BCAT Modulation

Abstract

The present disclosure relates, in part, to treating organic acidemias in a subject in need thereof by administering a therapeutically effective amount of a compound that inhibits BCAT2. The present disclosure also relates, in part, to methods for identifying candidate compounds for the treatment of organic acidemias. TIFF2022540336000314.tif113139

Description

[Technical Field]

[0001] Field The present disclosure relates to methods and compositions useful for targeting branched-chain amino acid aminotransferase (BCAT) for the treatment of inherited disorders of amino acid metabolism. [Background technology]

[0002] background Branched-chain organic aciduria and / or organic acidemia are a group of disorders resulting from defects in specific enzymes involved in the catabolism of the branched-chain amino acids (BCAAs) leucine, isoleucine, and valine. Organic acidemias are a subset of rare diseases affecting approximately 150,000 patients worldwide. In patients with organic acidemias, overproduction of metabolites occurs within mitochondria, resulting in the toxic accumulation of substrate molecules. Some of these toxic molecules negatively affect normal metabolic pathways by allosterically inhibiting critical enzymes. This effect on intracellular metabolism can lead to neurodevelopmental and developmental pathologies and, if untreated, death. Maple syrup urine disease (MSUD), isovaleric acidemia (IVA), propionic aciduria (PA), and methylmalonic aciduria (MMA) are the most common abnormal organic acidemias. However, any mutation in the BCAA metabolic pathway that results in the accumulation of metabolic intermediates may also be considered an organic acidemia. MMA, PA, and IVA present with severe dehydration, leuconeutropenia, and thrombocytopenia, and may mimic sepsis. These disorders can be diagnosed by newborn genetic screening and the identification of acylcarnitines and other organic acid compounds in plasma and urine by gas chromatography-mass spectrometry or tandem MS-MS.

[0003] The catabolism of BCAAs involves a common first step involving BCAA aminotransferase (BCAT) and branched-chain α-keto acid dehydrogenase (BCKD). Further metabolism utilizes distinct pathways, resulting in different end products (e.g., glucose and / or ketone bodies). However, the fact that the flux-generating step for the catabolism of the three BCAAs (involving BCKD) occurs at one of the common steps suggests that the generation of these downstream products may not be individually regulated and therefore may not play separate important roles. BCAA catabolism is highly regulated by both allosteric and covalent mechanisms. BCKD is inhibited by phosphorylation and activated by dephosphorylation. Allosteric inhibition of kinases by branched-chain keto acids (BCKAs), particularly by α-ketoisocaproate, serves both to promote the catabolism of excess amounts of these amino acids and as a mechanism for maintaining low concentrations of these dietary essential amino acids. Cytosolic and mitochondrial isoenzymes of BCAT have been identified.

[0004] At present, substrate level reduction is used to treat patients with various rare diseases.However, there is currently no available treatment for organic acidemia, such as MSUD, MMA, PA and IVA.Therefore, there is still a need for the therapeutic agent that is useful for treating these diseases and other genetic disorders of amino acid metabolism. Summary of the Invention

[0005] overview In some aspects, provided herein are methods of treating organic acidemias, comprising administering a therapeutically effective amount of a BCAT2 inhibitor compound described herein.

[0006] In some aspects, provided herein are methods of reducing toxic branched chain amino acid (BCAA) metabolites, comprising administering a therapeutically effective amount of a BCAT2 inhibitor compound described herein.

[0007] In some aspects, provided herein are methods of reducing intracellular toxic burden in a patient with organic acidemia, comprising administering a therapeutically effective amount of a BCAT2 inhibitor described herein.

[0008] In some embodiments, the compounds described herein have the formula (I): TIFF0007789565000001.tif43128 or a pharmaceutically acceptable salt thereof, During the ceremony, Z can be CH2, CF2, O or a bond; J 1 , J 2 , J 3 and J. 4 each may independently be CH or N; X 1 is N or CR 5A and; X 2 is N or CR 5B and; R 1 is C1-C6 alkyl, -NR 3 S(O)R 4 , -NR 3 SO2R 4 , -NR 3 C(O)R 4 , -C(O)(CH2) m NR 3 R 4 , C6~C 10 may be aryl, 5-10 membered heteroaryl or (5-10 membered heteroaryl)-C1-C6 alkyl; R 2 is -NR 6 R 7 , C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 It can be an aryl or a 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO2, C1-C6 alkyl, C1-C6 alkoxy, haloC1-C6 alkyl, haloC1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)NR 6R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC1-C6 alkyl), C6-C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' the substituents, together with the atom to which they are attached, may form a 5-membered heterocyclyl or a C3-C8 cycloalkyl, each optionally substituted with halo; R 3 can be H or C1-C6 alkyl; R 4 is H, C1-C6 alkyl, C2-C6 alkenyl, C6-C 10 It can be an aryl or a 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, C6-C 10 Aryl, -(CH2) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with; R 5A is H, halo, -CN, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, -(CH2) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 It can be; R5B is H, halo, -CN, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, -(CH2) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 Is it; Or R 5A and R 5B may be taken together with the atom to which they are attached to form a 6-membered heterocyclyl ring optionally substituted with oxo; R 6 and R 7 can independently be H, C1-C6 alkyl, or C3-C8 cycloalkyl; Each R 8 can independently be H, C1-C6 alkyl, or C3-C8 cycloalkyl; Each R 9 can independently be H, C1-C6 alkyl, C2-C6 alkenyl, or C3-C8 cycloalkyl; m can be 0 or 1; q can be 0 or 1; r can be 0 or 1.

[0009] In some embodiments, R 5A is not hydrogen.

[0010] In some embodiments, R 5B is hydrogen or -C(O)NR 8 R 9 No.

[0011] In some embodiments, R 2 is not unsubstituted aryl or unsubstituted heteroaryl. 2 C6-C substituted with one -S-C1-C6 alkyl 10 In some embodiments, R is not aryl. 2 is not a 5-10 membered heteroaryl substituted with one C1-C6 alkyl.

[0012] In some embodiments, the compound of formula (I) is TIFF0007789565000002.tif102147TIFF0007789565000003.tif202146TIFF0007789565000004.tif101147 and pharmaceutically acceptable salts and stereoisomers thereof is not selected from the group consisting of:

[0013] In some embodiments, Z can be CH. In other embodiments, Z can be CF. In still other embodiments, Z can be O. In still other embodiments, Z can be a bond.

[0014] In some embodiments, J 1 can be CH. In other embodiments, J 1 can be N.

[0015] In some embodiments, J 2 can be CH. In other embodiments, J 2 can be N.

[0016] In some embodiments, J 3 is CH. In other embodiments, J 3 can be N.

[0017] In some embodiments, J 4 can be CH. In other embodiments, J 4 can be N.

[0018] In some embodiments, X 1 can be N. In other embodiments, X 1 is CR 5A It could be.

[0019] In some embodiments, X 2 can be N. In other embodiments, X 2 is CR 5B It could be.

[0020] In some embodiments, J2 can be N, and X 1 can be N. In other embodiments, J 2 can be N, and J 3 can be N. In other embodiments, J 2 can be N, and J 4 can be N. In still other embodiments, J 2 can be N, and X 2 can be N. In some embodiments, J 2 , J 3 and J. 4 Each of X may be CH; 1 is CR 5A X 2 is CR 5B It could be.

[0021] In some embodiments, R 1 NR 3 C(O)R 4 In other embodiments, R 1 -C(O)(CH2) m NR 3 R 4 It could be.

[0022] In some embodiments, R 3 can be hydrogen. In other embodiments, R 3 C 1~6 It can be alkyl.

[0023] In some embodiments, R 4 each containing one, two or three substituents R 4' C optionally substituted with 6~10 aryl or 5-10 membered heteroaryl. In some embodiments, R 4 may be selected from phenyl, thiophene, oxazole, isoxazole, thiazole, furan and pyrrole, each of which may contain one, two or three substituents R 4' where each R 4'may be independently selected from the group consisting of -F, -Cl, -Br, -CN, -NO2, -CF3, -CH3, -CH(CH3)2, -C(O)CH3, -C(O)OCH3, phenyl, cyclopropyl, and morpholinyl.

[0024] In some embodiments, m can be 0. In other embodiments, m can be 1.

[0025] In some embodiments, R 2 each containing one, two or three substituents R 2' C optionally substituted with 6~10 aryl or 5-10 membered heteroaryl. In some embodiments, R 2 may be selected from the group consisting of phenyl, pyridine, pyrimidine, pyridazine, pyrazine, imidazole or thiophene, each of which may contain one, two or three substituents R 2' where each R 2' may be independently selected from the group consisting of -F, -Cl, -Br, -CN, NO, -CH, -CFH, -C≡CH, -C(O)H, -CONH, -C(O)NHCH, -OH, -OCH, -OCF, -SCF, -NH, -NHC(O)CH, and morpholine.

[0026] In some embodiments, X 2 is CR 5B R 5B -C(O)NR 8 R 9 In other embodiments, X 2 is CR 5B R 5B halo, -CN, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, -(CH2) r OH, -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 In yet another embodiment, X 2 is CR 5B R 5B Ga-NR8 COR 9 In some embodiments, X 2 is CR 5B R 5B -C(O)OR 8 In some embodiments, X 2 is CR 5B R 5B can be -CF3, -COCH3, -CH2OH, -CN or tetrazole.

[0027] In some embodiments, R 8 can be hydrogen. In other embodiments, R 8 Ga-C 1~6 It can be alkyl.

[0028] In some embodiments, R 9 can be hydrogen. In other embodiments, R 9 Ga-C 1~6 It can be alkyl.

[0029] In some embodiments, the compounds provided herein have the formula (Ia): or a pharmaceutically acceptable salt thereof, During the ceremony, R 1 is C1-C6 alkyl, -NR 3 S(O)R 4 , -NR 3 SO2R 4 , -NR 3 C(O)R 4 , -C(O)(CH2) m NR 3 R 4 , C6~C 10 may be aryl or 5-10 membered heteroaryl; R 2 is C1-C6 alkyl, C6-C 10 may be aryl or 5-10 membered heteroaryl; R 4 is C6~C 10may be aryl or 5-10 membered heteroaryl; R 5B is -C(O)NR 8 R 9 It can be; R 8 and R 9 are independently H, C1-C6 alkyl, or C3-C8 cycloalkyl.

[0030] In some aspects, the term "aromatic group" is used herein to refer to a group comprising a compound of formula: R 1 Ga-NR 3 SO2R 4 or -NR 3 C(O)R 4 It can be; R 2 can be aryl or 5-10 membered heteroaryl; R 3 can be H or C1-C6 alkyl; R 4 C6~C 10 may be aryl or 5-10 membered heteroaryl; R 5B -C(O)NR 8 R 9 It can be; R 8 and R 9 can independently be H or C1-C6 alkyl Provided is a compound of formula (Ia) or a pharmaceutically acceptable salt thereof:

[0031] In some aspects, the term "aromatic group" is used herein to refer to a group comprising a compound of formula: R 1 can be -NR3C(O)R4; R 2 C6~C 10 may be aryl or 5-10 membered heteroaryl; R 3 can be H or methyl; R 4 C6~C 10 may be aryl or 5-10 membered heteroaryl; R 5B -C(O)NR8 R 9 It can be; R 8 and R 9 can independently be H or C1-C6 alkyl Provided is a compound of formula (Ia) or a pharmaceutically acceptable salt thereof:

[0032] In some aspects, the term "aromatic group" is used herein to refer to a group comprising a compound of formula: R1 can be -NR3C(O)R4; R 2 C6~C 10 may be aryl or 5-10 membered heteroaryl; R 3 can be H or methyl; R 4 C6~C 10 may be aryl or 5-10 membered heteroaryl; R 5 -C(O)NR 8 R 9 It can be; R 8 R 9 and may be, independently, H or methyl. Provided is a compound of formula (Ia) or a pharmaceutically acceptable salt thereof:

[0033] In some aspects, the compounds of the present disclosure are 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(2-(methylthio)phenyl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-phenyl-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-3-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-4-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiophen-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiophen-3-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(2-methylthiazol-4-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiazol-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(1H-pyrazol-3-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-cyanothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-chlorothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-methylthiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromo-N-methylthiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-((5-bromothiophene)-2-sulfonamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 2-bromo-N-(3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)thiazole-5-carboxamide, 1-(3-(5-bromo-1-methyl-1H-pyrrole-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(4-bromo-1H-pyrrole-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 3-chloro-N-(3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)isoxazole-5-carboxamide, 5-bromo-N-(3-(2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)thiophene-2-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N,N-dimethyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-cyclopropyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, and 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-isopropyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide; or a pharmaceutically acceptable salt thereof It could be.

[0034] In a further aspect, there is provided herein a compound of formula (II): There may be provided a compound of formula (I) having the structure of TIFF0007789565000006.tif48128, or a pharmaceutically acceptable salt thereof, wherein: R 2 is C6~C 10 It can be an aryl or a 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO2, C1-C6 alkyl, C1-C6 alkoxy, haloC1-C6 alkyl, haloC1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC1-C6 alkyl), C6-C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' the substituents, together with the atom to which they are attached, may form a 5-membered heterocyclyl or a C3-C8 cycloalkyl, each optionally substituted with halo; R 4 is halo, -CN, -NO2, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, C6-C 10 Aryl, -(CH2) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be a 5-10 membered heteroaryl optionally substituted with R 5B is -C(O)NR 8 R 9 It could be.

[0035] In some embodiments, R 2 is one, two, or three substituents R independently selected from halo, —CN, —NO2, and C1-C6 alkyl; 2' In other embodiments, R 2 is one, two, or three substituents R independently selected from halo, —CN, —NO2, and C1-C6 alkyl; 2' and 5-10 membered heteroaryl optionally substituted with

[0036] In some embodiments, R 4 may be a thiophene optionally substituted with halo or phenyl.

[0037] In a further aspect, the compounds provided herein have the formula (III): or a pharmaceutically acceptable salt thereof, During the ceremony, each J 3 and J. 4 are independently CH or N; X 1 is N or CR 5A It can be; X 2 is N or CR 5B It can be; Here, J 3 , J 4 , X 1 and X 2 of which 1, 2, 3 or 4 are N; R 2 is C6~C10 It can be an aryl or a 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO2, C1-C6 alkyl, C1-C6 alkoxy, haloC1-C6 alkyl, haloC1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC1-C6 alkyl), C6-C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' the substituents, together with the atom to which they are attached, form a 5-membered heterocyclyl or C3-C8 cycloalkyl, each optionally substituted with halo; R 4 is H, C1-C6 alkyl, C2-C6 alkenyl, C6-C 10 It can be an aryl or a 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, C6-C 10 Aryl, -(CH2) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with.

[0038] In some embodiments, R 2 is one, two, or three substituents R independently selected from halo, —CN, —NO2, and C1-C6 alkyl; 2'In other embodiments, R 2 is one, two, or three substituents R independently selected from halo, —CN, —NO2, and C1-C6 alkyl; 2' and 5-10 membered heteroaryl optionally substituted with

[0039] In some embodiments, R 4 may be a thiophene optionally substituted with halo or phenyl.

[0040] In some embodiments, X 2 is CR 5B R 5B -H, -halo, -C(O)NR 8 R 9 , -NR 8 COR 9 or -C(O)OR 8 R 8 and R 9 may each independently be H or C1-C6 alkyl.

[0041] In a further aspect, the compounds provided herein have the formula (IV): or a pharmaceutically acceptable salt thereof, wherein: R 1 is C1-C6 alkyl, -NR 3 C(O)R 4 , -C(O)(CH2) m NR 3 R 4 , C6~C 10 aryl, 5-10 membered heteroaryl, or (5-10 membered heteroaryl)-C1-C6 alkyl; R 2 Ha-NR 6 R 7 , C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10aryl or 5-10 membered heteroaryl, wherein each aryl or heteroaryl is halo, -CN, -NO2, C1-C6 alkoxy, haloC1-C6 alkyl, haloC1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, C6~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, form a 5-membered heterocyclyl or C3-C8 cycloalkyl, each optionally substituted with halo.

[0042] In still a further aspect, there is provided herein a compound of formula (V): There may be provided a compound of formula (I) having the structure: TIFF0007789565000009.tif44128, or a pharmaceutically acceptable salt thereof: During the ceremony, R 5A is halo, -CN, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, -(CH2) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 and; R 5B is H, halo, -CN, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, -(CH2) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR8 Is it; Or R 5A and R 5B may be taken together with the atom to which they are attached to form a 6-membered heterocyclyl ring which may be substituted by oxo.

[0043] In still yet a further aspect, there is provided herein a compound of formula (VI): There may be provided a compound of formula (I) having the structure TIFF0007789565000010.tif44128, or a pharmaceutically acceptable salt thereof.

[0044] In a still yet further aspect, there is provided herein a compound of formula (VII): There may be provided a compound of formula (I) having the structure TIFF0007789565000011.tif46128, or a pharmaceutically acceptable salt thereof.

[0045] In still yet a further aspect, provided herein: TIFF0007789565000012.tif181152TIFF0007789565000013.tif181146TIFF0007789565000014.tif205155TIFF00077 89565000015.tif216153TIFF0007789565000016.tif207155TIFF0007789565000017.tif192149TIFF00077895650000 18.tif193148TIFF0007789565000019.tif197146TIFF0007789565000020.tif189148TIFF0007789565000021.tif192 146TIFF0007789565000022.tif193147TIFF0007789565000023.tif184145TIFF0007789565000024.tif187149TIFF000 7789565000025.tif181144TIFF0007789565000026.tif193145TIFF0007789565000027.tif185145TIFF000778956500 0028.tif184144TIFF0007789565000029.tif188146TIFF0007789565000030.tif182147TIFF0007789565000031.tif1 79145TIFF0007789565000032.tif180147TIFF0007789565000033.tif184152TIFF0007789565000034.tif181147TIFF0007789565000035.tif134144TIFF0007789565000036.tif187142TIFF0007789565000037.tif90147 and pharmaceutically acceptable salts thereof are provided.

[0046] In some embodiments, the compounds disclosed herein selectively inhibit BCAT2 relative to BCAT1.

[0047] In some aspects, the methods described herein reduce and / or ameliorate the symptoms of organic acidemia in a patient.

[0048] In some aspects, the organic acidemia is any form of methylmalonic acidemia (MMA), any form of propionic acidemia (PA), isovaleric acidemia, glutaric aciduria type 1 (GA1), beta-ketothiolase deficiency (BKT), 3-methylcrotonyl-CoA carboxylase deficiency (3-MCC), 3-hydroxy-3-methylglutaric-CoA lyase deficiency (HMG), 3-methylglutaconic acidemia, or 3-methylglutaconyl-CoA hydratase deficiency (M GA), D-2 hydroxyglutaric aciduria (D2-HGA), isobutyryl-CoA dehydrogenase deficiency 3-hydroxyisobutyric aciduria (ICBD), L-2-hydroxy-glutaric aciduria (L2HGA), malonyl-CoA decarboxylase deficiency, also known as malonic acidemia (MA), multiple carboxylase deficiency (MCD, holocarboxylase synthase) and 3-hydroxyisobutyryl-CoA hydrolase deficiency (HIBCH).

[0049] In some embodiments, the methods described herein cause a reduction in the level of toxic branched-chain amino acid (BCAA) metabolites. In some embodiments, the toxic metabolites are selected from propionic acid, 2-methylcitric acid, 3-hydroxypropionic acid, propionyl-CoA, methylmalonic acid, methylmalonyl-CoA, and isovaleric acid. In some embodiments, the toxic metabolites allosterically inhibit enzymes.

[0050] In some embodiments, the methods described herein result in increased levels of circulating branched chain amino acids (BCAAs), hi some embodiments, the branched chain amino acids are selected from one or more of leucine, isoleucine, and valine.

[0051] In some embodiments, the method described herein modulates the metabolic flux of BCAA pathway.In some embodiments, the metabolic flux is reduced before and / or after treatment.In some embodiments, the method described herein inhibits and / or reduces the BCAT2-mediated flux of BCAA pathway.

[0052] In some embodiments, the reduction in BCAT2-mediated flux occurs when leucine metabolism is reduced or inhibited. In some embodiments, the metabolism of leucine is reduced and / or inhibited in one or more enzymes in the BCAA metabolic pathway selected from branched-chain α-keto acid dehydrogenase (BCKDH), isovaleryl-CoA dehydrogenase (IVD), 3-methylcrotonyl-CoA carboxylase (3MCC), 3-methylglutaconic-CoA hydratase (3MGA), 3-hydroxy-3-methylglutaric-CoA lyase (HMGL), acetyl-CoA carboxylase (AC), and malonyl-CoA decarboxylase (MA).

[0053] In some embodiments, the reduction in BCAT2-mediated flux occurs when isoleucine metabolism is reduced and / or inhibited. In some embodiments, isoleucine metabolism is reduced and / or inhibited in one or more enzymes in the BCAA metabolic pathway selected from BCKDH, methylbutyryl-CoA dehydrogenase (SBCAD), hydratase, 2-methyl-3-hydroxyisobutyrate dehydrogenase (MHBD), acetoacetyl-CoA thiolase (T2), AC, MA, propionyl-CoA carboxylase (PCC), methylmalonyl-CoA mutase (MUT), and succinyl-CoA ligase (SUCLA).

[0054] In some embodiments, the reduction in BCAT2-mediated flux occurs when valine metabolism is reduced and / or abrogated in one or more enzymes in the BCAA metabolic pathway selected from BCKDH, isobutyryl-CoA dehydrogenase (IBDH), hydratase, 3-hydroxyisobutyryl-CoA deacylase (hydrolase) (HIBDA), 3-hydroxyisobutyrate dehydrogenase (HIBDH), methylmalonate semialdehyde dehydrogenase (MMSDH), PCC, MUT, and SUCLA.

[0055] In some embodiments, the methods described herein can further comprise administering an inhibitor of an amino acid transporter, optionally an SLC6A19 inhibitor. In certain embodiments, the level of branched-chain amino acids is reduced compared to treatment without the administration of the inhibitor of an amino acid transporter.

[0056] In yet another aspect, provided herein: (a) obtaining a test compound having the ability to bind directly or indirectly to BCAT2; (b) assaying for functional modulation of BCAT2; and (c) classifying the test compound as a candidate compound if decreased, low, or substantially no BCAT2 activity is detected. Methods for identifying candidate compounds for the treatment of organic acidemias are described, comprising:

[0057] In some embodiments, the functional modulation of BCAT2 is measured using cell-based assay.In some embodiments, when the increase of branched-chain amino acid is measured, the test compound is classified as candidate compound.In some embodiments, when the decrease of propionyl-carnitine is measured, the test compound is classified as candidate compound.

[0058] In other embodiments, the functional modulation of BCAT2 is measured using cell-free assay.In some embodiments, cell-free assay is enzyme-coupled fluorescent assay.In some embodiments, when measuring the decrease of L-glutamate, test compound is classified as candidate compound.In some embodiments, when measuring the decrease of α-ketoglutarate, test compound is classified as candidate compound.

[0059] In another aspect, provided herein: (a)(i) obtaining a test compound having the ability to bind directly or indirectly to BCAT2; (ii) assaying for functional modulation of BCAT2; and (iii) classifying the test compound as a candidate compound if a decrease in BCAT2 activity, low BCAT2 activity, or substantially no BCAT2 activity is detected. identifying a candidate compound; and (b) formulating the candidate compound for the treatment of organic acidemia The present invention provides a method for making a medicament for the treatment of organic acidemia, comprising:

[0060] In some embodiments, the functional modulation of BCAT2 is measured using cell-based assay.In some embodiments, when the increase of branched-chain amino acid is measured, the test compound is classified as candidate compound.In some embodiments, when the decrease of propionyl-carnitine is measured, the test compound is classified as candidate compound.

[0061] In other embodiments, the functional modulation of BCAT2 is measured using cell-free assay.In some embodiments, cell-free assay is enzyme-coupled fluorescent assay.In some embodiments, when measuring the decrease of L-glutamate, test compound is classified as candidate compound.In some embodiments, when measuring the decrease of α-ketoglutarate, test compound is classified as candidate compound.

[0062] In some aspects, the organic acidemia is any form of methylmalonic acidemia (MMA), any form of propionic acidemia (PA), isovaleric acidemia, glutaric aciduria type 1 (GA1), beta-ketothiolase deficiency (BKT), 3-methylcrotonyl-CoA carboxylase deficiency (3-MCC), 3-hydroxy-3-methylglutaric-CoA lyase deficiency (HMG), 3-methylglutaconic acidemia, or 3-methylglutaconyl-CoA hydratase deficiency (MG). A), D-2 hydroxyglutaric aciduria (D2-HGA), isobutyryl-CoA dehydrogenase deficiency 3-hydroxyisobutyric aciduria (ICBD), L-2-hydroxy-glutaric aciduria (L2HGA), malonyl-CoA decarboxylase deficiency, also known as malonic acidemia (MA), multiple carboxylase deficiency (MCD, holocarboxylase synthase) and 3-hydroxyisobutyryl-CoA hydrolase deficiency (HIBCH).

