C6 alkyl, G-G alkenyl, G-G alkynyl, G-G heteroalkyl, G-G haloalkyl, G-G alkoxy, halo, cyano, or cycloalkyl,

In some embodiments q is 0. In some embodiments at least one R 9 group is halo or CVO, alkyl. In some embodiments at least one R 9 group is chloro or methyl. In some embodiments at least one R 9 group is halo (e.g., chloro). In some embodiments one R 9 is chloro or methyl and the other R 9 is Ci-Ce alkyl, CVO, alkenyl, CVO, alkynyl, C i -O, alkoxy, C i -O, haloalkyl, halo, cyano, heterocyclyl, phenyl, or heteroaryl; wherein each heterocyclyl, phenyl, or heteroaryl is optionally substituted by 1-5 R 11 . In some embodiments one R 9 is chloro or methyl and the other R 9 is phenyl, monocyclic 4- 7 membered heterocyclyl (wherein 1-2 of the ring atoms are heteroatoms selected from the group consisting of N, O, and S), or monocyclic 5-6 membered heteroaryl (wherein 1-2 of the ring atoms are heteroatoms selected from the group consisting of N, O, and S); wherein the phenyl, heterocyclyl, or heteroaryl is optionally substituted with 1-3 R 11 .

In some embodiments one R 9 group is chloro or methyl and the other R 9 group is selected from the group consisting of

wherein n is 0, 1, 2, 3, 4, or 5.

In some embodiments, one R 9 group is chloro and the other R 9 group is selected from the group consisting of C i -O, alkyl, C O, alkenyl, CVO, heteroalkyl, C i -O, haloalkyl, halo, cyano, - OR B , -C(0)0R B , -NR A R C

wherein n is 0, 1, 2, 3, 4, or 5.

In some embodiments n is 0.

In some embodiments R 1 is H.

In some embodiments R 2 is halo, CVG, alkyl or G-G haloalkyl. In some embodiments R 2 is halo. In some embodiments R 2 is chloro.

In some embodiments R 3 is H or halo. In some embodiments R 3 is H or chloro.

In seom embodiments R 4 and R 5 are both H.

In some embodiments R 4 and R 5 are each H, R 3 is H or chloro, and R 2 is chloro.

In some embodiments the compound of Formula (II) is a compound of Formula (Il-a):

Formula (II- a)

or a pharmaceutically acceptable salt thereof, wherein wherein R 2 , R 3 , R 9 -R n , x, and subvariables thereof are as described in any of the preceding embodiments. In some embodiments the compound of Formula (II) is a compound of Formula (Il-b):

Formula (Il-b)

or a pharmaceutically acceptable salt thereof, wherein R 3 , R 9 -R n , x, and subvariables thereof are as described in any of the preceding embodiments.

In some embodiments the compound of Formula (II) is a compound of Formula (II-c):

Formula (II-c)

or a pharmaceutically acceptable salt thereof, wherein R 3 , R 9 -R n , x, and subvariables thereof are as described in any of the preceding embodiments.

In some embodiments the compound of Formula (II) is selected from a compound described in FIGS. 1A-1TT.

In some embodiments the compound of Formula (II) is selected from the group consisting of:

Deuterated Compounds

In some embodiments, compounds described herein (e.g., some compounds of Formula (I) or (II)) are deuterium-enriched. Deuterium (D or 2 H) is a stable, non-radioactive isotope of hydrogen and has an atomic weight of 2.0144. Hydrogen naturally occurs as a mixture of the isotopes 1 H (hydrogen or protium), D ( 2 H or deuterium), and T ( 3 H or tritium). The natural abundance of deuterium is 0.015%. One of ordinary skill in the art recognizes that in all chemical compounds with a H atom, the H atom actually represents a mixture of H and D, with about 0.015% being D. Thus, compounds with a level of deuterium that has been enriched to be greater than its natural abundance of 0.015% should be considered unnatural and, as a result, novel over their non-enriched counterparts.

The effects of deuterium modification on a compound’ s metabolic properties are not predictable, even when deuterium atoms are incorporated at known sites of metabolism. Only by actually preparing and testing a deuterated compound can one determine if and how the rate of metabolism will differ from that of its non-deuterated counterpart. See, for example, Fukuto et al. (J. Med. Chem. 1991, 34, 2871-76). Many compounds have multiple sites where metabolism is possible. The site(s) where deuterium substitution is required and the extent of deuteration necessary to see an effect on metabolism, if any, will be different for each compound.

Unless otherwise stated, when a position is designated specifically as“H” or“hydrogen,” the position is understood to have hydrogen at its natural abundance isotopic composition. Also unless otherwise stated, when a position is designated specifically as“D” or“deuterium,” the position is understood to have deuterium at an abundance that is at least 3000 times greater than the natural abundance of deuterium, which is 0.015% (i.e., the term”D” or“deuterium” indicates at least 45% incorporation of deuterium).

The term“isotopic enrichment factor” as used herein means the ratio between the isotopic abundance of D at the specified position in a compound of this invention and the naturally occurring abundance of that isotope.

Increasing the amount of deuterium present in a compound (e.g., a compound of Formula (I) or (II)) is called“deuterium-enrichment,” and such compounds are referred to as“deuterium- enriched” compounds. If not specifically noted, the percentage of enrichment refers to the percentage of deuterium present in the compound.

In other embodiments, a compound of this invention has an isotopic enrichment factor for each deuterium present at a site designated at a potential site of deuteration on the compound of at least 3500 (52.5.% deuterium incorporation), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6633.3 (99.5% deuterium incorporation). It is understood that the isotopic enrichment factor of each deuterium present at a site designated as a site of deuteration is independent of other deuterated sites. For example, if there are two sites of deuteration on a compound one site could be deuterated at 52.5% while the other could be deuterated at 75%. The resulting compound would be considered to be a compound wherein the isotopic enrichment factor is at least 3500 (52.5%).

Because the natural abundance of deuterium is about 0.015%, a small percentage of naturally occurring compounds of Formula (I) or (II) would be expected to have one naturally occurring compound with one deuterium present.

In some embodiments, the compounds of Formula (I) or (II) comprise an amount of deuterium- enrichment that is more than the amount of deuterium-enrichment present in naturally occurring compounds of Formula (I) or (II).

All percentages given for the amount of deuterium present are mole percentages.

It can be difficult in the laboratory to achieve 100% deuteration at any one site of a lab scale amount of compound (e.g., milligram or greater). When 100% deuteration is recited or a deuterium atom is specifically shown in a structure, it is assumed that a small percentage of hydrogen may still be present. Deuterium-enriched can be achieved by either exchanging protons with deuterium or by synthesizing the molecule with enriched starting materials.

Methods of Use

In the following methods, use of a compound of Formula (I) or (II) can also refer to use of a pharmaceutical compistion including a compound of Formula (I) or (II).

In another aspect, the invention provides a method of modulating stress granule formation, the method comprising contacting a cell with a compound of Formula (I) or (II). In some embodiments, stress granule formation is inhibited. In some embodiments, the stress granule is disaggregated. In some embodiments, stress granule formation is stimulated.

In some embodiments, a compound of Formula (I) or (II) inhibits the formation of a stress granule. The compound of Formula (I) or (II) can inhibit the formation of a stress granule by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% (i.e., complete inhibition) relative to a control.

In some embodiments, a compound of Formula (I) or (II) disaggregates a stress granule. The compound of Formula (I) or (II) can disperses or disaggregate a stress granule by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% (i.e., complete dispersal) relative to a control.

In some embodiments, the stress granule comprises tar DNA binding protein-43 (TDP- 43), T-cell intracellular antigen 1 (TIA-1), TIA1 cytotoxic granule-associated RNA binding protein-like 1 (TIAR, TIAL1), GTPase activating protein binding protein 1 (G3BP-1), GTPase activating protein binding protein 2 (G3BP-2), tris tetraprolin (TTP, ZFP36), fused in sarcoma (FUS), or fragile X mental retardation protein (FMRP, FMR1).

In some embodiments, the stress granule comprises tar DNA binding protein-43 (TDP- 43), T-cell intracellular antigen 1 (TIA-1), TIA1 cytotoxic granule-associated RNA binding protein-like 1 (TIAR, TIAL1), GTPase activating protein binding protein 1 (G3BP-1), GTPase activating protein binding protein 2 (G3BP-2), fused in sarcoma (FUS), or fragile X mental retardation protein (FMRP, FMR1).

In some embodiments, the stress granule comprises tar DNA binding protein-43 (TDP- 43), T-cell intracellular antigen 1 (TIA-1), TIA1 cytotoxic granule-associated RNA binding protein-like 1 (TIAR, TIAL1), GTPase activating protein binding protein 1 (G3BP-1), GTPase activating protein binding protein 2 (G3BP-2), or fused in sarcoma (FUS).

In some embodiments, the stress granule comprises tar DNA binding protein-43 (TDP- 43).

In some embodiments, the stress granule comprises T-cell intracellular antigen 1 (TIA-1).

In some embodiments, the stress granule comprises TIA-1 cytotoxic granule-associated RNA binding protein-like 1 (TIAR, TIAL1).

In some embodiments, the stress granule comprises GTPase activating protein binding protein 1 (G3BP-1).

In some embodiments, the stress granule comprises GTPase activating protein binding protein 2 (G3BP-2).

In some embodiments, the stress granule comprises tris tetraprolin (TTP, ZFP36).

In some embodiments, the stress granule comprises fused in sarcoma (FUS).

In some embodiments, the stress granule comprises fragile X mental retardation protein (FMRP, FMR1).

In another aspect, the invention provides a method of modulating TDP-43 inclusion formation, the method comprising contacting a cell with a compound of Formula (I) or (II). In some embodiments, TDP-43 inclusion formation is inhibited. In some embodiments, the TDP-43 inclusion is disaggregated. In some embodiments, TDP-43 inclusion formation is stimulated.

In some embodiments, a compound of Formula (I) or (II) inhibits the formation of a TDP-43 inclusion. The compound of Formula (I) or (II) can inhibit the formation of a TDP-43 inclusion by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% (i.e., complete inhibition) relative to a control.

In some embodiments, a compound of Formula (I) or (II) disaggregates a TDP-43 inclusion. The compound of Formula (I) or (II) can disperses or disaggregate a TDP-43 inclusion by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% (i.e., complete dispersal) relative to a control.

In another aspect, the invention provides a method for treatment of a neurodegenerative disease or disorder, a musculoskeletal disease or disorder, a cancer, an ophthalmological disease or disorder (e.g., a retinal disease or disorder), and/or a viral infection, the method comprising administering an effective amount of a compound of Formula (I) or (II) to a subject in need thereof.

In some embodiments, the methods are performed in a subject suffering from a neurodegenerative disease or disorder, a musculoskeletal disease or disorder, a cancer, an ophthalmological disease or disorder (e.g., a retinal disease or disorder), and/or a viral infection.

In some embodiments, the methods are performed in a subject suffering from a neurodegenerative disease or disorder. In some embodiments, the methods are performed in a subject suffering from a musculoskeletal disease or disorder. In some embodiments, the methods are performed in a subject suffering from a cancer. In some embodiments, the methods are performed in a subject suffering from an ophthalmological disease or disorder (e.g., a retinal disease or disorder). In some embodiments, the methods are performed in a subject suffering from a viral infection or viral infections.

In some embodiments, the methods comprise administering a compound of Formula (I) or (II) to a subject in need thereof. In some embodiments, the subject is a mammal. In some embodiments, the subject is a nematode. In some embodiments, the subject is human.

In some embodiments, the methods further comprise the step of diagnosing the subject with a neurodegenerative disease or disorder, a musculoskeletal disease or disorder, a cancer, an ophthalmological disease or disorder (e.g., a retinal disease or disorder), or a viral infection prior to administration of a compound of Formula (I) or (II). In some embodiments, the methods further comprise the step of diagnosing the subject with a neurodegenerative disease or disorder prior to administration of a compound of Formula (I) or (II).

In some embodiments, the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, frontotemporal dementia (FTD), FTLD-U, FTD caused by mutations in the progranulin protein or tau protein (e.g., progranulin-deficient FTLD), frontotemporal dementia with inclusion body myopathy (IBMPFD), frontotemporal dementia with motor neuron disease, amyotrophic lateral sclerosis (ALS), Huntington’s disease (HD), Huntington’s chorea, prion diseases (e.g., Creutzf eld- Jacob disease, bovine spongiform encephalopathy, Kuru, and scrapie), Lewy Body disease, diffuse Lewy body disease (DLBD), polyglutamine (polyQ)-repeat diseases, trinucleotide repeat diseases, cerebral degenerative diseases, presenile dementia, senile dementia, Parkinsonism linked to chromosome 17 (FTDP- 17), progressive supranuclear palsy (PSP), progressive bulbar palsy (PBP), psuedobulbar palsy, spinal and bulbar muscular atrophy (SBMA), primary lateral sclerosis, Pick's disease, primary progressive aphasia, corticobasal dementia, HIV-associated dementia, Parkinson's disease, Parkinson's disease with dementia, dementia with Lewy bodies, Down's syndrome, multiple system atrophy, spinal muscular atrophy (SMA, e.g., SMA Type I (e.g., Werdnig-Hoffmann disease), SMA Type II, SMA Type III (e.g., Kugelberg-Welander disease), and congenital SMA with arthrogryposis), progressive spinobulbar muscular atrophy (e.g., Kennedy disease), post polio syndrome (PPS), spinocerebellar ataxia, pantothenate kinase-associated neurodegeneration (PANK), spinal degenerative disease/motor neuron degenerative diseases, upper motor neuron disorder, lower motor neuron disorder, age-related disorders and dementias, Hallervorden-Spatz syndrome, cerebral infarction, cerebral trauma, chronic traumatic encephalopathy, transient ischemic attack, Lytigo-bodig (amyotrophic lateral sclerosis-parkinsonism dementia), Guam- Parkinsonism dementia, hippocampal sclerosis, corticobasal degeneration, Alexander disease, Apler’s disease, Krabbe’s disease, neuroborreliosis, neurosyphilis, Sandhoff disease, Tay-Sachs disease, Schilder’s disease, Batten disease, Cockayne syndrome, Kearns-Sayre syndrome, Gerstmann-Straussler-Scheinker syndrome and other transmissible spongiform

encephalopathies, hereditary spastic paraparesis, Leigh’s syndrome, demyelinating diseases, neuronal ceroid lipofuscinoses, epilepsy, tremors, depression, mania, anxiety and anxiety disorders, sleep disorders (e.g., narcolepsy, fatal familial insomnia), acute brain injuries (e.g., stroke, head injury) autism, other diseases or disorders relating to the aberrant expression of TDP-43 and altered proteostasis, and any combination thereof.

In some embodiments, the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, frontotemporal dementia (FTD), FTLD-U, FTD caused by mutations in the progranulin protein or tau protein (e.g., progranulin-deficient FTLD), amyotrophic lateral sclerosis (ALS), Huntington’s disease (HD), Huntington’s chorea, Creutzfeld- Jacob disease, senile dementia, Parkinsonism linked to chromosome 17 (FTDP-17), progressive supranuclear palsy (PSP), Pick's disease, primary progressive aphasia, corticobasal dementia, Parkinson's disease, Parkinson's disease with dementia, dementia with Lewy bodies, Down's syndrome, multiple system atrophy, spinal muscular atrophy (SMA), spinocerebellar ataxia, spinal degenerative disease/motor neuron degenerative diseases, Hallervorden-Spatz syndrome, cerebral infarction, cerebral trauma, chronic traumatic encephalopathy, transient ischemic attack, Lytigo-bodig (amyotrophic lateral sclerosis-parkinsonism dementia), hippocampal sclerosis, corticobasal degeneration, Alexander disease, Cockayne syndrome, and any combination thereof.

In some embodiments, the neurodegenerative disease is frontotemporal dementia (FTD). In some embodiments, the neurodegenerative disease is Alzheimer's disease or amyotrophic lateral sclerosis (ALS).

In some embodiments, the musculoskeletal disease is selected from the group consisting of muscular dystrophy, facioscapulohumeral muscular dystrophy (e.g., FSHD1 or FSHD2), Freidrich’s ataxia, progressive muscular atrophy (PMA), mitochondrial encephalomyopathy (MELAS), multiple sclerosis, inclusion body myopathy, inclusion body myositis (e.g., sporadic inclusion body myositis), post-polio muscular atrophy (PPMA), motor neuron disease, myotonia, myotonic dystrophy, sacropenia, multifocal motor neuropathy, inflammatory myopathies, paralysis, and other diseases or disorders relating to the aberrant expression of TDP-43 and altered proteostasis.

In some embodiments, compounds of Formula (I) or (II) may be used to prevent or treat symptoms caused by or relating to said musculoskeletal diseases, e.g., kyphosis, hypotonia, foot drop, motor dysfunctions, muscle weakness, muscle atrophy, neuron loss, muscle cramps, altered or aberrant gait, dystonias, astrocytosis (e.g., astrocytosis in the spinal cords), liver disease, respiratory disease or respiratory failure, inflammation, headache, and pain (e.g., back pain, neck pain, leg pain, or inflammatory pain).

In some embodiments, the cancer is selected from the group consisting of breast cancer, a melanoma, adrenal gland cancer, biliary tract cancer, bladder cancer, brain or central nervous system cancer, bronchus cancer, blastoma, carcinoma, a chondrosarcoma, cancer of the oral cavity or pharynx, cervical cancer, colon cancer, colorectal cancer, esophageal cancer, gastrointestinal cancer, glioblastoma, hepatic carcinoma, hepatoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, non-small cell lung cancer, ophthalmological cancer, osteosarcoma, ovarian cancer, pancreas cancer, peripheral nervous system cancer, prostate cancer, sarcoma, salivary gland cancer, small bowel or appendix cancer, small-cell lung cancer, squamous cell cancer, stomach cancer, testis cancer, thyroid cancer, urinary bladder cancer, uterine or endometrial cancer, vulval cancer, and any combination thereof. In some embodiments, the cancer is selected from the group consisting of blastoma, carcinoma, a glioblastoma, hepatic carcinoma, lymphoma, leukemia, and any combination thereof.

In some embodiments, the cancer is selected from Hodgkin’s lymphoma or non- Hodgkin’s lymphoma. In some embodiments, the cancer is a non-Hodgkin’s lymphoma, selected from the group consisting of a B-cell lymphoma (e.g., diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma, intravascular large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma, mantle cell lymphoma, marginal zone B-cell lymphomas, extranodal marginal B-cell lymphomas, mucosa-associated lymphoid tissue (MALT) lymphomas, modal marginal zone B-cell lymphoma, splenic marginal zone B- cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, Waldenstrom’s

macroglobulinemia, hairy cell leukemia, and primary central nervous system (CNS) lymphoma) and a T-cell lymphoma (e.g., precursor T-lymphoblastic lymphoma/leukemia, peripheral T-cell lymphoma, cutaneous T-cell lymphoma, adult T-cell lymphoma (e.g., smoldering adult T-cell lymphoma, chronic adult T-cell lymphoma, acute adult T-cell lymphoma, lymphomatous adult T-cell lymphoma), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma nasal type (ENKL), enteropathy-associated intestinal T-cell lymphoma (EATL) (e.g., Type I EATL and Type II EATL), and anaplastic large cell lymphoma (ALCL)).

In some embodiments, the ophthalmological disease or disorder (e.g., retinal disease or disorder) is selected from macular degeneration (e.g., age-related macular degeneration), diabetes retinopathy, histoplasmosis, macular hole, macular pucker, Bietti’s crystalline dystrophy, retinal detachment, retinal thinning, retinoblastoma, retinopathy of prematurity, Usher’s syndrome, vitreous detachment, Refsum disease, retinitis pigmentosa, onchocerciasis, choroideremia, Leber congenital amaurosis, retinoschisis (e.g., juvenile retinoschisis), Stargardt disease, ophthalmoplegia, and the like.

In some embodiments, the ophthalmological disease or disorder (e.g., retinal disease or disorder) is selected from macular degeneration (e.g., age-related macular degeneration), diabetes retinopathy, histoplasmosis, macular hole, macular pucker, Bietti’s crystalline dystrophy, retinoblastoma, retinopathy of prematurity, Usher’s syndrome, Refsum disease, retinitis pigmentosa, onchocerciasis, choroideremia, Leber congenital amaurosis, retinoschisis (e.g., juvenile retinoschisis), Stargardt disease, and the like.

In some embodiments, the viral infection is caused by a virus selected from the group consisting of West Nile virus, respiratory syncytial virus (RSV), herpes simplex virus 1, herpes simplex virus 2, Epstein-Barr virus (EBV), hepatitis virus A, hepatitis virus B, hepatitis virus C, influenza viruses, chicken pox, avian flu viruses, smallpox, polio viruses, HIV-1, HIV-2, Ebola virus, and any combination thereof.

In some embodiments, the viral infection is caused by a virus selected from the group consisting of herpes simplex virus 1, herpes simplex virus 2, Epstein-Barr virus (EBV), hepatitis virus A, hepatitis virus B, hepatitis virus C, HIV-1, HIV-2, Ebola virus, and any combination thereof.

In some embodiments, the viral infection is HIV-1 or HIV-2.

In some embodiments, the pathology of the neurodegenerative disease or disorder, musculoskeletal disease or disorder, cancer, ophthalmological disease or disorder (e.g., retinal disease or disorder), and/or viral infection comprises stress granules.

In some embodiments, pathology of the disease or disorder comprises stress granules. By comprising stress granules is meant that number of stress granules in a cell in the subject is changed relative to a control and/or healthy subject or relative to before onset of said disease or disorder. Exemplary diseases and disorders pathology of which incorporate stress granules include, but are not limited to, neurodegenerative diseases, musculoskeletal diseases, cancers, ophthalmological diseases (e.g., retinal diseases), and viral infections.

In another aspect, the invention provides methods of diagnosing a neurodegenerative disease, a musculoskeletal disease, a cancer, an ophthalmological disease (e.g., a retinal disease), or a viral infection in a subject, the method comprising administering a compound of Formula (I) to the subject. In some embodiments, the invention provides methods of diagnosing a neurodegenerative disease in a subject, the method comprising administering a compound of Formula (I) to the subject. For use in diagnosis, a compound of Formula (I) can be modified with a label.

In another aspect, the invention provides methods of modulating stress granules comprising contacting a cell with a compound of Formula (I) or (II).

In another aspect, the invention provides methods of modulating TDP-43 inclusion formation comprising contacting a cell with a compound of Formula (I) or (II). In some embodiments, TDP-43 is inducibly expressed. In some embodiments, the cell line is a neuronal cell line.

In some embodiments, the cell is treated with a physiochemical stressor. In some embodiments, the physicochemical stressor is selected from arsenite, nutrient deprivation, heat shock, osmotic shock, a virus, genotoxic stress, radiation, oxidative stress, oxidative stress, a mitochondrial inhibitor, and an endoplasmic reticular stressor. In some embodiments, the physicochemical stressor is ultraviolet or x-ray radiation. In some embodiments, the physicochemical stressor is oxidative stress induced by FcCb or CuCb and a peroxide.

In yet another aspect, the invention provides a method of screening for modulators of TDP-43 aggregation comprising contacting a compound of Formula (I) or (II) with a cell that expresses TDP-43 and develops spontaneous inclusions.

In some embodiments, the stress granule comprises TDP-43, i.e., is a TDP-43 inclusion. Accordingly, in some embodiments, a compound of Formula (I) or (II) is a modulator of TDP-43 inclusions.

In some embodiments, the subject is a mammal. In some embodiments, the subject is human.

In some embodiments, the method further comprises the step of diagnosing the subject with the neurodegenerative disease or disorder, musculoskeletal disease or disorder, cancer, ophthalmological disease or disorder, or viral infection prior to onset of said administration. In some embodiments, the pathology of said neurodegenerative disease or disorder, said musculoskeletal disease or disorder, said cancer, said ophthalmological disease or disorder, and said viral infection comprises stress granules. In some embodiments, the pathology of said neurodegenerative disease, said musculoskeletal disease or disorder, said cancer, said ophthalmological disease or disorder, and said viral infection comprises TDP-43 inclusions.

TDP-43 and other RNA-binding proteins function in both the nucleus and cytoplasm to process mRNA, e.g., by splicing mRNA, cleaving mRNA introns, cleaving untranslated regions of mRNA or modifying protein translation at the synapse, axon, dendrite or soma. Therefore, targeting other proteins that function in an analogous manner to TDP-43 or by processing mRNA may also be beneficial to prevent and treat neurodegeneration resulting from disease. For instance, the fragile X mental retardation 1 (FMRP) protein is essential for normal cognitive development (Nakamoto, M., et al. (2007) Proc Natl Acad Sci U.S.A. 104:15537-15542). The signaling systems that affect TDP-43 function might also affect this protein, thus improving cognitive function. This can be particularly important at the synapse where neurons

communicate. Without wishing to be bound by a theory, the signaling systems that conpounds of Formula (I) or (II) target may also modify these processes, which play a role in

neurodegeneration or mental health illnesses (e.g., schizophrenia).

The cellular stress response follows a U-shaped curve. Overinduction of this pathway, such as observed in many neurodegenerative diseases, can be harmful for cells. However, a decreased stimulation of this pathway can also be harmful for cells, e.g., in the case of an acute stress, such as a stroke. Thus, the appropriate action for some diseases is the inhibition of stress granule formation, while for other diseases, stimulation of stress granule formation is beneficial.

In some embodiments, the TDP-43 protein in a stress granule may be wild-type or a mutant form of TDP-43. In some embodiments, the mutant form of TDP-43 comprises an amino acid addition, deletion, or substitution, e.g., relative to the wild type sequence of TDP-43. In some embodiments, the mutant form of TDP-43 comprises an amino acid substitution relative to the wild type sequence, e.g., a G294A, A135T, Q331K, or Q343R substitution. In some embodiments, the TDP-43 protein in a stress granule comprises a post-translational modification, e.g., phosphorylation of an amino acid side chain, e.g., T103, S104, S409, or S410. In some embodiments, post-translational modification of the TDP-43 protein in a stress granule may be modulated by treatment with a compound of the invention.

Methods of Treatment

Neurodegenerative diseases: Without wishing to be bound by a theory, compounds of Formula (I) or (II) can be used to delay the progression of neurodegenerative illnesses where the pathology incorporates stress granules. Such illnesses include ALS and frontotemporal dementia, in which TDP-43 is the predominant protein that accumulates to form the pathology. This group also includes Alzheimer’s disease and FTLD-U, where TDP-43 and other stress granule proteins co-localize with tau pathology. Because modulators of TDP-43 inclusions, such as compounds of Formula (I) or (II), can act to block the enzymes that signal stress granule formation (e.g., the three enzymes that phosphorylate eIF2a: PERK, GCN2 and HRI), compounds of Formula (I) or (II) may also reverse stress granules that might not include TDP-43. Accordingly, compounds of Formula (I) or (II) can be used for treatment of neurodegenerative diseases and disorders in which the pathology incorporates stress granules, such as Huntington’s chorea and Creutzfeld- Jacob disease. Compounds of Formula (I) or (II) may also be used for treatment of

neurodegenerative diseases and disorders that involve TDP-43 multisystem proteinopathy.

The term“neurodegenerative disease” as used herein, refers to a neurological disease characterized by loss or degeneration of neurons. The term“neurodegenerative disease” includes diseases caused by the involvement of genetic factors or the cell death (apoptosis) of neurons attributed to abnormal protein accumulation and so on. Additionally, neurodegenerative diseases include neurodegenerative movement disorders and neurodegenerative conditions relating to memory loss and/or dementia. Neurodegenerative diseases include tauopathies and oc- synucleopathies. Exemplary neurodegenerative diseases include, but are not limited to,

Alzheimer's disease, frontotemporal dementia (FTD), FTLD-U, FTD caused by mutations in the progranulin protein or tau protein (e.g., progranulin-deficient FTLD), frontotemporal dementia with inclusion body myopathy (IBMPFD), frontotemporal dementia with motor neuron disease, amyotrophic lateral sclerosis (ALS), amyotrophic lateral sclerosis with dementia (ALSD), Huntington's disease (HD), Huntington’s chorea, prion diseases (e.g., Creutzfeld- Jacob disease, bovine spongiform encephalopathy, Kuru, or scrapie), Lewy Body disease, diffuse Lewy body disease (DLBD), polyglutamine (polyQ)-repeat diseases, trinucleotide repeat diseases, cerebral degenerative diseases, presenile dementia, senile dementia, Parkinsonism linked to chromosome 17 (FTDP-17), progressive supranuclear palsy (PSP), progressive bulbar palsy (PBP), psuedobulbar palsy, spinal and bulbar muscular atrophy (SBMA), primary lateral sclerosis, Pick's disease, primary progressive aphasia, corticobasal dementia, HIV-associated dementia, Parkinson's disease, Parkinson's disease with dementia, dementia with Lewy bodies, Down's syndrome, multiple system atrophy, spinal muscular atrophy (SMA, e.g., SMA Type I (e.g., Werdnig-Hoffmann disease) SMA Type II, SMA Type III (e.g., Kugelberg-Welander disease), and congenital SMA with arthrogryposis), progressive spinobulbar muscular atrophy (e.g., Kennedy disease), post-polio syndrome (PPS), spinocerebellar ataxia, pantothenate kinase- associated neurodegeneration (PANK), spinal degenerative disease/motor neuron degenerative diseases, upper motor neuron disorder, lower motor neuron disorder, age-related disorders and dementias, Hallervorden-Spatz syndrome, Lytigo-bodig (amyotrophic lateral sclerosis- parkinsonism dementia), Guam-Parkinsonism dementia, hippocampal sclerosis, corticobasal degeneration, Alexander disease, Apler’s disease, Krabbe’s disease, neuroborreliosis, neurosyphilis, Sandhoff disease, Schilder’s disease, Batten disease, Cockayne syndrome, Kearns-Sayre syndrome, Gerstmann-Straussler-Scheinker syndrome, hereditary spastic paraparesis, Leigh’s syndrome, demyelinating diseases, epilepsy, tremors, depression, mania, anxiety and anxiety disorders, sleep disorders (e.g., narcolepsy, fatal familial insomnia), acute brain injuries (e.g., stroke, head injury) and autism. As used herein, the term "oc-synucleopathy" refers to a neurodegenerative disorder or disease involving aggregation of oc-synuclein or abnormal oc-synuclein in nerve cells in the brain (Ostrerova, N., et al. (1999) J Neurosci 19:5782:5791 ; Rideout, H.J., et al. (2004) J Biol Chem 279:46915-46920). oc-Synucleopathies include, but are not limited to, Parkinson's disease, Parkinson's disease with dementia, dementia with Lewy bodies, Pick's disease, Down's syndrome, multiple system atrophy, amylotrophic lateral sclerosis (ALS), Hallervorden-Spatz syndrome, and the like.

As used herein, the term“tauopathy” refers to a neurodegenerative disease associated with the pathological aggregation of tau protein in the brain. Tauopathies include, but are not limited to, Alzheimer’ s disease, Pick's disease, corticobasal degeneration, Argyrophilic grain disease (AGD), progressive supranuclear palsy, Frontotemporal dementia, Frontotemporal lobar degeneration, or Pick's complex.

Musculoskeletal diseases : Musculoskeletal diseases and disorders as defined herein are conditions that affect the muscles, ligaments, tendons, and joints, as well as the skeletal structures that support them. Without wishing to be bound by a theory, aberrant expression of certain proteins, such as the full-length isoform of DUX4, has been shown to inhibit protein turnover and increase the expression and aggregation of cytotoxic proteins including insoluble TDP-43 in skeletal muscle cells (Homma, S. et al. Ann Clin Transl Neurol (2015) 2:151-166).

As such, compounds of Formula (I) or (II) may be used to prevent or treat a musculoskeletal disease, e.g., a musculoskeletal disease that results in accumulation of TDP-43 and other stress granule proteins, e.g., in the nucleus, cytoplasm, or cell bodies of a muscle cell or motor neuron. Exemplary musculoskeletal diseases include muscular dystrophy, facioscapulohumeral muscular dystrophy (e.g., FSHD1 or FSHD2), Freidrich’s ataxia, progressive muscular atrophy (PMA), mitochondrial encephalomyopathy (MEL AS), multiple sclerosis, inclusion body myopathy, inclusion body myositis (e.g., sporadic inclusion body myositis), post-polio muscular atrophy (PPMA), motor neuron disease, myotonia, myotonic dystrophy, sacropenia, spasticity, multifocal motor neuropathy, inflammatory myopathies, paralysis, and other diseases or disorders relating to the aberrant expression of TDP-43 and altered proteostasis. In addition, compounds of Formula (I) or (II) may be used to prevent or treat symptoms caused by or relating to said musculoskeletal diseases, e.g., kyphosis, hypotonia, foot drop, motor dysfunctions, muscle weakness, muscle atrophy, neuron loss, muscle cramps, altered or aberrant gait, dystonias, astrocytosis (e.g., astrocytosis in the spinal cords), liver disease, inflammation, headache, pain (e.g., back pain, neck pain, leg pain, inflammatory pain), and the like. In some embodiments, a musculoskeletal disease or a symptom of a musculoskeletal disease may overlap with a neurodegenerative disease or a symptom of a neurodegenerative disease.

Cancers: Cancer cells grow quickly and in low oxygen environments by activating different elements of the cellular stress response. Researchers have shown that drugs targeting different elements of the stress response can be anti-neoplastic. For example, rapamycin blocks mTOR, upregulates autophagy and inhibits some types of tumors. Proteasomal inhibitors, such as velcade (Millenium Pharma) are used to treat some cancers. HSP90 inhibitors, such as 17- allylaminogeldanamycin (17AAG), are currently in clinical trials for cancer. Without wishing to be bound by a theory, compounds of Formula (I) may also be used for treatment of cancer, as a greater understanding of the role of TDP-43 in RNA processing and transcription factor signaling has recently begun to emerge (Lagier-Tourenne, C., et al. (2010) Hum Mol Genet 19:R46-R64; Ayala, Y. M„ et al. (2008) Proc Natl Acad Sci U.S.A. 105(10):3785-3789).

Additionally, TDP-43 modulators can be combined with one or more cancer therapies, such as chemotherapy and radiation therapy.

A "cancer" in a subject refers to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and certain characteristic morphological features. Often, cancer cells will be in the form of a tumor, but such cells may exist alone within an animal, or may be a non-tumorigenic cancer cell, such as a leukemia cell. In some circumstances, cancer cells will be in the form of a tumor; such cells may exist locally within an animal, or circulate in the blood stream as independent cells, for example, leukemic cells. Examples of cancer include but are not limited to breast cancer, a melanoma, adrenal gland cancer, biliary tract cancer, bladder cancer, brain or central nervous system cancer, bronchus cancer, blastoma, carcinoma, a

chondrosarcoma, cancer of the oral cavity or pharynx, cervical cancer, colon cancer, colorectal cancer, esophageal cancer, gastrointestinal cancer, glioblastoma, hepatic carcinoma, hepatoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, non-small cell lung cancer, ophthalmological cancer, osteosarcoma, ovarian cancer, pancreas cancer, peripheral nervous system cancer, prostate cancer, sarcoma, salivary gland cancer, small bowel or appendix cancer, small-cell lung cancer, squamous cell cancer, stomach cancer, testis cancer, thyroid cancer, urinary bladder cancer, uterine or endometrial cancer, vulval cancer, and the like.

Other exemplary cancers include, but are not limited to, ACTH-producing tumors, acute lymphocytic leukemia, acute nonlymphocytic leukemia, cancer of the adrenal cortex, bladder cancer, brain cancer, breast cancer, cervical cancer, chronic lymphocytic leukemia, chronic myelocytic leukemia, colorectal cancer, cutaneous T-cell lymphoma, endometrial cancer, esophageal cancer, Ewing's sarcoma, gallbladder cancer, hairy cell leukemia, head & neck cancer, ophthalmological cancer, Hodgkin's lymphoma, Kaposi's sarcoma, kidney cancer, liver cancer, lung cancer (small and/or non-small cell), malignant peritoneal effusion, malignant pleural effusion, melanoma, mesothelioma, multiple myeloma, neuroblastoma, non-Hodgkin's lymphoma, osteosarcoma, ovarian cancer, ovary (germ cell) cancer, prostate cancer, pancreatic cancer, penile cancer, retinoblastoma, skin cancer, soft-tissue sarcoma, squamous cell carcinomas, stomach cancer, testicular cancer, thyroid cancer, trophoblastic neoplasms, uterine cancer, vaginal cancer, cancer of the vulva, Wilm's tumor, and the like.

Exemplary lymphomas include Hodgkin’s lymphoma and non-Hodgkin’s lymphoma. Further exemplification of non-Hodgkin’s lymphoma include, but are not limited to, B-cell lymphomas (e.g., diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma, intravascular large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma, mantle cell lymphoma, marginal zone B-cell lymphomas, extranodal marginal B-cell lymphomas, mucosa-associated lymphoid tissue (MALT) lymphomas, modal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, Waldenstrom’s macroglobulinemia, hairy cell leukemia, and primary central nervous system (CNS) lymphoma) and T-cell lymphomas (e.g., precursor T- lymphoblastic lymphoma/leukemia, peripheral T-cell lymphoma, cutaneous T-cell lymphoma, adult T-cell lymphoma (e.g., smoldering adult T-cell lymphoma, chronic adult T-cell lymphoma, acute adult T-cell lymphoma, lymphomatous adult T-cell lymphoma), angioimmunoblastic T- cell lymphoma, extranodal natural killer T-cell lymphoma nasal type (ENKL), enteropathy- associated intestinal T-cell lymphoma (EATL) (e.g., Type I EATL and Type II EATL), and anaplastic large cell lymphoma (ALCL)).

Ophthalmological diseases: Ophthalmological diseases and disorders (e.g., retinal diseases and disorders) as defined herein affect the retina and other parts of the eye and may contribute to impaired vision and blindness. Several ophthalmological diseases (e.g., retinal diseases) are characterized by the accumulation of protein inclusions and stress granules within or between cells of the eye, e.g., retinal cells and nearby tissues. In addition, an ophthalmological disease (e.g., retinal disease) may also be a symptom of or precursor to neurogenerative diseases, such as ALS and FTD (Ward, M.E., et al. (2014) J Exp Med 211(10): 1937). Therefore, use of compounds that may inhibit formation of protein inclusions and stress granules, including compounds of Formula (I) or (II), may play an important role in the prevention or treatment of ophthalmological diseases (e.g., retinal diseases).

Exemplary ophthalmological diseases (e.g., retinal diseases) include, but are not limited to, macular degeneration (e.g., age-related macular degeneration), diabetes retinopathy, histoplasmosis, macular hole, macular pucker, Bietti’s crystalline dystrophy, retinal detachment, retinal thinning, retinoblastoma, retinopathy of prematurity, Usher’s syndrome, vitreous detachment, Refsum disease, retinitis pigmentosa, onchocerciasis, choroideremia, Leber congenital amaurosis, retinoschisis (e.g., juvenile retinoschisis), Stargardt disease,

ophthalmoplegia, and the like.

Viral infections: Stress granules often form during viral illnesses, as viral infections often involve hijacking the cellular reproductive machinery toward production of viral proteins. In this case, inhibitors of stress granules can be useful for interfering with viral function. Other viruses appear to inhibit SG formation to prevent the cell from mobilizing a stress response. In such a case, an inducer of stress granules can interfere with viral activity and help combat viral infections (e.g., Salubrinal, an eIF2a phosphatase inhibitor and stress granule inducer). Two viruses for which SG biology has been investigated include West Nile virus and respiratory syncytial virus (RSV) (Emara, M.E. and Brinton, M. A. (2007) Proc. Natl. Acad. Sci. USA 104(21): 9041-9046). Therefore, use of compounds that may inhibit formation of protein inclusions and stress granules, including compounds of Formula (I) or (II), may be useful for the prevention and/or treatment of a viral infection.

Exemplary viruses include, but are not limited to, West Nile virus, respiratory syncytial virus (RSV), Epstein-Barr virus (EBV), hepatitis A, B, C, and D viruses, herpes viruses, influenza viruses, chicken pox, avian flu viruses, smallpox, polio viruses, HIV, Ebola virus, and the like.

Definitions

Unless stated otherwise, or implicit from context, the following terms and phrases include the meanings provided below. Unless explicitly stated otherwise, or apparent from context, the terms and phrases below do not exclude the meaning that the term or phrase has acquired in the art to which it pertains. The definitions are provided to aid in describing particular embodiments, and are not intended to limit the claimed invention, because the scope of the invention is limited only by the claims. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

As used herein, the terms“compounds” and“agent” are used interchangeably to refer to the inhibitors/antagonists/agonists of the invention. In certain embodiments, the compounds are small organic or inorganic molecules, e.g., with molecular weights less than 7500 amu, preferably less than 5000 amu, and even more preferably less than 2000, 1500, 1000, 750, 600, or 500 amu. In certain embodiments, one class of small organic or inorganic molecules are non- peptidyl, e.g., containing 2, 1, or no peptide and/or saccharide linkages.

Unless otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood as modified in all instances by the term“about.” The term“about” when used in connection with percentages may mean ±1%.

The singular terms“a,”“an,” and“the” refer to one or to more than one, unless context clearly indicates otherwise. Similarly, the word“or” is intended to include“and” unless the context clearly indicates otherwise.

Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below. As used herein, the term“administer” refers to the placement of a composition into a subject by a method or route which results in at least partial localization of the composition at a desired site such that desired effect is produced. A compound or composition described herein can be administered by any appropriate route known in the art including, but not limited to, oral or parenteral routes, including intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, nasal, rectal, intrathecal, and topical (including buccal and sublingual) administration.

The terms“decrease”,“reduced”,“reduction” ,“decrease” or“inhibit” are all used herein generally to mean a decrease by a statistically significant amount. In some embodiments, the terms“reduced”,“reduction”,“decrease” or“inhibit” mean a decrease by at least 0.1% as compared to a reference level, for example a decrease by at least about 1%, or at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% decrease (e.g. absent level as compared to a reference sample), or any decrease between 1-100%, e.g., 10-100% as compared to a reference level.

The terms“increased”,’’increase”,“enhance” or“activate” are all used herein to generally mean an increase by a statically significant amount. In some embodiments, the terms “increased”,“increase”,“enhance” or“activate” mean an increase by at least 0.1% as compared to a reference level, for example a decrease by at least about 1%, or at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase (e.g. absent level as compared to a reference sample), or any increase between 1-100%, e.g., 10-100% as compared to a reference level.

By“treatment”,“prevention” or“amelioration” of a disease or disorder is meant delaying or preventing the onset of such a disease or disorder, reversing, alleviating, ameliorating, inhibiting, slowing down or stopping the progression, aggravation or deterioration the progression or severity of a condition associated with such a disease or disorder. In one embodiment, at least one symptom of a disease or disorder is alleviated by at least about 1%, or at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%.

As used herein, an amount of a compound or combination effective to beat a disorder (e.g., a disorder as described herein),“therapeutically effective amount” or“effective amount” refers to an amount of the compound or combination which is effective, upon single or multiple dose administration(s) to a subject, in treating a subject, or in curing, alleviating, relieving or improving a subject with a disorder (e.g., a disorder as described herein) beyond that expected in the absence of such treatment. Determination of a therapeutically effective amount is well within the capability of those skilled in the art. Generally, a therapeutically effective amount can vary with the subject’s history, age, condition, sex, as well as the severity and type of the medical condition in the subject, and administration of other pharmaceutically active agents.

As used herein, a“subject” means a human or animal. Usually the animal is a vertebrate such as a primate, rodent, domestic animal or game animal. Primates include chimpanzees, cynomologous monkeys, spider monkeys, and macaques, e.g., Rhesus. Rodents include mice, rats, woodchucks, ferrets, rabbits and hamsters. Domestic and game animals include cows, horses, pigs, deer, bison, buffalo, feline species, e.g., domestic cat, canine species, e.g., dog, fox, wolf, avian species, e.g., chicken, emu, ostrich, and fish, e.g., trout, catfish and salmon. Patient or subject includes any subset of the foregoing, e.g., all of the above, but excluding one or more groups or species such as humans, primates or rodents. In certain embodiments, the subject is a mammal, e.g., a primate, e.g., a human. The terms,“patient” and“subject” are used

interchangeably herein. The terms,“patient” and“subject” are used interchangeably herein.

The term "nucleic acid" as used herein refers to a polymeric form of nucleotides, either ribonucleotides or deoxynucleotides or a modified form of either type of nucleotide. The terms should also be understood to include, as equivalents, analogs of either RNA or DNA made from nucleotide analogs, and, as applicable to the embodiment being described, single-stranded (such as sense or antisense) and double-stranded polynucleotides.

As used herein, the terms“modulator of stress granule” and“stress granule modulator” refer to compounds and compositions of Formula (I) or (II) that modulate the formation and/or disaggregation of stress granules.

The term“TDP-43 inclusion” as used herein refers to protein aggregates that comprise TDP-43 proteins. The TDP-43 protein in the inclusion can be wild-type or a mutant form of TDP-43.

As used herein, the terms“modulator of TDP-43 inclusion” and“TDP-43 inclusion modulator” refer to compounds and compositions of Formula (I) or (II) that modulate the formation and/or disaggregation of cytoplasmic TDP-43 inclusions. Selected Chemical Definitions

At various places in the present specification, substituents of compounds of the invention are disclosed in groups or in ranges. It is specifically intended that the invention include each and every individual subcombination of the members of such groups and ranges. For example, the term“Ci- 6 alkyl” is specifically intended to individually disclose methyl, ethyl, propyl, butyl, pentyl and hexyl.

For compounds of the invention in which a variable appears more than once, each variable can be a different moiety selected from the Markush group defining the variable. For example, where a structure is described having two R groups that are simultaneously present on the same compound; the two R groups can represent different moieties selected from the Markush group defined for R.

It is further appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, can also be provided in combination in a single embodiment. Conversely, various features of the invention which are, for brevity, described in the context of a single embodiment, can also be provided separately or in any suitable subcombination.

If a compound of the present invention is depicted in the form of a chemical name and as a formula, in case of any discrepancy, the formula shall prevail.

The symbol , whether utilized as a bond or displayed perpendicular to a bond indicates the point at which the displayed moiety is attached to the remainder of the molecule, solid support, etc.

The following terms are intended to have the meanings presented therewith below and are useful in understanding the description and intended scope of the present invention.

As used herein,“alkyl” refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 24 carbon atoms (“C1-C24 alkyl”). In some embodiments, an alkyl group has 1 to 12 carbon atoms (“C1-C12 alkyl”). In some embodiments, an alkyl group has 1 to 8 carbon atoms (“Ci-Cs alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C1-C6 alkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C1-C5 alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“Ci-C4alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C1-C3 alkyl”). In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C1-C2 alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“Ci alkyl”). In some embodiments, an alkyl group has 2 to 6 carbon atoms (“C2- Cealkyl”). Examples of Ci-C6alkyl groups include methyl (Ci), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), iso-butyl (C4), n-pentyl (C5), 3- pentanyl (Cs), amyl (Cs), neopentyl (Cs), 3-methyl-2-butanyl (Cs), tertiary amyl (Cs), and n- hexyl (Ce). Additional examples of alkyl groups include n-heptyl (C7), n-octyl (Cs) and the like. Each instance of an alkyl group may be independently optionally substituted, i.e., unsubstituted (an“unsubstituted alkyl”) or substituted (a“substituted alkyl”) with one or more substituents; e.g., for instance from 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, the alkyl group is unsubstituted Ci- 10 alkyl (e.g., -CH 3 ). In certain embodiments, the alkyl group is substituted Ci- 6 alkyl.

As used herein,“alkenyl” refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 24 carbon atoms, one or more carbon-carbon double bonds, and no triple bonds (“C 2 -C 24 alkenyl”). In some embodiments, an alkenyl group has 2 to 10 carbon atoms (“C 2 -C 10 alkenyl”). In some embodiments, an alkenyl group has 2 to 8 carbon atoms (“C 2 -C 8 alkenyl”). In some embodiments, an alkenyl group has 2 to 6 carbon atoms (“C 2 -C 6 alkenyl”).

In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C2-C5 alkenyl”). In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C2-C4 alkenyl”). In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C2-C3 alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C2 alkenyl”). The one or more carbon- carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples of C2-C4 alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1- butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. Examples of CYO, alkenyl groups include the aforementioned C2-4 alkenyl groups as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6), and the like. Additional examples of alkenyl include heptenyl (C7), octenyl (Cs), octatrienyl (Cs), and the like. Each instance of an alkenyl group may be independently optionally substituted, i.e., unsubstituted (an“unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) with one or more substituents e.g., for instance from 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, the alkenyl group is unsubstituted C2-10 alkenyl. In certain embodiments, the alkenyl group is substituted C2-6 alkenyl.

As used herein, the term“alkynyl” refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 24 carbon atoms, one or more carbon-carbon triple bonds (“C 2 -C 24 alkenyl”). In some embodiments, an alkynyl group has 2 to 10 carbon atoms (“C 2 -C 10 alkynyl”). In some embodiments, an alkynyl group has 2 to 8 carbon atoms (“C 2 -C 8 alkynyl”).

In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C 2 -C 6 alkynyl”). In some embodiments, an alkynyl group has 2 to 5 carbon atoms (“C 2 -C 5 alkynyl”). In some embodiments, an alkynyl group has 2 to 4 carbon atoms (“C 2 -C 4 alkynyl”). In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C 2 -C 3 alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (“C2 alkynyl”). The one or more carbon- carbon triple bonds can be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl). Examples of C2-C4 alkynyl groups include ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1- butynyl (C4), 2-butynyl (C4), and the like. Each instance of an alkynyl group may be independently optionally substituted, i.e., unsubstituted (an“unsubstituted alkynyl”) or substituted (a“substituted alkynyl”) with one or more substituents e.g., for instance from 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, the alkynyl group is unsubstituted C2- 10 alkynyl. In certain embodiments, the alkynyl group is substituted C2- 6 alkynyl.

As used herein, the term "heteroalkyl," refers to a non-cyclic stable straight or branched chain, or combinations thereof, including at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, Si, and S, and wherein the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) O, N, P, S, and Si may be placed at any position of the heteroalkyl group. Exemplary heteroalkyl groups include, but are not limited to: -CH2-CH2-O-CH3, -CH2-CH2-NH- CH 3 , -CH2-CH2-N(CH 3 )-CH 3 , -CH2-S-CH2-CH3, -CH2-CH2, -NHCH2-, -C(0)NH-, - C(0)N(CH 3 ), -C(0)N(CH 2 CH 3 )-, -C(0)N(CH 2 CF 3 )-, -S(0)-CH 3 , -CH2-CH 2 -S(0)2-CH 3 , - CH=CH-0-CH , -Si(CH )3, -CH2-CH=N-OCH , -CH=CH-N(CH )-CH , -0-CH3, and -O-CH2- CH3. Up to two or three heteroatoms may be consecutive, such as, for example, -CH2-NH- OCH3 and -CH2-0-Si(CH3)3- Where "heteroalkyl" is recited, followed by recitations of specific heteroalkyl groups, such as -CH2O, -NR C R D , or the like, it will be understood that the terms heteroalkyl and -CH2O or -NR C R D are not redundant or mutually exclusive. Rather, the specific heteroalkyl groups are recited to add clarity. Thus, the term "heteroalkyl" should not be interpreted herein as excluding specific heteroalkyl groups, such as -CH2O, -NR C R D , or the like.

As used herein,“aryl” refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 p electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (“C6-C14 aryl”). In some embodiments, an aryl group has six ring carbon atoms (“C 6 aryl”; e.g., phenyl). In some embodiments, an aryl group has ten ring carbon atoms (“Cio aryl”; e.g., naphthyl such as 1 -naphthyl and 2-naphthyl). In some embodiments, an aryl group has fourteen ring carbon atoms (“C14 aryl”; e.g., anthracyl). An aryl group may be described as, e.g., a C 6 -Cio-membered aryl, wherein the term“membered” refers to the non-hydrogen ring atoms within the moiety. Aryl groups include phenyl, naphthyl, indenyl, and tetrahydronaphthyl. Each instance of an aryl group may be independently optionally substituted, i.e., unsubstituted (an “unsubstituted aryl”) or substituted (a“substituted aryl”) with one or more substituents. In certain embodiments, the aryl group is unsubstituted C6-C14 aryl. In certain embodiments, the aryl group is substituted C6-C14 aryl.

As used herein,“heteroaryl” refers to a radical of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 p electrons shared in a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur (“5-10 membered heteroaryl”). In heteroaryl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. Heteroaryl bicyclic ring systems can include one or more heteroatoms in one or both rings.“Heteroaryl” also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused (aryl/heteroaryl) ring system. Bicyclic heteroaryl groups wherein one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and the like) the point of attachment can be on either ring, / ' . e. , either the ring bearing a heteroatom (e.g., 2-indolyl) or the ring that does not contain a heteroatom (e.g., 5-indolyl). A heteroaryl group may be described as, e.g., a 6-10-membered heteroaryl, wherein the term“membered” refers to the non-hydrogen ring atoms within the moiety.

In some embodiments, a heteroaryl group is a 5-10 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In some embodiments, a heteroaryl group is a 5-8 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In some embodiments, a heteroaryl group is a 5-6 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In some embodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Each instance of a heteroaryl group may be independently optionally substituted, i.e., unsubstituted (an“unsubstituted heteroaryl”) or substituted (a “substituted heteroaryl”) with one or more substituents. In certain embodiments, the heteroaryl group is unsubstituted 5-14 membered heteroaryl. In certain embodiments, the heteroaryl group is substituted 5-14 membered heteroaryl.

Exemplary 5-membered heteroaryl groups containing one heteroatom include, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, without limitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, without limitation, triazolyl, oxadiazolyl, and thiadiazolyl.

Exemplary 5-membered heteroaryl groups containing four heteroatoms include, without limitation, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, without limitation, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6- membered heteroaryl groups containing three or four heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, without limitation, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6- bicyclic heteroaryl groups include, without limitation, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, without limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Other exemplary heteroaryl groups include heme and heme derivatives “heteroaryl” also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more heterocycloalkyl groups wherein the point of attachment is on the heteroaryl ring, and in such instances, the number of carbons continue to designate the number of carbons in the heteroaryl ring system. Exemplary ring systems of this type include 7,8-dihydro-5H-pyrano[4,3-b]pyridine and l,4,6,7-tetahydropyrano[4,3-b]pyrrole.

