Bicyclic heteroarenes and methods of use thereof

Abstract

Disclosed is a compound useful for treating neurological disorders.The compounds described herein can be used alone or in combination with other pharmacoactive agents to treat or prevent nervous system diseases, including frontotemporal dementia, chronic traumatic encephalopathy, Alzheimer's disease, limbic-predominant age-related TDP-43 encephalopathy (LATE) or frontotemporal lobar degeneration.The present invention relates to bicyclic heteroarenes and their use for therapeutic treatment of neurological disorders in patients, such as human patients.

Description

[Technical field]

[0001] FIELD OF THEINVENTION The present invention relates to bicyclic heteroarenes and their use for the therapeutic treatment of neurological disorders in a patient, such as a human patient. [Background technology]

[0002] background Incomplete understanding of the molecular perturbations that cause disease and limited availability of robust model systems have contributed to the failure to generate successful disease-modifying therapies for common progressive neurological disorders such as ALS and FTD. Progress has been made in many frontier fields to find drugs that can block the progression of these disorders. However, most, if not all, current treatments for these diseases provide only very little relief. Thus, there is a need to develop therapies that can modify the course of neurodegenerative diseases. More generally, there is a need for better methods and compositions for treating neurodegenerative diseases to improve the quality of life of people suffering from such diseases. Summary of the Invention [Means for solving the problem]

[0003] Abstract TDP-43 is a nuclear DNA / RNA binding protein involved in RNA splicing. Under pathological cellular stress, TDP-43 translocates to the cytoplasm and aggregates into stress granules and associated protein inclusions. These phenotypes are hallmarks of degenerating motor neurons and are found in 97% of all ALS cases. The high penetrance of this pathology indicates that TDP-43 is broadly involved in both familial and sporadic ALS. Furthermore, TDP-43 mutations that promote aggregation are linked to a higher risk of developing ALS, suggesting that protein misfolding and aggregation act as drivers of toxicity. TDP-43 toxicity can be recapitulated in a yeast model, where the protein causes viability deficits and localizes to stress granules. In one aspect, the present invention provides a compound of formula (I): [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, [ka] is a single bond, and X 1 is (C(R A )2) m or -OC(R A )2-R X and X 2 is C(R A )2 or CO, or [ka] is a double bond and X 1 and X 2 are each independently A or N, where R X X 2 is a bond to R 1 , -(L) n -R B Halo, cyano, hydrogen, optionally substituted C 1~6Alkoxy, optionally substituted C containing at least one ring oxygen 1~9 Heterocyclyl, optionally substituted C1-C6 alkyl, optionally substituted piperazin-1-yl, optionally substituted pyrrolidin-3-yl, pyrimidinyl (wherein the pyrimidinyl is cyclopropyl or optionally substituted C6-C 10 aryl), optionally substituted pyridazinyl, optionally substituted oxazolyl, pyrid-2-on-1-yl, optionally substituted isoindolinyl, unsubstituted pyridin-4-yl, unsubstituted pyridin-2-yl, optionally substituted furan-3-yl, unsubstituted pyridin-3-yl or optionally substituted pyrazol-1-yl; R 2 is an optionally substituted C1-C6 alkyl, an optionally substituted C6-C 10 Aryl, optionally substituted piperidin-4-yl, optionally substituted tetrahydropyran-4-yl, optionally substituted pyrimidin-5-yl, optionally substituted pyrimidin-4-yl, optionally substituted pyridin-3-yl, optionally substituted pyridazin-4-yl, optionally substituted pyrazol-1-yl, optionally substituted pyrazol-4-yl, optionally substituted pyrazol-3-yl, optionally substituted pyridin-2-yl, optionally substituted triazolyl, optionally substituted benzodioxol-2-yl, optionally substituted benzodioxan-2-yl, optionally substituted C6-C 10 Aryl C1-C 10 is alkyl or optionally substituted acyl; R 3 is a group having the following structure: [ka] and R Aeach independently represents H, optionally substituted C 1~6 Alkyl or optionally substituted C6-C 10 Aryl or two R A together with the atom to which they are attached form an oxo, where two R A When they combine with the atom to which they are attached to form an oxo, [ka] is a single bond, R B is replaced as necessary 6~10 Aryl, optionally substituted C1-C9 heteroaryl, optionally substituted C 3~8 Cycloalkyl, -N=CH-R D or optionally substituted C1-C9 heterocyclyl, optionally substituted C2-C9 heteroaryl C1-C6 alkyl, or optionally substituted C2-C9 heterocyclyl C1-C6 alkyl; R C is H or optionally substituted C1-C6 alkyl; R D is optionally substituted C6-C 10 is aryl, or Each L is independently an optionally substituted C 1~6 Alkylene, optionally substituted C1-C6 heteroalkylene, optionally substituted C3-C8 cycloalkylene, optionally substituted C2-C6 alkynylene, O or NR C and n is 1, 2 or 3; m is 0, 1 or 2).

[0004] In some embodiments, [ka] is a single bond. In some embodiments, X 1is (C(R A )2) m In some embodiments, m is 1. In some embodiments, X 2 is C(R A In some embodiments, R A are each hydrogen.

[0005] In some embodiments, the compound has formula (Ia): [ka] or a pharma- ceutically acceptable salt thereof.

[0006] Preferably, the compound of formula Ia has the following structure: [ka] or a pharma- ceutically acceptable salt thereof.

[0007] In some embodiments, the compound has formula (1a'): [ka] or a pharma- ceutically acceptable salt thereof.

[0008] Preferably, the compound of formula Ia' has the following structure: [ka] or a pharma- ceutically acceptable salt thereof.

[0009] In some embodiments, the compound has formula (1b): [ka] or a pharma- ceutically acceptable salt thereof.

[0010] In some embodiments, the compound has formula (1c): [ka] or a pharma- ceutically acceptable salt thereof.

[0011] In some embodiments, the compound has formula (1d): [ka] or a pharma- ceutically acceptable salt thereof.

[0012] In some embodiments, the compound has formula (1e): [ka] or a pharma- ceutically acceptable salt thereof.

[0013] In some embodiments, R 1 is -O-(L) (n-1) -R B In some embodiments, n is 2. In some embodiments, n is 1. In some embodiments, at least one L is optionally substituted C 1~6 In some embodiments, L is alkylene. In some embodiments, L is methylene. In some embodiments, L is ethylene. In some embodiments, R B is replaced as necessary 1~9 In some embodiments, R B is replaced as necessary 1~9 In some embodiments, R is heteroaryl. B is replaced as necessary 1~6 It is an alkyl.

[0014] In some embodiments, R A is replaced as necessary 1~6 alkyl (e.g., methyl, ethyl, propyl, butyl, penyl, hexyl). In some embodiments, R A is methyl. In some embodiments, R 1 is replaced as necessary1~6 alkyl (e.g., methyl, ethyl, propyl, butyl, penyl, hexyl). In some embodiments, R 1 is an optionally substituted C alkyl. In some embodiments, R 1 is substituted with hydroxyl. In some embodiments, R 1 is optionally substituted piperazin-1-yl. In some embodiments, R 1 is substituted with methyl. In some embodiments, R 2 is optionally substituted C6-C 10 arylC1-C6 alkyl. In some embodiments, R 2 is optionally substituted C6-C 10 arylC1-C2 alkyl. In some embodiments, R 2 is substituted with oxo. 2 is optionally substituted C1-C6 alkyl. In some embodiments, R 2 is an optionally substituted C alkyl. In some embodiments, R 2 is substituted with hydroxyl. In some embodiments, R 2 is optionally substituted C6-C 10 In some embodiments, R 2 is optionally substituted phenyl. In some embodiments, R 2 is substituted with fluoro, cyano or methoxy.

[0015] In some embodiments, R 1 teeth, [ka] [ka] It is methyl or methoxy.

[0016] In some embodiments, R 1teeth, [ka] Or methoxy.

[0017] In some embodiments, R 2 teeth, [ka] It is.

[0018] In some embodiments, R 2 teeth, [ka] It is.

[0019] In some embodiments, R 3 teeth, [ka] It is.

