FIELD OF THE INVENTION

The present invention relates to compounds inhibiting or otherwise modulating the activity of bromodomain-containing proteins, compositions and formulations comprising such compounds, and methods of using and preparing such compounds.

BACKGROUND OF THE INVENTION

Bromodomain (BRD) proteins containing BET (bromodomain and extra-terminal domain) family include four types: BRD2, BRD3, BRD4, and BRDT. Proteins of BET family are epigenetic coded readers that acetylate lysine residues on histones to alter chromatin structure and gene expression. BRD2, BRD3 and BRD4 are universally expressed, while BRDT is restricted to germ cells. BET protein plays a necessary but non-overlapping role in regulating gene transcription and controlling cell growth. BET proteins are associated with large protein complexes that regulate the transcription of many genes, including RNA polymerase II (Pol II) and forward transcription elongation factor (P-TEFb). It has been confirmed that BRD2 and BRD4 proteins maintain binding to chromosomes during mitosis and are required to promote transcription of important genes (including cyclin D and c-Myc) that initiate the cell cycle (Mochizuki, J Biol. Chem. 2008 283: 9040-9048). BRD4 protein is combined with RNA polymerase II (Pol II) and positive transcription elongation factor (P-TEFb) to jointly promote the transcription and expression of many genes related to cancer cell proliferation and apoptosis, such as c-Myc, cyclin, anti apoptosis protein Bcl-2, and regulate the growth and proliferation of tumor cells (Jang et al., Mol. Cell 2005 19:523-534). In some situations, the kinase activity of BRD4 can directly phosphorylate and activate RNA polymerase II (Devaiah et al., PNAS 2012 109: 6927-6932). Cells lacking BRD4 show impaired cell cycle progression. BRD2 and BRD3 have been reported to be associated with histones and actively transcribed genes and can be involved in promoting transcription elongation (Leroy et al., Mol. Cell. 2008 30: 51-60). In addition to acetylated histones, BET proteins have been shown to selectively bind acetylated transcription factors, including the RelA subunit of NF-kB and GATA1, thereby directly regulating the transcriptional activity of these proteins to control the expression of genes involved in inflammation and hematopoietic differentiation (Huang et al., Mol. Cell Biol. 2009 29: 1375-1387; Lamonica Proc. Nat. Acad. Sci. 2011 108: E159-168).

BET proteins including BRD4 have been identified as important mediators that alter gene expression characteristics found in a number of diseases including cancer, diabetes, obesity, atherosclerosis, cardiovascular, renal disease and viral infections. Read for reference Muller, S., et al., Expert Rev. Mol. Med., 13: e29 (2011); Zhou, M., et al., J. Virol., 83: 1036-1044 (2009); Chung, C W, et al., J. Med. Chem., 54: 3827-3838 (2011). For example, Myc is involved in most human cancers, and BET protein has been identified as a regulator of c-Myc; inhibition of BET protein (including BRD4) has been shown to down-regulate Myc transcription.

Therefore, there is a great need to develop compounds for use as bromodomain inhibitors. In particular, the development of compounds for use as BET inhibitors will be highly anticipated. Although it has been reported that some small molecule BET inhibitors have been used in clinical research, there are currently no drugs approved for marketing. Therefore, still need to develop new small molecule BET inhibitors for the clinical treatment of BET-mediated diseases or illness offers a new medication option.