As used herein,“cycloalkyl” refers to a radical of a non-aromatic cyclic hydrocarbon group having from 3 to 10 ring carbon atoms (“C 3 -C 10 cycloalkyl”) and zero heteroatoms in the non-aromatic ring system. In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms (“C 3 -C 8 cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C 3 -C 6 cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C 3 -C 6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C 5 -C 10 cycloalkyl”). A cycloalkyl group may be described as, e.g., a C 4 -C 7 -membered cycloalkyl, wherein the term“membered” refers to the non-hydrogen ring atoms within the moiety. Exemplary C 3 -C 6 cycloalkyl groups include, without limitation, cyclopropyl (C 3 ), cyclopropenyl (C 3 ), cyclobutyl (C 4 ), cyclobutenyl (C 4 ), cyclopentyl (C 5 ), cyclopentenyl (C 5 ), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and the like. Exemplary C3-C8 cycloalkyl groups include, without limitation, the aforementioned C 3 -C 6 cycloalkyl groups as well as cycloheptyl (C 7 ), cycloheptenyl (C 7 ), cycloheptadienyl (C 7 ), cycloheptatrienyl (C 7 ), cyclooctyl (Cs), cyclooctenyl (Cs), cubanyl (Cs), bicyclo[l. l. l]pentanyl (Cs),

bicyclo[2.2.2]octanyl (Cs), bicyclo[2.1.1]hexanyl (Od, bicyclo[3.1.1]heptanyl (C 7 ), and the like. Exemplary C 3 -C 10 cycloalkyl groups include, without limitation, the aforementioned C 3 -C 8 cycloalkyl groups as well as cyclononyl (C 9 ), cyclononenyl (C 9 ), cyclodecyl (C 10 ), cyclodecenyl (C 10 ), octahydro-1 //-indcnyl (C 9 ), decahydronaphthalenyl (C 10 ), spiro[4.5]decanyl (C 10 ), and the like. As the foregoing examples illustrate, in certain embodiments, the cycloalkyl group is either monocyclic (“monocyclic cycloalkyl”) or contain a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic cycloalkyl”) and can be saturated or can be partially unsaturated. “Cycloalkyl” also includes ring systems wherein the cycloalkyl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is on the cycloalkyl ring, and in such instances, the number of carbons continue to designate the number of carbons in the cycloalkyl ring system. Each instance of a cycloalkyl group may be independently optionally substituted, i.e., unsubstituted (an“unsubstituted cycloalkyl”) or substituted (a“substituted cycloalkyl”) with one or more substituents. In certain embodiments, the cycloalkyl group is unsubstituted C 3 -C 10 cycloalkyl. In certain embodiments, the cycloalkyl group is a substituted C3-C 10 cycloalkyl.

“Heterocyclyl” as used herein refers to a radical of a 3- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“3-10 membered heterocyclyl”). In heterocyclyl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. A heterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”) or a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic heterocyclyl”), and can be saturated or can be partially unsaturated. Heterocyclyl bicyclic ring systems can include one or more heteroatoms in one or both rings. “Heterocyclyl” also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more cycloalkyl groups wherein the point of attachment is either on the cycloalkyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl or aryl or heteroaryl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclyl ring system. A heterocyclyl group may be described as, e.g., a 3-7-membered heterocyclyl, wherein the term “membered” refers to the non-hydrogen ring atoms, i.e., carbon, nitrogen, oxygen, sulfur, boron, phosphorus, and silicon, within the moiety. Each instance of heterocyclyl may be independently optionally substituted, i.e., unsubstituted (an“unsubstituted heterocyclyl”) or substituted (a “substituted heterocyclyl”) with one or more substituents. In certain embodiments, the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. In certain embodiments, the heterocyclyl group is substituted 3-10 membered heterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“5-10 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-8 membered non aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”). In some embodiments, the 5-6 membered heterocyclyl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclyl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclyl has one ring heteroatom selected from nitrogen, oxygen, and sulfur.

Exemplary 3-membered heterocyclyl groups containing one heteroatom include, without limitation, azirdinyl, oxiranyl, thiorenyl. Exemplary 4-membered heterocyclyl groups containing one heteroatom include, without limitation, azetidinyl, oxetanyl and thietanyl.

Exemplary 5-membered heterocyclyl groups containing one heteroatom include, without limitation, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groups containing two heteroatoms include, without limitation, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclyl groups containing three heteroatoms include, without limitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containing one heteroatom include, without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6- membered heterocyclyl groups containing two heteroatoms include, without limitation, piperazinyl, morpholinyl, dithianyl, dioxanyl. Exemplary 6-membered heterocyclyl groups containing two heteroatoms include, without limitation, triazinanyl. Exemplary 7-membered heterocyclyl groups containing one heteroatom include, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary 8-membered heterocyclyl groups containing one heteroatom include, without limitation, azocanyl, oxecanyl and thiocanyl. Exemplary 5-membered heterocyclyl groups fused to a O, aryl ring (also referred to herein as a 5,6-bicyclic heterocyclic ring) include, without limitation, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl, and the like. Exemplary 6-membered heterocyclyl groups fused to an aryl ring (also referred to herein as a 6,6-bicyclic heterocyclic ring) include, without limitation, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.

As used herein,“cyano” refers to the radical -CN.

As used herein,“halo” or“halogen,” independently or as part of another substituent, mean, unless otherwise stated, a fluorine (F), chlorine (Cl), bromine (Br), or iodine (I) atom.

As used herein,“haloalkyl” can include alkyl structures that are substituted with one or more halo groups or with combinations thereof. For example, the terms“fluoroalkyl” includes haloalkyl groups in which the halo is fluorine (e.g., -Ci-Ce alkyl-CF3, -C1-C6 alkyl-C2F). Non limiting examples of haloalkyl include trifluoroethyl, trifluoropropyl, trifluoromethyl, fluoromethyl, diflurormethyl, and fluroisopropyl.

As used herein,“hydroxy” refers to the radical -OH.

As used herein,“nitro” refers to -NO2.

As used herein,“oxo” refers to =0, in which both bonds from the oxygen are connected to the same atom. For example, a carbon atom substituted with oxo froms a carbonyl group - C=0.

Two or more substituents may optionally be joined to form aryl, heteroaryl, cycloalkyl, or heterocyclyl groups. Such so-called ring-forming substituents are typically, though not necessarily, found attached to a cyclic base structure. In one embodiment, the ring-forming substituents are attached to adjacent members of the base structure. For example, two ring forming substituents attached to adjacent members of a cyclic base structure create a fused ring structure. In another embodiment, the ring-forming substituents are attached to a single member of the base structure. For example, two ring- forming substituents attached to a single member of a cyclic base structure create a spirocyclic structure. In yet another embodiment, the ring forming substituents are attached to non-adjacent members of the base structure.

Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various isomeric forms, e.g., enantiomers and/or diastereomers. For example, the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses. See, for example, Jacques et al. ,

Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al, Tetrahedron 33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions p. 268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). The invention additionally encompasses compounds described herein as individual isomers substantially free of other isomers, and alternatively, as mixtures of various isomers.

As used herein, a pure enantiomeric compound is substantially free from other enantiomers or stereoisomers of the compound (i.e., in enantiomeric excess). In other words, an “S” form of the compound is substantially free from the“R” form of the compound and is, thus, in enantiomeric excess of the“R” form. The term“enantiomerically pure” or“pure enantiomer” denotes that the compound comprises more than 75% by weight, more than 80% by weight, more than 85% by weight, more than 90% by weight, more than 91% by weight, more than 92% by weight, more than 93% by weight, more than 94% by weight, more than 95% by weight, more than 96% by weight, more than 97% by weight, more than 98% by weight, more than 99% by weight, more than 99.5% by weight, or more than 99.9% by weight, of the enantiomer. In certain embodiments, the weights are based upon total weight of all enantiomers or

stereoisomers of the compound.

In the compositions provided herein, an enantiomerically pure compound can be present with other active or inactive ingredients. For example, a pharmaceutical composition comprising enantiomerically pure R-compound can comprise, for example, about 90% excipient and about 10% enantiomerically pure R-compound. In certain embodiments, the

enantiomerically pure R-compound in such compositions can, for example, comprise, at least about 95% by weight R-compound and at most about 5% by weight S-compound, by total weight of the compound. For example, a pharmaceutical composition comprising

enantiomerically pure S-compound can comprise, for example, about 90% excipient and about 10% enantiomerically pure S-compound. In certain embodiments, the enantiomerically pure S- compound in such compositions can, for example, comprise, at least about 95% by weight S- compound and at most about 5% by weight R-compound, by total weight of the compound. In certain embodiments, the active ingredient can be formulated with little or no excipient or carrier.

Compound described herein may also comprise one or more isotopic substitutions. For example, H may be in any isotopic form, including 1 H, 2 H (D or deuterium), and 3 H (T or tritium); C may be in any isotopic form, including 12 C, 13 C, and 14 C; O may be in any isotopic form, including 16 0 and 18 0; and the like.

Many of the terms given above may be used repeatedly in the definition of a formula or group and in each case have one of the meanings given above, independently of one another.

It will be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.

Contemplated equivalents of the compounds described above include compounds which otherwise correspond thereto, and which have the same general properties thereof (e.g., the ability to inhibit the formation of TDP-43 inclusions), wherein one or more simple variations of substituents are made which do not adversely affect the efficacy of the compound. In general, the compounds of the present invention may be prepared by the methods illustrated in the general reaction schemes as, for example, described below, or by modifications thereof, using readily available starting materials, reagents and conventional synthesis procedures. In these reactions, it is also possible to make use of variants which are in themselves known, but are not mentioned here.

For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover. Also for purposes of this invention, the term "hydrocarbon" is contemplated to include all permissible compounds having at least one hydrogen and one carbon atom. In a broad aspect, the permissible hydrocarbons include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic organic compounds which can be substituted or unsubstituted.

Pharmaceutical Compositions and Routes of Administration

Pharmaceutical compositions containing compounds described herein such as a compound of Formula (I) or (II) or pharmaceutically acceptable salt thereof can be used to treat or ameliorate a disorder described herein, for example, a neurodegenerative disease, a cancer, an ophthalmological disease (e.g., a retinal disease), or a viral infection. The amount and concentration of compounds of Formula (I) or (II) in the pharmaceutical compositions, as well as the quantity of the pharmaceutical composition administered to a subject, can be selected based on clinically relevant factors, such as medically relevant characteristics of the subject (e.g., age, weight, gender, other medical conditions, and the like), the solubility of compounds in the pharmaceutical compositions, the potency and activity of the compounds, and the manner of administration of the pharmaceutical compositions. For further information on Routes of Administration and Dosage Regimes the reader is referred to Chapter 25.3 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.

While it is possible for a compound of the present invention to be administered alone, it is preferable to administer the compound as a pharmaceutical formulation (composition), where the compound is combined with one or more pharmaceutically acceptable diluents, excipients or carriers. The compounds according to the invention may be formulated for administration in any convenient way for use in human or veterinary medicine. In certain embodiments, the compound included in the pharmaceutical preparation may be active itself, or may be a prodrug, e.g., capable of being converted to an active compound in a physiological setting. Regardless of the route of administration selected, the compounds of the present invention, which may be used in a suitable hydrated form, and/or the pharmaceutical compositions of the present invention, are formulated into pharmaceutically acceptable dosage forms such as described below or by other conventional methods known to those of skill in the art.

Thus, another aspect of the present invention provides pharmaceutically acceptable compositions comprising a therapeutically effective amount of one or more of the compounds described above, formulated together with one or more pharmaceutically acceptable carriers (additives) and/or diluents. As described in detail below, the pharmaceutical compositions of the present invention may be specially formulated for administration in solid or liquid form, including those adapted for the following: (1) oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), lozenges, dragees, capsules, pills, tablets (e.g., those targeted for buccal, sublingual, and systemic absorption), boluses, powders, granules, pastes for application to the tongue; (2) parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; (3) topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin; (4) intravaginally or intrarectally, for example, as a pessary, cream or foam; (5) sublingually; (6) ocularly; (7) transdermally; (8) transmucosally; (9) nasally; or (10) intrathecally. Additionally, compounds can be implanted into a patient or injected using a drug delivery system. See, for example, Urquhart, et al., (1994) Ann Rev Pharmacol Toxicol 24:199-236; Lewis, ed.“Controlled Release of Pesticides and Pharmaceuticals” (Plenum Press, New York, 1981); U.S. Patent No. 3,773,919; and U.S. Patent No. 35 3,270,960.

The phrase "therapeutically effective amount" as used herein means that amount of a compound, material, or composition comprising a compound of the present invention which is effective for producing some desired therapeutic effect, e.g., by inhibiting TDP-43 inclusions, in at least a sub-population of cells in an animal and thereby blocking the biological consequences of that function in the treated cells, at a reasonable benefit/risk ratio applicable to any medical treatment.

The phrases "systemic administration," "administered systemically," "peripheral administration" and "administered peripherally" as used herein mean the administration of a compound, drug or other material other than directly into the central nervous system, such that it enters the patient's system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.

The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The phrase "pharmaceutically acceptable carrier" as used herein means a

pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject antagonists from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate;

(13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; (21) cyclodextrins such as Captisol®; and (22) other non-toxic compatible substances employed in pharmaceutical formulations.

The term "pharmaceutically acceptable salt" is meant to include salts of the active compounds that are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,

monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, e.g., Berge et al, Journal of Pharmaceutical Science 66: 1-19 (1977)). Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts. These salts may be prepared by methods known to those skilled in the art. Other pharmaceutically acceptable carriers known to those of skill in the art are suitable for the present invention.

Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha- tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Formulations of the present invention include those suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and/or parenteral administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred per cent, this amount will range from about 1 per cent to about ninety-nine percent of active ingredient, preferably from about 5 per cent to about 70 per cent, most preferably from about 10 per cent to about 30 per cent.

Methods of preparing these formulations or compositions include the step of bringing into association a compound of the present invention with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

Formulations of the invention suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. A compound of the present invention may also be administered as a bolus, electuary or paste.

In solid dosage forms of the invention for oral administration (capsules, tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and (10) coloring agents. In the case of capsules, tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceutical compositions of the present invention, such as dragees, capsules, pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They may be sterilized by, for example, filtration through a bacteria- retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

Liquid dosage forms for oral administration of the compounds of the invention include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

Formulations of the pharmaceutical compositions of the invention for rectal, vaginal, or urethral administration may be presented as a suppository, which may be prepared by mixing one or more compounds of the invention with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.

Alternatively or additionally, compositions can be formulated for delivery via a catheter, stent, wire, or other intraluminal device. Delivery via such devices may be especially useful for delivery to the heart, lung, bladder, urethra, ureter, rectum, or intestine. Furthermore, compositions can be formulated for delivery via a dialysis port.

Ophthalmic formulations, eye ointments, powders, solutions and the like, are also contemplated as being within the scope of this invention.

Exemplary modes of administration include, but are not limited to, injection, infusion, instillation, inhalation, or ingestion.“Injection” includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intraventricular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, sub capsular, subarachnoid, intraspinal, intracerebro spinal, and intrasternal injection and infusion. In some embodiments, the compositions are administered by intravenous infusion or injection.

The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. Pharmaceutical compositions of this invention suitable for parenteral administration comprise one or more compounds of the invention in combination with one or more

pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

Injectable depot forms are made by forming microencapsule matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissue.

When the compounds of the present invention are administered as pharmaceuticals, to humans and animals, they can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.

The addition of the active compound of the invention to animal feed is preferably accomplished by preparing an appropriate feed premix containing the active compound in an effective amount and incorporating the premix into the complete ration. Alternatively, an intermediate concentrate or feed supplement containing the active ingredient can be blended into the feed. The way in which such feed premixes and complete rations can be prepared and administered are described in reference books (such as "Applied Animal Nutrition", W.H.

Freedman and CO., San Francisco, U.S.A., 1969 or "Livestock Feeds and Feeding" O and B books, Corvallis, Ore., U.S.A., 1977).

Methods of introduction may also be provided by rechargeable or biodegradable devices. Various slow release polymeric devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including proteinacious biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non- degradable polymers, can be used to form an implant for the sustained release of a compound at a particular target site.

Preferably, the subject is a mammal. The mammal can be a human, non-human primate, mouse, rat, dog, cat, horse, or cow, but are not limited to these examples. Mammals other than humans can be advantageously used as subjects that represent animal models of disorders associated with neurodegenerative disease or disorder, cancer, or viral infections.

In addition, the methods described herein can be used to treat domesticated animals and/or pets. A subject can be male or female. A subject can be one who has been previously diagnosed with or identified as suffering from or having a neurodegenerative disease or disorder, a disease or disorder associated with cancer, a disease or disorder associated with viral infection, or one or more complications related to such diseases or disorders but need not have already undergone treatment.

Dosages

Actual dosage levels of the active ingredients in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factors including the activity of the particular compound of the present invention employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the invention employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

The compound and the pharmaceutically active agent can be administrated to the subject in the same pharmaceutical composition or in different pharmaceutical compositions (at the same time or at different times). When administrated at different times, the compound and the pharmaceutically active agent can be administered within 5 minutes, 10 minutes, 20 minutes, 60 minutes, 2 hours, 3 hours, 4, hours, 8 hours, 12 hours, 24 hours of administration of the other agent. When the inhibitor and the pharmaceutically active agent are administered in different pharmaceutical compositions, routes of administration can be different.

The amount of compound that can be combined with a carrier material to produce a single dosage form will generally be that amount of the inhibitor that produces a therapeutic effect. Generally out of one hundred percent, this amount will range from about 0.1 % to 99% of inhibitor, preferably from about 5% to about 70%, most preferably from 10% to about 30%.

Toxicity and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED50. Compositions that exhibit large therapeutic indices are preferred.

The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized.

The therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the therapeutic which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Levels in plasma may be measured, for example, by high performance liquid chromatography. The effects of any particular dosage can be monitored by a suitable bioassay.

The dosage may be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment. Generally, the compositions are administered so that the compound of Formula (I) is given at a dose from 1 ng/kg to 200 mg/kg, 10 ng/kg to 100 mg/kg, 10 ng/kg to 50 mg/kg, 100 ng/kg to 20 mg/kg, 100 ng/kg to 10 mg/kg, 100 ng/kg to 1 mg/kg, 1 pg/kg to 100 mg/kg, 1 pg/kg to 50 mg/kg, 1 pg/kg to 20 mg/kg, 1 pg/kg to 10 mg/kg, 1 pg/kg to 1 mg/kg, 10 pg/kg to 10 mg/kg, 10 pg/kg to 50 mg/kg, 10 pg/kg to 20 mg/kg, 10 pg/kg to 10 mg/kg, 10 pg/kg to 1 mg/kg, 100 pg/kg to 50 mg/kg, 100 pg/kg to 20 mg/kg, 1 mg/kg to 100 mg/kg, 1 mg/kg to 50 mg/kg, 1 mg/kg to 20 mg/kg, 1 mg/kg to 10 mg/kg, 1 pg/kg to 10 mg/kg, 10 mg/kg to 100 mg/kg, 10 mg/kg to 50 mg/kg, 10 mg/kg to 20 mg/kg, or 50 mg/kg to 100 mg/kg. It is to be understood that ranges given here include all intermediate ranges, e.g., the range 1 mg/kg to 10 mg/kg includes 1 mg/kg to 2 mg/kg, 1 mg/kg to 3 mg/kg, 1 mg/kg to 4 mg/kg, 1 mg/kg to 5 mg/kg, 1 mg/kg to 6 mg/kg, 1 mg/kg to 7 mg/kg, 1 mg/kg to 8 mg/kg, 1 mg/kg to 9 mg/kg, 2 mg/kg to 10 mg/kg, 3 mg/kg to 10 mg/kg, 4 mg/kg to 10 mg/kg, 5 mg/kg to 10 mg/kg, 6 mg/kg to 10 mg/kg, 7 mg/kg to 10 mg/kg, 8 mg/kg to 10 mg/kg, 9 mg/kg to 10 mg/kg, and the like. It is to be further undertood that the ranges intermediate to the given above are also within the scope of this invention, for example, in the range 1 mg/kg to 10 mg/kg, dose ranges such as 2 mg/kg to 8 mg/kg, 3 mg/kg to 7 mg/kg, 4 mg/kg to 6 mg/kg, and the like.

With respect to duration and frequency of treatment, it is typical for skilled clinicians to monitor subjects in order to determine when the treatment is providing therapeutic benefit, and to determine whether to increase or decrease dosage, increase or decrease administration frequency, discontinue treatment, resume treatment or make other alteration to treatment regimen. The dosing schedule can vary from once a week to daily depending on a number of clinical factors, such as the subject's sensitivity to the drugs. The desired dose can be administered at one time or divided into subdoses, e.g. , 2-4 subdoses and administered over a period of time, e.g., at appropriate intervals through the day or other appropriate schedule. Such sub-doses can be administered as unit dosage forms. In some embodiments, administration is chronic, e.g., one or more doses daily over a period of weeks or months. Examples of dosing schedules are administration daily, twice daily, three times daily or four or more times daily over a period of 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months or more. The present invention contemplates formulation of the subject compounds in any of the aforementioned pharmaceutical compositions and preparations. Furthermore, the present invention contemplates administration via any of the foregoing routes of administration. One of skill in the art can select the appropriate formulation and route of administration based on the condition being treated and the overall health, age, and size of the patient being treated.

EXAMPLES

Examples are provided below to facilitate a more complete understanding of the invention. The following examples illustrate exemplary modes of making and practicing the invention. However, the scope of the invention is not limited to specific embodiments disclosed in these Examples, which are for purposes of illustration only, since alternative methods can be utilized to obtain similar results.

General. All oxygen and/or moisture sensitive reactions were carried out under N2 atmosphere in glassware that was flame-dried under vacuum (0.5 mmHg) and purged with N2 prior to use. All reagents and solvents were purchased from commercial vendors and used as received, or synthesized according to the footnoted references. NMR spectra were recorded on a Bruker 400 (400 MHz 1 H, 75 MHz 13 C) or Varian (400 MHz 1 H, 75 MHz 13 C) spectrometer. Proton and carbon chemical shifts are reported in ppm (d) referenced to the NMR solvent. Data are reported as follows: chemical shifts, multiplicity (br = broad, s = singlet, t = triplet, q = quartet, m = multiplet; coupling constant (s) in Hz). Unless otherwise indicated NMR data were collected at 25 °C. Flash chromatography was performed using 100-200 mesh Silica Gel. Liquid

Chromatography/Mass Spectrometry (LCMS) was performed on Agilent 1200HPLC and 6110MS. Analytical thin layer chromatography (TLC) was performed on 0.2 mm silica gel plates. Visualization was accomplished with UV light and aqueous potassium permanganate (KMnCL) stain followed by heating.

Table 1: Abbreviations

EXAMPLE 1. Synthesis of Compound 171

Scheme: Synthesis of compound 2-chloro-N-(3-chlorophenyl)-5-(N- ethylmethylsulfonamido)benzamide 5 (Compound 171):

Experimental Procedures:

Preparation of compound methyl 5-amino-2-chlorobenzoate (2):

HC1 (gas) was bubbled through a solution of 5-amino-2-chloro-benzoic acid (1 g, 5.8 mmol, 1 eq ) in 25 mL of methanol at -70 °C for about 15 min. and hen the mixture was slowly warmed to 15°C. The mixture was stirred at 15°C for 11 hours 45 mins (15 psi) and then the solution was concentrated in vacuo, diluted with 20 mL of water, and neutralized with 5N NaOH. It was then extracted with 50 mL of ethyl acetate, the organic phase was separated, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 0.8 g of crude compound 2 as a yellow gum, which was used into the next step directly.

Preparation of compound methyl 2-chloro-5-(methylsulfonamido)benzoate (3):

To a solution of compound 2 (0.7 g, 3.8 mmol, 1 eq) in 20 mL of tetrahydrofuran was added TEA (763.3 mg, 7.5 mmol, 2 eq) and MsCl (432.0 mg, 3.8 mmol, 1 eq) at 0°C. The mixture was stirred at 15°C for 0.5 hour. Then the reaction mixture was concentrated under reduced pressure to give a residue, which was purified by column chromatography (Si(¾, eluting with a gradient of petroleum ether : ethyl acetate =10:1 to 1 : 1) to give 150 mg of compound 3 (808.2 pmol, 21.4% yield) as a yellow gum.

Preparation of compound methyl 2-chloro-5-(N-ethylmethylsulfonamido)benzoate (4):

3 4

To a solution of compound 3 (0.25 g, 948.1 pmol, 1 eq) in 1 mL of acetonitrile was added iodoethane (221.8 mg, 1.4 mmol, 1.5 eq), K2CO3 (262.1 mg, 1.9 mmol, 2 eq). The mixture was stirred at 70°C for 12 hours. The reaction mixture was filtered and concentrated under reduced pressure to give 260 mg of crude compound 4 as a yellow gum, which was used into next step directly.

Preparation of compound 2-chloro-N-(3-chlorophenyl)-5-(N- ethylmethylsulfonamido)benzamide Compound 171:

4 5 To a solution of methyl 2-chloro-5-[ethyl(methylsulfonyl)amino]benzoate (0.1 g, 342.8 pmol, 1 eq) and 3-chloroaniline (65.6 mg, 514.1 pmol, 1.5 eq) in 3 mL of toluene was added AlMe 3 (2 M, 342.8 pL, 2 eq) at 0°C. The mixture was stirred at 100°C for 1 hour. Then the reaction mixture was partitioned between 10 mL of 1M HC1 and 20 mL of ethyl acetate. The organic phase was separated, washed with 4 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by was purified by prep-TLC (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =3: 1) to give 91.4 mg of compound 171 (232.1 pmol, 67.7% yield, 98.3% purity) as a light yellow solid.

J H NMR (400 MHz, CHLOROF ORM-7) d ppm 8.06 (br s, 1 H) 7.80 (t, 7=1.96 Hz, 1 H) 7.76 (d, 7=2.57 Hz, 1 H) 7.50 - 7.54 (m, 1 H) 7.43 - 7.50 (m, 2 H) 7.32 (t, 7=8.13 Hz, 1 H) 7.15 - 7.21

(m, 1 H) 3.77 (q, 7=7.09 Hz, 2 H) 2.93 (s, 3 H) 1.18 (t, 7=7.09 Hz, 3 H)

LCMS (ESI+): m/z 387.0 (M+H).

EXAMPLE 2. Synthesis of Compound 101

Scheme: Synthesis of compound 2-chloro-N-(3-chlorophenyl)-5-(N- methylmethylsulfonamido)benzamide 6 (Compound 101):

Experimental Procedures:

Preparation of compound methyl 2-chloro-5-(N-methylmethylsulfonamido)benzoate (5):

To a solution of compound 3 (0.1 g, 379.2 pmol, 1 eq) in 2 mL of DMF was added NaH (22.8 mg, 568.8 pmol, 60% purity, 1.5 eq) at 0°C. The mixture was stirred at 0°C for 0.5 hour. Then Mel (80.7 mg, 568.8 pmol, 1.5 eq) was added to the mixture, and the mixture was stirred at 0°C for another 0.5 hour. After reaction, it was partitioned between 3 mL of saturated NH4CI and 8 mL of ethyl acetate. The organic phase was separated, washed with 4 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 0.1 g of crude compound 5 as a yellow solid, which was used into the next step directly. Preparation of compound 2-chloro-N-(3-chlorophenyl)-5-(N- methylmethylsulfonamido)benzamide 6 (Compound 101):

5 6

To a solution of compound 5 (0.1 g, 360.1 pmol, 1 eq) and 3-chloroaniline (68.9 mg,

540.1 pmol, 1.5 eq) in 3 mL of toluene was added AlMe3 (2 M, 180.0 pL, 1 eq) at 0°C. The mixture was stirred at 100°C for 1 hour. Then the reaction mixture was partitioned between 10 mL of HC1 (1M) and 20 mL of ethyl acetate. The organic phase was separated, washed with 4 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (TFA condition) to give 82.4 mg of

Compound 101 (220.8 pmol, 61.3% yield, 100% purity) as a white solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.77 (s, 1 H) 7.91 (t, 7=1.90 Hz, 1 H) 7.69 (d, 7=2.57 Hz, 1 H) 7.57 - 7.64 (m, 2 H) 7.50 - 7.55 (m, 1 H) 7.40 (t, 7=8.07 Hz, 1 H) 7.20 (dd, 7=7.95, 1.22 Hz, 1 H) 3.27 (s, 3 H) 3.03 (s, 3 H)

LCMS (ESI+) : m/z 373.0 (M+H)

EXAMPLE 3. Synthesis of Compound 100

Synthesis of 2-chloro-N-(3-chlorophenyl)-5-(l,l-dioxido-l,2-thiazinan-2-y l)benzamide 4

(Compound 100)

Experimental Procedures:

The synthetic method of compound 2 can be found in the synthesis of Compound 171.

Preparation of compound methyl 2-chloro-5-(l, l-dioxido-l,2-thiazinan-2-yl)benzoate (3):

To a solution of methyl 5-amino-2-chloro-benzoate (0.4 g, 2.2 mmol, 1 eq) in 5 mL of dichloromethane was added TEA (654.2 mg, 6.5 mmol, 899.9 mE, 3 eq) and 4-chlorobutane-l- sulfonyl chloride (494.14 mg, 2.59 mmol, 1.2 eq). The mixture was stirred at 15°C for 12 hours. The reaction mixture was concentrated under reduced pressure to give a residue, which was dissolved in 5 mL of methanol and triethyl amine (TEA, 436.2 mg, 4.3 mmol, 599.9 pL, 2 eq) was added. The mixture was stirred at 70°C for 3 hours and was then concentrated under reduced pressure to give a residue which was purified by column chromatography (SiCh, Petroleum ether : Ethyl acetate = 20: 1 to 4:1) to give 0.3 g of compound 3 (987.6 pmol, 45.8% yield) as a yellow gum.

Preparation of compound 2-chloro-N-(3-chlorophenyl)-5-(l,l-dioxido-l,2-thiazinan-2- yl)benzamide 4 (Compound 100):

To a solution of methyl 2-chloro-5-(l,l-dioxothiazinan-2-yl)benzoate (0.1 g, 329.2 pmol, 1 eq) and 3-chloroaniline (63.0 mg, 493.8 pmol, 1.5 eq) in 2 mL of toluene was added dropwise AlMe 3 (2 M, 329.2 pL, 2 eq) at 0°C. After addition, the resulting mixture was stirred at 100°C for 1 hour. The reaction mixture was partitioned between 10 mL of 1M HC1 and 20 mL of ethyl acetate. The organic phase was separated, washed with 4 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-HPLC (TLA condition) to give 55.6 mg of 4 (137.0 pmol, 41.6% yield, 98.4% purity) as a yellow solid.

J H NMR (400 MHz, CHLOROL ORM-7) d ppm 7.99 (br s, 1 H) 7.80 (t, 7= 1 .83 Hz, 1 H) 7.68 (d, 7=1.83 Hz, 1 H) 7.39 - 7.49 (m, 3 H) 7.31 (t, 7=8.07 Hz, 1 H) 7.17 (dt, 7=7.95, 0.92 Hz, 1 H) 3.67 - 3.80 (m, 2 H) 3.15 - 3.26 (m, 2 H) 2.27 - 2.41 (m, 2 H) 1.93 (dt, 7=11.43, 5.90 Hz, 2 H)

LCMS (ESI+): m/z 399.0 (M+H). The following compounds were prepared analogously cyclizing the appropriate amino - benzoate with 4-chlorobutane-l-sulfonyl chloride, followed by an ester amide exchange reaction with corresponding amine:

Compound 2-chloro-5-( 1 , 1 -dioxido- 1 ,2-thiazinan-2-yl)-N-(3-methoxyphenyl)benzamide) Compound 102):

J H NMR (400 MHz, CHLOROF ORM-7) d ppm 7.80 (br s, 1 H) 7.61 (d, 7=2.32 Hz, 1 H) 7.31 - 7.40 (m, 3 H) 7.20 - 7.24 (m, 1 H) 7.03 (dd, 7=7.95, 1.10 Hz, 1 H) 6.67 (dd, 7=8.19, 1.96 Hz, 1 H) 3.77 (s, 3 H) 3.66 - 3.70 (m, 2 H) 3.12 - 3.17 (m, 2 H) 2.24 - 2.31 (m, 2 H) 1.86 (dt, 7=11.49, 5.87 Hz, 2 H)

LCMS (ESI+): m/z 395.0 (M+H).

Compound 2-chloro-N-(4-chlorophenyl)-5-(l , 1 -dioxido- 1 ,2-thiazinan-2-yl)benzamide

(Compound 103):

J H NMR (400MHz, CHLOROFORM-d) d ppm 7.93 (br s, 1H) 7.60 (d, J=2.2 Hz, 1H) 7.53 (d, J=8.8 Hz, 2H) 7.38 - 7.31 (m, 2H) 7.27 (d, J=8.8 Hz, 2H) 3.70 - 3.60 (m, 2H) 3.17 - 3.09 (m, 2H) 2.31 - 2.20 (m, 2H) 1.87 - 1.80 (m, 2H)

LCMS (ESI+): m/z 399.0 (M+H).

Compound N-(3-chlorophenyl)-5-(l , 1 -dioxido- 1 ,2-thiazinan-2-yl)-2-methylbenzamide (Compound 172):

J H NMR (400MHz, CHLOROFORM-d) d ppm 7.73 (br d, J=15.8 Hz, 2H) 7.43 - 7.37 (m, 2H) 7.33 - 7.29 (m, 1H) 7.25 - 7.21 (m, 2H) 7.10 (dd, J=0.9, 8.0 Hz, 1H) 3.70 - 3.63 (m, 2H) 3.19 - 3.12 (m, 2H) 2.43 (s, 3H) 2.34 - 2.25 (m, 2H) 1.87 (td, J=5.8, 11.5 Hz, 2H)

LCMS (ESI+): m/z 379.0 (M+H). Compound 3-chloro-N-(3-chlorophenyl)-5-(l , 1 -dioxido- 1 ,2-thiazinan-2-yl)benzamide

(Compound 173):

J H NMR (400 MHz, CHLOROFORM-d) d ppm 7.77 (s, 1 H) 7.68 (d, 7=11.13 Hz, 2 H) 7.61 (s, 1 H) 7.44 - 7.47 (m, 1 H) 7.41 (d, 7=9.05 Hz, 1 H) 7.24 (t, 7=8.13 Hz, 1 H) 7.09 (d, 7=8.07 Hz, 1 H) 3.69 - 3.74 (m, 2 H) 3.15 - 3.20 (m, 2 H) 2.26 - 2.34 (m, 2 H) 1.85 - 1.92 (m, 1 H) 1.89 (dt, /=11.46, 5.82 Hz, 1 H)

LCMS (ESI+): m/z 399.0 (M+H).

Compound 4-chloro-N-(3-chlorophenyl)-3-(l , 1 -dioxido- 1 ,2-thiazinan-2-yl)benzamide

(Compound 174):

J H NMR (400MHz, CHLOROFORM-d) d ppm 7.92 (s, 1H) 7.84 (d J=2.0 Hz, 1H) 7.70 - 7.61 (m, 2H) 7.49 - 7.39 (m, 2H) 7.22 - 7.18 (m, 1H) 7.06 (d, J=8.1 Hz, 1H) 3.82 - 3.40 (m, 2H) 3.21 (br s, 2H) 2.31 (br s, 2H) 1.99 - 1.77 (m, 2H)

LCMS (ESI+): m/z 399.0 (M+H)

EXAMPLE 4. Synthesis of Compound 175

Synthesis of 2-chloro-N-(3-chlorophenyl)-5-(methylsulfonamido)benzamide (4) Compound 175:

Experimental Procedures:

Preparation of compound 2:

A mixture of 5-amino-2-chloro-benzoic acid (4 g, 23.3 mmol, 1 eq) in 100 mL of methanol was cooled to 0°C, SOCI2 (6.9 g, 58.3 mmol, 2.5 eq) was added dropwise. and the mixture was stirred at 70°C for 2 hours under N2 atmosphere. It was then partitioned between 60 mL of saturated aqueous sodium bicarbonate solution and 50 mL of ethyl acetate. The organic phase was separated, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 4 g of crude compound 2 as a purple solid, which was used into the next step directly.

Preparation of compound 3:

To a solution of methyl 5-amino-2-chloro-benzoate (500 mg, 2.7 mmol, 1 eq) in 5 mL of tetrahydrofuran was added TEA (327.1 mg, 3.2 mmol, 1.2 eq) and MsCl (308.6 mg, 2.7 mmol, 1 eq) at 0°C. The mixture was stirred at 18°C for 1 hour and was then partitioned between 10 mL of saturated NH4CI and 40 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =1 : 1) to give 330 mg of compound 3 ( 1.3 mmol, 46.5% yield) as a white solid.

Preparation of compound 2-chloro-N-(3-chlorophenyl)-5-(methylsulfonamido)benzamide (4) Compound 175:

3 4 To a solution of methyl 2-chloro-5-(methanesulfonamido)benzoate (50 mg, 189.6 pmol,

1 eq) in 1 mL of toluene was added 3-chloroaniline (29.0 mg, 227.5 pmol, 1.2 eq) and AlMe3 (2 M, 142.2 pL, 1.5 eq) at 0 °C. The mixture was stirred at 100°C for 2 hours. The reaction mixture was quenched by addition 1 mL of methanol and concentrated under reduced pressure to give a residue which was purified by prep- HPLC (TFA condition) to give 21.5 mg of 4

(Compound 175) (59.9 pmol, 31.6% yield, 100.0% purity) as a white solid.

J H NMR (400MHz, CHLOROFORM-d) d ppm 8.18 (s, 1H) 7.75 (s, 1H) 7.56 (d, J=2.6 Hz, 1H) 7.50 - 7.40 (m, 2H) 7.38 - 7.28 (m, 2H) 7.20 - 7.12 (m, 2H) 3.06 (s, 3H)

LCMS (ESI+): m/z 359.0 (M+H).

EXAMPLE 5. Synthesis of Compound 105

Synthesis of compound 2-chloro-N-(3-chlorophenyl)-5-(l,l-dioxido-l,2,6-thiadiazina n-2- yl)benzamide (7) Compound 105:

Experimental Procedures:

Preparation of compound B: To a solution of N-(oxomethylene)sulfamoyl chloride (1 g, 7.1 mmol, 1 eq) in 15 mL of dichloromethane was added 2-methylpropan-2-ol (628.4 mg, 8.5 mmol, 1.2 eq) at 0°C. The mixture was stirred at 0°C for 2 hours. The reaction mixture was concentrated under reduced pressure to give 1.5 g of crude compound B as a white solid, which was used into next step directly.

Preparation of compound 2:

1 2

A mixture of 5-amino-2-chloro-benzoic acid (4 g, 23.3 mmol, 1 eq) in 100 mL of methanol was cooled to 0°C, SOCI2 (6.9 g, 58.3 mmol, 2.5 eq) was added dropwise. The mixture was stirred at 70°C for 2 hours under N2 atmosphere. Then it was partitioned between 60 mL of saturated aqueous sodium bicarbonate solution and 50 mL of ethyl acetate. The organic phase was separated, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 4 g of crude compound 2 as a purple solid, which was used into the next step directly.

Preparation of compound 3:

2 3

To a solution of compound 2 (100 mg, 538.8 pmol, 1 eq ) in 2 mL of dichloromethane was added TEA (81.8 mg, 808.2 pmol, 1.5 eq) and tert-butyl N-chlorosulfonylcarbamate (139.4 mg, 646.5 pmol, 1.2 eq) at 0°C. The mixture was stirred at 20°C for 2 hours. Then it was partitioned between 3 mL of saturated NH4CI and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product which was purified by prep- TLC(SiC>2, eluting with a gradient of petroleum ether:ethyl acetate =2: 1) to give 180 mg of compound 3 (180 mg, 493.4 pmol, 91.6% yield) as a yellow oil. Preparation of compound 4:

3 4

To a solution of compound 3 (600 mg, 1.6 mmol, 1 eq ) in 10 mL of CH3CN was added CS2CO3 (1.6 g, 4.9 mmol, 3 eq) and 1, 3-dibromopropane (996.2 mg, 4.9 mmol, 3 eq). The mixture was stirred at 80°C for 12 hours and was then partitioned between 10 mL of water and 30 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =10:1 to 5:1) to give compound 4 (500 mg, 1.2 mmol, 75.1% yield) as a white solid.

Preparation of compound 5:

To a solution of compound 4 (150 mg, 370.5 mihoI, 1 eq) in 1 mL of methanol and 0.5 mL of H2O was added NaOH (44.5 mg, 1.1 mmol, 3 eq). The mixture was stirred at 20°C for 2 hours. Then it was partitioned between IN HC1 to turn pH to 6, and extracted with 15 mL of ethyl acetate. The organic phase was separated, washed with 5mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the 100 mg of the crude compound 5 as a white solid, which was used into next step directly.

Preparation of compound 6:

To a solution of compound 5 (100 mg, 255.9 piuol, 1 eq) in 1 mL of DMF was added 3- chloroaniline (35.9 mg, 281.5 mitioI, 1.1 eq) and TEA (38.8 mg, 383.8 mitioI, 1.5 eq). The mixture was cooled to 0°C and then a solution of HATU (116.7 mg, 307.0 mitioI, 1.2 eq) in 0.5 mL of DMF was added slowly and stirred at 20°C for 12 hours. It was then partitioned between 3 mL of saturated NH4CI, extracted three times with 15 mL of ethyl acetate, separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- TLC(SiC>2, eluting with a gradient of petroleum ether:ethyl acetate =2: 1) to give 100 mg of compound 6 (199.8 pmol, 78.1% yield) as a white solid.

Preparation of compound 2-chloro-N-(3-chlorophenyl)-5-(l,l-dioxido-l,2,6-thiadiazina n-2- yl)benzamide (7) Compound 105:

6 7

To a solution of compound 6 (100 mg, 199.8 pmol, 1 eq ) in 1 mL of dichloromethane was added TFA (4.6 g, 40.5 mmol, 3.0 mL). The mixture was stirred at 20°C for 1 hour. Then it was concentrated under reduced pressure to give a residue, which was purified by prep- TLC (SiCh, eluting with a gradient of petroleum ether:ethyl acetate =1 : 1) to give 62.5 mg of 7

(Compound 105) (143.9 pmol, 72.0% yield, 92.2% purity) as a white solid.

J H NMR (400MHz, CHLOROFORM-d) d ppm 7.91 (br s, 1H), 7.71 (s, 1H) 7.63 (d, J=2.1 Hz, 1H) 7.41 - 7.35 (m, 3H) 7.23 (t, J=8.1 Hz, 1H) 7.09 (d, J=8.7 Hz, 1H) 4.29 (br t, J=7.6 Hz, 1H) 3.72 - 3.63 (m, 2H) 3.62 - 3.54 (m, 2H) 1.90 (td, J=5.6, 11.1 Hz, 2H)

LCMS (ESI+): m/z 400.0 (M+H).

EXAMPLE 6. Synthesis of Compound 176

Synthesis of compound 2,4-dichloro-N-(3-chlorophenyl)-5-(l,l-dioxidoisothiazolidin -2- yl)benzamide 5 (Compound 176):

Experimental Procedures:

Preparation of compound 2:

To a solution of HNO3 (3.1 g, 33.3 mmol, 2.2 mL, 68% purity, 22.8 eq ) in H2SO4 (2.8 g, 28.1 mmol, 1.5 mL, 98% purity, 19.2 eq) was added methyl 2,4-dichlorobenzoate (0.3 g, 1.5 mmol, 1 eq) at 0°C. After addition, the mixture was stirred at this temperature for 0.5 hours. The resulting mixture was allowed to warm to 25 °C and stirred for 2.5 hours. The reaction mixture was partitioned between 3 mL of ice-water and 6 mL of ethyl acetate. The organic phase was separated, washed with 4 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give a residue, which was purified by prep-TLC (S1O2, Petroleum ether : Ethyl acetate = 3: 1) to give 0.2 g of compound 2 (799.9 pmol, 54.7% yield) as a yellow solid.

Preparation of compound 3:

A mixture of methyl 2,4-dichloro-5-nitro-benzoate (0.2 g, 799.9 pmol, 1 eq), Fe (223.4 mg, 4.0 mmol, 5 eq) and NH4CI (427.9 mg, 8.0 mmol, 10 eq) in 2 mL of methanol and 0.5 mL of water was degassed and purged with N2 for 3 times, and then the mixture was stirred at 70°C for 2 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give 160 mg of crude compound 3 as a yellow solid, which was used into the next step directly.

Preparation of compound 4:

To a solution of methyl 5-amino-2,4-dichloro-benzoate (160 mg, 727.1 mpioΐ, 1 eq) in 2 mL of dichloromethane was added TEA (220.7 mg, 2.2 mmol, 303.6 pL, 3 eq) and 3- chloropropane-l-sulfonyl chloride (193.1 mg, 1.1 mmol, 132.3 pL, 1.5 eq) at 0°C. The mixture was stirred at 25°C for 12 hours. Then it was concentrated under reduced pressure to give a residue. The residue was dissolved in 2 mL of methanol and TEA (220.7 mg, 2.2 mmol, 303.6 pL, 3 eq) was added. The mixture was stirred at 65°C for 3 hours. Then it was concentrated under reduced pressure to remove methanol. The residue was diluted with 3 mL of ethyl acetate, washed three times with 3 mL of sat. NH4CI, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 230 mg of crude compound 4 as a yellow solid, which was used into the next step directly.

Preparation of compound 2,4-dichloro-N-(3-chlorophenyl)-5-(l, l-dioxidoisothiazolidin-2- yl)benzamide 5 (Compound 176):

To a solution of methyl 2, 4-dichloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoate (100 mg, 308.5 pmol, 1 eq) and 3-chloroaniline (51.2 mg, 401.0 pmol, 1.3 eq) in 1 mL of toluene was added dropwise AlMe3 (2 M, 231.4 pL, 1.5 eq) at 0°C. After addition, the resulting mixture was stirred at 100°C for 3 hours. Then it was concentrated under reduced pressure to remove the toluene, then 2 mL of methanol was added. The mixture was filtered, and the filtrate was purified by prep-HPLC (TLA condition) to give 83 mg of 5 (Compound 176) (197.8 pmol, 64.1% yield, 100% purity) as a yellow solid.

J H NMR (400 MHz, CHLOROF ORM-7) d ppm 8.11 (br s, 1 H) 7.88 (s, 1 H) 7.77 (t, 7=1.90 Hz, 1 H) 7.58 (s, 1 H) 7.47 (dd, 7=8.07, 0.98 Hz, 1 H) 7.31 (t, 7=8.07 Hz, 1 H) 7.15 - 7.20 (m, 1 H) 3.77 (t, 7=6.79 Hz, 2 H) 3.34 (t, 7=7.52 Hz, 2 H) 2.59 (quin, 7=7.15 Hz, 2 H)

LCMS (ESI+): m/z 418.9 (M+H).

EXAMPLE 7. Synthesis of Compound 118

Scheme: Synthesis of 2,3-dichloro-N-(3-chlorophenyl)-5-(l,l-dioxidoisothiazolidin -2- yl)benzamide 6 (Compound 118):

5 6

Experimental Procedures:

Preparation of compound 2:

To a solution of 2,3-dichlorobenzoic acid (2 g, 10.5 mmol, 1 eq) in 20 mL of methanol was added SOCI2 (1.5 g, 12.6 mmol, 911.5 pL, 1.2 eq) at 0°C. The mixture was stirred at 65°C for 2 hr. The mixture was concentrated under reduced pressure to give a redidue. Then it was partitioned between 20 mL of water and 60 mL of ethyl acetate. The organic phase was separated, washed with 60 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 2.2 g of crude compound 2 as a yellow oil.

Preparation of compound 3:

To a solution of compound 2 (2 g, 9.8 mmol, 1 eq ) in 6 mL of H2SO4 was added a solution of 1.5 mL of HNO3 in 2 mL of H2SO4 at -20°C. The mixture was stirred at 10°C for 3 hours. The residue was poured into ice water 40 mL, filtered to give 1 g of crude compound 3 as a white solid.

Preparation of compound 4:

A mixture of compound 3 (300 mg, 1.2 mmol, 1 eq), Le (335.1 mg, 6.00 mmol, 5 eq) and NH4CI (641.8 mg, 12.0 mmol, 10 eq) in 5 mL of methanol and 1 mL of H2O, and then the mixture was stirred at 65°C for 2 hours under N2 atmosphere. The mixture was filtered and the filtrate was concentrated under reduced pressure to give the residue. The reaction mixture was partitioned between 20 mL of water and 45 mL of ethyl acetate. The organic phase was separated, washed with 40 mL of brine, dried with anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by prep-TLC (S1O2, eluting with petroleum ether:Ethyl acetate = 2: 1) to give 250 mg of crude compound 4 as a brown solid.

Preparation of compound 5:

A mixture of compound 4 (250 mg, 1.0 mmol, 1 eq ) and 3-chloropropane-l-sulfonyl chloride (261.5 mg, 1.48 mmol, 179.1 pL, 1.3 eq) in 5 mL of dichloromethane was added TEA (344.9 mg, 3.41 mmol, 474.4 pL, 3 eq) at 0°C, and then the mixture was stirred at 25°C for 3 hours. Then the mixture was concentrated under reduced pressure to give a residue. The residue was dissolved in 5 mL of methanol, then added TEA (344.9 mg, 3.4 mmol, 474.4 pL, 3 eq) at 25°C, the mixture was stirred at 70°C for 10 hours. The mixture was concentrated under reduced pressure to give a rediude, the reaction mixture was partitioned between 5 mL of water and 9 mL of ethyl acetate. The organic phase was separated, washed with 12 mL of brine, dried with anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by prep- TLC (S1O2, eluting with petroleum ether:Ethyl acetate = 1 : 1) to give 180 mg of compound 5 (555.3 pmol, 48.9% yield) as a yellow solid.

Preparation of 2,3-dichloro-N-(3-chlorophenyl)-5-(l,l-dioxidoisothiazolidin -2-yl)benzamide 6

(Compound 118):

5 Compound

118

A mixture of compound 5 (70 mg, 215.9 m ihoI, 1 eq), 3-chloroaniline (30.3 mg, 237.5 m mol, 1.1 eq) in 2 mL of toluene was added AlMe3 (2 M, 162.0 pL, 1.5 eq) at 0°C, and then the mixture was stirred at 100°C for 3 hours under N2 atmosphere. After reaction, it was quenched by addition of 5 mL of methanol, and then concentrated under reduced pressure to give a residue, which was purified by prep- HPLC (neutral condition) to give 32.2 mg of 2,3-dichloro- N-(3-chlorophenyl)-5-(l,l-dioxidoisothiazolidin-2-yl)benzami de 6 (Compound 118) (76.5 pmol, 35.4% yield, 99.7 % purity) as a white solid.

J H NMR (400 MHz, METH AN OL-cA) d ppm 7.84 (t, 7=1.96 Hz, 1 H) 7.48 - 7.61 (m, 2 H) 7.30 - 7.40 (m, 2 H) 7.17 (dd, 7=8.01, 1.04 Hz, 1 H) 3.83 (t, 7=6.54 Hz, 2 H) 3.50 (t, 7=7.40 Hz, 2 H) 2.54 (quin, 7=6.94 Hz, 2 H).

LCMS (ESI+): m/z 419.0 (M+H).

The following compounds were prepared analogously to compound 2,3-dichloro-N-(3- chlorophenyl)-5-(l,l-dioxidoisothiazolidin-2-yl)benzamide 6 using different amines:

N-(3-bromo-5-chlorophenyl)-2,3-dichloro-5-(l,l-dioxidoisothi azolidin-2-yl)benzamide Compound 143:

1H NMR (400MHz, CHLOROFORM-d) d ppm 7.82 (br s, 1H) 7.67 (s, 1H) 7.60 (s, 1H) 7.47 (d, J=2.8 Hz, 1H) 7.32 (d, J=2.8 Hz, 1H) 7.27 (t, J=1.7 Hz, 1H) 3.72 (t, J=6.5 Hz, 2H) 3.36 (t, J=7.5 Hz, 2H) 2.52 (quin, J=7.0 Hz, 2H)

LCMS (ESI+): m/z 499.0 (M+H). 2,3-dichloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(3-(triflu oromethyl)phenyl)benzamide Compound 177:

J H NMR (400MHz, DMSO-d6) d ppm 10.99 (s, 1H) 8.19 (s, 1H) 7.89 (br d, J=8.1 Hz, 1H) 7.63 (t, J=8.0 Hz, 1H) 7.55 (d, J=2.7 Hz, 1H) 7.51 (d, J=7.8 Hz, 1H) 7.37 (d, J=2.8 Hz, 1H) 3.82 (t, J=6.5 Hz, 2H) 3.61 (t, J=7.3 Hz, 2H) 2.48 - 2.40 (m, 2H)

LCMS (ESI+): m/z 453.0 (M+H).

2,3-dichloro-N-(3,5-difluorophenyl)-5-(l,l-dioxidoisothia zolidin-2-yl)benzamide

Compound 168:

J H NMR (400 MHz, DMSO-d6) d ppm 2.40 - 2.46 (m, 2 H) 3.61 (t, J = 7.34 Hz, 2 H) 3.81 (t, J = 6.48 Hz, 2 H) 7.02 (tt, / = 9.35, 2.32 Hz, 1 H) 7.35 (d, / = 2.69 Hz, 1 H) 7.41 (dd, / = 9.29, 2.08 Hz, 2 H) 7.55 (d, / = 2.69 Hz, 1 H) 11.04 (s, 1 H).

LCMS (ESI+): m/z 421.0 (M+H).

2,3-dichloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(3-fluo ro-5-methylphenyl)benzamide Compound 169:

1H NMR (400 MHz, DMSO-d6) d ppm 2.31 (s, 3 H) 2.43 (t, J = 7.03 Hz, 2 H) 3.60 (t, J = 7.34 Hz, 2 H) 3.81 (t, / = 6.48 Hz, 2 H) 6.81 (br d, / = 9.41 Hz, 1 H) 7.27 (s, 1 H) 7.31 (d, / = 2.69 Hz, 1 H) 7.43 (br d, J = 11.13 Hz, 1 H) 7.53 (d, / = 2.69 Hz, 1H) 10.76 (s, 1 H)

LCMS (ESI+): m/z 417.0 (M+H). 2,3-dichloro-N-(3-chloro-5-fluorophenyl)-5-(l,l-dioxidoisoth iazolidin-2-yl)benzamide Compound 161:

J H NMR (400 MHz, DMSO-d6) d ppm 11.02 (s, 1 H) 7.63 (s, 1 H) 7.53 - 7.58 (m, 2 H) 7.36 (d, J=2.69 Hz, 1 H) 7.22 (dt, J=8.68, 2.08 Hz, 1 H) 3.82 (t, J=6.54 Hz, 2 H) 3.61 (t, J=7.40 Hz, 2 H) 2.40 - 2.46 (m, 2 H)

LCMS (ESI+): m/z 436.9 (M+H).