[0020] In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 2 is optionally substituted pyrimidin-3-yl or optionally substituted pyrimidin-4-yl; R 4 is hydrogen or optionally substituted C6-C 10 (aryl).

[0021] In some embodiments, R 4 is hydrogen. In some embodiments, R 4 is optionally substituted C6-C 10 In some embodiments, R 4 C6~C 10 Aryl is phenyl. In some embodiments, R4 is substituted with fluoro. 4 is substituted with methoxy. 4 is optionally substituted pyridin-3-yl. In some embodiments, R 4 is pyridin-4-yl.

[0022] In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 5 is hydrogen or optionally substituted C6-C 10 is aryl, R 2 is an optionally substituted triazolyl, an optionally substituted pyrazol-4-yl, an optionally substituted pyrazol-3-yl, an optionally substituted pyrimidin-4-yl or an optionally substituted C6-C 10 aryl C1-C6 alkyl).

[0023] In some embodiments, R 5 is hydrogen. In some embodiments, R 5 is phenyl. In some embodiments, R 2 is substituted with phenyl. In some embodiments, R 2 teeth, [ka] In some embodiments, R 2 teeth, [ka] It is.

[0024] In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 2 is optionally substituted pyridin-3-yl). In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 2 is optionally substituted C6-C 10 aryl; or optionally substituted pyridazin-4-yl).

[0025] In some embodiments, R 2 is 3-fluoro-phenyl. In some embodiments, R 2 is pyridazin-4-yl.

[0026] In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 2 is optionally substituted pyridin-3-yl).

[0027] In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof. (wherein L is an optionally substituted C3-C8 cycloalkylene or C2-C6 alkynylene; R B is optionally substituted C6-C 10 (aryl).

[0028] In some embodiments, L is [ka] In some embodiments, L is [ka] In some embodiments, R B is 3-methoxy-phenyl.

[0029] In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, LR B is -NHN=CHR D and R 2 is optionally substituted pyridin-3-yl; R D is optionally substituted C6-C 10 (aryl).

[0030] In some embodiments, R D is 3-methyl-phenyl. In some embodiments, R 2 is pyridin-3-yl. In some embodiments, the compound has the structure [ka] or a pharma- ceutically acceptable salt thereof.

[0031] In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 2 is optionally substituted pyridin-3-yl; R 6 is optionally substituted C6-C 10 (aryl).

[0032] In some embodiments, R 7 is 3-methyl-phenyl or phenyl.

[0033] In one aspect, the present invention provides a compound of formula (12) [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, X 3 is N or CH, R 8 is an optionally substituted C2-C9 heteroaryl; or an optionally substituted C6-C 10 is aryl, R 9 is an optionally substituted C2-C9 heteroaryl or -OR 11 and R 10 is hydrogen or optionally substituted C1-C6 alkyl; R 11 is optionally substituted C2-C9 heteroaryl C1-C6 alkyl).

[0034] In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof.

[0035] In some embodiments, R 2 is 2-hydroxy-ethyl. In some embodiments, R 2 is hydrogen.

[0036] In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof.

[0037] In some embodiments, the compound has the structure: [ka] or a pharma- ceutically acceptable salt thereof.

[0038] In some embodiments, R A is optionally substituted phenyl. In some embodiments, R A is optionally substituted pyridin-2-yl, optionally substituted pyrimidin-4-yl, optionally substituted pyrimidin-2-yl, optionally substituted pyrazol-4-yl, optionally substituted pyridin-3-yl, optionally substituted pyrazol-4-yl, optionally substituted 7-aza-5,6,7,8-tetrahydroindolizin-1-yl, optionally substituted pyridazin-3-yl or optionally substituted pyridin-4-yl. A is an optionally substituted C2-C9 heteroaryl or a C6-C substituted by an optionally substituted C2-C9 heteroaryl. 10 In some embodiments, R A is cyclopropyl, methyl, methoxy or [ka] In some embodiments, R A is phenyl substituted with pyrazol-1-yl. In some embodiments, R A teeth, [ka] It is.

[0039] In some embodiments, R 1 is pyridin-3-yl, pyridin-4-yl or [ka] It is.

[0040] In some embodiments, the compound has the structure [ka] or a pharma- ceutically acceptable salt thereof.

[0041] In some embodiments, the compound has the structure [ka] or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound has the structure: [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] or a pharma- ceutically acceptable salt thereof.

[0042] In some embodiments, the compound has the structure: [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] or a pharma- ceutically acceptable salt thereof.

[0043] In one aspect, the invention features a pharmaceutical composition including any of the compounds described above and a pharma- ceutically acceptable excipient.

[0044] In an aspect, the invention features a method of treating a neurological disorder (e.g., frontotemporal dementia-TDP (FTLD-TDP), chronic traumatic encephalopathy, ALS, Alzheimer's disease, limbic-predominant age-related TDP-43 encephalopathy (LATE), or frontotemporal lobar degeneration) in a subject in need thereof, the method comprising administering an effective amount of any of the compounds or pharmaceutical compositions described above.

[0045] In an aspect, the invention features a method of inhibiting toxicity in a cell (e.g., a mammalian neuronal cell) associated with a protein (e.g., TDP-43 or C9orf72), the method including administering an effective amount of any of the compounds or pharmaceutical compositions described above.

[0046] In an aspect, the invention features a method of treating a TDP-43- or C9orf72-associated disorder (e.g., FTLD-TDP, chronic traumatic encephalopathy, ALS, Alzheimer's disease, LATE, or frontotemporal lobar degeneration) in a subject in need thereof. The method includes administering to the subject an effective amount of a compound described herein or a pharmaceutical composition containing one or more compounds described herein. In some embodiments, the method includes administering to the subject in need thereof an effective amount of a compound of formula 14 [ka] or a pharma- ceutically acceptable salt thereof (In the formula, [ka] is a single bond, and X 1 is (C(R A )2) m or -OC(R A )2-R X and X 2 is C(R A )2 or CO, or [ka] is a double bond and X 1 and X 2 are each independently A or N, where R X X 2 is a bond to R 1 , -(L) n -R B , hydrogen, halogen, cyano, optionally substituted C 1~6 Alkyl, optionally substituted C 1~6 Heteroalkyl, optionally substituted C 1~6 Alkoxy, optionally substituted C 6~10 Aryl, optionally substituted C 1~9Heterocyclyl or optionally substituted C 1~9 is heteroaryl, R 2 is hydrogen, optionally substituted C 1~6 Alkyl, optionally substituted C 6~10 Aryl, optionally substituted C 1~9 Heterocyclyl, optionally substituted C 1~9 Heteroaryl or optionally substituted C6-C 10 aryl C1-C6 alkyl; R 3 is a group having the following structure: [ka] and R A each independently represents H, optionally substituted C 1~6 Alkyl, optionally substituted C 6~10 Aryl or two geminal R A the groups, together with the atoms to which they are attached, form oxo, R B is replaced as necessary 6~10 Aryl, optionally substituted C 1~9 Heteroaryl, optionally substituted C 3~8 Cycloalkyl, -N=CH-R D or C, substituted as appropriate 1~9 heterocyclyl, optionally substituted C2-C9 heteroarylC1-C6 alkyl, optionally substituted C2-C9 heterocyclylC1-C6 alkyl; R C is H or optionally substituted C1-C6 alkyl; R D is optionally substituted C6-C 10 is aryl, Each L is independently an optionally substituted alkylene, an optionally substituted C1-C6 heteroalkylene, an optionally substituted C3-C8 cycloalkylene, an optionally substituted C2-C6 alkynylene, O, or NR C and n is 1, 2 or 3; m is 0, 1 or 2. The method includes administering

[0047] In some embodiments, [ka] is a single bond. In some embodiments, X 1 is (C(R A )2) m In some embodiments, m is 1. In some embodiments, X 2 is C(R A In some embodiments, R A are each hydrogen.

[0048] In an aspect, the invention features a method of inhibiting PIKfyve, the method including contacting a cell with an effective amount of any of the compounds or pharmaceutical compositions described above.