[0063] [The present invention 1001] 1. A method of treating organic acidemia, comprising administering a therapeutically effective amount of a BCAT2 inhibitor compound of formula (I) or a pharmaceutically acceptable salt thereof: TIFF0007789565000038.tif44128 During the ceremony, Z is CH 2 , C.F. 2 , O or a bond; J 1 、J 2 、J 3 and J. 4 each is independently CH or N; X 1 is N or CR 5A and; X 2 is N or CR 5B and; R 1 is C 1 ~C 6 Alkyl, -NR 3 S(O)R4 , -NR 3 SO 2 R 4 , -NR 3 C(O)R 4 , -C(O)(CH 2 ) m NR 3 R 4 、C 6 ~C 10 Aryl, 5- to 10-membered heteroaryl or (5- to 10-membered heteroaryl)-C 1 ~C 6 is alkyl; R 2 Ha-NR 6 R 7 、C 1 ~C 6 Alkyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC 1 ~C 6 alkyl), C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 forming a cycloalkyl; R 3 is H or C 1 ~C 6 is alkyl; R 4 is H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 Aryl, -(CH 2 ) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with; R 5A H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 and; R 5B H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 Is it; Or R 5A and R 5B together with the atoms to which they are attached form a 6-membered heterocyclyl ring which may be substituted with oxo; R 6 and R 7 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 is cycloalkyl; Each R 8 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 is cycloalkyl; Each R 9 are independently H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl or C 3 ~C 8 is cycloalkyl; m is 0 or 1; q is 0 or 1; r is 0 or 1. [The present invention 1002] 1. A method of reducing toxic branched chain amino acid (BCAA) metabolites, comprising administering a therapeutically effective amount of a BCAT2 inhibitor compound of Formula I, or a pharmaceutically acceptable salt thereof: TIFF0007789565000039.tif44128 During the ceremony, Z is CH 2 , C.F. 2 , O or a bond; J 1 、J 2 、J 3 and J. 4 each is independently CH or N; X 1 is N or CR 5A and; X 2 is N or CR 5B and; R 1 is C 1 ~C 6 Alkyl, -NR 3 S(O)R 4 , -NR 3 SO 2 R 4 , -NR 3 C(O)R 4 , -C(O)(CH 2 ) m NR 3 R 4 、C 6 ~C 10 Aryl, 5- to 10-membered heteroaryl or (5- to 10-membered heteroaryl)-C 1 ~C 6 is alkyl; R 2 Ha-NR 6 R 7 、C 1 ~C 6 Alkyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC 1 ~C 6 alkyl), C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 forming a cycloalkyl; R 3 is H or C 1 ~C 6 is alkyl; R 4 is H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 Aryl, -(CH 2 ) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with; R 5A H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 and; R 5B H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 Is it; Or R 5A and R 5B together with the atoms to which they are attached form a 6-membered heterocyclyl ring which may be substituted with oxo; R 6 and R 7 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 is cycloalkyl; Each R 8 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 is cycloalkyl; Each R 9 are independently H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl or C 3 ~C 8 is cycloalkyl; m is 0 or 1; q is 0 or 1; r is 0 or 1. [The present invention 1003] 1. A method of reducing intracellular toxic burden in a patient having organic acidemia, comprising administering a therapeutically effective amount of a BCAT2 inhibitor compound of formula (I) or a pharmaceutically acceptable salt thereof: TIFF0007789565000040.tif44128 During the ceremony, Z is CH 2 , C.F. 2 , O or a bond; J 1 、J 2 、J 3 and J. 4 each is independently CH or N; X 1 is N or CR 5A and; X 2 is N or CR 5B and; R 1 is C 1 ~C 6 Alkyl, -NR 3 S(O)R 4 , -NR 3 SO 2 R 4 , -NR 3 C(O)R 4 , -C(O)(CH 2 )m NR 3 R 4 、C 6 ~C 10 Aryl, 5- to 10-membered heteroaryl or (5- to 10-membered heteroaryl)-C 1 ~C 6 is alkyl; R 2 Ha-NR 6 R 7 、C 1 ~C 6 Alkyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC 1 ~C 6 alkyl), C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 forming a cycloalkyl; R 3 is H or C 1 ~C 6 is alkyl; R 4 is H, C 1 ~C6 Alkyl, C 2 ~C 6 Alkenyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 Aryl, -(CH 2 ) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with; R 5A H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 and; R 5B H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 Is it; Or R 5A and R 5B together with the atoms to which they are attached form a 6-membered heterocyclyl ring which may be substituted with oxo; R 6 and R 7 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 is cycloalkyl; Each R 8 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 is cycloalkyl; Each R 9 are independently H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl or C 3 ~C 8 is cycloalkyl; m is 0 or 1; q is 0 or 1; r is 0 or 1. [The present invention 1004] Z is CH 2 or CF 2 The method of any one of inventions 1001 to 1003, [The present invention 1005] The method of any one of claims 1001 to 1003, wherein Z is O. [The present invention 1006] The method of any one of claims 1001 to 1003, wherein Z is a bond. [The present invention 1007] J 1 The method according to any one of claims 1001 to 1006, wherein is CH. [The present invention 1008] J 1 The method according to any one of claims 1001 to 1006, wherein is N. [The present invention 1009] J 2 The method according to any one of claims 1001 to 1008, wherein is CH. [The present invention 1010] J 2 The method according to any one of claims 1001 to 1008, wherein is N. [The present invention 1011] J 3 The method according to any one of claims 1001 to 1010, wherein is CH. [The present invention 1012] J 3 The method according to any one of claims 1001 to 1010, wherein is N. [The present invention 1013] J 4 The method according to any one of claims 1001 to 1012, wherein is CH. [The present invention 1014] J 4 The method of any one of claims 1001 to 1012, wherein is N. [The present invention 1015] X 1 The method according to any one of claims 1001 to 1014, wherein is N. [The present invention 1016] X 1 is CR 5A The method of any one of claims 1001 to 1014, [The present invention 1017] X 2 The method according to any one of claims 1001 to 1016, wherein is N. [The present invention 1018] X 2 is CR 5B The method of any one of claims 1001 to 1016, [The present invention 1019] J 2 is N and X 1 The method according to any one of claims 1001 to 1006, wherein is N. [The present invention 1020] J 2 is N and J 3 The method according to any one of claims 1001 to 1006, wherein is N. [The present invention 1021] J 2 is N and J 4 The method according to any one of claims 1001 to 1006, wherein is N. [The present invention 1022] J 2 is N and X 2 The method according to any one of claims 1001 to 1006, wherein is N. [The present invention 1023] J 2 、J 3 and J. 4 Each of the following is CH; 1 is CR 5A and X 2 is CR 5B The method of any one of inventions 1001 to 1006, [The present invention 1024] R 1 Ga-NR 3 C(O)R 4 or -C(O)(CH 2 ) m NR 3 R 4 Any one of the methods of the present inventions 1001 to 1023, [The present invention 1025] R 3 is hydrogen or C 1~6 The method of claim 1024, wherein the alkyl group is alkyl. [The present invention 1026] R 4 each containing one, two or three substituents R 4' C optionally substituted with 6~10 The method of any one of claims 1024 to 1025, wherein the aryl is aryl or 5- to 10-membered heteroaryl. [The present invention 1027] R 4 is selected from phenyl, thiophene, oxazole, isoxazole, thiazole, furan and pyrrole, each of which may contain one, two or three substituents R 4' where each R 4' -F, -Cl, -Br, -CN, -NO 2 , -CF 3 , -CH 3 , -CH(CH 3 ) 2 , -C(O)CH 3 , -C(O)OCH 3 , phenyl, cyclopropyl, and morpholinyl. [The present invention 1028] The method of any one of claims 1001 to 1027, wherein m is 0. [The present invention 1029] The method of any one of claims 1001 to 1027, wherein m is 1. [The present invention 1030] R 2 each containing one, two or three substituents R 2' C optionally substituted with 6~10 The method of any one of claims 1001 to 1029, wherein the aryl is aryl or 5- to 10-membered heteroaryl. [The present invention 1031] R 2 is selected from the group consisting of phenyl, pyridine, pyrimidine, pyridazine, pyrazine, imidazole or thiophene, each of which may contain one, two or three substituents R 2' where each R 2' -F, -Cl, -Br, -CN, NO 2 , -CH 3 , -CF 2 H, -C≡CH, -C(O)H, -CONH 2 , -C(O)NHCH 3 , -OH, -OCH 3 , -OCF 3 , -SCF 3 , -NH 2 , -NHC(O)CH 3 1030. The method of claim 1030, wherein the aryl group is independently selected from the group consisting of: and morpholine. [The present invention 1032] X 1 is CR 5A and R 5A But hydrogen, halo, C 1 ~C 6 Alkyl or C 1 ~C 6 10. The method of any one of claims 1001 to 1014, 1016 to 1018, or 1020 to 1031, wherein the alkyl group is selected from the group consisting of alkoxy. [The present invention 1033] R 5A -F, -Cl, -Br, -CH 3 , -OH and -OCH 3 The method of the present invention 1032, selected from the group consisting of: [The present invention 1034] X 2 is CR 5B and R 5 -C(O)NR 8 R 9 The method of any one of claims 1001 to 1016, 1018 to 1021, or 1023 to 1033 of the present invention. [This invention 1035] X 2 is CR 5B and R 5B Ga-NR 8 COR 9 or -C(O)OR 8 The method of any one of claims 1001 to 1016, 1018 to 1021, or 1023 to 1033 of the present invention. [The present invention 1036] R 8 is hydrogen or -C 1~6 The method of any one of claims 1034 to 1035, wherein the alkyl is alkyl. [This invention 1037] R 9 The method according to any one of claims 1034 to 1036, wherein is hydrogen. [The present invention 1038] R 9 Ga-C 1~6 The method of any one of claims 1034 to 1036, wherein the alkyl is alkyl. [This invention 1039] X 2 is CR 5B and R 5B Ga-CF 3 , -COCH 3 , -CH 2 The method of any one of claims 1001 to 1016, 1018 to 1021, or 1023 to 1033, wherein the compound is —OH, —CN, or tetrazole. [The present invention 1040] The method of any one of claims 1001 to 1003, wherein the compound of formula I is a compound of formula (Ia): TIFF0007789565000041.tif35128 During the ceremony, R 1 is C 1 ~C 6 Alkyl, -NR 3 S(O)R 4 , -NR 3 SO 2 R 4 , -NR 3 C(O)R 4 , -C(O)(CH 2 ) m NR 3 R 4 、C 6 ~C 10 aryl or 5-10 membered heteroaryl; R 2 is C 1 ~C 6 Alkyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl; R 4 is C 6 ~C 10 aryl or 5-10 membered heteroaryl; R 5B is -C(O)NR 8 R 9 and; R 8 and R 9 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 It is cycloalkyl. [The present invention 1041] R 1 Ga-NR 3 SO 2 R 4 or -NR 3 C(O)R 4 and; R 2 is aryl or 5-10 membered heteroaryl; R 3 is H or C 1 ~C 6 is alkyl; R 4 C 6 ~C 10 aryl or 5-10 membered heteroaryl; R 5B -C(O)NR 8 R 9 and; R 8 and R 9 are independently H or C 1 ~C 6 is alkyl, The method of the present invention 1038. [The present invention 1042] R 1 Ga-NR 3 C(O)R 4 and; R が C 6 ~C 10 aryl or 5-10 membered heteroaryl; R 3 is H or methyl; R 4 C 6 ~C 10 aryl or 5-10 membered heteroaryl; R 5B -C(O)NR 8 R 9 and; R 8 and R 9 are independently H or C 1 ~C 6 is alkyl, The method of the present invention 1038. [This invention 1043] R 1 Ga-NR 3 C(O)R 4 and; R 2 C 6 ~C 10 aryl or 5-10 membered heteroaryl; R 3 is H or methyl; R 4 C 6 ~C 10 aryl or 5-10 membered heteroaryl; R 5 -C(O)NR 8 R 9 and; R 8 and R 9 is independently H or methyl; The method of the present invention 1038. [This invention 1044] BCAT2 inhibitors 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(2-(methylthio)phenyl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-phenyl-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-3-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-4-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiophen-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiophen-3-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(2-methylthiazol-4-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiazol-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(1H-pyrazol-3-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-cyanothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-chlorothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-methylthiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromo-N-methylthiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-((5-bromothiophene)-2-sulfonamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 2-bromo-N-(3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)thiazole-5-carboxamide, 1-(3-(5-bromo-1-methyl-1H-pyrrole-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(4-bromo-1H-pyrrole-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 3-chloro-N-(3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)isoxazole-5-carboxamide, 5-bromo-N-(3-(2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)thiophene-2-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N,N-dimethyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-cyclopropyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, and 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-isopropyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide The method of any one of claims 1001 to 1003, wherein the compound is selected from the group consisting of: [This invention 1045] 1. A method of treating organic acidemias comprising administering a therapeutically effective amount of a BCAT2 inhibitor compound, wherein the compound: TIFF0007789565000042.tif185153TIFF0007789565000043.tif202153TIFF0007789565000044.tif190154TIFF0007789565000045.tif228154TIFF0007789565000046.tif211154TIFF0007789565000047.tif238151TIFF0007789565000048.tif214149TIFF0007789565000049.tif201148TIFF0007789565000050.tif196149TIFF0007789565000051.tif205148TIFF0007789565000052.tif195149TIFF0007789565000053.tif237147TIFF0007789565000054.tif202151TIFF0007789565000055.tif236145TIFF0007789565000056.tif202147TIFF0007789565000057.tif230147TIFF0007789565000058.tif206145TIFF0007789565000059.tif234148TIFF0007789565000060.tif233147TIFF0007789565000061.tif234150TIFF0007789565000062.tif195153TIFF0007789565000063.tif237145TIFF0007789565000064.tif236144TIFF0007789565000065.tif93149 and pharmaceutically acceptable salts thereof A method selected from the group consisting of: [The present invention 1046] 1. A method of reducing toxic branched chain amino acid (BCAA) metabolites, comprising administering a therapeutically effective amount of a BCAT2 inhibitor compound, wherein the compound: TIFF0007789565000066.tif93146TIFF0007789565000067.tif191154TIFF0007789565000068.tif236153TIFF0007789565000069.tif220154TIFF0007789565000070.tif217154TIFF0007789565000071.tif197153TIFF0007789565000072.tif196151TIFF0007789565000073.tif199150TIFF0007789565000074.tif202147TIFF0007789565000075.tif196150TIFF0007789565000076.tif198147TIFF0007789565000077.tif195149TIFF0007789565000078.tif191149TIFF0007789565000079.tif236147TIFF0007789565000080.tif193145TIFF0007789565000081.tif194147TIFF0007789565000082.tif236147TIFF0007789565000083.tif197146TIFF0007789565000084.tif235149TIFF0007789565000085.tif232149TIFF0007789565000086.tif236153TIFF0007789565000087.tif234153TIFF0007789565000088.tif231147TIFF0007789565000089.tif194149TIFF0007789565000090.tif45144 and pharmaceutically acceptable salts thereof A method selected from the group consisting of: [This invention 1047] 1. A method of reducing intracellular toxic burden in a patient having organic acidemia, comprising administering a therapeutically effective amount of a BCAT2 inhibitor compound, the compound comprising: TIFF0007789565000091.tif141145TIFF0007789565000092.tif234153TIFF0007789565000093.tif191154TIFF0007789565000094.tif237155TIFF0007789565000095.tif210154TIFF0007789565000096.tif196150TIFF0007789565000097.tif195151TIFF0007789565000098.tif196147TIFF0007789565000099.tif203148TIFF0007789565000100.tif199150TIFF0007789565000101.tif195149TIFF0007789565000102.tif195145TIFF0007789565000103.tif196151TIFF0007789565000104.tif234145TIFF0007789565000105.tif196147TIFF0007789565000106.tif193145TIFF0007789565000107.tif237147TIFF0007789565000108.tif193148TIFF0007789565000109.tif235149TIFF0007789565000110.tif232149TIFF0007789565000111.tif236154TIFF0007789565000112.tif234149TIFF0007789565000113.tif232147TIFF0007789565000114.tif189149 and pharmaceutically acceptable salts thereof A method selected from the group consisting of: [This invention 1048] The method of any of claims 1001 or 1004 to 1047, wherein the symptoms of organic acidemia in the patient are reduced and / or alleviated. [This invention 1049] Organic acidemias include any form of methylmalonic acidemia (MMA), any form of propionic acidemia (PA), isovaleric acidemia, glutaric aciduria type 1 (GA1), beta-ketothiolase deficiency (BKT), 3-methylcrotonyl-CoA carboxylase deficiency (3-MCC), 3-hydroxy-3-methylglutaric-CoA lyase deficiency (HMG), 3-methylglutaconic acidemia or 3-methylglutaconyl-CoA hydratase deficiency (MGA), D-2 hydrochloride, and 3-methylglutaconyl-CoA hydratase deficiency (D-2 hydrochloride). 2-hydroxy-glutaric aciduria (D2-HGA), isobutyryl-CoA dehydrogenase deficiency 3-hydroxyisobutyric aciduria (ICBD), L-2-hydroxy-glutaric aciduria (L2HGA), malonyl-CoA decarboxylase deficiency, also known as malonic acidemia (MA), multiple carboxylase deficiency (MCD, holocarboxylase synthase) and 3-hydroxyisobutyryl-CoA hydrolase deficiency (HIBCH). [The present invention 1050] The method of any of claims 1001 to 1049, wherein a reduction in the level of toxic branched chain amino acid (BCAA) metabolites is caused. [This invention 1051] 1050. The method of claim 1050, wherein the toxic metabolite is selected from propionic acid, 2-methylcitric acid, 3-hydroxypropionic acid, propionyl-CoA, methylmalonic acid, methylmalonyl-CoA, and isovaleric acid. [This invention 1052] The method of claim 1051, wherein the toxic metabolite allosterically inhibits the enzyme. [This invention 1053] The method of any of claims 1001 to 1052, wherein an increase in circulating branched chain amino acid (BCAA) levels is caused. [This invention 1054] The method of claim 1053, wherein the branched chain amino acids are selected from one or more of leucine, isoleucine, and valine. [This invention 1055] The method of any of claims 1001 to 1054, wherein metabolic flux through the BCAA pathway is modulated. [The present invention 1056] The method of claim 1055, wherein the reduction in metabolic flux occurs before and / or after treatment. [This invention 1057] The method of any of claims 1001 to 1056, wherein BCAT2-mediated flux through the BCAA pathway is inhibited and / or reduced. [This invention 1058] The method of claim 1057, wherein the reduction in BCAT2-mediated flux occurs when leucine metabolism is reduced or inhibited. [This invention 1059] 1058. The method of claim 1058, wherein the metabolism of leucine is reduced and / or abrogated in one or more enzymes in the BCAA metabolic pathway selected from branched-chain alpha-keto acid dehydrogenase (BCKDH), isovaleryl-CoA dehydrogenase (IVD), 3-methylcrotonyl-CoA carboxylase (3MCC), 3-methylglutaconic-CoA hydratase (3MGA), 3-hydroxy-3-methylglutaric-CoA lyase (HMGL), acetyl-CoA carboxylase (AC), and malonyl-CoA decarboxylase (MA). [The present invention 1060] The method of claim 1057, wherein the reduction in BCAT2-mediated flux occurs when isoleucine metabolism is reduced and / or inhibited. [This invention 1061] 1060. The method of claim 1060, wherein the metabolism of isoleucine is reduced and / or abrogated in one or more enzymes in the BCAA metabolic pathway selected from BCKDH, methylbutyryl-CoA dehydrogenase (SBCAD), hydratase, 2-methyl-3-hydroxyisobutyrate dehydrogenase (MHBD), acetoacetyl-CoA thiolase (T2), AC, MA, propionyl-CoA carboxylase (PCC), methylmalonyl-CoA mutase (MUT), and succinyl-CoA ligase (SUCLA). [This invention 1062] The method of claim 1057, wherein the reduction in BCAT2-mediated flux occurs when valine metabolism is reduced and / or inhibited. [This invention 1063] 1062. The method of claim 1062, wherein the metabolism of valine is reduced and / or abrogated in one or more enzymes in the BCAA metabolic pathway selected from BCKDH, isobutyryl-CoA dehydrogenase (IBDH), hydratase, 3-hydroxyisobutyryl-CoA deacylase (hydrolase) (HIBDA), 3-hydroxyisobutyrate dehydrogenase (HIBDH), methylmalonate semialdehyde dehydrogenase (MMSDH), PCC, MUT, and SUCLA. [This invention 1064] The method of any of claims 1001 to 1063, further comprising the step of administering an inhibitor of an amino acid transporter, optionally an SLC6A19 inhibitor. [This invention 1065] The method of claim 1064, wherein the level of branched chain amino acids is reduced compared to treatment without administration of an inhibitor of an amino acid transporter. [The present invention 1066] (a) obtaining a test compound having the ability to bind directly or indirectly to BCAT2; (b) assaying for functional modulation of BCAT2; and (c) classifying the test compound as a candidate compound if decreased, low, or substantially no BCAT2 activity is detected. 10. A method for identifying a candidate compound for the treatment of organic acidemia, comprising: [This invention 1067] The method of claim 1066, wherein the functional modulation of BCAT2 is assayed using a cell-based assay. [The present invention 1068] The method of claim 1067, wherein the test compound is classified as a candidate compound if an increase in branched chain amino acids is measured. [This invention 1069] The method of claim 1067, wherein the test compound is classified as a candidate compound if a decrease in propionyl-carnitine is measured. [The present invention 1070] The method of claim 1066, wherein the functional modulation of BCAT2 is assayed using a cell-free assay. [This invention 1071] 1070. The method of claim 1070, wherein the cell-free assay is an enzyme-coupled fluorescent assay. [This invention 1072] The method of any one of claims 1070 to 1071, wherein the test compound is classified as a candidate compound if a decrease in L-glutamate is measured. [This invention 1073] The method of any one of claims 1070 to 1071, wherein the test compound is classified as a candidate compound if a decrease in alpha-ketoglutarate is measured. [This invention 1074] (a)(i) obtaining a test compound having the ability to bind directly or indirectly to BCAT2; (ii) assaying for functional modulation of BCAT2; and (iii) classifying the test compound as a candidate compound if a decrease in BCAT2 activity, low BCAT2 activity, or substantially no BCAT2 activity is detected. identifying a candidate compound; and (b) formulating the candidate compound for the treatment of organic acidemia 10. A method for making a medicament for the treatment of organic acidemia, comprising: [This invention 1075] The method of claim 1074, wherein the functional modulation of BCAT2 is assayed using a cell-based assay. [This invention 1076] The method of claim 1074, wherein the test compound is classified as a candidate compound if an increase in branched chain amino acids is measured. [This invention 1077] The method of claim 1075, wherein the test compound is classified as a candidate compound if a decrease in propionyl-carnitine is measured. [This invention 1078] The method of claim 1074, wherein the functional modulation of BCAT2 is assayed using a cell-free assay. [This invention 1079] 1078. The method of claim 1078, wherein the cell-free assay is an enzyme-coupled fluorescent assay. [The present invention 1080] The method of any one of claims 1078 to 1079, wherein the test compound is classified as a candidate compound if a decrease in L-glutamate is measured. [This invention 1081] The method of any one of claims 1078 to 1079, wherein the test compound is classified as a candidate compound if a decrease in alpha-ketoglutarate is measured. [This invention 1082] Organic acidemias include any form of methylmalonic acidemia (MMA), any form of propionic acidemia (PA), isovaleric acidemia, glutaric aciduria type 1 (GA1), beta-ketothiolase deficiency (BKT), 3-methylcrotonyl-CoA carboxylase deficiency (3-MCC), 3-hydroxy-3-methylglutaric-CoA lyase deficiency (HMG), 3-methylglutaconic acidemia or 3-methylglutaconyl-CoA hydratase deficiency (MGA), D-2 hydroxyglutarate any of the methods of claims 1066 to 1081, wherein the malonyl-CoA decarboxylase deficiency is selected from the group consisting of malonic aciduria (D2-HGA), isobutyryl-CoA dehydrogenase deficiency, 3-hydroxyisobutyric aciduria (ICBD), L-2-hydroxy-glutaric aciduria (L2HGA), malonyl-CoA decarboxylase deficiency, also known as malonic acidemia (MA), multiple carboxylase deficiency (MCD, holocarboxylase synthase) and 3-hydroxyisobutyryl-CoA hydrolase deficiency (HIBCH). [This invention 1083] A compound of formula I or a pharmaceutically acceptable salt thereof: TIFF0007789565000115.tif44128 (In the formula, Z is CH 2 , C.F. 2 , O or a bond; J 1、J 2 、J 3 and J. 4 each is independently CH or N; X 1 is N or CR 5A and; X 2 is N or CR 5B and; R 1 is C 1 ~C 6 Alkyl, -NR 3 S(O)R 4 , -NR 3 SO 2 R 4 , -NR 3 C(O)R 4 , -C(O)(CH 2 ) m NR 3 R 4 、C 6 ~C 10 Aryl, 5- to 10-membered heteroaryl or (5- to 10-membered heteroaryl)-C 1 ~C 6 is alkyl; R 2 Ha-NR 6 R 7 、C 1 ~C 6 Alkyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC 1 ~C 6 alkyl), C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 forming a cycloalkyl; R 3 is H or C 1 ~C 6 is alkyl; R 4 is H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 Aryl, -(CH 2 ) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with; R 5A H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 and; R 5B H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 Is it; Or R 5A and R 5B together with the atoms to which they are attached form a 6-membered heterocyclyl ring which may be substituted with oxo; R 6 and R 7 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 is cycloalkyl; Each R 8 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 is cycloalkyl; Each R 9 are independently H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl or C 3 ~C 8 is cycloalkyl; m is 0 or 1; q is 0 or 1; (r is 0 or 1) However, the compound is TIFF0007789565000116.tif99154TIFF0007789565000117.tif241154TIFF0007789565000118.tif53152 and pharmaceutically acceptable salts and stereoisomers thereof not selected from the group consisting of A compound or a pharmaceutically acceptable salt thereof. [This invention 1084] Z is CH 2 or CF 2 The compound of the present invention 1083, [This invention 1085] The compound of the present invention 1083, wherein Z is O. [The present invention 1086] The compound of claim 1083, wherein Z is a bond. [This invention 1087] J 1 The compound of any one of claims 1083 to 1086, wherein is CH. [This invention 1088] J 1 The compound of any one of claims 1083 to 1086, wherein is N. [This invention 1089] J 2 The compound of any one of claims 1083 to 1088, wherein is CH. [The present invention 1090] J 2 The compound of any one of 1083 to 1088 of the present invention, wherein is N. [This invention 1091] J 3 The compound of any one of claims 1083 to 1090, wherein is CH. [This invention 1092] J 3 The compound of any one of claims 1083 to 1090, wherein is N. [This invention 1093] J 4 The compound of any one of claims 1083 to 1092, wherein is CH. [This invention 1094] J 4 The compound of any one of claims 1083 to 1092, wherein is N. [This invention 1095] X 1 The compound of any one of claims 1083 to 1094, wherein is N. [This invention 1096] X 1 is CR 5A The compound of any one of claims 1083 to 1094 of the present invention, [This invention 1097] X 2 The compound of any one of claims 1083 to 1096, wherein is N. [This invention 1098] X 2 is CR 5B The compound of any one of claims 1083 to 1096 of the present invention, [This invention 1099] J 2 is N and X 1 The compound of any one of claims 1083 to 1086, wherein is N. [The present invention 1100] J 2 is N and J 3 The compound of any one of claims 1083 to 1086, wherein is N. [The present invention 1101] J 2 is N and J 4 The compound of any one of claims 1083 to 1086, wherein is N. [The present invention 1102] J 2 is N and X 2 The compound of any one of claims 1083 to 1086, wherein is N. [The present invention 1103] J 2 、J 3 and J. 4 Each of the following is CH; 1 is CR 5A and X 2 is CR 5B The compound of any one of 1083 to 1086 of the present invention, [The present invention 1104] R 1 NR 3 C(O)R 4 or -C(O)(CH 2 ) m NR 3 R 4 The compound of any one of 1083 to 1103 of the present invention, [This invention 1105] R 3 is hydrogen or C 1~6 The compound of the present invention 1104, which is alkyl. [The present invention 1106] R 4 each containing one, two or three substituents R 4' C optionally substituted with 6~10 The compound of the present invention 1104 or 1105, which is aryl or 5- to 10-membered heteroaryl. [This invention 1107] R 4 is selected from phenyl, thiophene, oxazole, isoxazole, thiazole, furan and pyrrole, each of which may contain one, two or three substituents R 4' where each R 4' -F, -Cl, -Br, -CN, -NO 2 , -CF 3 , -CH 3 , -CH(CH 3 ) 2 , -C(O)CH 3 , -C(O)OCH 3 1106. A compound of the present invention, wherein the compound is independently selected from the group consisting of phenyl, cyclopropyl, and morpholinyl. [This invention 1108] The compound of any one of 1083 to 1107 of the present invention, wherein m is 0. [This invention 1109] The compound of any one of 1083 to 1107 of the present invention, wherein m is 1. [The present invention 1110] R 2 each containing one, two or three substituents R 2' C optionally substituted with 6~10 The compound of any one of claims 1083 to 1109, which is aryl or 5- to 10-membered heteroaryl. [The present invention 1111] R 2 is selected from the group consisting of phenyl, pyridine, pyrimidine, pyridazine, pyrazine, imidazole or thiophene, each of which may contain one, two or three substituents R 2' where each R 2' -F, -Cl, -Br, -CN, NO 2 , -CH 3 , -CF 2 H, -C≡CH, -C(O)H, -CONH 2 , -C(O)NHCH 3 , -OH, -OCH 3 , -OCF 3 , -SCF 3 , -NH 2 , -NHC(O)CH 3 and morpholine. [The present invention 1112] X 2 is CR 5B and R 5B -C(O)NR 8 R 9 The compound of any one of 1083 to 1096, 1098 to 1101, or 1103 to 1111 of the present invention, [The present invention 1113] X 2 is CR 5B and R 5B Halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 The compound of any one of 1083 to 1096, 1098 to 1101, or 1103 to 1111 of the present invention, [This invention 1114] X 2 is CR 5B and R 5B Ga-NR 8 COR 9 or -C(O)OR 8 The compound of the present invention 1113, [This invention 1115] R 8 is hydrogen or -C 1~6 The compound of the present invention 1113 or 1114, which is alkyl. [The present invention 1116] R 9 The compound of any one of 1113 to 1115 of the present invention, wherein is hydrogen. [This invention 1117] R 9 Ga-C 1~6 The compound of any one of 1113 to 1115 of the present invention, which is alkyl. [This invention 1118] X 2 is CR 5B and R 5B Ga-CF 3 , -COCH 3 , -CH 2 Compounds of any one of claims 1083 to 1096, 1098 to 1101 or 1103 to 1111, which are OH, -CN or tetrazole. [This invention 1119] The compound of the present invention has the structure of formula (II) or a pharmaceutically acceptable salt thereof: TIFF0007789565000119.tif50128 During the ceremony, R 2 is C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC 1 ~C 6 alkyl), C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 forming a cycloalkyl; R 4 Halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 Aryl, -(CH 2 ) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' is a 5-10 membered heteroaryl optionally substituted with R 5B is -C(O)NR 8 R 9 is. [The present invention 1120] R 2 But, halo, -CN, -NO 2 and C 1 ~C 6 one, two or three substituents R independently selected from alkyl 2' 1119 compounds of the present invention, wherein the phenyl is optionally substituted with . [This invention 1121] R 2 But, halo, -CN, -NO 2 and C 1 ~C 6 one, two or three substituents R independently selected from alkyl 2' The compound of the present invention 1119, wherein the compound is a 5- to 10-membered heteroaryl optionally substituted with [This invention 1122] R 4 is a thiophene optionally substituted with halo or phenyl. [This invention 1123] The compound of the present invention has the structure of formula (III) or a pharmaceutically acceptable salt thereof: TIFF0007789565000120.tif51128 During the ceremony, each J 3 and J. 4 are independently CH or N; X 1 is N or CR 5A and; X 2 is N or CR 5B and; Here, J 3 、J 4 、X 1 and X 2 of which 1, 2, 3 or 4 are N; R 2 is C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC 1 ~C 6 alkyl), C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 forming a cycloalkyl; R 4 is H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 Aryl, -(CH 2 ) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with. [This invention 1124] R 2 But, halo, -CN, -NO 2 and C 1 ~C 6 one, two or three substituents R independently selected from alkyl 2' The compound of the present invention 1123, which is phenyl optionally substituted by. [Invention 1125] R 2 But, halo, -CN, -NO 2 and C 1 ~C 6 one, two or three substituents R independently selected from alkyl 2' The compound of the present invention 1123, wherein the compound is a 5- to 10-membered heteroaryl optionally substituted with [The present invention 1126] R 4 The compound of any one of 1123 to 1125 of the present invention, wherein is thiophene optionally substituted with halo or phenyl. [This invention 1127] X 2 is CR 5B and; R 5B -H, -halo, -C(O)NR 8 R 9 , -NR 8 COR 9 or -C(O)OR 8 Selected from; R 8 and R 9 are each independently H or C 1 ~C 6 is alkyl, The compound of any one of 1123 to 1126 of the present invention. [This invention 1128] The compound of the present invention has the structure of formula (IV) or a pharmaceutically acceptable salt thereof: TIFF0007789565000121.tif45128 During the ceremony, R 1 is C 1 ~C 6 Alkyl, -NR 3 C(O)R 4 , -C(O)(CH 2 ) m NR3 R 4 、C 6 ~C 10 Aryl, 5- to 10-membered heteroaryl or (5- to 10-membered heteroaryl)-C 1 ~C 6 is alkyl; R 2 Ha-NR 6 R 7 、C 1 ~C 6 Alkyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 It forms a cycloalkyl. [This invention 1129] The compound of the present invention has the structure of Formula (V) or a pharmaceutically acceptable salt thereof: TIFF0007789565000122.tif45128 During the ceremony, R 1 is C 1 ~C 6 Alkyl, -NR 3 S(O)R 4 , -NR 3 SO 2 R 4 , -NR 3 C(O)R 4 , -C(O)(CH 2 ) m NR 3 R 4 、C 6 ~C 10 Aryl, 5- to 10-membered heteroaryl or (5- to 10-membered heteroaryl)-C 1 ~C 6 is alkyl; R 2 Ha-NR 6 R 7 、C 1 ~C 6 Alkyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 、C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC 1 ~C 6 alkyl), C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 forming a cycloalkyl; R 5A is halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 and; R 5B H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 Is it; Or R 5A and R 5B are taken together with the atoms to which they are attached to form a 6-membered heterocyclyl ring which may be substituted with oxo. [The present invention 1130] The compound of the present invention has the structure of formula (VI) or a pharmaceutically acceptable salt thereof: TIFF0007789565000123.tif46128 。 [This invention 1131] The compound of the present invention has the structure of formula (VII) or a pharmaceutically acceptable salt thereof: TIFF0007789565000124.tif47128 。 [This invention 1132] below: TIFF0007789565000125.tif198154TIFF0007789565000126.tif237153TIFF0007789565000127.tif22 3155TIFF0007789565000128.tif210154TIFF0007789565000129.tif206155TIFF0007789565000130.t if195150TIFF0007789565000131.tif200150TIFF0007789565000132.tif202147TIFF00077895650001 33.tif196150TIFF0007789565000134.tif198147TIFF0007789565000135.tif231149TIFF00077895650 00136.tif205151TIFF0007789565000137.tif234145TIFF0007789565000138.tif196147TIFF0007789 565000139.tif193145TIFF0007789565000140.tif237147TIFF0007789565000141.tif193148TIFF000 7789565000142.tif235149TIFF0007789565000143.tif232149TIFF0007789565000144.tif236154TIF F0007789565000145.tif234149TIFF0007789565000146.tif232147TIFF0007789565000147.tif189149 A compound selected from the group consisting of: or a pharmaceutically acceptable salt thereof. Details of the present disclosure are set forth in the accompanying description below. Although methods and materials similar or equivalent to those described herein can be used in practicing or testing the present disclosure, exemplary methods and materials are now described. Other features, objects, and advantages of the present disclosure will be apparent from the description and claims. In this specification and the appended claims, the singular forms encompass the plural forms unless otherwise clear from the context. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. [Brief explanation of the drawings]