N-(3,5-bis(trifluoromethyl)phenyl)-2,3-dichloro-5-(l,l-di oxidoisothiazolidin-2- yl)benzamide Compound 160:

J H NMR (400 MHz, DMSO-76) d ppm 11.33 (s, 1 H) 8.36 (s, 2 H) 7.88 (s, 1 H) 7.58 (d, 7=2.69 Hz, 1 H) 7.43 (d, 7=2.69 Hz, 1 H) 3.82 (t, 7=6.54 Hz, 2 H) 3.62 (t, 7=7.34 Hz, 2 H) 2.44 (t, 7=6.97 Hz, 2 H)

LCMS (ESI+): m/z 521.0 (M+H).

2,3-dichloro-N-(3,5-dichlorophenyl)-5-(l,l-dioxidoisothia zolidin-2-yl)benzamide

Compound 133:

J H NMR (400 MHz, DMSO-de) d ppm 10.99 (s, 1 H) 7.75 (d, 7=1.96 Hz, 2 H) 7.55 (d, 7=2.69 Hz, 1 H) 7.38 (t, 7=1.90 Hz, 1 H) 7.35 (d, 7=2.69 Hz, 1 H) 3.81 (t, 7=6.48 Hz, 2 H) 3.60 (t, 7=7.34 Hz, 2 H) 2.39 - 2.46 (m, 2 H). LCMS (ESI+): m/z 454.9 (M+H).

2,3-dichloro-N-(3-chloro-5-methylphenyl)-5-(l,l-dioxidois othiazolidin-2-yl)benzamide Compound 132:

J H NMR (400 MHz, DMSO -d 6 ) d ppm 10.74 (s, 1 H) 7.65 (s, 1 H) 7.53 (d, 7=2.69 Hz, 1 H) 7.41 (s, 1 H) 7.32 (d, 7=2.69 Hz, 1 H) 7.04 (s, 1 H) 3.81 (t, 7=6.54 Hz, 2 H) 3.60 (t, 7=7.34 Hz, 2 H) 2.43 (quin, 7=6.91 Hz, 2 H) 2.30 (s, 3 H).

LCMS (ESI+): m/z 432.9 (M+H).

2,3-dichloro-N-(3-chloro-5-(trifluoromethyl)phenyl)-5-(l, l-dioxidoisothiazolidin-2- yl)benzamide Compound 164:

J H NMR (400 MHz, DMSO-de) d ppm 11.16 (s, 1 H) 8.05 (br d, 7=1.83 Hz, 2 H) 7.64 (s, 1 H) 7.56 (d, 7=2.69 Hz, 1 H) 7.39 (d, 7=2.69 Hz, 1 H) 3.82 (t, 7=6.48 Hz, 2 H) 3.61 (t, 7=7.34 Hz, 2 H) 2.40 - 2.47 (m, 2 H)

LCMS (ESI+): m/z 486.9 (M+H).

2,3-dichloro-N-(3,5-dimethylphenyl)-5-(l,l-dioxidoisothia zolidin-2-yl)benzamide

Compound 147:

J H NMR (400MHz, DMSO-de) d ppm 10.48 (s, 1 H), 7.50 (d, 7=2.6 Hz, 1 H), 7.34 - 7.23 (m, 3 H), 6.77 (s, 1 H), 3.81 (t, 7=6.4 Hz, 2 H), 3.60 (t, 7=7.2 Hz, 2 H), 2.42 (m, 2 H), 2.26 (s, 6 H).

LCMS (ESI+): m/z 413.0 (M+H).

2,3-dichloro-N-(3-chloro-5-cyanophenyl)-5-(l,l-dioxidoiso thiazolidin-2-yl)benzamide Compound 148:

J H NMR (400MHz, DMSO -d 6 ) d ppm 11.17 (s, 1 H), 8.08 (t, 7=2.0 Hz, 1 H), 8.02 (s, 1 H), 7.83 (t, 7=1.7 Hz, 1 H), 7.56 (d, 7=2.6 Hz, 1 H), 7.38 (d, 7=2.6 Hz, 1 H), 3.81 (t, 7=6.4 Hz, 2 H), 3.61 (t, 7=7.4 Hz, 2 H), 2.47 - 2.40 (m, 2 H).

LCMS (ESI+): m/z 444.0 (M+H).

EXAMPLE 8. Synthesis of Compound 140

The amines of following compounds were made through a Suzuki reaction.

Scheme:

Preparation of compound 2:

To a solution of 3-bromo-5-chloro-aniline (150 mg, 726.5 pmol, 1 eq ), 4-pyridylboronic acid (107.2 mg, 871.9 pmol, 1.2 eq) and K2CO3 (200.9 mg, 1.5 mmol, 2 eq) in 2 mL of dioxane and 0.2 mL of H2O was added Pd(dppf)Cl2 (53.2 mg, 72.7 pmol, 0.1 eq) and purged with N2 for 3 times. The mixture was stirred at 100°C for 12 hour. The reaction mixture was partitioned between 15 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- TLC (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =1 : 1) to give 79 mg of compound 2 (386.0 pmol, 53.1% yield) as a yellow solid.

Then compound 2 was used into an ester amide exchange reaction by using a normal method to give Compound 140.

2,3-dichloro-N-(3-chloro-5-(pyridin-4-yl)phenyl)-5-(l,l-diox idoisothiazolidin-2- yl)benzamide Compound 140:

1H NMR (400 MHz, DMSO-d6) d ppm 10.96 - 11.05 (m, 1 H) 8.67 - 8.74 (m, 2 H) 7.98 - 8.04 (m, 1 H) 7.90 - 7.96 (m, 1 H) 7.73 (br d, J=5.75 Hz, 2 H) 7.67 (s, 1 H) 7.55 (d, J=2.57 Hz, 1 H) 7.34 - 7.40 (m, 1 H) 3.82 (br t, J=6.42 Hz, 2 H) 3.61 (t, J=7.27 Hz, 2 H) 2.38 - 2.45 (m, 2 H)

LCMS (ESI+): m/z 495.9 (M+H).

The following amines were made analogously to Compound 140 using different borate:

2,3-dichloro-N-(3-chloro-5-(pyridin-3-yl)phenyl)-5-(l,l-d ioxidoisothiazolidin-2- yl)benzamide Compound 145:

J H NMR (400 MHz, METHANOL-cA) d ppm 9.05 (d, 7=1.47 Hz, 1 H) 8.76 (d, 7=4.89 Hz, 1 H) 8.60 (dd, 7=8.07, 1.47 Hz, 1 H) 8.04 (t, 7=1.65 Hz, 1 H) 7.94 (dd, 7=8.13, 5.44 Hz, 1 H) 7.86 (t, 7=1.83 Hz, 1 H) 7.61 (t, 7=1.71 Hz, 1 H) 7.55 (d, 7=2.69 Hz, 1 H) 7.40 (d, 7=2.69 Hz, 1 H) 3.83 (t, 7=6.54 Hz, 2 H) 3.50 (t, 7=7.40 Hz, 2 H) 2.51 - 2.58 (m, 2 H)

LCMS (ESI+): m/z 495.9 (M+H). 2,3-dichloro-N-(3-chloro-5-(l-methyl-lH-pyrazol-4-yl)phenyl) -5-(l,l-dioxidoisothiazolidin- 2-yl)benzamide Compound 154:

1H NMR (400 MHz, DMSO-d6) d ppm 10.81 - 10.85 (m, 1 H) 8.16 - 8.20 (m, 1 H) 7.83 - 7.87 (m, 1 H) 7.75 - 7.79 (m, 1 H) 7.62 - 7.68 (m, 1 H) 7.53 - 7.56 (m, 1 H) 7.41 - 7.46 (m, 1 H) 7.34 - 7.37 (m, 1 H) 3.88 (s, 3 H) 3.78 - 3.85 (m, 2 H) 3.55 - 3.65 (m, 2 H) 2.40 - 2.47 (m, 2 H)

LCMS (ESI+): m/z 499.0 (M+H).

2,3-dichloro-N-(5-chloro-[l,r-biphenyl]-3-yl)-5-(l,l-diox idoisothiazolidin-2-yl)benzamide

Compound 146:

J H NMR (400MHz, DMSO -d 6 ) d ppm 10.93 (s, 1 H), 7.87 (d, 7=14.0 Hz, 2 H), 7.65 (d, 7=7.2 Hz, 2 H), 7.57 - 7.35 (m, 6 H), 3.82 (t, 7=6.4 Hz, 2 H), 3.61 (t, 7=7.2 Hz, 2 H), 2.47 - 2.40 (m, 2 H).

LCMS (ESI+): m/z 495.0 (M+H).

EXAMPLE 9. Preparation of 2,3-dichloro-N-(3-chloro-5-isopropylphenyl)-5-(l,l- dioxidoisothiazolidin-2-yl)benzamide Compound 144

Step 1: Preparation of Compound 2

2 To a solution of 3-bromo-5-chloro-aniline (0.2 g, 968.7 pmol, 1 eq), 2-isopropenyl- 4,4,5,5-tetramethyl-l,3,2-dioxaborolane (195.3 mg, 1.2 mmol, 1.2 eq) and K2CO3 (267.8 mg, 1.9 mmol, 2 eq) in 2 mL of dioxane and 0.2 mL of H2O was added Pd(dppf)Ch (70.9 mg, 96.9 pmol, 0.1 eq) and purged with N2 for 3 times. The mixture was stirred at 100°C for 12 hours. The reaction mixture was partitioned between 3 mL of water and 5 mL of ethyl acetate. The organic phase was separated, washed twice with 2 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give a residue, which was purified by prep-TLC (S1O2, eluting with Petroleum ether : Ethyl acetate = 5: 1) to give 83 mg of compound 2 (495.1 pmol, 51.1 % yield) as a green oil.

Step 2: Preparation of compound 3:

2 3

To a solution of 3-chloro-5-isopropenyl-aniline (40 mg, 238.6 pmol, 1 eq) in 1 mL of ethyl acetate was added Pd/C (30 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with ¾ several times. The mixture was stirred under ¾ (15 psi) at 25 °C for 1 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give 42 mg of crude compound 3 as a yellow oil, which was used into the next step directly.

The final compound was obtained by using an ester/amide exchange reaction as used before.

2,3-dichloro-N-(3-chloro-5-isopropylphenyl)-5-(l,l-dioxid oisothiazolidin-2-yl)benzamide Compound 144:

J H NMR (400 MHz, CHLOROFORM-d) d ppm 7.91 (br s, 1 H) 7.51 (s, 1 H) 7.43 (d, J=2.57 Hz, 1 H) 7.23 (d, J=1.71 Hz, 2 H) 6.96 (s, 1 H) 3.66 (t, J=6.48 Hz, 2 H) 3.32 (t, J=7.46 Hz, 2 H) 2.81 (dt, J=13.82, 6.91 Hz, 1 H) 2.48 (quin, J=6.97 Hz, 2 H) 1.17 (d, J=6.85 Hz, 5 H) 1.16 - 1.19 (m, 1 H)

LCMS (ESI+): m/z 461.0 (M+H). 2,3-dichloro-N-(3-chloro-5-(prop-l-en-2-yl)phenyl)-5-(l,l-di oxidoisothiazolidin-2- yl)benzamide Compound 152

Using aniline 2 from above gives the final product.

J H NMR (400 MHz, CHLOROFORM-d) d ppm 7.78 - 7.90 (m, 1 H) 7.61 - 7.67 (m, 1 H) 7.45 - 7.51 (m, 1 H) 7.42 - 7.44 (m, 1 H) 7.25 - 7.36 (m, 1 H) 5.29 - 5.36 (m, 1 H) 5.07 - 5.11 (m, 1 H) 3.65 - 3.75 (m, 2 H) 3.30 - 3.39 (m, 2 H) 2.51 (quin, J=7.00 Hz, 2 H) 2.03 - 2.13 (m, 3 H)

LCMS (ESI+): m/z 459.0 (M+H).

The following compounds were prepared in the same manner of making the aniline and coupling it to the ester.

2,3-dichloro-N-(3-chloro-5-(2-methylprop-l-en-l-yl)phenyl )-5-(l,l-dioxidoisothiazolidin-2- yl)benzamide Compound 151:

1H NMR (400 MHz, CHLOROFORM-d) d ppm 7.89 - 7.98 (m, 1 H) 7.48 - 7.54 (m, 1 H) 7.40 - 7.46 (m, 1 H) 7.19 - 7.23 (m, 1 H) 6.93 - 6.97 (m, 1 H) 6.08 - 6.13 (m, 1 H) 3.62 - 3.72 (m, 2 H) 3.27 - 3.37 (m, 2 H) 2.41 - 2.54 (m, 2 H) 1.76 - 1.87 (m, 6 H)

LCMS (ESI+): m/z 473.0 (M+H). 2,3-dichloro-N-(3-chloro-5-isobutylphenyl)-5-(l,l-dioxidoiso thiazolidin-2-yl)benzamide Compound 159:

1H NMR (400 MHz, CHLOROFORM-d) d ppm 7.88 - 7.95 (m, 1 H) 7.48 - 7.55 (m, 1 H) 7.39 - 7.46 (m, 1 H) 7.21 - 7.28 (m, 1 H) 6.85 - 6.92 (m, 1 H) 3.67 (t, J=6.48 Hz, 2 H) 3.27 - 3.38 (m, 2

H) 2.43 - 2.54 (m, 2 H) 2.38 (d, J=7.21 Hz, 2 H) 1.74 - 1.86 (m, 1 H) 0.84 (d, J=6.60 Hz, 6 H)

LCMS (ESI+): m/z 475.0 (M+H).

(E)-2,3-dichloro-N-(3-chloro-5-(prop-l-en-l-yl)phenyl)-5- (l,l-dioxidoisothiazolidin-2- yl)benzamide Compound 162:

J H NMR (400MHz, DMSO -de) d ppm 10.79 (s, 1 H), 7.67 (t, 7=1.7 Hz, 1 H), 7.56 (s, 1 H), 7.53 (d, 7=2.7 Hz, 1 H), 7.33 (d, 7=2.7 Hz, 1 H), 7.24 (s, 1 H), 6.43 - 6.28 (m, 2 H), 3.81 (t, 7=6.4 Hz, 2 H), 3.60 (t, 7=7.3 Hz, 2 H), 2.43 (quin, 7=6.9 Hz, 2 H), 1.86 (d, 7=5.3 Hz, 3 H).

LCMS (ESI+): m/z 459.0 (M+H).

2,3-dichloro-N-(3-chloro-5-propylphenyl)-5-(l,l-dioxidois othiazolidin-2-yl)benzamide Compound 163:

J H NMR (400MHz, DMSO-de) d ppm 10.76 (s, 1 H), 7.67 (s, 1 H), 7.52 (d, 7=2.6 Hz, 1 H), 7.42 (s, 1 H), 7.32 (d, 7=2.7 Hz, 1 H), 7.05 (s, 1 H), 3.81 (t, 7=6.5 Hz, 2 H), 3.60 (t, 7=7.3 Hz, 2 H), 2.58 - 2.52 (m, 2 H), 2.43 (quin, 7=6.9 Hz, 2 H), 1.58 (sxt, 7=7.4 Hz, 2 H), 0.89 (t, 7=7.3 Hz, 3 H).

LCMS (ESI+): m/z 461.0 (M+H).

EXAMPLE 10. Preparation of 2,3-dichloro-N-(3-chloro-5-(pyridin-2-yl)phenyl)-5-(l,l- dioxidoisothiazolidin-2-yl)benzamide Compound 153

Step 1: Preparation of compound 2

A mixture of 3-bromo-5-chloro-aniline (200 mg, 968.7 mpioΐ, 1 eq), tributyl(2- pyridyl)stannane (427.9 mg, 1.2 mmol, 1.2 eq), K2CO3 (267.8 mg, 1.9 mmol, 2 eq) in 2 mL of dioxane and 0.2 mL of H2O was added Pd(dppf)Cl2 (70.9 mg, 96.9 pmol, 0.1 eq) and purged with N2 for 3 times, and then the mixture was stirred at HO C for 12 hours under N2 atmosphere. Then it was partitioned between 5 mL of water and 10 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give crude product, which was purified by prep-TLC (SiCh, eluting with Petroleum ether : Ethyl acetate= 5: 1) to give 100 mg of compound 2 (488.6 pmol, 50.4% yield) as an orange oil.

Compound 2 was used into an ester amide exchange reaction through a normal method to give

Compound 153.

2,3-dichloro-N-(3-chloro-5-(pyridin-2-yl)phenyl)-5-(l,l-diox idoisothiazolidin-2- yl)benzamide Compound 153:

1H NMR (400 MHz, CHLOROFORM-d) d ppm 8.59 - 8.64 (m, 1 H) 8.33 - 8.39 (m, 1 H) 7.96 - 8.03 (m, 2 H) 7.76 - 7.83 (m, 1 H) 7.65 - 7.73 (m, 2 H) 7.48 (d, J=2.69 Hz, 1 H) 7.26 - 7.32 (m, 1 H) 3.66 - 3.73 (m, 2 H) 3.33 - 3.40 (m, 2 H) 2.47 - 2.56 (m, 2 H)

LCMS (ESI+): m/z 496.0 (M+H).

2,3-dichloro-N-(3-chloro-5-vinylphenyl)-5-(l,l-dioxidoisothi azolidin-2-yl)benzamide Compound 157 was prepared in a similar manner

J H NMR (400MHz, DMSO-d6) d ppm 10.85 (s, 1H) 7.78 - 7.63 (m, 2H) 7.54 (d, J=2.7 Hz, 1H) 7.41 - 7.32 (m, 2H) 6.73 (dd, J=10.9, 17.6 Hz, 1H) 5.88 (d, J=17.6 Hz, 1H) 5.38 (d, J=10.9 Hz, 1H) 3.82 (t, J=6.5 Hz, 2H) 3.64 - 3.58 (m, 2H) 2.44 (br t, J=7.0 Hz, 2H)

LCMS (ESI+): m/z 445.0 (M+H).

EXAMPLE 11. Synthesis of Compound 158

Preparation of 2,3-dichloro-N-(3-chloro-5-ethylphenyl)-5-(l,l-dioxidoisothi azolidin-2- yl)benzamide 4:

Step 1 : Preparation of compound 3:

2 3

A mixture of 3-chloro-5-vinyl-aniline (50 mg, 325.5 pmol, 1 eq), Pd/C (20 mg, 10% purity) in 2 mL of ethyl acetate was degassed and purged with ¾ for three times, and then the mixture was stirred at 25 °C for 10 min under ¾ atmosphere (15 psi). The reaction mixture was filtered and concentrated under reduced pressure to give 50 mg of crude compound 3 as a yellow oil, which was used into next step directly.

Preparation of compound B:

A B

To a solution of compound A (200 mg, 616.9 mihoI, 1 eq ) in 0.3 mL of H2O and 2 mL of methanol was added NaOH (74.0 mg, 1.9 mmol, 3 eq). The mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated under reduced pressure to remove methanol, the crude product was partitioned between 10 mL of HC1 (1M) and 30 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 180 mg of the crude compound B as a yellow solid, which was used into next step directly.

Preparation of 2,3-dichloro-N-(3-chloro-5-ethylphenyl)-5-(l,l-dioxidoisothi azolidin-2- yl)benzamide Compound 158:

To a solution of compound B (50 mg, 161.2 pmol, 1 eq) in 1 mL of DMF was added TEA (19.6 mg, 193.5 pmol, 1.2 eq) and 3-chloro-5-ethyl-aniline (22.6 mg, 145.1 pmol, 0.9 eq). The mixture was cooled to 0°C, HATU (73.6 mg, 193.5 pmol, 1.2 eq) was added and stirred at 25°C for 12 hours. The reaction mixture was filtered, the filtrate was purified by prep- HPLC (TFA condition) to give 18.8 mg of compound 158 (26.0% yield, 100.0% purity) as a white solid

J H NMR (400MHz, DMSO-d6) d ppm 10.75 (s, 1H) 7.67 (s, 1H) 7.52 (d, J=2.7 Hz, 1H) 7.43 (s, 1H) 7.32 (d, J=2.7 Hz, 1H) 7.06 (s, 1H) 3.81 (t, J=6.5 Hz, 2H) 3.60 (t, J=7.3 Hz, 2H) 2.60 (q, J=7.5 Hz, 2H) 2.42 (quin, J=6.9 Hz, 2H) 1.17 (t, J=7.6 Hz, 3H)

LCMS (ESI+): m/z 447.0 (M+H).

EXAMPLE 12. Synthesis of Compound 155

The amines of following compounds were made through a reduction reaction from appropriate nitro compound using the general reaction scheme below.

Preparation of 3-amino-5-methylbenzonitrile:

2

To a solution of 3-methyl-5-nitro-benzonitrile (100 mg, 616.7 m ihoI, 1 eq) in 2 mL of ethanol and 0.5 mL of H2O was added Fe (172.2 mg, 3.1 mmol, 5 eq) and NH4CI (329.9 mg, 6.2 mmol, 10 eq). The mixture was stirred at 80°C for 1 hour. After reaction, the mixture was filtered and the filtrate was partitioned between 5 mL of water and 15 mL of dichloromethane. The organic phase was separated, washed with 5mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 77 mg of crude compound 2 as a yellow oil.

Compound 2 was used in an ester amide exchange reaction by using a normal method to give 2,3-dichloro-N-(3-cyano-5-methylphenyl)-5-(l,l-dioxidoisothi azolidin-2-yl)benzamide Compound 155:

1H NMR (400 MHz, CHLOROFORM-d) d ppm 8.01 (s, 1 H) 7.82 - 7.85 (m, 1 H) 7.67 (s, 1 H) 7.53 (d, J=2.69 Hz, 1 H) 7.41 (d, J=2.69 Hz, 1 H) 7.29 (s, 1 H) 3.80 (t, J=6.54 Hz, 2 H) 3.44 (t, J=7.46 Hz, 2 H) 2.55 - 2.64 (m, 2 H) 2.43 (s, 3 H)

LCMS (ESI+): m/z 424.0 (M+H).

2,3-dichloro-N-(3-chloro-5-methoxyphenyl)-5-(l,l-dioxidoi sothiazolidin-2-yl)benzamide Compound 156 was prepared analogously to using the appropriate nitro compound:

2,3-dichloro-N-(3-chloro-5-methoxyphenyl)-5-(l,l-dioxidoisot hiazolidin-2-yl)benzamide Compound 156:

1H NMR (400 MHz, CHLOROFORM-d) d ppm 7.85 - 7.94 (m, 1 H) 7.53 (d, J=2.69 Hz, 1 H) 7.35 (d, J=2.69 Hz, 1 H) 7.23 (d, J=1.71 Hz, 1 H) 7.19 - 7.22 (m, 1 H) 6.74 (t, J=2.02 Hz, 1 H) 3.81 - 3.87 (m, 3 H) 3.77 (t, J=6.54 Hz, 2 H) 3.42 (t, J=7.52 Hz, 2 H) 2.53 - 2.63 (m, 2 H)

LCMS (ESI+): m/z 449.0 (M+H).

EXAMPLE 13. Synthesis of Compound 178

Scheme: Synthesis of compound 2-chloro-N-(3,5-dichlorophenyl)-3-(l,l-dioxidoisothiazolidin - 2-yl)benzamide 5 (Compound 178):

Experimental Procedures:

Preparation of compound 2:

To a solution of 2-chloro-3-nitro-benzoic acid (1 g, 5.0 mmol, 1 eq) in 15 mL of methanol was added SOCh (708.3 mg, 6.0 mmol, 431.9 pL, 1.2 eq) at 0°C. The mixture was stirred at 65°C for 2 hours. The mixture was concentrated under reduced pressure to give a residue. To the residue was added 15 mL of water, basified by saturated NaHCCb solution until pH~8 at 0°C, extracted three times with 60 mL of ethyl acetate. The combined organic layers were washed twice with 60 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 1.1 g crude compound 2 as a yellow solid.

Preparation of compound 3:

A mixture of compound 2 (1.1 g, 5.10 mmol, 1 eq), Fe (1.4 g, 25.5 mmol, 5 eq) and NH4CI (2.7 g, 51.0 mmol, 10 eq) in 10 mL of methanol and 2 mL of H2O, the mixture was stirred at 60°C for 1 hr under N2 atmosphere. Then it was filtered and the filtrate was concentrated under reduced pressure to give a residue. The reaction mixture was partitioned between 30 mL of water and 60 mL of ethyl acetate. The organic phase was separated, washed with 60 mL of brine, dried with anhydrous Na2SC>4, filtered and concentrated in vacuum to give 950 mg of crude compound 3 as a brown oil.

Preparation of compound 4:

3 4

A mixture of compound 3 (950 mg, 5.1 mmol, 1 eq ) and 3-chloropropane-l-sulfonyl chloride (906.2 mg, 5.12 mmol, 620.7 pL, 1 eq) in 10 mL of dichloromethane was added TEA (1.6 g, 15.3 mmol, 2.1 mL, 3 eq) at 0°C, and then the mixture was stirred at 30°C for 3 hr. It was then concentrated under reduced pressure to give a residue which was dissolved in 10 mL of methanol TEA (1.6 g, 15.3 mmol, 2.14 mL, 3 eq added) was added at 30°C and the mixture was stirred at 70°C for 10 hours. The mixture was concentrated under reduced pressure to give a residue which was partitioned between 20 mL of sat. aqueous NH4CI solution and 45 mL of ethyl acetate. The organic phase was separated, washed twice with 60 mL of brine, dried with anhydrous Na2SC>4, filtered and concentrated in vacuum to give 1.3 g of compound 4 as a brown solid.

Preparation of compound 2-chloro-N-(3,5-dichlorophenyl)-3-(l,l-dioxidoisothiazolidin -2- yl)benzamide 5 (Compound 178):

To a mixture of compound 4 (100 mg, 345.1 pmol, 1 eq), 3,5-dichloroaniline (61.5 mg, 379.7 pmol, 1.1 eq) in 2 mL of toluene was added A1(C¾) 3 (2 M, 258.9 pL, 1.5 eq) at 0°C, and the mixture was stirred at 100°C for 3 hr under N2 atmosphere. The mixture was quenched by addition of 5 mL of methanol and then the mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep- HPLC (TLA condition) to give 28.1 mg of 2- chloro-N-(3,5-dichlorophenyl)-3-(l,l-dioxidoisothiazolidin-2 -yl)benzamide 5 (178) (67.0 pmol, 19.4% yield, 100% purity) as a white solid.

J H NMR (400 MHz, CHLOROF ORM-7) d ppm 8.03 (s, 1 H) 7.64 (dd, 7=7.89, 1.65 Hz, 1 H) 7.50 - 7.57 (m, 3 H) 7.31 - 7.39 (m, 1 H) 7.09 (t, 7=1.83 Hz, 1 H) 3.70 (t, 7=6.79 Hz, 2 H) 3.27 (t, 7=7.58 Hz, 2 H) 2.53 (quin, 7=7.18 Hz, 2 H)

LCMS (ESI+): m/z 418.9 (M+H). EXAMPLE 14. Synthesis of Compound 130

Scheme: Synthesis of 2-chloro-N-(3-chloro-5-cyanophenyl)-5-(l,l-dioxidoisothiazol idin-2- yl)benzamide 4. (Compound 130):

Experimental Procedures:

Preparation of compound 2:

To a solution of 5-amino-2-chloro-benzoic acid (15 g, 87.4 mmol, 1 eq) in 150 mL of methanol was added dropwise SOCh (15.6 g, 131.1 mmol, 9.5 mL, 1.5 eq) at 0°C. The mixture was stirred at 65°C for 12 hours and the mixture was concentrated in vacuum to give 19.4 g of crude compound 2 (HC1 salt) as a white solid, which was used into the next step directly.

Preparation of compound 3:

To a solution of methyl 5-amino-2-chloro-benzoate (5 g, 22.5 mmol, 1 eq, HC1 salt) in 50 mL of dichloromethane was added TEA (6.8 g, 67.6 mmol, 9.4 mL, 3 eq) and 3-chloropropane- 1-sulfonyl chloride (5.2 g, 29.3 mmol, 3.6 mL, 1.3 eq). The mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated under reduced pressure to give a residue, which was dissolved in 50 mL of methanol and then TEA (6.8 g, 67.6 mmol, 9.4 mL, 3 eq) was added. The mixture was stirred at 65 °C for 3 hours and then the mixture was concentrated under reduced pressure to remove methanol. The residue was diluted with 30 mL of ethyl acetate, washed three times with 30 mL of sat. NH4CI, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 6.5 g of crude compound 3 as a yellow solid, which was used into the next step directly.

Preparation of 2-chloro-N-(3-chloro-5-cyanophenyl)-5-(l,l-dioxidoisothiazol idin-2- yl)benzamide (Compound 130):

To a solution of methyl 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoate (800 mg, 2.8 mmol, 1 eq ) and 3-amino-5-chloro-benzonitrile (463.4 mg, 3.0 mmol, 1.1 eq ) in 5 mL of toluene was added AlMe 3 (2 M, 1.7 mL, 1.2 eq) dropwise at 0°C under N2 atmosphere. The solution was stirred for 0.5 hours at 0°C, and then heated to 100 °C and stirred for 2.5 hours at 100°C. The reaction mixture was quenched by addition of 5 mL of MeOH and concentrated under reduced pressure to give a residue, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether :ethyl acetate = 10: 1 to 1 : 1) to give 407.6 mg of Compound 130 (915.6 pmol, 33.2% yield, 92.2% purity) as a light yellow solid.

J H NMR (400 MHz, DMSO-de) <S ppm 11.09 (s, 1 H) 8.11 (t, 7 = 1.83 Hz, 1 H) 8.05 (t, 7 = 1.47 Hz, 1 H) 7.81 - 7.84 (m, 1 H) 7.61 (d, 7 = 8.68 Hz, 1 H) 7.37 - 7.41 (m, 2 H) 3.79 (t, 7 = 6.48 Hz, 2 H) 3.58 (t, 7 = 7.34 Hz, 2 H) 2.40 - 2.45 (m, 2 H)

LCMS (ESI+): m/z 410.0 (M+H).

The following compounds were prepared analogously to compound Compound 130 using different amines:

2-chloro-N-(3-chlorophenyl)-5-(l,l-dioxidoisothiazolidin-2-y l)benzamide (Compound 104):

J H NMR (400 MHz, METHANOL-c/4) d ppm 7.86 (s, 1 H) 7.49 - 7.59 (m, 2 H) 7.40 - 7.44 (m,

2 H) 7.36 (t, 7=8.13 Hz, 1 H) 7.18 (br d, 7=6.97 Hz, 1 H) 3.84 (t, 7=6.48 Hz, 2 H) 3.49 (t, 7=7.46 Hz, 2 H) 2.55 (quin, 7=6.94 Hz, 2 H)

LCMS (ESI+): m/z 384.9 (M+H). 2-chloro-N-(3-chloro-5-fluorophenyl)-5-(l,l-dioxidoisothiazo lidin-2-yl)benzamide (Compound 115):

JH NMR (400 MHz, CHLOROF ORM-7) d ppm 8.30 (br s, 1 H) 7.39 - 7.55 (m, 5 H) 6.93 (br d, 7=8.19 Hz, 1 H) 3.80 (t, 7=6.54 Hz, 2 H) 3.42 (t, 7=7.46 Hz, 2 H) 2.59 (quin, 7=7.00 Hz, 2 H)

LCMS (ESI+): m/z 403.0 (M+H).

2-chloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(3-isopropy lphenyl)benzamide (Compound 108):

J H NMR (400 MHz, CHLOROFORM-7) d ppm 7.98 (br s, 1 H) 7.31 - 7.44 (m, 5 H) 7.23 (t, 7=7.82 Hz, 1 H) 6.98 (d, 7=7.70 Hz, 1 H) 3.68 (t, 7=6.54 Hz, 2 H) 3.30 (t, 7=7.52 Hz, 2 H) 2.85 (dt, 7=13.82, 6.91 Hz, 1 H) 2.46 (quin, 7=7.00 Hz, 2 H) 1.20 (d, 7=6.85 Hz, 6 H)

LCMS (ESI+): m/z 393.1 (M+H).

N-(3,5-bis(trifluoromethyl)phenyl)-2-chloro-5-(l,l-dioxid oisothiazolidin-2-yl)benzamide (Compound 116):

JH NMR (400 MHz, CHLOROFORM-7) d ppm 8.60 - 8.86 (m, 1 H) 8.09 (s, 2 H) 7.58 (br s, 1 H) 7.36 (br d, 7=19.93 Hz, 1 H) 7.23 - 7.31 (m, 2 H) 3.59 - 3.74 (m, 2 H) 3.22 - 3.35 (m, 2 H) 2.38 - 2.52 (m, 2 H)

LCMS (ESI+): m/z 487.0 (M+H). 2-chloro-N-(3,5-dichlorophenyl)-5-(l,l-dioxidoisothiazolidin -2-yl)benzamide (Compound 111):

J H NMR (400 MHz, METHANOL-^) d ppm 7.74 (d, 7= 1 .83 Hz, 2 H) 7.49 - 7.57 (m, 1 H) 7.37 - 7.47 (m, 2 H) 7.24 (t, 7=1.90 Hz, 1 H) 3.84 (t, 7=6.54 Hz, 2 H) 3.49 (t, 7=7.46 Hz, 2 H) 2.55

(quin, 7=6.97 Hz, 2 H)

LCMS (ESI+): m/z 418.9 (M+H).

2-chloro-N-(3-chloro-5-methylphenyl)-5-(l,l-dioxidoisothi azolidin-2-yl)benzamide (Compound 120):

J H NMR (400 MHz, CHLOROFORM-7) d ppm 8.13 (br s, 1 H) 7.56 (s, 1 H) 7.37 - 7.49 (m, 3 H) 7.32 (s, 1 H) 6.99 (s, 1 H) 3.77 (t, 7=6.48 Hz, 2 H) 3.39 (t, 7=7.52 Hz, 2 H) 2.55 (quin, 7=7.00 Hz, 2 H) 2.35 (s, 3 H)

LCMS (ESI+): m/z 399.0 (M+H).

2-chloro-N-(3-chloro-5-(trifluoromethyl)phenyl)-5-(l,l-di oxidoisothiazolidin-2- yl)benzamide (Compound 113):

JH NMR (400 MHz, CHLOROFORM-7) d ppm 8.44 (br s, 1 H) 7.98 (s, 1 H) 7.81 (s, 1 H) 7.51 (s, 1 H) 7.40 - 7.46 (m, 3 H) 3.79 (t, 7=6.54 Hz, 2 H) 3.41 (t, 7=7.46 Hz, 2 H) 2.57 (quin, 7=6.97 Hz, 2 H).

LCMS (ESI+): m/z 453.0 (M+H). 2-chloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(3-ethylphenyl )benzamide (Compound 109):

J H NMR (400 MHz, CHLOROFORM-7) d ppm 7.93 (br s, 1 H) 7.31 - 7.51 (m, 5 H) 7.14 - 7.29 (m, 1 H) 6.96 (d, 7=7.58 Hz, 1 H) 3.70 (t, 7=6.54 Hz, 2 H) 3.32 (t, 7=7.52 Hz, 2 H) 2.60 (q, 7=7.58 Hz, 2 H) 2.48 (quin, 7=7.00 Hz, 2 H) 1.19 (t, 7=7.58 Hz, 3 H)

LCMS (ESI+): m/z 379.0 (M+H).

N-([l,l'-biphenyl]-3-yl)-2-chloro-5-(l,l-dioxidoisothiazo lidin-2-yl)benzamide (Compound 110):

J H NMR (400 MHz, CHLOROF ORM-7) d ppm 8.28 (s, 1 H) 7.89 (s, 1 H) 7.57 - 7.69 (m, 3 H) 7.32 - 7.49 (m, 8 H) 7.32 - 7.49 (m, 1 H) 3.72 (br t, 7=6.48 Hz, 2 H) 3.35 (t, 7=7.46 Hz, 2 H) 2.50 (quin, 7=6.94 Hz, 2 H)

LCMS (ESI+): m/z 427.0 (M+H).

2-chloro-N-(3,4-dichlorophenyl)-5-(l,l-dioxidoisothiazoli din-2-yl)benzamide (Compound 112):

J H NMR (400MHz, CHLOROFORM-d) d ppm 8.07 (br s, 1H) 7.85 (d, J=2.1 Hz, 1H) 7.45 (s, 1H) 7.41 - 7.34 (m, 4H) 3.73 (t, J=6.5 Hz, 2H) 3.35 (t, J=7.5 Hz, 2H) 2.51 (quin, J=7.0 Hz, 2H)

LCMS (ESI+): m/z 418.9 (M+H) . 2-chloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(3-(trifluorom ethyl)phenyl)benzamide (Compound 114):

J H NMR (400MHz, DMSO-d6) d ppm 10.91 (s, 1H), 8.20 (s, 1H) 7.91 (br d, J=8.4 Hz, 1H) 7.64 - 7.57 (m, 2H) 7.49 (br d, J=7.7 Hz, 1H) 7.39 - 7.33 (m, 2H) 3.79 (t, J=6.5 Hz, 2H) 3.57 (t, J=7.4 Hz, 2H) 2.42 (quin, J=6.9 Hz, 2H)

LCMS (ESI+): m/z 419.0 (M+H).

2-chloro-N-(3,5-difluorophenyl)-5-(l,l-dioxidoisothiazoli din-2-yl)benzamide (Compound 137):

1H NMR (400MHz, CHLOROFORM-d) d ppm 8.26 (br s, 1H), 7.50 (s, 1H), 7.44 (d, J=1.3 Hz, 2H), 7.29 (s, 1H), 6.64 (tt, J=2.2, 8.9 Hz, 1H), 3.80 (t, J=6.5 Hz, 2H), 3.42 (t, J=7.5 Hz, 2H), 2.58 (quin, J=7.1 Hz, 2H)

LCMS (ESI+): m/z 387.0 (M+H).

2-chloro-N-(3,5-dimethylphenyl)-5-(l,l-dioxidoisothiazoli din-2-yl)benzamide (Compound 119):

J H NMR (400MHz, CHLOROFORM-d) d ppm 7.87 (br s, 1H), 7.43 - 7.33 (m, 3H), 7.20 (s, 2H), 6.76 (s, 1H), 3.72 (t, J=6.5 Hz, 2H), 3.33 (t, J=7.5 Hz, 2H), 2.49 (quin, J=7.0 Hz, 2H), 2.26 (s, 6H)

LCMS (ESI+): m/z 379.1 (M+H). 2-chloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(3-fluoro-5-me thylphenyl)benzamide

(Compound 138):

1H NMR (400MHz, CHLOROFORM-d) d ppm 8.06 (br s, 1H), 7.54 - 7.39 (m, 4H), 7.12 (s, 1H), 6.72 (br d, J=9.3 Hz, 1H), 3.81 (t, J=6.5 Hz, 2H), 3.42 (t, J=7.5 Hz, 2H), 2.58 (quin, J=7.0

Hz, 2H), 2.37 (s, 3H)

LCMS (ESI+): m/z 383.0 (M+H).

EXAMPLE 15. Synthesis of Compound 117

The amines of following compounds were made through a Suzuki reaction and followed by a hydrogenation reaction.

Scheme:

Experimental Procedures:

Preparation of compound 2:

To a solution of 3-bromoaniline (0.1 g, 581.3 pmol, 1 eq), 4,4,5,5-tetramethyl-2-(2- methylprop-l-enyl)-l,3,2-dioxaborolane (127.0 mg, 697.6 pmol, 1.2 eq) and K2CO3 (160.7 mg, 1.2 mmol, 2 eq) in 1 mL of dioxane and 0.1 mL of water was added Pd(dppf)Cl2 (42.5 mg, 58.1 pmol, 0.1 eq) and purged with N2 for 3 times. The mixture was stirred at 100°C for 12 hours and the reaction mixture was filtered, the filtrate was concentrated under reduced pressure to give a residue which was purified by prep-TLC (S1O2, Petroleum ether : Ethyl acetate = 3: 1) to give 120 mg of crude compound 2 as a yellow oil. Preparation of compound 3:

2 3

To a solution of 3-(2-methylprop-l-enyl) aniline (120 mg, 815.1 mmol, 1 eq) in 5 mL of methanol was added Pd/C (10%, 0.1 g) under N2 atmosphere. The suspension was degassed and purged with ¾ for 3 times. The mixture was stirred under ¾ (50 Psi) at 25 °C for 1 hours and then filtered and the filtrate was concentrated under reduced pressure to give 130 mg of crude compound 3 as a yellow oil.

Compounds 2 and 3 were then used for the ester amide exchange reaction to make the following two compounds.

2-chloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(3-isobutylphe nyl)benzamide (Compound 117):

J H NMR (400 MHz, CHLOROF ORM-7) d ppm 8.08 (s, 1 H) 7.36 - 7.41 (m, 2 H) 7.33 - 7.34 (m, 1 H) 7.26 - 7.31 (m, 2 H) 7.18 - 7.21 (m, 1 H) 6.88 (d, 7=7.58 Hz, 1 H) 3.63 (t, 7=6.54 Hz, 2 H) 3.27 (t, 7=7.52 Hz, 2 H) 2.39 - 2.43 (m, 3 H) 1.81 (dquin, 7=13.46, 6.75, 6.75, 6.75, 6.75 Hz, 2 H) 0.84 (d, 7=6.60 Hz, 6 H)

LCMS (ESI+): m/z 407.1 (M+H).

2-chloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(3-(2-methy lprop-l-en-l-yl)phenyl)benzamide (Compound 128)

1H NMR (400 MHz, CHLOROFORM-7) d ppm 8.05 (s, 1 H) 7.47 - 7.51 (m, 3 H) 7.40 - 7.45 (m, 2 H) 7.29 - 7.35 (m, 1 H) 7.03 - 7.07 (m, 1 H) 6.25 - 6.29 (m, 1 H) 6.10 - 6.13 (m, 1 H) 3.75 - 3.81 (m, 2 H) 3.37 - 3.43 (m, 2 H) 2.51 - 2.59 (m, 2 H) 1.89 - 1.92 (m, 6 H)

LCMS (ESI+): m/z 405.0 (M+H). The following compounds were prepared in an analogous manner:

2-chloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(6-isopropylpy ridin-2-yl)benzamide

(Compound 179):

JH NMR (400MHz, CHLOROFORM-d) d ppm 8.66 (br d, J=8.4 Hz, 1H) 8.17 (br t, J=7.9 Hz, 1H) 7.55 - 7.49 (m, 1H) 7.47 - 7.40 (m, 2H) 7.27 - 7.24 (m, 1H) 3.88 (br t, J=6.4 Hz, 2H) 3.47 - 3.34 (m, 3H) 2.54 (quin, J=6.7 Hz, 2H) 1.40 (d, J=7.1 Hz, 6H)

LCMS (ESI+): m/z 394.1 (M+H).

(E)-2-chloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(3-(pro p-l-en-l-yl)phenyl)benzamide (Compound 125):

J H NMR (400MHz, CHLOROFORM-d) d ppm 7.97 (br s, 1H) 7.56 (s, 1H) 7.43 - 7.34 (m, 4H) 7.22 (t, J=7.8 Hz, 1H) 7.07 (d, J=7.7 Hz, 1H) 6.37 - 6.29 (m, 1H) 6.26 - 6.16 (m, 1H) 3.71 (t, J=6.5 Hz, 2H) 3.32 (t, J=7.5 Hz, 2H) 2.48 (quin, J=7.0 Hz, 2H), 1.82 (dd, J=1.3, 6.4 Hz, 3H)

LCMS (ESI+): m/z 391.0 (M+H).

Compound 126:

J H NMR (400MHz, CHLOROFORM-d) d ppm 8.03 (br s, 1H), 7.52 - 7.39 (m, 5H), 7.32 - 7.27 (m, 1H), 7.01 (d, J=7.6 Hz, 1H), 3.79 (t, J=6.5 Hz, 2H), 3.40 (t, J=7.5 Hz, 2H), 2.66 - 2.51 (m,

4H), 1.68 - 1.64 (m, 2H), 0.96 (t, J=7.3 Hz, 3H)

LCMS (ESI+): m/z 393.0 (M+H). 2-chloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(4-isopropylpy ridin-2-yl)benzamide

(Compound 121):

1H NMR (400MHz, CHLOROFORM-d) d ppm 8.81 (d, J=1.5 Hz, 1H) 8.15 (d, J=6.2 Hz, 1H), 7.58 - 7.54 (m, 1H) 7.50 - 7.43 (m, 2H) 7.31 (dd, J=1.6, 6.2 Hz, 1H) 3.91 (t, J=6.6 Hz, 2H) 3.40

(t, J=7.5 Hz, 2H) 3.15 (spt, J=6.9 Hz, 1H) 2.55 (quin, J=7.0 Hz, 2H) 1.39 (d, J=7.0 Hz, 6H)

LCMS (ESI+): m/z 394.1 (M+H).

2-chloro-5-(l,l-dioxidoisothiazolidin-2-yl)-N-(4-(prop-l- en-2-yl)pyridin-2-yl)benzamide (Compound 127):

J H NMR (400 MHz, METH AN OL-74) d ppm 8.34 (d, 7=5.99 Hz, 1 H) 7.96 (s, 1 H) 7.55 - 7.60 (m, 2 H) 7.53 (d, 7=2.81 Hz, 1 H) 7.43 - 7.47 (m, 1 H) 5.87 (s, 1 H) 5.55 (d, 7=0.73 Hz, 1 H) 3.86 (t, 7=6.54 Hz, 2 H) 3.51 (t, 7=7.46 Hz, 2 H) 2.56 (quin, 7=6.97 Hz, 2 H) 2.24 (s, 3 H) LCMS (ESI+): m/z 392.0 (M+H).

EXAMPLE 16. Synthesis of Compound 165

The following amine 4 was synthesized as shown and used to make the final product listed.

Experimental Procedures:

Preparation of compound B:

To a solution of tetrahydropyran-4-carboxylic acid (1 g, 7.7 mmol, 1 eq) and 2- hydroxyisoindoline-l,3-dione (1.3 g, 7.7 mmol, 1 eq) in 10 mL of dichloromethane was added EDCI (1.6 g, 8.5 mmol, 1.1 eq) and DMAP (187.8 mg, 1.5 mmol, 0.2 eq). The mixture was stirred at 30°C for 12 hours and the reaction mixture was partitioned between 10 mL of water and 20 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give a residue, which was purified by column chromatography (Si(¾, eluting with a gradient of Petroleum ether : Ethyl acetate = 100: 1 to 6: 1) to give 1 g of compound B (3.6 mmol, 47.3% yield) as a white solid.

Preparation of compound 2:

2 To a solution of 3-chloro-5-iodo-benzoic acid (3 g, 10.6 mmol, 1 eq) in 30 mL of toluene was added dropwise DPPA (2.9 g, 10.6 mmol, 2.3 mL, 1 eq) and TEA (1.1 g, 10.6 mmol, 1.5 mL, 1 eq) at 25°C. After addition, the mixture was stirred at this temperature for 1 hour and 80°C for 1 hour. And then 2-methylpropan-2-ol (944.7 mg, 12.8 mmol, 1.2 mL, 1.2 eq) was added dropwise. The resulting mixture was stirred at 110°C for 12 hours. Then it was partitioned between 10 mL of sat. NaHCCL and 10 mL of ethyl acetate. The organic phase was separated, washed with 6 mL of sat. NaHCCL, 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of Petroleum ether : Ethyl acetate = 100: 1 to 50: 1) to give 2.6 g of compound 2 (7.8 mmol, 70.3 % yield) as a yellow oil.

Preparation of compound 3:

Ni(dtbbpy)Br2 (82.6 mg, 169.7 pmol, 0.2 eq) was added to a mixture of tert-butyl N-(3- chloro-5-iodo-phenyl)carbamate (300 mg, 848.5 pmol, 1 eq), (l,3-dioxoisoindolin-2-yl) tetrahydropyran-4-carboxylate (350.3 mg, 1.3 mmol, 1.5 eq) and Zn (111.0 mg, 1.7 mmol, 2 eq) in 1.2 mL of DMA under N2. Then the mixture was heated to 40°C for 12 hours. The reaction mixture was partitioned between 3 mL of water and 5 mL of ethyl acetate. The organic phase was separated, washed three times with 6 mL of water and 2 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give a residue, which was purified by prep- TLC (S1O2, eluting with Petroleum ether : Ethyl acetate = 3: 1) to give 120 mg of compound 3 (384.9 pmol, 45.4% yield) as a yellow oil.

Preparation of compound 4:

3 4 A mixture of tert-butyl N-(3-chloro-5-tetrahydropyran-4-yl-phenyl)carbamate (120 mg, 384.9 pmol, 1 eq ) and TFA (770.0 mg, 6.8 mmol, 0.5 mL, 17.6 eq ) in 1 mL of dichloromethane was degassed and purged with N2 for 3 times, and then the mixture was stirred at 25°C for 0.5 hours under N2 atmosphere. The reaction mixture was partitioned between 2 mL of NaOH (1M) and 6 mL of ethyl acetate. The organic phase was separated, washed with 4 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 80 mg of crude compound 4 as a yellow solid.

Compound 4 was used for an ester amide exchange reaction following the normal procedure to give (Compound 165).

2-chloro-N-(3-chloro-5-(tetrahydro-2H-pyran-4-yl)phenyl)- 5-(l,l-dioxidoisothiazolidin-2- yl)benzamide (Compound 165):

J H NMR (400 MHz, CHLOROF ORM-7) d ppm 8.22 (s, 1 H) 7.61 (s, 1 H) 7.49 (s, 1 H) 7.43 (s, 2 H) 7.40 (s, 1 H) 7.03 (s, 1 H) 4.05 - 4.11 (m, 2 H) 3.79 (t, 7=6.48 Hz, 2 H) 3.47 - 3.55 (m, 2 H) 3.40 (t, 7=7.52 Hz, 2 H) 2.71 - 2.81 (m, 1 H) 2.57 (quin, 7=7.03 Hz, 2 H) 1.75 - 1.82 (m, 1 H)

1.75 - 1.82 (m, 3 H)

LCMS (ESI+): m/z 469.0 (M+H).

2-chloro-N-(3-chloro-5-(tetrahydrofuran-3-yl)phenyl)-5-(l ,l-dioxidoisothiazolidin-2- yl)benzamide Compound 166 was prepared analogously by coupling the appropriate acid with 2-hydroxyisoindoline-l, 3-dione:

2-chloro-N-(3-chloro-5-(tetrahydrofuran-3-yl)phenyl)-5-(l,l- dioxidoisothiazolidin-2- yl)benzamide (Compound 166):

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 8.14 (br s, 1 H) 7.64 (s, 1 H) 7.51 (s, 1 H) 7.44 (s, 2 H) 7.40 (s, 1 H) 7.07 (s, 1 H) 4.04 - 4.14 (m, 2 H) 3.91 (q, /= 7.66 Hz, 1 H) 3.80 (t, /=6.54 Hz, 2 H) 3.75 (dd, /=8.44, 6.97 Hz, 1 H) 3.35 - 3.44 (m, 3 H) 2.58 (quin, /=7.00 Hz, 2 H) 2.35 - 2.44 (m, 1 H) 2.01 (dq, /=12.46, 7.83 Hz, 1 H)

LCMS (ESI+): m/z 455.0 (M+H).

EXAMPLE 17. Synthesis of Compound 167

Preparation of amine 3:

Step 1 : Preparation of compound 2:

A mixture of tert-butyl N-(3-chloro-5-iodo-phenyl)carbamate (400 mg, 1.1 mmol, 1 eq), morpholine (118.3 mg, 1.4 mmol, 119.5 pL, 1.2 eq), Pd2(dba)3 (518.0 mg, 565.6 pmol, 0.5 eq), Xantphos (327.3 mg, 565.6 pmol, 0.5 eq) and tBuOK (190.4 mg, 1.7 mmol, 1.5 eq) in 3 mL of dioxane was degassed and purged with N2 3 times, and then the mixture was stirred at 110°C for 12 hours under N2 atmosphere. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified prep-TLC (SiC , eluting with Petroleum ether : Ethyl acetate = 3: 1) to give 0.2 g of compound 2 (639.4 pmol, 56.5% yield) as a yellow solid.

Step 2: Preparation of compound 3:

A mixture of tert-butyl N-(3-chloro-5-morpholino-phenyl)carbamate (0.2 g, 639.4 pmol, 1 eq) and TFA (616.0 mg, 5.4 mmol, 400.0 pL, 8.5 eq) in 1 mL of dichloromethane was degassed and purged with N2 3 times, and then the mixture was stirred at 25 °C for 0.5 hours under N2 atmosphere. The reaction mixture was concentrated in vacuum to give a residue. To the residue in 3 mL of methanol was added Na2CC>3 solid to make the pH ~10 at 25 °C. The mixture was filtered and the filtrate was concentrated under reduced pressure to give 640 mg of compound 3 (632.0 pmol, 98.8% yield, 21% purity) as a yellow solid.

Compound 3 was used for an ester amide exchange reaction following the normal procedure to give 2-chloro-N-(3-chloro-5-morpholinophenyl)-5-(l,l-dioxidoisoth iazolidin-2- yl)benzamide. (Compound 167):

J H NMR (400 MHz, CHLOROF ORM-7) d ppm 8.09 (br d, 7=7.95 Hz, 1 H) 7.47 (br s, 1 H) 7.42 (br s, 2 H) 7.29 (s, 1 H) 7.06 (s, 1 H) 6.68 (s, 1 H) 3.82 - 3.88 (m, 4 H) 3.77 (br t, 7=6.30 Hz, 2 H) 3.40 (br t, 7=7.46 Hz, 2 H) 3.19 (br d, 7=4.28 Hz, 4 H) 2.56 (quin, 7=6.97 Hz, 2 H)

LCMS (ESI+): m/z 470.0 (M+H).

EXAMPLE 18. Synthesis of Compound 170

Preparation of amine 3

Step 1: Preparation of compound 2:

To a solution of tert-butyl N-(3-chloro-5-iodo-phenyl)carbamate (300 mg, 848.5 pmol, 1 eq) in 4 mL of DME was added 3-iodooxetane (249.8 mg, 1.4 mmol, 1.6 eq) and 4-tert-butyl-2- (4-tert-butyl-2-pyridyl)pyridine (34.2 mg, 127.3 pmol, 0.15 eq), bis(trimethylsilyl)silyl- trime thy 1- silane (211.0 mg, 848.5 pmol, 261.8pL, 1 eq), dichloronickel;l,2-dimethoxyethane (18.6 mg, 84.9 pmol, 0.1 eq), bis[3,5-difluoro-2-[5-(trifluoromethyl)-2- pyridyl]phenyl]iridium(l+);4-tert-butyl-2-4-tert-butyl-2-pyr idyl)pyridine;hexafluorophosphate (9.5 mg, 8.5 pmol, 0.01 eq) and Na2C(¾ (179.9 mg, 1.7 mmol, 2 eq). The mixture was degassed and purged with N2 for 3 times, and then the mixture was irradiated with 34 W blue light at 30°C for 24 hours under N2 atmosphere. The residue was quenched by addition 5 mL of water and 9 mL of ethyl acetate, the organic phase was separated and washed with 6 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with Petroleum ether:Ethyl acetate = 5: 1) to give 50 mg of compound 2 (176.2 pmol, 20.8% yield) as a colorless solid.