[0049] In another aspect, the present invention features a method of treating a neurological disorder in a patient, such as a human patient, who is identified as likely to benefit from treatment with a compound of the present invention based on TDP-43 toxicity.In this aspect, the method can include (i) determining that the patient exhibits or is prone to develop TDP-43 toxicity, and (ii) providing the patient with a therapeutically effective amount of a compound of the present invention.In some embodiments, the patient has previously been determined to exhibit or be prone to develop TDP-43 toxicity, and the method includes providing the patient with a therapeutically effective amount of a compound of the present invention.The susceptibility of a patient to developing TDP-43 aggregation can be determined, for example, by determining whether the patient expresses a mutant isoform of TDP-43, which includes a mutation associated with TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D. This can be done, for example, by determining the amino acid sequence of a TDP-43 isoform isolated from a sample obtained from the patient, or by determining the nucleic acid sequence of a TDP-43 gene isolated from a sample obtained from the patient. In some embodiments, the method includes obtaining a sample from the patient.

[0050] In a further aspect, the present invention features a method for treating a neurological disorder in a patient, such as a human patient, identified based on TDP-43 expression as likely to benefit from treatment with a compound of the present invention.In this aspect, the method includes (i) determining that the patient expresses a mutant form of TDP-43 having a mutation associated with TDP-43 aggregation (e.g., a mutation selected from Q331K, M337V, Q343R, N345K, R361S and N390D), and (ii) providing the patient with a therapeutically effective amount of a compound of the present invention.In some embodiments, the patient has previously been determined to express a mutant form of TDP-43 having a mutation associated with TDP-43 aggregation, such as a Q331K, M337V, Q343R, N345K, R361S or N390D mutation, and the method includes providing the patient with a therapeutically effective amount of a compound of the present invention.

[0051] In another aspect, the invention features a method for determining whether a patient (e.g., a human patient) with a neurological disorder is likely to benefit from treatment with a compound of the invention by (i) determining whether the patient exhibits or is prone to develop TDP-43 aggregation, and (ii) identifying the patient as likely to benefit from treatment with a compound of the invention if the patient exhibits or is prone to develop TDP-43 aggregation. In some embodiments, the method further includes (iii) informing the patient whether the patient is likely to benefit from treatment with a compound of the invention. The susceptibility of a patient to developing TDP-43 aggregation can be determined, for example, by determining whether the patient expresses a mutant isoform of TDP-43, including a mutation associated with TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D. This can be done, for example, by determining the amino acid sequence of a TDP-43 isoform isolated from a sample obtained from the patient, or by determining the nucleic acid sequence of a TDP-43 gene isolated from a sample obtained from the patient. In some embodiments, the method includes obtaining a sample from the patient.

[0052] In another aspect, the invention features a method for determining whether a patient (e.g., a human patient) with a neurological disorder is likely to benefit from treatment with a compound of the invention by (i) determining whether the patient expresses a TDP-43 mutant having a mutation associated with TDP-43 aggregation (e.g., a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D), and (ii) identifying the patient as likely to benefit from treatment with a compound of the invention if the patient expresses a TDP-43 mutant. In some embodiments, the method further includes (iii) informing the patient whether the patient is likely to benefit from treatment with a compound of the invention. The TDP-43 isoform expressed by the patient can be assessed, for example, by isolating TDP-43 protein from a sample obtained from the patient and sequencing the protein using molecular biology techniques described herein or known in the art. In some embodiments, the TDP-43 isoform expressed by the patient is determined by analyzing the patient's genotype at the TDP-43 locus, for example, by determining the sequence of the TDP-43 gene in a sample obtained from the patient. In some embodiments, the method includes obtaining a sample from the patient.

[0053] In some embodiments of any of the above aspects, the compound of the invention is provided to a patient by administering a compound of the invention to the patient. In some embodiments, the compound of the invention is provided to a patient by administration of a prodrug that is converted in vivo to the compound of the invention.

[0054] In some embodiments of any of the above aspects, the neurological disorder is a neuromuscular disorder, such as a neuromuscular disorder selected from amyotrophic lateral sclerosis, congenital myasthenic syndromes, congenital myopathies, cramp fasciculation syndrome, Duchenne muscular dystrophy, glycogen storage disease type II, hereditary spastic paraplegia, inclusion body myositis, Isaacs syndrome, Kearns-Sayre syndrome, Lambert-Eaton myasthenic syndrome, mitochondrial myopathy, muscular dystrophy, myasthenia gravis, myotonic dystrophy, peripheral neuropathy, spinal-bulbar muscular atrophy, spinal muscular atrophy, stiff-person syndrome, Troyer syndrome, and Guillain-Barre syndrome. In some embodiments, the neurological disorder is amyotrophic lateral sclerosis.

[0055] In some embodiments of any of the above aspects, the neurological disorder is selected from frontotemporal degeneration (also referred to as frontotemporal lobar degeneration and frontotemporal dementia), Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, corticobasal degeneration, progressive supranuclear palsy, Guam dementia-parkinsonism ALS complex, Huntington's disease, early-onset Paget's disease and inclusion body myopathy with frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander's disease, and hereditary inclusion body myopathy.

[0056] In some embodiments, the neuropathy is amyotrophic lateral sclerosis, and following administration of a compound of the invention to the patient, the patient exhibits one or more, or all of the following responses: (i) an improvement in a condition as assessed using the ALSFRS or ALSFRS-R, such as an improvement in a patient's Amyotrophic Lateral Sclerosis Functional Rating Scale (ALSFRS) or Revised ALSFRS (ALSFRS-R) score within one day or more days, weeks, or months after administration of a compound of the invention (e.g., within about one day to about 48 weeks (e.g., within about two days to about 36 weeks, about four weeks to about 24 weeks, about eight weeks to about 20 weeks, or about twelve weeks to about 16 weeks) or more weeks after the patient's first administration of a compound of the invention, e.g., within one day, two days, three days, four days, five days, or more days after the patient's first administration of a compound of the invention) improvement in the patient's ALSFRS or ALSFRS-R score within 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, or more (ii) an improvement in slow vital capacity, such as an improvement in a patient's slow vital capacity within one or more days, weeks, or months after administration of a compound of the invention (e.g., within about one day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the patient is first administered a compound of the invention, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 1 week ... Improvement in the patient's normal lung capacity within 1 week, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, or more (iii) a reduction in the reduced response exhibited by the patient upon repeated nerve stimulation, such as a reduction observed within one day or more days, weeks, or months after administration of a compound of the invention (e.g., within about one day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after initial administration of a compound of the invention to the patient, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks after initial administration of a compound of the invention to the patient) , a decrease observed within 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 weeks or more), (iv) an improvement in muscle strength as assessed, for example, by the UK Medical Research Council Muscle Testing Scale (related to measuring patient response to treatment of a nervous system disease, the disclosure of which is incorporated herein by reference), such as an improvement observed within one day or more days, weeks, or months after administration of a compound of the invention (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the initial administration of a compound of the invention to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, or more weeks after the initial administration of a compound of the invention to the patient). , improvement observed within 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks or more), (v) an improvement in quality of life, such as an improvement in the patient's quality of life, as assessed, for example, using an amyotrophic lateral sclerosis specific quality of life (ALS specific QOL) questionnaire, observed within one day or more days, weeks, or months after administration of a compound of the invention (e.g., within about one day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the patient's first administration of a compound of the invention, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 days, 45 days, 46 days, 47 days, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 an improvement in the subject's quality of life observed within days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, or more; (vi) a reduction in the frequency and / or severity of muscle cramps, such as a reduction in cramp frequency and / or severity within 1 day or more days, weeks, or months after administration of a compound of the invention (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the initial administration of a compound of the invention to the patient, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks after the initial administration of a compound of the invention to the patient , 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 weeks or more); and / or a reduction in seizure frequency and / or severity within (vii) a reduction in TDP-43 aggregation, such as a reduction in TDP-43 aggregation within 1 day or more days, weeks, or months after administration of a compound of the invention (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the initial administration of a compound of the invention to a patient, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks after the initial administration of a compound of the invention to a patient , 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks or more reduction in TDP-43 aggregation). chemical terms

[0057] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0058] Those of skill in the art will recognize that certain compounds described herein may exist in one or more different isomeric (e.g., stereoisomers, geometric isomers, tautomers), and / or isotopic (e.g., one or more atoms replaced by a different isotope of that atom, such as hydrogen being replaced by deuterium) forms. Unless otherwise indicated or apparent from the context, it will be understood that the depicted structures represent all such isomeric or isotopic forms individually or in combination.