SUMMARY OF INVENTION

The present invention relates to compounds as bromodomain inhibitors, especially BRD4 inhibitors, and their use in the treatment of BET-mediated diseases. The present invention first provides a compound shown in formula (I) or a pharmaceutically acceptable salt thereof,



Wherein,

R1and R2are each independently selected from H, C1-6alkyl, C1-6alkoxy, C6-10aryl or C5-10heteroaryl, the C1-6alkyl, C1-6alkoxy, C6-10aryl or C5-10heteroaryl is optionally substituted by C1-6alkyl, —NH2, —OH, C6-10aryl or C5-10heteroaryl; the C5-10heteroaryl has 1, 2, or 3 heteroatoms independently selected from nitrogen, oxygen or sulfur;

Q is absent or selected from C1-6alkylene, —SO2— or —NH—, the C1-6alkylene or —NH— isoptionally substituted by halogen, C1-6alkyl or C1-6alkoxy;

X is selected from H, C1-6alkyl, C6-10aryl or C5-10heteroaryl, the C1-6alkyl, C6-10aryl or C5-10heteroaryl is optionally substituted by halogen, halo C1-6alkyl, C1-6alkyl, C1-6alkoxy, C1-6alkylthio, C1-6alkoxycarbonyl or C1-6alkyl-SO2—;

Y is


wherein ring A is a 5 or 6 membered ring containing 0, 1, 2 or 3 heteroatoms independently selected from N, O or S; ring B is phenyl or C5-6heteroaryl containing 0, 1, 2 or 3 heteroatoms each independently selected from N, O or S.

R3and R4are absent or are each independently selected from H, halogen, hydroxyl, amino, C1-6alkyl, C1-6alkoxy, cyano, oxo, or —N(R5)—SO2—R6;

R5and R6are each independently selected from H, C1-6alkyl or C1-6halogenoalkyl.

As for the compounds shown in formula (I), the invention further provides some preferred technical schemes.

In some embodiments, R1is selected from H, C1-4alkyl, phenyl or C5-6heteroaryl, the C1-4alkyl, phenyl or C5-6heteroaryl is optionally substituted by C1-6alkyl, —NH2, phenyl, or C5-6heteroaryl; preferably, the heteroaryl has 1, 2, or 3 heteroatoms independently selected from nitrogen or sulfur.

In some embodiments, R1is H, —CH3,



In some embodiments, R1is H, —CH3,



In some embodiments, R2is H or C1-3alkyl.

In some embodiments, R2is H or —CH3.

In some embodiments, Q is absent or is selected from —CH2—,


—NH— or —SO2—.

In some embodiments, X is selected from H, C1-3alkyl or phenyl, the phenyl is unsubstituted or optionally substituted by halogen, halo C1-3alkyl, C1-3alkyl, C1-3alkoxy, C1-3alkylthio, C1-3alkoxycarbonyl, or C1-3alkyl-SO2—.

In some embodiments, X is selected from H, C1-3alkyl or phenyl, the phenyl is unsubstituted or optionally substituted by F, Cl, methyl, trifluoromethyl, methoxy, methylthio, methoxycarbonyl or methyl-SO2—.

In some embodiments, X is —CH3, H,



In some embodiments, Y is selected from


wherein,represents a single bond or a double bond, the U, V, W, G or Z is independently selected from C or N; when G is N, R4is absent; when G is C, R4is H or —N(R5)—SO2—R6; wherein, R5and R6are each independently selected from H, C1-6alkyl or C1-6halogenoalkyl.

In some embodiments, Y is selected from


wherein,represents a single bond or a double bond, the U, W or Z is independently selected from C or N; R3is selected from H, halogen, hydroxyl, amino, C1-6alkyl, C1-6alkoxy, cyano or oxo; R4is H or —N(R5)—SO2—R6; wherein R5and R6each are independently selected from H, C1-6alkyl or C1-6halogenoalkyl.

In some embodiments, R3is H, C1-6alkyl, C1-6alkoxy, cyano or oxo.

In some embodiments, R3is H, methyl or oxo.

In some embodiments, R4is —N(R5)—SO2—R6.

In some embodiments, R5and R6are independently selected from H or C1-6alkyl.

In some embodiments, R5and R6are independently selected from H, methyl or ethyl.

In some embodiments, Y is



Preferably, Y is


In the present invention, when Y is


it not only maintains better potent, but also has better PK properties.