Step 2: Preparation of compound 3:

TFA (616.0 mg, 5.4 mmol, 0.4 mL, 30.7 eq) was added to a solution of tert-butyl N-[3- chloro-5-(oxetan-3-yl)phenyl]carbamate (50 mg, 176.2 m ihoI, 1 eq) in 2 mL of dichloromethane at 30°C. The mixture was stirred at 30°C for 0.5 hours. The pH of the reaction mixture was adjusted by adding saturated NaOH solution to pH~10 at 0°C, and extracted with 15 mL of dichloromethane. The combined organic layers were washed with 10 mL of brine, dried with anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 30 mg of crude compound 3 as a yellow solid.

Compound 3 was used for an ester amide exchange reaction following the normal procedure to give 2-chloro-N-(3-chloro-5-(oxetan-3-yl)phenyl)-5-(l,l-dioxidois othiazolidin- 2-yl)benzamide. (Compound 170):

J H NMR (400 MHz, DMSO-d6) d ppm 10.79 (s, 1 H) 7.76 - 7.81 (m, 1 H) 7.69 (s, 1 H) 7.58 (d, 7=8.60 Hz, 1 H) 7.34 - 7.38 (m, 1 H) 7.31 - 7.34 (m, 1 H) 7.22 (s, 1 H) 4.94 (dd, 7=8.27, 6.06 Hz, 2 H) 4.57 (t, 7=6.28 Hz, 2 H) 4.24 (quin, 7=7.39 Hz, 1 H) 3.78 (t, 7=6.50 Hz, 2 H) 3.57 (t, 7=7.39 Hz, 2 H) 2.37 - 2.45 (m, 2 H)

LCMS (ESI+): m/z 441.0 (M+H).

EXAMPLE 19. Synthesis of Compound 131

Preparation of amine 2:

2

To a solution of l-chloro-3-methoxy-5-nitro-benzene (100 mg, 533.1 m ihoI, 1 eq) in 2 mL of ethanol was added Fe (148.9 mg, 2.7 mmol, 5 eq) and NH4CI (285.2 mg, 5.3 mmol, 10 eq) and water (0.5 mL). The mixture was stirred at 80°C for 1 hour. Then it was filtered and partitioned between 5 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 95 mg of crude compound 2 as a yellow oil, which was used into the next step directly.

Then compound 2 was used in an ester amide exchange reaction following the usual procedure to give 2-chloro-N-(3-chloro-5-methoxyphenyl)-5-(l,l-dioxidoisothiaz olidin-2- yl)benzamide(Compound 131).

J H NMR (400 MHz, DMSO-d6) d ppm 10.68 (s, 1 H) 7.57 (d, J=8.80 Hz, 1 H) 7.44 (t, J=1.71 Hz, 1 H) 7.33 - 7.38 (m, 1 H) 7.30 - 7.33 (m, 1 H) 7.26 - 7.29 (m, 1 H) 6.79 - 6.81 (m, 1 H) 3.77 - 3.81 (m, 2 H) 3.77 (s, 3 H) 3.56 (t, J=7.34 Hz, 2 H) 2.37 - 2.47 (m, 3 H)

LCMS (ESI+): m/z 415.0 (M+H)

2-chloro-N-(3-cyano-5-methylphenyl)-5-(l,l-dioxidoisothiazol idin-2-yl)benzamide (Compound 134) was prepared analogously reducing the appropriate nitro compound followed by an ester amide exchange reaction.

1H NMR (400 MHz, CHLOROFORM-d) d ppm 8.21 - 8.29 (m, 1 H) 7.82 - 7.89 (m, 1 H) 7.61 - 7.71 (m, 1 H) 7.53 (s, 1 H) 7.45 (d, J=1.54 Hz, 2 H) 7.28 (br s, 1 H) 3.81 (t, J=6.50 Hz, 2 H) 3.42 (t, J=7.50 Hz, 2 H) 2.54 - 2.63 (m, 2 H) 2.42 (s, 3 H)

LCMS (ESI+): m/z 390.0 (M+H)

EXAMPLE 20. Synthesis of Compound 139

Scheme: Synthesis of 2-chloro-N-(3-chloro-5-methylphenyl)-5-(l ,l-dioxidoisothiazolidin-2-yl)- 3-methoxybenzamide 3 (Compound 139):

Experimental Procedures:

Preparation of compound 2:

1 2

A mixture of methyl 5-bromo-2-chloro-3-methoxy-benzoate (200 mg, 715.5 pmol, 1 eq), 1 ,2-thiazolidine 1, 1-dioxide (104.0 mg, 858.6 pmol, 1.2 eq), K3PO4 (227.8 mg, 1.1 mmol, 1.5 eq), L-proline (41.2 mg, 357.8 pmol, 0.5 eq) and Cul (68.14 mg, 357.8 pmol, 0.5 eq) in 3 mL of DMSO was degassed and purged with N2 for three times, and then the mixture was stirred at 110°C for 12 hours under N2 atmosphere. It was then partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of water, 15mL of brine, dried over Na2S04, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- TLC(Si0 2 , eluting with a gradient of petroleum ether:ethyl acetate =5: 1) to give 30 mg of compound 2 (93.8 pmol, 13.1 % yield) as a yellow oil. Preparation of 2-chloro-N-(3-chloro-5-methylphenyl)-5-(l,l-dioxidoisothiazo lidin-2-yl)-3- methoxybenzamide 3 (Compound 139):

To a solution of compound 2 (28.7 mg, 89.9 mihoI, 1 eq) in 1 mL of toluene was added 3- chloro-5-methyl-aniline (15.3 mg, 107.9 mihoI, 1.2 eq) and AlMe3 (2 M, 67.41 pL, 1.5 eq) at 0°C. The mixture was stirred at 110°C for 4 hours and was quenched by addition of 1 mL of methanol, and concentrated under reduced pressure to give a residue, which was purified by prep- HPLC (basic condition) to give a crude product, which was then re -purified by prep- TLC (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =1 : 1) to give 6.5 mg of 2-chloro- N-(3-chloro-5-methylphenyl)-5-( 1 , 1 -dioxidoisothiazolidin-2-yl)-3-methoxybenzamide 3

(Compound 139) (14.9 mihoI, 16.5% yield, 98.1% purity) as a white solid.

J HNMR (400MHz, CHLOROFORM-d) d ppm 7.90 (s, 1H) 7.48 (s, 1H) 7.24 (s, 1H) 7.07 (d, J=2.6 Hz, 1H) 6.94 - 6.86 (m, 2H) 3.91 - 3.84 (m, 3H) 3.71 (t, J=6.5 Hz, 2H) 3.34 (t, J=7.5 Hz, 2H) 2.53 - 2.44 (m, 2H) 2.27 (s, 3H)

LCMS (ESI+): m/z 429.0 (M+H).

EXAMPLE 21. Synthesis of Compound 141

Scheme: Synthesis of 2-chloro-N-(3-chloro-5-methylphenyl)-5-(l,l-dioxidoisothiazo lidin-2-yl)- 3-fluorobenzamide 6 (Compound 141):

Experimental Procedures:

Preparation of compound 2:

1 2

To a solution of 2-chloro-3-fluoro-benzoic acid (1 g, 5.7 mmol, 1 eq) in H2SO4 (9.2 g,

91.9 mmol, 5 mL, 98% purity, 16.1 eq) was added HNO3 (7.0 g, 75.5 mmol, 5 mL, 68% purity, 13.2 eq) at 0°C. After addition, the mixture was stirred at this temperature for 0.5 hour. The resulting mixture was stirred at 25°C for 2.5 hours. Then the reaction as quenched by addition dropwise of 20 mL of ice-water, and extracted three times with 30 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 1.7 g of crude compound 2 as a yellow oil, which was used into the next step without further purification. Preparation of compound 3:

To a solution of 2-chloro-3-fluoro-5-nitro-benzoic acid (1.2 g, 5.5 mmol, 1 eq) in 6 mL of MeOH was added SOCh (975.4 mg, 8.2 mmol, 594.7 pL, 1.5 eq) at 0°C. The mixture was stirred at 65°C for 2 hours. Then it was partitioned between 20 mL of water and 50 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over Na 2 S0 4 , filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of Petroleum ether:Ethyl acetate = 1:0 to 50:1) to give 220 mg of compound 3 (941.9 pmol, 17.2% yield) as a white solid.

Preparation of compound 4:

To a solution of methyl 2-chloro-3-fluoro-5-nitro-benzoate (82 mg, 351.1 pmol, 1 eq) in 2 mL of toluene was added 3-chloro-5-methyl-aniline (49.7 mg, 351.1 pmol, 1 eq) and AlMe3 (2 M, 263.3 uL, 1.5 eq) at 0°C. The mixture was stirred at 100°C for 3 hours. Then it was added 2 ml of MeOH and concentrated under reduced pressure. Then it was partitioned between 5 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2S04, filtered and concentrated under reduced pressure to give 150 mg of crude compound 4 as a yellow solid, which was used into the next step without further purification.

Preparation of compound 5:

To a solution of 2-chloro-N-(3-chloro-5-methyl-phenyl)-3-fluoro-5-nitro-benza mide (150 mg, 437.1 pmol, 1 eq) in 6 mL of EtOH and 1.5 mL of ¾0 were added Fe (122.1 mg, 2.2 mmol, 5 eq) and NH4CI (233.8 mg, 4.4 mmol, 10 eq). The mixture was stirred at 80°C for 1 hour. Then it was filtered and partitioned between 5 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 120 mg of crude compound 5 as a yellow solid, which was used into the next step without further purification.

Preparation of 2-chloro-N-(3-chloro-5-methylphenyl)-5-(l,l-dioxidoisothiazo lidin-2-yl)-3- fluorohenz amide 6 (Compound 141):

To a solution of 5-amino-2-chloro-N-(3-chloro-5-methyl-phenyl)-3-fluoro-benza mide (120 mg, 383.2 pmol, 1 eq) in 2 mL of dichloromethane was added TEA (58.2 mg, 574.8 pmol, 1.5 eq) and 3-chloropropane-l-sulfonyl chloride (67.9 mg, 383.2 pmol, 1 eq) at 0°C. The mixture was stirred at 25°C for 3 hours and was concentrated under reduced pressure to give a residue. To the mixture was added 2.5 mL of MeOH and TEA (58.2 mg, 574.8 pmol, 1.5 eq) and the mixture was stirred at 65°C for 9 hours. It was then filtered, the filtrate was purified by prep-HPLC (TFA condition) to give 36.8 mg of compound 2-chloro-N-(3-chloro-5- methylphenyl)-5-(l,l-dioxidoisothiazolidin-2-yl)-3-fluoroben zamide 5 (Compound 141) (88.2 pumol, 23.0% yield, 100.0% purity) as a light yellow solid.

J H NMR (400 MHz, CHLOROFORM-d) d ppm 7.91 - 7.99 (m, 1 H) 7.53 - 7.58 (m, 1 H) 7.27 - 7.36 (m, 3 H) 6.98 - 7.03 (m, 1 H) 3.79 (t, J=6.54 Hz, 2 H) 3.40 - 3.46 (m, 2 H) 2.54 - 2.63 (m, 2 H) 2.36 (s, 3 H)

LCMS (ESI+): m/z 417.0 (M+H).

EXAMPLE 22. Synthesis of Compound 149

Preparation of 2,6-dichloro-N-(3,5-dichlorophenyl)-3-(l,l-dioxidoisothiazol idin-2-yl)benzamide

6 (Compound 149):

Experimental Procedures:

Preparation of compound 2: To a solution of compound 1 (1 g, 4.2 mmol, 1 eq) and DMF (15.5 mg, 211.9 pmol, 0.05 eq) in 10 mL of dichloromethane was added oxalyl chloride (1.1 g, 8.5 mmol, 741.8 pL, 2 eq) drop-wise at 0°C over 0.25 hour. The solution was stirred for 0.5 hour at 0°C and was warmed to 20°C and stirred for 1 hour. Anhydrous methanol (1.4 g, 42.4 mmol, 1.7 mL, 10 eq) was added drop-wise to the mixture at 20°C over 0.25 hour. The mixture was concentrated under reduced pressure at 40°C to give 1 g of crude compound 2 as a light yellow oil, which was used directly into the next step without purification.

Preparation of compound 4:

The solution of compound 2 (990 mg, 4.0 mmol, 1 eq) and compound 3 (769.8 mg, 4.8 mmol, 1.2 eq) in 10 mL of toluene was cooled to 0°C under N2. To the solution was added

AlMe3 (2 M, 3.0 mL, 1.5 eq) drop-wise at 0°C and the reaction mixture was stirred for 0.5 hour and then heated to 100°C. The reaction mixture was stirred for another 2.5 hours at 100°C, cooled to 20-25 °C and filtered, and the filtrate was partitioned between 20 mL of water and 10 mL of ethyl acetate. The organic phase was separated and the aqueous layer was extracted with 10 mL of ethyl acetate again. The organic layers were combined and concentrated under reduced pressure at 45 °C to give 2.2 g of crude compound 4 as red brown oil, which was used directly into the next step without purification.

Preparation of compound 5:

Compound 4 (2.2 g, 5.8 mmol, 1 eq) and NH4CI (3.1 g, 57.9 mmol, 10 eq) was dissolved in 20 mL of ethanol and 5 mL of water to give a red reaction mixture at 20°C. The mixture was heated to 70°C and Fe (3.2 g, 57.9 mmol, 10 eq) was added in 3 portions over 0.15 hour. The mixture was stirred for 12 hours at 70°C, the mixture was cooled to 20°C and filtered. The filtrate was concentrated under reduced pressure at 40°C to give a residue. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate = 10: 1 to 1 :1) to give 0.5 g of compound 5 (1.4 mmol, 24.7% yield) as a light yellow solid.

Preparation of compound 2,6-dichloro-N-(3,5-dichlorophenyl)-3-(l, l-dioxidoisothiazolidin-2- yl)benzamide 6 (Compound 149):

Compound 5 (100 mg, 285.7 pmol, 1 eq) and 3-chloropropane-l-sulfonyl chloride (101.2 mg, 571.4 pmol, 2 eq) was dissolved in 1 mL of dichloromethane to give a clear solution at 20°C. The reaction mixture was cooled to 0°C under N2, TEA (86.7 mg, 857.1 pmol, 119.3 pL,

3 eq) was added to the solution at 0°C and the mixture was warmed to 20°C and stirred for 12 hours. The mixture was concentrated under reduced pressure at 40°C to give a red residue. The residue and TEA (86.7 mg, 857.1 pmol, 119.3 pL, 3 eq) were dissolved in 2 mL of methanol and heated to 70°C. The mixture was stirred for another 2 hours at 70°C, cooled to 20°C and quenched with 1 mL of methanol. The quenched mixture was concentrated under reduced pressure at 40°C to give a residue. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate = 10: 1 to 1 : 1) to give 26 mg of compound 2,6-dichloro-N-(3,5-dichlorophenyl)-3-(l,l-dioxidoisothiazol idin-2-yl)benzamide 6 (Compound 149) (54.9 pmol, 19.2% yield, 95.8% purity) as a light yellow solid.

J H NMR (400MHz, DMSO-de) d ppm 11.25 (s, 1 H), 7.73 (dd, 7=1.4, 5.0 Hz, 4 H), 7.41 (t, 7=1.8 Hz, 1 H), 3.71 (br d, 7=18.7 Hz, 2 H), 3.46 (t, 7=7.4 Hz, 2 H), 2.47 - 2.43 (m, 2 H).

LCMS (ESI+): mJz 452.9 (M+H).

EXAMPLE 23: Synthesis of compound 214:

Experimental Procedures:

Preparation of compound 2:

To a solution of l-chloro-3-methoxy-5-nitro-benzene (3 g, 15.9 mmol, 1 eq ) in 30 mL of dichloromethane was added BBb (8.0 g, 31.9 mmol, 3.1 mL, 2 eq) at 0°C. The mixture was stirred at 25°C for 12 hours. Then it was partitioned between 30 mL of water and 50 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =100:1 to 30: 1) to give 1.9g of compound 2 (10.9 mmol, 68.5% yield) as a pink solid. Preparation of compound 3:

To a solution of 3-chloro-5-nitro-phenol (1.9 g, 10.9 mmol, 1 eq) in 20 mL of ethyl alcohol and 5 mL of water was added Fe (3.1 g, 54.7 mmol, 5 eq) and NH4CI (5.9 g, 109.5 mmol, 10 eq). The mixture was stirred at 85°C for 1 hour. Then it was partitioned between 15 mL of water and 40 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the 1.2 g of crude compound 3 as a brown solid.

Preparation of compound 5:

To a solution of 3-amino-5-chloro-phenol (50 mg, 348.3 mihoI, 1 eq) in 3 mL of toluene was added methyl 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoate (100.9 mg, 348.3 mpioΐ, 1 eq) and AlMe3 (2 M, 261.2 pL, 1.5 eq) at 0°C. The mixture was stirred at 100°C for 12 hours. Then it was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =0:1) to give 120 mg of compound 5 ( 299.1 pmol, 85.9% yield) as a white solid.

Preparation of compound 214:

5 214 To a solution of 2-chloro-N-(3-chloro-5-hydroxy-phenyl)-5-(l,l-dioxo-l,2-thia zolidin - 2-yl)benzamide (50 mg, 124.6 pmol, 1 eq ) in 2 mL of dimethyl formamide was added K2CO3 (34.4 mg, 249.2 pmol, 2 eq) and bromocyclobutane (50.5 mg, 373.8 pmol, 3 eq). The mixture was stirred at 80°C for 5 hours. Then it was filtered and concentrated under reduced pressure to give a residue, which was purified by prep- HPLC (TFA condition) to give 11.4 mg of 214 (24.8 pmol, 19.9% yield, 99.0% purity) as a white solid.

J H NMR (400MHz, CHLOROFORM-d) d ppm 8.04 (br s, 1H), 7.48 - 7.38 (m, 3H), 7.18 (br d, J = 17.0 Hz, 2H), 6.63 (br s, 1H), 4.70 - 4.59 (m, 1H), 3.77 (br t, / = 5.8 Hz, 2H), 3.39 (br t, / =

7.2 Hz, 2H), 2.62 - 2.41 (m, 4H), 2.23 - 2.09 (m, 2H), 1.93 - 1.80 (m, 1H), 1.76 - 1.61 (m, 1H). LCMS (ESI+): m/z 455.0 (M+H)

Compound 210 was prepared analogously using 3-bromooxetane to do alkylation at the last step:

compound 210

JH NMR (400MHz, CHLOROFORM-d) d ppm 8.30 (br s, 1H), 7.41 (br s, 3H), 7.23 (br s, 1H), 7.08 (br s, 1H), 6.53 (br s, 1H), 5.18 (br s, 1H), 4.95 (br s, 2H), 4.74 (br s, 2H), 3.76 (br s, 2H), 3.36 (br t, / = 6.7 Hz, 2H), 2.61 - 2.49 (m, 2H)

LCMS (ESI+): m/z 457.0 (M+H)

EXAMPLE 24: Preparation of Compound 231

Compound 231 was prepared through three methods:

Method A: (batch 1)

Experimental Procedures:

Preparation of compound 3:

To a solution of 3-chloro-5-nitro-phenol (50 mg, 288.1 pmol, 1 eq) in 2 mL of acetonitrile was added K2CO3 (79.6 mg, 576.2 pmol, 2 eq) and 4-bromotetrahydropyran (142.6 mg, 864.3 pmol, 3 eq). The mixture was stirred at 80°C for 36 hrs. Then KI (47.8 mg, 288.1 pmol, 1 eq) was added and stirred at 80°C for another 12 hrs. After reaction, it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =5: 1) to give 30 mg of compound 3 (116.4 pmol, 40.4% yield) as a yellow oil.

Preparation of compound 4:

To a solution of 4-(3-chloro-5-nitro-phenoxy)tetrahydropyran (30 mg, 116.4 pmol, 1 eq) in 2 mL of ethyl alcohol and 1 mL of water was added Fe (32.5 mg, 582.1 pmol, 5 eq) and NH4CI (31.1 mg, 582.14 mihoI, 5 eq). The mixture was stirred at 85°C for 12 hours. After reaction, it was filtered, the filtrate was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 30 mg of crude compound 4 as a yellow oil, which was used into the next step directly.

Preparation of compound 231 (batch 1):

4 231

To a solution of 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoic acid (43.6 mg, 158.1 m mol, 1.2 eq) in 2 mL of dimethyl form a ide was added 3-chloro-5-tetrahydropyran-4-yloxy- aniline (30 mg, 131.8 m mol, 1 eq), TEA (14.7 mg, 144.9 mpioΐ, 1.1 eq). The mixture was cooled to 0°C, HATU (55.1 mg, 144.9 mpioΐ, 1.1 eq) was added and stirred at 15°C for 12 hrs. The reaction mixture was filtered and concentrated under reduced pressure to give a residue, which was purified by prep- HPLC (TFA condition) to give 7.6 mg of 231 (15.6 mihoI, 11.8% yield, 99.4% purity) as a white solid.

J H NMR (400MHz, CHLOROFORM-d) d ppm 8.09 (s, 1H), 7.48 (s, 1H), 7.43 (s, 2H), 7.32 (s, 1H), 7.19 (s, 1H), 6.74 (s, 1H), 4.51 (td, / = 3.8, 7.7 Hz, 1H), 4.02 - 3.94 (m, 2H), 3.78 (t, / = 6.5 Hz, 2H), 3.60 (ddd, / = 3.1, 8.3, 11.6 Hz, 2H), 3.40 (t, / = 7.5 Hz, 2H), 2.57 (quin, / = 7.0 Hz, 2H), 2.10 - 2.01 (m, 2H), 1.86 - 1.73 (m, 2H)

FCMS (ESI+): m/z 485.1 (M+H)

Compounds 219 and 228 were prepared analogously using appropriate halide to do alkylation at the first step:

compound 219 J H NMR (400MHz, CHLOROFORM-d) d ppm 8.04 (br s, 1H), 7.51 - 7.39 (m, 3H), 7.22 (br d, 7 = 8.9 Hz, 2H), 6.72 (s, 1H), 4.05 (q, 7 = 7.0 Hz, 2H), 3.78 (t, 7 = 6.5 Hz, 2H), 3.40 (t, 7 = 7.5 Hz, 2H), 2.57 (quin, 7 = 7.0 Hz, 2H), 1.42 (t, 7 = 7.0 Hz, 3H)

LCMS (ESI+): m/z 429.0 (M+H)

c

compound 228: TV Boc

J H NMR (400MHz, CHLOROFORM-7) d ppm 8.00 (s, 1H), 7.44 - 7.36 (m, 3H), 7.15 - 7.10 (m, 2H), 6.50 (t, 7 = 2.0 Hz, 1H), 4.86 - 4.79 (m, 1H), 4.86 - 4.79 (m, 1H), 4.25 (dd, 7 = 6.5, 9.6 Hz, 2H), 3.93 (dd, 7 = 3.9, 9.8 Hz, 2H), 3.73 (t, 7 = 6.5 Hz, 2H), 3.34 (t, 7 = 7.5 Hz, 2H), 2.50 (quin, 7 = 6.9 Hz, 2H), 1.38 (s, 9H)

LCMS (ESI+): m/z 578.1 (M+Na)

Method B: (batch 2)

Experimental Procedures:

Preparation of compound 3:

To a solution of l-chloro-3-fluoro-5-nitro-benzene (100 mg, 569.6 mihoI, 1 eq) in 2 mL of dimethyl form amide was added NaH (34.2 mg, 854.5 mihoI, 60% purity, 1.5 eq) at 0°C. The mixture was stirred at 0°C for 0.5 hour. Then tetrahydropyran-4-ol (69.8 mg, 683.6 mihoI, 68.5 pL, 1.2 eq) was added and stirred at 20°C for 11.5 hours. After reaction, it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 150 mg of crude compound 3 as a yellow oil, which was used directly into the next step.

Preparation of compound 4:

To a solution of 4-(3-chloro-5-nitro-phenoxy)tetrahydropyran (150 mg, 582.1 mihoI, 1 eq) in 5 mL of ethyl alcohol and 1 mL of water was added Fe (162.5 mg, 2.9 mmol, 5 eq) and NH4CI (155.7 mg, 2.9 mmol, 5 eq). The mixture was stirred at 80°C for 1 hour. Then it was filtered, the filtrate was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 90 mg of crude compound 4 as a yellow oil, which was used into the next step directly.

Preparation of compound 231

To a solution of 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoic acid (100 mg, 362.7 mitioI, 1 eq) in 2 mL of dimethyl form amide was added compound 5 (82.6 mg, 362.7 mitioI, 1 eq) and TEA (44.0 mg, 435.2 miti I, 1.2 eq). The mixture was cooled to 0°C, HATU (165.5 mg, 435.2 mitioI, 1.2 eq) was added and then stirred at 25 °C for 12 hours. After reaction, the reaction mixture was filtered, the filtrate was purified twice by prep- HPLC (TFA condition) to give 25.4 mg of 231 (51.6 pmol, 14.2% yield, 98.6% purity) as a white solid.

J H NMR (400MHz, DMSO-r/6) d ppm 10.69 (br s, 1H), 7.58 (br d, 7=8.0 Hz, 1H), 7.43 (br s, 1H), 7.38 - 7.23 (m, 3H), 6.87 (br s, 1H), 4.58 (br s, 1H), 3.80 (br d, 7=19.8 Hz, 4H), 3.60 - 3.47 (m, 4H), 2.43 - 2.37 (m, 2H), 1.95 (br s, 2H), 1.59 (br s, 2H)

LCMS (ESI+): m/z 485.0 (M+H)

Method C: (batch 3, scale up reaction)

Experimental Procedures:

Preparation of compound 3:

To a solution of 3-chloro-5-nitro-phenol (12 g, 69.1 mmol, 1 eq ) in 120 mL of dichloromethane was added PPI13 (21.7 g, 82.9 mmol, 1.2 eq) and tetrahydropyran-4-ol (7.8 g, 76.1 mmol, 7.6 mL, 1.1 eq). Then DIAD (16.8 g, 82.9 mmol, 16.1 mL, 1.2 eq) was added at

0°C. The mixture was stirred at 25°C for 12 hours. Then it was partitioned between 500 mL of 1M NaOH and 1000 mL of ethyl acetate. The organic phase was separated, washed with 500 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =10:1 to 5:1) to give 12 g of compound 3 (46.6 mmol, 67.4% yield) as a yellow oil.

Preparation of compound 4:

To a solution of 4-(3-chloro-5-nitro-phenoxy)tetrahydropyran (14 g, 54.3 mmol, 1 eq) in 80 mL of ethyl alcohol and 10 mL of water was added Le (15.2 g, 271.7 mmol, 5 eq) and NH4CI (14.5 g, 271.7 mmol, 5 eq). The mixture was stirred at 80 °C for 12 hours. After reaction, it was filtered, the filtrate was concentrated under reduced pressure to give a residue, which was partitioned between 500 mL of water and 1L of ethyl acetate. The organic phase was separated, washed with 500 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =10:1 to 5:1) to give 8.8 g of compound 4 (38.6 mmol, 71.1% yield) as a yellow oil.

Preparation of compound 231 (batch 3):

To a solution of 3-chloro-5-tetrahydropyran-4-yloxy-aniline (20 g, 87.8 mmol, 1 eq) in 500 mL of dimethyl form amide was added compound 5 (24.2 g, 87.8 mmol, 1 eq) and TEA (10.7 g, 105.4 mmol, 14.7 mL, 1.2 eq). The mixture was cooled to 0°C, HATU (40.1 g, 105.4 mmol, 1.2 eq) was added and stirred at 25°C for 5 hours. After reaction, the mixture was added dropwise into 400 mL of water with stirring. The resulting mixture was filtered and the filter cake was washed with 50 mL of methanol, and dried in vacuum to give a crude product. The crude product was dissolved in 500 mL of dichloromethane at 50°C and stirred at 80°C for 12 hours. Then the mixture was cooled to 25 °C with stirring for 0.5 hour. The mixture was filtered and the filter cake was dried in vacuum to get a off-white solid, which was added 500 mL of water and stirred at 100°C for another 12 hours. Then the mixture was filtered and the filter cake was washed for three times with 300 mL of water, dried in the vacuum oven at 80°C over 6 hours to give 22 g of crude product, which was then dissolved in 400 mL of dichloromethane and 400 mL of methanol at 50°C. The solution was stirred at 70°C for 0.5 hour and then concentrated under reduced pressure to give a residue, which was added 2 L of methanol and stirred at 80°C for 6 hours. The resulting mixture was concentrated under reduced pressure to give a residue, which was added into 2 L of water and stirred at 110°C for 10 hours. Then the mixture was filtered and dried in the vacuum oven to give 20.3 g of 231 (41.7 mmol, 91.9% yield, 99.5% purity) as a off-white solid.

J H NMR (400MHz, DMSO-de) d ppm 10.66 (s, 1H), 7.57 (d, / = 8.8 Hz, 1H), 7.43 (t, / = 1.6 Hz, 1H), 7.38 - 7.25 (m, 3H), 6.86 (t, / = 1.9 Hz, 1H), 4.57 (tt, / = 4.1, 8.6 Hz, 1H), 3.89 - 3.74 (m, 4H), 3.56 (t, / = 7.4 Hz, 2H), 3.51 - 3.42 (m, 2H), 2.42 (quin, / = 6.9 Hz, 2H), 2.01 - 1.92 (m, 2H), 1.64 - 1.52 (m, 2H)

LCMS (ESI+): m/z 485.0 (M+H)

The following compounds were prepared analogously using the appropriate alcohol to do Mitsunobu Reaction, followed by reduction of nitro group and amide coupling reaction:

J H NMR (400MHz, CHLOROFORM-d) d ppm 8.06 (br s, 1H), 7.49 - 7.41 (m, 3H), 7.30 (s,

1H), 7.18 (s, 1H), 6.72 (s, 1H), 4.53 - 4.45 (m, 1H), 3.78 (t, / = 6.6 Hz, 2H), 3.72 - 3.62 (m, 2H), 3.44 - 3.30 (m, 4H), 2.56 (quin, 7 = 7.0 Hz, 2H), 1.96 - 1.88 (m, 2H), 1.81 - 1.69 (m, 2H), 1.47 (s, 9H)

LCMS (ESI+): m/z 429.0 (M+H)

compound 237

JH NMR (400MHz, CHLOROFORM-7) d ppm 8.02 (br s, 1H), 7.44 - 7.32 (m, 3H), 7.22 - 7.16 (m, 2H), 6.61 (t, 7=1.9 Hz, 1H), 4.86 (br d, 7 = 1.2 Hz, 1H), 3.95 - 3.80 (m, 4H), 3.71 (t, 7 = 6.5 Hz, 2H), 3.32 (t, 7 = 7.5 Hz, 2H), 2.49 (quin, 7 = 7.0 Hz, 2H), 2.21 - 2.04 (m, 2H)

LCMS (ESI+): m/z 471.0 (M+H)

compound 274:

JH NMR (400MHz, CHLOROFORM-7) d ppm 8.16 (br s, 1H), 7.37 - 7.30 (m, 3H), 7.15 (br s, 2H), 6.60 (s, 1H), 4.85 (br s, 1H), 3.95 - 3.81 (m, 4H), 3.68 (t, 7 = 6.4 Hz, 2H), 3.34 - 3.25 (m, 2H), 2.47 (quin, 7 = 6.9 Hz, 2H), 2.20 - 2.04 (m, 2H)

LCMS (ESI+): m/z 471.0 (M+H)

compound 279

1H NMR (400 MHz, CHLOROFORM-7) d ppm 8.09 (br s, 1 H) 7.57 (br s, 1 H) 7.52 (br s, 2 H) 7.33 (br d, 7 = 4.65 Hz, 2 H) 6.76 (br s, 1 H) 5.02 (br s, 1 H) 3.95 - 4.13 (m, 4 H) 3.83 - 3.90 (m, 2 H) 3.45 - 3.51 (m, 2 H) 2.60 - 2.69 (m, 2 H) 2.19 - 2.36 (m, 2 H)

LCMS (ESI+): m/z 471.0 (M+H)

Experimental Procedures:

The synthesis of compound 1 can be found in the report of 224, 225.

Preparation of compound 2:

1 3

To a solution of compound 1 (400 mg, 861.8 pmol, 1 eq ) compound 2 (346.4 mg, 1.1 mmol, 1.3 eq) in 4 mL of dioxane and 1 mL of water was added K2CO3 (238.2 mg, 1.7 mmol, 2 eq) and Pd(dppf)Cl2 (63.1 mg, 86.2 pmol, 0.1 eq) under N2, then the mixture was stirred at 100°C for 10 hours. After reaction, it was added 10 mL of water, extracted three times with 9 mL of ethyl acetate. The combined organic layers was washed with 12 mL of brine, dried with anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate=l:0 to 1:1) to give 320 mg of compound 3 ( 564.9 pmol, 65.6% yield) as a yellow solid. Preparation of compound 197:

To a solution of compound 3 (320 mg, 564.9 pmol, 1 eq ) in 2 mL of ethyl acetate was added Pd/C (50 mg, 10% purity) under N2, the suspension was degassed under vacuum and purged with ¾ several times. The mixture was stirred at 20°C for 2 hours under ¾ (15psi). After reaction, it was filtered and the filtrate was concentrated under reduced pressure to give 250 mg of crude 197 as a yellow solid, which was used directly into the next step.

Preparation of compound 223:

A solution of 197 (230 mg, 404.6 pmol, 1 eq) in 2 mL of 4M hydrochloric acid/ethyl acetate (19.8 eq) was stirred at 20°C for 20 min. After reaction, the suspension was filtered and the filtrate was concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (TFA condition) to give 5.6 mg of 223 (8.4 pmol, 2.1% yield, 87.8 % purity, TFA salt) as a white solid.

J H NMR (400 MHz, METH AN OL-dA) d ppm 7.68 (s, 1 H) 7.56 (t, / =1.83 Hz, 1 H) 7.51 (d, J =8.80 Hz, 1 H) 7.42 (d, / =2.69 Hz, 1 H) 7.35 - 7.39 (m, 1 H) 7.11 (t, / =1.53 Hz, 1 H) 3.82 (t, / =6.48 Hz, 2 H) 3.46 - 3.53 (m, 4 H) 3.09 - 3.21 (m, 2 H) 2.94 (tt, / =12.03, 3.32 Hz, 1 H) 2.53 (quin, / =6.97 Hz, 2 H) 2.12 (br d, J =14.06 Hz, 2 H) 1.82 - 1.98 (m, 2 H)

LCMS (ESI+): m/z 468.0 (M+H) EXAMPLE 26: Synthesis of compounds 224, 225:

To a solution of 3-bromo-5-chloro-aniline (3.6 g, 17.3 mmol, 1 eq) and compound 1 (5 g, 17.3 mmol, 1 eq) in 50 mL of toluene was added AlMe3 (2M, 12.9 mL, 1.5 eq) at 0°C. The mixture was stirred at 110°C for 3 hours. Then it was partitioned between 30 mL of water and 100 mL of ethyl acetate. The organic phase was separated, washed with 20 mL of brine, dried with anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate

=10: ltol :5) to give 4.2 g of compound 2 ( 9.1 mmol, 52.4% yield) as a yellow solid.

Preparation of compound 4:

To the solution of compound 2 (1 g, 2.2 mmol, 1 eq ), compound 3 (656.5 mg, 2.6 mmol, 1.2 eq) and AcONa (530.2 mg, 6.5 mmol, 3 eq) in 10 mL of dioxane was added Pd(dppf)Ch (157.6 mg, 215.4 pmol, 0.1 eq) under N2 at 20°C. Then the mixture was heated to 70°C and stirred for 12 hours. After reaction, it was cooled to 20°C and poured into 20 mL of water. The aqueous phase was extracted with 10 mL of ethyl acetate, separated and the organic layer, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate=10: l to 1 : 1) to give 400 mg of compound 4 ( 932.2 pmol, 43.3% yield) as a light yellow solid.

Preparation of compound 5:

5A 5

To a solution of tetrahydrofuran-3-one (1 g, 11.6 mmol, 1 eq) in 10 mL of THF was added LiHMDS (1 M, 17.4 mL, 1.5 eq) at -78°C under N2. The mixture was stirred at 0°C for 30 min, then l,l, l-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfo namide (6.2 g, 17.4 mmol, 1.5 eq) in 5 mL of THF was added dropwise. The mixture was stirred at 0°C for another 1.5 hrs. After reaction, it was quenched by addition of 20 mL of water at 0°C, then the aqueous phase was extracted three times with 9 mL of ethyl acetate. The combined organic layers was washed twice with 12 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, Petroleum ether/Ethyl acetate=l/0 to 20:1) to give 1.3 g of crude compound 5 as a white solid, which was used directly into the next step.

Preparation of compound 6:

To a solution of compound 4 (100 mg, 233.1 pmol, 1 eq), compound 5 (50.8 mg, 233.1 pmol, 1 eq) in 3 mL of dioxane and 0.5 mL of water was added K2CO3 (64.4 mg, 466.1 pmol, 2 eq) and Pd(dppf)Cl2 (17.1 mg, 23.3 pmol, 0.1 eq) under N2, then the mixture was stirred at 110°C for 10 hours. After reaction, it was added 10 mL of water, the aqueous phase was extracted three times with 9 mL of ethyl acetate. The combined organic phase was washed twice with 12 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by prep-TLC (S1O2, eluting with petroleum ether:ethyl acetate=0:l) to give 60 mg of compound 6 (132.4 pmol, 56.8% yield) as a yellow solid.

Preparation of compound 224 and 225:

To a solution of compound 6 (60 mg, 132.6 pmol, 1 eq) in 2 mL of ethyl acetate was added Pd/C (20 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with ¾ several times. Then mixture was stirredat 20°C for 10 min under H2 (15 psi) atmosphere. After reaction, the suspension was filtered and the combined filtrates were concentrated under reduced pressure to give a residue, which was purified by prep- HPLC (TFA condition) to give 9 mg of racemic product as a white solid. Then it was further separated by SFC (Method: 5-40_4_4min,5 pL,IPA(0.05%IPAm), column: DAICEL CHIRALPAK

IC(250mm*30mm,10pm);mobile phase: [0.1%NH3WATER IPA];B%: 42%-42%,25min) to give 2 mg of 224 ( 4.4 m ihoI, 3.3% yield, 100% purity) ) and 2.1 mg of 225(4.6 mihoI, 3.5% yield, 100% purity) both as white solid.

The structures of the two isomers were random assignment.

224: J H NMR (400 MHz, DMSO-de) d ppm 10.72 (s, 1 H) 7.76 (s, 1 H) 7.57 (d, 7 =8.82 Hz, 1 H)

7.52 (s, 1 H) 7.33 - 7.39 (m, 1 H) 7.31 (d, 7 =2.87 Hz, 1 H) 7.11 (s, 1 H) 3.90 - 4.05 (m, 2 H) 3.73 - 3.84 (m, 3 H) 3.56 (t, 7 =7.28 Hz, 3 H) 3.35 - 3.44 (m, 1 H) 2.38 - 2.45 (m, 2 H) 2.33 (dq, 7 =8.02, 4.01 Hz, 1 H) 1.88 (dq, 7 =12.29, 7.81 Hz, 1 H)

LCMS (ESI+): m/z 455.0 (M+H)

225:

J H NMR (400 MHz, DMSO-de) d ppm 10.72 (s, 1 H) 7.76 (s, 1 H) 7.58 (d, 7 =8.82 Hz, 1 H) 7.52 (s, 1 H) 7.34 - 7.38 (m, 1 H) 7.32 (d, 7 =2.65 Hz, 1 H) 7.11 (s, 1 H) 3.90 - 4.05 (m, 2 H) 3.73 - 3.83 (m, 3 H) 3.57 (t, 7 =7.28 Hz, 3 H) 3.36 - 3.44 (m, 1 H) 2.39 - 2.45 (m, 2 H) 2.29 - 2.36 (m, 1 H) 1.89 (dq, 7 =12.27, 7.82 Hz, 1 H)

LCMS (ESI+): m/z 455.0 (M+H)

EXAMPLE 27: Synthesis of compound 227:

Experimental Procedures:

The synthesis of compound 6 can be found in report of 224, 225.

Preparation of compound 3:

To a solution of 3-bromo-5-methyl-aniline (300 mg, 1.6 mmol, 1 eq) and compound 2 (513.9 mg, 1.8 mmol, 1.1 eq) in 5 mL of toluene was added AlMe3 (2 M, 1.2 mL, 1.5 eq) at 0°C The mixture was stirred at 110°C for 12 hours. Then it was partitioned between 5 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate=10: l to 2: 1) to give 458 mg of compound 2 (1.0 mmol, 64.0% yield) as a yellow oil.

Preparation of compound 5:

To a solution of compound 3 (450 mg, l. lmmol, 1 eq), compound 4 (309.0 mg, 1.2 mmol, 1.2 eq) and NaOAc (249.6 mg, 3.0 mmol, 3 eq) in 5 mL of dioxane was added

Pd(dppf)Cl2 (74.2 mg, 101.4 pmol, 0.1 eq) under N2 at 20°C. Then the mixture was heated to 100°C and stirred for 12 hours. After reaction, it was cooled to 20°C and poured into 20 mL of water. The aqueous phase was extracted with 10 mL of ethyl acetate, separated the organic layer, dried over Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate=10: l to 0: 1) to give 300 mg of compound 3 (611.2 pmol, 60.3% yield) as a yellow oil.

Preparation of compound 7:

To a solution of compound 5(150 mg, 305.6 pmol, 1 eq), compound 6 (266.7 mg, 1.2 mmol, 4 eq) and CS2CO3 (298.7 mg, 916.8 pmol, 3 eq) in 4 mL of dioxane and 1 mL of water was added Pd(dppf)Cl2 (22.4 mg, 30.6 pmol, 0.1 eq). The mixture was stirred at 110°C for 12 hours. Then it was partitioned between 10 mL of water and 10 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate=10: l to 1: 1) to give 20 mg of compound 7 (46.2 pmol, 15.1% yield) as a yellow solid.

Preparation of compound 227:

To a solution of compound 7 (20 mg, 46.2 pmol, 1 eq) in 2 mL of ethyl acetate was added Pd/C (20 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with ¾ several times. The mixture was stirred at 20°C for 10 min under ¾ (15 psi). The mixture was filtered and concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (TFA condition) to give 1.1 mg of 227 (2.5 pmol, 5.5% yield, 100% purity) as a white solid. J H NMR (400 MHz, DMSO-de) d ppm 10.46 - 10.40 (m, 1H), 7.57 - 7.53 (m, 1H), 7.44 - 7.41 (m, 2H), 7.36 - 7.31 (m, 1H), 7.30 - 7.27 (m, 1H), 6.87 - 6.84 (m, 1H), 4.05 - 3.98 (m, 1H), 3.97 - 3.90 (m, 1H), 3.82 - 3.75 (m, 3H), 3.58 - 3.51 (m, 3H), 3.30 - 3.27 (m, 1H), 2.45 - 2.39 (m, 2H), 2.34 - 2.29 (m, 1H), 2.29 - 2.27 (m, 3H), 1.95 - 1.82 (m, 1H)

LCMS (ESI+): m/z 435.1 (M+H)

The following compounds were prepared analogously coupling the appropriate trifluoromethanesulfonate with corresponding boronic acid firstly, followed by reduction of the double bond:

compound 236:

J H NMR (400 MHz, DMSO-de) d ppm 10.74 - 10.78 (m, 1 H) 7.79 (br s, 1 H) 7.59 - 7.64 (m, 1 H) 7.54 - 7.57 (m, 1 H) 7.38 - 7.42 (m, 1 H) 7.34 - 7.37 (m, 1 H) 7.15 - 7.18 (m, 1 H) 3.79 - 3.93 (m, 4 H) 3.61 (br t, 7 =7.24 Hz, 2 H) 3.30 - 3.39 (m, 2 H) 2.77 - 2.87 (m, 1 H) 2.47 (br t, 7 =6.80 Hz, 2 H) 1.95 - 2.03 (m, 1 H) 1.63 - 1.76 (m, 3 H)

LCMS (ESI+) : m/z 469.0 (M+H)

compound 221:

J H NMR (400 MHz, DMSO-de) d ppm 10.50 - 10.34 (m, 1H), 7.60 - 7.50 (m, 1H), 7.44 - 7.38

(m, 2H), 7.36 - 7.25 (m, 2H), 6.86 - 6.82 (m, 1H), 3.90 - 3.80 (m, 2H), 3.80 - 3.74 (m, 2H), 3.59

- 3.52 (m, 2H), 3.32 - 3.25 (m, 2H), 2.76 - 2.68 (m, 2H), 2.45 - 2.38 (m, 2H), 2.35 - 2.31 (m, 1H), 2.29 - 2.25 (m, 3H), 1.98 - 1.86 (m, 1H), 1.67 - 1.62 (m, 2H)

LCMS (ESI+): m/z 449.1 (M+H)

compound 222: J H NMR (400 MHz, DMSO -d 6 ) d ppm 10.51 - 10.46 (m, 1H), 7.58 - 7.54 (m, 1H), 7.36 - 7.31 (m, 1H), 7.30 - 7.28 (m, 1H), 7.27 - 7.25 (m, 1H), 7.21 - 7.17 (m, 1H), 6.62 - 6.59 (m, 1H), 3.90 - 3.81 (m, 2H), 3.80 - 3.76 (m, 2H), 3.75 - 3.72 (m, 3H), 3.56 (s, 2H), 3.37 (br s, 2H), 2.79 - 2.56 (m, 2H), 2.39 (br s, 2H), 1.99 - 1.86 (m, 1H), 1.67 - 1.63 (m, 2H), 0.07 - 0.06 (m, 1H)

LCMS (ESI+) : m/z 465.1 (M+H)

compound 220

J H NMR (400MHz, DMSO-de) d ppm 10.81 (s, 1 H), 7.74 (t, / = 1.8 Hz, 1 H), 7.53 (d, / = 2.7 Hz, 1 H), 7.49 (s, 1 H), 7.32 (d, / = 2.7 Hz, 1 H), 7.12 (t, / = 1.5 Hz, 1 H), 4.04 - 3.89 (m, 2 H), 3.85 - 3.73 (m, 3 H), 3.66 - 3.53 (m, 3 H), 3.43 - 3.37 (m, 1 H), 2.43 (quin, / = 6.9 Hz, 2 H), 2.37 - 2.27 (m, 1 H), 1.88 (qd, / = 7.8, 12.3 Hz, 1 H).

LCMS (ESI+): m/z 491.0 (M+H)

EXAMPLE 28: Synthesis of compound 229:

Experimental Procedures:

Preparation of compound 2:

To a solution of compound 1 (25 g, 143.2 mmol, 1 eq) in 200 mL of dichloromethane was added (COCl)2 (72.7 g, 572.9 mmol, 50.2 mL, 4 eq) and DMF (1.0 g, 14.3 mmol, 1.1 mL, 0.1 eq). The mixture was stirred at 20°C for 1 hour. After reaction, it was concentrated under reduced pressure, then it was quenched by addition of 30 mL of methanol. The resulting solution was concentrated under reduced pressure to give 27 g of crude compound 2 as a yellow oil, which was used directly into the next step without purification.

Preparation of compound 3:

To a solution of compound 2 (27 g, 143.2 mmol, 1 eq ) in 250 mL of H2SO4 (98% purity) was added HNO3 (14.6 g, 157.5 mmol, 10.4 mL, 68% purity, 1.1 eq) at 0°C. The mixture was stirred at 0°C for 1 hour. After reaction, it was added to 150 mL of ice water and stirred at 0°C for 0.2 hour. Then it was partitioned between 100 mL of water and 200 mL of dichloromethane . The organic phase was separated, washed with 150 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to get 33 g of crude compound 3 as a yellow oil, which was used directly into the next step without purification.

Preparation of compound 4:

To a solution of compound 3 (63 g, 269.7 mmol, 1 eq) in 600 mL of ethyl alcohol was added NH4CI (72.1 g, 1.4 mol, 5 eq) and 120 mL of H2O. The mixture was warmed to 80°C, then it was added Fe (45.2 g, 809.1 mmol, 3 eq) slowly. The mixture was stirred at 80°C for 1 hour. Then it was filtered and the filtrate was concentrated under reduced pressure, the residue was partitioned between 250 mL of water and 750 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate = 1:0 to 4:1) to get 21 g of compound 4 (103.1 mmol, 38.2% yield) as a yellow solid.

Preparation of compound 5:

4 5

To a solution of compound 4 (15 g, 73.7 mmol, 1 eq ) in 150 mL of dichloromethane was added TEA (11.2 g, 110.5 mmol, 15.4 mL, 1.5 eq) and 3-chloropropane-l-sulfonyl chloride (19.6 g, 110.5 mmol, 13.4 mL, 1.5 eq) at 0°C. The mixture was stirred at 20°C for 24 hours. After reaction, it was concentrated under reduced pressure to give a residue, which was added 150 mL of methanol and TEA (11.2 g, 110.5 mmol, 15.4 mL, 1.5 eq). The mixture was stirred at

65°C for 12 hours and 70°C for 3 hours. Then it was concentrated under reduced pressure, the residue was partitioned between 200 mL of water and 500 mL of ethyl acetate. The organic phase was separated, washed with 100 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column

chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =1:0 to 0:1).

The crude product was then re-crystallized from 50 mL of ethyl acetate at 60°C to get 11.8 g of compound 5 (38.2 mmol, 51.8% yield, 99.5% purity) as a light yellow solid.

Preparation of compound 229:

To a solution of methyl compound 5 (50 mg, 162.5 pmol, 1 eq) in 1 mL of toluene was added 3-chloro-5-(trifluoromethoxy)aniline (37.8 mg, 178.7 pmol, 1.1 eq) and AlMe3 (2 M, 81.2 pL, 1 eq) at 0°C. The mixture was stirred at 100°C for 12 hours. Then it was added 2 mL of methanol and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (TFA condition) to get 23.2 mg of 229 (47.5 pmol, 29.2% yield, 99.7% purity) as a pink solid.

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 8.16 - 8.23 (m, 1 H) 7.60 - 7.66 (m, 1 H) 7.55 (s, 1 H) 7.28 - 7.32 (m, 2 H) 7.05 - 7.09 (m, 1 H) 3.74 - 3.83 (m, 2 H) 3.39 - 3.48 (m, 2 H) 2.59 (quin, J = 7.00 Hz, 2 H)

LCMS (ESI+): m/z 486.9 (M+H)

EXAMPLE 29: Synthesis of compound 230:

Experimental Procedures:

Preparation of compound 5:

5 6

To a solution of compound 5 (11.8 g, 38.2 mmol, 99.5% purity, 1 eq ) in 100 mL of methanol was added NaOH (4.6 g, 114.5 mmol, 3 eq) and 20 mL of H2O. The mixture was stirred at 20°C for 12 hours. Then it was concentrated under reduced pressure, the residue was partitioned between 50 mL of water and 250 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to get 9.9 g of crude compound 6 as a white solid, which was used directly into the next step without purification.

Preparation of 230:

To a soludon of methyl 3-amino-5-chloro-benzoate (31.6 mg, 170.2 pmol, 1 eq) and compound 6 (50 mg, 170.2 pmol, 1 eq) in 1 mL of DMF was added TEA (51.7 mg, 510.7 pmol, 3 eq) and HATU (77.7 mg, 204.3 pmol, 1.2 eq) at 0°C. The mixture was stirred at 20°C for 12 hours. Then it was purified by prep- HPLC (TFA condition) to get 14.1 mg of 230 (30.1 pmol, 17.7% yield, 98.5% purity) as a white solid.

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 8.14 - 8.17 (m, 1 H) 8.10 - 8.14 (m, 1 H) 7.97 - 8.00 (m, 1 H) 7.83 - 7.86 (m, 1 H) 7.32 - 7.37 (m, 2 H) 3.93 - 3.96 (m, 3 H) 3.79 - 3.83 (m, 2 H) 3.42 - 3.47 (m, 2 H) 2.56 - 2.64 (m, 2 H)

LCMS (ESI+) : m/z 461.0 (M+H)

The following compounds were prepared analogously coupling the appropriate aniline with the acid 6:

F

compound 261:

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 7.86 - 7.96 (m, 1 H) 7.30 - 7.35 (m, 1 H) 7.20 - 7.25 (m, 2 H) 6.71 - 6.78 (m, 1 H) 3.82 - 3.86 (m, 3 H) 3.76 - 3.81 (m, 2 H) 3.40 - 3.46 (m, 2 H)

2.54 - 2.63 (m, 2 H)

LCMS (ESI+): m/z 433.0 (M+H)

F

compound 264

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 7.79 - 7.87 (m, 1 H) 7.33 (dd, / = 10.38, 2.13 Hz, 1 H) 7.25 - 7.27 (m, 1 H) 6.85 - 6.89 (m, 2 H) 6.30 - 6.34 (m, 1 H) 3.81 - 3.86 (m, 6 H) 3.75 - 3.80 (m, 2 H) 3.38 - 3.46 (m, 2 H) 2.54 - 2.63 (m, 2 H) LCMS (ESI+): m/z 429.0 (M+H)

EXAMPLE 30: Synthesis of 246

The amine of 246 was made through a Suzuki reaction.

Preparation of compound 2:

To a solution of compound 1 (5 g, 24.2 mmol, 1 eq), 4,4,5,5-tetramethyl-2- (4, 4, 5, 5- tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (6.8 g, 26.6 mmol, 1.1 eq) and KOAc (4.8 g, 48.4 mmol, 2 eq) in 100 mL of dioxane was added Pd(dppf)Cl2 (1.8 g, 2.4 mmol, 0.1 eq) under N2. The mixture was stirred at 110°C for 5 hours. Then it was partitioned between 100 mL of water and 150 mL of ethyl acetate. The organic phase was separated, washed with 100 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product. It was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate = 20:1 to 3:1) to get 5 g of compound 2 (19.7 mmol, 81.4% yield) as a yellow gum.