[0064] [Figure 1] Figure 1 shows the effect of the BCAT2 inhibitor compound of formula Ia on BCAA pathway flux and on the downstream metabolite propionylcarnitine. The results show that in the presence of increasing concentrations of the BCAT2 inhibitor compound, propionylcarnitine levels are inhibited with little effect on cell viability. [Figure 2] Figure 2 shows plasma amino acid levels in mice treated with various doses of BCAT2 inhibitor compound (30, 100, and 300 mg / kg, PO) for 6 hours and then orally fed an amino acid mixture 1 hour before plasma collection. Each bar represents the mean ± SEM (N = 8-9 per group). *P < 0.05, **P < 0.01, ***P < 0.001 compared to vehicle control. In each set of histograms, the leftmost bar represents plasma levels for vehicle; the second bar represents plasma levels for 30 mg / kg of BCAT2 inhibitor compound; the third bar represents plasma levels for 100 mg / kg of BCAT2 inhibitor compound; and the rightmost bar represents plasma levels for 300 mg / kg of BCAT2 inhibitor compound. In Figure 2, the conditions are ordered (left to right): vehicle, 30 mg / kg BCAT, 100 mg / kg BCAT, and 300 mg / kg BCAT. [Figure 3-1] Figures 3A-D show (A) monitoring of body weight during 9 days of BCAT2 inhibitor compound treatment (200 mg / kg, PO, once daily) and a 3-day recovery period. Mice had free access to food and water throughout the study. Five days after the first dose, half of the animals were switched to a precursor-rich diet. The other half remained on a normal diet. (B) Body weights measured on day 0, (C) day 9, and (D) day 12 are shown. Each bar represents the mean ± SEM. Points represent individual values. In each set of histograms in Figures 3B-D, the left bar represents vehicle treatment, and the right bar represents BCAT2 inhibitor compound treatment. [Figure 3-2] See description of Figure 3-1. [Figure 4]Figures 4A-B show mouse plasma levels of valine, isoleucine, leucine, and serine on day 9. Mice were treated with vehicle or a BCAT2 inhibitor compound (200 mg / kg, PO, once daily) for 9 days and had free access to food and water throughout the study. Five days after the first dose, half of the animals were switched to a precursor-enriched diet (B). The other half remained on a normal diet (A). Each bar represents the mean ± SEM. Points represent individual values. N=8 / group. *P<0.05, **P<0.01 compared to vehicle control. For each set of histograms, the left bar represents vehicle dosing, and the right bar represents BCAT2 inhibitor compound dosing. [Figure 5] Figures 5A-B show plasma (5A) and urinary (5B) amino acid levels in day 9 samples from mice fed either a normal diet or a precursor-rich diet. Figures 5A-B show that BCAA overload by feeding a precursor-rich diet leads to increased renal excretion of these amino acids. In each set of histograms, the left bar represents the normal diet and the right bar represents the precursor-rich diet. DETAILED DESCRIPTION OF THE INVENTION

[0065] Detailed Description In some aspects, provided herein are compounds of Formula (I), (Ia), (II), (III), (IV), (V), (VI), (VII), and additional compounds. The compounds disclosed herein are BCAT2 inhibitors.

[0066] The present disclosure is based, in part, on the discovery that inhibition of targets within the branched-chain amino acid (BCAA) metabolic pathway may be used to identify inhibitors that will successfully reduce the substrate toxicity burden in patients suffering from organic acidemia. The present disclosure further provides methods for treating organic acidemia by reducing the levels of toxic metabolites and / or substrate molecules by administering a BCAT2 inhibitor compound disclosed herein to a patient in need thereof. The present disclosure also provides, in part, methods for treating organic acidemia by administering a BCAT2 inhibitor and an inhibitor of an amino acid transporter, including an SLC6A19 inhibitor, to a patient in need thereof.

[0067] In some embodiments of the present disclosure, patients receiving a BCAT inhibitor compound (e.g., a BCAT2 inhibitor compound) also receive an inhibitor of an amino acid transporter, including an inhibitor of SLC6A19 (e.g., the amino acid transporter B0 AT1), to treat and / or prevent hyperaminoacidemia. In various embodiments, administration of an inhibitor of an amino acid transporter, including an inhibitor of SLC6A19, reduces the amino acid load of patients receiving treatment with a BCAT2 inhibitor compound.

[0068] In one aspect, the present disclosure provides a method for identifying whether a candidate compound, including a candidate compound for treating an organic acidemia, is useful for treating an organic acidemia, comprising: (a) obtaining a test compound capable of binding directly or indirectly to BCAT (e.g., BCAT2); (b) assaying for functional modulation of BCAT (e.g., BCAT2); and (c) classifying the test compound as a candidate compound if decreased BCAT (e.g., BCAT2) activity is detected, low BCAT2 activity is detected, or substantially no BCAT (e.g., BCAT2) activity is detected.

[0069] In another aspect, the present disclosure provides a method for producing a medicament for treating organic acidemia, comprising: (a) identifying a candidate compound, comprising: (i) obtaining a test compound capable of binding directly or indirectly to BCAT (e.g., BCAT2); (ii) assaying for functional modulation of BCAT (e.g., BCAT2); and (iii) classifying the test compound as a candidate compound if a decrease in BCAT (e.g., BCAT2) activity is detected, low BCAT2 activity is detected, or substantially no BCAT (e.g., BCAT2) activity is detected; and (b) formulating the candidate compound for treating organic acidemia.

[0070] In another aspect, the present disclosure provides a method for treating or preventing organic acidemia, comprising administering to a subject in need thereof an effective amount of a compound disclosed herein.

[0071] In another aspect, the disclosure provides a method for treating or preventing organic acidemia, comprising administering to a subject in need thereof an effective amount of a compound selected from Table 1.

[0072] In another aspect, the present disclosure provides a method for reducing toxic branched chain amino acid (BCAA) metabolites, comprising administering to a subject in need thereof an effective amount of a compound disclosed herein.

[0073] In another aspect, the present disclosure provides a method for reducing toxic branched-chain amino acid (BCAA) metabolites, comprising administering to a subject in need thereof an effective amount of a compound selected from Table 1.

[0074] In another aspect, the present disclosure provides a method of reducing intracellular toxic load in a patient with organic acidemia, comprising administering to a subject in need thereof an effective amount of a compound disclosed herein.

[0075] In another aspect, the disclosure provides a method of reducing intracellular toxic burden in a patient with organic acidemia, comprising administering to a subject in need thereof an effective amount of a compound selected from Table 1.

[0076] [Table 1] TIFF0007789565000149.tif202167TIFF0007789565000150.tif195167TIFF0007789565000151.tif215167TIFF00077895650 00152.tif215167TIFF0007789565000153.tif199167TIFF0007789565000154.tif216167TIFF0007789565000155.tif249167

[0077] In another aspect of the present disclosure, the compound can be selected from the group consisting of compounds 106-306 as shown above and pharmaceutically acceptable salts thereof.

[0078] In some embodiments, the methods of the present disclosure further comprise administering an additional therapeutic agent, hi various embodiments, the additional therapeutic agent is an inhibitor of an amino acid transporter, including an inhibitor of SLC6A19.

[0079] In another aspect, the disclosure provides the use of a compound of Formula (I), (II), (III), (IV), (V), (VI), (VII) or any compound disclosed herein or a pharmaceutically acceptable salt thereof, alone or in combination with an additional therapeutic agent, in the manufacture of a medicament useful for the treatment or prevention of one or more organic acidemias.

[0080] Branched-chain amino acid metabolic pathways While not wishing to be bound by any one theory, it is currently believed that inhibiting the BCAT2 step of the BCAA pathway results in the systemic accumulation of branched-chain amino acids in the circulation, thereby preventing the formation of toxic metabolites that cannot be processed in patients with BCAA pathway mutations (e.g., patients with one or more organic acidemias). Therefore, it is postulated that substrate synthesis inhibition therapy results in a reduction in the toxic load within patient cells and has a transforming effect in alleviating their symptoms. Novel inhibitors could be developed as treatments for patients with rare organic acidemias under the FDA's Orphan Drug Program.

[0081] The branched-chain amino acids (BCAAs) are leucine, isoleucine, and valine. They share common membrane transport systems and enzymes for their transamination and irreversible oxidation. They can be glucogenic (valine), ketogenic (leucine and isoleucine), or both (isoleucine), as their end products, succinyl-CoA and / or acetyl-CoA, can enter the Krebs cycle for energy generation and gluconeogenesis or serve as precursors for lipogenesis and ketogenesis via acetyl-CoA and acetoacetate. Leucine, in particular, plays a central role in metabolism and participates in numerous signaling pathways. It is a potent stimulator of the mammalian target of rapamycin complex 1 and downstream targets that enhance translation elongation and protein synthesis. Leucine can also function as an inhibitor of muscle protein degradation through its interaction with the ubiquitin-proteasome and autophagy-lysosomal systems. Furthermore, leucine stimulates insulin secretion from pancreatic β-cells, functions as a metabolic fuel, and as an allosteric activator of glutamate dehydrogenase. Finally, it plays a role in the central nervous system's food intake regulation circuitry and feeding behavior.

[0082] The enzyme BCAT (e.g., mitochondrial BCAT isoenzyme, or "BCATm" or "BCAT2"; or cytosolic BCAT isoenzyme, or "BCATc" or "BCAT1") catalyzes the first step in the branched-chain amino acid catabolic pathway by a reversible transamination that is dependent on pyridoxal phosphate. This first step is followed by irreversible oxidative decarboxylation and coupled thioesterification of each keto acid by a single mitochondrial branched-chain α-keto acid dehydrogenase (BCKDH) complex to form a coenzyme A derivative. The oxidation of BCAAs and branched-chain α-keto acids (BCKAs) is tightly regulated, primarily at the BCKD step, which commits BCAAs to oxidative metabolism. The next step in the BCAA metabolic pathway is dehydrogenation of the activated keto acids by either isovaleryl-CoA dehydrogenase (leucine metabolism) or α-methyl-branched-chain dehydrogenase (isoleucine and valine metabolism). After these first three steps, the metabolism of each BCAA branches, ultimately yielding acetyl-CoA and / or propionyl-CoA. Terminal valine metabolism is unique because the free acid, 3-hydroxyisobutyric acid, is formed after hydrolysis of the corresponding thioester. 3-hydroxyisobutyric acid is dehydrogenated and then reacylated to complete the metabolism.

[0083] In various embodiments, the present disclosure contemplates a method for administering a BCAT2 inhibitor compound to a patient in need thereof, which causes the modulation of metabolic flux through the mitochondrial BCAA pathway (e.g., before or after treatment).In various embodiments, the present disclosure contemplates a method for administering a BCAT2 inhibitor compound to a patient in need thereof, which causes the reduction of metabolic flux through the mitochondrial BCAA pathway (e.g., before or after treatment).In various embodiments, the present disclosure contemplates a method for administering a BCAT2 inhibitor compound to a patient in need thereof, which causes the modulation of catabolic flux through the mitochondrial BCAA pathway (e.g., before or after treatment).More specifically, in some embodiments, it is contemplated that the method of the present disclosure inhibits and / or reduces BCAT2-mediated flux through the mitochondrial BCAA pathway.

[0084] For example, with respect to leucine, a reduction in BCAT2-mediated flux can occur when leucine metabolism is reduced and / or inhibited. In various embodiments, leucine metabolism is reduced and / or inhibited in one or more enzymes in the BCAA metabolic pathway selected from branched-chain α-keto acid dehydrogenase (BCKDH), isovaleryl-CoA dehydrogenase (IVD), 3-methylcrotonyl-CoA carboxylase (3MCC), 3-methylglutaconic-CoA hydratase (3MGA), 3-hydroxy-3-methylglutaric-CoA lyase (HMGL), acetyl-CoA carboxylase (AC), and malonyl-CoA decarboxylase (MA).

[0085] In a further embodiment, with respect to isoleucine, the reduction in BCAT2-mediated flux occurs when isoleucine metabolism is reduced or inhibited. In some embodiments, isoleucine metabolism is reduced and / or inhibited in one or more enzymes in the BCAA metabolic pathway selected from BCKDH, methylbutyryl-CoA dehydrogenase (SBCAD), hydratase, 2-methyl-3-hydroxyisobutyrate dehydrogenase (MHBD), acetoacetyl-CoA thiolase (T2), AC, MA, propionyl-CoA carboxylase (PCC), methylmalonyl-CoA mutase (MUT), and succinyl-CoA ligase (SUCLA).

[0086] In a further aspect, with respect to valine, the reduction in BCAT2-mediated flux occurs when valine metabolism is reduced and / or inhibited. In some aspects, valine metabolism is reduced and / or inhibited in one or more enzymes in the BCAA metabolic pathway selected from BCKDH, isobutyryl-CoA dehydrogenase (IBDH), hydratase, 3-hydroxyisobutyryl-CoA deacylase (hydrolase) (HIBDA), 3-hydroxyisobutyrate dehydrogenase (HIBDH), methylmalonate semialdehyde dehydrogenase (MMSDH), PCC, MUT, and SUCLA.

[0087] Methods for the treatment of organic acidemias In various aspects, the present disclosure provides a method for treating one or more organic acidemias.In such aspects, "agents of the present disclosure" include compounds that are useful in both monotherapy and combination therapy (for example, as additional therapeutic agents).Generally, monotherapy involves the use of a compound of formula I or its pharmaceutically acceptable salt, while combination therapy involves the use of a compound of formula I or its pharmaceutically acceptable salt in combination with additional therapeutic agents, such as amino acid transporter inhibitors, including SLC6A19 inhibitors.

[0088] In some embodiments, the present disclosure contemplates that administering an additional therapeutic agent, such as an inhibitor of an amino acid transporter, including an SLC6A19 inhibitor, reduces elevated branched-chain amino acid levels compared to treatment without administration of an inhibitor of an amino acid transporter, including an SLC6A19 inhibitor. In various embodiments, the administration of an inhibitor of an amino acid transporter, including an SLC6A19 inhibitor (e.g., separately or in combination with a BCAT2 inhibitor compound described herein), can reduce hyperaminoacidemia caused by BCAT2 inhibition by the methods of the present disclosure.

[0089] As used herein, organic acidemias are characterized as a group of inherited metabolic disorders that disrupt normal amino acid metabolism, particularly branched-chain amino acid metabolism, resulting in the accumulation of acids that are not normally present.

[0090] In some aspects, the organic acidemia is any form of methylmalonic acidemia (MMA), any form of propionic acidemia (PA), isovaleric acidemia, glutaric aciduria type 1 (GA1), beta-ketothiolase deficiency (BKT), 3-methylcrotonyl-CoA carboxylase deficiency (3-MCC), 3-hydroxy-3-methylglutaric-CoA lyase deficiency (HMG), 3-methylglutaconic acidemia, or 3-methylglutaconyl-CoA hydratase deficiency (M GA), D-2 hydroxyglutaric aciduria (D2-HGA), isobutyryl-CoA dehydrogenase deficiency 3-hydroxyisobutyric aciduria (ICBD), L-2-hydroxy-glutaric aciduria (L2HGA), malonyl-CoA decarboxylase deficiency, also known as malonic acidemia (MA), multiple carboxylase deficiency (MCD, holocarboxylase synthase) and 3-hydroxyisobutyryl-CoA hydrolase deficiency (HIBCH).

[0091] In various embodiments, the organic acidemia is methylmalonic acidemia or propionic acidemia.

[0092] Mutations in one or more components of the BCAA catabolic pathway can cause one or more organic acidemias.For example, various enzymes or components that function in the entire BCAA pathway can be mutated, resulting in the manifestation of organic acidemias.In some embodiments, the BCAA metabolic pathway of leucine includes one or more enzymes or components that can be mutated, such as branched-chain α-keto acid dehydrogenase (BCKDH), isovaleryl-CoA dehydrogenase (IVD), 3-methylcrotonyl-CoA carboxylase (3MCC), 3-methylglutaconic-CoA hydratase (3MGA), 3-hydroxy-3-methylglutaric-CoA lyase (HMGL), acetyl-CoA carboxylase (AC) and malonyl-CoA decarboxylase (MA). In some embodiments, the BCAA metabolic pathway of isoleucine includes enzymes or components that may be in a mutated state, such as one or more of BCKDH, methylbutyryl-CoA dehydrogenase (SBCAD), hydratase, 2-methyl-3-hydroxyisobutyrate dehydrogenase (MHBD), acetoacetyl-CoA thiolase (T2), AC, MA, propionyl-CoA carboxylase (PCC), methylmalonyl-CoA mutase (MUT), and succinyl-CoA ligase (SUCLA). In further embodiments, the BCAA metabolic pathway of valine includes enzymes or components that may be in a mutated state, such as one or more of BCKDH, isobutyryl-CoA dehydrogenase (IBDH), hydratase, 3-hydroxyisobutyryl-CoA deacylase (hydrolase) (HIBDA), 3-hydroxyisobutyrate dehydrogenase (HIBDH), methylmalonate semialdehyde dehydrogenase (MMSDH), PCC, MUT, and SUCLA.

[0093] Disorders of the branched-chain amino acid metabolic pathway are further described in Manoli and Venditti, Disorders of branched chain amino acid metabolism, Translational Science of Rare Diseases 1 (2016) 91-110 and Schiff et al., J.M. Saudubray et al. (Eds.), Branched-chain Organic Acidurias Inborn Metabolic Diseases, Springer Verlag Berlin (279-294) 2016, both of which are incorporated herein by reference in their entireties.

[0094] In some embodiments, the organic acidemia is selected from the group consisting of maple syrup urine disease type Ia; maple syrup urine disease type Ib; maple syrup urine disease type II; maple syrup urine disease: mild variant; isovaleric acidemia; 3-methylcrotonyl-CoA carboxylase deficiency; 3-methylcrotonyl-CoA carboxylase 2 deficiency; 3-methylglutaconic aciduria type I; 3-methylglutaconic aciduria type II: Barth syndrome; 3-methylglutaconic aciduria type III: Costeff syndrome; 3-methylglutaconic aciduria type IV: "unclassifiable type"; 3-methylglutaconic aciduria type V; 3-methylglutaconic aciduria type VI: hearing loss, encephalopathy, Leigh-like syndrome; 3-methylglutaconic aciduria type VII: cataracts, neuroinfiltration and neutropenia; 3-hydroxybenzoates The deficiency is selected from one or more of: 2-methyl-3-hydroxybutyryl-CoA dehydrogenase deficiency; mitochondrial short-chain enoyl-CoA hydratase-1 deficiency; methylbutyryl-CoA dehydrogenase deficiency; mitochondrial acetoacetyl-CoA thiolase deficiency; isobutyryl-CoA dehydrogenase deficiency; 3-hydroxyisobutyryl-CoA deacylase deficiency; 3-hydroxyisobutyric aciduria; methylmalonic semialdehyde dehydrogenase deficiency; propionic acidemia, methylmalonic acidemia: Mut subtype; methylmalonic acidemia: cobalamin A; methylmalonic acidemia: cobalamin B; and methylmalonic acidemia: cobalamin D.

[0095] In various embodiments, the method of the present disclosure is assumed to reduce the level of toxic substrate metabolites that occur in the BCAA metabolic pathway.Toxic substrate metabolites can include but are not limited to propionic acid, 2-methylcitric acid, 3-hydroxypropionic acid, propionyl-CoA, methylmalonic acid, methylmalonyl-CoA and isovaleric acid.

[0096] Methylmalonic acidemia Methylmalonic acidemia ("MMA") is an autosomal recessive disorder caused by a defect in the mitochondrial enzyme methylmalonyl-CoA mutase (MUT). The estimated incidence of MMA is 1 in 25,000 to 48,000 individuals. MUT is an enzyme that catalyzes the conversion of L-methylmalonyl-CoA to succinyl-CoA. This reaction is one of several required to metabolize branched-chain amino acids, odd-chain fatty acids, cholesterol, and propionate produced by intestinal flora. MUT deficiency, the most common cause of isolated MMA, is characterized by the accumulation of methylmalonic acid and other toxic metabolites. MMA is associated with metabolic instability, seizures, pancreatitis, stroke, and renal failure, and can be fatal even when patients are appropriately managed, highlighting the need for new therapeutic agents for this disease. Current treatments for MMA include, but are not limited to, dietary restriction, liver transplantation, combined liver and kidney transplantation, and liver-directed gene therapy.