The present invention further provides a compound or a pharmaceutically acceptable salt thereof, the compound refers to:1) 4-(1-(4-chlorobenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;2) 4-(1-(4-methoxybenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;3) 6-methyl-4-(2-methyl-1-(4-(methylthio)benzyl)-1H-imidazo[4,5-b]pyridin-6-yl)-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;4) 6-methyl-4-(2-methyl-1-(4-(trifluoromethyl)benzyl)-1H-imidazo[4,5-b]pyridin-6-yl)-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;5) 4-(1-(3-chlorobenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;6) 4-(1-benzyl-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyrid in-7(6H)one;7) 4-(1,2-dimethyl-1H-imidazo[4,5-b]pyridin-6-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;8) 6-methyl-4-(2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;9) methyl 4-((2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1H-imidazo[4,5-b]pyridin-1-yl)methyl)benzoate;10) 6-benzyl-4-(1-(4-methoxybenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;11) 6-isobutyl-4-(1-(4-methoxybenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;12) 6-ethyl-4-(1-(4-methoxybenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;13) 4-(1-(4-methoxybenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-2-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;14) 4-(1-(4-methoxybenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-6-(thiazol-2-ylmethyl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;15) 4-(1-(4-methoxybenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-6-(pyrazol-2-methyl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;16) 4-(1-(3-methoxybenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-2-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;17) 4-(1-(4-chlorobenzyl)-2-methyl-1H-imidazo[4,5-b]pyridin-6-yl)-6-(pyridin-3-ylmethyl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;18) 4-(1-(4-chlorobenzyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;19) 6-methyl-4-(2-methyl-1-(4-(trifluoromethyl)benzyl)-1H-imidazo[4,5-b]pyrazin-6-yl)-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;20) 4-(1-(4-methoxybenzyl)-2-methyl-H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrol o[2,3-c]pyridin-7(6H)-one;21) 4-(1-(1-(4-chlorophenyl)ethyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;22) 4-(1-benzyl-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyrid in-7(6H)-one;23) 4-(1-(3-trifluoromethylbenzyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;24) 4-(1-(2-fluoro-5-trifluoromethylbenzyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;25) 4-(1-(3-fluoro-5-trifluoromethylbenzyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;26) 4-(1-(2-fluoro-4-chlorobenzyl)-2-methyl-H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-H-pyrrolo[2,3-c]pyridin-7(6H)-one;27) 4-(1-(3-trifluoromethyl-4-chlorobenzyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;28) 4-(1-(3-fluoro-4-trifluoromethylbenzyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;29) 4-(1-(3-chloro-4-trifluoromethylbenzyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;30) 4-(1-(3-chlorobenzyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;31) 4-(1-(2,4-difluorobenzyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;32) 4-(1-(4-bromobenzyl)-2-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-6-methyl-H-pyrrolo[2,3-c]pyridin-7(6H)-one;33) 6-methyl-4-(2-methyl-1-(4-(methylsulfonyl)benzyl)-1H-imidazo[4,5-b]pyrazin-6-yl)-1H-pyrrolo[2,3-c]pyridin-7(6H)-one;34) 1-(4-chlorobenzyl)-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1,3-dihydro-2H-imidazo[4,5-b]pyridin-2-one;35) 4-(3-(1-(2,6-dichloro-3-fluorophenyl)ethyl)-2-methyl-H-pyrrolo[2,3-b]pyridin-5-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;36) 4-(1-(2,6-dichlorobenzyl)-1H-pyrrolo[3,2-b]pyridin-6-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;37) 4-(4-((4-chlorophenyl)amino)pyrido[2,3-d]pyrimidin-6-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;38) 