Preparation of compound 3:

To a solution of compound 2 (300 mg, 1.2 mmol, 1 eq ), 2-bromopyrimidine (206.9 mg, 1.3 mmol, 1.1 eq) and K2CO3 (490.6 mg, 3.6 mmol, 3 eq) in 0.8 mL of H2O and 4 mL of dioxane was added Pd(dppf)Cl2 (86.6 mg, 118.3 pmol, 0.1 eq) and purged with N2 for 3 times. The mixture was stirred at 100°C for 12 hours. Then it was filtered, partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =1 :0 to 1 :1) to get 100 mg of compound 3 (486.3 pmol, 41.1% yield) as a white solid.

Then compound 3 was used into an amide coupling reaction through a normal method to give

246.

F

compound 246:

J H NMR (400 MHz, DMSO-de) d ppm 10.98 - 11.07 (m, 1 H) 8.88 - 9.02 (m, 2 H) 8.68 - 8.77 (m, 1 H) 8.11 - 8.20 (m, 1 H) 7.96 - 8.06 (m, 1 H) 7.46 - 7.57 (m, 1 H) 7.30 - 7.40 (m, 1 H) 7.25 - 7.29 (m, 1 H) 3.74 - 3.88 (m, 2 H) 3.55 - 3.70 (m, 2 H) 2.40 - 2.46 (m, 2 H)

LCMS (ESI+): m/z 481.0 (M+H)

The amine of 242 was made by using below method:

Experimental Procedures:

Preparation of compound 2:

1 2 To a solution of l-chloro-3-methoxy-5-nitro-benzene (5 g, 26.7 mmol, 1 eq) in 30 mL of dichloromethane was added BBr3 (20.0 g, 80.0 mmol, 7.7 mL, 3 eq) at 0°C. The mixture was stirred at 15°C for 12 hours. Then it was partitioned between 30 mL of water and 50 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =100:1 to 30: 1) to give 4.3 g of compound 2 (4.3 g, 24.8 mmol, 93.0% yield) as a white solid.

Preparation of compound 3:

2 3

A mixture of 3-chloro-5-nitro-phenol (1 g, 5.8 mmol, 1 eq), Fe (1.6 g, 28.8 mmol, 5 eq) and NH4CI (3.1 g, 57.6 mmol, 10 eq) in 10 mL of MeOH and 4 mL of water was degassed and purged with N2 for 3 times, and then the mixture was stirred at 70°C for 12 hours under N2 atmosphere. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. Then it was partitioned between 2 mL of water and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na 2 S0 4 , filtered and concentrated under reduced pressure to give 1 g of crude compound 3 as a yellow oil, which was used directly into the next step.

Preparation of compound 4:

To a solution of 3-amino-5-chloro-phenol (440 mg, 3.1 mmol, 1 eq) and KOH (1.7 g, 30.7 mmol, 10 eq) in 5 mL of acetonitrile and 5 mL of water was added dropwise 1- [[bromo(difluoro)methyl]-ethoxy-phosphoryl]oxye thane (982.0 mg, 3.7 mmol, 1.2 eq) at -78°C. After addition, the resulting mixture was stirred at 20°C for 12 hours. Then it was partitioned between 2 mL of water and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, Petroleum ether : Ethyl acetate = 2: 1) to give 80 mg of compound 4 (413.3 pmol, 13.5% yield) as a yellow oil.

Then compound 4 was used into an amide coupling reaction by using a normal method to give 242.

J H NMR (400 MHz, DMSO -d 6 ) d ppm 11.01 (s, 1 H) 7.69 (t, 7 = 1.65 Hz, 1 H) 7.53 (s, 1 H)

7.49 (s, 1 H) 7.36 (dd, 7 = 11.25, 2.57 Hz, 1 H) 7.31 (s, 1 H) 7.25 (d, 7 = 1.34 Hz, 1 H) 7.12 (d, 7 = 2.32 Hz, 1 H) 7.11 (br s, 1 H) 3.81 (t, 7 = 6.54 Hz, 2 H) 3.62 (t, 7 = 7.34 Hz, 2 H) 2.44 (t, 7 = 6.97 Hz, 2 H)

LCMS (ESI+): m/z 469.0 (M+H)

EXAMPLE 31: Synthesis of compound 265

The amine of 265 was made through a Simmons-Smith reaction.

Experimental Procedures:

Preparation of compound 1:

1A 1 To a solution of 3-bromo-5-chloro-aniline (10 g, 48.4 mmol, 1 eq) and tert- butoxycarbonyl tert-butyl carbonate (12.7 g, 58.1 mmol, 13.4 mL, 1.2 eq) in 100 mL of THF was added K2CO3 (20.1 g, 145.3 mmol, 3 eq). The mixture was stirred at 40°C for 12 hrs. The reaction mixture was concentrated under reduced pressure to remove THF. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether/ethyl acetate=l/0 to 0/1) to give 6.1 g of compound 1 (19.90 mmol, 41.1% yield) as a white solid.

Preparation of compound 2:

To a solution of compound 1 (2 g, 6.5 mmol, 1 eq), 2-isopropenyl-4,4,5,5- tetramethyl- 1,3,2-dioxaborolane (1.3 g, 7.8 mmol, 1.2 eq) and K2CO3 (1.8 g, 13.1mmol, 2 eq) in 30 mL of dioxane and 6 mL of H2O was added Pd(dppf)Cl2 (477.3 mg, 652.4 pmol, 0.1 eq) . The mixture was stirred at 110°C for 12 hours. Then it was partitioned between 30 mL of water and 60 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column

chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate = 20:1 to 7:1) to give 1.3 g of compound 2 (1.3 g, 4.8 mmol, 74.4% yield) as a white solid.

Preparation of compound 3:

To a solution of ZnEt2 (1 M, 18.7 mL, 5 eq.) in 100 mL of dichloromethane was added CH2I2 (5.0 g, 18.7 mmol, 1.5 mL, 5 eq.) dropwise at 0°C. The mixture was stirred at 0°C for 0.3 hour. TFA (2.1 g, 18.7 mmol, 1.4 mL, 5 eq.) was added to the mixture dropwise at 0°C. The mixture was stirred at 0°C for 0.3 hour. Then a solution of compound 2 (1 g, 3.7 mmol, 1 eq) in 50 mL of dichloromethane was added to the mixture dropwise at 0°C. Then the mixture was warmed to 20°C and stirred for 3 hours. After reaction, it was diluted with 50 mL of water and 50 mL of ethyl acetate, separated and the organic layer and washed with 30 mL of brine. The collected organic layer was dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to get a residue, which was purified by column

chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate = 50: 1 to 15: 1) to give 480 mg of compound 3 (1.6 mmol, 43.5% yield, TFA salt) as a brown oil.

Then compound 3 was used into an amide coupling reaction by using a normal method to give

265.

p

J H NMR (400 MHz, DMSO-d6) d ppm 10.79 - 10.72 (m, 1H), 7.72 - 7.66 (m, 1H), 7.56 - 7.50 (m, 1H), 7.48 - 7.42 (m, 1H), 7.35 - 7.29 (m, 1H), 7.03 - 6.98 (m, 1H), 3.86 - 3.75 (m, 2H), 3.66 - 3.56 (m, 2H), 2.45 - 2.40 (m, 2H), 1.39 - 1.34 (m, 3H), 0.88 - 0.82 (m, 2H), 0.82 - 0.76 (m, 2H)

LCMS (ESI+): m/z 475.0 (M+H)

EXAMPLE 32: Synthesis of 292, 238

The amine of 292, 238 was made by using below method:

6

Experimental procedure:

The preparation of compound 4 can be found in report of 224, 225.

Preparation of compound 2:

To a solution of compound 1 (1 g, 5.1 mmol, 1 eq ), compound 2 (1.4 g, 5.6 mmol, 1.1 eq .) and KOAc (996.2 mg, 10.2 mmol, 2 eq) in 30 mL of dioxane was added Pd(dppf)Cl2 (371.4 mg, 507.5 pmol, 0.1 eq) under N2. The mixture was stirred at 110°C for 12 hours. Then it was partitioned between 100 mL of water and 90 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of brine, dried over Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate = 20: 1 to 3: 1) to give 820 mg compound 3 (3.4 mmol, 66.2% yield) as a yellow gum. Preparation of compound 5:

To a solution of compound 3 (800 mg, 3.3 mmol, 1 eq ), compound 4 (3.6 g, 16.4 mmol,

5 eq) and CS2CO3 (2.1 g, 6.6 mmol, 2 eq) in 10 mL of dioxane and 2 mL of H2O was added Pd(dppf)Cl2 (239.8 mg, 328.0 pmol, 0.1 eq) under N2. The mixture was stirred at 100°C for 12 hours. Then it was partitioned between 30 mL of water and 30 mL of ethyl acetate. The organic phase was separated, washed with 30 mL of brine, dried over Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate = 20: 1 to 1 : 1) to 420 mg of compound 5 (2.3 mmol, 68.8% yield) as a yellow solid.

Preparation of compound 6

5 6

To a solution of 3-amino-5-(2,5-dihydrofuran-3-yl)benzonitrile (420 mg, 2.3 mmol, 1 eq) in 50 mL of ethyl acetate was added Pd/C (100 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with ¾ several times. The mixture was stirred at 20°C for 1 hour under ¾ (15 psi) atmosphere. After reaction, it was filtered and concentrated under reduced pressure to give crude 420 mg of compound 6 as a light yellow oil.

Then compound 6 was used into an amide coupling reaction by using a normal method to give 292.

Compound 292: J H NMR (400 MHz, DMSO -d 6 ) d ppm 11.00 - 10.97 (m, 1H), 8.06 - 8.00 (m, 1H), 7.87 - 7.81 (m, 1H), 7.57 - 7.53 (m, 1H), 7.38 - 7.32 (m, 1H), 7.26 - 7.21 (m, 1H), 4.04 - 3.98 (m, 1H), 3.97 - 3.93 (m, 1H), 3.83 - 3.76 (m, 3H), 3.64 - 3.55 (m, 3H), 3.51 - 3.41 (m, 1H), 2.47 - 2.41 (m, 2H), 2.40 - 2.29 (m, 1H), 1.96 - 1.85 (m, 1H)

LCMS (ESI+) : m/z 464.1 (M+H)

238 was prepared analogously coupling the appropriate acid with the aniline 6:

J H NMR (400 MHz, DMSO-de) d ppm 10.92 - 10.86 (m, 1H), 8.07 - 8.01 (m, 1H), 7.89 - 7.84 (m, 1H), 7.62 - 7.56 (m, 1H), 7.54 - 7.51 (m, 1H), 7.40 - 7.31 (m, 2H), 4.04 - 4.00 (m, 1H), 3.94 (br s, 1H), 3.82 (s, 3H), 3.62 - 3.53 (m, 3H), 3.51 - 3.39 (m, 1H), 2.53 - 2.52 (m, 2H), 2.45 - 2.40 (m, 2H), 2.40 - 2.34 (m, 1H), 1.96 - 1.85 (m, 1H)

LCMS (ESI+): m/z 446.0 (M+H)

EXAMPLE 33: Synthesis of compound 235:

Experimental Procedures:

Preparation of compound 2:

A mixture of 3-bromo-5-chloro-aniline (20 g, 96.8 mmol, 1 eq), 2-(3, 6-dihydro- 2H- pyran-4-yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (20.3 g, 96.8 mmol, 1 eq), K2CO3 (40.2 g, 290.6 mmol, 3 eq), Pd(dppf)Cl2 (7.1 g, 9.7 mmol, 0.1 eq) in 200 mL of dioxane and 20 mL of water was degassed and purged with N2 for 3 times, and then the mixture was stirred at 110 °C for 12 hours under N2 atmosphere. After reaction, it was partitioned between 500 mL of water and 100 mL of ethyl acetate. The organic phase was separated, washed with 200 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =20: 1 to 5: 1) to give 20 g of compound 2 as a light yellow solid.

Preparation of compound 3:

To a solution of 3-chloro-5-(3,6-dihydro-2H-pyran-4-yl)aniline (6 g, 28.6 mmol, 1 eq) in 30 mL of ethyl acetate was added Pd/C (500 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with ¾ for three times. The mixture was stirred at 20°C for 12 hours under ¾ (15psi) atmosphere. After reaction, the reaction mixture was filtered, and concentrated under reduced pressure to give 16.2 g of compound 3 as a yellow solid, which was used into the next step without further purification. Preparation of compound 235:

To a solution of 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)-3-fluoro-benzoic acid (5 g, 17.0 mmol, 1 eq ), TEA (3.5 g, 34.1 mmol, 4.7 mL, 2 eq ) and 3-chloro-5-tetrahydropyran- 4-yl- aniline (3.6 g, 17.0 mmol, 1 eq) in 60 mL of dimethyl formamide was added dropwise HATU (7.1 g, 18.7 mmol, 1.1 eq) in 30 mL of dimethyl formamide at 0°C. After addition, the resulting mixture was stirred at 20°C for 1 hour. Then the mixture was stirred at 30°C for 12 hours. After reaction, it was added 200 mL of water dropwise, the mixture was filtered and concentrated under reduced pressure to give the crude product, which was purified by recrystallization in dichloromethane and methanol. The crude product was added into 150 mL of dichloromethane at 40°C, 2 mL of methnol was added dropwise to the mixture with stirring for 0.5 hour to give a light yellow solution. Then the mixture was cooled naturally to 20°C over 1 hour. The mixture was filtered and the filter cake was dried in vacuum at 40°C to get 3.7g of 235 (3.7 g, 7.6 mmol, 44.8% yield, 99.0% purity) as a white solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.79 (s, 1 H) 7.71 (t, 7 = 1.77 Hz, 1 H) 7.51 (s, 1 H) 7.35 (dd, 7 = 11.13, 2.57 Hz, 1 H) 7.19 - 7.25 (m, 1 H) 7.12 (s, 1 H) 3.95 (br dd, 7=10.94, 3.36 Hz, 2 H) 3.81 (t, 7 = 6.54 Hz, 2 H) 3.62 (t, 7 = 7.34 Hz, 2 H) 3.43 (td, 7 = 11.55, 1.83 Hz, 2 H) 2.75 - 2.85 (m, 1 H) 2.39 - 2.46 (m, 2 H) 1.56 - 1.76 (m, 4 H)

LCMS (ESI+): m/z 487.0 (M+H)

EXAMPLE 34: Synthesis of compound 241:

Experimental Procedures:

Preparation of compound 7:

To a solution of compound 6 (2 g, 6.8 mmol, 1 eq ), TEA (1.4 g, 13.6 mmol, 1.9 mL, 2 eq ) and 3-bromo-5-chloro-aniline (1.4 g, 6.8 mmol, 1 eq) in 20 mL of DMF was cooled to 0°C, HATU (2.7 g, 7.2 mmol, 1.1 eq) in 15 mL of DMF was added dropwise. After addition, the resulting mixture was stirred at 20°C for 12 hours. Then it was partitioned between 50 mL of water and 150 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =10:1 to 0:1) to get 2.9 g of compound 7 (6.0 mmol, 88.3% yield) as a red solid. Preparation of 241:

7 241

To a solution of compound 7 (60 mg, 124.4 m ihoI, 1 eq), 4-pyridylboronic acid (16.8 mg, 136.89 pmol, 1.1 eq) and K2CO3 (51.6 mg, 373.3 pmol, 3 eq) in 1 mL of dioxane and 0.2 mL of H2O was added Pd(dppf)Cl2 (9.1 mg, 12.4 pmol, 0.1 eq) and purged with N2 for 3 times. The mixture was stirred at 110°C for 12 hours. After reaction, it was filtered, the filtrate was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (neutral condition) to get 27.9 mg of 241 (58.1 pmol, 46.7% yield, 100.0% purity) as a white solid.

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 8.67 - 8.74 (m, 2 H) 8.02 - 8.11 (m, 1 H) 7.83 - 7.90 (m, 1 H) 7.73 - 7.80 (m, 1 H) 7.48 - 7.53 (m, 2 H) 7.44 - 7.46 (m, 1 H) 7.32 - 7.37 (m, 2 H) 3.78 - 3.83 (m, 2 H) 3.42 - 3.47 (m, 2 H) 2.56 - 2.64 (m, 2 H)

LCMS (ESI+): m/z 480.0 (M+H)

The following compounds were prepared analogously to compound 241 using different borate to do a Suzuki Reaction:

compound 247:

J H NMR (400 MHz, DMSO-de) d ppm 11.03 - 10.98 (m, 1H), 8.99 - 8.92 (m, 1H), 8.73 - 8.66

(m, 1H), 8.27 - 8.19 (m, 1H), 7.98 - 7.89 (m, 2H), 7.70 - 7.61 (m, 2H), 7.39 - 7.33 (m, 1H), 7.30 - 7.25 (m, 1H), 3.84 - 3.79 (m, 2H), 3.65 - 3.59 (m, 2H), 2.45 - 2.41 (m, 2H)

LCMS (ESI+): m/z 480.0 (M+H) compound 248:

J H NMR (400 MHz, DMSO-de) d ppm 11.03 (s, 1 H) 9.24 (s, 1 H) 9.12 (s, 2 H) 7.94 (br d, / = 12.35 Hz, 2 H) 7.71 (s, 1 H) 7.36 (br dd, J = 11.19, 2.38 Hz, 1 H) 7.26 (s, 1 H) 3.81 (br t, / = 6.42 Hz, 2 H) 3.62 (br t, / = 7.34 Hz, 2 H) 2.42 - 2.46 (m, 2 H)

LCMS (ESI+) : m/z 481.0 (M+H)

EXAMPLE 35: Synthesis of 251 and 250

A mixture of compound ? (0.1 g, 207.4 pmol, 1 eq), tributyl(2-pyridyl)stannane (91.6 mg, 248.9 pmol, 1.2 eq), Pd(PPli3)2Cl2 (14.6 mg, 20.7 pmol, 0.1 eq) in 1 mL of Tol. was degassed and purged with N2 for 3 times, and then the mixture was stirred at 100°C for 12 hours under N2 atmosphere. The reaction mixture was filtered and the filtrate was concentrated in vacuum, which was purified by prep- HPLC (basic condition) to give 10.5 mg of Compound 251 (21.9 pmol, 10.5% yield, 100.0% purity) as a white solid.

JH NMR (400 MHz, DMSO-de) d ppm 10.98 (s, 1 H) 8.70 (d, J = 4.65 Hz, 1 H) 8.38 (s, 1 H) 7.96 - 8.01 (m, 1 H) 7.88 - 7.96 (m, 3 H) 7.40 - 7.45 (m, 1 H) 7.35 (dd, / = 11.13, 2.57 Hz, 1 H) 7.27 (s, 1 H) 3.82 (t, J = 6.48 Hz, 2 H) 3.62 (t, J = 7.34 Hz, 2 H) 2.40 - 2.46 (m, 2 H)

LCMS (ESI+): m/z 480.0 (M+H) Compound 250 was made through a Buchwald reaction:

7 250

A mixture of compound 7 (90 mg, 186.7 pmol, 1 eq), morpholine (19.5 mg, 224.0 pmol, 19.71 pL, 1.2 eq), t-BuOK (31.4 mg, 280.0 pmol, 1.5 eq), Xantphos (21.6 mg, 37.3 pmol, 0.2 eq) and Pd2(dba)3 (34.2 mg, 37.3 pmol, 0.2 eq) in 2 mL of dioxane was degassed and purged with N2 for 3 times, and then the mixture was stirred at 110 °C for 12 hours under N2 atmosphere. After reaction, it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =0: 1) firstly. The obtained crude product was re-purified by prep- HPLC (TFA condition) to get 3.5 mg of 250 (5.2 pmol, 2.8% yield, 89.4% purity, TFA salt) as a white solid.

J H NMR (400MHz, CHLOROFORM-d) d ppm 7.94 (s, 1H), 7.34 - 7.28 (m, 2H), 7.09 (s, 1H), 6.73 (s, 1H), 3.91 - 3.84 (m, 4H), 3.78 (t, / = 6.5 Hz, 2H), 3.43 (t, / = 7.5 Hz, 2H), 3.25 - 3.19 (m, 4H), 2.58 (quin, J = 7.0 Hz, 2H)

LCMS (ESI+): m/z 488.0 (M+H)

Compounds 215, 305 were made analogously to 250 by using the appropriate aryl bromide and corresponding amine to do a Buchwald Reaction:

compound 215:

J H NMR (400 MHz, DMSO-de) d ppm 10.66 (s, 1 H), 7.51 (d, / = 2.63 Hz, 1 H), 7.29 (s, 2 H), 7.18 (s, 1 H), 6.90 (s, 1 H), 3.74 - 3.82 (m, 6 H), 3.59 (t, / = 7.45 Hz, 2 H), 3.13 (br s, 4 H), 2.41 (br t, / = 7.02 Hz, 2 H).

LCMS (ESI+): m/z 554.0 (M+H)

Experimental Procedures:

Preparation of compound 8:

To a solution of compound 7 (1 g, 2.1 mmol, 1 eq), 4,4,5,5-tetramethyl-2- (4, 4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (579.4 mg, 2.3 mmol, 1.1 eq) and

AcOK (407.1 mg, 4.2 mmol, 2 eq) in 15 mL of dioxane was added Pd(dppf)Cl2 (151.8 mg, 207.4 pmol, 0.1 eq) and purged with N2 for 3 times. The mixture was stirred at 100°C for 12 hours. Then it was filtered, the filtrate was partitioned between 50 mL of water and 250 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate = 5:1 to 1 :1) to get 1.0 g of compound 8 (2.0 mmol, 95.7% yield) as a white solid.

Preparation of 246:

To a solution of compound 8 (100 mg, 189.0 pmol, 1 eq), 2-bromopyrimidine (30.0 mg, 189.0 pmol, 1 eq) and K2CO3 (78.4 mg, 566.9 pmol, 3 eq) in 1.5 mL of dioxane and 0.3 mL of H2O was added Pd(dppf)Cl2 (13.8 mg, 18.9 pmol, 0.1 eq) and purged with N2 for 3 times. The mixture was stirred at 110°C for 12 hours. Then it was filtered, the filtrate was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (TFA condition) to get 12.2 mg of 246 (23.4 pmol, 12.4% yield, 92.4% purity) as a brown solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.97 - 11.09 (m, 1 H) 8.92 - 9.01 (m, 2 H) 8.64 - 8.78 (m, 1 H) 8.12 - 8.19 (m, 1 H) 7.98 - 8.05 (m, 1 H) 7.48 - 7.56 (m, 1 H) 7.31 - 7.42 (m, 1 H) 7.26

- 7.29 (m, 1 H) 3.79 - 3.85 (m, 2 H) 3.59 - 3.65 (m, 2 H) 2.42 - 2.46 (m, 2 H)

LCMS (ESI+): m/z 481.0 (M+H)

The following compounds were prepared analogously to compound 246 using different aryl bromide:

compound 276:

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 9.25 - 9.47 (m, 1 H) 8.05 - 8.11 (m, 1 H) 7.63 - 7.70 (m, 1 H) 7.35 - 7.40 (m, 1 H) 7.30 (br s, 1 H) 7.20 - 7.25 (m, 1 H) 7.18 (br s, 1 H) 6.63 - 6.74 (m, 1 H) 6.46 - 6.51 (m, 1 H) 3.69 - 3.78 (m, 2 H) 3.50 (s, 3 H) 3.33 (br s, 2 H) 2.46 - 2.56 (m, 2 H)

LCMS (ESI+) : m/z 510.0 (M+H)

compound 249:

J H NMR (400MHz, DMSO-de) d ppm 10.86 (s, 1 H), 7.85 - 7.73 (m, 2 H), 7.56 (br d, / = 8.6

Hz, 2 H), 7.42 (s, 1 H), 7.33 (br dd, / = 2.1, 11.1 Hz, 1 H), 7.23 (br s, 1 H), 7.25 - 7.20 (m, 1 H),

7.03 (br d, / = 8.6 Hz, 2 H), 4.17 - 4.04 (m, 2 H), 3.78 (br t, / = 6.4 Hz, 2 H), 3.68 - 3.62 (m, 2 H), 3.59 (br t, / = 7.4 Hz, 2 H), 3.29 (s, 3 H), 2.43 - 2.38 (m, 2 H)

LCMS (ESI+): m/z 553.0 (M+H)

The aryl bromide of 244 was made through a substitution reaction:

2

To a solution of 2-methoxyethanol (65.9 mg, 866.7 m ihoI, 1 eq) in 2 mL of DMF was added NaH (52.0 mg, 1.3 mmol, 60% purity, 1.5 eq) at 0°C under N2. The mixture was stirred at 0°C for 5 min. Then 5-bromo-2-chloro-pyridine (200 mg, 1.0 mmol, 1.2 eq) was added. The mixture was stirred at 20°C for 2 hours. After reaction, it was quenched by addition of 5 mL of saturated aqueous NH4CI and extracted with 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated in vacuum. The residue was purified by prep-TLC (Si02, eluting with petroleum ether:ethyl acetate = 10: 1) to give 85 mg of compound 2 (366.3 pmol, 42.3% yield) as a yellow oil.

Then compound 2 was used into a Suzuki reaction by using a normal method to give 244.

F

compound 244:

J H NMR (400 MHz, DMSO-de) d ppm 10.95 - 10.92 (m, 1H), 8.47 - 8.42 (m, 1H), 8.02 - 7.96 (m, 1H), 7.87 - 7.81 (m, 2H), 7.55 - 7.50 (m, 1H), 7.39 - 7.32 (m, 1H), 7.28 - 7.23 (m, 1H), 6.98

- 6.92 (m, 1H), 4.45 - 4.40 (m, 2H), 3.84 - 3.78 (m, 2H), 3.69 - 3.66 (m, 2H), 3.63 - 3.60 (m, 2H), 3.32 - 3.29 (m, 3H), 2.45 - 2.40 (m, 2H)

LCMS (ESI+): m/z 554.1 (M+H)

EXAMPLE 37: Synthesis of compound 287 and 288:

Experimental Procedures:

Preparation of compound 2A, 2B:

To a solution of 3-bromo-5-chloro-aniline (2 g, 9.7 mmol, 1 eq), 2,3-dihydrofuran (1.4 g, 19.4 mmol, 1.5 mL, 2 eq), PPI13 (508.3 mg, 1.9 mmol, 0.2 eq) and K2CO3 (4.0 g, 29.1 mmol, 3 eq) in 20 mL of DMF was added Pd(OAc)2 (217.6 mg, 969.0 pmol, 0.1 eq) at 20°C under N2. The mixture was stirred at 110°C for 12 hours. Then it was partitioned between 20 mL of water and 40 mL of ethyl acetate. The organic phase was separated, washed with 60 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, Petroleum ether : Ethyl acetate = 100: 1 to 10: 1) to give 780 mg of compound 2A (4.0 mmol, 41.1% yield) as a yellow oil and 470 mg of compound 2B (2.4 mmol, 24.8% yield) as a yellow oil.

Preparation of compound 3:

To a solution of 3-chloro-5-(2,3-dihydrofuran-5-yl)aniline (0.7 g, 3.5 mmol, 1 eq) in 1 mL of ethyl acetate was added Pd/C (0.2 g, 10% purity) under ¾ atmosphere. The suspension was degassed and purged with ¾ for 3 times. Then it was stirred at 20°C for 20 min under ¾ (15 Psi) atmosphere. After reaction, it was filtered and the filtrate was concentrated in vacuum, which was purified by prep-TLC (Si(¾, Petroleum ether : Ethyl acetate = 100: 1 to 5: 1) to give 480 mg of compound 3 (2.4 mmol, 69.9% yield) as a yellow oil.

To a solution of 3-chloro-5-tetrahydrofuran-2-yl-aniline (0.4 g, 2.2 mmol, 1 eq) and methyl 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)-3-fluoro-benzoat e (736.4 mg, 2.4 mmol, 1.1 eq) in 4 mL of Tol. was added dropwise AlMe3 (2 M, 1.6 mL, 1.5 eq) at 0°C. After addition, the mixture was stirred at 110°C for 12 hours. Then it was partitioned between 5 mL of water and 10 mL of ethyl acetate. The organic phase was separated, washed with 6 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was triturated with 5 mL of ethyl acetate at 20°C for 1 hour. The resulting mixture was filtered and the filter cake was dried in vacuum to give 420 mg of compound 5 (887.3 pmol, 40.8% yield) as a yellow solid.

Preparation of compound 287 and 288:

Compound 5 (420 mg, 887.3 m mol ) was further separated by SFC to give 120.8 mg of 287

(255.2 pmol, 28.8% yield, 100% purity) and 125.5 mg of 288 (256.4 pmol, 28.9% yield, 96.7 % purity). The structures of the isomers were random assignment.

287:

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 8.56 (s, 1 H) 7.72 (s, 1 H) 7.30 (s, 1 H) 7.19 (br d, / = 10.26 Hz, 1 H) 7.13 (br s, 1 H) 7.08 (s, 1 H) 4.72 (br t, / = 7.07 Hz, 1 H) 4.02 (q, J = 7.09 Hz, 1 H) 3.88 (q, J = 7.38 Hz, 1 H) 3.69 (br t, / = 6.44 Hz, 2 H) 3.36 (br t, / = 7.38 Hz, 2 H) 2.52 (quin, J = 6.88 Hz, 2 H) 2.25 - 2.35 (m, 1 H) 1.91 - 2.02 (m, 2 H) 1.69 - 1.79 (m, 1 H)

LCMS (ESI+): m/z 473.0 (M+H)

288:

1H NMR (400 MHz, CHLOROFORM-c/) d ppm 8.49 (s, 1 H) 7.72 (s, 1 H) 7.31 (s, 1 H) 7.22 (dd, J = 10.26, 2.13 Hz, 1 H) 7.14 (br s, 1 H) 7.09 (s, 1 H) 4.74 (t, J = 7.07 Hz, 1 H) 4.03 (q, J = 7.05 Hz, 1 H) 3.85 - 3.94 (m, 1 H) 3.70 (br t, / = 6.44 Hz, 2 H) 3.37 (t, J = 7.38 Hz, 2 H) 2.53 (quin, J = 6.91 Hz, 2 H) 2.26 - 2.36 (m, 1 H) 1.93 - 2.02 (m, 2 H) 1.70 - 1.80 (m, 1 H)

LCMS (ESI+): m/z 473.0 (M+H)

EXAMPLE 38: Synthesis of compound 240:

Experimental Procedures:

Preparation of compound 2:

A mixture of compound 1 (5 g, 18.5 mmol, 1 eq), tributyl(l-ethoxyvinyl)stannane (7.4 g, 20.3 mmol, 6.9 mL, 1.1 eq), Pd(PPh3)2Ch (1.3 g, 1.8 mmol, 0.1 eq) in 100 mL of toluene was degassed and purged with N2 for 3 times, and then the mixture was stirred at 100°C for 12 hours under N2 atmosphere. Then the temperature was cooled to 25°C, 10 mL of 10% potassium fluoride aqueous solution was added and stirred for 2 hour, filtered, the filter cake was washed several times with ethyl acetate and the filtrate was evaporated to dryness. The residue was added 10 mL of IN HCI and stirred at 25°C for 2 hours. Then it was adjusted pH to 8 by using saturated sodium carbonate aqueous solution. Then it was partitioned between 100 mL of water and 300 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate = 100:1 to 50: 1) to give 3 g of compound 2 (12.9 mmol, 69.5% yield) as a colorless oil.

Preparation of compound 3:

A mixture of compound 2 (1.5 g, 6.4 mmol, 1 eq ), TEA (1.9 g, 19.3 mmol, 2.7 mL, 3 eq ) and Pd(dppf)Ch (470.1 mg, 642.4 pmol, 0.1 eq) in 20 mL of methanol was degassed and purged with CO for 3 times, and then the mixture was stirred at 65 °C for 12 hours under CO atmosphere (50 psi). Then it was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na 2 S0 4 , filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate = 30: 1 to 15:1) to give 1.1 g of compound 3 (5.2 mmol, 80.5% yield) as a white solid.

Preparation of compound 4:

To a solvent of 6 mL of BAST was added methyl 3-acetyl-5-chloro-benzoate (400 mg, 1.9 mmol, 1 eq). The mixture was stirred at 70°C for 12 hours. Then it was partitioned between 10 mL of icy saturated NaHCCb and 50 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and conentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate = 10: 1 to 5: 1) to give 300 mg of compound 4 (1.3 mmol, 67.9% yield) as a yellow oil. Preparation of compound 5:

To a solution of methyl 3-chloro-5-(l,l-difluoroethyl)benzoate (1.6 g, 6.8 mmol, 1 eq) in 16 mL of methanol was added NaOH (818.3 mg, 20.5 mmol, 3 eq) and 3.5 mL of water. The mixture was stirred at 20°C for 12 hours. Then it was partitioned between 50 mL of sat.NPLCl and 150 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give give 1.5 g of crude compound 5 as a yellow solid, which was used directly into the next step.

Preparation of compound 6:

To a solution of 3-chloro-5-(l,l-difluoroethyl)benzoic acid (300 mg, 1.4 mmol, 1 eq) in 3 mL of toluene was added dropwise DPPA (411.7 mg, 1.5 mmol, 1.1 eq) and TEA (151.4 mg, 1.5 mmol, 1.1 eq) at 25 °C. After addition, the mixture was stirred at this temperature for 1 hour, then warmed to 80°C and stirred for another 1 hour. 2-methylpropan-2-ol (151.2 mg, 2.0 mmol, 1.5 eq) was added dropwise. The resulting mixture was stirred at 110°C for 10 hours. After reaction, it was partitioned between 10 mL of saturated NPLCl and 50 mL of ethyl acetate. The organic phase was separated, washed with 25 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =30: 1 to 5:1) to give 200 mg of compound 6 (685.6 pmol, 50.4% yield) as a yellow solid. Preparation of compound 7:

6 7

To a solution of tert-butyl N-[3-chloro-5-(l, l-difluoroethyl)phenyl]carbamate (100 mg, 342.8 pmol, 1 eq) in 1 mL of dichloromethane was added TFA (462.0 mg, 4.1 mmol, 0.3 mL, 11.8 eq). The mixture was stirred at 25 °C for 1 hour. Then it was concentrated under reduced pressure to give a residue, which was added 2 mL of methanol and adjusted pH to 8 by using Na2C(¾ solid. The reaction mixture was filtered and concentrated under reduced pressure to give 60 mg of crude compound 7 as a yellow oil, which was used directly into the next step.

Preparation of compound 240:

7 F 240

To a solution of 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)-3-fluoro-benzoic acid (50 mg, 170.2 pmol, 1 eq) in 2 mL of dimethyl form amide was added compound 7 (32.6 mg, 170.2 pmol, 1 eq) and TEA (20.7 mg, 204.3 pmol, 1.2 eq). The mixture was cooled to 0°C, HATU (77.7 mg, 204.3 pmol, 1.2 eq) was added and stirred at 20°C for 12 hours. Then it was filtered and concentrated under reduced pressure to give a residue, which was purified by prep- HPLC (TFA condition) to give 11.4 mg of 240 (24.4 pmol, 14.3% yield, 100.0% purity) as a white solid.

J H NMR (400MHz, CHLOROFORM-d) d ppm 8.06 (br s, 1H), 7.87 (s, 1H), 7.61 (s, 1H), 7.35 - 7.28 (m, 3H), 3.79 (t, / = 6.5 Hz, 2H), 3.43 (t, / = 7.4 Hz, 2H), 2.59 (quin, J = 7.0 Hz, 2H), 2.02

- 1.87 (m, 3H)

LCMS (ESI+): m/z 467.0 (M+H) EXAMPLE 39: Synthesis of compound 245 and 253:

Experimental Procedures:

Preparation of compound 2:

To a solution of l,3-dibromo-5-chloro-benzene (1 g, 3.7 mmol, 1 eq) in 20 mL of terahydrofuran was added dropwise butyllithium (2.5 M, 1.6 mL, 1.1 eq) at -78°C under N2. The mixture was stirred at -78°C for 0.5 hour. Then a solution of cyclobutanone (388.9 mg, 5.6 mmol, 414.6 pL, 1.5 eq) in 20 mL of terahydrofuran was added dropwise at -78°C. The mixture was stirred at -78°C for another 1 hour. After reaction, the mixture was quenched by addition of 100 mL of saturated aqueous NH4CI and extracted with 150 mL of ethyl acetate. The organic phase was separated, washed with 60 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by prep- HPLC (TFA condition) to give 200 mg of compound 2 (764.7 pmol, 20.7% yield) as a yellow oil.

Preparation of compound 3:

To a solution of compound 2 (200 mg, 764.7 pmol, 1 eq ), diphenylmethanimine (166.3 mg, 917.6 pmol, 1.2 eq), t-BuOK (120.1 mg, 1.1 mmol, 1.4 eq) and BINAP (47.6 mg, 76.5 pmol, 0.1 eq) in 4 mL of toluene was added Pd2(dba)3 (7.0 mg, 7.6 pmol, 0.01 eq). The mixture was stirred at 110°C for 5 hours. After reaction, it was filtered, the filtrate was concentrated under reduced pressure. The residue was purified by prep-TLC (S1O2, eluting with a gradient of petroleum ether : ethyl acetate= 3:1) to give 240 mg of compound 3 ( 663.2 pmol, 86.7 % yield) as a light yellow solid.

Preparation of compound 4:

To a solvent of HCI (6 M, 10.0 mL, 108.6 eq ) was added compound 3 (200 mg, 552.7 pmol, 1 eq). The mixture was stirred at 20°C for 4 hours. After reaction, it was adjusted pH to 8~9 with saturated Na2CC>3 and extracted with 5 mL of ethyl acetate, the organic layer was washed with 5mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by prep-TLC (S1O2, eluting with petroleum ether : ethyl acetate=l : l) to give 100 mg of compound 4 (505.9 pmol, 91.5% yield) as a light yellow oil.

Preparation of compound 245:

To a solution of compound 4 (80 mg, 404.7 pmol, 1 eq) and compound 5 (122.7 mg,

445.2 pmol, 1.1 eq) in 2 mL of N,N-Dimethylformamid was added TEA (122.8 mg, 1.2 mmol, 169.0 pL, 3 eq). The mixture was stirred at 20°C for 5 min. Then HATU (184.7 mg, 485.7 pmol, 1.2 eq) was added at 0°C. The mixture was stirred at 30 °C for 12 hours. Then it was partitioned between 10 mL of water and 15mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by prep-TLC (S1O2, eluting with petroleum ether:ethyl acetate= 1 : 1) to give 140 mg of 245 (307.5 pmol, 75.9 % yield) as a white oil. Preparation of compound 253:

245 253

To a solution of 245 (175 mg, 384.3 pmol, 1 eq) in 2 mL of dichloromethane was added BAST (127.5 mg, 576.5 mpioΐ, 126.3 mT, 1.5 eq). The mixture was stirred at 20°C for 12 hours. After reaction, it was washed with 3 mL of saturated aqueous NaHCCL and extracted with 15 mL of dichloromethane. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated in vacuum. The residue was purified by prep- HPLC (TFA condition) to give 15 mg of 253 (30.8 pmol, 8.0% yield, 93.9% purity) as a white solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.75 - 10.69 (m, 1H), 7.91 - 7.87 (m, 1H), 7.80 - 7.73 (m, 2H), 7.68 - 7.65 (m, 1H), 7.59 (s, 2H), 7.56 - 7.52 (m, 1H), 7.20 - 7.16 (m, 1H), 7.12 - 7.08 (m, 1H), 4.00 - 3.89 (m, 2H), 3.47 - 3.38 (m, 2H), 2.83 - 2.75 (m, 1H), 1.76 - 1.57 (m, 4H)

LCMS (ESI+): m/z 457.0 (M+H)

EXAMPLE 40: Synthesis of compound 255:

255 Experimental Procedures:

Preparation of compound 2:

To a solution of l-chloro-3-fluoro-5-nitro-benzene (100.0 mg, 569.7 pmol, 1 eq) in 2 mL of DMSO was added K2CO3 (157.5 mg, 1.1 mmol, 2 eq) and piperidin-4-ol (69.1 mg, 683.6 pmol, 1.2 eq). The mixture was stirred at 80°C for 12 hours. Then it was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 140 mg of crude compound 2 as a yellow oil, which was used directly into the next step.

Preparation of compound 3:

2 3

To a solution of l-(3-chloro-5-nitro-phenyl)piperidin-4-ol (140 mg, 545.4 pmol, 1 eq) in 3 mL of ethyl alcohol and 0.5 mL of water was added Fe (152.3 mg, 2.7 mmol, 5 eq) and NH4CI (145.9 mg, 2.7 mmol, 5 eq). The mixture was stirred at 80°C for 1 hour. Then it was filtered, the filtrate was partitioned between 5 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 120 mg of crude compound 3 as a yellow oil, which was used directly into the next step. Preparation of compound 255:

To a solution of 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoic acid (50 mg, 181.3 pmol, 1.2 eq ) in 2 mL of DMF was added l-(3-amino-5-chloro-phenyl)piperidin-4-ol (49.3 mg, 217.6 pmol, 1.2 eq) and TEA (27.5 mg, 272.0 pmol, 1.5 eq). The mixture was cooled to 0°C,

HATU (103.4 mg, 272.0 pmol, 1.5 eq) was added and stirred at 20°C for 12 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (TFA condition) to give 44.3 mg of 255 (72.5 pmol, 40.0% yield, 98.0% purity, TFA salt) as a white solid.

JH NMR (400MHz, DMSO-d6) d ppm 10.51 (s, 1H), 7.56 (d, J=8.8 Hz, 1H), 7.34 (dd, J=2.9, 8.8 Hz, 1H), 7.29 (d, J=2.9 Hz, 1H), 7.24 - 7.21 (m, 2H), 6.74 (s, 1H), 3.78 (t, J=6.5 Hz, 2H), 3.65 (tt, J=4.1, 8.6 Hz, 1H), 3.58 - 3.48 (m, 4H), 2.96 - 2.87 (m, 2H), 2.42 (quin, J=6.9 Hz, 2H), 1.80 (br dd, J=3.8, 9.3 Hz, 2H), 1.49 - 1.37 (m, 1H), 1.49 - 1.37 (m, 2H)

LCMS (ESI+): m/z 484.1 (M+H)

The following compounds were prepared analogously using the appropriate amine to do SNAr reaction firstly, followed by reduction of nitro group and then do an amide coupling reaction with corresponding acid:

ci

compound 281:

JH NMR (400 MHz, CHLOROF ORM-d) d ppm 8.47 - 8.61 (m, 1 H) 7.62 - 7.66 (m, 1 H) 7.56 -

7.59 (m, 1 H) 7.49 - 7.52 (m, 1 H) 7.31 - 7.33 (m, 1 H) 7.04 - 7.09 (m, 1 H) 3.72 - 3.82 (m, 2 H) 3.42 (br t, / = 7.44 Hz, 2 H) 3.30 - 3.39 (m, 4 H) 2.52 - 2.62 (m, 2 H) 1.87 - 1.98 (m, 4 H) 1.63 - 1.73 (m, 2 H) LCMS (ESI+): m/z 502.0 (M+H)

compound 254:

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 8.39 - 8.48 (m, 1 H) 7.60 - 7.63 (m, 1 H) 7.49 (s, 1 H) 7.45 - 7.47 (m, 1 H) 7.42 (d, / = 1.38 Hz, 2 H) 7.02 - 7.05 (m, 1 H) 3.75 - 3.82 (m, 2 H) 3.38 - 3.44 (m, 2 H) 3.31 - 3.37 (m, 4 H) 2.57 (quin, / = 7.00 Hz, 2 H) 1.87 - 1.94 (m, 4 H) 1.68

(quin, J = 5.78 Hz, 2 H)

LCMS (ESI+): m/z 468.1 (M+H)

F

compound 280:

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 8.47 - 8.58 (m, 1 H) 7.51 - 7.57 (m, 1 H) 7.40 - 7.44 (m, 1 H) 7.24 - 7.27 (m, 1 H) 7.21 (br s, 1 H) 6.98 (s, 1 H) 3.73 - 3.80 (m, 2 H) 3.38 - 3.45

(m, 2 H) 3.28 - 3.35 (m, 4 H) 2.52 - 2.61 (m, 2 H) 1.82 - 1.91 (m, 4 H) 1.61 - 1.72 (m, 2 H)

LCMS (ESI+): m/z 486.0 (M+H)

compound 257:

J H NMR (400 MHz, DMSO -d 6 ) d ppm 10.52 (s, 1 H) 7.56 (d, J = 8.76 Hz, 1 H) 7.32 - 7.36 (m, 1 H) 7.29 (d, J = 2.75 Hz, 1 H) 7.11 (t, / = 1.75 Hz, 1 H) 6.74 (t, / = 1.88 Hz, 1 H) 6.19 (t, / = 1.88 Hz, 1 H) 3.79 - 3.85 (m, 6 H) 3.55 - 3.58 (m, 2 H) 2.43 (t, / = 7.13 Hz, 2 H) 2.28 - 2.34 (m, 2 H)

LCMS (ESI+): m/z 440.0 (M+H)

compound 256: J H NMR (400 MHz, DMSO-<¾) d ppm 10.50 (s, 1 H) 7.56 (d, / = 8.76 Hz, 1 H) 7.33 - 7.37 (m, 1 H) 7.30 (s, 1 H) 7.18 (t, / = 1.69 Hz, 1 H) 7.01 (t, / = 1.94 Hz, 1 H) 6.49 (t, J = 2.06 Hz, 1 H) 3.79 (s, 2 H) 3.57 - 3.59 (m, 2 H) 2.91 (s, 6 H) 2.43 (t, / = 7.13 Hz, 2 H)

LCMS (ESI+): m/z 428.0 (M+H)

compound 273:

J H NMR (400 MHz, METHANOL-^) d ppm 7.49 (br d, / = 8.88 Hz, 1 H) 7.38 (br s, 2 H) 7.08 (br s, 2 H) 6.57 (br s, 1 H) 3.81 (br t, / = 5.63 Hz, 2 H) 3.55 (br s, 4 H) 3.47 (br t, / = 6.94 Hz, 2 H) 3.35 (s, 3 H) 2.99 (s, 3 H) 2.48 - 2.56 (m, 1 H) 2.48 - 2.57 (m, 1 H)

LCMS (ESI+): m/z 472.1 (M+H)

Compound 282:

J H NMR (400MHz, CHLOROFORM-d) d ppm 8.29 (s, 1H), 7.39 (s, 1H), 7.33 (s, 1H), 7.22 (d, / = 2.5 Hz, 1H), 6.85 (s, 1H), 3.97 - 3.88 (m, 1H), 3.71 (t, / = 6.5 Hz, 2H), 3.62 - 3.53 (m, 2H), 3.35 (t, / = 7.4 Hz, 2H), 3.13 - 3.04 (m, 2H), 2.51 (quin, / = 7.0 Hz, 2H), 2.12 - 2.02 (m, 2H), 1.71 (br dd, / = 3.5, 13.1 Hz, 2H)

LCMS (ESI+) : m/z 502.0 (M+H)

Compound 293:

J H NMR (400MHz, CHLOROFORM-d) d ppm 8.02 (s, 1H), 7.30 (dd, / = 2.5, 10.1 Hz, 1H), 7.26 (br s, 1H), 7.20 (s, 1H), 7.12 (s, 1H), 6.54 (s, 1H), 5.22 (quin, J = 5.6 Hz, 1H), 4.99 (t, / = 6.6 Hz, 2H), 4.81 - 4.72 (m, 2H), 3.79 (t, / = 6.5 Hz, 2H), 3.42 (t, / = 7.4 Hz, 2H), 2.65 - 2.54 (m, 2H) LCMS (ESI+): m/z 475.0 (M+H)

Compound 259:

J H NMR (400MHz, DMSO-de) d ppm 10.52 (s, 1 H), 7.56 (d, J = 8.8 Hz, 1H), 7.34 (dd, / = 2.8, 8.8 Hz, 1 H), 7.30 (d, / = 2.8 Hz, 1 H), 7.23 (br d, / = 9.5 Hz, 2 H), 6.75 (s, 1 H), 3.78 - 3.77 (m, 2 H), 3.56 (t, / = 7.3 Hz, 2 H), 3.52 - 3.43 (m, 2 H), 3.36 (dt, / = 4.1, 8.2 Hz, 1 H), 3.26 (s, 3 H),

3.00 - 2.90 (m, 2 H), 2.46 - 2.39 (m, 2 H), 1.91 (br d, / = 10.9 Hz, 2 H), 1.55 - 1.42 (m, 2 H)

LCMS (ESI+): m/z 498.1 (M+H)

Compound 303 was prepared through a different amide coupling condition at the last step: Preparation of compound 303:

303

To a solution of 4-(3-amino-5-chloro-phenoxy)-lH-pyridin-2-one (70 mg, 295.8 pmol, 1 eq ), DIEA (57.3 mg, 443.7 pmol, 1.5 eq ) and compound 2 (97.9 mg, 354.9 pmol, 1.2 eq ) in 1 ml, of dichloromethane was added dropwise T3P (207.1 mg, 325.4 pmol, 50% purity, 1.1 eq) at 0°C. After addition, the resulting mixture was stirred at 25°C for 12 hours. Then it was partitioned between 2 mL of water and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (TFA condition) to 28.6 mg of 303 (57.9 pmol, 19.6% yield, 100% purity) as a yellow solid.

J H NMR (400 MHz, DMSO-de) d ppm 11.48 (br s, 1 H) 10.89 (s, 1 H) 7.72 (t, / = 1.81 Hz, 1 H) 7.58 (d, / = 8.76 Hz, 1 H) 7.49 (t, / = 1.94 Hz, 1 H) 7.43 (d, / = 7.25 Hz, 1 H) 7.36 - 7.39 (m, 1

H) 7.34 - 7.35 (m, 1 H) 7.09 (t, / = 2.06 Hz, 1 H) 6.04 (dd, / = 7.32, 2.56 Hz, 1 H) 5.54 (d, / = 2.50 Hz, 1 H) 3.78 (t, J = 6.44 Hz, 2 H) 3.56 (t, /.= 7.38 Hz, 2 H) 2.42 (quin, J = 6.94 Hz, 2 H)

LCMS (ESI+): m/z 494.1 (M+H) The following compounds were obtained through an ester amide exchange reaction at the last step:

EXAMPLE 41: Compounds synthesized by ester amide exchange reactions:

Preparation of compound 285:

285

To a solution of methyl 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)-3-fluoro-benzoat e (100 mg, 324.9 mpioΐ, 1 eq) and 3-chloro-5-(4-methoxy-l-piperidyl)aniline (93.8 mg, 390.0 pmol, 1.2 eq) in 3 mL of toluene was added AlMe3 (2 M, 324.9 pL, 2 eq) at 0°C. The mixture was heated to 110°C and stirred for 12 hours. After reaction, the mixture was quenched by addition of 10 mL of water, and then extracted with 10 mL of ethyl acetate. The organic layer was separated and washed with 10 mL of brine, separated and concentrated under reduced pressure to get a residue, which was purified by prep- HPLC (TFA condition) to give 39.4 mg of 285 (39.4 mg, 60.8 pmol, 18.7% yield, 97.3% purity, TFA salt) as light yellow solid.

JH NMR (400MHz, DMSO-de) d ppm 10.61 (s, 1 H), 7.33 (dd, J = 2.3, 11.1 Hz, 1 H), 7.21 (br d, / = 10.1 Hz, 3 H), 6.76 (s, 1 H), 3.80 (br t, / = 6.4 Hz, 2 H), 3.63 - 3.61 (m, 2 H), 3.51 - 3.45 (m, 2 H), 3.41 - 3.32 (m, 1 H), 3.26 (s, 3 H), 2.95 (br t, / = 9.8 Hz, 2 H), 2.45 - 2.40 (m, 2 H), 1.91 (br d, / = 10.3 Hz, 2 H), 1.55 - 1.42 (m, 2 H)

LCMS (ESI+): m/z 516.0 (M+H)

Compound 278:

J H NMR (400MHz, DMSO-d6) d ppm 10.62 (s, 1 H), 7.32 (dd, / = 2.5, 11.1 Hz, 1 H), 7.19 (d, J

= 1.5 Hz, 1 H), 7.10 (s, 1H), 6.71 (s, 1 H), 6.23 (t, / = 1.8 Hz, 1 H), 4.56 (quin, / = 5.6 Hz, 1 H), 4.07 (t, 7 = 7.2 Hz, 2 H), 3.80 (t, 7 = 6.5 Hz, 2 H), 3.61 (t, 7 = 7.3 Hz, 2 H), 3.52 (dd, 7 = 4.8, 7.8 Hz, 2 H), 2.46 - 2.39 (m, 2 H)

LCMS (ESI+): m/z 474.0 (M+H)

Compound 283:

JH NMR (400MHz, DMSO-de) d ppm 10.62 (s, 1 H), 7.53 (d, 7 = 2.6 Hz, 1 H), 7.31 (d, 7 = 2.6 Hz, 1 H), 7.22 (br d, 7 = 10.3 Hz, 2 H), 6.78 (s, 1 H), 3.81 (t, 7 = 6.4 Hz, 2 H), 3.61 (t, 7 = 7.3 Hz, 2 H), 3.53 - 3.44 (m, 2 H), 3.41 - 3.33 (m, 1 H), 3.27 (s, 3 H), 3.01 - 2.92 (m, 2 H), 2.43 (quin, 7 = 6.9 Hz, 2 H), 1.97 - 1.86 (m, 2 H), 1.55 - 1.43 (m, 2 H)

LCMS (ESI+): m/z 532.0 (M+H)

Compound 272:

J H NMR (400MHz, DMSO-de) d ppm 10.53 (s, 1 H), 7.56 (d, 7 = 8.8 Hz, 1 H), 7.34 (dd, 7 = 2.8, 8.9 Hz, 1 H), 7.29 (d, 7 = 2.9 Hz, 1 H), 7.12 (s, 1 H), 6.74 (s, 1 H), 6.22 (t, 7 = 1.8 Hz, 1 H), 4.56 (quin, 7 = 5.6 Hz, 1 H), 4.06 (t, 7 = 7.2 Hz, 2 H), 3.78 (t, 7 = 6.5 Hz, 2 H), 3.56 (t, 7 = 7.4 Hz, 2 H), 3.52 (dd, 7 = 4.9, 7.9 Hz, 2 H), 2.42 (t, 7 = 7.0 Hz, 2 H)

LCMS (ESI+): m/z 456.0 (M+H)

Compound 277:

J H NMR (400MHz, DMSO-de) d ppm 10.62 (s, 1 H), 7.52 (d, 7 = 2.6 Hz, 1 H), 7.30 (d, 7 = 2.6 Hz, 1 H), 7.10 (s, 1H), 6.71 (s, 1 H), 6.23 (t, 7 = 1.8 Hz, 1 H), 4.56 (quin, 7 = 5.6 Hz, 1 H), 4.07 (t, 7 = 7.2 Hz, 2 H), 3.81 (t, 7 = 6.5 Hz, 2 H), 3.61 (t, 7 = 7.3 Hz, 2 H), 3.52 (dd, 7 = 4.9, 7.9 Hz, 2 H), 2.42 (t, 7 = 7.0 Hz, 2 H)

LCMS (ESI+): m/z 489.9 (M+H) Compound 307:

J H NMR (400 MHz, DMSO-de) d ppm 10.75 (s, 1 H) 7.63 (s, 1 H) 7.56 (d, J = 8.75 Hz, 1 H) 7.51 (br s, 1 H) 7.42 (dd, J = 9.69, 3.19 Hz, 1 H) 7.35 (dd, J = 8.82, 2.81 Hz, 1 H) 7.29 (d, J = 2.75 Hz, 1 H) 7.17 (s, 1 H) 6.82 (t,/ = 2.06 Hz, 1 H) 6.44 (d, J = 9.76 Hz, 1 H) 3.76 (t, J = 6.44 Hz, 2 H) 3.56 (t, J = 7.32 Hz, 2 H) 2.41 (t, J = 6.94 Hz, 2 H)

LCMS (ESI+): m/z 494.0 (M+H)

EXAMPLE 42: Synthesis of Compound 270:

Experimental Procedures:

Preparation of compound 2:

1 2

To a solution of N-(oxomethylene)sulfamoyl chloride (2 g, 14.1 mmol, 1.2 mL, 1 eq) in 20 mL of dichloromethane was added 2-methylpropan-2-ol (1.3 g, 16.9 mmol, 1.6 mL, 1.2 eq) at 0°C. The mixture was stirred at 0°C for 1 hour. Then it was concentrated under reduced pressure to give 3 g of crude compound 2 as a white solid, which was used into the next step directly. Preparation of compound 3

3

To a solution of 5-amino-2-chloro-benzoic acid (10 g, 58.3 mmol, 1 eq) in 100 mL of methanol was added SOCk (13.9 g, 116.6 mmol, 8.5 mL, 2 eq) at 0°C. The mixture was stirred at 65°C for 4 hours. Then it was concentrated under reduced pressure to give a residue, which was partitioned between 100 mL of saturated NaHCCk and 200 mL of ethyl acetate. The organic phase was separated, washed with 150 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 11 g of crude compound 3 as a yellow oil, which was used into the next step directly.