[0097] The MUT enzyme requires adenosylcobalamin (Ado-Cbl) as a coenzyme. Therefore, methylmalonate metabolism is necessarily linked to vitamin B12 (cobalamin), its adequate intake, proper uptake, transport, and intracellular metabolism. Complementation of the variant 2 forms of cblA, cblB, and cblD is associated with a unique process in Ado-Cbl synthesis. Complementation of the variants cblC, cblD, cblF, and cblJ is similarly associated with defective methyl-cobalamin synthesis. Cobalamin C (cblC), a combined form of MMA and homocystinuria, is the most common disorder of cobalamin metabolism. CblC typically manifests in the neonatal period and is associated with neurological decline, growth failure, cytopenias, and multiorgan pathology, including renal and hepatic dysfunction.

[0098] Propionic acidemia A related disorder, propionic acidemia ("PA"), is an autosomal recessive disorder caused by deficiencies in either propionyl-CoA carboxylase ("PCC"), either the alpha (PCCA) or beta subunit (PCCB). PCC is inactive in affected individuals with either PCCA or PCCB deficiency. Patients with PA are unable to metabolize branched-chain amino acids, odd-chain fatty acids, cholesterol, and propionate, which are produced by the intestinal flora. This condition leads to the abnormal accumulation of certain acids, known as organic acids, including propionic acid, 2-methylcitric acid, and 3-hydroxypropionic acid. As a result, a substance called propionyl-CoA and other potentially harmful compounds can build up in the body to toxic levels. This accumulation damages the brain, nervous system, and heart, leading to the serious health problems associated with PA. PA is associated with metabolic instability, seizures, pancreatitis, stroke, and a tendency toward hyperammonemia. Treatments for PA include, but are not limited to, dietary restriction, liver transplantation, and liver-directed gene therapy. Like MMA, PA can be fatal even when patients are properly managed.

[0099] Maple syrup urine disease Maple syrup urine disease ("MSUD") is a rare genetic disorder characterized by a deficiency of certain enzymes (e.g., branched-chain alpha-ketoacid dehydrogenase complex) required to break down (metabolize) three branched-chain amino acids. In the classic, severe form of MSUD, plasma concentrations of BCAAs begin to increase within hours of birth. Without treatment, symptoms often begin within 24–48 hours of birth. Classic MSUD is associated with lethargy and / or irritability, progressive encephalopathy, opisthotonos, and coma. Intermediate MSUD is associated with stress-induced metabolic encephalopathy, anorexia, and growth failure. Intermittent MSUD is associated with normal early development and recurrent crises associated with stress. Type III E3-deficient MSUD is associated with Leigh-type encephalopathy, lactic acidosis, and often death.

[0100] Compounds of the Disclosure In various aspects, the present disclosure provides for the identification and use of candidate compounds. In embodiments providing for the identification and use of candidate compounds, the candidate compounds can be chemicals, molecules, compounds, biological agents (e.g., antibodies or peptides), drugs, prodrugs, cell therapy drugs, low molecular weight synthetic compounds, or small molecule drugs. In some embodiments, the candidate compounds are selected from libraries of compounds known in the art. In some embodiments, the candidate compounds are useful for treating organic acidemia and / or preventing organic acidemia.

[0101] In some aspects, there is provided herein a compound of formula (I) or a pharmaceutically acceptable salt thereof: TIFF0007789565000156.tif42128In formula, Z can be CH2, CF2, O or a bond; J 1 , J 2 , J 3 and J. 4 each may independently be CH or N; X 1 is N or CR 5A It can be; X 2 is N or CR 5B It can be; R 1 is C1-C6 alkyl, -NR 3 S(O)R 4 , -NR 3 SO2R 4 , -NR 3 C(O)R 4 , -C(O)(CH2) m NR 3 R 4 , C6~C 10 may be aryl, 5-10 membered heteroaryl or (5-10 membered heteroaryl)-C1-C6 alkyl; R 2 is -NR 6 R 7 , C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10It can be an aryl or a 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO2, C1-C6 alkyl, C1-C6 alkoxy, haloC1-C6 alkyl, haloC1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC1-C6 alkyl), C6-C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' the substituents, together with the atom to which they are attached, form a 5-membered heterocyclyl or C3-C8 cycloalkyl, each optionally substituted with halo; R 3 can be H or C1-C6 alkyl; R 4 is H, C1-C6 alkyl, C2-C6 alkenyl, C6-C 10 It can be an aryl or a 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, C6-C 10 Aryl, -(CH2) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with; R 5A is H, halo, -CN, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, -(CH2) r OH, -C(O)NR8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 It can be; R 5B is H, halo, -CN, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, -(CH2) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 Could it be; Or R 5A and R 5B may be taken together with the atom to which they are attached to form a 6-membered heterocyclyl ring optionally substituted with oxo; R 6 and R 7 can independently be H, C1-C6 alkyl, or C3-C8 cycloalkyl; Each R 8 may independently be H, C1-C6 alkyl, or C3-C8 cycloalkyl; Each R 9 can independently be H, C1-C6 alkyl, C2-C6 alkenyl, or C3-C8 cycloalkyl; m can be 0 or 1; q can be 0 or 1; r can be 0 or 1 The present invention provides a compound or a pharmaceutically acceptable salt thereof.

[0102] In some embodiments, R 5A is not hydrogen.

[0103] In some embodiments, R 5B is hydrogen or -C(O)NR 8 R 9 No.

[0104] In some embodiments, R 2 is not unsubstituted aryl or unsubstituted heteroaryl. 2 is not aryl substituted with one -S-C1-C6 alkyl. 2 is not a 5-10 membered heteroaryl substituted with one C1-C6 alkyl.

[0105] In some embodiments, the compound of formula (I) is TIFF0007789565000157.tif211147TIFF0007789565000158.tif200147 and pharmaceutically acceptable salts and stereoisomers thereof is not selected from the group consisting of:

[0106] In some embodiments, Z can be CH. In other embodiments, Z can be CF. In still other embodiments, Z can be O. In still other embodiments, Z can be a bond.

[0107] In some embodiments, J 1 can be CH. In other embodiments, J 1 can be N.

[0108] In some embodiments, J 2 can be CH. In other embodiments, J 2 can be N.

[0109] In some embodiments, J 3 is CH. In other embodiments, J 3 can be N.

[0110] In some embodiments, J 4 can be CH. In other embodiments, J 4 can be N.

[0111] In some embodiments, X 1 can be N. In other embodiments, X 1 is CR 5A It could be.

[0112] In some embodiments, X 2 can be N. In other embodiments, X 2 is CR 5B It could be.

[0113] In some embodiments, J 2 can be N, and X 1 can be N. In other embodiments, J 2 can be N, and J 3 can be N. In other embodiments, J 2 can be N, and J 4 can be N. In still other embodiments, J 2 can be N, and X 2 can be N. In some embodiments, J 2 , J 3 and J. 4 Each of X may be CH; 1 is CR 5A X 2 is CR 5B It could be.

[0114] In some embodiments, R 1 NR 3 C(O)R 4 In other embodiments, R 1 -C(O)(CH2) m NR 3 R 4 It could be.

[0115] In some embodiments, R 3 can be hydrogen. In other embodiments, R 3 C 1~6 It can be alkyl.

[0116] In some embodiments, R 4 each containing one, two or three substituents R 4' C optionally substituted with 6~10 aryl or 5-10 membered heteroaryl. In some embodiments, R 4may be selected from phenyl, thiophene, oxazole, isoxazole, thiazole, furan and pyrrole, each of which may contain one, two or three substituents R 4' where each R 4' may be independently selected from the group consisting of -F, -Cl, -Br, -CN, -NO2, -CF3, -CH3, -CH(CH3)2, -C(O)CH3, -C(O)OCH3, phenyl, cyclopropyl, and morpholinyl.

[0117] In some embodiments, m can be 0. In other embodiments, m can be 1.

[0118] In some embodiments, R 2 each containing one, two or three substituents R 2' C optionally substituted with 6~10 aryl or 5-10 membered heteroaryl. In some embodiments, R 2 may be selected from the group consisting of phenyl, pyridine, pyrimidine, pyridazine, pyrazine, imidazole or thiophene, each of which may contain one, two or three substituents R 2' where each R 2' may be independently selected from the group consisting of -F, -Cl, -Br, -CN, NO, -CH, -CFH, -C≡CH, -C(O)H, -CONH, -C(O)NHCH, -OH, -OCH, -OCF, -SCF, -NH, -NHC(O)CH, and morpholine.

[0119] In some embodiments, X 2 is CR 5B R 5B -C(O)NR 8 R 9 In other embodiments, X 2 is CR 5B R 5B halo, -CN, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, -(CH2) r OH, -NR 8 COR 9, 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 In yet another embodiment, X 2 is CR 5B R 5B Ga-NR 8 COR 9 In some embodiments, X 2 is CR 5B R 5B -C(O)OR 8 In some embodiments, X 2 is CR 5B R 5B can be -CF3, -COCH3, -CH2OH, -CN or tetrazole.

[0120] In some embodiments, R 8 can be hydrogen. In other embodiments, R 8 Ga-C 1~6 It can be alkyl.

[0121] In some embodiments, R 9 can be hydrogen. In other embodiments, R 9 Ga-C 1~6 It can be alkyl.

[0122] In some embodiments, compounds of formula (Ia) are described herein: a compound of formula (I) having the structure: TIFF0007789565000159.tif34128, or a pharmaceutically acceptable salt thereof; During the ceremony, R 1 is C1-C6 alkyl, -NR 3 S(O)R 4 , -NR 3 SO2R 4 , -NR 3 C(O)R 4 , -C(O)(CH2) m NR 3 R 4 , C6~C 10 may be aryl or 5-10 membered heteroaryl; R2 is C1-C6 alkyl, C6-C 10 may be aryl or 5-10 membered heteroaryl; R 4 is C6~C 10 may be aryl or 5-10 membered heteroaryl; R 5B is -C(O)NR 8 R 9 It can be; R 8 and R 9 are independently H, C1-C6 alkyl, or C3-C8 cycloalkyl.

[0123] In some aspects, the term "aromatic group" is used herein to refer to a group comprising a compound of formula: R 1 Ga-NR 3 SO2R 4 or -NR 3 C(O)R 4 It can be; R 2 can be aryl or 5-10 membered heteroaryl; R 3 can be H or C1-C6 alkyl; R 4 C6~C 10 may be aryl or 5-10 membered heteroaryl; R 5B -C(O)NR 8 R 9 It can be; R 8 and R 9 can independently be H or C1-C6 alkyl Provided is a compound of formula (Ia) or a pharmaceutically acceptable salt thereof:

[0124] In some aspects, the term "aromatic group" is used herein to refer to a group comprising a compound of formula: R 1 is -NR3C(O)R4; R 2 C6~C 10 aryl or 5-10 membered heteroaryl; R 3 is H or methyl; R 4 C6~C 10 aryl or 5-10 membered heteroaryl; R 5B -C(O)NR 8 R 9 and; R 8 and R 9 are independently H or C1-C6 alkyl Provided is a compound of formula (Ia) or a pharmaceutically acceptable salt thereof:

[0125] In some aspects, the term "aromatic group" is used herein to refer to a group comprising a compound of formula: R1 is -NR3C(O)R4; R 2 C6~C 10 aryl or 5-10 membered heteroaryl; R 3 is H or methyl; R 4 C6~C 10 aryl or 5-10 membered heteroaryl; R 5 -C(O)NR 8 R 9 and; R 8 and R 9 are independently H or methyl Provided is a compound of formula (Ia) or a pharmaceutically acceptable salt thereof:

[0126] In some aspects, the compounds of the present disclosure are 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(2-(methylthio)phenyl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-phenyl-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-3-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-4-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiophen-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiophen-3-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(2-methylthiazol-4-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiazol-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(1H-pyrazol-3-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-cyanothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-chlorothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-methylthiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromo-N-methylthiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-((5-bromothiophene)-2-sulfonamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 2-bromo-N-(3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)thiazole-5-carboxamide, 1-(3-(5-bromo-1-methyl-1H-pyrrole-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(4-bromo-1H-pyrrole-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 3-chloro-N-(3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)isoxazole-5-carboxamide, 5-bromo-N-(3-(2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)thiophene-2-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N,N-dimethyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-cyclopropyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide, and 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-isopropyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide; or a pharmaceutically acceptable salt thereof It could be.

[0127] In a further aspect, there is provided herein a compound of formula (II): There may be provided a compound of formula (I) having the structure of TIFF0007789565000160.tif48128, or a pharmaceutically acceptable salt thereof, wherein: R 2 is C6~C 10 It can be an aryl or a 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO2, C1-C6 alkyl, C1-C6 alkoxy, haloC1-C6 alkyl, haloC1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC1-C6 alkyl), C6-C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' the substituents, together with the atom to which they are attached, may form a 5-membered heterocyclyl or a C3-C8 cycloalkyl, each optionally substituted with halo; R 4 is halo, -CN, -NO2, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, C6-C 10Aryl, -(CH2) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be a 5-10 membered heteroaryl optionally substituted with R 5B is -C(O)NR 8 R 9 It could be.

[0128] In some embodiments, R 2 is one, two, or three substituents R independently selected from halo, —CN, —NO2, and C1-C6 alkyl; 2' In other embodiments, R 2 is one, two, or three substituents R independently selected from halo, —CN, —NO2, and C1-C6 alkyl; 2' and 5-10 membered heteroaryl optionally substituted with

[0129] In some embodiments, R 4 may be a thiophene optionally substituted with halo or phenyl.

[0130] In a further aspect, there is provided herein a compound of formula (III): There may be provided a compound of formula (I) having the structure: TIFF0007789565000161.tif50128, or a pharmaceutically acceptable salt thereof: During the ceremony, each J 3 and J. 4 are independently CH or N; X 1 is N or CR 5A and; X 2 is N or CR 5B and; Here, J 3 , J4 , X 1 and X 2 of which 1, 2, 3 or 4 are N; R 2 is C6~C 10 aryl or 5-10 membered heteroaryl, wherein each aryl or heteroaryl is halo, -CN, -NO2, C1-C6 alkyl, C1-C6 alkoxy, haloC1-C6 alkyl, haloC1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S-(haloC1-C6 alkyl), C6-C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' the substituents, together with the atom to which they are attached, form a 5-membered heterocyclyl or C3-C8 cycloalkyl, each optionally substituted with halo; R 4 is H, C1-C6 alkyl, C2-C6 alkenyl, C6-C 10 aryl or 5-10 membered heteroaryl, wherein each aryl or heteroaryl is selected from halo, -CN, -NO2, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, C6-C 10 Aryl, -(CH2) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two, or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with.

[0131] In some embodiments, R 2 is one, two, or three substituents R independently selected from halo, —CN, —NO2, and C1-C6 alkyl; 2' In other embodiments, R 2 is one, two, or three substituents R independently selected from halo, —CN, —NO2, and C1-C6 alkyl; 2' and 5-10 membered heteroaryl optionally substituted with

[0132] In some embodiments, R 4 may be a thiophene optionally substituted with halo or phenyl.

[0133] In some embodiments, X 2 is CR 5B R 5B -H, -halo, -C(O)NR 8 R 9 , -NR 8 COR 9 or -C(O)OR 8 R 8 and R 9 may each independently be H or C1-C6 alkyl.

[0134] In a further aspect, there is provided herein a compound of formula (IV): There may be provided a compound of formula (I) having the structure of TIFF0007789565000162.tif44128, or a pharmaceutically acceptable salt thereof, wherein: R 1 is C1-C6 alkyl, -NR 3 C(O)R 4 , -C(O)(CH2) m NR 3 R 4 , C6~C 10 aryl, 5-10 membered heteroaryl, or (5-10 membered heteroaryl)-C1-C6 alkyl; R 2 Ha-NR 6 R 7 , C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl or 5-10 membered heteroaryl, wherein each aryl or heteroaryl is halo, -CN, -NO2, C1-C6 alkoxy, haloC1-C6 alkyl, haloC1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, C6~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, form a 5-membered heterocyclyl or C3-C8 cycloalkyl, each optionally substituted with halo.

[0135] In still a further aspect, there is provided herein a compound of formula (V): There may be provided a compound of formula (I) having the structure: TIFF0007789565000163.tif44128, or a pharmaceutically acceptable salt thereof: During the ceremony, R 5A is halo, -CN, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, -(CH2) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 and; R 5B is H, halo, -CN, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, -(CH2) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8or -C(O)OR 8 Is it; Or R 5A and R 5B are taken together with the atoms to which they are attached to form a 6-membered heterocyclyl ring which may be substituted with oxo.

[0136] In still yet a further aspect, there is provided herein a compound of formula (VI): There may be provided a compound of formula (I) having the structure TIFF0007789565000164.tif45128, or a pharmaceutically acceptable salt thereof.

[0137] In a still yet further aspect, there is provided herein a compound of formula (VII): There may be provided a compound of formula (I) having the structure TIFF0007789565000165.tif46128, or a pharmaceutically acceptable salt thereof.

[0138] In a still yet further aspect, there is provided herein a compound selected from the group consisting of compounds 106-306, as described above, and pharmaceutically acceptable salts thereof.

[0139] The compound of the present disclosure, which may be in the form of free base, can be isolated from the reaction mixture in the form of pharmaceutically acceptable salt.Also, pharmaceutically acceptable salt can be obtained by treating the compound of the present disclosure in the form of free base with organic acid or inorganic acid, such as hydrogen chloride, hydrogen bromide, hydrogen iodide, sulfuric acid, phosphoric acid, acetic acid, propionic acid, glycolic acid, maleic acid, malonic acid, methanesulfonic acid, fumaric acid, succinic acid, tartaric acid, citric acid, benzoic acid and ascorbic acid.

[0140] When a compound disclosed herein contains at least one chiral center, it may exist as individual enantiomers and diastereomers, or as a mixture of such isomers, e.g., a racemate. Separation of individual isomers or selective synthesis of individual isomers can be achieved by applying various methods known to those skilled in the art. Unless otherwise indicated, all such isomers and mixtures thereof are included within the scope of the compounds disclosed herein. Furthermore, compounds disclosed herein may exist in one or more crystalline or amorphous forms. Unless otherwise indicated, all such forms, including any polymorphic forms, are included within the scope of the compounds disclosed herein. Additionally, some compounds disclosed herein may form solvates with water (i.e., hydrates) or common organic solvents. Unless otherwise indicated, such solvates are included within the scope of the compounds disclosed herein.

[0141] Those skilled in the art will recognize that some structures depicted herein may be resonance forms or tautomers of compounds that may be appropriately represented, even if kinetically, by other chemical structures; those skilled in the art will recognize that such structures may represent only a very small portion of a sample of such compound(s). Such compounds are considered to be within the scope of the depicted structures, even though such resonance forms or tautomers are not depicted herein.

[0142] Isotopes may be present in the compounds described herein. Each chemical element shown in the compound structure may include any isotope of said element. Isotopes include isotopes of carbon, chlorine, fluorine, hydrogen, iodine, nitrogen, oxygen, phosphorus, sulfur, and technetium, e.g., 11 C. 13 C. 14 C. 36 Cl, 18 F, 2 H, 3 H, 123 I, 125 I, 13 N, 15 N, 15 O. 17 O.18 O. 31 P, 32 P, 35 S and 99m Tc. For example, in the structure of a compound, hydrogen atoms may be explicitly disclosed or may be understood to be present in the compound. At any position where a hydrogen atom of the compound may be present, the hydrogen atom may be any isotope of hydrogen, for example, but not limited to, hydrogen-1 (protium) and hydrogen-2 (deuterium). Therefore, reference to a compound herein encompasses all possible isotopic forms, unless the context makes clear otherwise.

[0143] chemical synthesis method The compounds encompassed by the present disclosure can be prepared by methods well known in the art of organic chemistry. See, for example, March, Advanced Organic Chemistry, 4th Edition, John Wiley & Sons, 2001. During the synthetic sequence, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules involved. This is accomplished by conventional protecting groups, such as those described in Greene and Wutts, Protective Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons, 1999. The protecting groups are optionally removed at a subsequent convenient stage using methods well known in the art.

[0144] The products of the reactions are optionally isolated and purified, if desired, using conventional techniques, including, but not limited to, filtration, distillation, crystallization, chromatography, etc. Such materials are optionally characterized using conventional means, for example, measurement of physical constants and spectral data.

[0145] The synthesis of the compounds is represented by the following general scheme: Detailed synthesis methods can be found in the Examples section. TIFF0007789565000166.tif106128

[0146] Further synthesis of the compounds is represented by the following general scheme: Detailed synthesis methods can be found in the Examples section. TIFF0007789565000167.tif110154

[0147] Method for identifying candidate compounds The present disclosure provides a method for identifying whether a candidate compound, including a candidate compound for treating an organic acidemia, is useful for treating an organic acidemia, comprising, in part: (a) obtaining a test compound capable of binding directly or indirectly to BCAT (e.g., BCAT2); (b) assaying for functional modulation of BCAT (e.g., BCAT2); and (c) classifying the test compound as a candidate compound if decreased BCAT (e.g., BCAT2) activity is detected, low BCAT2 activity is detected, or substantially no BCAT (e.g., BCAT2) activity is detected.

[0148] In various embodiments, the present disclosure envisions the use of various assays to classify candidate compounds for use in the functional modulation of BCAT2.For example, the functional modulation of BCAT2 can be measured using cell-based assays.In some embodiments, when the increase of branched-chain amino acids is measured, the test compound is classified as candidate compound.In some embodiments, when the decrease of propionyl-carnitine is measured, the test compound is classified as candidate compound.

[0149] In further embodiments, the functional modulation of BCAT2 is assayed using a cell-free assay. In various embodiments, the cell-free assay is an enzyme-coupled fluorescent assay. For example, the present disclosure contemplates the use of an assay that monitors the production of L-glutamate from branched-chain amino acids and α-ketoglutarate by coupling hBCATm activity with two additional enzymes, L-glutamate oxidase (L-GOx) and horseradish peroxidase (HRP). Specifically, L-GOx catabolizes L-glutamate to produce α-ketoglutarate and HRP, which is utilized by HRP to form fluorescent resorufin via the redox-sensitive dye Amplex Red. In some embodiments, if a decrease in L-glutamate is measured, the test compound is classified as a candidate compound. In further embodiments, if a decrease in α-ketoglutarate is measured, the test compound is classified as a candidate compound.

[0150] Methods for producing the medicament of the present disclosure The present disclosure provides a method for producing a medicament for treating organic acidemia, comprising: (a) identifying a candidate compound, comprising: (i) obtaining a test compound capable of binding directly or indirectly to BCAT (e.g., BCAT2); (ii) assaying for functional modulation of BCAT (e.g., BCAT2); and (iii) classifying the test compound as a candidate compound if a decrease in BCAT (e.g., BCAT2) activity is detected, low BCAT2 activity is detected, or substantially no BCAT (e.g., BCAT2) activity is detected; and (b) formulating the candidate compound for treating organic acidemia.

[0151] Pharmaceutically acceptable salts and excipients Any of the drugs described herein may have a sufficiently basic functional group that can react with inorganic or organic acids, or a carboxyl group that can react with inorganic or organic bases to form pharmaceutically acceptable salts. Pharmaceutically acceptable acid addition salts are formed with pharmaceutically acceptable acids, as is well known in the art. Such salts include those listed in Journal of Pharmaceutical Science, 66, 2-19 (1977) and The Handbook of Pharmaceutical Salts: Properties, Selection, and Use. P.H. Stahl and C.G. Wermuth (eds.), Verlag, Zurich (Switzerland), 2002, the entire contents of which are incorporated herein by reference.

[0152] Pharmaceutically acceptable salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, isopropyl betaine, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharinate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, pamoate, phenylacetate, trifluoroacetate, acrylate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetate, benzoic acid ... Examples of suitable salts include benzoxybenzoate, naphthalene-2-benzoate, isobutyrate, phenylbutyrate, alpha-hydroxybutyrate, butyne-1,4-dicarboxylate, hexyne-1,4-dicarboxylate, caprate, caprylate, cinnamate, glycolate, heptanoate, hippurate, malate, hydroxymaleate, malonate, mandelate, mesylate, nicotinate, phthalate, teraphthalate, propiolate, propionate, phenylpropionate, sebacate, suberate, p-bromobenzenesulfonate, chlorobenzenesulfonate, ethylsulfonate, 2-hydroxyethylsulfonate, methylsulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, naphthalene-1,5-sulfonate, xylenesulfonate, and tartrate.

[0153] The term "pharmaceutically acceptable salt" also refers to a salt of a compound of the present disclosure having an acidic functional group, such as a carboxylic acid functional group, with a base. Suitable bases include, but are not limited to, hydroxides of alkali metals such as sodium, potassium, and lithium; hydroxides of alkaline earth metals such as calcium and magnesium; hydroxides of other metals such as aluminum and zinc; ammonia, and organic amines such as unsubstituted or hydroxy-substituted mono-, di-, or tri-alkylamines, dicyclohexylamine; tributylamine; pyridine; N-methyl, N-ethylamine; diethylamine; triethylamine; mono-, bis-, or tris-(2-OH-lower alkylamines), such as mono-, bis-, or tris-(2-hydroxyethyl)amine, 2-hydroxy-tert-butylamine, or tris-(hydroxymethyl)methylamine, N,N-di-lower alkyl-N-(hydroxyl-lower alkyl)-amines, such as N,N-dimethyl-N-(2-hydroxyethyl)amine or tri-(2-hydroxyethyl)amine; N-methyl-D-glucamine; and amino acids such as arginine, lysine, and the like.

[0154] In some embodiments, the agents of the present disclosure are in the form of or a pharmaceutically acceptable salt. In some embodiments, the pharmaceutically acceptable salt is a sodium salt.

[0155] Additionally, any agent described herein can be administered to a subject as a component of a composition comprising a pharmaceutically acceptable carrier or vehicle. Such compositions can optionally contain a suitable amount of a pharmaceutically acceptable excipient so as to provide the form for proper administration.

[0156] Pharmaceutical excipients can be liquids, such as water and oils, such as those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Pharmaceutical excipients can be saline, acacia gum, gelatin, starch paste, talc, keratin, colloidal silica, urea, etc. Auxiliaries, stabilizers, thickeners, lubricants, and coloring agents can also be used. In one embodiment, the pharmaceutically acceptable excipients are sterile when administered to a subject. Water is a useful excipient when any of the agents described herein are administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be used as liquid excipients, particularly for injectable solutions. Suitable pharmaceutical excipients also include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, nonfat dry milk, glycerol, propylene, glycol, water, ethanol, etc. Any of the agents described herein, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents.

[0157] Formulation, Administration and Dosing Any of the agents described herein may be administered orally. They may also be administered by any other convenient route, such as by intravenous infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.), or may be administered together with another bioactive agent. Administration may be systemic or local. Various delivery systems, such as encapsulation in liposomes, microparticles, microcapsules, capsules, etc., are known and may be used for administration.

[0158] In certain embodiments, it may be desirable to administer locally to the area where treatment is required.

[0159] In another embodiment, delivery can be in vesicles, particularly liposomes (see Langer, 1990, Science 249:1527-1533; Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989)). In yet another embodiment, delivery can be in a controlled release system. In one embodiment, a slow release intraocular device can be used. In some embodiments, the device is comprised of a locally delivered disintegrating or non-disintegrating liquid, gel, polymer, or the like.