[0059] In some embodiments, one or more compounds depicted herein may exist in different tautomeric forms.Unless otherwise expressly excluded, when such compounds are mentioned, all such tautomeric forms are included.In some embodiments, tautomeric forms result from the exchange of a single bond with an adjacent double bond and the concomitant migration of a proton.In certain embodiments, tautomeric forms may be prototropic tautomers, which are isomeric protonation states that have the same empirical formula and total charge as the reference form. Examples of moieties that contain prototropic tautomeric forms are ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and ring forms in which protons can occupy two or more positions in the heterocyclic ring system, such as 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole.In some embodiments, the tautomeric forms can be in equilibrium or can be sterically fixed in one form by appropriate substitution.In certain embodiments, the tautomeric forms result from acetal interconversions, such as the interconversions illustrated in the following scheme: [ka]

[0060] Those skilled in the art will recognize that in some embodiments, isotopes of the compounds described herein can be prepared and / or utilized by the present invention. "Isotopes" refers to atoms that have the same atomic number but have different mass numbers due to different numbers of neutrons in the nucleus. For example, isotopes of hydrogen include tritium and deuterium. In some embodiments, isotopic substitution (e.g., substitution of hydrogen with deuterium) can modify the physicochemical properties of a molecule, such as the rate of metabolism and / or racemization of chiral centers.

[0061] As is known in the art, many chemical entities (particularly many organic molecules and / or many small molecules) can be in a variety of different solid forms, such as, for example, amorphous and / or crystalline forms (e.g., polymorphs, hydrates, solvates, etc.). In some embodiments, such entities may be utilized in any form, including any solid form. In some embodiments, such entities are utilized in a particular form, e.g., a particular solid form.

[0062] In some embodiments, the compounds described and / or illustrated herein may be provided and / or utilized in a salt form.

[0063] In certain embodiments, the compounds described and / or illustrated herein may be provided and / or utilized in a hydrated or solvated form.

[0064] Substituents of the compounds of the present disclosure are disclosed herein in groups or ranges at various positions. It is specifically intended that the present disclosure includes all individual subcombinations of the members of such groups and ranges. For example, the term "C1-C6 alkyl" is specifically intended to individually disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl. Furthermore, when a compound includes multiple positions where a substituent is disclosed in a group or range, unless otherwise indicated, the present disclosure is intended to encompass the individual compounds and groups (e.g., classes and subclasses) of compounds, including all individual subcombinations of the members at each position.

[0065] As used herein, phrases of the form "optionally substituted X" (e.g., optionally substituted alkyl) are intended to be equivalent to "X, where X is optionally substituted, X" (e.g., "alkyl, where the alkyl is optionally substituted, alkyl"). The feature "X" (e.g., alkyl) is not itself intended to imply optionality.

[0066] The term "acyl," as used herein, refers to a hydrogen, alkyl, aryl, or heteroaryl, as defined herein, attached to the parent molecular group through a carbonyl group, and includes, by way of example, formyl (i.e., a carboxaldehyde group), acetyl, trifluoroacetyl, propionyl, butanoyl, and benzoyl. Exemplary unsubstituted acyl groups contain 1 to 6, 1 to 11, or 1 to 21 carbons. Optionally substituted acyl includes:

[0067] The term "alkyl," as used herein, refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon radical of 1 to 20 carbon atoms (e.g., 1 to 16 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms). Alkylene is a divalent alkyl group.

[0068] The term "alkenyl," as used herein, alone or in combination with other groups, refers to a straight-chain or branched hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms).

[0069] The term "alkynyl," as used herein, alone or in combination with other groups, refers to a straight-chain or branched hydrocarbon residue having a carbon-carbon triple bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms). Alkynylene is a divalent alkynyl group.

[0070] The term "amino" as used herein refers to -N(R N1 )2, R N1 are independently H, OH, NO2, and N(R N2 )2, SO2OR N2 , SO2R N2 , SOR N2, an N-protecting group, alkyl, alkoxy, aryl, arylalkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), and these described R N1 Each group may be optionally substituted or may have two R N1 taken together form an alkylene or heteroalkylene, R N2 is each independently H, alkyl, or aryl. The amino group of the present invention can be an unsubstituted amino (i.e., -NH) or a substituted amino (i.e., -N(R N1 )2).

[0071] The term "aryl" as used herein refers to an aromatic carbomonocyclic or polycyclic radical of 6 to 12 carbon atoms having at least one aromatic ring. Examples of such groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indanyl, and 1H-indenyl.

[0072] The term "arylalkyl," as used herein, refers to an alkyl group substituted with an aryl group. Exemplary unsubstituted arylalkyl groups are those having 7 to 30 carbons (e.g., C 1, C 2, C 3, C 4, C 5, C 6, C 7, C 8, C 9, C 10, C 11, C 12, C 13, C 14, C 15, C 16, C 17, C 18, C 19, C 20, C 21, C 22, C 23, C 24, C 25, C 26, C 27, C 28, C 29, C 30, C 31, C 28, C 32, C 29, C 33, C 34, C 35, C 36, C 37, C 38, C 39, C 40, C 41 6~10 Aryl C1-C6 alkyl, C 6~10 Aryl C1-C 10 Alkyl or C 6~10 Aryl C1-C 20 In some embodiments, the alkyl and aryl may each be further substituted with one, two, three, or four substituents, as defined herein for each group.

[0073] The term "azido" as used herein refers to the group --N3.

[0074] The term "cyano" as used herein refers to the group CN.

[0075] The term "carbocyclyl" as used herein refers to a non-aromatic C-C ring in which the ring is formed by carbon atoms. 12 It refers to a monocyclic, bicyclic or tricyclic structure. The carbocyclyl structure comprises a cycloalkyl group and an unsaturated carbocyclyl radical.

[0076] The term "cycloalkyl" as used herein refers to a saturated, non-aromatic, monovalent, carbocyclic or polycyclic radical having from 3 to 10, preferably from 3 to 6, carbon atoms. This term is further exemplified by radicals such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl. Cycloalkylene is a divalent cycloalkyl group.

[0077] The term "halo," as used herein, means a fluorine (fluoro), chlorine (chloro), bromine (bromo) or iodine (iodo) radical.

[0078] The term "heteroalkyl" as used herein refers to an alkyl group, as defined herein, in which one or more of the constituent carbon atoms are replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkyl group can be further substituted with one, two, three, or four substituents as described herein for alkyl groups. An example of a heteroalkyl group is "alkoxy," which as used herein refers to alkyl-O- (e.g., methoxy and ethoxy). Heteroalkylene is a divalent heteroalkyl group.

[0079] The term "heteroalkenyl" as used herein refers to an alkenyl group as defined herein, in which one or more constituent carbon atoms are replaced by nitrogen, oxygen or sulfur. In some embodiments, the heteroalkenyl group can be further substituted with one, two, three or four substituents as described herein for alkenyl groups. An example of a heteroalkenyl group is "alkenoxy", which refers to alkenyl-O- as used herein. Heteroalkenylene is a divalent heteroalkenyl group.

[0080] The term "heteroalkynyl" as used herein refers to an alkynyl group as defined herein, in which one or more of the constituent carbon atoms are replaced by nitrogen, oxygen or sulfur. In some embodiments, the heteroalkynyl group can be further substituted with one, two, three or four substituents as described herein for alkynyl groups. An example of a heteroalkynyl group is "alkynoxy", which as used herein refers to alkynyl-O-. Heteroalkynylene is a divalent heteroalkynyl group.

[0081] The term "heteroaryl" as used herein refers to an aromatic monocyclic or polycyclic radical of 5 to 12 atoms having at least one aromatic ring and containing 1, 2, 3 or 4 ring heteroatoms selected from N, O and S, the remaining ring atoms being C. One or two ring carbon atoms of the heteroaryl group may be replaced by a carbonyl group. Examples of heteroaryl groups are pyridyl, pyrazoyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, oxaxolyl and thiazolyl.