4-(1-(2,6-dichlorobenzyl)-2-methyl-1H-pyrrolo[3,2-b]pyridin-6-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;39) 4-(1-(4-chlorobenzyl)-1H-pyrazolo[4,3-b]pyridin-6-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;40) 4-(1-((4-chlorophenyl)sulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one;41) N-(1-(4-chlorobenzyl)-2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;42) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(4-(trifluoromethyl)benzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;43) N-(1-(4-methoxybenzyl)-2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-H-pyrrolo[2,3-c]pyridin-4-yl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;44) N-(1-(1-(4-chlorophenyl)ethyl)-2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-H-pyrrolo[2,3-c]pyridin-4-yl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;45) N-(1-benzyl-2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;46) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(3-(trifluoromethyl)benzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;47) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(2-fluoro-5-(trifluoromethyl)benzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;48) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(3-fluoro-5-(trifluoromethyl)benzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;49) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(2-fluoro-4-chlorobenzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;50) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(3-(trifluoromethyl)-4chlorobenzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;51) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(3-fluoro-4-(trifluoromethyl)benzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;52) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(3-chloro-4-(trifluoromethyl)benzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;53) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(3-chlorobenzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;54) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(2,4-difluorobenzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;55) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(4-bromobenzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;56) N-(2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1-(4-(methylsulfonyl)benzyl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;57) N-(1-(2-chloro-4-fluorobenzyl)-2-methyl-6-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-1H-benzo[d]imidazol-4-yl)ethanesulfonamide;58) N-(5-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-3-(4-(trifluoromethyl)benzyl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide;59) N-(3-(2,4-difluorobenzyl)-5-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide;60) N-(3-(1-(4-chlorophenyl)ethyl)-5-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide;61) N-(3-(2-fluoro-5-(trifluoromethyl)benzyl)-5-(6-methyl-7-oxo-6,7-dihydro-H-pyrrolo[2,3-c]pyridin-4-yl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide;62) N-(3-(3,5-difluorobenzyl)-5-(6-methyl-7-oxo-6,7-dihydro-H-pyrrolo[2,3-c]pyridin-4-yl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide;63) N-(5-(6-methyl-7-oxo-6,7-dihydro-H-pyrrolo[2,3-c]pyridin-4-yl)-3-(2-(trifluoromethyl)benzyl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide;64) N-(3-(2,4-difluorobenzyl)-2-methyl-5-(6-methyl-7-oxo-6,7-dihydro-H-pyrrolo[2,3-c]pyridin-4-yl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide;65) N-(2-methyl-5-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-3-(4-(trifluoromethyl)benzyl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide;66) N-(2-methyl-5-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-3-(2-(trifluoromethyl)benzyl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide;67) N-(3-(3,5-difluorobenzyl)-2-methyl-5-(6-methyl-7-oxo-6,7-dihydro-H-pyrrolo[2,3-c]pyridin-4-yl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide; or68) N-(3-(2,6-dimethylbenzyl)-2-methyl-5-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-3H-imidazo[4,5-b]pyridin-7-yl)ethanesulfonamide.