Preparation of compound 4:

To a solution of methyl 5-amino-2-chloro-benzoate (1 g, 5.4 mmol, 1 eq) in 20 mL of dichloromethane was added TEA (817.8 mg, 8.1 mmol, 1.1 mL, 1.5 eq) and tert-butyl N- chlorosulfonylcarbamate (1.4 g, 6.5 mmol, 1.2 eq) at 0°C. The mixture was stirred at 20°C for 12 hours. Then it was partitioned between 30 mL of saturated NH4CI and 80 mL of ethyl acetate. The organic phase was separated, washed with 20 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =5: 1 to 3: 1) to give 600 mg of compound 4 (1.6 mmol, 30.5% yield) as a white soild. Preparation of compound 5:

To a solution of methyl 5-(tert-butoxycarbonylsulfamoylamino)-2-chloro-benzoate (600 mg, 1.6 mmol, 1 eq) in 10 mL of acetonitrile was added K2CO3 (681.9 mg, 4.9 mmol, 3 eq) and 1 ,2-dibromoethane (926.9 mg, 4.9 mmol, 3 eq). The mixture was stirred at 80°C for 12 hours. Then it was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 600 mg of crude compound 5 as a white solid, which was used into the next step directly.

Preparation of compound 6:

To a solution of tert-butyl 5-(4-chloro-3-methoxycarbonyl-phenyl)- 1,1 -dioxo- 1,2,5- thiadiazolidine-2-carboxylate (600 mg, 1.5 mmol, 1 eq ) in 6 mL of dichloromethane was added TFA (350.1 mg, 3.1 mmol, 2 eq). The mixture was stirred at 20°C for 2 hours. Then it was partitioned between 5 mL of saturated NaHCCb and 15 mL of ethyl acetate. The organic phase was separated, washed with 5mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =4: 1 to 2: 1) to give 330 mg of compound 6 (1.1 mmol, 73.9% yield) as a yellow oil. Preparation of compound 270:

6 270

To a solution of methyl 2-chloro-5-(l,l-dioxo-l,2,5-thiadiazolidin-2-yl)benzoate (40 mg, 137.6 pmol, 1 eq) in 2 mL of toluene was added 3-chloro-5-tetrahydropyran-4-yl-aniline (34.9 mg, 165.1 pmol, 1.2 eq) and AlMe3 (2 M, 137.59 pL, 2 eq) at 0°C. The mixture was stirred at 100°C for 5 hours. The reaction mixture was quenched by 1 mL of methanol, filtered and concentrated under reduced pressure to give a residue, which was purified by prep- HPLC (TFA condition) firstly. The obtained crude product was re-purified by prep- HPLC (neutral condition) again to give 9.9 mg of 270 (18.3 pmol, 13.3% yield, 87.1% purity) as a white solid

JH NMR (400MHz, DMSO-de) d ppm 10.72 (s, 1H), 7.73 (s, 1H), 7.58 (d, / = 8.8 Hz, 1H), 7.53 (s, 1H), 7.34 (dd, / = 2.9, 8.8 Hz, 1H), 7.29 (d, / = 2.8 Hz, 1H), 7.10 (s, 1H), 6.68 (br s, 1H), 3.98 - 3.87 (m, 4H), 3.54 (br t, / = 6.4 Hz, 2H), 3.45 (br s, 2H), 2.90 - 2.74 (m, 1H), 1.76 - 1.56 (m, 4H)

LCMS (ESI+): m/z 470.0 (M+H)

The following compounds were prepared analogously coupling the appropriate ester (or acid) with corresponding aniline:

J H NMR (400 MHz, DMSO-rfc) d ppm 10.56 (s, 1 H) 7.91 (br s, 1 H) 7.57 (d, / = 8.88 Hz, 1 H) 7.33 (dd, J = 8.88, 2.50 Hz, 1 H) 7.29 (s, 1 H) 7.26 (d, J = 2.38 Hz, 1 H) 7.23 (s, 1 H) 6.77 (s, 1

H) 3.89 (br t, / = 6.25 Hz, 2 H) 3.71 - 3.74 (m, 4 H) 3.53 (br d, / = 5.38 Hz, 2 H) 3.12 (br d, / = 4.25 Hz, 4 H)

LCMS (ESI+): m/z 471.0 (M+H)

6 6A

To a solution of methyl 2-chloro-5-(l,l-dioxo-l,2,5-thiadiazolidin-2-yl)benzoate (300 mg, 1.0 mmol, 1 eq ) in 3 mL of acetonitrile was added CS2CO3 (437.1 mg, 1.3 mmol, 1.3 eq ) and Mel (175.8 mg, 1.2 mmol, 1.2 eq). The mixture was stirred at 20°C for 2 hours. Then it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 300 mg of crude compound 6A as a yellow solid, which was used into the next step directly.

The obtained ester was then coupled with appropriate aniline to give 252.

J H NMR (400MHz, CHLOROFORM-d) d ppm 8.16 (s, 1H), 7.61 (s, 1H), 7.45 (s, 3H), 7.39 (s, 1H), 7.03 (s, 1H), 4.08 (br d, / = 11.1 Hz, 2H), 3.83 (t, / = 6.2 Hz, 2H), 3.56 - 3.46 (m, 4H), 2.86 (s, 3H), 2.81 - 2.71 (m, 1H), 1.84 - 1.73 (m, 4H)

LCMS (ESI+) : m/z 484.1 (M+H)

Experimental procedure of amide coupling reaction:

297 To a solution of 2-chloro-3-fluoro-5-(5-methyl-l, l-dioxo-l,2,5-thiadiazolidin-2- yl)benzoic acid (100 mg, 323.9 mihoI, 1 eq ) in 3 mL of dimethyl form amide was added 3-chloro- 5-tetrahydropyran-4-yl-aniline (75.4 mg, 356.3 mihoI, 1.1 eq) and TEA (39.3 mg, 388.7 mitioI,

I.2 eq). The mixture was cooled to 0°C, HATU (147.8 mg, 388.7 mihoI, 1.2 eq) was added and stirred at 20°C for 4 hours. Then it was filtered, the fitrate was purified by prep- HPLC (TFA condition) to give 297 (14.6 mg, 28.3 mihoI, 8.7 % yield, 97.3% purity) as a white solid.

J H NMR (400MHz, DMSO-cfc) d ppm 10.81 (s, 1H), 7.71 (s, 1H), 7.50 (s, 1H), 7.35 (dd, / = 2.6,

I I.0 Hz, 1H), 7.23 (s, 1H), 7.12 (s, 1H), 3.98 - 3.89 (m, 4H), 3.54 (t, / = 6.4 Hz, 2H), 3.42 (br t, J = 10.7 Hz, 2H), 2.83 - 2.77 (m, 1H), 2.76 (s, 3H), 1.76 - 1.55 (m, 4H)

LCMS (ESI+) : m/z 502.0 (M+H)

EXAMPLE 43: Synthesis of compound 271:

Experimental Procedures:

Preparation of compound 3:

3A 3 To a solution of 3-chloro-5-iodo-benzoic acid (4.5 g, 15.9 mmol, 1 eq) in 45 mL of t- BuOH was added dropwise DPPA (6.6 g, 23.9 mmol, 5.2 mL, 1.5 eq), TEA (2.4 g, 23.9 mmol, 3.3 mL, 1.5 eq) at 20°C. After addition, the mixture was stirred at this temperature for 1 hr, and then warmed to 80°C and stirred for another 12 hrs. After reaction, it was concentrated under reduced pressure to give the crude product, which was purified by column chromatography (Si02, eluting with a gradient of petroleum ether : ethyl acetate =1 :0 to 1 :0) to give 2.7 g of compound 3 (7.7 mmol, 48.5% yield) as a yellow oil.

Preparation of compound 2:

To a solution of 1-methylcyclobutanecarboxylic acid (800 mg, 7.0 mmol, 1.0 eq) in 10 mL of dichloromethane was added 2-hydroxyisoindoline-l,3-dione (1.1 g, 7.0 mmol, 1.0 eq), EDCI (1.5 g, 7.7 mmol, 1.1 eq) and DMAP (85.6 mg, 700.9 pmol, 0.1 eq). The mixture was stirred at 25 °C for 4 hrs. Then it was partitioned between 51 mL of water and 35 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (Si02, eluting with a gradient of petroleum ether : ethyl acetate =10: 1 to 5:1) to give 1.8 g of compound 2 (6.8 mmol, 97.4% yield) as a yellow oil.

Preparation of compound 4:

A mixture of tert-butyl N-(3-chloro-5-iodo-phenyl)carbamate (1.5 g, 4.2 mmol, 1.0 eq), (l,3-dioxoisoindolin-2-yl) 1-methylcyclobutanecarboxylate (1.2 g, 4.7 mmol, 1.1 eq), Zn (554.8 mg, 8.5 mmol, 2 eq), Ni(dtbbpy)Br2 (413.1 mg, 848.5 pmol, 0.2 eq) in 6 mL of DMA was degassed and purged with N2 for 3 times, and then the mixture was stirred at 40°C for 12 hours under N2 atmosphere. Then it was partitioned between 15 mL of water and 35 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =20:1 to 10: 1) to give 200 mg of compound 4 (676.1 pmol, 15.9% yield) as a yellow oil.

Preparation of compound 5:

To a solution of tert-butyl N-[3-chloro-5-(l-methylcyclobutyl)phenyl]carbamate (200 mg, 676.1 pmol, 1 eq ) in 2 mL of dichloromethane was added TFA (924.0 mg, 8.1 mmol, 0.6 mL, 12.0 eq). The mixture was stirred at 20°C for 1 hour. Then it was partitioned between 3 mL of saturated NaHCCb and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 130 mg of the crude compound 3 as a yellow oil.

Preparation of compound 271:

To a solution of 3-chloro-5-(l-methylcyclobutyl)aniline (30 mg, 153.3 pmol, 1 eq) in 2 mL of toluene was added methyl 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoate (48.8 mg, 168.6 pmol, 1.1 eq) and A1(C]¾) 3 (2 M, 114.98 pL, 1.5 eq) at 0°C. The mixture was stirred at

100°C for 4 hours. Then it was quenched by addition of 1 mL of methanol, filtered and the filtrate was concentrated under reduced pressure to give a residue, which was purified by prep- HPLC (TFA condition) twice to give 8.2 mg of 271 (17.6 pmol, 11.5% yield, 97.3% purity) as a white solid. J H NMR (400MHz, CHLOROFORM-c/) d ppm 7.99 (br s, 1H), 7.61 (s, 1H), 7.53 - 7.48 (m, 1H), 7.48 - 7.42 (m, 2H), 7.21 (s, 1H), 6.96 (s, 1H), 3.80 (t, / = 6.5 Hz, 2H), 3.42 (t, J = 7.5 Hz, 2H), 2.58 (quin, / = 6.9 Hz, 2H), 2.42 - 2.32 (m, 2H), 2.17 - 2.03 (m, 3H), 1.91 - 1.79 (m, 1H), 1.46 (s, 3H)

LCMS (ESI+): m/z 453.0 (M+H)

Compound 275 was prepared analogously coupling the appropriate acid with the aniline 5:

compound 275

J H NMR (400MHz, CHLOROFORM-d) d ppm 7.81 (br s, 1H), 7.60 (s, 1H), 7.54 (d, J = 2.6 Hz, 1H), 7.35 (d, J = 2.5 Hz, 1H), 7.19 (s, 1H), 6.96 (s, 1H), 3.77 (t, J = 6.5 Hz, 2H), 3.42 (t, J =7.4 Hz, 2H), 2.58 (quin, / = 6.9 Hz, 2H), 2.41 - 2.30 (m, 2H), 2.12 - 2.00 (m, 3H), 1.90 - 1.79 (m, 1H), 1.46 (s, 3H)

LCMS (ESI+): m/z 489.0 (M+H)

EXAMPLE 44: Synthesis of compound 294:

Experimental Procedures:

Preparation of compound 2:

To a solution of l,3-dichloro-5-nitro-benzene (1 g, 5.2 mmol, 2.0 mL, 1 eq) in 2 mL of dimethyl sulfoxide was added K2CO3 (1.4 g, 10.4 mmol, 2 eq) and piperidin-4-ol (1.1 g, 10.4 mmol, 2 eq). The mixture was stirred at 110°C for 12 hours. Then it was partitioned between 5 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with petroleum ether : ethyl acetate = 1 : 1) to give 590 mg of compound 2 (2.3 mmol, 44.1% yield) as a yellow solid. Preparation of compound 3

To a solution of compound 2 (400 mg, 1.6 mmol, 1 eq ) in 4 mL of dimethyl form amide was added NaH (124.7 mg, 3.1 mmol, 60% purity, 2 eq) at 0°C. After addition, the mixture was stirred at this temperature for 1 hour, and then 2-bromoethoxy-tert-butyl-dimethyl-silane (745.6 mg, 3.1 mmol, 2 eq) was added dropwise at 20°C. The mixture was stirred at 20°C for 11 hours. Then it was partitioned between 10 mL of water and 20 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate = 100: 1 to 1: 1) to give 190 mg of compound 3 (457.8 pmol, 29.4% yield) as a yellow oil.

Preparation of compound 4:

The mixture of compound 3 (190 mg, 457.8 pmol, 1 eq), Fe (127.8 mg, 2.3 mmol, 5 eq) and NH4CI (122.5 mg, 2.3 mmol, 5 eq) in 2 mL of ethanol and 2 mL of water was degassed and purged with N2 or 3 times, and then the mixture was stirred at 80°C for 12 hours under N2 atmosphere. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was partitioned between 3 mL of water and 5 mL of ethyl acetate. The organic phase was separated, washed with 6 mL of water and 2 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 160 mg of crude compound 4 as a yellow oil, which was used directly into the next step without purification. Preparation of compound 6:

To a solution of 4 (160 mg, 415.6 pmol, 1 eq ), TEA (126.2 mg, 1.3 mmol, 173.5 pL, 3 eq ) and 2,3-dichloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoic acid (193.3 mg, 623.4 pmol, 1.5 eq) in 2 mL of dimethyl form amide was added dropwise HATU (316.0 mg, 831.1 pmol, 2 eq) in 0.5 mL of dimethyl form amide at 0°C. After addition, the mixture was stirred at 20°C for 12 hours. Then it was partitioned between 5 mL of water and 8 mL of ethyl acetate. The organic phase was separated, washed with 6 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with petroleum ether :ethyl acetate = 1 : 1) to give 200 mg of compound 6 (295.4 pmol, 71.1% yield) as a yellow solid.

A mixture of compound 6 (0.2 g, 295.4 pmol, 1 eq) and TLA (770.0 mg, 6.8 mmol, 500.0 pL, 22.9 eq) in 2 mL of dichloromethane was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20°C for 12 hours under N2 atmosphere. After reaction, it was concentrated in vacuum to give 250 mg of crude compound 7 (TLA salt) as a yellow oil, which was used directly into the next step without purification. Preparation of compound 294:

A mixture of compound 7 (250 mg, 323.5 pmol, 1 eq, TFA salt) and NaOH (25.9 mg, 646.9 pmol, 2 eq) in 2 mL of MeOH and 0.5 mL of H2O was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20°C for 1 hour under N2 atmosphere. After reaction, it was filtered and the filter cake was washed three times with 3 mL of water, then the filter cake was dried in vacuum to give 62.7 mg of 294 (109.7 pmol, 33.9% yield, 98.5% purity) as a yellow solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.60 (s, 1 H), 7.52 (d, / = 2.50 Hz, 1 H), 7.31 (d, / = 2.50 Hz, 1 H), 7.19 (br d, / = 8.88 Hz, 2 H), 6.75 (s, 1 H), 4.56 (t, / = 5.19 Hz, 1 H), 3.81 (t, / = 6.44 Hz, 2 H), 3.60 (t, / = 7.32 Hz, 2 H), 3.43 - 3.54 (m, 7 H), 2.94 (br t, / = 9.94 Hz, 2 H), 2.39 - 2.45 (m, 2 H), 1.90 (br d, J= 10.13 Hz, 2 H), 1.45 - 1.55 (m, 2 H).

LCMS (ESI+): m/z 564.1 (M+H)

EXAMPLE 45: Synthesis of compound 295:

Experimental Procedures:

The synthesis of compound 2 can be found in the synthesis of compounds 224, 225.

Preparation of compound 1:

To a solution of 3-bromo-5-chloro-aniline (10 g, 48.4 mmol, 1 eq), 4, 4, 5, 5- tetramethyl- 2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxab orolane (13.5 g, 53.3 mmol, 1.1 eq) and KOAc (9.5 g, 96.9 mmol, 2 eq) in 100 mL of dioxane was added Pd(dppf)Cl2 (3.5 g, 4.8 mmol, 0.1 eq) under N2. The mixture was stirred at 110°C for 12 hrs. Then it was partitioned between 100 mL of water and 150 mL of ethyl acetate. The organic phase was separated, washed with 100 mL of brine, dried over Na 2 S0 4 , filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether/ethyl acetate=20: 1 to 3 : 1) to give 9.2 g of compound 1 (36.1 mmol, 74.6% yield) as a yellow oil.

Preparation of compound 3:

To a solution of compound 1 (3 g, 11.8 mmol, 1 eq ), compound 2 (7.7 g, 35.5 mmol, 3 eq ) and CS2CO3 (7.7 g, 23.7 mmol, 2 eq) in 40 mL of dioxane and 8 mL of H2O was added Pd(dppf)Cl2 (865.8 mg, 1.2 mmol, 0.1 eq) under N2. The mixture was stirred at 100°C forl2 hours. Then it was partitioned between 40 mL of water and 60 mL of ethyl acetate. The organic phase was separated, washed with 30 mL of brine, dried over Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate=20:l to 1:1) to give 900 mg of compound 3 (4.6 mmol, 38.9% yield) as a yellow oil.

Preparation of compound 4:

To a solution of compound 3 (400 mg, 2.0 mmol, 1 eq) in 50 mL of ethyl acetate was added Pd/C (200 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with H2 several times. The mixture was stirred at 20°C for 2 hours under H2 (15psi) atmosphere. After reaction, it was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep- HPLC (TFA condition) to give 220 mg of racemic compound 4 (705.8 pmol, 34.5% yield, TFA salt) as a brown oil. Preparation of compound 4B, 4C:

The racemic compound 4 (220 mg, 705.8 pmol, TFA salt) was separated by SFC (column: DAICEL CF1IRALPAK AY-F1 (250mm*30mm, 5pm); mobile phase: [0.1 %N¾H2q EtOFl]; B %: 23%-23%, 5min) to give 80 mg of compound 4B (404.7 pmol, 57.3% yield) and 80 mg of compound 4C (404.7 pmol, 57.3% yield) as a yellow oil.

The structures of the two isomers were random assignment.

Preparation of compound 295:

To a solution of compound 5 (138.1 mg, 445.2 pmol, 1.1 eq ) in 2 mL of DMF was added compound 4B (80 mg, 404.7 pmol, 1 eq) and TEA (122.9 mg, 1.2 mmol, 169.0 pL, 3 eq). The mixture was cooled to 0°C, F1ATU (230.8 mg, 607.1 pmol, 1.5 eq) was added and stirred at 20°C for 12 hours. Then it was purified by prep- F1PLC (TFA condition) to give 52.9 mg of 295 (108.0 pmol, 26.7% yield, 100% purity) as a light yellow solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.87 - 10.75 (m, 1H), 7.80 - 7.70 (m, 1H), 7.59 - 7.46 (m, 2H), 7.39 - 7.29 (m, 1H), 7.18 - 7.09 (m, 1H), 4.06 - 3.88 (m, 2H), 3.86 - 3.74 (m, 3H), 3.66 - 3.53 (m, 3H), 3.45 - 3.37 (m, 1H), 2.48 - 2.40 (m, 2H), 2.38 - 2.28 (m, 1H), 1.95 - 1.82 (m,

1H).

LCMS (ESI+): m/z 489.0 (M+H)

296 was prepared analogously coupling the acid 5 with the aniline 4C. compound 296:

J H NMR (400 MHz, DMSO-de) d ppm 10.85 - 10.77 (m, 1H), 7.78 - 7.72 (m, 1H), 7.58 - 7.47 (m, 2H), 7.37 - 7.29 (m, 1H), 7.18 - 7.10 (m, 1H), 4.05 - 3.89 (m, 2H), 3.87 - 3.73 (m, 3H), 3.66

- 3.53 (m, 3H), 3.46 - 3.37 (m, 1H), 2.48 - 2.39 (m, 2H), 2.38 - 2.28 (m, 1H), 1.95 - 1.81 (m,

1H).

LCMS (ESI+): m/z 489.0 (M+H)

EXAMPLE 46: Synthesis of compound 260:

260

Experimental procedures:

Preparation of compound 2:

To a solution of oxetan-3-ol (46.4 mg, 626.6 pmol, 1.1 eq) in 2 mL of DMF was added NaH (68.4 mg, 1.7 mmol, 60% purity, 3 eq) at 0°C. After addition, the mixture was stirred at this temperature for 0.5 hour, and then compound 1 (100 mg, 569.6 pmol, 1 eq) was added dropwise at 0°C. The resulting mixture was stirred at 20°C for 0.5 hour. After reaction, it was diluted with 20 mL of sat.NfpCI and extracted with 50 mL of ethyl acetate. The combined organic layers were washed with 50 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 130 mg of crude compound 2 as a yellow oil, which was used directly into the next step without purification.

Preparation of compound 3:

To a solution of compound 2 (120 mg, 522.6 pmol, 1 eq) in 2 mL of ethyl alcohol and 0.5 mL of water was added Fe (145.9 mg, 2.6 mmol, 5 eq) and NH4CI (139.8 mg, 2.6 mmol, 5 eq) . The mixture was stirred at 80°C for 12 hours. After reaction, it was filtered and concentrated under reduced pressure to remove ethanol. The residue was diluted with 50 mL of water and extracted with 100 mL of ethyl acetate. The organic layer was washed with 50 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to get 104 mg of crude compound 3 as a yellow oil, which was used directly into the next step without purification.

Preparation of compound 260:

To a solution of 3-chloro-5-(oxetan-3-yloxy)aniline (50 mg, 250.5 pmol, 1 eq) and methyl 2,3-dichloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoate (81.2 mg, 250.5 pmol, 1 eq) in 2 mL of toluene was added dropwise AlMe3 (2 M, 250.5 pL, 2 eq) at 0°C. The mixture was heated to 110°C and stirred for 12 hours. After reaction, it was concentrated under reduced pressure to remove toluene. The reaction mixture was quenched by addition of 2 mL of methanol. Then the mixture was filtered and the filtrate was purified by prep- HPLC (TFA condition) to give 4.8 mg of 260 (8.7 pmol, 3.5% yield, 89.2% purity) as a brown solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.81 - 10.83 (m, 1 H), 7.53 - 7.56 (m, 1 H), 7.46 - 7.49 (m, 1 H), 7.33 (d, / = 2.63 Hz, 1 H), 7.09 - 7.11 (m, 1 H), 6.67 (s, 1 H), 5.31 (quin, / = 5.44 Hz,

1 H), 4.89 - 4.94 (m, 2 H), 4.55 (dd, J = 7.13, 5.00 Hz, 2 H), 3.82 (t, / = 6.44 Hz, 2 H), 3.61 (t, J = 7.25 Hz, 2 H), 2.41 - 2.46 (m, 2 H).

LCMS (ESI+): m/z 493.0 (M+H)

Compound 258 was prepared analogously using tetrahydrofuran-3-ol to do a SNAr reaction at the first step, followed by reduction of the nitro group and then do an ester amide exchange reaction.

compound 258:

J H NMR (400 MHz, DMSO-de) d ppm 10.78 (s, 1 H), 7.54 (d, / = 2.63 Hz, 1 H), 7.43 (s, 1 H), 7.33 (d, / = 2.63 Hz, 1 H), 7.22 (s, 1 H), 6.82 (t, J = 1.75 Hz, 1 H), 5.01 - 5.08 (m, 1 H), 3.72 - 3.92 (m, 6 H), 3.61 (t, / = 7.38 Hz, 2 H), 2.44 (t, / = 7.00 Hz, 2 H), 2.18 - 2.28 (m, 1 H), 1.93 - 2.01 (m, 1 H).

LCMS (ESI+): m/z 507.0 (M+H)

EXAMPLE 47: Synthesis of compound 269:

Experimental Procedures:

Preparation of compound 2:

To a solution of l,3-dichloro-5-nitro-benzene (0.2 g, 1.0 mmol, 400.0 pL, 1 eq) in 2 mL of dimethylsulfoxide was added K2CO3 (287.9 mg, 2.1 mmol, 2 eq) and N-methylmethanamine (293.5 mg, 2.6 mmol, 329.8 pL, 2.5 eq) (40% purity). The mixture was stirred at 110°C for 12 hours. Then it was partitioned between 2 mL of water and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with petroleum ether : ethyl acetate = 10: 1) to give 140 mg of compound 2 (697.8 pmol, 67.0% yield) as a yellow solid.

Preparation of compound 3:

A mixture of 3-chloro-N,N-dimethyl-5-nitro-aniline (140 mg, 697.8 pmol, 1 eq), Fe (194.8 mg, 3.5 mmol, 5 eq) and NH4CI (224.0 mg, 4.2 mmol, 6 eq) in 2 mL of EtOH and 0.5 mL of water was degassed and purged with N2 for 3 times, and then the mixture was stirred at 80°C for 12 hours under N2 atmosphere. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. Then it was partitioned between 2 mL of water and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 110 mg of crude compound 3 as a yellow oil, which was used directly into the next step without purification. Preparation of compound 269:

To a solution of 2,3-dichloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoic acid (50 mg, 161.2 pmol, 1 eq ), TEA (48.9 mg, 483.6 pmol, 67.3 pL, 3 eq ) and 5-chloro-Nl,Nl-dimethyl- benzene- 1,3-diamine (35.8 mg, 209.6 pmol, 1.3 eq) in 1 mL of dimethyl formamide was added dropwise HATU (92.0 mg, 241.8 pmol, 1.5 eq) in 1 mL of dimethyl formamide at 0 °C. After addition, the resulting mixture was stirred at 20°C for 12 hours. The reaction mixture was filtered, the filtrate was purified by prep- HPLC (TFA condition) to give 38.5 mg of compound 269 (66.8 pmol, 41.4% yield, 100% purity, TFA salt) as a yellow solid.

JH NMR (400 MHz, DMSO-de) d ppm 10.60 (s, 1 H) 7.52 (d, J = 2.63 Hz, 1 H) 7.30 (d, / = 2.63 Hz, 1 H) 7.14 (s, 1 H) 6.96 (s, 1 H) 6.50 (s, 1 H) 3.81 (t, / = 6.44 Hz, 2 H) 3.60 (t, / = 7.32 Hz, 2 H) 2.90 (s, 6 H) 2.42 (br t, / = 6.94 Hz, 2 H)

LCMS (ESI+): m/z 462 (M+H)

Compound 286 was prepared analogously using the appropriate amine to do SNAr reaction firstly, followed by reduction of the nitro group and then do an amide coupling reaction with corresponding acid:

J H NMR (400 MHz, METH AN OL-CIA) d ppm 7.30 (dd, / = 10.94, 2.56 Hz, 1 H) 7.21 (s, 1 H) 7.11 (br d, / = 10.76 Hz, 2 H) 6.63 (s, 1 H) 3.82 (t, / = 6.50 Hz, 2 H) 3.55 (s, 4 H) 3.50 (t, J = 7.38 Hz, 2 H) 3.35 (s, 3 H) 3.01 (s, 3 H) 2.53 (quin, J = 6.94 Hz, 2 H)

LCMS (ESI+): m/z 490.0 (M+H) EXAMPLE 48: Synthesis of compound 299:

To a solution of l-chloro-3-methoxy-5-nitro-benzene (10 g, 53.3 mmol, 1 eq) in 100 mL of dichloromethane was added dropwise BBr3 (26.7 g, 106.6 mmol, 10.3 mL, 2 eq) at 0°C. After addition, the resulting mixture was stirred at 20°C for 12 hours. Then it was partitioned between 300 mL of saturated NaHCCL and 500 mL of dichloromethane. The organic phase was separated, washed with 300 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (SiCh, eluting with petroleum ether : ethyl acetate = 100: 1 to 10: 1) to give 6.9 g of compound 2 (39.8 mmol, 74.6% yield) as a yellow solid. Preparation of compound 3:

To a solution of 3-chloro-5-nitro-phenol (200 mg, 1.2 mmol, 1 eq) in 3 mL of dimethylsulfoxide was added K2CO3 (318.5 mg, 2.3 mmol, 2 eq) and 2-fluoropyrazine (135.6 mg, 1.4 mmol, 1.2 eq). The mixture was stirred at 70°C for 12 hours. Then it was partitioned between 10 mL of water and 35 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 280 mg of crude compound 3 as a yellow oil, which was used into the next step directly.

Preparation of compound 4:

3 4

To a solution of 2-(3-chloro-5-nitro-phenoxy) pyrazine (280 mg, 1.1 mmol, 1 eq) in 5 mL of ethyl alcohol and 1 mL of water was added NH4CI (297.6 mg, 5.6 mmol, 5 eq) and Fe (310.7 mg, 5.6 mmol, 5 eq). The mixture was stirred at 80°C for 1 hour. Then it was filtered and the filtrate was concentrated under reduced pressure to give a residue, which was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 240 mg of crude compound 4 as a yellow oil, which was used into the next step directly.

Preparation of compound 299:

4 299

To a solution of 3-chloro-5-pyrazin-2-yloxy-aniline (100 mg, 451.1 pmol, 1 eq) in 3 mL of dimethyl formamide was added compound 5 (124.4 mg, 451.2 pmol, 1 eq) and TEA (54.8 mg, 541.4 pmol, 1.2 eq). The mixture was cooled to 0°C, HATU (205.9 mg, 541.4 pmol, 1.2 eq) was added and stirred at 20°C for 12 hours. Then it was filtered, the filtrate was purified by prep-HPLC (TFA condition) to give 69.4 mg of 299 (144.5 pmol, 32.0% yield, 99.8% purity) as a white solid.

J H NMR (400MHz, DMSO-de) d ppm 10.87 (s, 1H), 8.61 (s, 1H), 8.44 (d, 7 =2.7 Hz, 1H), 8.27 (s, 1H), 7.72 (s, 1H), 7.58 (d, 7 = 8.7 Hz, 1H), 7.53 (s, 1H), 7.39 - 7.32 (m, 2H), 7.18 - 7.13 (m, 1H), 3.78 (t, 7 = 6.5 Hz, 2H), 3.56 (t, 7 = 7.3 Hz, 2H), 2.42 (quin, 7 = 6.9 Hz, 3H)

LCMS (ESI+): m/z 479.0 (M+H)

The following compounds were prepared analogously using different halides to do SNAr reaction at the second step.

compound 301

J H NMR (400 MHz, DMSO-de) d ppm 10.83 - 10.94 (m, 1 H) 8.81 (s, 1 H) 8.73 (d, 7 = 5.75 Hz, 1 H) 7.75 (s, 1 H) 7.52 - 7.61 (m, 2 H) 7.32 - 7.39 (m, 2 H) 7.24 (d, 7 = 5.75 Hz, 1 H) 7.20 (s, 1 H) 3.78 (br t, 7 = 6.44 Hz, 2 H) 3.56 (t, 7 = 7.38 Hz, 2 H) 2.38 - 2.45 (m, 2 H)

LCMS (ESI+): m/z 479.0 (M+H)

compound 298 J H NMR (400 MHz, DMSO-de) d ppm 10.92 - 10.81 (m, 1H), 8.76 - 8.64 (m, 2H), 7.78 - 7.69 (m, 1H), 7.61 - 7.49 (m, 2H), 7.42 - 7.27 (m, 3H), 7.20 - 7.12 (m, 1H), 3.85 - 3.71 (m, 2H), 3.64 - 3.49 (m, 2H), 2.46 - 2.38 (m, 2H)

LCMS (ESI+): m/z 479.0 (M+H)

EXAMPLE 49: Synthesis of Compound 300:

Experimental Procedures:

Preparation of compound 2:

1 2

A mixture of pyrimidin-5-ylboronic acid (0.2 g, 1.6 mmol, 1 eq ) and H2O2 (219.6 mg, 1.9 mmol, 186.1 pL, 30% purity, 1.2 eq) in 2 mL of methanol was degassed and purged with N2 for 3 times, and then the mixture was stirred at 25 °C for 1 hour under N2 atmosphere. The reaction mixture was concentrated in vacuum to give 160 mg of crude compound 2 as a yellow solid, which was used directly into the next step without purification. Preparation of compound 3:

2 3

A mixture of pyrimidin-5-ol (160 mg, 1.7 mmol, 1 eq ), 1-chloro- 3-fluoro-5-nitro- benzene (292.3 mg, 1. 7 mmol, 1 eq) and CS2CO3 (1.4 g, 4.2 mmol, 2.5 eq) in 2 mL of dimethylsulfoxide was degassed and purged with N2 for 3 times, and then the mixture was stirred at 110°C for 12 hours under N2 atmosphere. Then it was partitioned between 5 mL of water and 10 mL of ethyl acetate. The organic phase was separated, washed with 8 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 0.3 g of crude compound 3 as a yellow oil, which was used directly into the next step without purification.

Preparation of compound 4:

3 4

A mixture of 5-(3-chloro-5-nitro-phenoxy)pyrimidine (0.3 g, 1.2 mmol, 1 eq), Fe (332.9 mg, 6.0 mmol, 5 eq) and NH4CI (318.9 mg, 6.0 mmol, 5 eq) in 3 mL of ethyl alcohol and 1 mL of H2O was degassed and purged with N2 for 3 times, and then the mixture was stirred at 80°C for 12 hours under N2 atmosphere. After reaction, it was concentrated under reduced pressure to remove ethyl alcohol, then it was partitioned between 5 mL of water and 10 mL of ethyl acetate. The organic phase was separated, washed with 8 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 240 mg of crude compound 4 as a yellow oil, which was used directly into the next step without purification. Preparation of compound 300:

To a solution of 3-chloro-5-pyrimidin-5-yloxy-aniline (120 mg, 541.4 pmol, 1 eq), TEA (164.4 mg, 1.6 mmol, 226.1 pL, 3 eq) and 2-chloro-5-(l,l-dioxo- 1 ,2-thiazolidin-2-yl)benzoic acid (149.3 mg, 541.4 pmol, 1 eq) in 2 mL of DMF was added dropwise HATU (308.79 mg, 812.12 umol, 1.5 eq) in 0.5 mL of DMF at 0°C. After addition, the resulting mixture was stirred at 25 °C for 12 hrs. Then it was filtered. The filtrate was purified by prep- HPLC (TFA condition) to give 23.6 mg of 300 (47.6 pmol, 8.8% yield, 96.7% purity) as a yellow solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.83 (br s, 1 H) 9.07 (s, 1 H) 8.76 (s, 2 H) 7.70 (br s, 1 H) 7.57 (br d, / = 8.63 Hz, 1 H) 7.36 (br s, 2 H) 7.31 (br s, 1 H) 7.05 (br s, 1 H) 3.77 (br t, / = 5.75 Hz, 2 H) 3.55 (br t, / = 7.13 Hz, 2 H) 2.39 - 2.44 (m, 2 H)

LCMS (ESI+): m/z 479.0 (M+H)

The following compounds were prepared analogously to compound 300 coupling the acid 5 with the appropriate aniline.

compound 234:

J H NMR (400 MHz, CHLOROF ORM-c/) d ppm 8.17 (br s, 1 H) 7.53 (s, 1 H) 7.42 - 7.49 (m, 4 H) 6.96 (s, 1 H) 6.35 - 6.75 (m, 1 H) 3.78 (t, J = 6.54 Hz, 2 H) 3.40 (t, J = 7.46 Hz, 2 H) 2.57 (quin, J = 6.97 Hz, 2 H)

LCMS (ESI+): m/z 468.0 (M+H)

Synthesis of the aniline of 234

Experimental Procedures:

Preparation of compound 2:

1 2

To a solution of l-chloro-3-methoxy-5-nitro-benzene (5 g, 26.7 mmol, 1 eq) in 30 mL of dichloromethane was added BBb (20.0 g, 80.0 mmol, 7.7 mL, 3 eq) at 0°C. The mixture was stirred at 15°C for 12 hours. Then it was partitioned between 30 mL of water and 50 mL of ethyl acetate. The organic phase was separated, washed with 15mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =100:1 to 30: 1) to give 4.3 g compound 2 (24.8 mmol, 93.0% yield) as a white solid.

Preparation of compound 3:

A mixture of 3-chloro-5-nitro-phenol (1 g, 5.8 mmol, 1 eq), Fe (1.6 g, 28.8 mmol, 5 eq) and NH4CI (3.1 g, 57.6 mmol, 10 eq) in 10 mL of methanol and 4 mL of water was degassed and purged with N2 for 3 times, and then the mixture was stirred at 70°C for 12 hours under N2 atmosphere. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. Then it was partitioned between 2 mL of water and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 1 g of crude compound 3 as a yellow oil, which was used directly into the next step without purification. Preparation of compound 4:

To a solution of 3-amino-5-chloro-phenol (0.1 g, 696.5 mihoI, 1 eq) and KOH (390.8 mg, 7.0 mmol, 10 eq) in 1 mL of acetonitrile and 1 mL of water was added drop wise 1- [[bromo(difluoro)methyl]-ethoxy-phosphoryl]oxye thane (223.2 mg, 835.8 pmol, 1.2 eq) at -

78°C. After addition, the resulting mixture was stirred at 20°C for 1 hour. Then it was partitioned between 2 mL of water and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (SiCh, eluting with petroleum ether : ethyl acetate = 2: 1) to give 25 mg of compound 4 (129.2 pmol, 18.5% yield) as a yellow oil.

Then compound 4 was used into an amide coupling reaction by using a normal method to give

234.

compound 239: J H NMR (400MHz, CHLOROFORM-d) d ppm 8.44 (br s, 1H), 7.81 (br d, / = 11.7 Hz, 2H), 7.53 - 7.41 (m, 3H), 7.22 (s, 1H), 6.81 - 6.34 (m, 1H), 3.80 (t, J = 6.5 Hz, 2H), 3.41 (t, J = 7.5 Hz, 2H), 2.58 (quin, / = 7.0 Hz, 2H)

LCMS (ESI+): m/z 442.0 (M+H)

Synthesis of the aniline of 239

4

Experimental Procedures:

Preparation of compound 2:

A mixture of 3-bromo-5-methoxy-aniline (200 mg, 989.8 pmol, 1 eq), Pd(OAc)2 (22.2 mg, 98.9 pmol, 0.1 eq), K4[Fe(CN)6] (218.7 mg, 593.92 pmol, 0.6 eq) and Na2C(¼ (209.8 mg, 1.98 mmol, 2 eq) in 4 mL of DMAC was degassed and purged with N2 for 3 times, and then the mixture was stirred at 130°C for 12 hours under N2 atmosphere. Then it was partitioned between 10 mL of water and 30 mL of ethyl acetate. The organic phase was separated, washed with 30 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =10: 1 to 5: 1) to give 140 mg of compound 2 (944.9 pmol, 95.5% yield) as a yellow oil.

Preparation of compound 3:

To a solution of 3-amino-5-methoxybenzonitrile (140 mg, 944.9 pmol, 1 eq) in 5 mL of dichloromethane was added BBr3 (1.4 g, 5.7 mmol, 6 eq) at 0°C. The mixture was stirred at

15°C for 1 hour. Then it was partitioned between 10 mL of saturated NaHCCb and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 200 mg of crude compound 3 as a yellow solid, which was used into the next step directly.

Preparation of compound 4:

To a solution of 3-amino-5-hydroxybenzonitrile (200 mg, 1.5 mmol, 1 eq) and KOH (836.6 mg, 14.9 mmol, 10 eq) in 3 mL of acetonitrile and 3 mL of water was added dropwise 1- [[bromo(difluoro)methyl]-ethoxy-phosphoryl]oxye thane (477.7 mg, 1.8 mmol, 1.2 eq) at -78°C. After addition, the resulting mixture was stirred at 20°C for 2 hours. Then it was partitioned between 10 mL of water and 35 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC(Si02, eluting with petroleum ether:ethyl acetate =1 : 1) to give 96 mg of compound 4 (521.3 pmol, 34.9% yield) as a yellow oil. Then compound 4 was used into an amide coupling reaction by using a normal method to give

239.

compound 245:

J H NMR (400 MHz, DMSO-de) d ppm 10.95 (s, 1 H) 8.18 (s, 1 H) 8.11 (s, 1 H) 7.74 (s, 1 H) 7.59 (d, J = 8.50 Hz, 1 H) 7.33 - 7.40 (m, 2 H) 3.79 (br t, / = 6.38 Hz, 2 H) 3.57 (t, J = 7.32 Hz,

2 H) 3.00 (br t, / = 7.07 Hz, 2 H) 2.40 - 2.45 (m, 2 H) 1.63 (sxt, J = 7.25 Hz, 2 H) 0.93 (t, J = 7.38 Hz, 2 H)

LCMS (ESI+): m/z 455.0 (M+H) Synthesis of the aniline of 245

Experimental Procedures:

Preparation of compound 2:

To a solution of l,3-dibromo-5-chloro-benzene (3 g, 11.1 mmol, 1 eq) and cyclobutanone (1.2 g, 16.7 mmol, 1.2 mL, 1.5 eq) in 30 mL of tetrahydrofuran was added dropwise n-BuLi (2.5 M, 6.7 mL, 1.5 eq) at -78°C under N2. The mixture was stirred at -78°C for 1 hour. Then it was partitioned between 100 mL of sat.NPLCl and 150 mL of ethyl acetate. The organic phase was separated, washed with 100 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate = 100: 1 to 50: 1) to give 1.7 g of compound 2 (6.5 mmol, 58.6% yield) as a yellow oil.

Preparation of compound 4:

To a solution of l-(3-bromo-5-chloro-phenyl)cyclobutanol (0.7 g, 2.7 mmol, 1 eq), BINAP (166.7 mg, 267.6 pmol, 0.1 eq), Pd2(dba)3 (24.5 mg, 26.8 pmol, 0.01 eq) and t-BuOK (750.8 mg, 6.7 mmol, 2.5 eq ) in 10 mL of toluene was added diphenylmethanimine (582.1 mg, 3.2 mmol, 539.0 pL, 1.2 eq). The mixture was stirred at 110°C for 1 hour. Then it was partitioned between 20 mL of water and 40 mL of ethyl acetate. The organic phase was separated, washed with 30 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (SiCh, eluting with a gradient of petroleum ether : ethyl acetate = 100: 1 to 20: 1) to give 0.9 g of crude compound 4 as a yellow oil, which was used directly into the next step without purification.

Preparation of compound 5:

4 5

A mixture of l-[3-(benzhydrylideneamino)-5-chloro-phenyl]cyclobutanol (0.9 g, 2.5 mmol, 1 eq) in HC1 (6 M, 9.0 mL, 21.7 eq) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 20°C for 12 hours under N2 atmosphere. Then it was poured into 3 mL of ice-water, adjusted pH~10 by using IN NaOH, and extracted twice with 50 mL of ethyl acetate. The organic phase was separated, washed with 30 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (TFA condition) to give product. Then it was partitioned between 2 mL of sat. NaHCCL and 4 mL of ethyl acetate. The organic phase was separated, washed with 3 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 55 mg of compound 5 (278.3 pmol, 11.2% yield) as a yellow oil.

Then compound 5 was used into an amide coupling reaction by using a normal method to give

245. EXAMPLE 50: Synthesis of compound 217 (large scale reaction)

Experimental procedures:

Preparation of compound 2:

To a solution of compound 1 (50 g, 242.2 mmol, 1 eq ) in 1 L of toluene and 100 mL of H2O was added potassium cyclopropyl(trifluoro)boranuide (39.4 g, 266.4 mmol, 1.1 eq), t- BuOK (81.5 g, 726.5 mmol, 3 eq) and Pd(dppf)Cl2.CH2Cl2 (19.8 g, 24.2 mmol, 0.1 eq) and purged with N2 for 3 times. The mixture was stirred at 90°C for 16 hours. Then it was filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =1:0 to 3:1) to get 37.5 g of compound 2 (223.7 mmol, 92.4% yield) as a yellow oil.

Preparation of 217:

A solution of compound 3 (32.9 g, 119.3 mmol, 1 eq) in 100 mL of dimethyl form amide was added TEA (36.2 g, 357.9 mmol, 49.8 mL, 3 eq) at 0°C to get a mixture. A solution of HATU (49.9 g, 131.2 mmol, 1.1 eq) in 150 mL of dimethyl formamide was added dropwise to above mixture at 0°C. The mixture was stirred at 20°C for 0.5 hour. A solution of compound 2 (20 g, 119.3 mmol, 1 eq ) in 50 mL of dimethyl form amide was added dropwise to the mixture at 20°C. Then it was stirred at 20°C for 12 hours. After reaction, it was partitioned between 800 mL of water and 1000 mL of methyl-tert-butyl ether. The organic phase was separated, washed with 200 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (Si(¾, eluting with a gradient of petroleum ether : ethyl acetate = 1 :0 to 1: 1).

The obtained crude product was further purified by recrystallization in ethyl acetate and methanol: It was added into 350 mL of methanol at 80°C to give a suspension. 100 mL of ethyl acetate was added to the mixture with stirring for 0.5 hour to give a light yellow solution. Then the mixture was cooled naturally to 20°C over 12 hours. The mixture was filtered, and the filter cake was dried in vacuum at 50°C, then it was added 1000 mL of deionized water and stirred at 100°C for 2 hours. The resulting mixture was filtered to get a filter cake, which was added 1000 mL of deionized water and stirred at 100°C for another 0.5 hour. Then it was filtered to get a filter cake. The filter cake was dried in drying oven at 60°C to get 25.7 g of 217(60.4 mmol, 50.6% yield, 100.0% purity) as a white solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.60 - 10.66 (m, 1 H) 7.64 - 7.67 (m, 1 H) 7.55 - 7.59 (m, 1 H) 7.29 - 7.37 (m, 3 H) 6.89 - 6.92 (m, 1 H) 3.73 - 3.82 (m, 2 H) 3.52 - 3.60 (m, 2 H) 2.38 - 2.46 (m, 2 H) 1.89 - 1.97 (m, 1 H) 0.94 - 1.02 (m, 2 H) 0.65 - 0.71 (m, 2 H)

LCMS (ESI+): m/z 425.0 (M+H)

Compounds 290, 291 were prepared analogously to 217 coupling the appropriate acid with the aniline 2:

compound 290:

J H NMR (400 MHz, DMSO-de) d ppm 10.68 - 10.77 (m, 1 H), 7.60 - 7.66 (m, 1 H), 7.50 - 7.56

(m, 1 H), 7.26 - 7.35 (m, 2 H), 6.90 - 6.94 (m, 1 H), 3.77 - 3.84 (m, 2 H), 3.57 - 3.64 (m, 2 H),

2.43 (br t, / = 6.88 Hz, 2 H), 1.89 - 1.98 (m, 1 H), 0.93 - 1.03 (m, 2 H), 0.64 - 0.72 (m, 2 H).

LCMS (ESI+): m/z 459.0 (M+H) compound 291:

J H NMR (400 MHz, DMSO-de) d ppm 10.70 - 10.77 (m, 1 H) 7.61 - 7.66 (m, 1 H) 7.33 (br dd, J = 11.32, 1.81 Hz, 1 H) 7.30 (s, 1 H) 7.19 - 7.23 (m, 1 H) 6.90 - 6.94 (m, 1 H) 3.77 - 3.83 (m, 2 H) 3.57 - 3.65 (m, 2 H) 2.38 - 2.46 (m, 2 H) 1.88 - 1.99 (m, 1 H) 0.93 - 1.02 (m, 2 H) 0.64 - 0.73 (m, 2 H)

LCMS (ESI+): m/z 443.0 (M+H)

EXAMPLE 51: Synthesis of Compound 304

compound 304: J H NMR (400 MHz, DMSO-de) d ppm 10.61 - 10.72 (m, 1 H) 8.81 - 8.84 (m, 1 H) 8.71 - 8.74 (m, 1 H) 8.60 - 8.64 (m, 2 H) 7.56 - 7.60 (m, 1 H) 7.47 - 7.51 (m, 1 H) 7.40 - 7.44 (m, 1 H) 7.34 - 7.37 (m, 1 H) 7.31 - 7.32 (m, 1 H) 6.81 - 6.87 (m, 1 H) 3.75 - 3.82 (m, 2 H) 3.53 - 3.58 (m, 2 H) 2.39 - 2.45 (m, 2 H)

LCMS (ESI+): m/z 478.0 (M+H)

Synthesis of the aniline of 304

Experimental procedures:

Preparation of compound 2:

To a solution of compound 1 (200 mg, 652.4 pmol, 1 eq) in 3 mL of toluene was added pyrimidin-5-amine (124.1 mg, 1.3 mmol, 2 eq), Xantphos (56.6 mg, 97.8 pmol, 0.2 eq), t-BuOK (146.4 mg, 1.3 mmol, 2 eq) and Pd 2 (dba) 3 (59.7 mg, 65.2 pmol, 0.1 eq) and purged with N2for 3 times. The mixture was stirred at 110°C for 12 hours. Then it was filtered and concentrated under reduced pressure to give the crude product, which was purified by column

chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =1:0 to 1:1) to get 100 mg of compound 2 (311.8 pmol, 47.8% yield) as a yellow oil.

Preparation of compound 3:

To a solution of compound 2 (100 mg, 311.8 pmol, 1 eq ) in 3 mL of dichloromethane was added 1 mL of TFA. The mixture was stirred at 25 °C for 0.5 hour. Then it was adjusted pH to 8 by using sat.NaHCCb, partitioned between 5 mL of water and 30 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to get 65 mg of crude compound 3 as a yellow oil, which was used directly into the next step without purification.

Then compound 3 was used into an amide coupling reaction by using a normal method to give

304. EXAMPLE 52: Synthesiws of Compound 262

Compound 262:

J H NMR (400 MHz, DMSO-de) d = 10.72 - 10.67 (m, 1H), 7.73 - 7.70 (m, 1H), 7.60 - 7.56 (m, 1H), 7.55 - 7.51 (m, 1H), 7.39 - 7.31 (m, 2H), 7.11 - 7.07 (m, 1H), 4.00 - 3.91 (m, 1H), 3.83 - 3.75 (m, 2H), 3.62 - 3.54 (m, 2H), 3.53 - 3.42 (m, 2H), 2.87 - 2.77 (m, 1H), 2.46 - 2.40 (m, 2H),

1.83 - 1.74 (m, 1H), 1.74 - 1.65 (m, 1H), 1.61 - 1.48 (m, 1H), 1.34 - 1.22 (m, 1H), 1.16 - 1.09 (m, 3H)

LCMS (ESI+): m/z 483.1 (M+H)

Synthesis of the aniline of 262

Experimental Procedures:

Preparation of compound 3:

1 3 LiHMDS (1 M, 2.6 mL, 1.5 eq ) was added to a solution of compound 1 (200 mg, 1.7 mmol, 1 eq) in 20 mL of terahydrofuran at -70°C under N2. The mixture was warmed to -40°C and stirred for 0.5 hour. Then a solution of compound 2 (751.2 mg, 2.1 mmol, 1.2 eq) in 20 mL of terahydrofuran was added dropwise to the mixture at -40°C. The mixture was stirred at -40°C for 0.1 hour. Then it was warmed to -20°C and stirred for 0.9 hour. After reaction, it was warmed to 0°C and quenched by addition of 10 mL of water. Then it was partitioned between 5 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate=l:0 to 20: 1) to give 120 mg of compound 3 (487.4 pmol, 27.82% yield) as a white oil.

Preparation of compound 4:

To a solution of compound 4A (5 g, 24.2 mmol, 1 eq), compound B (6.8 g, 26.6 mmol,

1.1 eq) and NaOAc (3.9 g, 48.4 mmol, 2 eq) in 100 mL of dioxane was added Pd(dppf)Cl2 (1.8 g, 2.4 mmol, 0.1 eq) under N2. The mixture was stirred at 110°C for 12 hours. Then it was partitioned between 100 mL of water and 150 mL of ethyl acetate. The organic phase was separated, washed with 100 mL of brine, dried over Na2SC>4, filtered and concentrated in vacuum. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate=20: l to 3: 1) to give 4 g compound 4 (15.8 mmol, 65.2% yield) as a yellow oil.