[0082] The term "heteroarylalkyl," as used herein, refers to an alkyl group substituted with a heteroaryl group. Exemplary unsubstituted heteroarylalkyl groups include those having 7 to 30 carbons (e.g., C2-C9 heteroaryl C1-C6 alkyl, C2-C9 heteroaryl C1-C 10 Alkyl or C2-C9 heteroaryl C1-C 20 In some embodiments, alkyl and heteroaryl may each be further substituted with 1, 2, 3, or 4 substituents, as defined herein for each group.

[0083] The term "heterocyclyl," as used herein, refers to a monocyclic or polycyclic radical having 3 to 12 atoms, with at least one ring containing 1, 2, 3, or 4 ring heteroatoms selected from N, O, or S, and none of the rings being aromatic. Examples of heterocyclyl groups include, but are not limited to, morpholinyl, thiomorpholinyl, furyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl, and 1,3-dioxanyl.

[0084] The term "heterocyclylalkyl," as used herein, refers to an alkyl group substituted with a heterocyclyl group. Exemplary unsubstituted heterocyclylalkyl groups include those having 7 to 30 carbons (e.g., C2-C9 heterocyclylC1-C6 alkyl, C2-C9 heterocyclylC1-C 10 Alkyl or C2-C9 heterocyclyl C1-C 20 In some embodiments, the alkyl and heterocyclyl each may be further substituted with 1, 2, 3, or 4 substituents, as defined herein for each group.

[0085] The term "hydroxyl" as used herein refers to an --OH group.

[0086] The term "N-protecting group" as used herein refers to a group intended to protect an amino group from undesired reactions during synthetic procedures. Commonly used N-protecting groups are described in Greene, "Protective Groups in Organic Synthesis," 3 rdEdition (John Wiley & Sons, New York, 1999). N-protecting groups include acyl, aryloyl or carbamyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and chiral auxiliaries such as protected or unprotected D-amino acids, L-amino acids or D,L-amino acids such as alanine, leucine and phenylalanine; sulfonyl-containing groups such as benzenesulfonyl and p-toluenesulfonyl; benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl, 2,5-dimethoxybenzyloxycarbonyl, 2,6-dimethoxybenzyloxycarbonyl, 2,7-dimethoxybenzyloxycarbonyl, 2,8-dimethoxybenzyloxycarbonyl, 2,9-dimethoxybenzyloxycarbonyl, 3,2-dimethoxybenzyloxycarbonyl, 3,2-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,5-dimethoxybenzyloxycarbonyl, 2,6-dimethoxybenzyloxycarbonyl, 2,7-dimethoxybenzyloxycarbonyl, 2,8-dimethoxybenzyloxycarbonyl, 2,9-dimethoxybenzyloxycarbonyl, 2,9-dimethoxybenzyloxycarbonyl, 2,10-dimethoxybenzyloxycarbonyl, 2,10-dimethoxybenzyloxycarbonyl, 2,1 oxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl carbamate-forming groups such as aryl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, and phenylthiocarbonyl; arylalkyl groups such as benzyl, triphenylmethyl, and benzyloxymethyl, and silyl groups such as trimethylsilyl.Preferred N-protecting groups are alloc, formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, t-butyloxycarbonyl (Boc) and benzyloxycarbonyl (Cbz).

[0087] The term "nitro" as used herein refers to a NO2 group.

[0088] The term "oxyheteroaryl," as used herein, refers to a heteroaryl group having at least one ring oxygen atom.

[0089] The term "oxyheterocyclyl," as used herein, refers to a heterocyclyl group having at least one ring oxygen atom.

[0090] The term "thiol" as used herein refers to a --SH group.

[0091] The alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl (e.g., cycloalkyl), aryl, heteroaryl and heterocyclyl groups may be substituted or unsubstituted. When substituted, 1 to 4 substituents are generally present, unless otherwise specified. Substituents include, for example: aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halo (e.g., fluoro), hydroxyl, oxo, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy or thioalkoxy), heteroaryl, heterocyclyl, amino (e.g., NH2 or mono- or dialkylamino), azido, cyano, nitro or thiol. The aryl, carbocyclyl (e.g., cycloalkyl), heteroaryl and heterocyclyl groups may also be substituted by alkyl, such as unsubstituted and substituted arylalkyl (e.g., substituted and unsubstituted benzyl).

[0092] The compounds of the present invention may have one or more asymmetric carbon atoms and may exist in the form of optically pure enantiomers, mixtures of enantiomers, such as racemates, optically pure diastereoisomers, mixtures of diastereoisomers, racemates of diastereoisomers or racemic mixtures of diastereoisomers. Optically active forms can be obtained, for example, by resolution of racemates, by asymmetric synthesis or asymmetric chromatography (chromatography using chiral adsorbents or eluents). That is, certain disclosed compounds may exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are pairs of stereoisomers, most commonly pairs of stereoisomers whose mirror images are not superimposable because they contain asymmetrically substituted carbon atoms that act as chiral centers. "Enantiomer" means one of a pair of molecules that are mirror images of each other and are not superimposable. Diastereomers are stereoisomers that are not mirror images, most commonly because they contain two or more asymmetrically substituted carbon atoms and represent the configuration of substituents around one or more asymmetric carbon atoms. Enantiomers of a compound can be prepared, for example, by separating one enantiomer from a racemate using one or more well-known techniques and methods, such as, for example, chiral chromatography and separation methods based thereon. Suitable techniques and / or methods for separating one enantiomer of a compound described herein from a racemic mixture can be easily determined by those skilled in the art. "Racemate" or "racemic mixture" refers to a compound that contains two enantiomers, and such mixtures do not exhibit optical activity, i.e., they do not rotate the plane of polarized light. "Geometric isomer" refers to isomers that differ in the orientation of substituent atoms with respect to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic system. The atoms (other than H) on each side of the carbon-carbon double bond may be in the E (substituents are on opposite sides of the carbon-carbon double bond) or Z (substituents point to the same side) configuration. * ", "R *", "E", "Z", "cis" and "trans" indicate configurations relative to the core molecule. Certain disclosed compounds may exist in the form of atropisomers. Atropisomers are stereoisomers resulting from hindered rotation about a single bond, where the steric distortion barrier to rotation is high enough to allow isolation of the conformers. The compounds of the present invention can be prepared as individual isomers either by isomer-specific synthesis or by resolution from an isomeric mixture. Conventional resolution techniques include forming a salt of the free base of each isomer of the isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming a salt of the acid form of each isomer of the isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each isomer of the isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving the isomeric mixture of either the starting material or the final product using a variety of well-known chromatographic methods. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by weight relative to the other stereoisomer. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% optically pure by weight. When a single diastereomer is named or depicted by structure, the named or depicted diastereomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% pure by weight. Percent optical purity is the ratio of the weight of an enantiomer, or the weight of an enantiomer to the sum of the weights of its enantiomer. Diastereomeric purity by weight is the ratio of the weight of one diastereomer, or the weight of all diastereomers. Where the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% pure on a mole fraction basis relative to other stereoisomers.When a single enantiomer is depicted by name or structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% pure on a mole fraction basis. When a single diastereomer is depicted by name or structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% pure on a mole fraction basis. Percent purity on a mole fraction basis is the ratio of moles of an enantiomer, or moles of an enantiomer to the sum of moles of its optical isomer. Similarly, percent purity on a mole fraction basis is the ratio of moles of a diastereomer, or moles of a diastereomer to the sum of moles of its optical isomer. When a disclosed compound is named or illustrated by structure without indicating stereochemistry and the compound has at least one chiral center, the name or structure should be understood to encompass either an enantiomer of the compound without the corresponding optical isomer, a racemic mixture of the compound, or a mixture enriched in one enantiomer relative to its corresponding optical isomer. When a disclosed compound is named or illustrated by structure without indicating stereochemistry and has two or more chiral centers, the name or structure should be understood to encompass a diastereomer without the other diastereomer, some diastereomers without other diastereomeric pairs, a mixture of diastereomers, a mixture of diastereomeric pairs, a mixture of diastereomers enriched in one diastereomer relative to the other diastereomer(s), or a mixture of diastereomers enriched in one or more diastereomers relative to the other diastereomers. The present invention encompasses all of these forms. definition

[0093] In this application, unless otherwise clear from the context, (i) the term "a" may be understood to mean "at least one," (ii) the term "or" may be understood to mean "and / or," (iii) the terms "comprising" and "including" may be understood to encompass the listed components or steps, whether presented alone or together with one or more additional components or steps, (iv) the terms "about" and "approximately" may be understood to allow for standard deviation, as understood by one of ordinary skill in the art, and (v) when ranges are presented, the endpoints are included.