The invention further provides crystalline form I of 6-methyl-4-(2-methyl-1-(4-(trifluoromethyl)benzyl)-1H-imidazo[4,5-b]pyrazin-6-yl)-1H-pyrrolo[2,3-c]pyridin-7(6H)-one (Compound 19).



In some embodiments, the X-ray powder diffraction pattern of the above crystalline form I has characteristic peaks with diffraction angles 2θ of 13.80.2°, 18.90.2°, 26.00.2°.

In some embodiments, the X-ray powder diffraction pattern of the above crystalline form I has characteristic peaks with diffraction angles 2θ of 6.2±0.2°, 13.8±0.2°, 18.9±0.2°, 19.5±0.2°, 26.0±0.2°, 26.8±0.2°.

In some embodiments, the above crystalline form I has an X-ray powder diffraction pattern as shown inFIG. 1.

The present invention summarized the characteristic peaks in the X-ray powder diffraction pattern of the above crystalline form I, as shown in Table 1.

TABLE 1Crystal planeRelative intensityNo.2θ ± 0.2 (°)spacing [Å](%)16.214.260.8213.86.4100.0318.94.731.6419.54.510.4526.03.431.8626.83.322.1

In some embodiments, crystalline form I of the present invention can be identified by differential scanning calorimetry. In some embodiments, crystalline form I has a differential scanning calorimetry curve as shown inFIG. 2. In the DSC pattern, the endothermic peak of crystalline form I is about 288.9° C. Differential scanning calorimetry analysis was performed by TA instruments Q200 DSC (purge gas: nitrogen; flow rate: 40 mL/min; heating rate: 10° C./min).

In some embodiments, the crystalline form I of the present invention can be identified by1HNMR, and the data of1HNMR is as follows:1H NMR (400 MHz, DMSO) δ 12.17 (s, 1H), 8.94 (s, 1H), 8.05 (s, 1H), 7.75 (d, J=8.0 Hz, 2H), 7.50 (d, J=8.0 Hz, 2H), 7.30 (m, 1H), 6.77 (m, 1H), 5.70 (s, 2H), 3.65 (s, 3H), 2.65 (s, 3H).

Preferably, the purity of the crystalline form I is ≥85%.

Preferably, the purity of the crystalline form I is ≥95%.

Preferably, the purity of the crystalline form I is ≥99%.

Preferably, the purity of the crystalline form I is ≥99.5%.

Preferably, the crystalline form I is anhydrous.

The crystalline form I provided by the invention has the characteristics of good crystallinity, non-hygroscopicity and good stability, and has acceptable oral bioavailability.

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the above compounds and a pharmaceutically acceptable excipient, such as hydroxypropyl methyl cellulose. In the composition, the said compound in a weight ratio to the said excipient within the range from about 0.0001 to about 10.

In addition, the present invention also provides a method for treating a subject suffering from a disease or disorder that responds to an inhibitory response to a bromodomain-containing protein, which comprises administering the therapeutically effective amount of the compound of formula (I) or the pharmaceutically acceptable salt thereof. In certain aspects, the bromodomain-containing protein is BRD4.

In certain aspects, the disease or disorder is selected from autoimmune diseases, inflammatory diseases, neurodegenerative diseases, cardiovascular disorders, kidney disorders, viral infections, and obesity. In certain aspects, the disease or disorder is selected from rheumatoid arthritis, osteoarthritis, atherosclerosis, psoriasis, systemic lupus erythematosus, multiple sclerosis, inflammatory bowel disease, asthma, chronic obstructive airway disease, pneumonia, dermatitis, hair loss, nephritis, vasculitis, atherosclerosis, Alzheimer's disease, hepatitis, primary biliary cirrhosis, sclerosing cholangitis, diabetes (including type 1 diabetes), acute rejection of transplanted organs. In certain aspects, the disease or disorder is cancer, including hematological cancer, lymphoma, multiple myeloma, leukemia, neoplasm, cancer, or tumor (eg, solid tumor). In certain aspects, the disease or disorder is tumor or cancer of colon, rectum, prostate (eg castrate resistant prostate cancer), lung cancer (eg non-small cell lung cancer and/or small cell lung cancer), pancreas, liver, kidney, cervix, uterus, stomach, ovary, breast (eg basal or basal-like breast cancer and/or triple negative breast cancer), skin (eg melanoma), nervous system (including brain, meninges, and central nervous system, Including neuroblastoma, glioblastoma, meningioma and medulloblastoma). In certain aspects, the disease or disorder is cancer. In certain aspects, the disease or disorder is hepatocellular carcinoma. In certain aspects, the disease or disorder is lymphoma. In certain aspects, the disease or disorder is B-cell lymphoma. In certain aspects, the disease or disorder is Burkitt's lymphoma. In certain aspects, the disease or disorder is diffuse large B-cell lymphoma. In certain aspects, the disease or disorder is multiple myeloma. In certain aspects, the disease or disorder is chronic lymphocytic leukemia. In certain aspects, the disease or disorder is NUT midline cardinoma. In certain aspects, the subject is human.