[0160] In another embodiment, polymeric materials can be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Florida (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger and Peppas, 1983, J. Macromol. Sci. Rev. Macromol. Chem. 23:61; see also Levy et al., 1985, Science 228:190; During et al., 1989, Ann. Neurol. 25:351; Howard et al., 1989, J. Neurosurg. 71:105). In another embodiment, a controlled release system can be placed near the target area of ​​treatment, e.g., the retina, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)). Other controlled release systems discussed in the review by Langer, 1990, Science 249:1527-1533, can also be used.

[0161] Administration of any of the agents described herein can independently be 1 to 4 times daily, 1 to 4 times monthly, 1 to 6 times per year, or once every 2, 3, 4, or 5 years. Administration can be daily or for a period of 1, 2, 3, 6, 1, 2, or 3 years, or even for the lifetime of the subject. Long-term administration is often indicated. The dosage can be administered as a single dose or divided into multiple doses. Generally, the desired dosage will be administered at set intervals over an extended period of time, usually for at least several weeks or months, although longer administration periods of several months or years or longer may be required.

[0162] The dosage regimen of any of the drugs described herein can be selected according to various factors, such as the type, species, age, weight, sex and condition of the subject; the severity of the condition to be treated; the route of administration; the renal or hepatic function of the subject; the pharmacogenomic makeup of the individual and the specific compound of the present disclosure used.Any of the drugs described herein can be administered in a single daily dose, or the total daily dosage can be administered in divided doses 2, 3 or 4 times a day.In addition, any of the drugs described herein can be administered continuously throughout the entire dosage regimen, not intermittently.

[0163] Subjects and / or animals In some embodiments, the subject and / or animal is a mammal, such as a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, rabbit, sheep, or non-human primate, such as a monkey, chimpanzee, or baboon. In other embodiments, the subject and / or animal is a non-mammal, such as a zebrafish. In some embodiments, the subject and / or animal may comprise fluorescently tagged cells (e.g., with GFP). In some embodiments, the subject and / or animal is a transgenic animal comprising fluorescent cells. In some embodiments, the subject and / or animal is a human. In some embodiments, the human is a pediatric human. In other embodiments, the human is an adult human. In other embodiments, the human is an elderly human. In other embodiments, the human may be referred to as a patient.

[0164] In certain embodiments, the human has an age ranging from about 0 to about 6 months, about 6 to about 12 months, about 6 to about 18 months, about 18 to about 36 months, about 1 to about 5 years, about 5 to about 10 years, about 10 to about 15 years, about 15 to about 20 years, about 20 to about 25 years, about 25 to about 30 years, about 30 to about 35 years, about 35 to about 40 years, about 40 to about 45 years, about 45 to about 50 years, about 50 to about 55 years, about 55 to about 60 years, about 60 to about 65 years, about 65 to about 70 years, about 70 to about 75 years, about 75 to about 80 years, about 80 to about 85 years, about 85 to about 90 years, about 90 to about 95 years, or about 95 to about 100 years.

[0165] In other aspects, the subject is a non-human animal, and thus the present disclosure relates to veterinary uses. In one particular aspect, the non-human animal is a domestic pet. In another particular aspect, the non-human animal is a livestock animal.

[0166] kit The present disclosure provides a kit that can simplify the administration of any of the drugs described herein.The exemplary kit of the present disclosure comprises any of the drugs described herein in unit dosage form.In one embodiment, the unit dosage form is a container, such as a pre-filled syringe, that can be sterile and comprises any of the drugs described herein and pharmaceutically acceptable carriers, diluents, excipients or vehicles.The kit can also comprise a label or printed instruction manual for teaching the use of any of the drugs described herein.The kit can also comprise a cap-type speculum, a topical anesthetic and a cleaning agent for the ocular surface.The kit can also comprise one or more additional drugs described herein.

[0167] In one aspect, the kit comprises a container containing an effective amount of an agent of the disclosure, such as a compound of Formula I, Ia, II, III, IV, V, VI, or VII or a pharmaceutically acceptable salt thereof, or any compound disclosed herein, and an effective amount of another therapeutic agent, such as described herein.

[0168] definition The following definitions are used in the context of the present disclosure disclosed herein: Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0169] An "effective amount," when used in the context of an agent of the disclosure, e.g., a BCAT2 inhibitor (e.g., a compound of Formula I, Ia, II, III, IV, V, VI, or VII or a pharmaceutically acceptable salt thereof, or any compound disclosed herein), is an amount effective to reduce the toxic burden of BCAA metabolites and / or substrate molecules as described herein.

[0170] An agent is "useful in the treatment of organic acidemia" if the agent results in a measurable treatment of the organic acidemia, inhibiting its etiology or reducing its rate and / or symptoms.

[0171] As used herein, "a," "an," or "the" can mean one or more than one. Furthermore, the term "about," when used in connection with a referenced numerical indication, means the referenced numerical indication plus or minus up to 10% of the referenced numerical indication. For example, the phrase "about 50" encompasses a range of 45 to 55.

[0172] When referred to herein, all percentages of compositions are based on the weight of the entire composition unless otherwise specified.As used herein, the word "comprises" and its variants are intended to be non-limiting, and therefore the description of an object in a list does not exclude other similar objects that may also be useful in the materials, compositions, devices and methods of the present technology.Similarly, the terms "can / can" and "may / can" and their variants are intended to be non-limiting, and therefore the description that an embodiment may or may include some specific elements or features does not exclude other embodiments of the present technology that do not include those elements or features.

[0173] Although the open-ended term "comprising" is used synonymously with terms such as including, containing, or having to describe and claim the disclosure herein, the disclosure or embodiments thereof may alternatively be described using alternative terms, such as "consisting of" or "consisting essentially of."

[0174] "Solvate" refers to the compound formed by the interaction of a solvent with EPI, its metabolite, or salt. Suitable solvates are pharmaceutically acceptable solvates, such as hydrates.

[0175] As used herein, "a" and "b" are integers. a ~C b " or "C a~b " indicates the number of carbon atoms in the specified group; that is, the group may contain from "a" to "b" carbon atoms, inclusive. Thus, for example, "C1-C4 alkyl" or "C 1~4 An "alkyl" group refers to any alkyl group having 1 to 4 carbons, i.e., CH3-, CH3CH2-, CH3CH2CH2-, (CH3)2CH-, CH3CH2CH2CH2-, CH3CH2CH(CH3)-, and (CH3)3C-.

[0176] The term "halogen" or "halo" as used herein means any one of the radio-stable atoms in column 7 of the periodic table of the elements, e.g., fluorine, chlorine, bromine, or iodine, with fluorine and chlorine being preferred.

[0177] "Alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, and having from 1 to 10 carbon atoms (e.g., (C1-10) alkyl or C1-10 alkyl). Wherever it appears herein, a numerical range, e.g., "1-10," refers to each integer within the range indicated—e.g., "1-10 carbon atoms" means that the alkyl group can be 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms, although this definition is intended to encompass the occurrence of the term "alkyl" without specifically specifying a numerical range. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, isobutyl, tertiary butyl, pentyl, isopentyl, neopentyl, hexyl, septyl, octyl, nonyl, and decyl. Alkyl moieties, such as methyl (Me), ethyl (Et), n-propyl (Pr), 1-methylethyl (iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), and 3-methylhexyl, can be attached to the remainder of the molecule by a single bond.Unless stated otherwise specifically in the specification, an alkyl group is optionally substituted by one or more substituents, which substituents independently include alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, —ORa, —SRa, —OC(O)-Ra, —N(Ra), —C(O)Ra, —C(O)ORa, —OC(O)N(Ra), —C(O)N(Ra), —N( and PO(Ra)2, where each Ra is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0178] "Cycloalkyl" refers to a monocyclic or polycyclic group of atoms that contains only carbon and hydrogen and can be saturated or partially unsaturated. Cycloalkyl groups include groups having 3 to 10 ring atoms (i.e., (C3-10)cycloalkyl or C3-10cycloalkyl). Wherever it appears herein, numerical ranges, e.g., "3 to 10," refer to each integer within the range indicated—e.g., "3 to 10 carbon atoms" means that the cycloalkyl group can consist of up to and including 10 carbon atoms, such as 3 carbon atoms. Illustrative examples of cycloalkyl groups include, but are not limited to, the following moieties: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloseptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl, and the like. Unless stated otherwise specifically in the specification, a cycloalkyl group is optionally substituted by one or more substituents, which substituents are independently alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, ═O, —ORa, —SRa, —OC(O)—Ra, —N(Ra), —C(O)Ra, —C(O)ORa, —OC(O)N(Ra), —C(O)N(Ra), -N(Ra)C(O)ORa, -N(Ra)C(O)Ra, -N(Ra)C(O)N(Ra), N(Ra)C(NRa)N(Ra), -N(Ra)S(O)tRa (where t is 1 or 2), -S(O)tORa (where t is 1 or 2), -S(O)tN(Ra) (where t is 1 or 2) or PO(Ra), where each Ra is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0179] As used herein, "haloalkyl" refers to a straight- or branched-chain alkyl group having 1 to 12 carbon atoms in the chain and in which one or more hydrogens have been replaced with a halogen. Examples of haloalkyl groups include, but are not limited to, -CF, -CHF, -CHF, -CHCF, -CHCHF, -CHCHF, -CHCHCl, -CHCFCF, and other groups that would be considered equivalent to any one of the foregoing examples given one of ordinary skill in the art and the teachings provided herein.

[0180] As used herein, "alkoxy" refers to a group of the formula -OR, where R is alkyl as defined above, e.g., "C 1~9 "Alkoxy" refers to alkoxy groups, and includes, but is not limited to, methoxy, ethoxy, n-propoxy, 1-methylethoxy (isopropoxy), n-butoxy, iso-butoxy, sec-butoxy, and tert-butoxy.

[0181] As used herein, "aryl" refers to an aromatic ring or ring system (i.e., two or more fused rings that share two adjacent carbon atoms) that contains only carbon in the ring backbone. When aryl is a ring system, every ring in the system is aromatic. An aryl group can have 6 to 18 carbon atoms, although this definition also encompasses the occurrence of the term "aryl" without specifying a numerical range. In some embodiments, an aryl group has 6 to 10 carbon atoms. An aryl group is defined as "C 6~10 Aryl, C6 to C 10 The aryl group may be designated as "aryl" or similar designations. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, azulenyl, and anthracenyl.

[0182] As used herein, "aryloxy" and "arylthio" refer to RO- and RS-, where R is aryl as defined above, e.g., "C 6~10 aryloxy" or "C 6~10 and the like, including, but not limited to, phenyloxy.

[0183] "Aralkyl" or "arylalkyl" refers to the "C 7~14 Aryl groups linked as substituents via alkylene groups, such as "aralkyl" and the like, include, but are not limited to, benzyl, 2-phenylethyl, 3-phenylpropyl, and naphthylalkyl. In some cases, the alkylene group may be a lower alkylene group (i.e., C 1~4 alkylene group).

[0184] As used herein, "heteroaryl" refers to an aromatic ring or ring system (i.e., two or more fused rings sharing two adjacent atoms) containing one or more heteroatoms, i.e., elements other than carbon, such as, but not limited to, nitrogen, oxygen, and sulfur, within the ring backbone. When heteroaryl is a ring system, every ring in the system is aromatic. Heteroaryl groups can have 5 to 18 ring members (i.e., the number of atoms, such as carbon atoms and heteroatoms, that comprise the ring backbone), although this definition also encompasses the existence of the term "heteroaryl" without specifying a numerical range. In some embodiments, heteroaryl groups have 5 to 10 ring members or 5 to 7 ring members. Heteroaryl groups can be designated as "5-7-membered heteroaryl," "5-10-membered heteroaryl," or similar notations. In various embodiments, heteroaryls contain 1 to 4 heteroatoms, 1 to 3 heteroatoms, 1 to 2 heteroatoms, or 1 heteroatom. For example, in various embodiments, heteroaryl contains 1 to 4 nitrogen atoms, 1 to 3 nitrogen atoms, 1 to 2 nitrogen atoms, 2 nitrogen atoms and 1 sulfur or oxygen atom, 1 nitrogen atom and 1 sulfur or oxygen atom, or 1 sulfur or oxygen atom. Examples of heteroaryl include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzoindolyl, 1,3-benzodioxolyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzoxazolyl, benzopyranyl, and benzopyranonyl. benzoyl, benzofuranyl, benzofuranonyl, benzofurazanyl, benzothiazolyl, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-Dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furazanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imino Dazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H -pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinolinyl Quinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, thiapyranyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyridinyl and thiophenyl (i.e., thienyl). Unless specifically stated otherwise in the specification, the heteroaryl moiety can be optionally substituted by one or more substituents, which substituents are independently alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, -ORa, -SRa, -OC(O)-Ra, -N(Ra), -C(O)Ra, -C(O)ORa, -OC(O)N(Ra), -C(O)N(Ra), -N(Ra)C (O)ORa, -N(Ra)C(O)Ra, -N(Ra)C(O)N(Ra), N(Ra)C(NRa)N(Ra), -N(Ra)S(O)tRa (where t is 1 or 2), -S(O)tORa (where t is 1 or 2), -S(O)tN(Ra) (where t is 1 or 2) or PO3(Ra), where each Ra is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0185] Substituted heteroaryl also includes ring systems substituted with one or more oxide (—O—) substituents, such as pyridinyl N-oxides.

[0186] As used herein, "heteroalkyl" refers to a straight or branched hydrocarbon chain containing one or more heteroatoms, i.e., elements other than carbon, such as, but not limited to, nitrogen, oxygen, and sulfur, in the backbone. Heteroalkyl groups can have 1 to 20 carbon atoms, although this definition also encompasses the term "heteroalkyl" without specifying a numerical range. Heteroalkyl groups can also be medium-sized heteroalkyls having 1 to 9 carbon atoms. Heteroalkyl groups could also be lower heteroalkyls having 1 to 4 carbon atoms. In various embodiments, heteroalkyls can have 1 to 4 heteroatoms, 1 to 3 heteroatoms, 1 or 2 heteroatoms, or 1 heteroatom. The heteroalkyl group of a compound is defined as "C 1~4 A heteroalkyl group may be designated "heteroalkyl" or similar designations. A heteroalkyl group may contain one or more heteroatoms. By way of example only, "C 1~4 "Heteroalkyl" indicates that there are from 1 to 4 carbon atoms in the heteroalkyl chain and one or more heteroatoms located within the backbone of the chain.

[0187] As used herein, "carbocyclyl" refers to a non-aromatic ring or ring system containing only carbon atoms within the ring system backbone. When a carbocyclyl is a ring system, two or more rings may be joined together in a fused, bridged, or spiro-type connection. A carbocyclyl may have any degree of saturation, provided that at least one ring in the ring system is not aromatic. Thus, carbocyclyl includes cycloalkyl, cycloalkenyl, and cycloalkynyl. A carbocyclyl group may have 3 to 20 carbon atoms, although this definition also encompasses the term "carbocyclyl" without specifying a numerical range. A carbocyclyl group may also be a medium-sized carbocyclyl having 3 to 10 carbon atoms. A carbocyclyl group could also be a carbocyclyl having 3 to 6 carbon atoms. A carbocyclyl group is defined as a "C 3~6Carbocyclyl" or similar designations. Examples of carbocyclyl rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, 2,3-dihydro-indene, bicycle[2.2.2]octanyl, adamantyl, and spiro[4.4]nonanyl.

[0188] "(Carbocyclyl)alkyl" refers to a carbocyclyl group linked as a substituent via an alkylene group, e.g., "C 4~10 (carbocyclyl)alkyl" and the like, including, but not limited to, cyclopropylmethyl, cyclobutylmethyl, cyclopropylethyl, cyclopropylbutyl, cyclobutylethyl, cyclopropylisopropyl, cyclopentylmethyl, cyclopentylethyl, cyclohexylmethyl, cyclohexylethyl, cycloheptylmethyl, and the like. In some cases, the alkylene group is a lower alkylene group.

[0189] As used herein, "heterocyclyl" refers to a non-aromatic ring or ring system containing at least one heteroatom within the ring backbone. Heterocyclyls can be joined together in fused, bridged, or spiro-type linkages. Heterocyclyls can have any degree of saturation, provided that at least one ring in the ring system is not aromatic. The heteroatom(s) can be present in either a non-aromatic or aromatic ring within the ring system. Heterocyclyl groups can have 3 to 20 ring members (i.e., the number of atoms, such as carbon atoms and heteroatoms, comprising the ring backbone), although this definition also encompasses the term "heterocyclyl" without specifying a numerical range. Heterocyclyl groups can also be medium-sized heterocyclyls having 3 to 10 ring members. Heterocyclyl groups could also be heterocyclyls having 3 to 6 ring members. Heterocyclyl groups can be designated as "3- to 6-membered heterocyclyl" or similar notations.

[0190] In various embodiments, the heterocyclyl contains 1 to 4 heteroatoms, 1 to 3 heteroatoms, 1 to 2 heteroatoms, or 1 heteroatom. For example, in various embodiments, the heterocyclyl contains 1 to 4 nitrogen atoms, 1 to 3 nitrogen atoms, 1 to 2 nitrogen atoms, 2 nitrogen atoms and 1 sulfur or oxygen atom, 1 nitrogen atom and 1 sulfur or oxygen atom, or 1 sulfur or oxygen atom. In preferred 6-membered monocyclic heterocyclyls, the heteroatom(s) are selected from 1 to 3 O, N, or S, and in preferred 5-membered monocyclic heterocyclyls, the heteroatom(s) are selected from 1 or 2 heteroatoms selected from O, N, or S. Examples of heterocyclyl rings include, but are not limited to, azepinyl, acridinyl, carbazolyl, cinnolinyl, dioxolanyl, imidazolinyl, imidazolidinyl, morpholinyl, oxiranyl, oxepanyl, thiepanyl, piperidinyl, piperazinyl, dioxopiperazinyl, pyrrolidinyl, pyrrolidonyl, pyrrolidionyl, 4-piperidonyl, pyrazolinyl, pyrazolidinyl, 1,3-dioxinyl, 1,3-dioxanyl, 1,4-dioxinyl, 1,4-dioxanyl, 1,3-oxathianyl, 1,4-oxathiinyl, 1,4-oxathianyl, 2H-1,2-oxazinyl, trioxanyl, Examples include hexahydro-1,3,5-triazinyl, 1,3-dioxolyl, 1,3-dioxolanyl, 1,3-dithiolyl, 1,3-dithiolanyl, isoxazolinyl, isoxazolidinyl, oxazolinyl, oxazolidinyl, oxazolidinonyl, thiazolinyl, thiazolidinyl, 1,3-oxathiolanyl, indolinyl, isoindolinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydro-1,4-thiazinyl, thiamorpholinyl, dihydrobenzofuranyl, benzimidazolidinyl, and tetrahydroquinoline.

[0191] "(Heterocyclyl)alkyl" is a heterocyclyl group linked as a substituent via an alkylene group. Examples include, but are not limited to, imidazolinylmethyl and indolinylethyl.

[0192] As used herein, "acyl" refers to -C(=O)R, where R is hydrogen, C(=O) as defined herein. 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 carbocyclyl, aryl, 5-10 membered heteroaryl and 5-10 membered heterocyclyl. Non-limiting examples include formyl, acetyl, propanoyl, benzoyl and acryl.

[0193] An "O-carboxy" group refers to an "-OC(=O)R" group, where R is hydrogen, C, as defined herein. 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 It is selected from carbocyclyl, aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0194] A "C-carboxy" group refers to a "-C(=O)OR" group, where R is hydrogen, C(=O)OR, as defined herein. 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 It is selected from carbocyclyl, aryl, 5-10 membered heteroaryl and 5-10 membered heterocyclyl. A non-limiting example is carboxyl (i.e., -C(=O)OH).

[0195] A "cyano" group refers to a "-CN" group.

[0196] A "cyanato" group refers to an "-OCN" group.

[0197] An "isocyanato" group refers to an "-NCO" group.

[0198] A "thiocyanato" group refers to a "-SCN" group.

[0199] An "isothiocyanato" group refers to an "-NCS" group.

[0200] A "sulfinyl" group refers to a "-S(=O)R" group, where R is hydrogen, C, as defined herein. 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 Carbocyclyl, C 6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0201] A "sulfonyl" group refers to a "-SO2R" group, where R is hydrogen, C, as defined herein. 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 Carbocyclyl, C 6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0202] The "S-sulfonamide" group is "-SO2NR A R B " group, where R A and R B are each independently hydrogen, C as defined herein 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 Carbocyclyl, C 6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0203] The "N-sulfonamide" group is "-N(R A )SO2R B " group, where R A and R b are each independently hydrogen, C as defined herein 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 Carbocyclyl, C6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0204] The "O-carbamyl" group is "-OC(=O)NR A R B " group, where R A and R B are each independently hydrogen, C as defined herein 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 Carbocyclyl, C 6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0205] The "N-carbamyl" group is "-N(R A )OC(=O)R B " group, where R A and R B are each independently hydrogen, C as defined herein 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 Carbocyclyl, C 6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0206] The "O-thiocarbamyl" group is "-OC(=S)NR A R B " group, where R A and R B are each independently hydrogen, C as defined herein 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 Carbocyclyl, C 6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0207] The "N-thiocarbamyl" group is "-N(R A)OC(=S)R B " group, where R A and R B are each independently hydrogen, C as defined herein 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 Carbocyclyl, C 6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0208] The "C-amide" group is "-C(=O)NR A R B " group, where R A and R B are each independently hydrogen, C as defined herein 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 Carbocyclyl, C 6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0209] The "N-amide" group is "-N(R A )C(=O)R B " group, where R A and R B are each independently hydrogen, C as defined herein 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~7 Carbocyclyl, C 6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0210] The "amino" group is "-NR A R B " group, where R A and R B are each independently hydrogen, C as defined herein 1~6 Alkyl, C 2~6 Alkenyl, C2~6 Alkynyl, C 3~7 Carbocyclyl, C 6~10 It is selected from aryl, 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclyl.

[0211] An "aminoalkyl" group refers to an amino group linked via an alkylene group.

[0212] An "alkoxyalkyl" group is an alkoxy group linked via an alkylene group, e.g., "C 2~8 "Alkoxyalkyl" and the like.

[0213] It will be understood that some specific moiety naming conventions may include either monovalent or divalent moieties, depending on the context. For example, if a substituent requires two points of attachment to the remainder of the molecule, it will be understood that the substituent is a divalent moiety. For example, substituents identified as alkyl requiring two points of attachment include divalent moieties such as -CH-, -CHCH-, -CHCH(CH)CH-, etc. Other moiety naming conventions specify that the moiety is a divalent moiety, such as "alkylene" or "alkenylene."

[0214] When two R groups are said to "together with the atom(s) to which they are attached" to form a ring (e.g., a carbocyclyl, heterocyclyl, aryl, or heteroaryl ring), it is meant that the atoms and the two R groups together form the described ring. The ring is not otherwise limited by the definition of each R group when taken individually. For example, the following substructure exists: TIFF0007789565000168.tif19128R 1 and R 2 is defined as being selected from the group consisting of hydrogen and alkyl, or R 1 and R 2 When these are joined together with the nitrogen to which they are attached to form a heterocyclyl, R 1 and R 2may be selected from hydrogen or alkyl, or alternatively, the moiety may have the structure: TIFF0007789565000169.tif14128, where Ring A is the depicted nitrogen-containing heterocyclyl ring.

[0215] Similarly, when two "adjacent" R groups are said to "together with the atom to which they are attached" to form a ring, it means that the atom, the intervening bond, and the collective unit of the two R groups is the described ring. For example, the following substructure exists: TIFF0007789565000170.tif26128R 1 and R 2 is defined as being selected from the group consisting of hydrogen and alkyl, or R 1 and R 2 When these together with the atom to which they are attached form an aryl or carbocyclyl, R 1 and R 2 may be selected from hydrogen or alkyl, or alternatively, the moiety may have the structure: TIFF0007789565000171.tif26128, where A is an aryl ring or carbocyclyl containing the double bond shown.

[0216] Whenever a substituent is depicted as a divalent group (i.e., having two points of attachment to the rest of the molecule), it is understood that the substituent may be attached in any orientation unless otherwise stated. Thus, for example, -AE- or The substituent depicted as TIFF0007789565000172.tif8128 includes that substituent oriented such that A is attached to the point of attachment at the left end of the molecule as well as when A is attached to the point of attachment at the right end of the molecule.

[0217] The present disclosure is further illustrated by the following non-limiting examples. [Example]

[0218] Example 1: In vitro inhibition of BCAT2 and BCAT1 In this example, a compound of the present disclosure, a BCAT2 inhibitor compound of formula (I) shown below: Whether TIFF0007789565000173.tif37128 can inhibit the levels of the downstream metabolite propionylcarnitine, which is found in elevated levels in patients with propionic acidemia, was evaluated by analyzing propionylcarnitine levels in the conditioned medium of human fibroblasts derived from patients with propionic acidemia treated with a response dose of a compound of the present disclosure.

[0219] Primary human fibroblasts (Coriell #GM00371) derived from a patient with propionic acidemia were plated at 10,000 cells / well in complete medium (EMEM, 15% FBS) in a 96-well plate and incubated overnight at 37°C with 5% CO2. Compounds were serially diluted first in DMSO and then in serum-free EMEM medium. Cells were then washed with D-PBS, after which medium containing the diluted compounds was added and then incubated overnight at 37°C with 5% CO2.

[0220] After 24 hours of compound treatment, conditioned medium was removed from the cells and diluted twice with D3-propionylcarnitine internal standard diluted in MS-grade methanol. The plate was sealed, and the samples were frozen at -80°C for >30 minutes, then thawed at room temperature and centrifuged at 4000 rpm for 10 minutes. The samples were then analyzed for propionylcarnitine by LC-LC / MS using a Kinetex 2.6 μm Biphenyl 2.1 × 50 mm column on a TSQ Vantage with a mobile phase A of 95% (water with 0.1% formic acid) + 5% water and a mobile phase B of 95% MeOH + 5% (water with 0.1% formic acid).

[0221] Cell viability was also measured using cell Titer-Glo reagent (Promega) by adding equal volumes of D-PBS and Cell Titer-Glo reagent to wells of cells and following the manufacturer's recommendations for detection of the luminescent signal.

[0222] The results of the experiment are shown in Figure 1, which demonstrates that the BCAT2 inhibitor is a compound of formula Ia, shown below: Figure 3 shows the effect of TIFF0007789565000174.tif37128 on BCAA pathway flux and on the downstream metabolite propionylcarnitine. The results show that in the presence of increasing concentrations of BCAT2 inhibitors, propionylcarnitine levels are inhibited with little effect on cell viability, suggesting that BCAT2 reduces BCAA pathway flux.

[0223] The BCAT1 inhibitory activity of selected compounds of the present disclosure was also evaluated. Compounds were generally prepared as stock solutions up to 10 mM using DMSO as a vehicle. 10-point dose-response curves were generated using an Echo-550 (Labcyte) acoustic dispenser. Compound source plates (384-well, opaque white assay plates, Corning #3570) were prepared by serially diluting compound stock solutions to create 1 mM, 0.1 mM, and 0.01 mM solutions in DMSO in Echo-certified LDV plates. 100% DMSO stock solutions were then serially spotted into a dose-response source plate using Echo, creating a semi-logarithmic dilution scheme. 100% DMSO was added to the spotted dose-response plate to a final volume of 5 μL to create a 333× stock plate. 120 nL of the dose-response stock plate was then spotted into an assay plate. To the plate was then added 10 μL of assay buffer (50 mM Tris with 0.05% TWEEN-20) to bring the assay plate to 4× working concentration, with a final assay test concentration range of 3 μM to 0.0001 μM and a final DMSO concentration of 0.3%.