[0094] As used herein, the term "administration" refers to the administration of a composition (e.g., a compound, a conjugate, or a preparation comprising a compound or conjugate described herein) to a subject or system. Administration to an animal subject (e.g., a human) can be by any suitable route. For example, in some embodiments, administration can be bronchial (including by bronchial instillation), oral, intraintestinal, intradermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, intramucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, intratracheal (including by intratracheal instillation), transdermal, intravaginal, and intravitreal.

[0095] As used herein, the term "animal" refers to any member of the animal kingdom. In some embodiments, "animal" refers to humans at any stage of development. In some embodiments, "animal" refers to non-human animals at any stage of development. In some embodiments, the non-human animals are mammals (e.g., rodents, mice, rats, rabbits, monkeys, dogs, cats, sheep, cows, primates, and / or pigs). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, and / or worms. In some embodiments, animals may be transgenic animals, genetically engineered animals, and / or clones.

[0096] As used herein, the terms "approximately" and "about" are intended to encompass normal statistical variations that would be understood by those skilled in the art, respectively, when appropriate to the relevant context.In certain embodiments, the terms "approximately" or "about" refer to a range of values ​​that falls within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (greater or less) of the stated value, respectively, unless otherwise specified or otherwise clear from the context (e.g., when such number exceeds 100% of possible values).

[0097] As used herein, the term associated refers to two events or entities being "associated" with each other if the presence, level and / or form of one correlates with that of the other. For example, a specific entity (e.g., a polypeptide) is considered to be associated with a particular disease, disorder or condition if its presence, level and / or form correlates with the incidence and / or susceptibility of the disease, disorder or condition (e.g., across a relevant population).

[0098] As used herein, the terms "benefit" and "response" are used interchangeably in the context of a subject, such as a human subject, undergoing therapy to treat neurological disorders, such as amyotrophic lateral sclerosis, frontotemporal degeneration (also referred to as frontotemporal lobar degeneration and frontotemporal dementia), Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, corticobasal degeneration, progressive supranuclear palsy, Guam dementia-parkinsonism ALS complex, Huntington's disease, early-onset Paget's disease and inclusion body myopathy with frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander's disease and hereditary inclusion body myopathy.The terms "benefit" and "response" refer to any clinical improvement in the condition of a subject. Exemplary benefits in the context of a subject undergoing treatment for a neurological disorder using the compositions and methods described herein (e.g., in the context of a human subject undergoing treatment for a neurological disorder described herein, such as amyotrophic lateral sclerosis, with a FYVE-type zinc finger-containing phosphoinositide kinase (PIKfyve) inhibitor described herein, such as an inhibitory small molecule, antibody, antigen-binding fragment thereof, or interfering RNA molecule) include slowing and halting disease progression, and suppression of one or more symptoms associated with the disease. In particular, examples of clinical "benefit" and "response" in the context of a patient (e.g., a human patient) undergoing treatment for amyotrophic lateral sclerosis with a compound of the invention include (i) an improvement in the subject's Amyotrophic Lateral Sclerosis Functional Rating Scale (ALSFRS) or Revised ALSFRS (ALSFRS-R) score within one or more days, weeks, or months following administration of a compound of the invention, as assessed using the ALSFRS or ALSFRS-R after administration of a compound of the invention. Improvement in the subject's condition (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after initial administration of a compound of the present invention to the subject, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks,(ii) an improvement in a subject's ALSFRS or ALSFRS-R score within 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks or more following administration of a compound of the invention; An improvement in the normal lung capacity of a subject following administration of a compound of the invention, such as an improvement in the normal lung capacity of a subject within a number of days, weeks, or months (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the initial administration of a compound of the invention to the subject, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks or more); (iii) an improvement in the normal lung capacity of a subject within one or more days following administration of a compound of the invention upon repetitive nerve stimulation; A reduction in the attenuation response exhibited by the subject, such as a reduction observed within a greater number of days, weeks, or months (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the initial administration of a compound of the present invention to the subject, such as within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks,(iv) a reduction observed within 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks or more following administration of a compound of the invention), e.g., an improvement observed within one or more days, weeks or months following administration of a compound of the invention, e.g., as measured by the British Medical Research Council Muscle Test Scale (related to the measurement of patient response to treatment of nervous system disorders, the disclosure of which is incorporated herein by reference, e.g., Jagtap et al., Ann. Indian. Acad. Neurol. 17:336-339 (2014)) or more weeks after the initial administration of a compound of the present invention to the subject, e.g., within 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the initial administration of a compound of the present invention to the subject, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, (v) an improvement in the subject's quality of life, such as an improvement observed within 1 day or more days, weeks, or months following administration of a compound of the invention, as assessed, for example, using an amyotrophic lateral sclerosis specific quality of life (ALS specific QOL) questionnaire (e.g., an improvement observed within about 1 day to about 48 weeks (e.g., ...Within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks, for example, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 8 (vi) an improvement in the quality of life of the subject observed within 1 week, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks or more following administration of a compound of the invention; and (vi) an improvement in the frequency and / or severity of seizures exhibited by the subject within 1 day or more days, weeks or months following administration of a compound of the invention. a reduction in the frequency and / or severity of muscle spasms, such as a reduction in the severity of the spasms (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the initial administration of a compound of the invention to the subject, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 weeks or more).

[0099] As used herein, the term "dosage form" refers to a physically discrete unit of an active compound (e.g., a therapeutic or diagnostic agent) for administration to a subject. Each unit contains a predetermined amount of active agent. In some embodiments, such amount is a unit dosage (or a fraction thereof) suitable for administration according to a dosing regimen (i.e., a therapeutic dosing regimen) determined to correlate with a desired or beneficial outcome when administered to a relevant population. Those skilled in the art will recognize that the total amount of a therapeutic composition or compound to be administered to a particular subject is determined by one or more attending physicians and may include administration of multiple dosage forms.

[0100] As used herein, the term "dosing regimen" refers to a set of unit doses (usually more than one) that are administered individually to a subject, usually separated by a period of time. In some embodiments, a given therapeutic compound has a recommended dosing regimen that can include one or more doses. In some embodiments, a dosing regimen includes multiple doses, each of which is separated from the other by a period of the same length. In some embodiments, a dosing regimen includes multiple doses and at least two different periods that separate the individual doses. In some embodiments, all doses in a dosing regimen are the same unit dose amount. In some embodiments, different doses in a dosing regimen are different amounts. In some embodiments, a dosing regimen includes a first administration at a first dosage amount, followed by one or more additional administrations at a second dosage amount that is different from the first dosage amount. In some embodiments, a dosing regimen includes a first administration at a first dosage amount, followed by one or more additional administrations at a second dosage amount that is the same as the first dosage amount. In some embodiments, the dosing regimen correlates with a desired or beneficial outcome when administered to an entire relevant population (ie, is a therapeutic dosing regimen).

[0101] In practicing the methods of this invention, an "effective amount" of any one of the compounds of this invention, or any combination of compounds of this invention or pharma- ceutically acceptable salts thereof, either alone or in combination, is administered by any of the conventional and accepted methods known in the art.

[0102] The term "pharmaceutical composition" as used herein refers to a composition that contains a compound described herein, which is formulated with pharmaceutical acceptable additives and is manufactured or sold as part of a therapeutic regimen for treating disease in a mammal with the approval of a government regulatory agency.The pharmaceutical composition can be formulated, for example, for oral administration in unit dosage form (e.g., tablet, capsule, caplet, gel cap or syrup), for topical administration (e.g., cream, gel, lotion or ointment), for intravenous administration (e.g., as a sterile solution in a solvent system suitable for intravenous use without particulate obstruction), or in any other pharmaceutical acceptable formulation.