In certain aspects, the compound is administered intravenously, intramuscularly, parenterally, nasally, or orally. In one aspect, the compound is administered orally.

The present invention also provides a method for inhibiting bromodomain proteins, which comprises contacting the bromodomain proteins with a compound of formula (I) or a pharmaceutically acceptable salt thereof.

The present invention also provides the use of the compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a disease or condition that responds to the inhibition of bromodomain proteins.

The present invention also provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for treatment. Further provided is the compound of formula (I) or the pharmaceutically acceptable salt thereof for treating a subject suffering from a disease or disorder that responds to an inhibitory response to a bromodomain-containing protein. Also provided is the compound of formula (I) or pharmaceutically acceptable salts thereof for the above treatment methods.

In the present invention, unless otherwise stated, the term “halogen” refers to fluorine, chlorine, bromine or iodine. Preferably, halogen refers to fluorine, chlorine and bromine.

In the present invention, unless otherwise stated, the term “alkyl” includes a straight, branched or cyclic saturated monovalent hydrocarbon. For example, alkyl includes methyl, ethyl, propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, 3-(2-methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, cyclopentyl, n-hexyl, 2-hexyl, 2-methylpentyl and cyclohexyl. Similarly, “C1-6” in C1-6alkyl refers to a group containing 1, 2, 3, 4, 5 or 6 carbon atoms arranged in a linear or branched chain.

The alkoxy refers to oxyether formed from the above-mentioned straight chain, branch chain or cyclic alkyl.

In the present invention, unless otherwise stated, the term “heteroaryl” refers to a substituted or unsubstituted stable monocyclic or bicyclic group containing at least one aromatic ring of 5 to 10 ring atoms, the aromatic ring containing one, two or three ring heteroatoms selected from N, O and S, the remaining ring atoms are C atoms. Examples of such heteroaryl groups include, but are not limited to, pyridyl, pyrimidinyl, pyrrolyl, imidazolyl, thiazolyl, thienyl, benzimidazole.

The term “composition”, as used herein, is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combinations of the specified ingredients in the specified amounts. Accordingly, is pharmaceutical compositions containing the compounds of the present invention as the active ingredient as well as methods of preparing the instant compounds are also part of the present invention. Furthermore, some of the crystalline forms for the compounds may exist as polymorphs and as such are intended to be included in the present invention. In addition, some of the compounds may form solvates with water (i.e., hydrates) or common organic solvents and such solvates are also intended to be encompassed within the scope of this invention.

The present invention includes within its scope the prodrugs of the compounds of this invention. In general, such prodrugs will be functional derivatives of the compounds that are readily converted in vivo into the required compound. Thus, in the methods of treatment of the present invention, the term “administering” shall encompass the treatment of the various disorders described with the compound specifically disclosed or with a compound which may not be specifically disclosed, but which converts to the specified compound in vivo after administration to the subject. Conventional procedures for the selection and preparation of suitable prodrug derivatives has been described in bools such as “Design of Prodrugs”, ed. H. Bundgaard, Elsevier, 1985.

Obviously, the definition of any substituent or variable in a specific position in a molecule is independent of other positions in the molecule. It is easy to understand that a person of ordinary skill in the art can select a substituent or a substituted form of the compound of the present invention through the prior art means and the method described in the present invention to provide a chemically stable and easily synthesized compound.

The present invention includes compounds described herein can contain one or more asymmetric centers and may thus give rise to diastereomers and optical isomers. The present invention includes all such possible diastereomers as well as their racemic mixtures, their substantially pure resolved enantiomers, all possible geometric isomers, and pharmaceutically acceptable salts thereof.

The above Formula I are shown without a definitive stereochemistry at certain positions. The present invention includes all stereoisomers of Formula I and pharmaceutically acceptable salts thereof. Further, mixtures of stereoisomers as well as isolated specific stereoisomers are also included. During the course of the synthetic procedures used to prepare such compounds, or in using racemization or epimerization procedures known to those skilled in the art, the products of such procedures can be a mixture of stereoisomers.