[0224] The BCAT1 enzyme stock solution was first diluted to a 4x working concentration of 0.24µg / mL in assay buffer (resulting in a final assay concentration of 0.06µg / mL). 10µL of the 4x BCAT1 solution was then added to the previously prepared assay plates containing test compounds or controls and incubated at room temperature, protected from light, for either 10 or 60 minutes. After the compound incubation period, 20µL of assay buffer + 2x substrate was added to all wells (final assay concentrations of substrates were 300µM L-leucine and 250µM alpha-ketoglutarate). The enzyme reaction was then incubated at room temperature for 10 minutes. After the 10-minute incubation, the enzyme reaction was stopped by adding 5µL of 0.6N HCl and incubated for 1 minute. 5µL of 1M Tris (pH 8.0) was then added to neutralize the pH. A volume of 12.5 μL of the quench assay was then transferred to a white 384-well plate, and 12.5 μL of glutamate detection reagent was added (prepared as per the instructions in the Promega Glutamate Glo kit). The plate was then incubated at room temperature, protected from light, for 30 minutes. After the glutamate detection incubation period, luminescence was read on a BMG Novostar to measure relative luminescence and export for analysis.

[0225] Modulation of BCAT1 enzyme activity by compounds was measured as follows: IC 50 Values ​​were calculated in ActivityBase using a four-parameter fit equation. Data were normalized to the maximally inhibited reference drug VS558CSA (10 μM). Relative light units were exported for analysis and % inhibition values ​​for each well were calculated using the following formula: Calculations were made using TIFF0007789565000175.tif17128.

[0226] BCAT2 inhibitory activity for additional compounds of the present disclosure is shown below along with BCAT1 inhibition data.

[0227] TIFF0007789565000176.tif175167TIFF0007789565000177.tif219167TIFF0007789565000178.tif132167

[0228] Example 2: BCAT2 In Vivo Inhibition In this example, the BCAT2 inhibitor compound shown below: The pharmacodynamics following acute and repeated treatment with TIFF0007789565000179.tif43128 were evaluated.

[0229] chemicals Branched-chain aminotransferase 2 (BCAT2) inhibitor compounds were synthesized according to Example 3.

[0230] Hydroxypropyl methylcellulose (HPMC) and the branched-chain amino acids L-cysteine, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, L-phenylalanine, L-threonine, L-tryptophan, L-tyrosine, and L-valine were purchased from Sigma-Aldrich (St. Louis, MO). All other reagents were obtained from commercial suppliers and were of reagent grade or higher.

[0231] animal Male C57Black / 6J mice (6-7 weeks old, Jackson Laboratories, Bar Harbor, ME) were used for all in vivo experiments. Animals were maintained on a 12-hour reversed light / dark cycle in a temperature- and humidity-controlled room and had access to food and water ad libitum. Animals were allowed a 7-day acclimation period before experimentation. All procedures involving mice were approved by the University of Arizona Institutional Animal Care and Use Committee and were performed in accordance with the Principles of Laboratory Animal Care (National Institutes of Health Publication 85-23, revised 1985).

[0232] Bioanalysis of plasma, urine, and tissue homogenate extracts All biological samples were stored at -80°C until analysis. Branched-chain amino acids (valine, leucine / isoleucine) and serine in mouse plasma and urine were extracted using a protein precipitation-based method with D8-valine in acetonitrile (ACN) as an internal standard and analyzed by liquid chromatography-mass spectrometry (LC-MS / MS) on an Agilent 1290 Infinite II separation HPLC coupled to an Agilent 6490 QQQ MS. Specifically, 20 μL of mouse plasma was precipitated with 180 μL of acetonitrile containing 5 μg / mL D8-valine. The samples were vigorously vortexed and centrifuged to remove precipitated proteins. Two microliters of the supernatant was injected onto a Phenomenex Gemini C18 column (4.6 mm x 150 mm, 5 μm particle size) using a linear gradient from 2% ACN to 50% ACN over 10 minutes, followed by equilibration at 2% ACN for 7 minutes. The amino acid transitions were 132.1–86.1 for leucine / isoleucine, 106.1–60.1 for serine, and 118.1–72.1 for valine.

[0233] Propionylcarnitine in mouse plasma was extracted by solid-phase extraction (SPE) and analyzed by LC-MS / MS on a Thermo Scientific TSQ vantage. Samples were injected onto a Kinetex Biphenyl column (2.1 x 50 mm, 2.6 μm) maintained at room temperature. Analytes were eluted with a linear gradient consisting of 0.095% formic acid in water (A) and 0.0005% formic acid in MeOH (B).

[0234] Pharmacodynamic studies: acute treatment with BCAT2 inhibitors After overnight fasting, the animals were orally administered vehicle (0.5% HPMC K100 + 0.1 Tween 80, pH = 10.0) or the indicated doses of compounds (30, 100, 300 mg / kg) as a suspension for 6 hours. After compound administration, the animals had free access to food and water. Each mouse was then given an oral feeding mix of 1.5 g / 10 mL / kg of amino acids. The amino acid mix was prepared in 0.5% HPMC / 0.1% Tween 80, pH 7, and contained 114 mg L-cysteine, 114 mg L-histidine, 468 mg L-isoleucine, 939 mg L-leucine, 486 mg L-lysine, 39 mg L-methionine, 75 mg L-phenylalanine, 264 mg L-threonine, 15 mg L-tryptophan, 21 mg L-tyrosine, and 468 mg L-valine. One hour after the amino acid challenge, blood samples were collected by intracardiac puncture using K2-EDTA-coated tubes. Plasma was collected after centrifugation and stored at -80°C until analysis.

[0235] The results shown in Figure 2 indicate that BCAT2 inhibitor compounds can block branched-chain amino acids (BCAAs) from entering their catabolic pathway. As shown in Figure 2, BCAT2 inhibitor compounds increased plasma valine (Val), isoleucine (Ile), and leucine (Leu) levels in a dose-dependent manner compared to vehicle controls. This experiment demonstrated that BCAT2 inhibitor compounds effectively increased BCAA levels in an acute animal model.

[0236] Pharmacodynamic studies: Repeated treatment with BCAT2 inhibitor compounds Animals were grouped based on weight-matched pairs and received either vehicle (0.5% HPMC K100 + 0.1 Tween 80, pH = 10.0) or a daily dose of compound (200 mg / kg, PO) for 9 days. From days 1 to 5, all animals were fed a normal diet. On day 6, half of the animals were switched to a precursor-rich diet (700% isoleucine, valine, and threonine-enriched, TD.140829; Envigo); the other half remained on a normal diet. On day 9, blood, bladder urine, and tissue samples (liver, brain, heart, muscle, and kidney) were collected 4 hours after the last dose. Three mice from each treatment / diet group were retained for a 3-day recovery observation period. On day 12, blood, bladder urine, and tissue samples (liver, brain, heart, muscle, and kidney) were collected. During the entire study period (days 1 to 12), mice were allowed free access to food and water. Body weight was measured daily, and any abnormal behaviors indicating toxicity were recorded.

[0237] Figures 3A-D show the results of monitoring mouse weight during the study. Considering that weight may be a confounding factor in examining BCAA and PC levels in biological samples after repeated dosing, mice were grouped into weight-matched pairs on Day 0 and monitored for weight changes throughout the study. Figure 3A shows that no significant weight loss was observed during the study. No significant differences were observed between groups in weight measured before treatment (Day 0) or on the two collection days (Days 9 and 12) (Figures 3B, 3C, and 3D). Furthermore, no obvious abnormal behaviors reflecting toxicity were observed during the study. In each set of Figures 3B, 3C, and 3D, the left bar represents vehicle, and the right bar represents BCAA.

[0238] Figures 4A-B show plasma levels of BCAAs in mice after repeated treatment with a BCAT2 inhibitor compound. As shown in Figure 4A, repeated dosing with a BCAT2 inhibitor compound (200 mg / kg, PO) for 9 days significantly increased plasma levels of Val and Leu, but not Ile and Ser, in mice fed a normal diet. In each pair, the left bar represents vehicle, and the right bar represents BCAT.

[0239] Changes in catabolic status or excessive protein intake often lead to increased throughput of the propionic acid metabolic pathway in patients with propionic acidemia. To mimic this condition and test whether BCAT2 inhibitor compounds can block BCAAs from entering the propionic acid metabolic pathway, a food challenge was used in a repeated-dose study. Five days after compound treatment, mice were switched to a precursor-rich diet consisting of high levels of isoleucine, valine, and threonine. Data showed no significant changes in BCAA levels in mouse plasma collected on day 9 between compound and vehicle treatment, except for a slight but significant decrease in plasma levels of leucine observed in the day 9 sample (Figure 4B).

[0240] Figures 5A-B show plasma (5A) and urinary (5B) amino acid levels in samples from mice fed either a normal diet or a precursor-rich diet on day 9. Figures 5A-B show that BCAA overload by feeding a precursor-rich diet leads to increased renal excretion of these amino acids.

[0241] Example 3: In vitro 3-HIB assay Cell plating: Cells (Coriell Institute GM00371 cells) were plated at 10,000 cells / 0.1 mL in Eagle's Minimum Essential Medium (EMEM) / 15% FBS (Gibco 16000-044) using a Combi dispenser in a cell culture hood (96-well clear-bottom sterile plates; Greiner 655098) and incubated overnight at 37°C and 5% CO2.

[0242] Compound dilutions: Compounds were prepared as 33.3x intermediate dilutions in DMSO in 96-well polystyrene round plates. 30 μL of the dilution started at 10 mM in DMSO for a final maximum of 30 μM. 10-12 1:3 serial dilutions were prepared from left to right (20 μL + 40 μL DMSO). 10x secondary dilutions in EMEM were prepared by transferring 7.5 μL of the 33.3x dilution to a replicate 96-well polystyrene round plate and then adding 242.5 μL of EMEM. 1x final dilutions in EMEM were prepared by transferring 50 μL of the 10x dilution to a replicate 96-well polypropylene assay block (Costar 3956) and adding 450 μL of EMEM. A compound maximum inhibition control was prepared by taking 20 μL of 10 mM reference compound A and adding 40 μL of DMSO to a concentration of 3.3 mM. To 7.5 μL of a 3.3 mM solution of reference compound A, 242.5 μL of EMEM was added to make a 100 μM solution. This reference compound A solution was then diluted with EMEM to a final concentration of 10 μM. The structure of reference compound A is The file is TIFF0007789565000180.tif34128.

[0243] A control for maximum compound toxicity was prepared by taking 7.5 μL of Reference Compound B and 242.5 μL of EMEM to make a 300 μM solution. 50 μL of this 100 μM Reference Compound B solution was combined with 450 μL of EMEM to give a concentration of 30 μM. The structure of Reference Compound B is The file is TIFF0007789565000181.tif27128.

[0244] A DMSO control was prepared by diluting 7.5 μL with 242.5 μL of EMEM, then taking 50 μL of this solution and diluting it with 450 μL of EMEM.

[0245] Compound treatment: The medium was aspirated from the cells, 100 μL of D-PBS (ThermoFisher 14190144) was added, and then removed. 100 μL of diluted compound was added to the cells in situ, and the cells were incubated at 37° C. and 5% CO for 24 hours.

[0246] Conditioned Medium Collection: For 3HIB MS detection: 20 μL of conditioned medium was removed from the cells and transferred to a labeled, identical 96-well polypropylene assay plate (VWR 62408-946), which was then heat-sealed and stored at -80°C.

[0247] Viability assay: Cells remaining in the plate were added to 25 μL of D-PBS and 25 μL of Cell titer glo reagent (Promega G7572). Plates were placed on a shaker for 2 minutes, incubated in the dark at room temperature, and then read on a Clariostar microplate reader.

[0248] Derivatization of 3HIB (Generalized SOP): Media collected for 3HIB MS detection was derivatized under the following conditions. Buffer: 0.1 M 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) dissolved in 0.1 M 2-(N-morpholino)ethanesulfonic acid (MES) adjusted to pH 5.6. Buffer preparation: Weigh out 10.6 g of MES (Sigma #M-5287) per 500 mL of HPLC-grade HO. Store at 4°C. 0.1 M EDC: Weigh out 1.971 g of EDC (#E-1769) per 100 mL of MES. Adjust the pH to 5.6 with approximately 40 drops of 1 N NaOH and store at 4°C. The buffer has a short half-life and should be used within 2 weeks.

[0249] Derivatization: 20 μL of sample was added with 60 μL of 5 μM 3HIB-4C13 internal standard, followed by 20 μL of EDC-containing MES and 100 μL of MES. The plate was sealed and incubated at 50° C. for w hours, then centrifuged at 4000 rpm for 10 minutes at 6° C. LC-MS analysis was then performed and data were processed using standard methods.

[0250] TIFF0007789565000182.tif49128TIFF0007789565000183.tif218170TIFF0007789565000184.tif19684

[0251] Example 3: Synthesis of BCAT2 inhibitor compounds Compound synthesis All solvents and reagents were used as received from commercial sources. H NMR spectra were recorded on a Varian Mercury 400 Plus. Chemical shifts are expressed in parts per million (ppm, δ units). Coupling constants (J) are in hertz (Hz). Splitting patterns indicate apparent multiplicity and are designated as s (singlet), d (doublet), t (triplet), q (quartet), dd (double doublet), dt (double triplet), m (multiplet), and br (broad). Analytical purity of final compounds is ≥95% unless otherwise noted. Purity of final compounds was confirmed using an Agilent 1100 HPLC system coupled to a Thermo Finnigan LCQ mass spectrometer. All mass spectra were obtained by electrospray ionization (ESI). Compound purity was analyzed using two different HPLC conditions: LC-MS Method A: 10–98% AcCN-H2O (0.1% TFA) in 2.7 min, hold at 98% AcCN for 0.38 min, flow rate 0.9 mL / min on a Phenomenex Luna 3μ C8(2) 100A 30 × 3.00 mm column; LC-MS Method B: 10–95% AcCN-H2O (0.1% formic acid) in 3.0 min, flow rate 0.5 mL / min on a Kinetex 2.6μ C18 100A 30 × 2.10 mm column. High-resolution mass spectrometry (HRMS) was performed on a Waters qTOF Premiere mass spectrometer operating at a resolution of approximately 15,000 in W mode with positive ionization. Injections were performed using a Waters Nanoacquity LC. The HRMS tolerance is 3 mDa or 5 ppm, but most analyses were observed within 0.5 mDa, and the isotopes matched well with the proposed structures. Purification of the final compounds for biological testing was performed on a Gilson GX-281 system with a Phenomenex Luna 5µC8(2) 100 x 21.20 mm 100A column, separated at a flow rate of 22 mL / min using a gradient of 5-95% MeCN / HO (+0.1% TFA or 0.1% formic acid) over 15-20 min.

[0252] tert-Butyl (3-((4-(methylcarbamoyl)-2-nitrophenyl)amino)cyclohexyl)carbamate (3) TIFF0007789565000185.tif25128 To a solution of 4-fluoro-3-nitrobenzoic acid (1.00 g, 5.40 mmol) in acetonitrile (40 mL) was added HATU (2.054 g, 5.40 mmol) and DIPEA (1.03 mL, 5.94 mmol). The reaction was stirred at room temperature for 15 minutes, after which methanamine (HCl salt, 0.401 g, 5.94 mmol) and DIPEA (1.03 mL, 5.94 mmol) were added. The reaction was stirred for 3 hours and then concentrated in vacuo. The residue was dissolved in water and extracted with ethyl acetate. The organic layer was washed with water, dried over Na2SO4, and concentrated in vacuo to give 4-fluoro-N-methyl-3-nitrobenzamide as crude product. This crude product was dissolved in ethanol (40 mL), followed by the addition of tert-butyl (3-aminocyclohexyl)carbamate (1.10 g, 5.13 mmol, racemic mixture of diastereomers) and DIPEA (1.03 mL, 5.94 mmol). The reaction was stirred at 85 °C for 6 h, then concentrated, and the residue was dissolved in ethyl acetate and washed with water. The organic layer was dried over Na2SO4 and condensed in vacuo to give the title compound as crude product in >95% purity (mixture of diatereomers). LC-MS (ESI) m / z [M+1] + =392.7, Rt=2.13 min (HPLC method A).

[0253] tert-Butyl (3-(5-(methylcarbamoyl)-2-(2-(methylthio)phenyl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)carbamate (4) To a solution of 4-fluoro-N-methyl-3-nitrobenzamide (69 mg, 0.35 mmol) in 1,4-dioxane (4 mL) was added 1,1-dimethylethyl (3-aminocyclohexyl)carbamate (54 mg, 0.25 mmol) and DIPEA (87 μL, 0.5 mmol). The reaction was stirred at 80 °C for 24 h, then cooled to room temperature, and 2-(methylthio)benzaldehyde (38 mg, 0.25 mmol), sodium dithionite (131 mg, 0.75 mmol), and water (1.0 mL) were added. The reaction was stirred at 80 °C for 24 h and then concentrated in vacuo. The residue was purified by preparative HPLC to give the title compound 4 (48.9 mg, 39.5% yield). LC-MS(ESI)m / z[M+1] + =495.2; Rt=1.97 min (HPLC method A).

[0254] cis 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (8b) To a solution of crude material 3 (5.4 mmol) in 1,4-dioxane (40 mL) was added picolinaldehyde (0.579 g, 5.40 mmol), sodium dithionite (NaSO) (5.64 g, 32.4 mmol), and water (10 mL). The reaction was stirred at 80 °C for 18 h, then concentrated in vacuo and extracted with ethyl acetate. The ethyl acetate layer was dried over NaSO and concentrated in vacuo. The residue was purified by chromatography (silica gel column, UV detection at 298 nm, elution system: 0-8% CHOH-CHCl over 40 min at a flow rate of 60 mL / min) to give the intermediate tert-butyl (3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)carbamate as a diastereomeric mixture (1.5 g, 62% yield over three steps from 4-fluoro-3-nitrobenzoid acid). LC-MS (ESI) m / z [M+1] + =450.0; Rt=1.78 min (HPLC method A).

[0255] To the above intermediate (1.5 g, 3.34 mmol) in DCM (30 mL) was added TFA (2.57 mL, 33.4 mmol) dropwise. The reaction was stirred at room temperature for 18 hours. The reaction was concentrated in vacuo, and the resulting residue was dissolved in acetonitrile (30 mL) to give Solution A. In a separate vial, a reaction of 5-bromothiophene-2-carboxylic acid (0.691 g, 3.34 mmol), HATU (1.269 g, 3.34 mmol), and DIPEA (1.276 mL, 7.34 mmol) in acetonitrile (30 mL) was stirred at room temperature for 30 minutes. Solution A was added to the reaction. The mixture was stirred at room temperature for 4 hours and then concentrated in vacuo. The residue was taken up in ethyl acetate and water. The aqueous layer was extracted twice with ethyl acetate, and the combined ethyl acetate solution was washed with saturated NaHCO3 solution followed by brine, then dried over Na2SO4 and concentrated to give the crude product. This crude product was first purified by silica gel chromatography (ISCO system, detection at 298 nm, elution with 0–8% CHOH-CHCl over 50 min at a flow rate of 85 mL / min) to give the trans-isomer (800 mg) and a mixture of cis- and trans-isomers (857 mg). The total yield of cis- and trans-isomers was 1.65 g (92% yield over two steps). The 857 mg mixture of cis- and trans-isomers was further purified by preparative HPLC (column: Phenomenex Gemini C18 110A, A x 1A, 100 x 30.00 mm, 5µ column; flow rate: 40 mL / min; UV detection: 254 nm; gradient: 20–55% AcCN-HO (with 0.2% formic acid as a modifier) ​​over 20 min) to give the pure cis-isomer product (8b, 305 mg). MS(ESI) m / z[M+1] + = 538.1 and 540.1; Rt = 2.24 min (HPLC Method B); TIFF0007789565000188.tif43160For the trans-isomer, TIFF0007789565000189.tif50160

[0256] cis 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(2-(methylthio)phenyl)-1H-benzo[d]imidazole-5-carboxamide (1) The title compound was prepared similarly to the synthesis of 8b using TIFF0007789565000190.tif591284 as the starting material in 14% overall yield as a white solid. TIFF0007789565000191.tif44160

[0257] cis 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-phenyl-1H-benzo[d]imidazole-5-carboxamide (8a) TIFF0007789565000192.tif53128 The title compound as a white solid was prepared similarly to the synthesis of 8b in 12.5% ​​overall yield. TIFF0007789565000193.tif39160

[0258] cis 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (8c) TIFF0007789565000194.tif54159 The title compound as a white solid was prepared similarly to the synthesis of 8b in 4.0% overall yield. TIFF0007789565000195.tif44160

[0259] cis 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-3-yl)-1H-benzo[d]imidazole-5-carboxamide (8d) TIFF0007789565000196.tif35128 The title compound as a white solid was prepared in 5 steps (from 4-fluoro-3-nitro-benzoic acid) in 4.6% overall yield similarly to the synthesis of 8b. TIFF0007789565000197.tif50160

[0260] cis 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-4-yl)-1H-benzo[d]imidazole-5-carboxamide (8e) TIFF0007789565000198.tif48128 The title compound as a white solid was prepared in 5 steps (from 4-fluoro-3-nitro-benzoic acid) in 4.0% overall yield similarly to the synthesis of 8b. TIFF0007789565000199.tif43160

[0261] cis N-(3-aminocyclohexyl)-5-bromothiophene-2-carboxamide (6, TFA salt) To a solution of 5-bromothiophene-2-carboxylic acid (19.27 g, 93 mmol) in DCM (200 mL) was added HOBt (16.29 g, 106 mmol), EDCI (20.40 g, 106 mmol), and triethylamine (29.7 mL, 213 mmol). The mixture was stirred at room temperature for 30 minutes. tert-Butyl (3-aminocyclohexyl)carbamate (19.0 g, 89 mmol) was then added. The reaction was stirred at room temperature overnight. The precipitate was filtered and washed with DCM to give the cis-compound, cis-tert-butyl-3-(5-bromothiophene-2-carboxamido)cyclohexyl)carbamate (a mixture of two cis-isomers, 12.0 g, 29.8 mmol, 33.6% yield) as a white solid (the filtrate contained mainly the trans isomer). To the above Boc-protected intermediate cis-tert-butyl-3-(5-bromothiophene-2-carboxamido)cyclohexyl)carbamate (12.0 g, 29.8 mmol) in DCM (100 mL) was added TFA (22.10 mL, 298 mmol). The mixture was stirred at room temperature for 4 hours and then concentrated in vacuo to give a brown viscous oil. This oil was triturated with diisopropyl ether to give the title compound 6 (12.0 g, 97% yield) as a beige powder. LC-MS (ESI) m / z = 304.9 (M+H). + .

[0262] cis 5-Bromo-N-3-((4-(methylcarbamoyl)-2-nitrophenyl)amino)cyclohexyl)thiophene-2-carboxamide (7) To a solution of 4-fluoro-N-methyl-3-nitrobenzamide (4.99 g, 25.2 mmol) in ethanol (100 mL) was added compound 6 (TFA salt, 10.0 g, 23.97 mmol), followed by DIEA (8.71 mL, 52.7 mmol). The mixture was heated at 85° C. for 6 hours and then at room temperature overnight. The reaction mixture was concentrated in vacuo, and the residue was treated with water and ethyl acetate (200 mL). The yellow solid formed was filtered, washed with diisopropyl ether, and dried to give the title compound 7 (8.8 g, 76% yield) as a yellow powder. TIFF0007789565000203.tif34160

[0263] cis 1-3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiophen-2-yl)-1H-benzo[d]imidazole-5-carboxamide (8f) TIFF0007789565000204.tif38128 Compound 7 (241 mg, 0.5 mmol), thiophene-2-carbaldehyde (72.9 mg, 0.650 mmol), and sodium dithionite (0.261 g, 1.500 mmol) were mixed in 1,4-dioxane (4 mL) and HO (2 mL). The reaction mixture was heated at 130 °C for 1 h under microwave irradiation. The reaction mixture was then concentrated in vacuo. Water was added to the residue, and the aqueous phase was extracted with ethyl acetate (2 × 50 mL). The organic phase was washed with water, then dried over NaSO and concentrated in vacuo. The residue was chromatographed on a silic gel column eluted with CHCl / MeOH = 94 / 6 (v / v) to give the title compound 8f as a yellow solid in 29.4% yield (80 mg). TIFF0007789565000205.tif35160

[0264] cis 1-3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiophen-3-yl)-1H-benzo[d]imidazole-5-carboxamide (8g) TIFF0007789565000206.tif37128 The title compound as a yellow solid was prepared in 33.1% yield similarly to the synthesis of 8f. TIFF0007789565000207.tif35159

[0265] cis 1-3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(2-methylthiazol-4-yl)-1H-benzo[d]imidazole-5-carboxamide (8h) TIFF0007789565000208.tif33128To a solution of compound 7 (241 mg, 0.5 mmol) in 1,4-dioxane (10 mL) / water (1 mL) was added 2-methylthiazole-4-carbaldehyde (95 mg, 0.750 mmol) and sodium dithionite (261 mg, 1.500 mmol). The resultant was heated at 85 °C for 24 h and then cooled to room temperature. The reaction was concentrated in vacuo, and the residue was treated with water and ethyl acetate (20 mL). The off-white solid that formed was filtered off, washed with diisopropyl ether, and dried to give the title compound 8h as an off-white powder (160 mg, 57.3% yield). TIFF0007789565000209.tif27159

[0266] cis 1-3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(thiazol-2-yl)-1H-benzo[d]imidazole-5-carboxamide (8i) TIFF0007789565000210.tif38128 The title compound as a cream-colored powder was prepared in 25.7% yield similarly to the synthesis of 8h. TIFF0007789565000211.tif35159

[0267] cis 1-3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(1H-pyrazol-3-yl)-1H-benzo[d]imidazole-5-carboxamide (8j) TIFF0007789565000212.tif37128 The title compound as a yellow solid was prepared in 70.2% yield similarly to the synthesis of 8h. TIFF0007789565000213.tif35160

[0268] cis 1-3-(5-cyanothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (9Aa) The title compound as a white solid was prepared similarly to the synthesis of 8b, using 5-cyanothiophene-2-carboxylic acid instead of 5-bromothiophene-2-carboxylic acid. TIFF0007789565000215.tif42160

[0269] cis 1-(3-(5-chlorothiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (9Ab) The title compound as a white solid was prepared similarly to the synthesis of 8b, using 5-chlorothiophene-2-carboxylic acid instead of 5-bromothiophene-2-carboxylic acid. TIFF0007789565000217.tif44160

[0270] cis 1-(3-(5-methylthiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (9Ac) The title compound as a white solid was prepared similarly to the synthesis of 8b, using 5-methylthiophene-2-carboxylic acid instead of 5-bromothiophene-2-carboxylic acid. TIFF0007789565000219.tif43160