[0103] "Pharmaceutically acceptable additives" as used herein refers to any component other than the compounds described herein (e.g., vehicle capable of suspending or dissolving active compounds), which has substantially non-toxic and non-inflammatory properties in patients.Additives can include, for example, anti-adhesive agents, antioxidants, binders, coating agents, compression aids, disintegrants, dyes (coloring agents), emollients, emulsifiers, fillers (diluents), film-forming agents or coating agents, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspending or dispersing agents, sweeteners and wetting water. Exemplary additives may include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscaromellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0104] As used herein, the term "pharmaceutical acceptable salt" refers to any pharmaceutically acceptable salt of the compound of formula (I). For example, any pharmaceutically acceptable salt of the compound described herein includes salts that are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reaction, within the scope of reasonable medical judgment, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (Eds. PH Stahl and CG Wermuth), Wiley-VCH, 2008. Salts can be prepared separately during the final isolation and purification of the compounds described herein, or in situ by reacting the free base group with a suitable organic acid.

[0105] The compounds of the present invention can have ionizable groups so that they can be prepared as pharmaceutically acceptable salts.These salts can be acid addition salts, including inorganic or organic acids, or salts can be prepared from inorganic or organic bases when the compounds of the present invention are in acidic form.In many cases, the compounds are prepared or used as pharmaceutically acceptable salts, which are prepared as addition products of pharmaceutically acceptable acids or bases.Suitable pharmaceutically acceptable acids and bases, and methods for preparing suitable salts are well known in the art.Salts can be prepared from pharmaceutically acceptable non-toxic acids and bases, including inorganic and organic acids, and inorganic and organic bases.

[0106] The terms "PIKfyve" and "FYVE-type zinc finger-containing phosphoinositide kinase" are used interchangeably herein and refer to an enzyme that catalyzes the phosphorylation of phosphatidylinositol 3-phosphate to produce phosphatidylinositol 3,5-bisphosphate, for example, in a human subject. The terms "PIKfyve" and "FYVE-type zinc finger-containing phosphoinositide kinase" refer not only to the wild-type form of PIKfyve, but also to variants of wild-type PIKfyve protein and nucleic acids encoding same. The gene encoding PIKfyve can be obtained under NCBI reference sequence number NG_021188.1. Exemplary transcript sequences of the wild-type form of human PIKfyve can be obtained under NCBI reference sequence numbers NM_015040.4, NM_152671.3 and NM_001178000.1. Exemplary protein sequences for the wild-type form of human PIKfyve are available at NCBI reference sequence numbers NP_055855.2, NP_689884.1, and NP_001171471.1.

[0107] As used herein, the term "PIKfyve inhibitor" refers to a substance such as a compound of formula I. This type of inhibitor can competitively inhibit PIKfyve activity, for example, by specifically binding to the PIKfyve enzyme (e.g., due to the affinity of the inhibitor to the PIKfyve active site), thereby disabling, hindering or stopping the entry of one or more endogenous substrates of PIKfyve into the enzyme active site. Additional examples of PIKfyve inhibitors that suppress the activity of PIKfyve enzyme include substances that can bind to PIKfyve at a site distal to the active site, and can weaken the binding of endogenous substrates to the PIKfyve active site by changing the spatial conformation of the enzyme due to the binding of the inhibitor. The term "PIKfyve inhibitor" includes substances that modulate PIKfyve activity, as well as substances that reduce the concentration and / or stability of PIKfyve mRNA transcripts in vivo, and substances that suppress the translation of functional PIKfyve enzyme.

[0108] The term "pure" means substantially pure or free of undesirable components (e.g., other compounds and / or other components of a cell lysate), contaminants, impurities or imperfections.

[0109] Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogensulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobacillus acid salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium, as well as non-toxic ammonium, quaternary ammonium, and amine cations, including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.

[0110] Various clinical indicators can be used to identify patients as "at risk" for developing certain neurological disorders: amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, corticobasal degeneration, progressive supranuclear palsy, Guam dementia-parkinsonism ALS complex, Huntington's disease, early-onset Paget's disease and inclusion body myopathy with frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander disease and hereditary inclusion body myopathy. Examples of patients (e.g., human patients) "at risk" of developing a nervous system disorder such as amyotrophic lateral sclerosis include (i) subjects exhibiting or prone to exhibiting TAR-DNA binding protein (TDP)-43 aggregation, and (ii) subjects expressing mutant forms of TDP-43 that contain mutations associated with TDP-43 aggregation and toxicity, such as mutations selected from Q331K, M337V, Q343R, N345K, R361S, and N390D. A subject "at risk" of developing amyotrophic lateral sclerosis may exhibit one or both of these characteristics, for example, prior to the first administration of a PIKfyve inhibitor according to the compositions and methods described herein.

[0111] As used herein, the terms "TAR-DNA binding protein-43" and "TDP-43" are used interchangeably and refer to a transcriptional repressor protein involved in modulating HIV-1 transcription and alternative splicing of the pre-mRNA transcript of cystic fibrosis transmembrane conductance regulator (CFTR), for example, in human subjects. The terms "TAR-DNA binding protein-43" and "TDP-43" refer not only to the wild-type form of TDP-43, but also to variants of wild-type TDP-43 protein and the nucleic acid encoding same. The amino acid sequence of the wild-type form of human TDP-43 and the corresponding mRNA sequence are provided in NCBI reference sequence numbers NM_007375.3 and NP_031401.1, respectively.

[0112] The terms "TAR-DNA binding protein-43" and "TDP-43", as used herein, include, for example, forms of the human TDP-43 protein having an amino acid sequence at least 85% identical to the amino acid sequence of NCBI Reference SEQ ID NO: NP_031401.1 (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9% or 100% identical to the amino acid sequence of NCBI Reference SEQ ID NO: NP_031401.1), and / or forms of the human TDP-43 protein that include one or more substitutions, insertions and / or deletions (e.g., one or more conservative and / or non-conservative amino acid substitutions, such as up to 5, 10, 15, 20, 25 or more conservative or non-conservative amino acid substitutions) compared to the wild-type TDP-43 protein. Patients who can be treated for neurological disorders described herein, such as, for example, amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, corticobasal degeneration, progressive supranuclear palsy, dementia-parkinsonism-ALS complex of Guam, Huntington's disease, early-onset Paget's disease and inclusion body myopathy with frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander's disease, and hereditary inclusion body myopathy, include human patients who express a form of TDP-43 having a mutation associated with increased TDP-43 aggregation and toxicity, such as a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D. Similarly, the terms "TAR-DNA binding protein-43" and "TDP-43" as used herein include forms of the human TDP-43 gene that encode an mRNA transcript having a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of, for example, NCBI Reference SEQ ID NO: NM_007375.3 (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9% or 100% identical to the amino acid sequence of, for example, NCBI Reference SEQ ID NO: NM_007375.3).

[0113] As used herein, the term "subject" refers to any organism to which the composition according to the present invention may be administered, for example, for experimental, diagnostic, preventive and / or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates and humans). A subject may be a human or animal that may seek or require treatment, may request treatment, may currently be undergoing treatment, may be undergoing treatment in the future, or may be under the supervision of a trained professional for a particular disease or condition.

[0114] "Therapeutic regimen" refers to a dosing regimen, the administration of which across a relevant population correlates with a desired or beneficial therapeutic outcome.