When a tautomer of the compound of Formula I exists, the present invention includes any possible tautomers and pharmaceutically acceptable salts thereof, and mixtures thereof, except where specifically stated otherwise.

When the compound of Formula I and pharmaceutically acceptable salts thereof exist in the form of solvates or polymorphic forms, the present invention includes any possible solvates and polymorphic forms. A type of a solvent that forms the solvate is not particularly limited so long as the solvent is pharmacologically acceptable. For example, water, ethanol, propanol, acetone and similar solvents can be used.

The term “pharmaceutically acceptable salts” refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids. When the compound of the present invention is acidic, its corresponding salt can be conveniently prepared from pharmaceutically acceptable non-toxic bases, including inorganic bases and organic bases. Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (ic and ous), ferric, ferrous, lithium, magnesium, manganese (ic and ous), potassium, sodium, zinc and the like salts. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, as well as cyclic amines and substituted amines such as naturally occurring and synthesized substituted amines. Other pharmaceutically acceptable organic non-toxic bases from which salts can be formed include ion exchange resins such as, for example, arginine, betaine, caffeine, choline, N′,N′-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like.

When the compound of the present invention is basic, its corresponding salt can be conveniently prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include, for example, acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, formic, fumaric, gluconic, glutamic, hydrobromic, is hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid and the like. Preferred are citric, hydrobromic, formic, hydrochloric, maleic, phosphoric, sulfuric and tartaric acids, particularly preferred are formic and hydrochloric acid. Since the compounds of Formula I are intended for pharmaceutical use they are preferably provided in substantially pure form, for example at least 60% pure, more suitably at least 75% pure, especially at least 98% pure (% are on a weight for weight basis).

The pharmaceutical compositions of the present invention comprise a compound represented by Formula I (or a pharmaceutically acceptable salt thereof) as an active ingredient, a pharmaceutically acceptable carrier and optionally other therapeutic ingredients or adjuvants. Although the most suitable route in any given case will depend on the particular host to be administered, the nature of the host and severity of the conditions, the pharmaceutical of the present invention include compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration. The pharmaceutical compositions may be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

As described herein, the new crystalline forms may be identified by powder X-ray diffraction spectrum. However, persons skilled in the art know that the peak intensity and/or peak condition of powder X-ray diffraction may be different due to different experimental conditions, such as different diffraction test conditions and/or orientation priority. And because of the different accuracy of different instruments, the measured 2θ value will have an error of about ±0.2°. However, it is known that the relative strength value of the peak is more dependent on some properties of the measured sample than the position of the peak, such as the size of the crystal in the sample, the orientation of the crystal and the purity of the material to be analyzed, so the peak strength deviation shown can occur in the range of about 20% or more. However, despite the experimental error, instrument error, and orientation priority, persons skilled in the art can also obtain sufficient information to identify the crystalline form from the XRD data provided in the patent.

In practice, the compounds represented by Formula I, or a prodrug, or a metabolite, or pharmaceutically acceptable salts thereof, of this invention can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral or parenteral (including intravenous). Thus, the pharmaceutical compositions of the present invention can be presented as discrete units suitable for oral administration such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient. Further, the compositions can be presented as a powder, as granules, as a solution, as a suspension in an aqueous liquid, as a non-aqueous liquid, as oil-in-water emulsion, or as a water-in-oil liquid emulsion. In addition to the common dosage forms set out above, the compound represented by Formula I, or a pharmaceutically acceptable salt thereof, may also be administered by controlled release means and/or delivery devices. The compositions may be prepared by any of the methods of pharmacy. In general, such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped into the desired presentation.

Thus, the pharmaceutical compositions of this invention may include a pharmaceutically acceptable carrier and a compound, or a pharmaceutically acceptable salt, of Formula I. The compounds of Formula I, or pharmaceutically acceptable salts thereof, can also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds.