[0271] cis 1-(3-(5-bromo-N-methylthiophene-2-carboxamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (9Ad) To a solution of 4-fluoro-3-nitrobenzonitrile (0.717 g, 4.31 mmol) in ethanol (20 mL) was added cis N-(-3-aminocyclohexyl)-5-bromothiophene-2-carboxamide (TFA salt, 1.5 g, 3.60 mmol), followed by DIEA (1.307 mL, 7.91 mmol). The reaction mixture was heated at 85 °C for 6 hours and then allowed to stand at room temperature overnight. The ethanol was removed in vacuo, and water was added, followed by ethyl acetate (200 mL). The yellow solid that formed was collected and triturated with diisopropyl ether to give compound I (cis-isomer mixture) (1.25 g, 77% yield) as a yellow powder. TIFF0007789565000221.tif28160 Using compound i and picolinaldehyde as starting materials, compound ii (cis-isomer mixture) as a cream-colored solid was prepared in a 43.4% yield in the same manner as in 8h. To a solution of compound II (950 mg, 1.876 mmol) in DMF (20 mL) was added sodium hydride (90 mg, 3.75 mmol), followed by iodomethane (293 mg, 2.064 mmol). The reaction mixture was stirred at room temperature for 4 hours and then concentrated under reduced pressure. The residue was partitioned between water and ethyl acetate (20 mL). The organic layer was washed twice with water, then dried over anhydrous NaSO and concentrated under reduced pressure. The residue was triturated with diisopropyl ether / CHCl, and the resulting precipitate was filtered and dried to give compound III (cis-isomer mixture) as an off-white solid (910 mg, 93% yield). LC-MS (ESI) m / z = 521.9 (M+H). + A mixture of compound III (894 mg, 1.718 mmol) in sulfuric acid (2000 mg, 20.39 mmol) and water (37.1 mg, 2.061 mmol) was stirred at room temperature for 1 h and then diluted with water and ethyl acetate (20 mL). The mixture was neutralized with NaOH (1 N). The organic layer was washed twice with water, then dried over anhydrous NaSO and concentrated under reduced pressure. The residue was purified by chromatography on a silica gel column eluted with CHCl / MeOH (95 / 5, v / v) to give the desired compound 9Ad (cis-isomer mixture) (200 mg, 21.62% yield) as a white solid. TIFF0007789565000223.tif36159

[0272] cis 1,3-(5-bromothiophene-2-sulfonamido)cyclohexyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (9Ae) TIFF0007789565000224.tif38128 The title compound was prepared analogously to 8b in 43.3% yield (13.4 mg) using 5-bromothiophene-2-sulfonyl chloride instead of 5-bromothiophene-2-carboxylic acid. TIFF0007789565000225.tif36160

[0273] cis-2-Bromo-N-(3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)thiazole-5-carboxamide (9Ba) TIFF0007789565000226.tif36128 The title compound was prepared analogously to 8b in 32% yield (63 mg) using 2-bromothiazole-5-carboxylic acid instead of 5-bromothiophene-2-carboxylic acid. TIFF0007789565000227.tif43160

[0274] cis 1-(3-(5-bromo-1-methyl-1H-pyrrole-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (9Bb) TIFF0007789565000228.tif35128 The title compound was prepared analogously to 8b in 30.7% yield as a cream-colored solid using 5-bromo-1-methyl-1H-pyrrole-2-carboxylic acid instead of 5-bromothiophene-2-carboxylic acid. TIFF0007789565000229.tif42160

[0275] cis-1-(3-(4-bromo-1H-pyrrole-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (9Bc) TIFF0007789565000230.tif32128 The title compound was prepared analogously to 8b in 7.4% yield as a white solid using 4-bromo-1H-pyrrole-2-carboxylic acid instead of 5-bromothiophene-2-carboxylic acid. TIFF0007789565000231.tif35160

[0276] cis-3-chloro-N-(3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)isoxazole-5-carboxamide (9Bd) TIFF0007789565000232.tif35128 The title compound was prepared analogously to 8b in 66.4% yield as a white solid using 3-chloroisoxazole-5-carboxylic acid instead of 5-bromothiophene-2-carboxylic acid. TIFF0007789565000233.tif49160

[0277] cis 5-Bromo-N-(3-(2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)thiophene-2-carboxamide (10a) TIFF0007789565000234.tif39128cis The title compound was prepared analogously to 8f in 16.6% yield (40 mg) as a yellow solid using 5-bromo-N-3-((2-nitrophenyl)amino)cyclohexyl)thiophene-2-carboxamide and picolinaldehyde as starting materials. TIFF0007789565000235.tif43160

[0278] cis 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N,N-dimethyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (10b) Using methyl 4-fluoro-3-nitrobenzoate as the starting material (instead of 4-fluoro-N-methyl-3-nitrobenzamide converted from 4-fluoro-3-nitrobenzoic acid), the corresponding methyl ester (compound iv) was prepared in 30% yield (400 mg) in a similar manner to 8b. Saponification of the methyl ester afforded the corresponding acid v as a crude sample. MS (ESI) m / z [M+1] + = 524.9 and 526.9. The crude acid (60 mg, 0.114 mmol) from above was added to a solution of HATU (43.4 mg, 0.114 mmol), DIEA (40 μL, 0.228 mmol), and dimethylamine (HCl salt, 13.97 mg, 0.171 mmol) in DMF (2 mL) and stirred at room temperature for 1 h. The reaction was concentrated in vacuo, and the residue was purified by HPLC to give the title compound 10b (a mixture of two cis-isomers) in 15.6% yield (11.9 mg). TIFF0007789565000237.tif49159

[0279] cis 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-cyclopropyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (10c) TIFF0007789565000238.tif47128 The title compound was prepared analogously to 10b in 14.7% yield (12.7 mg) using cyclopropylamine instead of dimethylamine. TIFF0007789565000239.tif50160

[0280] cis 1-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-isopropyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (10d) TIFF0007789565000240.tif41128 The title compound was prepared analogously to 10b in 15.7% yield (12.2 mg) using isoproylamine instead of dimethylamine. TIFF0007789565000241.tif43160

[0281] 3-((1S,3R)-3-(5-chlorothiophene-2-carboxamido)cyclohexyl)-2-(3-fluoropyridin-2-yl)-N-methyl-3H-imidazo[4,5-c]pyridine-6-carboxamide Step 1: Synthesis of 4-fluoro-N-methyl-3-nitrobenzamide TIFF0007789565000242.tif20128 To a solution of 4-fluoro-3-nitrobenzoic acid (3.0 g, 16.21 mmol, 1.0 equiv.) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.74 g, 19.51 mmol, 1.2 equiv.) cooled in an ice-water bath was added methylamine solution (12.15 mL of a 2.0 M solution in THF, 24.31 mmol, 1.5 equiv.). The reaction was allowed to warm to room temperature and stirred for 18 hours. The solvent was evaporated in vacuo, and the residue was diluted with ethyl acetate (200 mL). The organic layer was washed with saturated sodium bicarbonate solution (2×150 ml), 0.5 N hydrochloric acid solution (150 ml) and brine (150 ml), dried over magnesium sulfate and the solvent evaporated in vacuo to give 4-fluoro-N-methyl-3-nitrobenzamide (2.10 g, 65% yield) as a yellow solid. TIFF0007789565000243.tif19160

[0282] Step 2: Synthesis of benzyl tert-butyl-cis-cyclohexane-1,3-diyl)dicarbamate TIFF0007789565000244.tif30128 To a solution of Boc-cis-3-aminocyclohexanecarboxylic acid (8.77 g, 36.05 mmol, 1.0 equiv.) and triethylamine (5.53 mL, 39.65 mmol, 1.1 equiv.) in toluene (150 mL) was added diphenylphosphoryl azide (7.79 mL, 36.05 mmol, 1.0 equiv.). The reaction mixture was heated to reflux for 3 hours and then cooled to 80°C. Benzyl alcohol (4.69 mL, 45.06 mmol, 1.25 equiv.) and triethylamine (5.53 mL, 39.65 mmol, 1.1 equiv.) were added, and the reaction was stirred at 80°C for 20 hours. The reaction was cooled to room temperature and diluted with water (100 mL). The aqueous layer was extracted with ethyl acetate (3 x 100 ml), and the combined organic layers were washed with brine (100 ml), dried over magnesium sulfate, and the solvent was evaporated in vacuo. Trituration of the crude product with hexane gave benzyl tert-butyl-cis-cyclohexane-1,3-diyl)dicarbamate (7.81 g, 62% yield) as a colorless solid. TIFF0007789565000245.tif20165

[0283] Step 3: Synthesis of tert-butyl-cis-3-aminocyclohexyl)carbamate To a suspension of palladium on carbon (10 wt.%, 1.19 g, 1.12 mmol, 0.05 equiv.) in ethanol (50 mL) in a Parr hydrogenation flask was slowly added tert-butyl N-[cis-3-(benzyloxycarbonylamino)cyclohexyl]carbamate (2) (7.81 g, 22.4 mmol, 1.0 equiv.) in ethanol (200 mL). The reaction mixture was hydrogenated under 30 psi hydrogen pressure for 3 hours. The reaction mixture was filtered through Celite, and the solvent was evaporated in vacuo to give tert-butyl-cis-3-aminocyclohexyl)carbamate (4.70 g, 97% yield) as a colorless solid. TIFF0007789565000247.tif12145

[0284] Step 4: Synthesis of tert-butyl-cis-3-(((4-(methylcarbamoyl)-2-nitrophenyl)amino)cyclohexyl)carbamate TIFF0007789565000248.tif33128 To a solution of tert-butyl N-[cis-3-aminocyclohexyl]carbamate (2.27 g, 10.59 mmol, 1.0 equiv.) and N,N-diisopropylethylamine (2.28 mL, 12.71 mmol, 1.2 equiv.) in ethanol (30 mL) was added 4-fluoro-N-methyl-3-nitro-benzamide (2.10 g, 10.59 mmol, 1.0 equiv.) in ethanol (20 mL). The reaction was refluxed for 20 hours. The solvent was evaporated in vacuo, and the residue was diluted with water (250 mL). The aqueous layer was extracted with ethyl acetate (4 x 250 ml), dried over magnesium sulfate, and the solvent was evaporated in vacuo to give tert-butyl-cis-3-(((4-(methylcarbamoyl)-2-nitrophenyl)amino)cyclohexyl)carbamate (4.24 g, 100% yield) as a yellow solid. TIFF0007789565000249.tif34160

[0285] Step 5: Synthesis of tert-butyl-(cis-3-((2-amino-4-(methylcarbamoyl)phenyl)amino)cyclohexyl)carbamate To a suspension of palladium on carbon (10 wt.%, 1.15 g, 10.8 mmol, 1.0 equiv.) in ethanol (50 mL) in a Parr hydrogenation flask was slowly added tert-butyl-N-[cis-3-(benzyloxycarbonylamino)cyclohexyl]carbamate (7.81 g, 10.8 mmol, 1.0 equiv.) in ethanol (200 mL). The reaction mixture was hydrogenated under 40 psi hydrogen pressure for 1 hour. The reaction mixture was filtered through Celite, and the solvent was evaporated in vacuo to give tert-butyl-(cis-3-((2-amino-4-(methylcarbamoyl)phenyl)amino)cyclohexyl)carbamate (3.92 g, 100% yield) as a colorless solid. TIFF0007789565000251.tif34160

[0286] Step 6: Synthesis of tert-utyl (cis-3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)carbamate TIFF0007789565000252.tif35128 To a solution of tert-butyl-N-[cis-3-[2-amino-4-(methylcarbamoyl)anilino]cyclohexyl]carbamate (3.92 g, 10.8 mmol, 1.0 equiv.) and 2-pyridinecarboxaldehyde (1.24 mL, 12.96 mmol, 1.2 equiv.) in N,N-dimethylformamide (70 mL) / water (3 mL) was added potassium peroxomonosulfate (4.32 g, 7.02 mmol, 0.65 equiv.). The reaction was stirred for 3 hours. Water (30 mL) was added to the reaction mixture, the pH was adjusted to 9 with 1N sodium hydroxide solution, and the mixture was stirred for 30 minutes. The mixture was extracted with ethyl acetate (4 x 100 ml), washed with brine (150 ml), dried over magnesium sulfate, and the solvent evaporated in vacuo. The crude material was purified by silica chromatography eluting with 0 to 5% methanol in dichloromethane to give tert-butyl-(cis-3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)carbamate (4.46 g, 92% yield) as a colorless solid. TIFF0007789565000253.tif34159

[0287] Step 7: Synthesis of 1-(cis-3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide TIFF0007789565000254.tif40128 To a solution of tert-butyl-(cis-3-(5-(methylcarbamoyl)-2-(pyridin-2-yl)-1H-benzo[d]imidazol-1-yl)cyclohexyl)carbamate (6) (4.46 g, 9.92 mmol, 1.0 equiv.) in dichloromethane (80 mL) was added trifluoroacetic acid (7.59 mL, 99.21 mmol, 10.0 equiv.). The reaction was stirred for 2 hours. The solvent was evaporated in vacuo to give N-methyl-2-(2-pyridyl)-1-[cis-3-aminocyclohexyl]benzimidazole-5-carboxamide trifluoroacetate, which was used directly. 5-Bromothiophene-2-carboxylic acid (3.08 g, 14.88 mmol, 1.5 equiv) in N,N-dimethylformamide (100 ml) was cooled to 0° C. in an ice bath. 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (5.66 g, 14.88 mmol, 1.5 equiv.) and N,N-diisopropylethylamine (10.69 mL, 59.53 mmol, 6.0 equiv.) were added and stirred for 30 minutes. N-Methyl-2-(2-pyridyl)-1-[cis-3-aminocyclohexyl]benzimidazole-5-carboxamide trifluoroacetate (4.60 g, 9.92 mmol) in N,N-dimethylformamide (50 mL) was added. The reaction was allowed to warm to room temperature and stirred for 20 hours. The mixture was stirred. The reaction mixture was poured into saturated sodium bicarbonate solution (200 ml). The mixture was extracted with ethyl acetate (4 x 100 ml), washed with brine (200 ml), dried over magnesium sulfate, and the solvent was evaporated in vacuo. The crude material was purified by silica chromatography eluting with 0 to 5% methanol in dichloromethane to give 1-(cis-3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (3.68 g, 69% yield) as an off-white solid. TIFF0007789565000255.tif34160

[0288] 1-(5-(5-bromothiophene-2-carboxamide)tetrahydro-2H-pyran-3-yl)-6-fluoro-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (234) Preparation of 5-ethoxy-2H-pyran-3(6H)-one TIFF0007789565000256.tif14128 In a 1000 mL round-bottom flask was placed oxane-3,5-dione (10.00 g, 87.64 mmol, 1.00 equiv) in 300 mL of ethanol and 0.28 mL of concentrated HSO. This was stirred at room temperature for 2 hours. The desired product could be detected by LC-MS. The crude product was used directly in the next step without further purification.

[0289] Preparation of 5-amino-2H-pyran-3(6H)-one 5-Ethoxy-2,6-dihydropyran-3-one (approximately 9.20 g, 64.72 mmol, 1.00 equiv.) in 300 mL of ethanol was placed in a 1000 mL round-bottom flask, and NH gas was slowly introduced at 0°C for 3 hours. The desired product could be detected by LC-MS ([M+H] + It was concentrated and the residue was purified on a silica gel column using 40% ethyl acetate in petroleum ether to give 5-amino-2,6-dihydropyran-3-one (6.6 g, 90.2%) as a white solid.

[0290] Preparation of 5-aminotetrahydro-2H-pyran-3-ol TIFF0007789565000258.tif16128In a 500 ml pressure vessel, 5-amino-2,6-dihydropyran-3-one (6.60 g, 58.35 mmol, 1.00 equiv.) in 250 mL of ethanol and Raney Ni (8.00 g) were placed. The mixture was purged with nitrogen three times and then stirred with H2 gas at 100 degrees Celsius under 50 atm. for 2 days. The desired product could be detected by LCMS. The reaction mixture was cooled to room temperature. After filtration, the filtrate was concentrated under vacuum to give 5-aminooxan-3-ol (5.1 g, 74.6%) as a colorless oil. LC-MS: ([M+H] + ):118.1.

[0291] Preparation of benzyl 5-hydroxytetrahydro-2H-pyran-3-carboxylate TIFF0007789565000259.tif18128 In a 100 mL round-bottom flask, 5-aminooxan-3-ol (4.00 g, 34.15 mmol, 1.00 equiv.), benzyl carbonochloridate (6.99 g, 40.97 mmol, 1.20 equiv.), and triethylamine (10.37 g, 102.44 mmol, 3.00 equiv.) in 40 mL of tetrahydrofuran were placed and stirred at room temperature for 4 hours. The desired product could be detected by LC-MS. The solution was concentrated under reduced pressure. After concentration, the residue was purified on a silica gel column using 40% ethyl acetate in petroleum ether to give benzyl N-(5-hydroxyoxan-3-yl)carbamate (3.8 g, 44.3%) as a colorless oil. LC-MS: ([M+H] + ):252.1.

[0292] Preparation of benzyl 5-oxotetrahydro-2H-pyran-3-carboxylate In a 100 mL round-bottom flask, benzyl N-(5-hydroxyoxan-3-yl)carbamate (3.80 g, 15.12 mmol, 1.00 equiv.) and Dess-Martin periodinane (19.24 g, 45.37 mmol, 3.00 equiv.) were placed in 40 mL of dichloromethane. The reaction mixture was stirred at room temperature for 2 h, and the desired product was detected by LCMS. The residue was diluted with 100 mL of NaHCO3 solution and extracted with 3 × 100 mL of dichloromethane. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified on a silica gel column using 50% ethyl acetate in petroleum ether to give benzyl N-(5-oxoxan-3-yl)carbamate (1.9 g, 50.4%) as a colorless oil. LC-MS: ([M+H] + ):250.1.

[0293] Preparation of benzyl 5-oxotetrahydro-2H-pyran-3-carboxylate TIFF0007789565000261.tif21128 In a 100 mL round-bottom flask was placed benzyl N-(5-oxoxooxan-3-yl)carbamate (1.80 g, 7.22 mmol, 1.00 equiv.), tert-butyl carbamate (2.54 g, 21.66 mmol, 3.00 equiv.), triethylsilane (2.519 g, 21.66 mmol, 3.00 equiv.), and 2,2,2-trifluoroacetic acid (1.647 g, 14.44 mmol, 2.00 equiv.) in 50 mL of acetonitrile. The reaction mixture was stirred under N2 at 38°C for 2 days, and the desired product could be detected by LC-MS. The solids were removed by filtration, and the filtrate was concentrated under reduced pressure. The mixture was concentrated and purified by reverse-phase chromatography eluting with 70% acetonitrile in water (0.1% NH4HCO3) to give benzyl N-[5-[(tert-butoxycarbonyl)amino]oxan-3-yl]carbamate (210 mg, 8.3%) as a white solid. The chiral HPLC data indicates that the structure is a cis-trans mixture in a 1:1 ratio. LC-MS: ([M+H] + ):368.2.

[0294] Preparation of benzyl 5-oxotetrahydro-2H-pyran-3-carboxylate TIFF0007789565000262.tif20128 In a 50 mL round-bottom flask, benzyl N-[5-[(tert-butoxycarbonyl)amino]oxan-3-yl]carbamate (160.0 mg, 0.46 mmol, 1.00 equiv.) and Pd / C (9.72 mg, 0.09 mmol, 0.20 equiv.) in 15 mL of isopropanol were stirred under a H atmosphere at room temperature for 4 h. After filtration, the filtrate was concentrated in vacuo to give tert-butyl N-(5-aminooxan-3-yl)carbamate (59 mg, 59.7%) as a white solid. TIFF0007789565000263.tif19159

[0295] Preparation of 1-(5-(5-bromothiophene-2-carboxamido)tetrahydro-2H-pyran-3-yl)-6-fluoro-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide 1-(5-(5-Bromothiophene-2-carboxamide)tetrahydro-2H-pyran-3-yl)-6-fluoro-N-methyl-2-(pyridin-2-yl)-1H-benzo[d]imidazole-5-carboxamide (243) can be prepared from benzyl 5-oxotetrahydro-2H-pyran-3-carboxylate in a manner similar to that described for compounds 286 and 287.

[0296] 3-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-phenyl-1H-indole-6-carboxamide (283) Preparation of benzyl (3-(6-(methylcarbamoyl)-1H-indol-3-yl)cyclohexyl)carbamate TIFF0007789565000264.tif39165 Benzyl (3-oxocyclohexyl)carbamate (1030.38 mg, 4.17 mmol) was added to a solution of N-methyl-1H-indole-6-carboxamide (500 mg, 2.87 mmol) in DCM (10 mL). The solution was cooled to 10 °C, and then trifluoroacetic acid (0.66 mL, 8.61 mmol) was added dropwise, followed by triethylsilane (1.83 mL, 11.48 mmol). The reaction mixture was allowed to warm to room temperature and maintained under stirring. The mixture was poured into a saturated solution of NaHCO3 and extracted three times with ethyl acetate (AcOEt). The combined organic phase was washed with brine, dried over MgSO4, and evaporated. The crude was purified on silica gel using AcOEt / hexane 50 / 50 as the eluent to give 1 g of the desired product (86%). MS: 406.34 [M+H] + .

[0297] Preparation of benzyl (3-(2-bromo-6-(methylcarbamoyl)-1H-indol-3-yl)cyclohexyl)carbamate and benzyl (3-(2-bromo-1-hydroxy-6-(methylcarbamoyl)-1H-indol-3-yl)cyclohexyl)carbamate Pyridinium tribromide (1025.34 mg, 3.21 mmol) was added in one portion to a cooled (ice bath) solution of benzyl N-[3-[6-(methylcarbamoyl)-1H-indol-3-yl]cyclohexyl]carbamate (1000 mg, 2.47 mmol) in THF (5 mL) / chloroform (5 mL). The bath was removed, and the reaction was maintained at room temperature for 2 h. The mixture was washed with 10 mL of a 1 M solution of NaHSO3, 10 mL of 1 N HCl, water, dried over MgSO4, and evaporated. UPLC showed the expected product along with a by-product of MW 500, presumably an oxidized derivative of the expected compound. The two compounds were purified by HPLC using acetonitrile / water (0.1% formic acid) as eluent to give: 390 mg of benzyl (3-(2-bromo-6-(methylcarbamoyl)-1H-indol-3-yl)cyclohexyl)carbamate (33%) and 209 mg of benzyl (3-(2-bromo-1-hydroxy-6-(methylcarbamoyl)-1H-indol-3-yl)cyclohexyl)carbamate (17%).

[0298] Preparation of benzyl (3-(6-(methylcarbamoyl)-2-phenyl-1H-indol-3-yl)cyclohexyl)carbamate To a solution of benzyl N-[3-[2-bromo-6-(methylcarbamoyl)-1H-indol-3-yl]cyclohexyl]carbamate (145 mg, 0.3000 mmol), phenylboronic acid (43.8 mg, 0.3600 mmol), and tetrakis(triphenylphosphine)palladium(0) (34.59 mg, 0.0300 mmol) in DMF (1 mL) was added potassium carbonate (124.12 mg, 0.9000 mmol) in water (0.1000 mL). The mixture was degassed and placed under argon. The reaction was allowed to proceed at 80 °C overnight, and the mixture was poured into a saturated solution of sodium chloride and extracted three times with ethyl acetate. The combined organic phases were washed with water, dried over MgSO4, and evaporated. The crude material was purified on silica gel using CH3OH / CH2Cl2 0-5% as eluent to give 162 mg of the desired product (112%). The product was impure but was used directly in the next step. MS: 482.41 [M+H]+.

[0299] Preparation of 3-(3-aminocyclohexyl)-N-methyl-2-phenyl-1H-indole-6-carboxamide TIFF0007789565000267.tif36148 To benzyl N-[3-[6-(methylcarbamoyl)-2-phenyl-1H-indol-3-yl]cyclohexyl]carbamate (46 mg, 0.1000 mmol) in methanol (10 mL) was added palladium (16.94 mg, 0.1000 mmol). The mixture was hydrogenated in a Parr apparatus under 50 psi H2 atmosphere at room temperature for 2 hours. The mixture was filtered over MgSO4 and evaporated to give 37 mg of a white solid (111%) (UPLC: two peaks of the same mass in 90 / 10). MS: 348.38.

[0300] Preparation of 3-(3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-phenyl-1H-indole-6-carboxamide (283) TIFF0007789565000268.tif41145 3-(3-Aminocyclohexyl)-N-methyl-2-phenyl-1H-indole-6-carboxamide (37 mg, 0.1100 mmol), 5-bromothiophene-2-carboxylic acid (33.07 mg, 0.1600 mmol), HATU (80.98 mg, 0.2100 mmol), N,N-diisopropylethylamine (0.06 mL, 0.3200 mmol), and DMF (2 mL) were combined and stirred at room temperature for 5 h. The mixture was poured into a saturated solution of sodium bicarbonate and extracted three times with ethyl acetate. The combined organic phases were washed with water, dried over MgSO4, and evaporated. The crude material was purified on HPLC using acetonitrile / water (0.1% formic acid) from 20 to 100% to give 30 mg of the desired product (53%). MS:538.28[M+H]+.

[0301] Compounds 290 and 304 were prepared in a similar manner to compound 283.

[0302] 3-((1S,3R)-3-(5-chlorothiophene-2-carboxamido)cyclohexyl)-2-(3-fluoropyridin-2-yl)-N-methyl-3H-imidazo[4,5-c]pyridine-6-carboxamide (286) Preparation of tert-butyl ((1R,3S)-3-(6-chloro-2-(3-fluoropyridin-2-yl)-3H-imidazo[4,5-c]pyridin-3-yl)cyclohexyl)carbamate A mixture of tert-butyl N-[rac-(1R,3S)-3-[(3-amino-2-chloro-4-pyridyl)amino]cyclohexyl]carbamate (507 mg, 1.49 mmol), 3-fluoro-2-formylpyridine (186.08 mg, 1.49 mmol), and oxone (594.38 mg, 0.97 mmol) in DMF / water (10 ml / 1 ml) was heated at 60° C. for 16 hours. The reaction mixture was cooled to room temperature, then diluted with ethyl acetate (30 ml) and washed with water (10 ml) and brine (10 ml). The organic layer was dried over sodium sulfate and evaporated in vacuo to give the crude product. The crude product was purified by silica gel chromatography (eluent, 0 to 100% ethyl acetate / hexanes) to give tert-butyl ((1R,3S)-3-(6-chloro-2-(3-fluoropyridin-2-yl)-3H-imidazo[4,5-c]pyridin-3-yl)cyclohexyl)carbamate (529 mg, 79.7%).

[0303] Preparation of tert-butyl((1R,3S)-3-(6-cyano-2-(3-fluoropyridin-2-yl)-3H-imidazo[4,5-c]pyridin-3-yl)cyclohexyl)carbamate A mixture of tert-butyl N-[rac-(1R,3S)-3-[6-chloro-2-(3-fluoro-2-pyridyl)imidazo[4,5-c]pyridin-3-yl]cyclohexyl]carbamate (529 mg, 1.19 mmol), zinc cyanide (696.42 mg, 5.93 mmol), allylpalladium chloride dimer (43.41 mg, 0.12 mmol), and 2-dicyclohexylphosphinno-2,4,6-triisopropylbiphenyl (XPhos) (113.11 mg, 0.24 mmol) in DMA (7 mL) was heated at 150° C. for 200 minutes under microwave irradiation. The reaction solution was cooled to room temperature, diluted with diethyl acetate (50 mL), and washed with saturated sodium bicarbonate (50 mL). The aqueous solution was extracted with ethyl acetate (20 mL × 2). The combined organic layers were washed with brine (30 mL), dried over sodium sulfate, filtered, and evaporated in vacuo to give the crude product (containing DMA). This crude product was purified on silica gel (12 g, eluent: 0 to 100% ethyl acetate in hexane) to give the desired product, tert-butyl N-[rac-(1R,3S)-3-[6-cyano-2-(3-fluoro-2-pyridyl)imidazo[4,5-c]pyridin-3-yl]cyclohexyl]carbamate (190 mg, 0.4353 mmol, 36.7% yield).