Preparation of compound 5:

To a solution of compound 4 (80 mg, 315.5 pmol, 1 eq), compound 3 (116.5 mg, 473.3 pmol, 1.5 eq) and CS2CO3 (308.4 mg, 946.6 pmol, 3 eq) in 2 mL of dioxane and 0.4 mL of water was added Pd(dppf)Cl2 (23.1 mg, 31.6 pmol, 0.1 eq) under N2. The mixture was stirred at 100°C for 12 hours. Then it was partitioned between 5 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5mL of brine, dried over Na2SC>4, filtered and concentrated in vacuum. The residue was purified by prep- TLC (S1O2, eluting with petroleum ether : ethyl acetate=3: l) to give 40 mg of compound 5 (178.8 pmol, 56.7% yield) as a yellow oil.

Preparation of compound 6:

To a solution of compound 5 (40 mg, 178.8 pmol, 1 eq) in 2 mL of ethyl acetate was added Pd/C (20 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with ¾ several times. The mixture was stirred at 20°C for 1 hour under ¾ (15psi) atmosphere. After reaction, it was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by prep- HPLC (TFA condition) to give 20 mg of compound 6 (58.9 pmol, 32.9% yield, TFA) as a brown solid.

Then compound 6 was used into an acid amide coupling reaction by using a normal method to give 262.

263 was made analogously to 262.

J H NMR (400 MHz, DMSO-de) d = 10.71 - 10.67 (m, 1H), 7.73 - 7.69 (m, 1H), 7.60 - 7.52 (m, 2H), 7.39 - 7.31 (m, 2H), 7.11 - 7.08 (m, 1H), 3.82 - 3.76 (m, 2H), 3.72 - 3.67 (m, 2H), 3.60 - 3.54 (m, 2H), 3.04 - 2.90 (m, 1H), 2.47 (br s, 2H), 1.72 - 1.63 (m, 2H), 1.56 - 1.47 (m, 1H), 1.47 - 1.37 (m, 1H), 1.28 - 1.22 (m, 3H), 1.20 - 1.14 (m, 3H)

LCMS (ESI+): m/z 497.1 (M+H)

EXAMPLE 53: Synthesis of Compound 243

compound 243:

J H NMR (400 MHz, DMSO-de) d ppm 10.74 - 10.71 (m, 1H), 7.78 - 7.75 (m, 1H), 7.60 - 7.55 (m, 2H), 7.38 - 7.30 (m, 2H), 7.12 - 7.09 (m, 1H), 4.84 - 4.78 (m, 1H), 4.01 - 3.94 (m, 1H), 3.85 - 3.81 (m, 1H), 3.80 - 3.76 (m, 2H), 3.60 - 3.53 (m, 2H), 2.46 - 2.38 (m, 2H), 2.37 - 2.27 (m, 1H), 1.97 - 1.88 (m, 2H), 1.70 - 1.58 (m, 1H)

LCMS (ESI+): m/z 455.0(M+H)

Synthesis of the aniline of 243

Experimental Procedures:

Preparation of compound 2A, 2B:

To a solution of 3-bromo-5-chloro-aniline (1 g, 4.8 mmol, 1 eq), 2,3-dihydrofuran (678.9 mg, 9.7 mmol, 732.4 pL, 2 eq), PPI13 (254.1 mg, 968.7 pmol, 0.2 eq) and K2CO3 (2.0 g, 14.5 mmol, 3 eq) in 5 mL of N,N- dimethylformamide was added Pd(OAc)2 (108.7 mg, 484.3 pmol, 0.1 eq) at 20°C under N2. The mixture was stirred at 110°C for 4 hours. After reaction, it was diluted with 10 mL of ethyl acetate and 10 mL of water. The organic layer was separated and concentrated under reduced pressure to get a residue. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ethenethyl acetate=20:l to 5:1) to give 180 mg of compound 2A (920.0 pmol, 19.0% yield) and 300 mg of compound 2B (1.5 mmol, 31.7% yield) as all yellow oil.

Preparation of compound 3:

To a solution of compound 2A(180 mg, 920.0 pmol, 1 eq) in 5 mL of ethyl acetate was added Pd/C (50 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with ¾ several times. The mixture was stirred at 20°C for 0.5 hour under ¾ (15PSI) atmosphere. After reaction, it was filtered and the filtrate was concentrated under reduced pressure to give 90 mg of crude compound 3 as a yellow oil, which was used directly into the next step without purification.

Compound 2B can be also hydrogenated analogously to give compound 3.

Then compound 3 was used into an acid amide coupling reaction by using a normal method to give 243.

EXAMPLE 54: Synthesis of compounds 267 and 268

243 (240 mg, 527.1 pmol) was separated by SFC (column :DAICEL CHIRALPAK

AD(250mm*30mm,10pm); mobile phase: [Neu-MeOH];B%: 50%-50%,18min) to give 43.6 mg of 267 (95.7 m ihoI, 18.2% yield, 100% purity) and 66.7 mg of 268 (144.9 m ihoI, 27.5% yield, 98.9% purity) as white solid.

The structures of the two isomers were random assignment.

J H NMR (400 MHz, DMSO-de) d ppm 10.77 - 10.73 (m, 1H), 7.79 - 7.75 (m, 1H), 7.62 - 7.55 (m, 2H), 7.39 - 7.30 (m, 2H), 7.13 - 7.09 (m, 1H), 4.86 - 4.78 (m, 1H), 4.02 - 3.94 (m, 1H), 3.86

- 3.75 (m, 3H), 3.61 - 3.54 (m, 2H), 2.48 - 2.39 (m, 2H), 2.38 - 2.28 (m, 1H), 1.98 - 1.88 (m, 2H), 1.70 - 1.59 (m, 1H)

LCMS (ESI+): m/z 455.0 (M+H)

268:

J H NMR (400 MHz, DMSO-de) d ppm 10.77 - 10.73 (m, 1H), 7.79 - 7.75 (m, 1H), 7.62 - 7.55 (m, 2H), 7.39 - 7.31 (m, 2H), 7.13 - 7.09 (m, 1H), 4.87 - 4.78 (m, 1H), 4.03 - 3.94 (m, 1H), 3.81 (br s, 1H), 3.81 - 3.75 (m, 2H), 3.63 - 3.52 (m, 2H), 2.46 - 2.40 (m, 2H), 2.37 - 2.28 (m, 1H), 2.01 - 1.87 (m, 2H), 1.72 - 1.58 (m, 1H) LCMS (ESI+): m/z 455.0 (M+H)

Usual amide coupling method used to make compound 218:

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 8.30 (s, 1 H) 8.14 - 8.19 (m, 1 H) 7.97 - 8.02 (m, 1 H) 7.80 - 7.85 (m, 1 H) 7.53 (d, / = 1.54 Hz, 1 H) 7.44 - 7.47 (m, 2 H) 3.92 - 3.96 (m, 3 H) 3.77 - 3.84 (m, 2 H) 3.39 - 3.46 (m, 2 H) 2.53 - 2.63 (m, 2 H) LCMS (ESI+): m/z 443.0 (M+H)

EXAMPLE 55: Synthesis of Compound 302

compound 302: J H NMR (400MHz, DMSO-de) d ppm 10.75 (s, 1 H), 7.81 (s, 1 H), 7.60 (br s, 1 H), 7.56 (br d, J = 8.8 Hz, 1 H), 7.42 (s, 1 H), 7.35 (br d, / = 8.8 Hz, 1 H), 7.29 (br s, 2 H), 6.81 (br s, 1 H), 3.82 (s, 3 H), 3.77 (br t, / = 6.3 Hz, 2 H), 3.56 (br t, / = 7.3 Hz, 2 H), 2.44 - 2.39 (m, 2 H), 2.07 (s, 1 H)

LCMS (ESI+): m/z 481.0 (M+H)

Synthesis of the aniline of compound 302:

Experimental Procedures:

Preparation of compound 2:

To a mixture of l-methylpyrazol-4-ol (111.8 mg, 1.1 mmol, 1 eq) in 2 mL of N,N- Dimethylformamid was added NaH (68.4 mg, 1.7 mmol, 60% purity, 1.5 eq) in 3 portions at 0°C. The mixture was stirred for 0.5 hour and then l-chloro-3-fluoro- 5-nitro-benzene (200 mg, 1.1 mmol, 1 eq) was added. The mixture was allowed to warmed to 20°C and stirred for 1 hour. After reaction, it was diluted with 20 mL of water and 20 mL of ethyl acetate. The organic layer was separated and concentrated under reduced pressure to give 250 mg of crude compound 2 as a light yellow oil, which was used directly into the next step without purification.

Preparation of compound 3:

The mixture of compound 2 (250 mg, 985.6 pmol, 1 eq), Fe (275.2 mg, 4.9 mmol, 5 eq ) and NH4CI (263.6 mg, 4.9 mmol, 5 eq) in 5 mL of EtOH and 1 mL of water was stirred for 12 hours at 70°C. After reaction, it was filtered and the filtrate was concentrated under reduced pressure to get a residue, which was dissolved in 20 mL of ethyl acetate and 20 mL of water. The organic layer was separated, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 150 mg of compound 3 as a light yellow oil, which was used directly into the next step without purification.

Then compound 3 was used into an amide coupling reaction by using a normal method to give

302.

EXAMPLE 56:: Synthesis of compound 211:

Experimental Procedures:

Preparation of compound 1:

To a solution of 3-bromo-5-chloro-aniline (3.6 g, 17.3 mmol, 1 eq) and methyl 2-chloro- 5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoate (5 g, 17.3 mmol, 1 eq) in 50 mL of Tol. was added AlMe3 (2 M, 12.9 mL, 1.5 eq) at 0°C. The mixture was stirred at 110°C for 3hrs. Then it was partitioned between 30 mL of water and 100 mL of ethyl acetate. The organic phase was separated, washed with 20 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =10: 1 to 1 :5) to give 4.2 g of compound 1 (9.1 mmol, 52.4% yield) as a yellow solid.

Preparation of compound 2:

To a solution of compound 1 (100 mg, 215.4 pmol, 1 eq), 2-(3,4-dihydro-2H-pyran-6- yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (54.3 mg, 258.5 pmol, 1.2 eq) and K3PO4 (91.5 mg, 430.9 pmol, 2 eq) in 2 mL of tetrahydrofuran and 0.5 mL of water was added tBuXPhos Pd G3 (17.1 mg, 21.5 pmol, 0.1 eq) and purged with N 2 for3 times. The mixture was stirred at 80°C for 15 hours. Then it was filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =1 : 1) to get 40 mg of compound 2 (85.6 pmol, 39.7% yield) as a white solid.

Preparation of 211:

A mixture of compound 2 (20 mg, 42.8 pmol, 1 eq), Pd/C (20 mg, 10% purity) in 1 mL of ethyl acetate was degassed and purged with ¾ for 5 times, and then the mixture was stirred at 25°C for 10 min under ¾ (15 psi) atmosphere. Then it was filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (TFA condition) to get 3.4 mg of compound 211 (7.12 pmol, 16.6% yield, 98.3% purity) as a white solid.

J H NMR (400 MHz, CHLOROF ORM-d) d ppm 8.01 - 8.06 (m, 1 H) 7.73 - 7.75 (m, 1 H) 7.49 - 7.52 (m, 1 H) 7.47 - 7.49 (m, 1 H) 7.44 - 7.46 (m, 1 H) 7.41 - 7.43 (m, 1 H) 7.16 - 7.18 (m, 1 H) 4.28 - 4.35 (m, 1 H) 4.11 - 4.18 (m, 1 H) 3.78 - 3.85 (m, 2 H) 3.55 - 3.66 (m, 1 H) 3.39 - 3.46 (m, 2 H) 2.53 - 2.64 (m, 2 H) 1.92 - 1.99 (m, 1 H) 1.83 - 1.90 (m, 1 H) 1.61 - 1.74 (m, 4 H)

LCMS (ESI+): m/z 469.1 (M+H)

Conpound 209 was prepared through a normal Suzuki reaction method by using the appropriate boronic acid with aryl bromide 1.

compound 209 :

J H NMR (400 MHz, DMSO-de) d ppm 10.94 (s, 1 H) 9.24 (s, 1 H) 9.11 (s, 2 H) 7.98 (t, / = 1.77 Hz, 1 H) 7.95 (t, / = 1.65 Hz, 1 H) 7.69 (t, / = 1.71 Hz, 1 H) 7.57 - 7.63 (m, 1 H) 7.34 - 7.40 (m, 2 H) 3.79 (t, J = 6.48 Hz, 2 H) 3.57 (t, / = 7.34 Hz, 2 H) 2.43 (t, J = 7.09 Hz, 2 H)

LCMS (ESI+): m/z 463.0 (M+H) EXAMPLE 57: Synthesis of compound 233:

Experimental Procedures:

Preparation of compound 2:

To a solution of compound 1 (300 mg, 646.3 pmol, 1 eq), 2-isopropenyl-4, 4,5,5- tetramethyl-l,3,2-dioxaborolane (119.5 mg, 711.0 pmol, 1.1 eq) and K2CO3 (268.0 mg, 1.9 mmol, 3 eq) in 1 mL of water and 4 mL of dioxane was added Pd(dppf)Cl2 (47.3 mg, 64.6 pmol, 0.1 eq) under N2. The mixture was purged with N2 for3 times. Then the mixture was heated to

110°C and stirred for 12 hours. The mixture was cooled to 20°C and partitioned between 15 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chouromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate = 10: 1 to 5:2) to give 190 mg of compound 2 (446.7 pmol, 69.1% yield) as a white solid. Preparation of compound 233:

To a solution of ZnEt2 (1 M, 3.1 mL, 10 eq) in 2 mL of dichloromethane was added TFA (348.5 mg, 3.1 mmol, 226.3 mT, 10 eq) dropwise at 0°C. The mixture was stirred at 0°C for 0.3 hour. CH2I2 (818.6 mg, 3.1 mmol, 246.6 pL, 10 eq) was added dropwise to the mi ture at 0°C and stirred for 0.3 hour. Then a solution of compound 2 (130 mg, 305.7 pmol, 1 eq) in 2 mL of dichloromethane was added dropwise to the mixture at 0°C. The mixture was warmed to 25 °C and stirred at 25 °C for 1 hour. After reaction, it was partitioned between 5 mL of water and 5 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to get a residue. The residue was purified by prep- HPLC (TFA condition) to get 19.7 mg of 233 (44.8 pmol, 14.7% yield, 100% purity) as a white solid.

J H NMR (400MHz, DMSO-de) d ppm 10.67 (s, 1 H), 7.72 (s, 1 H), 7.57 (d, / = 8.8 Hz, 1 H), 7.48 (s, 1 H), 7.40 - 7.29 (m, 2 H), 6.99 (s, 1 H), 3.78 (t, / = 6.5 Hz, 2 H), 3.56 (t, / = 7.4 Hz, 2 H), 3.33 (s, 2 H), 2.46 - 2.39 (m, 2 H), 1.36 (s, 3 H), 0.89 - 0.73 (m, 4 H)

LCMS (ESI+): m/z 439.0 (M+H)

EXAMPLE 58: Synthesis of compound 212:

Experimental Procedures:

Preparation of compound 3:

1 3

To the solution of compound 1 (1 g, 2.2 mmol, 1 eq ), compound 2 (656.5 mg, 2.6 mmol, 1.2 eq) and AcONa (530.2 mg, 6.5 mmol, 3 eq) in 10 mL of dioxane and 1 mL of water was added Pd(dppf)Cl2 (157.6 mg, 215.4 pmol, 0.1 eq) under N2 at 20°C. The mixture was degassed under vacuum and purged with N2 5 times. Then the mixture was heated to 70°C and stirred for 12 hours. After reaction, it was cooled to 20°C and poured into 20 mL of water. The aqueous phase was extracted with 10 mL of ethyl acetate, separated and the organic layer was dried over Na2SC>4, filtered and concentrated under reduced pressure to get crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate=10: l to 1 :1) to give 400 mg of compound 3 ( 932.2 pmol, 43.3% yield) as a light yellow solid. Preparation of compound 212:

Pd(dppf)Cl2 (13.6 mg, 18.6 pmol, 0.1 eq ) was added to a mixture of compound 3 (80 mg, 186.4 pmol, 1 eq), 4-bromo- 2-methyl-pyridine (32.1 mg, 186.4 pmol, 1 eq) and K2CO3 (77.3 mg, 559.3 pmol, 3 eq) in 2 mL of dioxane and 0.2 mL of water at 20°C. Then the mixture was purged with N2 for 3 times. The mixture was heated to 100°C and stirred for 12 hours. After reaction, it was diluted with 20 mL of ethyl acetate. Then 5 g of silica gel was added to the mixture and the mixture was stirred for 10 mins. The mixture was filtered and the filtrate was concentrated under reduced pressure to get residue, which was purified by prep- HPLC (TFA condition) to give 24.1 mg of 212 (40.6 pmol, 21.8% yield, 99.4% purity, TFA salt) as a white solid.

J H NMR (400MHz, DMSO-de) d ppm 10.99 (s, 1 H), 8.77 (d, J = 5.9 Hz, 1 H), 8.18 (s, 1 H), 8.05 (s, 1 H), 7.96 (t, / = 1.7 Hz, 1 H), 7.93 (br d, / = 5.4 Hz, 1 H), 7.77 (t, / = 1.7 Hz, 1 H), 7.63 - 7.58 (m, 1 H), 7.40 - 7.34 (m, 1 H), 7.40 - 7.34 (m, 1 H), 3.79 (t, / = 6.5 Hz, 2 H), 3.57 (t, J =7.4 Hz, 2 H), 2.70 (s, 3 H), 2.43 (t, J = 7.0 Hz, 2 H).

LCMS (ESI+): m/z 476.0 (M+H)

213 was prepared analogously coupling the appropriate aryl bromide with boronic acid 3.

J H NMR (400MHz, DMSO-de) d ppm 10.87 (s, 1 H), 8.02 - 7.84 (m, 2 H), 7.59 (d, / = 9.0 Hz, 1 H), 7.54 - 7.47 (m, 2 H), 7.41 - 7.33 (m, 2 H), 6.56 (d, / = 1.5 Hz, 1 H), 6.46 (dd, / = 1.8, 6.8 Hz, 1 H), 3.79 (t, J =6.5 Hz, 2 H), 3.57 (t, J = 7.3 Hz, 2 H), 2.46 - 2.40 (m, 2 H). LCMS (ESI+): m/z 478.0 (M+H)

216 was prepared analogously coupling the appropriate borate with aryl bromide.

compound 216:

J H NMR (400 MHz, DMSO-de) d ppm 10.81 - 10.92 (m, 1 H) 7.94 - 7.97 (m, 1 H) 7.89 - 7.93 (m, 1 H) 7.79 - 7.84 (m, 1 H) 7.58 - 7.62 (m, 1 H) 7.52 - 7.56 (m, 1 H) 7.34 - 7.40 (m, 2 H) 6.62

- 6.65 (m, 1 H) 6.50 - 6.55 (m, 1 H) 3.73 - 3.84 (m, 2 H) 3.53 - 3.63 (m, 2 H) 3.42 - 3.49 (m, 3 H) 2.40 - 2.44 (m, 1 H) 2.39 - 2.45 (m, 1 H)

LCMS (ESI+): m/z 492.0 (M+H)

Synthesis of corresponding borate:

To a solution of compound 1 (1 g, 2.2 mmol, 1 eq ), 4,4,5,5-tetramethyl-2-(4,4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (601.8 mg, 2.4 mmol, 1.1 eq) and AcOK (422.9 mg, 4.3 mmol, 2 eq) in 20 mL of dioxane was added Pd(dppf)Cl2 (157.6 mg, 215.4 umol, 0.1 eq) and purged with N2 for 3 times. The mixture was stirred at 100°C for 12 hours. Then it was filtered, partitioned between 25 mL of water and 50 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate = 1 :0 to 0: 1) to get 1.1 g of compound 2 (2.2 mmol, 99.9% yield) as a yellow oil. Then compound 2 was used into a suzuki reaction by using a normal method to give 216. EXAMPLE 59: Synthesis of compound 306:

Experimental Procedures:

Preparation of compound 3:

K2CO3 (1.1 g, 8.1 mmol, 3 eq) was added to a solution of tert-butyl 4-hydroxypyrazole- 1-carboxylate (500 mg, 2.7 mmol, 1 eq) and l-chloro-3- fluoro-5-nitro-benzene (476.5 mg, 2.7 mmol, 1 eq) in 2 mL of DMF under N2. The mixture was heated to 90°C and stirred for 1 hour. The mixture was partitioned between 20 mL of ethyl acetate and 20 mL of water. The organic layer was separated and concentrated under reduced pressure to get a residue. The residue was purified by column chouromatography (Si(¾, eluting with a gradient of petroleum ether : Ethyl acetate = 1 :0 to 10:1) to give 700 mg of compound 3 (2.1 mmol, 75.9% yield) as a colorless oil.

Preparation of compound 4:

TFA (1.5 g, 13.5 mmol, 1 mL, 15.3 eq) was added to a solution of compound 3 (300 mg, 883.1 pmol, 1 eq) in 2 mL of dichloromethane at 25°C. The mixture was stirred at 25°C for 2 hours. The mixture was concentrated under reduced pressure to give a residue. The residue was partitioned between 10 mL of dichloromethane and 10 mL of saturated solution of NaHCCb. The organic layer was separated, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 200 mg of crude compound 4 as a light yellow oil, which was used directly into the next step without purification.

Preparation of compound 5:

NaH (66.8 mg, 1.7 mmol, 60% purity, 2 eq) was added to a solution of compound 4 (200 mg, 834.7 pmol, 1 eq) in 2 mL of THF at 0°C. The mixture was warmed to 20°C and stirred for 1 hour. A solution of SEM-C1 (167.0 mg, 1.0 mmol, 177.3 pL, 1.2 eq) in 1 mL of THF was added dropwise to the mixture at 20°C. The mixture was stirred for 1 hour. After reaction, it was concentrated under reduced pressure to remove solvent. The residue was partitioned between 5 mL of ethyl acetate and 5 mL of water. The organic layer was separated and concentrated under reduced pressure to give 250 mg of crude compound 5 as an off-white solid, which was used directly into the next step without purification. Preparation of compound 6:

The mixture of compound 5 (250 mg, 675.9 pmol, 1 eq), Fe (188.7 mg, 3.4 mmol, 5 eq) and NH4CI (180.8 mg, 3.4 mmol, 5 eq) in 3 mL of ethanol and 1 mL of water was heated to 70°C and stirred for 1 hour. The mixture was filtered and the filtrate was concentrated under reduced pressure to get a crude product, which was partitioned between 5 mL of water and 5 mL of ethyl acetate. The organic layer was separated and dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 150 mg of crude compound 6 as a light yellow oil, which was used directly into the next step without purification.

Preparation of compound 8:

HATU (82.8 mg, 217.6 pmol, 1.2 eq) was added to a solution of compound 6 (61.6 mg, 181.4 pmol, 1 eq), compound 7 (50 mg, 181.4 pmol, 1 eq) and TEA (36.7 mg, 362.7 pmol, 2 eq) in 2 mL of dimethyl formamide at 0°C. The mixture was warmed to 25°C and stirred for 12 hours. The mixture was partitioned between 10 mL of ethyl acetate and 10 mL of water. The organic layer was separated and concentrated under reduced pressure to give 100 mg of crude compound 8 as a light yellow oil, which was used directly into the next step without purification. Preparation of compound 306:

TFA (1.5 g, 13.5 mmol, 1 mL, 80.7 eq ) was added to a solution of compound 8 (100 mg, 167.3 m ihoI, 1 eq) in 1 mL of dichloromethane at 25°C. The mixture was stirred for 2 hours. After reaction, it was concentrated under reduced pressure to get a residue. The residue was purified by prep- HPLC (TFA condition) to give 22 mg of compound 306 (37.8 pmol, 22.6% yield, 99.9% purity, TFA salt) as a white solid.

J H NMR (400MHz, DMSO-de) d ppm 10.75 (s, 1 H), 7.66 (br s, 2 H), 7.60 (t, / = 1.8 Hz, 1 H), 7.56 (d, / = 8.8 Hz, 1 H), 7.35 (dd, / = 2.9, 8.8 Hz, 1 H), 7.31 - 7.25 (m, 2 H), 6.80 (t, / = 2.1 Hz, 1 H), 3.77 (t, J = 6.4 Hz, 2 H), 3.55 (t, J = 7.4 Hz, 2 H), 2.41 (quin, J = 6.9 Hz, 2 H)

LCMS (ESI+): m/z 467.0 (M+H)

Example 60: Synthesis of Compound 231

Compound 231 was prepared through three methods:

Method A: (batch 1)

Experimental Procedures:

Preparation of compound 3:

To a solution of 3-chloro-5-nitro-phenol (50 mg, 288.1 pmol, 1 eq) in 2 mL of acetonitrile was added K2CO3 (79.6 mg, 576.2 pmol, 2 eq) and 4-bromotetrahydropyran (142.6 mg, 864.3 pmol, 3 eq). The mixture was stirred at 80°C for 36 hrs. Then KI (47.8 mg, 288.1 pmol, 1 eq) was added and stirred at 80°C for another 12 hrs. After reaction, it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =5: 1) to give 30 mg of compound 3 (116.4 pmol, 40.4% yield) as a yellow oil.

Preparation of compound 4:

To a solution of 4-(3-chloro-5-nitro-phenoxy)tetrahydropyran (30 mg, 116.4 pmol, 1 eq) in 2 mL of ethyl alcohol and 1 mL of water was added Fe (32.5 mg, 582.1 pmol, 5 eq) and NH4CI (31.1 mg, 582.14 pmol, 5 eq). The mixture was stirred at 85°C for 12 hours. After reaction, it was filtered, the filtrate was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 30 mg of crude compound 4 as a yellow oil, which was used into the next step directly. Preparation of compound 231 (batch 1):

4 231

To a solution of 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoic acid (43.6 mg, 158.1 pmol, 1.2 eq ) in 2 mL of dimethyl form amide was added 3-chloro-5-tetrahydropyran-4-yloxy- aniline (30 mg, 131.8 pmol, 1 eq), TEA (14.7 mg, 144.9 pmol, 1.1 eq). The mixture was cooled to 0°C, HATU (55.1 mg, 144.9 pmol, 1.1 eq) was added and stirred at 15°C for 12 hrs. The reaction mixture was filtered and concentrated under reduced pressure to give a residue, which was purified by prep- HPLC (TFA condition) to give 7.6 mg of 231 (15.6 pmol, 11.8% yield, 99.4% purity) as a white solid.

JH NMR (400MHz, CHLOROFORM-d) d ppm 8.09 (s, 1H), 7.48 (s, 1H), 7.43 (s, 2H), 7.32 (s, 1H), 7.19 (s, 1H), 6.74 (s, 1H), 4.51 (td, / = 3.8, 7.7 Hz, 1H), 4.02 - 3.94 (m, 2H), 3.78 (t, / = 6.5 Hz, 2H), 3.60 (ddd, / = 3.1, 8.3, 11.6 Hz, 2H), 3.40 (t, / = 7.5 Hz, 2H), 2.57 (quin, / = 7.0 Hz, 2H), 2.10 - 2.01 (m, 2H), 1.86 - 1.73 (m, 2H)

LCMS (ESI+): m/z 485.1 (M+H)

Method B: (batch 2)

Experimental Procedures:

Preparation of compound 3:

To a solution of l-chloro-3-fluoro-5-nitro-benzene (100 mg, 569.6 pmol, 1 eq ) in 2 mL of dimethyl form amide was added NaH (34.2 mg, 854.5 pmol, 60% purity, 1.5 eq) at 0°C. The mixture was stirred at 0°C for 0.5 hour. Then tetrahydropyran-4-ol (69.8 mg, 683.6 pmol, 68.5 pL, 1.2 eq) was added and stirred at 20°C for 11.5 hours. After reaction, it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 150 mg of crude compound 3 as a yellow oil, which was used directly into the next step.

To a solution of 4-(3-chloro-5-nitro-phenoxy)tetrahydropyran (150 mg, 582.1 pmol, 1 eq) in 5 mL of ethyl alcohol and 1 mL of water was added Fe (162.5 mg, 2.9 mmol, 5 eq) and NH4CI (155.7 mg, 2.9 mmol, 5 eq). The mixture was stirred at 80°C for 1 hour. Then it was filtered, the filtrate was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 90 mg of crude compound 4 as a yellow oil, which was used into the next step directly. Preparation of compound 231 (batch 2):

231

To a solution of 2-chloro-5-(l,l-dioxo-l,2-thiazolidin-2-yl)benzoic acid (100 mg, 362.7 pmol, 1 eq ) in 2 mL of dimethyl form amide was added compound 5 (82.6 mg, 362.7 pmol, 1 eq ) and TEA (44.0 mg, 435.2 mihoI, 1.2 eq). The mixture was cooled to 0°C, HATU (165.5 mg, 435.2 mihoI, 1.2 eq) was added and then stirred at 25 °C for 12 hours. After reaction, the reaction mixture was filtered, the filtrate was purified twice by prep- HPLC (TFA condition) to give 25.4 mg of 231 (51.6 pmol, 14.2% yield, 98.6% purity) as a white solid.

J H NMR (400MHz, DMSO-r/6) d ppm 10.69 (br s, 1H), 7.58 (br d, 7=8.0 Hz, 1H), 7.43 (br s, 1H), 7.38 - 7.23 (m, 3H), 6.87 (br s, 1H), 4.58 (br s, 1H), 3.80 (br d, 7=19.8 Hz, 4H), 3.60 - 3.47

(m, 4H), 2.43 - 2.37 (m, 2H), 1.95 (br s, 2H), 1.59 (br s, 2H)

LCMS (ESI+): m/z 485.0 (M+H)

Method C: (batch 3, scale up reaction)

Experimental Procedures: Preparation of compound 3:

To a solution of 3-chloro-5-nitro-phenol (12 g, 69.1 mmol, 1 eq ) in 120 mL of dichloromethane was added PPI13 (21.7 g, 82.9 mmol, 1.2 eq) and tetrahydropyran-4-ol (7.8 g, 76.1 mmol, 7.6 mL, 1.1 eq). Then DIAD (16.8 g, 82.9 mmol, 16.1 mL, 1.2 eq) was added at

0°C. The mixture was stirred at 25°C for 12 hours. Then it was partitioned between 500 mL of 1M NaOH and 1000 mL of ethyl acetate. The organic phase was separated, washed with 500 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =10: 1 to 5: 1) to give 12 g of compound 3 (46.6 mmol, 67.4% yield) as a yellow oil.

Preparation of compound 4:

To a solution of 4-(3-chloro-5-nitro-phenoxy)tetrahydropyran (14 g, 54.3 mmol, 1 eq) in

80 mL of ethyl alcohol and 10 mL of water was added Fe (15.2 g, 271.7 mmol, 5 eq) and NH4CI (14.5 g, 271.7 mmol, 5 eq). The mixture was stirred at 80 °C for 12 hours. After reaction, it was filtered, the filtrate was concentrated under reduced pressure to give a residue, which was partitioned between 500 mL of water and 1L of ethyl acetate. The organic phase was separated, washed with 500 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether : ethyl acetate =10:1 to 5: 1) to give 8.8 g of compound 4 (38.6 mmol, 71.1% yield) as a yellow oil. Preparation of compound 231 (batch 3):

4

To a solution of 3-chloro-5-tetrahydropyran-4-yloxy-aniline (20 g, 87.8 mmol, 1 eq) in 500 mL of dimethyl form amide was added compound 5 (24.2 g, 87.8 mmol, 1 eq) and TEA (10.7 g, 105.4 mmol, 14.7 mL, 1.2 eq). The mixture was cooled to 0°C, HATU (40.1 g, 105.4 mmol, 1.2 eq) was added and stirred at 25°C for 5 hours. After reaction, the mixture was added dropwise into 400 mL of water with stirring. The resulting mixture was filtered and the filter cake was washed with 50 mL of methanol, and dried in vacuum to give a crude product. The crude product was dissolved in 500 mL of dichloromethane at 50°C and stirred at 80°C for 12 hours. Then the mixture was cooled to 25 °C with stirring for 0.5 hour. The mixture was filtered and the filter cake was dried in vacuum to get a off-white solid, which was added 500 mL of water and stirred at 100°C for another 12 hours. Then the mixture was filtered and the filter cake was washed for three times with 300 mL of water, dried in the vacuum oven at 80°C over 6 hours to give 22 g of crude product, which was then dissolved in 400 mL of dichloromethane and 400 mL of methanol at 50°C. The solution was stirred at 70°C for 0.5 hour and then concentrated under reduced pressure to give a residue, which was added 2 L of methanol and stirred at 80°C for 6 hours. The resulting mixture was concentrated under reduced pressure to give a residue, which was added into 2 L of water and stirred at 110°C for 10 hours. Then the mixture was filtered and dried in the vacuum oven to give 20.3 g of 231 (41.7 mmol, 91.9% yield, 99.5% purity) as a off-white solid.

J H NMR (400MHz, DMSO-de) d ppm 10.66 (s, 1H), 7.57 (d, / = 8.8 Hz, 1H), 7.43 (t, / = 1.6 Hz, 1H), 7.38 - 7.25 (m, 3H), 6.86 (t, / = 1.9 Hz, 1H), 4.57 (tt, / = 4.1, 8.6 Hz, 1H), 3.89 - 3.74 (m, 4H), 3.56 (t, / = 7.4 Hz, 2H), 3.51 - 3.42 (m, 2H), 2.42 (quin, / = 6.9 Hz, 2H), 2.01 - 1.92 (m, 2H), 1.64 - 1.52 (m, 2H)

LCMS (ESI+): m/z 485.0 (M+H) Example 61: Synthesis of compound 312:

Experimental Procedures:

Preparation of compound 2:

To a solution of butan-1 -amine (416.6 mg, 5.7 mmol, 563 pL, 5 eq) in 1 mL of dimethyl sulfoxide was added K2CO3 (314.9 mg, 2.3 mmol, 2 eq) and compound 1 (200.0 mg, 1.1 mmol,

1 eq). The mixture was stirred at 70°C for 3 hours. Then it was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 260 mg of crude compound 2 as a yellow oil.

Preparation of compound 3:

To a solution of compound 2 (200 mg, 874.6 pmol, 1 eq) in 1 mL of dichloromethane was added TEA (177.0 mg, 1.8 mmol, 244 pL, 2 eq), DMAP (10.7 mg, 87.5 pmol, 0.1 eq), tert- butoxycarbonyl tert-butyl carbonate (210.0 mg, 962.1 pmol, 221 pL, 1.1 eq). The mixture was stirred at 25 °C for 2 hours. Then it was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 280 mg of crude compound 3 as a yellow oil.

Preparation of compound 4:

A mixture of compound 3 (300 mg, 912.4 pmol, 1 eq), Fe (254.8 mg, 4.6 mmol, 5 eq), NH4CI (488.1 mg, 9.1 mmol, 10 eq) in 3 mL of ethanol and 0.6 mL of H2O was stirred at 80°C for 1 hour. Then it was concentrated under reduced pressure to get a residue. The residue was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 270 mg of crude compound 4 as a yellow oil.

Preparation of compound 6:

A mixture of compound 5 (94.4 mg, 304.2 pmol, 1.0 eq), TEA (61.6 mg, 608.5 pmol, 2 eq ), compound 4 (100 mg, 334.7 pmol, 1.1 eq) in 1.5 mL of dimethyl formamide was added HATU (127.3 mg, 335 pmol, 1.1 eq) at 0°C. After the addition, the mixture was stirred at 25°C for 12 hours. Then it was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 170 mg of crude compound 6 as a yellow oil.

Preparation of compound 312:

To a solution of compound 6 (170 mg, 287.7 pmol, 1 eq) in 1 mL of dichloromethane was added 1 mL of TLA. The mixture was stirred at 25 °C for 1 hour. Then it was concentrated under reduced pressure to give the crude product. The crude product was purified by reversed- phase MPLC (TLA condition) to give 43.4 mg of compound 312 (69.6 pmol, 24.2% yield, 97.0% purity, TLA salt) as a brown solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.47 - 10.50 (m, 1 H), 7.49 - 7.53 (m, 1 H), 7.26 - 7.31 (m, 1 H), 6.92 - 6.96 (m, 1 H), 6.92 - 6.92 (m, 1 H), 6.86 - 6.90 (m, 1 H), 6.33 - 6.37 (m, 1 H), 3.76 - 3.84 (m, 2 H), 3.60 (t, / = 7.38 Hz, 2 H), 2.97 (br t, / = 6.94 Hz, 1 H), 2.94 - 2.95 (m, 1

H), 2.41 - 2.45 (m, 2 H), 1.48 - 1.55 (m, 2 H), 1.32 - 1.42 (m, 2 H), 0.91 (t, J = 7.25 Hz, 3 H).

LCMS (ESI+): m/z 490.1 (M+H) Compound 324 was prepared analogously coupling the appropriate aniline with the acid 5, followed by the removal of the Boc group. And the corresponding aniline was also made through a SNAr reaction firstly, followed by introduction of Boc group and reduction of nitro group.

ci

Compound 324:

J H NMR (400 MHz, DMSO-de) d ppm 10.62 - 10.70 (m, 1 H), 7.49 - 7.54 (m, 1 H), 7.28 - 7.33 (m, 1 H), 7.21 - 7.24 (m, 1 H), 7.16 - 7.21 (m, 1 H), 6.64 - 6.68 (m, 1 H), 6.19 - 6.40 (m, 1 H), 3.76 - 3.85 (m, 2 H),3.55 - 3.65 (m, 2 H), 2.95 - 3.04 (m, 2 H), 2.38 - 2.46 (m, 2 H), 1.47 - 1.57 (m, 2 H), 1.31 - 1.43 (m, 2 H), 0.86 - 0.96 (m, 3 H).

LCMS (ESI+) : m/z 481.1 (M+H)

Compound 322 was prepared analogously coupling the commercial 3-aminobenzonitrile with the acid 5.

Compound 322:

JH NMR (400 MHz, DMSO-de) d ppm 10.94 - 11.08 (m, 1 H), 8.14 - 8.17 (m, 1 H), 7.89 - 7.94 (m, 1 H), 7.58 (br s, 2 H), 7.52 - 7.56 (m, 1 H), 7.35 - 7.38 (m, 1 H), 3.77 - 3.86 (m, 2 H), 3.57 - 3.64 (m, 2 H), 2.38 - 2.47 (m, 2 H).

LCMS (ESI+): m/z 410.0 (M+H)

Example 62: Synthesis of compound 325:

Experimental Procedures:

Preparation of compound 2:

To a solution of compound 1 (100 mg, 602.0 pmol, 1 eq ) in 3 mL of DMSO was added N-methylbutan-1 -amine (157.4 mg, 1.8 mmol, 213.9 pL, 3 eq) and K2CO3 (166.4 mg, 1.2 mmol, 2 eq). The mixture was stirred at 70°C for 3 hours. Then it was partitioned between 10 mL of water and 80 mL of ethyl acetate. The organic phase was separated, dried over anhydrous

Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to get 140 mg of crude compound 2 as yellow oil, which was used directly into the next step without purification.

Preparation of compound 3:

To a solution of compound 2 (140 mg, 600.2 pmol, 1 eq) in 3 mL of ethanol and 0.6 mL of water was added Fe (167.6 mg, 3.0 mmol, 5 eq) and NH4CI (321.0 mg, 6.0 mmol, 10 eq). The mixture was stirred at 80°C for 1 hour. Then it was filtered and the filtrate was partitioned between 10 mL of water and 60 mL of ethyl acetate. The organic phase was separated, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to get 120 mg of crude compound 3 as yellow oil, which was used directly into the next step without purification.

Preparation of compound 325:

3 325

To a solution of compound 3 (120 mg, 590.3 m mol, 1 eq ) in 2 mL of dimethyl form amide was added compound 4 (201.4 mg, 649.3 pmol, 1.1 eq), TEA (179.2 mg, 1.8 mmol, 3 eq) and HATU (246.9 mg, 649.3 umol, 1.1 eq) at 0°C. The mixture was stirred at 30°C for 12 hours.

The reaction mixture was filtered. The filtrate was purified by prep- HPLC (TFA condition) to get 130.5 mg of compound 325 (206.2 pmol, 34.9% yield, 96.3% purity) as light yellow soild. J H NMR (400 MHz, DMSO-de) d ppm 10.72 - 10.70 (m, 1 H), 7.55 - 7.51 (m, 1 H), 7.40 - 7.36

(m, 1 H), 7.35 - 7.30 (m, 1 H), 7.27 - 7.23 (m, 1 H), 6.88 - 6.79 (m, 1 H), 3.85 - 3.78 (m, 2 H),

3.63 - 3.56 (m, 2 H), 3.37 - 3.28 (m, 2 H), 2.94 - 2.89 (m, 3 H), 2.47 - 2.38 (m, 2 H), 1.53 - 1.43 (m, 2 H), 1.36 - 1.24 (m, 2 H), 0.94 - 0.87 (m, 3 H).

LCMS (ESI+) : m/z 495.1 (M+H)

The following compounds were prepared analogously to compound 325 using different amines:

Compound 327 J H NMR (400MHz, DMSO-de) d ppm 10.76 (s, 1 H), 7.55 (d, J = 2.8 Hz, 1 H), 7.39 (s, 1 H), 7.34 (d, J = 2.8 Hz, 1 H), 7.27 (t, / = 2.0 Hz, 1 H), 6.89 (dd, / = 1.3, 2.3 Hz, 1 H), 3.83 (t, / = 6.5 Hz, 2 H), 3.65 - 3.63 (m, 2 H), 3.42 (q, / = 7.0 Hz, 2 H), 2.91 (s, 3 H), 2.44 (t, / = 7.1 Hz, 2 H), 1.06 (t, / = 6.9 Hz, 3 H)

LCMS (ESI+) : m/z 467.1 (M+H) ci

Compound 326:

J H NMR (400 MHz, DMSO-de) d ppm 10.72 (s, 1 H), 7.54 (d, / = 2.7 Hz, 1 H), 7.39 (s, 1 H), 7.33 (d, / = 2.7 Hz, 1 H), 7.24 (s, 1 H), 6.86 (d, / = 0.9 Hz, 1 H), 3.82 (t, J = 6.5 Hz, 2 H), 3.61 (t, J = 7.3 Hz, 2 H), 3.30 (br t, / = 7.4 Hz, 2 H), 2.93 (s, 3 H), 2.44 (br t, / = 7.0 Hz, 2 H), 1.59 -

1.47 (m, 2 H), 0.88 (t, / = 7.3 Hz, 3 H).

LCMS (ESI+): m/z 481.1 (M+H)

Example 63: Synthesis of compound 315:

315 Experimental Procedures:

Preparation of compound 2:

2

To a solution of 2-allyl-4,4,5,5-tctramcthyl- 1 ,3,2-dioxaborolanc (214.9 mg, 1.3 mmol, 1.1 eq), 3-bromoaniline (200 mg, 1.2 mmol, 127 pL, 1 eq ) and K2CO3 (482.1 mg, 3.5 mmol, 3 eq ) in 1 mL of H2O and 5 mL of dioxane was added Pd(dppf)Ch (85.1 mg, 116.3 pmol, 0.1 eq) under N2. The mixture was stirred for 12 hours at 110°C. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate = 5: 1 to 3: 1) to get 118.4 mg compound 2 (888.9 pmol, 76.6% yield) as a brown oil.

Preparation of compound 3:

To a solution of 3-allylaniline (118.4 mg, 888.9 pmol, 1 eq) in 5 mL of ethyl acetate was added Pd/C (20 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with ¾ several times. The mixture was stirred under ¾ (15 psi) at 25 °C for 1 hour. Then it was filtered and the filtrate was concentrated under reduced pressure to get 110 mg of crude compound 3 as a yellow oil.

Preparation of compound 315:

3 315

To solution of compound 4 (277.6 mg, 894.9 m ihoI, 1.1 eq ) and 3-propylaniline (110 mg, 813.6 pmol, 1 eq) in 2 mL of dimethyl formamide was added TEA (164.7 mg, 1.6 mmol, 2 eq). Then the mixture was cooled to 0°C and HATU (340.3 mg, 894.9 pmol, 1.1 eq) was added in one portion. The mixture was allowed to warm to to 25 °C and stirred for 12 hours. Then it was filtered and the filtrate was concentrated under reduced pressure to give a residue, which was purified by prep- HPLC (TFA condition) to get 93.9 mg of compound 315 (219.7 pmol, 27.0% yield, 100% purity) as a pink solid.

JH NMR (400 MHz, DMSO-de) d ppm 0.90 (t, 7 =7.34 Hz, 3 H), 1.58 (sxt, 7 =7.41 Hz, 2 H), 2.42 (quin, 7 =6.94 Hz, 2 H), 2.51 - 2.57 (m, 2 H), 3.59 (t, 7 =7.40 Hz, 2 H), 3.80 (t, 7 =6.48 Hz, 2 H), 6.95 (d, 7 =7.58 Hz, 1 H), 7.22 - 7.32 (m, 2 H), 7.44 - 7.58 (m, 3 H), 10.54 (s, 1 H).

LCMS (ESI+): m/z 426.9 (M+H)

The amines of following compounds were made analogously introduction of propyl through a Suzuki reaction, followed by hydrogenation of the double bond. The obtained aniline was then coupled with acid 4:

Compound 316:

J H NMR (400 MHz, DMSO-de) d ppm 0.89 (t, 7=7.27 Hz, 3 H), 1.57 (sxt, 7=7.41 Hz, 2 H), 2.27 (s, 3 H), 2.42 (t, 7=7.03 Hz, 2 H), 2.51 (br s, 2 H), 3.59 (t, 7=7.40 Hz, 2 H), 3.81 (t, 7=6.48 Hz, 2

H), 6.78 (s, 1 H), 7.28 (d, 7=2.69 Hz, 1 H), 7.33 (s, 2 H), 7.44 - 7.54 (m, 1 H), 10.46 (s, 1 H).

LCMS (ESI+): m/z 440.9 (M+H) Compound 317:

J H NMR (400 MHz, DMSO-de) d ppm 0.89 (t, /=7.34 Hz, 3 H), 1.60 (sxt, /=7.43 Hz, 2 H), 2.37 - 2.48 (m, 1 H), 2.38 - 2.45 (m, 1 H), 2.39 - 2.47 (m, 1 H), 2.39 - 2.47 (m, 1 H), 2.39 - 2.47 (m, 1 H), 2.61 (t, J=l.52 Hz, 2 H), 3.60 (t, /=7.34 Hz, 2 H), 3.81 (t, /=6.48 Hz, 2 H), 7.35 (d, /= 2.69 Hz, 1 H), 7.47 (s, 1 H), 7.54 (d, /= 2.69 Hz, 1 H), 7.78 (s, 1 H), 7.95 (s, 1 H), 10.93 (s, 1 H).

LCMS (ESI+): m/z 452.0 (M+H)

Compound 318:

J H NMR (400 MHz, DMSO-de) d ppm 0.90 (t, /=7.34 Hz, 3 H), 1.58 (sxt, /=7.43 Hz, 2 H), 2.39 - 2.45 (m, 2 H), 2.52 (br s, 2 H), 3.60 (t, /=7.34 Hz, 2 H), 3.73 (s, 3 H), 3.81 (t, /=6.46 Hz, 2 H),

6.55 (s, 1 H) 7.11 (s, 1 H), 7.16 - 7.20 (m, 1 H), 7.29 (d, /= 2.76 Hz, 1 H), 7.51 (d, /=2.64 Hz, 1 H), 10.54 (s, 1 H).

LCMS (ESI+): m/z 457.0 (M+H)

Compound 314:

J H NMR (400 MHz, DMSO-de) d = 10.91 (s, 1H), 7.98 (s, 1H), 7.75 (s, 1H), 7.55 (d, J = 2.6 Hz, 1H), 7.39 - 7.30 (m, 2H), 3.82 (t, J = 6.5 Hz, 2H), 3.61 (t, / = 7.3 Hz, 2H), 2.65 (br t, / = 7.5 Hz, 2H), 2.47 - 2.40 (m, 2H), 1.62 (sxt, / = 7.4 Hz, 2H), 0.91 (t, / = 7.3 Hz, 3H).

LCMS (ESI+): m/z 495.0 (M+H) Compound 323:

J H NMR (400 MHz, DMSO-de) d = 10.80 (s, 1H), 8.20 (s, 1H), 7.77 (s, 1H), 7.58 - 7.51 (m, 2H), 7.34 (d, 7 = 2.4 Hz, 1H), 3.86 (s, 3H), 3.81 (br t, 7 = 6.4 Hz, 2H), 3.61 (t, 7 = 7.3 Hz, 2H), 2.63 (br t, 7 = 7.4 Hz, 2H), 2.43 (s, 2H), 1.65 - 1.56 (m, 2H), 0.90 (t, 7 = 7.3 Hz, 3H).

LCMS (ESI+): m/z 485.1 (M+H)

J H NMR (400 MHz, DMSO-de) d ppm 0.91 (t, 7=7.25 Hz, 3 H), 1.58 - 1.66 (m, 2 H), 2.44 (t, 7=6.94 Hz, 2 H), 2.66 (t, 7=7.57 Hz, 2 H), 3.62 (t, 7=7.32 Hz, 2 H), 3.82 (t, 7=6.50 Hz, 2 H), 7.29 (d, 7=8.00 Hz, 1 H), 7.35 (d, 7=2.63 Hz, 1 H), 7.52 (s, 1 H), 7.54 (d, 7=2.63 Hz, 1 H),

7.79 (d, 7=8.00 Hz, 1 H), 10.92 (s, 1 H).

LCMS (ESI+): m/z 452.1 (M+H)

Example 64: Synthesis of compound 330:

330 Experimental Procedures:

Preparation of compound 1:

323 1

To a solution of compound 323 (400 mg, 824.1 pmol, 1 eq ) in 4 mL of methanol and 0.4 mL of water was added NaOH (65.9 mg, 1.7 mmol, 2 eq). The mixture was stirred at 50°C for 12 hours. Then it was partitioned between 10 mL of 1M HC1 and 25 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 350 mg of crude compound 1 as a yellow solid.

Preparation of 330:

To a solution of compound 1 (45 mg, 95.5 umol, 1 eq) in 2 mL of dimethyl form amide was added TEA (14.5 mg, 143.2 pmol, 1.5 eq) and HATU (43.6 mg, 114.6 pmol, 1.2 eq). The mixture was stirred at 25°C for 1 hour. Meanwhile, a solution of methanamine hydrochloride (19.3 mg, 286.4 pmol, 3 eq) in 1 mL of dimethyl formamide was added TEA (48.3 mg, 477.4 pmol, 5 eq) and stirred at 25 °C for 1 hour. Then the mixture was added into above reaction mixture and stirred at 25 °C for additional 10 hours. After reaction, it was filtered to give the filtrate, which was purified by reversed-phase MPLC (TFA condition) to give 4.1 mg of compound 330 (8.2 pmol, 8.6% yield, 97.1% purity) as a white solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.71 (s, 1H), 8.40 (br d, / = 4.5 Hz, 1H), 7.95 (s, 1H), 7.67 (s, 1H), 7.52 (d, / = 2.6 Hz, 1H), 7.42 (s, 1H), 7.33 (d, J = 2.6 Hz, 1H), 3.82 (s, 2H), 3.61 (t, / = 7.3 Hz, 2H), 2.78 (d, / = 4.5 Hz, 3H), 2.60 (t, / = 7.5 Hz, 2H), 2.44 (t, / = 6.9 Hz, 2H), 1.70 - 1.55 (m, 2H), 0.92 (t, / = 7.3 Hz, 3H).

LCMS (ESI+): m/z 484.1 (M+H)

Compound 329 was prepared analogously coupling ammonium chloride with acid 1:

Compound 329:

J H NMR (400 MHz, DMSO-de) d ppm 10.68 (s, 1H), 7.98 - 7.88 (m, 2H), 7.68 (s, 1H), 7.52 (d, J = 2.6 Hz, 1H), 7.47 (s, 1H), 7.32 (d, J = 2.6 Hz, 2H), 3.82 (t, J = 6.4 Hz, 2H), 3.60 (t, J = 7.3 Hz, 2H), 2.59 (t, / = 7.6 Hz, 2H), 2.43 (br t, / = 7.0 Hz, 2H), 1.70 - 1.57 (m, 2H), 0.92 (t, / = 7.3 Hz, 3H).

Example 65: Synthesis of compound 334:

5 6 334

Experimental Procedures:

Preparation of compound 2:

To a solution of compound 1 (30 g, 123.9 mmol, 1 eq) in 500 mL of ethanol was added NaNCh (61.6 g, 892.5 mmol, 7.2 eq) and H2SO4 (92.0 g, 938.1 mmol, 50 mL, 7.6 eq). The mixture was stirred at 90°C for 12 hours. The reaction mixture was concentrated under reduced pressure, the residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate =1 :0 to 10:1) to give 25 g of compound 2 (110.1 mmol, 88.8% yield) as a yellow solid.

Preparation of compound 3:

To a solution of compound 2 (10 g, 44.1 mmol, 1 eq) in 100 mL of ethanol and 10 mL of water was added Fe (4.9 g, 88.1 mmol, 2 eq) and NH4CI (7.1 g, 132.2 mmol, 3 eq). The mixture was stirred at 80°C for 12 hours. Then it was filtered and the filtrate was concentrated under reduced pressure to give a residue, which was partitioned between 500 mL of water and 1 L of ethyl acetate. The organic phase was separated, washed with 500 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 8 g of crude compound 3 as a yellow solid.

Preparation of compound 5:

To a mixture of compound 4 (92.9 mg, 446.6 pmol, 1.1 eq), compound 3 (80 mg, 406.0 pmol, 1 eq), and K2CO3 (56.1 mg, 406.0 mihoI, 1 eq) in 0.5 mL of H2O and 3 mL of dioxane was added Pd(dppf)Cl2 (29.7 mg, 40.6 mihoI, 0.1 eq) and purged with N2 for 3 times, and then the mixture was stirred at 100°C for 12 hours under N2 atmosphere. After reaction, it was partitioned between 5 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated in vacuum. The residue was purified by prep- TLC (Si(¾, eluting with ethyl petroleum ether : ethyl acetate= 2: 1) to get 60 mg of compound 5 (302.6pmol, 74.5% yield) as a brown oil.