The pharmaceutical carrier employed can be, for example, a solid, liquid, or gas. Examples of solid carriers include such as lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples of liquid carriers include such as sugar syrup, peanut oil, olive oil, and water. Examples of gaseous carriers include such as carbon dioxide and nitrogen. In preparing the compositions for oral dosage form, any convenient pharmaceutical media may be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like may be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like may be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solid pharmaceutical carriers are employed. Optionally, tablets may be coated by standard aqueous or nonaqueous techniques.

A tablet containing the composition of this invention may be prepared by compression or molding, optionally with one or more accessory ingredients or adjuvants. Compressed tablets may be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, 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. Each tablet preferably contains from about 0.05 mg to about 5 g of the active ingredient and each cachet or capsule preferably containing from about 0.05 mg to about 5 g of the active ingredient. For example, a formulation intended for the oral administration to humans may contain from about 0.5 mg to about 5 g of active agent, compounded with an appropriate and convenient amount of carrier material which may vary from about 5 to about 95 percent of the total composition. Unit dosage forms will generally contain between from about 1 mg to about 2 g of the active ingredient, typically 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg.

Pharmaceutical compositions of the present invention suitable for parenteral administration may be prepared as solutions or suspensions of the active compounds in water. A suitable surfactant can be included such as, for example, hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof in oils. Further, a preservative can be included to prevent the detrimental growth of microorganisms.

Pharmaceutical compositions of the present invention suitable for injectable use include sterile aqueous solutions or dispersions. Furthermore, the compositions can be in the form of sterile powders for the extemporaneous preparation of such sterile injectable solutions or dispersions. In all cases, the final injectable form must be sterile and must be effectively fluid for easy syringability. The pharmaceutical compositions must be stable under the conditions of manufacture and storage; thus, preferably should be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.

Pharmaceutical compositions of the present invention can be in a form suitable for topical use such as, for example, an aerosol, cream, ointment, lotion, dusting powder, or the like. Further, the compositions can be in a form suitable for use in transdermal devices. These formulations may be prepared, utilizing a compound represented by Formula I of this invention, or a pharmaceutically acceptable salt thereof, via conventional processing methods. As an example, a cream or ointment is prepared by admixing hydrophilic material and water, together with about 5 wt % to about 10 wt % of the compound, to produce a cream or ointment having a desired consistency.

Pharmaceutical compositions of this invention can be in a form suitable for rectal administration with solid as a carrier. It is preferable that the mixture forms unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. The suppositories may be conveniently formed by first admixing the composition with the softened or melted carrier(s) followed by chilling and shaping in molds.

In addition to the aforementioned carrier ingredients, the pharmaceutical formulations described above may include, as appropriate, one or more additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, preservatives (including antioxidants) and the like. Furthermore, other adjuvants can be included to render the formulation isotonic with the blood of the intended recipient. Compositions containing a compound described by Formula I, or pharmaceutically acceptable salts thereof, may also be prepared in powder or liquid concentrate form.

Generally, dosage levels on the order of from about 0.01 mg/kg to about 150 mg/kg of body weight per day are useful in the treatment of the above-indicated conditions, or alternatively about 0.5 mg to about 7 g per patient per day. For example, colon cancer, rectal cancer, mantle cell lymphoma, multiple myeloma, breast cancer, prostate cancer, glioblastoma, squamous cell esophageal cancer, liposarcoma, T-cell lymphoma melanoma, pancreatic cancer, glioblastoma or lung cancer, may be effectively treated by the administration of from about 0.01 to 50 mg of the compound per kilogram of body weight per day, or alternatively about 0.5 mg to about 3.5 g per patient per day.

However, it is understood that the specific dose level for any particular patient will depend on a number of factors, including age, weight, general health, gender, diet, timing of administration, route of administration, excretion rate, status of combination medications and the severity of the specific disease being treated.