[0304] Preparation of 5-chloro-N-((1R,3S)-3-(6-cyano-2-(3-fluoropyridin-2-yl)-3H-imidazo[4,5-c]pyridin-3-yl)cyclohexyl)thiophene-2-carboxamide A mixture of tert-butyl N-[rac-(1R,3S)-3-[6-cyano-2-(3-fluoro-2-pyridyl)imidazo[4,5-c]pyridin-3-yl]cyclohexyl]carbamate (190 mg, 0.44 mmol) and 2,2,2-trifluoroacetic acid (1 mL, 13.07 mmol) in DCM (3 mL) was shaken at room temperature for 1 h. The reaction solvent was removed by evaporation under reduced pressure and high vacuum pump. The crude product 2-(3-fluoro-2-pyridyl)-3-[rac-(1S,3R)-3-aminocyclohexyl]imidazo[4,5-c]pyridine-6-carbonitrile (140 mg, 0.4162 mmol, 95.6% yield) was used in the next step without further purification.

[0305] A mixture of 2-(3-fluoro-2-pyridyl)-3-[rac-(1S,3R)-3-aminocyclohexyl]imidazo[4,5-c]pyridine-6-carbonitrile (140 mg, 0.4200 mmol), 5-chlorothiophene-2-carboxylic acid (81.21 mg, 0.5 mmol), HATU (237.38 mg, 0.62 mmol), and DIEA (0.23 mL, 1.25 mmol) in DMF (2 mL) was stirred at room temperature for 4 hours. The reaction solution was diluted with ethyl acetate (10 mL) and washed with saturated sodium bicarbonate (5 mL). The aqueous solution was extracted with ethyl acetate (10 mL). The combined organic layers were extracted with brine (5 mL), dried over sodium sulfate, filtered, and evaporated in vacuo to give the crude product. The crude product was purified by silica gel chromatography (12 g, 0–100% EA / Hex) to give the desired product, 5-chloro-N-[rac-(1R,3S)-3-[6-cyano-2-(3-fluoro-2-pyridyl)imidazo[4,5-c]pyridin-3-yl]cyclohexyl]thiophene-2-carboxamide (126 mg, 0.2620 mmol, 62.9% yield).

[0306] Preparation of 3-((1S,3R)-3-(5-chlorothiophene-2-carboxamido)cyclohexyl)-2-(3-fluoropyridin-2-yl)-3H-imidazo[4,5-c]pyridine-6-carboxylic acid A solution of 5-chloro-N-[rac-(1R,3S)-3-[6-cyano-2-(3-fluoro-2-pyridyl)imidazo[4,5-c]pyridin-3-yl]cyclohexyl]thiophene-2-carboxamide (126 mg, 0.26 mmol) in TIFF0007789565000272.tif3812812N HCl (3 ml) was heated at 80° C. for 4 hours. The reaction solution was cooled to room temperature, and excess potassium carbonate was slowly added to neutralize the solution. Ethyl acetate was then added and the solution was extracted several times. The organic layer was dried over sodium sulfate and evaporated in vacuo to give the desired product 2-(3-fluoro-2-pyridyl)-3-[rac-(1S,3R)-3-[(5-chlorothiophene-2-carbonyl)amino]cyclohexyl]imidazo[4,5-c]pyridine-6-carboxylic acid (7, 85 mg, 0.17 mmol, 64.9% yield) without further purification.

[0307] Preparation of 3-((1S,3R)-3-(5-chlorothiophene-2-carboxamido)cyclohexyl)-2-(3-fluoropyridin-2-yl)-N-methyl-3H-imidazo[4,5-c]pyridine-6-carboxamide (286) A mixture of 2-(3-fluoro-2-pyridyl)-3-[rac-(1S,3R)-3-[(5-chlorothiophene-2-carbonyl)amino]cyclohexyl]imidazo[4,5-c]pyridine-6-carboxylic acid (85 mg, 0.17 mmol), HATU (96.97 mg, 0.2600 mmol), DIEA (0.05 mL, 0.2600 mmol), and methylamine (0.17 mL, 0.34 mmol, 2.0 M in THF) in DMF (2 ml) was stirred at room temperature for 3 hours. The crude product was purified by prep-HPLC (30–100% ACN) to give the desired product, 2-(3-fluoro-2-pyridyl)-N-methyl-3-[rac-(1S,3R)-3-[(5-chlorothiophene-2-carbonyl)amino]cyclohexyl]imidazo[4,5-c]pyridine-6-carboxamide 286 (41 mg, 0.0799 mmol, 47% yield).

[0308] 3-((1R,3S)-3-(5-bromothiophene-2-carboxamido)cyclohexyl)-N-methyl-2-(pyridin-2-yl)-3H-imidazo[4,5-c]pyridine-6-carboxamide (287) Step 1: Synthesis of Compound 287a TIFF0007789565000274.tif33128 In a 250 mL three-necked round-bottom flask were placed cis-3-[(tert-butoxycarbonyl)amino]cyclohexanecarboxylic acid (10.0 g, 41.1 mmol), triethylamine (9.0 mL, 64.50 mmol), and diphenylphosphoryl azide (10.0 mL, 46.35 mmol) in 250 mL of toluene, and the resulting mixture was stirred at room temperature for 3 hours. To this was added benzyl alcohol (8.50 mL, 82.00 mmol), and the mixture was stirred at 100° C. for 4 hours. The desired product was detected by LCMS, and the starting acid had been consumed. The reaction mixture was cooled and then concentrated, and the residue was purified on silica gel using 30% ethyl acetate in petroleum ether to give tert-butylbenzyl ((1R,3S)-cyclohexane-1,3-diyl)dicarbamate (287a) (11.2 g, 78.2%) as a white solid. LC-MS m / z: [M-100+H] + :249.2.

[0309] Step 2: Synthesis of Compound 287b TIFF0007789565000275.tif321282 A 50 mL round-bottom flask containing tert-butylbenzyl ((1R,3S)-cyclohexane-1,3-diyl)dicarbamate (8.0 g, 233.78 mmol, 1.00 equiv.) and Pd / C (800.0 mg) in 200 mL of ethyl acetate was stirred under H2 atmosphere at room temperature overnight and monitored by TLC. After filtration, the filtrate was concentrated in vacuo to give tert-butyl ((1S,3R)-3-aminocyclohexyl)carbamate (287b) (4.45 g, 90.4%) as a light yellow oil.

[0310] Step 3: Synthesis of Compound 287c TIFF0007789565000276.tif26128 In a 500 mL three-necked round-bottom flask was placed 2-chloro-5-fluoropyridine (10.00 g, 76.00 mmol, 1.00 equiv.) in 200 mL of 2,2,2-trifluoroacetic acid and 25 mL of 50% hydrogen peroxide. The resulting mixture was stirred at 70 °C until TLC indicated consumption of the starting material. The reaction mixture was concentrated in vacuo and coevaporated with 2 × 40 mL of toluene. The residue was diluted with 40 mL of water and 200 mL of dichloromethane, and the pH of the solution was adjusted to 8 with 28% ammonium hydroxide solution. The aqueous layer was extracted with 3×200 mL of dichloromethane, and the combined organic layers were dried over anhydrous NaSO, filtered, concentrated, and the crude product was purified on a silica gel column using 10% methanol in dichloromethane to give 2-chloro-5-fluoropyridine-1-oxide (287c) (8.10 g, 71.2%) as a yellow solid.

[0311] Step 4: Synthesis of compound 287d TIFF0007789565000277.tif30128 Into a 500 mL three-necked round-bottom flask was placed 2-chloro-5-fluoropyridine 1-oxide (8.10 g, 54.90 mmol, 1.00 equiv.) in 110 mL of concentrated H2SO4. To this was then added 23.90 g of potassium nitrate in portions at 0 °C. The reaction mixture was stirred at 120 °C for 2 hours and monitored by TLC, which showed that the starting material had been consumed. The reaction mixture was then quenched with 400 mL of water / ice. The pH value of the solution was adjusted to 8 with 28% ammonium hydroxide solution while maintaining the temperature below 15 °C in an ice bath. The precipitated light yellow crystals were collected by filtration, washed with 2×50 mL of water, and dried to give 2-chloro-5-fluoro-4-nitropyridine 1-oxide (287d) (6.60 g, 62.4%) as a light yellow solid. The crude product was used directly in the next step without further purification.

[0312] Step 5: Synthesis of Compound 287e Into a 100 mL three-necked round-bottom flask was placed 2-chloro-5-fluoro-4-nitropyridine 1-oxide (287d) (6.60 g, 34.28 mmol, 1.00 equiv.) and tert-butyl ((1S,3R)-3-aminocyclohexyl)carbamate (287b) (7.36 g, 34.28 mmol, 1.00 equiv.) in 20 mL of 1-methylpyrrolidin-2-one. The reaction mixture was stirred at 90° C. for 4 hours, the solution was cooled, and then quenched with 100 mL of ice / water. The precipitated yellow crystals were collected by filtration, washed with 2×50 mL of ice / water, and dried to give 5-(((1R,3S)-3-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-chloro-4-nitropyridine 1-oxide (287e) (11.65 g, 87.9%) as a yellow solid. LC-MS (ESI) m / z [M+H] + :387.2, 389.2.

[0313] Step 6: Synthesis of compound 287f TIFF0007789565000279.tif45128 Into a 100 mL three-necked round-bottom flask was placed 5-(((1R,3S)-3-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-chloro-4-nitropyridine 1-oxide (287e) (11.60 g, 29.99 mmol, 1.00 equiv), Fe (5.02 g, 89.96 mmol, 3.00 equiv), and NHCl (4.81 g, 89.96 mmol, 3.00 equiv) in 20 mL of ethanol / 20 mL of water. The reaction mixture was stirred at 90 °C under N for 2 h. The solids were removed by filtration, and the filtrate was concentrated under reduced pressure. The mixture was concentrated and purified by reverse-phase chromatography eluting with 65% acetonitrile in water (0.1% formic acid) to give tert-butyl N-[(1S,3R)-3-[(4-amino-6-chloropyridin-3-yl)amino]cyclohexyl]carbamate (287f) (4.45 g, 43.5%) as a light yellow solid. TIFF0007789565000280.tif27160

[0314] Step 7: Synthesis of Compound 287g TIFF0007789565000281.tif47135 In a 50 mL round-bottom flask was added tert-butyl N-[(1S,3R)-3-[(4-amino-6-chloropyridin-3-yl)amino]cyclohexyl]carbamate (2.45 g, 7.19 mmol, 1.00 equiv.), picolinic acid (0.88 g, 7.19 mmol), and 15 mL of N,N-dimethylformamide. To a solution of N-ethyl-N-isopropylpropan-2-amine (2.79 g, 21.56 mmol, 3.00 equiv.) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (3.28 g, 8.63 mmol, 1.20 equiv.) was added at 0°C and stirred at room temperature overnight. The solution was concentrated under reduced pressure and purified by reverse-phase chromatography eluting with 60% acetonitrile in water (0.1% formic acid) to give tert-butyl N-[(1S,3R)-3-[[6-chloro-4-(pyridin-2-amido)pyridin-3-yl]amino]cyclohexyl]carbamate (1.40 g, 43.7%) as a purple solid. LC-MS(ESI)m / z[M+H] + :446.2, 448.2.

[0315] Step 8: Synthesis of Compound 287h TIFF0007789565000282.tif48140A 50 mL round-bottom flask was charged with tert-butyl N-[(1S,3R)-3-[[6-chloro-4-(pyridin-2-amido)pyridin-3-yl]amino]cyclohexyl]carbamate (1.40 g, 3.14 mmol, 1.00 equiv.), 4-methylbenzenesulfonic acid (5.41 g, 31.39 mmol, 10 equiv.) in 12 mL of 1-methylpyrrolidin-2-one and stirred at 130 degrees Celsius overnight. The mixture was concentrated and purified by reverse-phase chromatography eluting with 55% acetonitrile in water (0.1% formic acid) to give (1S,3R)-3-[6-chloro-2-(pyridin-2-yl)imidazo[4,5-c]pyridin-3-yl]cyclohexan-1-amine (287h) (650 mg, 63.2%) as a light yellow solid. LC-MS (ESI) m / z [M+H] + :328.1, 330.1.

[0316] Step 9: Synthesis of Compound 287i A solution of (1S,3R)-3-[6-chloro-2-(pyridin-2-yl)imidazo[4,5-c]pyridin-3-yl]cyclohexane-1-amine (650.0 mg, 1.98 mmol, 1.00 equiv.), di-tert-butyl dicarbonate (519.3 mg, 2.38 mmol, 1.20 equiv.), and N-ethyl-N-isopropylpropan-2-amine (768.8 mg, 5.95 mmol, 3.00 equiv.) in 10 mL of dichloromethane was placed in a 50 mL round-bottom flask and stirred at room temperature for 3 hours. The solution was concentrated in vacuo. The resulting residue was purified by reverse-phase chromatography eluting with 60% acetonitrile in water (0.1% formic acid) to afford tert-butyl ((1S,3R)-3-(6-chloro-2-(pyridin-2-yl)-3H-imidazo[4,5-c]pyridin-3-yl)cyclohexyl)carbamate (287i) (356 mg, 41.9%) as a yellow solid. LC-MS (ESI) m / z [M+H] + :428.1, 430.1.

[0317] Step 10: Synthesis of Compound 287j tert-Butyl N-[(1S,3R)-3-[6-chloro-2-(pyridin-2-yl)imidazo[4,5-c]pyridin-3-yl]cyclohexyl]carbamate (350.00 mg, 0.82 mmol, 1.00 equiv.), Pd(dppf)Cl (777.99 mg, 1.06 mmol, 1.30 equiv.), and trimethylamine (248.29 mg, 2.45 mmol, 3.00 equiv.) were placed in a 30 mL pressure vessel in 8 mL of methanol / 4 mL of (methylsulfinyl)methane. The mixture was purged with nitrogen three times and then pressurized to 30 atm with carbon monoxide at 100°C overnight. The reaction mixture was cooled to room temperature. The mixture was concentrated and purified by reverse-phase chromatography eluting with 58% acetonitrile in water (0.1% formic acid) to give methyl 3-[(1R,3S)-3-[(tert-butoxycarbonyl)amino]cyclohexyl]-2-(pyridin-2-yl)imidazo[4,5-c]pyridine-6-carboxylate (158 mg, 42.8%) as a yellow solid. LC-MS (ESI) m / z [M+H] + :452.2, RT=0.70 minutes.

[0318] Step 11: Synthesis of compound 287k TIFF0007789565000285.tif43128 Into a 50 mL round-bottom flask was placed methyl 3-[(1R,3S)-3-[(tert-butoxycarbonyl)amino]cyclohexyl]-2-(pyridin-2-yl)imidazo[4,5-c]pyridine-6-carboxylate (158.00 mg, 0.35 mmol, 1.00 equiv.) in 20 mL of 4 M HCl (g) in 1,4-dioxane. This was stirred at room temperature for 3 hours. The solution was concentrated to give 120 mg of a yellow solid. This crude product was used directly in the next step without further purification. LC-MS (ESI) m / z [M+H] + :352.2.

[0319] Step 12: Synthesis of Compound 287l TIFF0007789565000286.tif461302 In a 5 mL round-bottom flask, 5-bromothiophene-2-carboxylic acid (120.0 mg, 0.58 mmol, 1.20 equiv.), 3-[(1R,3S)aminocyclohexyl]2(pyridinyl)imidazo[4,5c]pyridine carboxylate methyl (169.7 mg, 0.48 mmol, 1.0 equiv.) in 5 mL of N,N-dimethylformamide were added. 0.00 equiv.), N-ethyl-N-isopropylpropan-2-amine (187.3 mg, 1.45 mmol, 3.00 equiv.), and to this was added O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (220.4 mg, 0.58 mmol, 1.20 equiv.) in 5.00 mL of N,N-dimethylformamide dropwise at 0 degrees Celsius, and this was stirred at room temperature for 4 hours. The mixture was purified by reverse-phase chromatography eluting with 52% acetonitrile in water (0.1% formic acid) to afford methyl 3-[(1R,3S)-3-(5-bromothiophene-2-amido)cyclohexyl]-2-(pyridin-2-yl)imidazo[4,5-c]pyridine-6 ​​carboxylate 287l (103.3 mg, 56.1%) as a white solid. TIFF0007789565000287.tif34159

[0320] Step 13: Synthesis of compound 287m TIFF0007789565000288.tif43131 A 50 mL round-bottom flask was charged with methyl 3-[(1R,3S)-3-(5-bromothiophene-2-amido)cyclohexyl]-2-(pyridin-2-yl)imidazo[4,5-c]pyridine-6-carboxylate (90.0 mg, 0.17 mmol, 1.00 equiv.), LiOH.HO (34.9 mg, 0.84 mmol, 5.00 equiv.) in 5 mL of methanol / 5 mL of water and stirred at room temperature for 2 hours. The mixture was concentrated and purified by reverse-phase chromatography eluting with 60% acetonitrile in water (0.1% formic acid) to give 3-[(1R,3S)-3-(5-bromothiophene-2-amido)cyclohexyl]-2-(pyridin-2-yl)imidazo[4,5-c]pyridine-6-carboxylic acid 287m (59.4 mg, 67.8%) as a white solid. TIFF0007789565000289.tif41160

[0321] Step 14: Synthesis of Compound 287 TIFF0007789565000290.tif47145In a 50 mL round-bottom flask was added 3-[(1R,3S)-3-(5-bromothiophene-2-amido)cyclohexyl]-2-(pyridin-2-yl)imidazo[4,5-c]pyridine-6-carboxylic acid (55.0 mg, 0.11 mmol, 1.00 equiv.) in 3 mL of N,N-dimethylformamide. Cetylamine hydrochloride (6.9 mg, 0.095 mmol, 1.00 equiv.), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (43.3 mg, 0.11 mmol, 1.2 equiv.), and N-ethyl-N-isopropylpropan-2-amine (36.8 mg, 0.29 mmol, 3.00 equiv.) were added. The reaction mixture was stirred at room temperature for 2 h and purified by reverse-phase chromatography eluting with 60% acetonitrile in water (0.1% formic acid) to give methyl 3-[(1R,3S)-3-(5-bromothiophene-2-amido)cyclohexyl]-N-methyl-2-(pyridin-2-yl)imidazo[4,5-c]pyridine-6-carboxamide 287 (53.2 mg, 94.0%) as a white solid. TIFF0007789565000291.tif34159

[0322] Further compounds Additional compounds disclosed herein were prepared in a manner similar to the above procedure. The following table shows mass spectrometry data for selected compounds disclosed herein.

[0323] TIFF0007789565000292.tif238154TIFF0007789565000293.tif238154TIFF00077895650 00294.tif238154TIFF0007789565000295.tif238154TIFF0007789565000296.tif164154

[0324] The following table shows the activity of selected compounds disclosed herein. 1 H-NMR data is shown.

[0325] TIFF0007789565000297.tif40160TIFF0007789565000298.tif225160TIFF0007789565000299.tif225160TIFF0007789565000300.tif225160 TIFF0007789565000301.tif243160TIFF0007789565000302.tif217160TIFF0007789565000303.tif225160TIFF0007789565000304.tif225160 TIFF0007789565000305.tif230160TIFF0007789565000306.tif243160TIFF0007789565000307.tif234160TIFF0007789565000308.tif225160 TIFF0007789565000309.tif243160TIFF0007789565000310.tif230160TIFF0007789565000311.tif239160TIFF0007789565000312.tif180160

[0326] equivalent Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments specifically described herein which equivalents are intended to be encompassed by the scope of the appended claims.

[0327] INCORPORATION BY REFERENCE All patents and publications mentioned herein are incorporated by reference in their entirety.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof: (In the formula, Z is CH 2 , C.F. 2 , O or a bond; J 1 , J 2 , J 3 and J. 4 each is independently CH or N; X 1 is N or CR 5A and; X 2 is N or CR 5B and; R 1 Ha-NR 3 C(O)R 4 , or -C(O)(CH 2 ) m NR 3 R 4 and; R 2 is C 3 ~C 8 Cycloalkyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 , C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S- (haloC 1 ~C 6 alkyl), C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 forming a cycloalkyl; R 3 is H or C 1 ~C 6 is alkyl; R 4 is H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 , C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 Aryl, -(CH 2 ) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with; R 5A is F; R 5B H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 Is it; Or R 5A and R 5B together with the atoms to which they are attached form a 6-membered heterocyclyl ring which may be substituted with oxo; R 6 and R 7 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 is cycloalkyl; Each R 8 are independently H, C 1 ~C 6 Alkyl or C 3 ~C 8 is cycloalkyl; Each R 9 are independently H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl or C 3 ~C 8 is cycloalkyl; m is 0 or 1; q is 0 or 1; r is 0 or 1) A compound or a pharmaceutically acceptable salt thereof.

2. J 2 , J 3 and J. 4 Each of the following is CH; 1 is CR 5A and X 2 is CR 5B 2. The compound of claim 1, wherein:

3. R 4 each containing one, two or three substituents R 4' C optionally substituted with 6～10 10. The compound of claim 1, which is aryl or 5-10 membered heteroaryl.

4. R 4 is selected from phenyl, thiophene, oxazole, isoxazole, thiazole, furan and pyrrole, each of which may contain one, two or three substituents R 4' where each R 4' -F, -Cl, -Br, -CN, -NO 2 , -CF 3 , -CH 3 , -CH(CH 3 ) 2 , -C(O)CH 3 , -C(O)OCH 3 4. The compound of claim 3, wherein each of the aryl groups is independently selected from the group consisting of phenyl, cyclopropyl, and morpholinyl.

5. R 2 each containing one, two or three substituents R 2' C optionally substituted with 6～10 The compound of any one of claims 1 to 4, which is aryl or 5-10 membered heteroaryl.

6. R 2 is selected from the group consisting of phenyl, pyridine, pyrimidine, pyridazine, pyrazine, imidazole or thiophene, each of which may contain one, two or three substituents R 2' where each R 2' -F, -Cl, -Br, -CN, NO 2 , -CH 3 , -CF 2 H, -C≡CH, -C(O)H, -CONH 2 , -C(O)NHCH 3 , -OH, -OCH 3 , -OCF 3 , -SCF 3 , -NH 2 , -NHC(O)CH 3 6. The compound of claim 5, wherein the compound is independently selected from the group consisting of:

7. X 2 is CR 5B and R 5B -C(O)NR 8 R 9 7. The compound of any one of claims 1 to 6, wherein

8. X 2 is CR 5B and R 5B Halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 7. The compound of any one of claims 1 to 6, wherein

9. X 2 is CR 5B and R 5B Ga-NR 8 COR 9 or -C(O)OR 8 9. The compound of claim 8, wherein:

10. X 2 is CR 5B and R 5B Ga-CF 3 , -COCH 3 , -CH 2 7. The compound of any one of claims 1 to 6, which is OH, -CN or tetrazole.

11. 10. The compound of claim 1 having the structure of formula (III) or a pharmaceutically acceptable salt thereof: During the ceremony, each J 3 and J. 4 are independently CH or N; X 1 is N or CR 5A and; X 2 is N or CR 5B and; Here, J 3 , J 4 , X 1 and X 2 of which 1, 2, 3 or 4 are N; R 2 is C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 , C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S- (haloC 1 ~C 6 alkyl), C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 forming a cycloalkyl; R 4 is H, C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 , C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 3 ~C 8 Cycloalkyl, C 6 ~C 10 Aryl, -(CH 2 ) q NR 8 C(O)R 9 , -C(O)NR 8 R 9 , -C(O)OR 9 , -C(O)R 9 and one, two or three substituents R independently selected from 5- to 10-membered heterocyclyl 4' may be substituted with.

12. R 2 But, halo, -CN, -NO 2 and C 1 ~C 6 one, two or three substituents R independently selected from alkyl 2' 12. The compound of claim 11, wherein the phenyl is optionally substituted with

13. R 2 But, halo, -CN, -NO 2 and C 1 ~C 6 one, two or three substituents R independently selected from alkyl 2' 12. The compound of claim 11, wherein the heteroaryl is a 5- to 10-membered heteroaryl optionally substituted with:

14. R 4 14. The compound of any one of claims 11-13, wherein is thiophene optionally substituted with halo or phenyl.

15. X 2 is CR 5B and; R 5B -Hydrogen, -halo, -C(O)NR 8 R 9 , -NR 8 COR 9 or -C(O)OR 8 Selected from; R 8 and R 9 are each independently H or C 1 ~C 6 is alkyl, 15. The compound of any one of claims 11 to 14.

16. 10. The compound of claim 1 having the structure of formula (IV) or a pharmaceutically acceptable salt thereof: During the ceremony, R 1 Ha-NR 3 C(O)R 4 , or -C(O)(CH 2 ) m NR 3 R 4 and; R 2 is C 3 ~C 8 Cycloalkyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 , C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 It forms a cycloalkyl.

17. 10. The compound of claim 1 having the structure of formula (V) or a pharmaceutically acceptable salt thereof: During the ceremony, R 1 Ha-NR 3 C(O)R 4 , or -C(O)(CH 2 ) m NR 3 R 4 and; R 2 is C 3 ~C 8 Cycloalkyl, C 6 ~C 10 aryl or 5-10 membered heteroaryl, where each aryl or heteroaryl is selected from halo, -CN, -NO 2 , C 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, HaloC 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkoxy, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, -C(O)NR 6 R 7 , -C(O)R 6 , -NR 6 R 7 , -NR 6 C(O)R 7 , -OH, -S- (haloC 1 ~C 6 alkyl), C 6 ~C 10 one, two, or three substituents R independently selected from aryl and 5- to 10-membered heterocyclyl; 2' or two R 2' The substituents, together with the atoms to which they are attached, are each optionally substituted with halo, or C 3 ~C 8 forming a cycloalkyl; R 5A is F; R 5B H, halo, -CN, C 1 ~C 6 Alkyl, HaloC 1 ~C 6 Alkyl, C 1 ~C 6 Alkoxy, -(CH 2 ) r OH, -C(O)NR 8 R 9 , -NR 8 COR 9 , 5-10 membered heteroaryl, -C(O)R 8 or -C(O)OR 8 Is it; Or R 5A and R 5B are taken together with the atoms to which they are attached to form a 6-membered heterocyclyl ring which may be substituted with oxo.

18. 10. The compound of claim 1 having the structure of formula (VI) or a pharmaceutically acceptable salt thereof: 。

19. 10. The compound of claim 1 having the structure of formula (VII) or a pharmaceutically acceptable salt thereof: 。

20. below: A compound selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

21. the below described:

1. A compound selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

22. the below described:

1. A compound selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

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