Preparation of compound 6:

To a solution of compound 5 (60 mg, 302.6 pmol, 1 eq) in 10 mL of ethyl acetate was added Pd/C (20 mg, 10% purity) under N2 atmosphere. The suspension was degassed and purged with ¾ for 3 times. The mixture was stirred under ¾ (15 psi) at 20°C for 1 hour. The mixture was filtered and the filtrate was concentrated in vacuum to get 60 mg of crude compound 6 as a yellow solid, which was used into the next step directly without purification.

Preparation of compound 334:

6 334

To a solution of compound 7 (102.2 mg, 329.5 pmol, 1.1 eq), compound 6 (60 mg, 299.6 pmol, 1 eq) and TEA (90.9 mg, 898.7 mihoI, 3 eq) in 3 mL of dimethyl form amide was added HATU (125.3 mg, 329.5 mihoI, 1.1 eq) at 0°C. After addition, the mixture was stirred at 20°C for 3 hours. After reaction, the mixture was filtered and concentrated in vacuum. The residue was purified by prep- HPLC (TFA condition) to get 42.6 mg of compound 334 (86.2 pmol,

28.8% yield, 99.5% purity) as a white solid.

J H NMR (400 MHz, DMSO-de) d ppm 1.18 - 1.29 (m, 1 H), 1.30 - 1.45 (m, 4 H), 1.70 (br d, / = 11.88 Hz, 1 H), 1.80 (br d, /=7.00 Hz, 4 H), 2.43 (t, / = 6.94 Hz, 2 H), 2.56 (br d, / = 11.26 Hz, 1 H), 3.60 (t, 7 = 7.32 Hz, 2 H), 3.81 (t, 7 = 6.44 Hz, 2 H), 7.35 (d, 7 = 2.63 Hz, 1 H), 7.49 (s, 1 H), 7.54 (d, 7 = 2.50 Hz, 1 H), 7.83 (s, 1 H), 7.95 (s, 1 H), 10.93 (s, 1 H).

LCMS (ESI+): m/z 492.0 (M+H)

The following compounds were prepared analogously coupling the appropriate borate with bromide 3, followed by hydrogenation and amide coupling reaction:

Compound 335:

J H NMR (400 MHz, DMSO-de) d ppm 1.59 - 1.69 (m, 2 H), 1.69 - 1.78 (m, 2 H), 2.43 (br t, 7=6.94 Hz, 2 H), 2.79 - 2.91 (m, 1 H), 3.38 - 3.49 (m, 2 H), 3.56 - 3.66 (m, 2 H), 3.81 (t, 7=6.44

Hz, 2 H), 3.95 (br dd, 7=11.07, 3.19 Hz, 2 H), 7.35 (d, 7=2.63 Hz, 1 H), 7.54 (d, 7=2.25 Hz, 2 H), 7.85 (s, 1 H), 7.98 (s, 1 H), 10.95 (s, 1 H)

LCMS (ESI+): m/z 494.1 (M+H)

Compound 328:

JH NMR (400MHz, DMSO-de) d ppm 10.95 (s, 1H), 7.96 (s, 1H), 7.80 (s, 1H), 7.54 (d, 7 = 2.6 Hz, 1H), 7.49 (s, 1H), 7.35 (d, 7 = 2.8 Hz, 1H), 3.81 (t, 7 = 6.4 Hz, 2H), 3.61 (t, 7 = 7.3 Hz, 2H), 2.66 (q, 7=7.5 Hz, 2H), 2.43 (t, 7 = 6.9 Hz, 2H), 1.19 (t, 7 = 7.6 Hz, 3H).

LCMS (ESI+): m/z 438.0 (M+H)

Example 66: Synthesis of compound 336:

A mixture of 3-amino-5-bromo-benzonitrile (70 mg, 355.3 mitioI, 1 eq), 3-pyridylboronic acid (65.5 mg, 532.9 miti I, 1.5 eq), K2CO3 (147.3 mg, 1.1 mmol, 3 eq), Pd(dppf)Cl2 (26.0 mg, 35.5 mitioI, 0.1 eq) and 0.5 mL of water in 2 mL of dioxane was degassed and purged with N2 for 3 times, and then the mixture was stirred at 100°C for 3 hours under N2 atmosphere. Then it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (SiCh, eluting with a gradient of petroleum ether:ethyl acetate =10: 1 to 0: 1) to give 55 mg of compound 2 (55 mg, 281.7 mitioI, 79.3% yield) as a white solid.

Preparation of compound 336:

2 336 To a solution of compound 2 (55 mg, 281.7 mihoI, 1 eq ) in 2 mL of dimethyl form amide was added compound 3 (87.4 mg, 281.7 pmol, 1 eq) and TEA (42.8 mg, 422.6 mihoI, 1.5 eq). The mixture was cooled to 0°C. The mixture was added HATU (160.7 mg, 422.6 mihoI, 1.5 eq) and stirred at 20°C for 3 hours. Then it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (TLA condition) to give compound 336 (63.5 mg, 101.5 pmol, 36.0% yield, 96.1% purity, TLA salt) as a white solid.

J H NMR (400MHz, DMSO-de) d ppm 11.18 (s, 1H), 9.00 (d, J=1.8 Hz, 1H), 8.73 (d, / = 3.9 Hz, 1H), 8.32 - 8.25 (m, 2H), 8.18 (s, 1H), 8.08 (s, 1H), 7.70 (dd, / = 5.1, 7.9 Hz, 1H), 7.56 (d, / = 2.6 Hz, 1H), 7.39 (d, / = 2.8 Hz, 1H), 3.82 (t, / = 6.4 Hz, 2H), 3.62 (t, / = 7.3 Hz, 2H), 2.46 - 2.41 (m, 2H), 2.46 - 2.39 (m, 1H).

LCMS (ESI+): m/z 487.1 (M+H)

Compound 337 was prepared analogously coupling the appropriate boric acid with the bromide 1, followed by an amide coupling reaction with acid 2:

Compound 337:

J H NMR (400MHz, DMSO-de) d ppm 11.23 (s, 1H), 8.86 (d, / = 6.1 Hz, 2H), 8.42 (s, 1H), 8.21 (d, / = 8.4 Hz, 2H), 8.01 (d, / = 6.1 Hz, 2H), 7.56 (d, / = 2.6 Hz, 1H), 7.40 (d, / = 2.6 Hz, 1H), 3.82 (t, / = 6.5 Hz, 2H), 3.62 (t, / = 7.3 Hz, 2H), 2.44 (t, / = 6.9 Hz, 2H).

LCMS (ESI+): m/z 487.1 (M+H)

Example 67: Synthesis of compound 338:

Experimental Procedures:

Preparation of compound 3:

To a solution of compound 1 (50 mg, 253.8 pmol, 1 eq) in 1 mL of dimethyl formamide was added compound 2 (78.7 mg, 253.8 pmol, 1 eq), HATU (106.1 mg, 279.2 pmol, 1.1 eq), TEA (51.4 mg, 507.5 pmol, 2 eq). The mixture was stirred at 30°C for 12 hours. The reaction mixture was added dropwise into 2 mL of water with stirring, filtered and the filter cake was dissolve in 2 mL of ethyl acetate, dried over Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 120 mg of crude compound 3 as a yellow solid.

Preparation of compound 338:

3 338

To a solution of compound 3 (120 mg, 245.3 pmol, 1 eq ) in 2 mL of toluene was added tributyl(2-pyridyl)stannane (90.3 mg, 245.3 pmol, 1 eq), Pd(PPh3)2Ch (17.2 mg, 24.5 pmol, 0.1 eq) and purged with N2 for 3 times. The mixture was stirred at 100°C for 12 hours. The reaction mixture was filtered and concentrated under reduced pressure to give the crude product, which was purified twice by prep- HPLC (TFA condition) to give 9.9 mg of compound 338 (19.2 pmol, 7.9% yield, 94.8% purity) as a white solid.

J H NMR (400 MHz, DMSO-de) d ppm 11.13 - 11.19 (m, 1 H), 8.68 - 8.79 (m, 2 H), 8.27 - 8.34 (m, 1 H), 8.16 - 8.24 (m, 1 H), 8.04 - 8.12 (m, 1 H), 7.93 - 8.01 (m, 1 H), 7.53 - 7.57 (m, 1 H), 7.43 - 7.48 ( m , 1 H), 7.38 - 7.42 (m, 1 H), 3.78 - 3.86 (m, 2 H), 3.59 - 3.65 (m, 2 H), 2.45 (s, 2

H).

LCMS (ESI+): m/z 487.0 (M+H)

Example 68: Synthesis of compound 333:

Experimental Procedures:

Preparation of compound 3:

1

To a solution of compound 1 (50 mg, 253.8 pmol, 1 eq ) and compound 2 (45.2 mg, 279.1 pmol, 1.1 eq) in 0.1 mL of ¾0 and 1 mL of toluene was added Pd(dppf)Cl2 (18.6 mg, 25.4 pmol, 0.1 eq) and t-BuOK (85.4 mg, 761.3 pmol, 3 eq) and purged with N2 for 3 times. The mixture was stirred for 100°C for 12 hours under N2. After reaction, it was concentrated under reduced pressure to give the crude product, which was purified by prep-TLC (S1O2, eluting with petroleum ether : ethyl acetate= 3: 1) to get 40 mg of crude compound 3 as colourless oil. Preparation of compound 333:

3 333

To a solution of compound 3 (40 mg, 232.3 pmol, 1 eq ) in 1 mL of dimethyl formamide was added compound 4 (79.2 mg, 255.5 pmol, 1.1 eq), TEA (70.5 mg, 696.8 pmol, 3 eq), and HATU (97.1 mg, 255.5 pmol, 1.1 eq) at 0°C. The mixture was stirred at 35°C for 12 hours. After reaction, the reaction mixture was filtered. The filtrate was purified by prep- HPLC (TFA condition) to get 9.7 mg of compound 333 (20.9 pmol, 9.0% yield, 100.0% purity) as light yellow soild.

JH NMR (400 MHz, DMSO-de) d = 10.98 - 10.91 (m, 1 H), 7.98 - 7.95 (m, 1 H), 7.82 - 7.76 (m, 1 H), 7.56 - 7.52 (m, 1 H), 7.49 - 7.43 (m, 1 H), 7.37 - 7.32 (m, 1 H), 3.85 - 3.78 (m, 2 H), 3.64 - 3.59 (m, 2 H), 3.58 - 3.52 (m, 1 H), 2.47 - 2.38 (m, 2 H), 2.36 - 2.26 (m, 2 H), 2.15 - 1.93 (m, 3 H), 1.88 - 1.75 (m, 1 H).

LCMS (ESI+): m/z 464.1 (M+H)

Compound 332 was prepared analogously coupling appropriate borate with bromide 1, followed by an amide coupling reaction with acid 4:

Compound 332:

1H NMR (400 MHz, DMSO-de) d ppm 10.84 - 10.93 (m, 1 H), 7.84 - 7.98 (m, 1 H), 7.62 - 7.67 (m, 1 H), 7.47 - 7.57 (m, 1 H), 7.26 - 7.37 (m, 2 H), 3.75 - 3.86 (m, 2 H), 3.56 - 3.63 (m, 2 H),

2.41 - 2.46 (m, 2 H), 1.96 - 2.04 (m, 1 H), 0.99 - 1.06 (m, 2 H), 0.71 - 0.77 (m, 2 H).

LCMS (ESI+): m/z 450.0 (M+H) Example 69: Synthesis of compound 311:

Experimental Procedures:

Preparation of compound 2:

To a solution of N-methylbutan-1 -amine (248.3 mg, 2.9 mmol, 337.3 pL, 5 eq) in 1 mL of dimethyl sulfoxide was added K2CO3 (157.5 mg, 1.1 mmol, 2 eq) and compound 1 (100 mg, 569.7 pmol, 1 eq). The mixture was stirred at 70°C for 3 hours. Then it was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 130 mg of crude compound 2 as a yellow oil. Preparation of compound 3:

To a solution of compound 2 (130 mg, 535.6 pmol, 1 eq) in 1 mL of ethanol and 0.2 mL of water was added Fe (149.6 mg, 2.7 mmol, 5 eq) and NH4CI (286.5 mg, 5.4 mmol, 10 eq). The mixture was heated to 80°C and stirred for 1 hour. Then it was filtered and the filtrate was concentrated under reduced pressure to get a residue, which was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 110 mg of compound 3 as a yellow oil.

Preparation of Compound 311:

3

To a solution of compound 3 (110 mg, 517.1 m mol, 1.1 eq) and compound 4 (145.8 mg, 470.1 pmol, 1 eq) in 2 mL of DMF was added TEA (142.7 mg, 1.4 mmol, 196.3 pL, 3 eq) and HATU (214.5 mg, 564.1 pmol, 1.2 eq) at 0°C. The mixture was stirred at 20°C for 12 hours.

After reaction, the mixture was filtered and the filtrate was purified by prep- HPLC (TFA condition) to give 109.7 mg of compound 311 (173.2 pmol, 36.8% yield, 97.7% purity, TFA salt) as a white solid.

J H NMR (400 MHz, DMSO-de) d ppm 10.49 - 10.60 (m, 1 H), 7.49 - 7.54 (m, 1 H), 7.27 - 7.31 (m, 1 H), 7.10 - 7.13 (m, 1 H), 6.90 - 6.94 (m, 1 H), 6.42 - 6.46 (m, 1 H), 3.77 - 3.84 (m, 2 H),

3.57 - 3.62 (m, 2 H), 3.26 (br s, 2 H), 2.84 - 2.90 (m, 3 H), 2.39 - 2.45 (m, 2 H), 1.43 - 1.52 (m, 2 H), 1.24 - 1.34 (m, 2 H), 0.87 - 0.95 (m, 3 H). LCMS (ESI+): m/z 504.1 (M+H)

The following compounds were prepared analogously by using different amines to do SNAr reaction firstly, followed by reduction of the nitro group and amide coupling reaction:

Compound 310:

J H NMR (400 MHz, DMSO-de) d ppm 10.55 (s, 1 H), 7.49 - 7.54 (m, 1 H), 7.27 - 7.32 (m, 1 H), 7.10 - 7.15 (m, 1 H), 6.88 - 6.94 (m, 1 H), 6.42 - 6.48 (m, 1 H), 3.81 (br t, / = 6.25 Hz, 2 H), 3.60 (br t, / = 7.19 Hz, 2 H), 3.21 - 3.28 (m, 2 H), 2.85 - 2.92 (m, 3 H), 2.39 - 2.45 (m, 2 H), 1.46 - 1.58 (m, 2 H), 0.82 - 0.92 (m, 3 H).

LCMS (ESI+) : m/z 490.1 (M+H)

Compound 309:

J H NMR (400 MHz, DMSO-de) d ppm 10.52 - 10.57 (m, 1 H), 7.49 - 7.54 (m, 1 H), 7.27 - 7.32 (m, 1 H), 7.09 - 7.14 (m, 1 H), 6.92 - 6.97 (m, 1 H), 6.45 - 6.50 (m, 1 H), 3.78 - 3.84 (m, 2 H), 3.58 - 3.62 (m, 2 H), 3.40 - 3.41 (m, 2 H), 2.83 - 2.88 (m, 3 H), 2.38 - 2.46 (m, 2 H), 1.00 - 1.08

(m, 3 H).

LCMS (ESI+): m/z 476.0 (M+H)

Example 70: Synthesis of compound 319:

Experimental Procedures:

Preparation of compound 2:

1 2

To a solution of compound 1 (945 mg, 3.3 mmol, 1 eq ) in 40 mL of t-BuOH was added dropwise DPPA (1.4 g, 4.9 mmol, 1.1 mL, 1.5 eq) and TEA (503.3 mg, 4.9 mmol, 1.5 eq) at 25°C. After addition, the mixture was stirred at this temperature for 2 hours, and then the mixture was warmed to 85°C and stirred for another 12 hours. After reaction, the mixture was filtered and concentrated in vacuum to get a residue. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate = 5: 1 to 3: 1) to get 824 mg of compound 2 (2.3 mmol, 69.8% yield) as a white gum. Preparation of compound 3:

A mixture of 2-ally 1-4, 4, 5,5-tctramcthyl- 1 ,3,2-dioxaborolanc (427.7 mg, 2.6 mmol, 1.1 eq), compound 2 (824 mg, 2.3 mmol, 1 eq), Pd(dppf)Cl2 (169.3 mg, 231.4 pmol, 0.1 eq), K2CO3 (959.3 mg, 6.9 mmol, 3 eq) in 1 mL of water and 5 mL of dioxane was degassed and purged with N2 for 3 times, and then the mixture was stirred at 100°C for 12 hours under N2 atmosphere. After reaction, the mixture was filtered and concentrated to give a residue, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate = 1 :0 to 20: 1) to get 602 mg of compound 3 (1.9 mmol, 82.0% yield) as a white gum.

Preparation of compound 4:

To a solution of compound 3 (602 mg, 1.9 mmol, 1 eq) in 20 mL of methanol was added Pd/C (25 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with ¾ several times. The mixture was stirred under ¾ (15psi) at 25 °C for 1 hour. After reaction, the mixture was filtered and concentrated under reduced pressure to give 427.4 mg of crude compound 4 as a white solid.

Preparation of compound 5:

To a solution of 5 mL of TFA and 10 mL of dichloromethane was added compound 4 (374.7 mg, 1.2 mmol, 1 eq). The mixture was stirred at 25°C for 1 hour. Then the mixture was concentrated in vacuum to get 390 mg of crude compound 5 (TFA salt) as a red oil.

Preparation of compound 319:

To a solution of compound 5 (195 mg, 889.6 pmol, 1 eq ) and compound 6 (275.9 mg, 889.6 pmol, 1 eq) in 3 mL of dimethyl formamide was added TEA (180.0 mg, 1.8 mmol, 2 eq). The mixture was cooled to 0°C and HATU (372.1 mg, 978.6 pmol, 1.1 eq) was added. The mixture was allowed to warm to 25 °C and stirred for another 12 hours. After reaction, the mixture was filtered and the filtrate was concentrated in vacuum to get a residue, which was purified by prep- HPLC (TFA condition) to get 57.4 mg of 319(112.3 pmol, 12.6% yield, 100% purity) as a white solid.

J H NMR (400 MHz, DMSO-de) d ppm 0.90 (t, J = 7.32 Hz, 3 H), 1.60 (sxt, / = 7.40 Hz, 2 H), 2.44 (quin, J = 6.91 Hz, 2 H), 2.60 (t, / = 7.50 Hz, 2 H), 3.61 (t, J = 7.32 Hz, 2 H), 3.82 (t, / = 6.50 Hz, 2 H), 6.96 (s, 1 H), 7.36 (d, / = 2.63 Hz, 1 H), 7.50 (s, 1 H), 7.55 (d, / = 2.75 Hz, 1 H), 7.67 (s, 1 H), 10.86 (s, 1 H).

LCMS (ESI+): m/z 511.0 (M+H)

Example 71: Synthesis of compound 321:

321

Experimental Procedures:

Preparation of compound 2:

To a solution of l,3-dibromo-5-chloro-benzene (1 g, 3.7 mmol, 1 eq) and tributyl(l- ethoxyvinyl)stannane (1.5 g, 4.1 mmol, 1.4 mL, 1.1 eq) in 50 mL of toluene was added Pd(PPh3)2Ch (259.6 mg, 369.9 pmol, 0.1 eq) under N2. The mixture was heated to 100°C and stirred for 12 hours. After reaction, a solution of 5 g of KF in 25 mL of H2O was added to the mixture. The mixture was stirred at 20°C for other 2 hours. Then it was filtered and the filter cake was washed with 10 mL of dichloromethane. The filtrate was concentrated under reduced pressure to give 960 mg of crude compound 2 as a yellow oil, which was used to next step without further purification.

Preparation of compound 3:

2 3

To a solution of compound 2 (4.8 g, 18.4 mmol, 1 eq ) in 30 mL of tetrahydrofuran was added 30 mL of 2M HC1. The mixture was stirred at 25°C for 2 hours. Then it was basified to pH~8 by addition of aqueous solution of sodium carbonate. The mixture was partitioned between 50 mL of water and 100 mL of dichloromethane. The organic phase was separated, washed with 25 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate =1:0 to 0:1) to get 3.2 g of compound 3 (13.7 mmol, 74.7% yield) as a light yellow oil.

Preparation of compound 4:

A mixture of compound 3 (3.2 g, 13.7 mmol, 1 eq), TEA (13.9 g, 137.1 mmol, 19.1 mL, 10 eq) and Pd(dppf)Ch (1.0 g, 1.4 mmol, 0.1 eq) in 100 mL of methanol was degassed and purged with CO (50psi) for three times, and then the mixture was stirred at 65°C for 24 hours under CO atmosphere. The reaction mixture was concentrated under reduced pressure to remove methanol. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate =1:0 to 4:1) to get 2.4 g of compound 4 (11.3 mmol, 82.4% yield) as a white solid. Preparation of compound 5

4 5

To a solution of methyl compound 4 (300 mg, 1.4 mmol, 1 eq ) in 4 mL of methanol was added NaBPL (58.7 mg, 1.6 mmol, 1.1 eq) at 0°C. The mixture was stirred at 25°C for 2 hours. After reaction, the mixture was concentrated under reduced pressure and the residue was diluted with 5 mL of saturated solution of ammonium chloride and partitioned between 5 mL of water and 10 mL of ethyl acetate. The organic layer was separated and washed with 10 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 280 mg of crude compound 5 as a yellow oil, which was used to next step without purification.

Preparation of compound 6:

5 6

To a solution of compound 5 (280 mg, 1.3 mmol, 1 eq) in 3mL of dichloromethane was added BAST (432.9 mg, 2.0 mmol, 429 pL, 1.5 eq). The mixture was stirred at 25°C for 1 hour. The reaction mixture was diluted with 10 mL of saturated solution of ammonium chloride and partitioned between 5 mL of water and 10 mL of ethyl acetate. The organic layer was separated and washed with 10 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give a residue, which was purified by column chromatography (SiCh, eluting with a gradient of petroleum ether: ethyl acetate =1:0 to 5:1), to give 160 mg of compound 6 (738.6 pmol, 56.6% yield) as a yellow oil. Preparation of compound 7:

To a solution of compound 6 (160 mg, 738.6 m mol, 1 eq ) in 2 mL of methanol and 1 mL of H2O was added NaOH (59.1 mg, 1.5 mmol, 2 eq). The mixture was stirred at 25 °C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove methanol. The residue was acidified by IN HC1 to pH~2, and then the mixture was partitioned between 10 mL of water and 20 mL of ethyl acetate. The organic layer was separated and washed with 5 mL of brine, dried over anhydrous Na2S04, filtered and the filtrate was concentrated under reduced pressure to give 100 mg of crude compound 7 as a white solid, which was used to the next step without purification.

Preparation of compound 8:

To a solution of compound 7 (100 mg, 493.6 pmol, 1 eq) in 2 mL of t-BuOH was added DPPA (203.7 mg, 740.3 pmol, 160.4 pL, 1.5 eq) and TEA (74.9 mg, 740.3 pmol, 1.5 eq) at 20°C. After addition, the mixture was stirred at this temperature for 2 hours. Then the mixture was warmed to 85°C and stirred at this temperature for 12 hours. After reaction, it was filtered and concentrated under reduced pressure to remove the solvent. The residue was partitioned between 50 mL of water and 100 mL of ethyl acetate. The organic layer was collected and washed with 100 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give a residue, which was purified by prep-TLC (SiCh, eluting with petroleum ether: ethyl acetate = 3: 1) to give 90 mg of compound 8 (328.8 pmol, 66.6% yield) as a yellow oil. Preparation of compound 9:

8 9

To a solution of compound 9 (90 mg, 328.8 pmol, 1 eq) in 2 mL of dichloromethane was added TFA (770.0 mg, 6.8 mmol, 500 pL, 20.5 eq). The mixture was stirred at 25°C for 1 hour. Then the reaction mixture was concentrated under reduced pressure to remove dichloromethane to give a residue, which was diluted with 35 mL of saturated NaHCCb solution and partitioned between 10 mL of water and 50 mL of ethyl acetate. The organic layer was separated and washed with 50 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 50 mg of crude compound 9 as a yellow oil, which was used into the next step without purification.

Preparation of compound 321:

To a solution of 3-chloro-5-(l-fluoroethyl)aniline (50 mg, 288.0 pmol, 1 eq) and compound 10 (89.3 mg, 288.0 pmol, 1 eq) and TEA (87.4 mg, 864.0 pmol, 3 eq) in 2 mL of dimethyl form amide was added HATU (131.4 mg, 345.6 pmol, 1.2 eq) at 0°C. The mixture was stirred at 25 °C for 12 hours. After reaction, the mixture was filtered and the filtrate was purified by prep- HPLC (TFA condition) to give 23.9 mg of compound 321 (51.3pmol, 17.8% yield 100.0% purity) as a white solid.

JH NMR (400 MHz, DMSO-de) d ppm 10.90 (s, 1H), 7.81 (s, 1H), 7.65 (s, 1H), 7.54 (d, 7 = 2.6

Hz, 1H), 7.34 (d, 7 = 2.6 Hz, 1H), 7.23 (s, 1H), 5.79 (q, 7 = 6.5 Hz, 1H), 5.67 (q, 7 = 6.3 Hz,

1H), 3.81 (t, 7 = 6.5 Hz, 2H), 3.60 (t, 7 = 7.3 Hz, 2H), 2.43 (t, 7 = 7.0 Hz, 2H), 1.63 - 1.59 (m, 2H), 1.54 (d, 7 = 6.4 Hz, 2H).

LCMS (ESI+): m/z 467.0 (M+H) Example 72: Synthesis of compound 339:

339 Experimental Procedures:

Preparation of compound 2:

To a solution of methyl 3-amino-5-cyano-benzoate (600 mg, 3.4 mmol, 1 eq) in 10 mL of dichloromethane was added TEA (516.9 mg, 5.1 mmol, 1.5 eq) and B0C2O (891.9 mg, 4.1 mmol, 1.2 eq), DMAP (41.6 mg, 340.6 pmol, 0.1 eq). The mixture was stirred at 25°C for 12 hours.

Then it was partitioned between 5 mL of saturated NH4CI and 15 mL of ethyl acetate. The organic phase was separated, washed with 5mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ethenethyl acetate =100:1 to 15: 1) to give 500 mg of compound 2 (500 mg, 1.8 mmol, 53.1% yield) as a yellow solid. Preparation of compound 3

To a solution of compound 2 (500.0 mg, 1.8 mmol, 1 eq ) in 2 mL of ethanol was added NaBPL (136.9 mg, 3.6 mmol, 2 eq). The mixture was stirred at 70°C for 12 hours. Then it was partitioned between 10 mL of water and 30 mL of ethyl acetate. The organic phase was separated, washed with 25 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 370 mg of compound 3 as a yellow oil, which was used to next step directly.

Preparation of compound 4:

To a solution of compound 3 (37 mg, 1.5 mmol, 1 eq) in 5 mL of dichloromethane was added DMP (948.1 mg, 2.2 mmol, 1.5 eq). The mixture was stirred at 25°C for 12 hours. Then it was partitioned between 30 mL of saturated Na2SC>3 and 50 mL of ethyl acetate. The organic phase was separated, washed with 15 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 350 mg of crude compound 4 as a yellow oil, which was used to next step directly.

Preparation of compound 5:

4 5

To a solution of compound 4 (320 mg, 1.3 mmol, 1 eq) in 5 mL of dichloromethane was added BAST (574.9 mg, 2.6 mmol, 2 eq). The mixture was stirred at 20°C for 2 hours. Then it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- TLC(SiC>2, eluting with petroleum ether : ethyl acetate =5: 1) to give 100 mg of compound 5 (372.7 pmol, 28.7% yield) as a yellow solid.

Preparation of compound 6:

To a solution of compound 5 (100 mg, 372.8 pmol, 1 eq) in 2 mL of dichloromethane was added 0.5 mL of TFA. The mixture was stirred at 20°C for 1 hour. Then it was partitioned between 3 mL of saturated NaHCCL and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give 60 mg of crude compound 6 as a yellow oil, which was used to next step directly.

Preparation of compound 339:

To a solution of compound 7 (60 mg, 193.45 pmol, 1 eq) in 2 mL of dimethyl formamide was added 3-amino-5-(difluoromethyl)benzonitrile (39.0 mg, 232.1 pmol, 1.2 eq) and TEA (23.5 mg, 232.1 pmol, 1.2 eq). The mixture was cooled to 0°C, HATU (88.3 mg, 232.1 pmol, 1.2 eq) was added and stirred at 20°C for 3 hours. After reaction, it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (TFA condition) to give 7.2 mg of compound 339 (15.2 pmol, 7.9% yield, 97.5% purity) as a white solid. J H NMR (400MHz, DMSO-de) d ppm 11.25 (s, 1H), 8.23 (br d, / = 4.8 Hz, 2H), 7.87 (s, 1H), 7.57 (d, J = 2.6 Hz, 1H), 7.41 (d, J = 2.6 Hz, 1H), 7.32 - 6.98 (m, 1H), 3.83 (t, J = 6.4 Hz, 2H), 3.63 (t, J = 7.3 Hz, 2H), 2.45 (br t, / = 6.9 Hz, 2H).

LCMS (ESI+): m/z 460.1 (M+H)

Example 73: Synthesis of compound 313:

Experimental Procedures:

Preparation of compound C:

To a solution of compound A (1 g, 4.8 mmol, 1 eq ), compound B (1.2 g, 7.3 mmol, 1.5 eq ) and K2CO3 (2.0 g, 14.5 mmol, 3 eq) in 5 mL of dioxane and 1 mL of H2O was added Pd(dppf)Cl2 (354.4 mg, 484.3 pmol, 0.1 eq) under N2. The mixture was stirred at 100°C for 12 hours. Then it was partitioned between 30 mL of water and 60 mL of ethyl acetate. The organic phase was separated, washed with 30 mL of brine, dried over Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate = 1 :0 to 1 : 1) to give 440 mg of compound C (2.6 mmol, 54.2% yield) as a brown oil.

Preparation of compound D:

To a solution of compound C (430 mg, 2.6 mmol, 1 eq ) in 50 mL of ethyl acetate was added Pd/C (400 mg, 10% purity) under N2. The suspension was degassed under vacuum and purged with ¾ several times. The mixture was stirred under ¾ (15 psi) at 25°C for 20 minutes. Then it was filtered and the filtrate was concentrated under reduced pressure to give 370 mg of crude compound D as a brown oil.

Preparation of compound 2:

To a solution of compound 1 (1 g, 5.0 mmol, 1 eq) in 20 mL of methanol was added SOCb (1.2 g, 9.9 mmol, 720 pL, 2 eq) at 25°C. The mixture was stirred at 70°C for 12 hours. After reaction, it was concentrated under reduced pressure to give a residue, which was diluted with 50 mL of sa/.NaHCCL and extracted with 90 mL of ethyl acetate. The organic phase was separated, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 1.1 g of crude compound 2 as a brown solid.

Preparation of compound 3:

To a solution of compound 2 (1.1 g, 5.1 mmol, 1 eq) and NH4CI (818.8 mg, 15.3 mmol, 3 eq) in 20 mL of ethanol and 4 mL of H2O was added Le (854.8 mg, 15.3 mmol, 3 eq). The mixture was stirred at 80°C for 12 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. Then it was partitioned between 30 mL of water and 60 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 950 mg of crude compound 3 as a yellow oil.

Preparation of compound 5:

To a solution of compound 3 (950 mg, 5.1 mmol, 1 eq ) and compound 4 (996.8 mg, 5.6 mmol, 683 pL, 1.1 eq) in 20 mL of dichloromethane was added dropwise TEA (1.6 g, 15.4 mmol, 2.1 mL, 3 eq) at 0°C. The mixture was stirred at 25 °C for 12 hours. Then the mixture was concentrated under reduced pressure to give a residue, which was diluted with 20 mL of methanol, and TEA (1.6 g, 15.4 mmol, 2.1 mL, 3 eq) added. The mixture was heated to 70°C and stirred for 3 hours. After reaction, the mixture was concentrated under reduced pressure to give a residue, which was purified by column chromatography (Si(¾, eluting with a gradient of petroleum ether: ethyl acetate = 1 :0 to 1: 1) to give 840 mg of compound 5 (2.9 mmol, 56.8% yield) as a yellow solid.

Preparation of compound 6:

To a solution of compound 5 (840 mg, 2.9 mmol, 1 eq) in 10 mL of methanol and 2 mL of H2O was added NaOH (347.9 mg, 8.7 mmol, 3 eq). The mixture was stirred at 25°C for 1 hour. Then it was concentrated under reduced pressure. The residue was diluted with 20 mL of water and acidified to pH 3 with IN HC1. Then it was filtered and filter cake was dried in vacuum to give 680 mg of compound 6 as a yellow solid. Preparation of compound 313:

6 313

To a solution of compound D (67.7 mg, 399.0 mihoI, 1.1 eq ) and compound 6 (100 mg, 362.7 pmol, 1 eq) in 2 mL of dimethyl form amide was added TEA (91.8 mg, 906.8 mihoI, 126 pL, 2.5 eq) and HATU (165.5 mg, 435.2 mihoI, 1.2 eq) at 0°C. The mixture was stirred at 20°C for 12 hours. Then it was filtered and the filtrate was purified by reversed-phase MPLC (TFA condition) to give 79.6 mg of compound 313 (182.5 pmol, 50.3% yield, 98.0% purity) as a white solid.

J H NMR (400 MHz, DMSO-cfc) d ppm 10.41 - 10.45 (m, 1 H), 7.72 - 7.79 (m, 2 H), 7.63 - 7.66 (m, 1 H), 7.52 - 7.55 (m, 1 H), 7.42 - 7.45 (m, 1 H), 7.02 - 7.07 (m, 1 H), 3.81 - 3.89 (m, 2 H),

3.55 - 3.63 (m, 2 H), 2.55 - 2.58 (m, 2 H), 2.43 - 2.47 (m, 2 H), 1.54 - 1.65 (m, 2 H), 0.86 - 0.94 (m, 3 H).

LCMS (ESI+): m/z 427.1 (M+H)

Example 74: Synthesis of compound 320:

Experimental Procedures:

Preparation of compound 2:

To a solution of 3,4-dichlorobenzoic acid (1 g, 5.2 mmol, 1 eq) in 10 mL of methanol was added SOCh (1.3 g, 10.5 mmol, 760 pL, 2 eq) at 0°C. The mixture was stirred at 70°C for 12 hours. Then it was concentrated under reduced pressure to give a residue. The residue was partitioned between 10 mL of saturated NaHCCb solution and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to get 1 g of crude compound 2 as a yellow oil. Preparation of compound 3:

To a solution of methyl 3,4-dichlorobenzoate (450 mg, 2.2 mmol, 1 eq) in 4.5 mL of H2SO4 was added HNO3 (223.7 mg, 2.4 mmol, 160 pL, 68% purity, 1.1 eq) at 0°C. The mixture was stirred at 0°C for 1 hour. Then it was added into 20 mL of ice water and stirred at 0°C for 0.2 hour. The mixture was partitioned between 20 mL of water and 20 mL of dichloromethane. The organic phase was separated, washed with 20 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to get 220 mg of crude compound 3 as a yellow oil.

Preparation of compound 4:

To a solution of compound 3 (880.0 mg, 3.52 mmol, 0.8 eq) in 0.2 mL of H2O and 1 mL of ethanol was added Fe (1.23 g, 22.0 mmol, 5 eq) and NH4CI (2.4 g, 43.9 mmol, 10 eq). The mixture was stirred at 80°C for 1 hour. Then it was concentrated under reduced pressure, the residue was partitioned between 25 mL of water and 50 mL of ethyl acetate. The organic phase was separated, washed with 25 mL of brine, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate =1 :0 to 4: 1) to get 70 mg of crude compound 4 as a white solid.

Preparation of compound 5:

To a solution of compound 4 (70 mg, 318.1 pmol, 1 eq) in 1.5 mL of dichloromethane was added 3-chloropropane-l-sulfonyl chloride (84.5 mg, 477.2 pmol, 1.5 eq) and TEA (48.3 mg, 477.2 pmol, 1.5 eq) at 0°C. The mixture was stirred at 25°C for 12 hours. After reaction, the mixture was concentrated under reduced pressure to give a residue. Then the residue was dissolved in 1.5 mL of methanol, and TEA (48.3 mg, 477.2 pmol, 1.5 eq) was added. The mixture was stirred at 70°C for 6 hours. Then it was concentrated under reduced pressure, the residue was partitioned between 10 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to get 100 mg of crude compound 5 as a yellow solid.

Preparation of compound 6:

To a solution of compound 5 (100 mg, 308.5 pmol, 1 eq) in 0.2 mL of H2O and 1 mL of methanol was added NaOH (49.4 mg, 1.2 mmol, 4 eq). The mixture was stirred at 25°C for 1 hour. Then it was concentrated under reduced pressure, the residue was partitioned between 10 mL of water and 15 mL of ethyl acetate. The aqueous phase was acidified to pH~3 with IN HC1 and extracted with 15 mL of ethyl acetate. The organic phase was separated, washed with 10 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to get 90 mg of crude compound 5 as a yellow solid.

Preparation of compound 320:

To a solution of 3-chloro-5-propyl-aniline (54.1 mg, 319.2 pmol, 1.1 eq) and compound 6 (90 mg, 290.2 pmol, 1 eq) in 1 mL of dimethyl form amide was added TEA (73.4 mg, 725.4 m mol, 2.5 eq) and HATU (132.4 mg, 348.2 m ihoI, 1.2 eq) at 0°C. The mixture was stirred at 20°C for 12 hours. Then it was partitioned between 5 mL of water and 25 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (TFA condition) to get 17.4 mg of compound 320 (37.55 pmol, 17.3% yield, 99.7 % purity) as a brown oil.

J H NMR (400 MHz, DMSO-de) d ppm 0.90 (t, J = 7.34 Hz, 3 H), 1.60 (sxt, / = 7.35 Hz, 2 H), 2.56 (br d, / = 7.65 Hz, 2 H), 3.37 - 3.43 (m, 2 H), 3.47 (t, J = 7.47 Hz, 2 H), 3.79 (t, J = 6.78 Hz, 2 H), 7.06 (s, 1 H), 7.51 (s, 1 H), 7.76 (s, 1 H), 8.08 (d, / = 2.01 Hz, 1 H), 8.27 (d, / = 2.01 Hz, 1 H), 10.52 (s, 1 H).

LCMS (ESI+): m/z 461.0 (M+H)

Compound 340 was prepared analogously coupling the appropriate amine with the acid

6:

Compound 340:

1H NMR (400 MHz, METHANOL-cL) d ppm 8.16 - 8.19 (m, 1 H) 8.09 - 8.11 (m, 1 H) 7.99 - 8.03 (m, 1 H) 7.79 - 7.82 (m, 1 H) 7.33 - 7.38 (m, 1 H) 3.80 - 3.90 (m, 2 H) 3.43 (t, J = 7.50 Hz, 2 H) 2.56 - 2.70 (m, 4 H) 1.62 - 1.75 (m, 2 H) 0.93 - 1.01 (m, 3 H)

LCMS (ESI+): m/z 452.0 (M+H)

Example 75: Synthesis of compound 341:

Experimental Procedures:

Preparation of compound 2:

To a solution of compound 1 (2 g, 10.8 mmol, 1 eq ) in 10 mL of H2SO4 (98%) was added HNO3 (1.1 g, 11.9 mmol, 788.8 pL, 68.0% purity, 1.1 eq) at 0°C. The mixture was stirred at 0°C for 1 hour. The reaction mixture was added drop wise to the mixture of 100 mL of ice- water and 200 mL of ethyl acetate, and the organic phase was separated, washed with 300 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 2.4 g crude compound 2 as a yellow oil.

Preparation of compound 3:

To a solution of compound 2 (2.4 g, 10.5 mmol, 1 eq) in 20 mL of ethanol and 4 mL of H2O was added Fe (2.9 g, 52.3 mmol, 5 eq) and NH4CI (5.6 g, 104.5 mmol, 10 eq). The mixture was stirred at 80°C for 1 hour. The reaction mixture was filtered and the filtrate was diluted with water and extracted twice with 250 mL of ethyl acetate. The combined organic phase was concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (eluting with ethyl acetate: petroleum ether = 0% - 18%) to give 1.4 g of crude compound 3 as a yellow oil.

Preparation of compound 4:

To a solution of compound 3 (200 mg, 1.0 mmol, 1 eq ) and TEA (304.1 mg, 3.0 mmol, 418.3 pL, 3 eq) in 4 mL of dichloromethane was added dropwise 3-chloropropane-l-sulfonyl chloride (266.1 mg, 1.5 mmol, 1.5 eq) at 0°C. The mixture was stirred at 30°C for 12 hours. After reaction, it was concentrated under reduced pressure to give a residue. The residue was diluted with 4 mL of methanol and TEA (304.1 mg, 3.0 mmol, 418.3 pL, 3 eq) was added. The mixture was stirred at 70°C for 12 hours. Then the reaction mixture was concentrated under reduced pressure to give a residue, which was partitioned between 10 mL of water and 75 mL of ethyl acetate. The organic phase was separated, dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under reduced pressure to give 300 mg of crude compound 4 as a yellow oil.

Preparation of compound 5:

4 5

To a solution of compound 4 (300 mg, 987.6 pmol, 1 eq) in 3 mL of ethanol and 0.6 mL of H2O was added NaOH (118.5 mg, 3.0 mmol, 3 eq). The mixture was stirred at 20°C for 2 hours. After reaction, it was concentrated under reduced pressure to give a residue. The residue was dissolved in 3 mL of water. Then the solution was acidified to pH 2 with IN HC1. Then it was filtered and the filter cake was dried to give 280 mg of crude compound 5 as a yellow solid. Preparation of compound 341:

5 341

To a solution of compound 6 (27.7 mg, 172.6 pmol, 1 eq ) in 1 mL of dimethyl form amide was added compound 5 (50 mg, 1 72.6 pmol, 1 eq), HATU (72.2 mg, 189.8 pmol, 1.1 eq), TEA (34.9 mg, 345.1 pmol, 48.0 pL, 2 eq). The mixture was stirred at 30°C for 12 hours. After reaction, the mixture was filtered, the filtrate was purified by prep- HPLC (TFA condition) to give 2.8 mg of Compound 341 (6.5 pmol, 3.8% yield, 100.0% purity) as a white solid.

J H NMR (400 MHz, METHANOL-c/4) d ppm 7.94 - 8.00 (m, 1 H), 7.71 - 7.79 (m, 1 H), 7.31 - 7.39 (m, 2 H), 7.22 - 7.29 (m, 1 H), 3.77 - 3.85 (m, 2 H), 3.41 - 3.51 (m, 2 H), 2.61 - 2.69 (m, 2

H), 2.48 - 2.57 (m, 2 H), 2.43 - 2.46 (m, 3 H), 1.63 - 1.73 (m, 2 H), 0.94 - 1.00 (m, 3 H).

LCMS (ESI+): m/z 432.1 (M+H)

Example 76: Synthesis of compound 342:

Experimental Procedures:

Preparation of compound 2:

A mixture of compound 1 (2 g, 9.2 mmol, 1 eq ), Pd(dppf)Cl2 (338.0 mg, 461.9 pmol, 0.1 eq ), TEA (2.8 g, 27.7 mmol, 3.9 mL, 3 eq) in 50 mL of methanol was degassed and purged with CO for 3 times, and then the mixture was stirred at 65°C for 12 hours under CO atmosphere (50 psi). Then it was partitioned between 30 mL of water and 150 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of brine, dried over anhydrous Na 2 S0 4 , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ethenethyl acetate =15:1 to 5: 1) to give 760 mg of compound 2 (3.9 mmol, 42.1% yield) as a white solid.

Preparation of compound 3:

To a solution of compound 2 (760 mg, 3.9 mmol, 1 eq ) in 10 mL of H2SO4 was added HNO3 (720.1 mg, 7.8 mmol, 514.3 pL, 68% purity, 2 eq) at 0°C. The mixture was stirred at 20°C for 5 hours. Then it was partitioned between 150 mL of saturated Na2CC>3 and 450 mL of ethyl acetate. The organic phase was separated, washed with 150 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether:ethyl acetate =1:0 to 10: 1) to give 760 mg of compound 3 (3.2 mmol, 81.3% yield) as a yellow solid.

Preparation of compound 4 :

To a solution of compound 3 (760 mg, 3.2 mmol, 1 eq) in 10 mL of ethanol and 1 mL of water was added Fe (529.2 mg, 9.5 mmol, 3 eq) and NH4CI (506.9 mg, 9.5 mmol, 3 eq). The mixture was stirred at 80°C for 2 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. Then it was partitioned between 10 mL of water and 50 mL of ethyl acetate. The organic phase was separated, washed with 25 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 200 mg of crude compound 4 as a yellow solid, which was used directly into the next step without purification.

Preparation of compound 5:

To a solution of compound 4 (200 mg, 949.6 pmol, 1 eq) in 3 mL of dichloromethane was added TEA (192.2 mg, 1.9 mmol, 264.3 pL, 2 eq) and 3-chloropropane-l-sulfonyl chloride

(252.2 mg, 1.4 mmol, 1.5 eq) at 0°C. The mixture was stirred at 20°C for 12 hours. The reaction mixture was concentrated under reduced pressure to give 330 mg of crude compound 5 as a yellow oil, which was used directly into the next step without purification.

Preparation of compound 6:

To a solution of compound 5 (330 mg, 939.6 pmol, 1 eq) in 5 mL of methanol was added TEA (142.6 mg, 1.4 mmol, 196.2 pL, 1.5 eq). The mixture was stirred at 65°C for 2 hours.

Then it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (S1O2, eluting with a gradient of petroleum ethenethyl acetate =10:1 to 1:3) to give 170 mg of compound 6 ( 540.1 pmol, 57.5% yield) as a yellow solid.

Preparation of compound? :

To a solution of compound 6 (70 mg, 222.4 pmol, 1 eq ) in 2 mL of methanol was added NaOH (17.8 mg, 444.8 pmol, 2 eq) and 0.5 mL of water. The mixture was stirred at 25°C for 1 hour. Then it was partitioned between 3 mL of IN HC1 and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 60 mg of compound 7 as a yellow solid, which was used directly into the next step without purification. Preparation of compound 342:

7 342

To a solution of compound 7 (60 mg, 199.5 pmol, 1 eq) in 2 mL of dimethyl formamide was added compound 8 (31.9 mg, 199.5 pmol, 1 eq) and TEA (30.3 mg, 299.3 pmol, 1.5 eq). The mixture was cooled to 0°C. The mixture was added HATU (113.8 mg, 299.3 pmol, 1.5 eq) and stirred at 20°C for 3 hours. Then it was partitioned between 3 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep- HPLC (TLA condition) to give 20.7 mg of compound 342 (46.7 m mol, 23.4% yield, 100% purity) as a white solid.

J H NMR (400MHz, DMSO-de) d ppm 10.99 (s, 1H), 7.93 (s, 1H), 7.78 (d, 7 =2.9 Hz, 1H), 7.75 (s, 1H), 7.68 (d, 7 =2.9 Hz, 1H), 7.47 (s, 1H), 3.82 (t, 7 =6.5 Hz, 2H), 3.61 (t, 7 =7.3 Hz, 2H), 2.59 (t, 7 =7.5 Hz, 2H), 2.43 (t, 7 =6.9 Hz, 2H), 1.61 - 1.53 (m, 2H), 0.87 (t, 7 =7.3 Hz, 3H)

LCMS (ESI+): m/z 442.9(M+H)

Example 77: Synthesis of compound 308:

5 308 Experimental Procedures:

Preparation of compound 2:

NaH (707.4 mg, 17.7 mmol, 60% purity, 2 eq) was added into a solution of 4-nitro-lH- pyrazole (1 g, 8.8 mmol, 1 eq ) in 20 mL of tetrahydrofuran at 0°C. Then the mixture was warmed to 25°C and stirred for 0.5 hour. A solution of SEM-C1 (1.8 g, 10.6 mmol, 1.9 mL, 1.2 eq) in 20 mL of tetrahydrofuran was added dropwise to the mixture at 25°C. The mixture was stirred for 1 hour. Then it was partitioned between 50 mL of water and 30 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 2 g of crude compound 2 as a yellow oil.

Preparation of compound 3:

To a solution of compound 2 (1 g, 4.1 mmol, 1 eq) and NH4CI (1.1 g, 20.6 mmol, 5 eq) in

20 mL of ethanol and 4 mL of water was added Le (1.1 g, 20.6 mmol, 5 eq). The mixture was stirred at 80°C for 0.5 hour. The mixture was filtered and the filtrate concentrated under reduced pressure to give a residue. Then it was partitioned between 50 mL of water and 60 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 870 mg of crude compound 3 as a red oil.

Preparation of compound 5:

To a solution of compound 3 (500 mg, 1.1 mmol, 1 eq ), compound 4 (459.7 mg, 2.2 mmol, 2 eq), Xantphos (93.5 mg, 161.6 pmol, 0.2 eq) and t-BuOK (241.8 mg, 2.2 mmol, 2 eq) in 4 mL of toluene was added Pd 2 (dba) 3 (98.6 mg, 107.7 pmol, 0.1 eq) under N2. The mixture was stirred at 110°C for 3 hours. The mixture was filtered and the filtrate concentrated under reduced pressure to give a residue. Then it was partitioned between 5 mL of water and 15 mL of ethyl acetate. The organic phase was separated, washed with 5 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (S1O2, eluting with a gradient of petroleum ether: ethyl acetate=l :0 to 1:2) to give 300 mg of compound 5 (502.8 pmol, 46.7% yield) as a brown oil.

Preparation of compound 308:

To a solution of compound 5 (50 mg, 83.8 pmol, 1 eq) in 0.5 mL of dichloromethane was added TFAA (88.0 mg, 419.0 pmol, 5 eq). The mixture was stirred at 25°C for 1 hour. The mixture was washed with 5 mL of sat. aqueous of NaHCCL and extracted with 15 mL of dichloromethane. The organic phase was separated, washed with 10 mL of brine, dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep- HPLC (basic condition) to give 1.4 mg of compound 308 (2.8 pmol, 3.4% yield, 94.2% purity) as a white solid.

J H NMR (400 MHz, DMSO-de) d ppm 12.76 - 12.55 (m, 1H), 10.51 - 10.45 (m, 1H), 7.85 - 7.78 (m, 1H), 7.59 - 7.51 (m, 1H), 7.38 - 7.24 (m, 2H), 7.17 - 7.04 (m, 2H), 6.51 - 6.44 (m, 1H), 3.81 - 3.74 (m, 2H), 3.59 - 3.53 (m, 2H), 2.43 - 2.41 (m, 2H).

LCMS (ESI+): m/z 466.1(M+H)

Example 78: In vitro efficacy assay of exemplary compounds

Exemplary compounds of the invention were evaluated for efficacy in inhibiting TDP-43 inclusions cellular imaging based assays. Two cell lines were developed utilizing mouse neuroblastoma N2a cells (ATCC, cat#: CCL-131). The first cell line expresses wild type human TDP-43 with a C-terminus eGFP tag. The second cell line expresses a mutant human TDP- 43::eGFP fusion protein in which the nuclear localization signal (NLS) of TDP-43 amino acids was mutated (K82A/R84A/K85A, K95A/K97A/K98A) (Winton, MJ, et al. (2007) JBC, 283: 13302-9). TDP-43WT: :eGFP and TDP-43ANLS::eGFP genes were synthesized and cloned into a cumate inducible PiggyBac vector (System Biosciences, cat. #: PBQM812A-1) and transfected into N2a cells using standard Lipofectamine 2000 protocol according to

manufacturer’s instruction (ThermoFisher). Puromycin resistance colonies were selected and further screened for expression by fluorescent microscopy and Western blot. The clones selected for TDP-43 aggregation assays are referred to as N2a TDP-43WT cells and N2a TDP- 43ANLS cells hereafter. While TDP-43WT is localized exclusively in nuclear, the defect of NLS signal in N2a TDP-43ANLS cells results in a largely cytoplasmic compartment.

The third cellular model is in human neuroblastoma SH SY5Y cells (ATCC, cat#: CRL- 2266). The parent cell line was first engineered to stably express a tetracycline repressor protein (designated as TREx-SY5Y cells, customer cell line development by ThermoFisher). Wild-type human TDP-43 with a C-terminus eGFP tag was synthesized and cloned into pcDNA5/TO expression vector (ThermoFisher). The plasmid was transfected into TREx-SY5Y cells using Lipofectamine 2000 and hygromycin resistant colonies were selected. Expression of TDP- 43WT::eGFP was examined under fluorescent microscopy and Western blot upon tetracycline induction. The clone that is used for TDP-43 aggregation assay is referred to as SY5Y TDP- 43WT cells hereafter.

Compound inhibition of TDP-43 aggregation was tested in an 8-point dose curve in 96- well format. Briefly, TDP-43 WT: :eGFP expression was induced with 15 mM cumate for 48 hrs (N2a TDP-43WT and N2a TDP43ANLS cells) or with 1 uM tetracycline for 24 hrs (SY5Y TDP- 43WT cells). The cells were then pre-treated for 1 hour with exemplary compounds before adding sodium arsenite to a final concentration of 15 mM and incubated for another 23 hrs. At the end of the treatment the cell monolayers were washed in PBS and fixed in 4%

paraformaldehyde (PFA), diluted from 16% stock (Electron Microscopy Sciences cat: #15710- S). The inhibitory effect on TDP-43 aggregation was measured using Celllnsight CX7 high content imager (ThermoFisher). The percentage of cells with TDP-43 aggregates was calculated based on the total number of cells identified by DAPI staining. An 8-point dose response curve was generated, and the ICso for each compound tested was determined and is summarized in Table 2 below. The compound numbers correspond to those shown in Figures 1A-1TT. In the table,“A” indicates an ICso of less than 100 nM,“B” indicates an IC50 range from 100 nM to 500 nM;“C” indicates an IC50 range from 500 nM to 2 mM; and“D” indicates an IC50 greater than 2 pM.

Table 2: Efficacy of exemplary compounds of the invention

EQUIVALENTS

It will be recognized that one or more features of any embodiments disclosed herein may be combined and/or rearranged within the scope of the invention to produce further

embodiments that are also within the scope of the invention.

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be within the scope of the present invention.

Although the invention has been described and illustrated in the foregoing illustrative embodiments, it is understood that the present disclosure has been made only by way of example, and that numerous changes in the details of implementation of the invention can be made without departing from the spirit and scope of the invention, which is limited only by the claims that follow. Features of the disclosed embodiments can be combined and/or rearranged in various ways within the scope and spirit of the invention to produce further embodiments that are also within the scope of the invention. Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments described specifically in this disclosure. Such equivalents are intended to be encompassed in the scope of the following claims.

All patents, patent applications and publications cited herein are hereby incorporated by reference in their entirety. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described and claimed herein.




 
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