[0115] The term "therapeutically effective amount" refers to an amount that is sufficient to treat a disease, disorder, and / or condition when administered to a population suffering from or susceptible to the disease, disorder, and / or condition according to a therapeutic administration regimen. In some embodiments, a therapeutically effective amount is an amount that reduces the incidence and / or severity of a disease, disorder, and / or condition and / or delays the onset of one or more symptoms thereof. Those skilled in the art will appreciate that the term "therapeutically effective amount" does not in fact require that successful treatment is achieved in a particular individual. Rather, a therapeutically effective amount may be an amount that, when administered to a patient in need of such treatment, results in a particular desired pharmacological response in a significant number of subjects. It is specifically understood that a particular subject may in fact be "refractory" to a "therapeutically effective amount". By way of example, a refractory subject may have a low bioavailability such that clinical efficacy cannot be achieved. In some embodiments, reference to a therapeutically effective amount may be a reference to the amount measured in one or more particular tissues (e.g., tissues affected by a disease, disorder, or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). One of skill in the art will appreciate that in some embodiments, a therapeutically effective amount may be formulated and / or administered in a single dose. In some embodiments, a therapeutically effective amount may be formulated and / or administered in multiple doses, for example, as part of a dosing regimen. [Brief description of the drawings]

[0116] [Figure 1] Figure 1 is a scheme showing the approach to generate a control TDP-43 yeast model (FAB1 TDP-43). The control yeast TDP-43 model was generated by integrating the human TDP-43 gene and the GAL1 promoter into the yeast genome. The yeast orthologue of human PIKFYVE is FAB1.

[0117] [Diagram 2]Figure 2 is a scheme showing the approach to generate the humanized PIKFYVE TDP-43 yeast model (PIKFYVE TDP-43). The FAB1 gene was expressed by homologous recombination with a G418 resistance cassette (fab1::G418R) (Figure 2). PIKFYVE was cloned downstream of the GPD promoter harbored in a URA3-containing plasmid and introduced into the fab1::G418R ura3 strain. The pGAL1-TDP-43 construct was then introduced into the "humanized" yeast strain to assess cytotoxicity.

[0118] [Diagram 3] FIG. 3 is a histogram generated from a flow cytometry-based viability assay of FAB1 TDP-43.

[0119] [Figure 4] Figure 4 shows histograms generated from flow cytometry-based viability assays of PIKFYVE TDP-43. Upon TDP-43 induction, there was a significant increase in nonviable cells (right-most population), with a more pronounced effect for PIKFYVE TDP-43 than for the FAB1 TDP-43 line (see Figure 3).

[0120] [Diagram 5] FIG. 5 is an overlay of histograms generated from flow cytometry-based viability assays of FAB1 TDP-43 in the presence of APY0201.

[0121] [Figure 6] FIG. 6 is an overlay of histograms generated from a flow cytometry-based viability assay of PIKFYVE TDP-43 in the presence of APY0201.

[0122] [Figure 7] FIG. 7 is a scatter plot comparing the PIKFYVE TDP-43 cytoprotective efficacy and PIKfyve inhibitory activity of test compounds. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0123] Detailed Description The invention features compositions and methods for treating, inter alia, amyotrophic lateral sclerosis and other neuromuscular disorders, as well as neurological disorders such as frontotemporal degeneration, Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, corticobasal degeneration, progressive supranuclear palsy, Guam dementia-parkinsonism ALS complex, Huntington's disease, early-onset Paget's disease and inclusion body myopathy with frontotemporal dementia (IBMPFD), sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander's disease and hereditary inclusion body myopathy. In particular, the invention provides inhibitors of FYVE-type zinc finger-containing phosphoinositide kinases (PIKfyve) that can be administered to a patient (e.g., a human patient) to treat or prevent neurological disorders, such as one or more of the above-mentioned conditions. In the context of therapeutic treatment, a PIKfyve inhibitor can be administered to a patient to alleviate one or more symptoms of a disorder, such as to inhibit or prevent aggregation of TAR-DNA binding protein (TDP)-43, and / or to treat the underlying molecular pathology associated with the disease.

[0124] The disclosure herein is based in part on the discovery that PIKfyve inhibition modulates TDP-43 aggregation in cells. Suppression of TDP-43 aggregation exerts beneficial effects in patients suffering from neurological disorders. Many pathological conditions are correlated with TDP-43-promoted aggregation and toxicity, such as amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, corticobasal degeneration, progressive supranuclear palsy, Guam dementia-parkinsonism ALS complex, Huntington's disease, IBMPFD, sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander's disease and hereditary inclusion body myopathy. Without being limited by mechanism, for example, by administering an inhibitor of PIKfyve, TDP-43 aggregation induced by the PIKfyve inhibitor is suppressed, and thus patients suffering from diseases associated with TDP-43 aggregation and toxicity can be treated.

[0125] Patients who are likely to respond to PIKfyve inhibition as described herein include patients who have developed or are at risk of developing TDP-43 aggregation, such as patients who express mutant forms of TDP-43 that are associated with TDP-43 aggregation and toxicity in vivo. Examples of such mutations in TDP-43 that correlate with increased TDP-43 aggregation and toxicity include, among others, Q331K, M337V, Q343R, N345K, R361S and N390D. Thus, the compositions and methods described herein provide the additional clinical benefit of enabling identification of patients who are likely to respond to PIKfyve inhibitor therapy, and thus the process for treating these patients.

[0126] The following section provides a description of the exemplary PIKfyve inhibitors that can be used in conjunction with the compositions and methods disclosed herein.The following section further provides a description of the various exemplary administration routes and pharmaceutical compositions that can be used to deliver these substances for treating neurological disorders.

[0127] PIKfyve inhibitors Exemplary PIKfyve inhibitors described herein include compounds of formula (I): [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, [ka] is a single bond, and X 1 is (C(R A )2) m or -OC(R A )2-R X and X 2 is C(R A )2 or CO, or [ka] is a double bond and X 1 and X 2 are each independently A or N, where R X X 2 is a bond to R 1 , -(L) n -R B Halo, cyano, hydrogen, optionally substituted C 1~6 Alkoxy, optionally substituted C containing at least one ring oxygen 1~9 Heterocyclyl, optionally substituted C1-C6 alkyl, optionally substituted piperazin-1-yl, optionally substituted pyrrolidin-3-yl, pyrimidinyl (wherein the pyrimidinyl is cyclopropyl or optionally substituted C6-C 10aryl), optionally substituted pyridazinyl, optionally substituted oxazolyl, pyrid-2-on-1-yl, optionally substituted isoindolinyl, unsubstituted pyridin-4-yl, unsubstituted pyridin-2-yl, optionally substituted furan-3-yl, unsubstituted pyridin-3-yl or optionally substituted pyrazol-1-yl; R 2 is an optionally substituted C1-C6 alkyl, an optionally substituted C6-C 10 Aryl, optionally substituted piperidin-4-yl, optionally substituted tetrahydropyran-4-yl, optionally substituted pyrimidin-5-yl, optionally substituted pyrimidin-4-yl, optionally substituted pyridin-3-yl, optionally substituted pyridazin-4-yl, optionally substituted pyrazol-1-yl, optionally substituted pyrazol-4-yl, optionally substituted pyrazol-3-yl, optionally substituted pyridin-2-yl, optionally substituted triazolyl, optionally substituted benzodioxol-2-yl, optionally substituted benzodioxan-2-yl, optionally substituted C6-C 10 Aryl C1-C 10 is alkyl or optionally substituted acyl; R 3 is a group having the following structure: [ka] and R A each independently represents H, optionally substituted C 1~6 Alkyl or optionally substituted C6-C 10 Aryl or two R A together with the atom to which they are attached form an oxo, where two R A When they combine with the atom to which they are attached to form an oxo, [ka] is a single bond, R B is replaced as necessary 6~10 Aryl, optionally substituted C1-C9 heteroaryl, optionally substituted C 3~8 Cycloalkyl, -N=CH-R D or optionally substituted C1-C9 heterocyclyl, optionally substituted C2-C9 heteroaryl C1-C6 alkyl, or optionally substituted C2-C9 heterocyclyl C1-C6 alkyl; R C is H or optionally substituted C1-C6 alkyl; R D is optionally substituted C6-C 10 is aryl, or Each L is independently an optionally substituted C 1~6 Alkylene, optionally substituted C1-C6 heteroalkylene, optionally substituted C3-C8 cycloalkylene, optionally substituted C2-C6 alkynylene, O or NR C and n is 1, 2 or 3; m is 0, 1 or 2).

[0128] In some embodiments, R 1 , -(L) n -R B ;C substituted as necessary 1~6 Alkoxy; optionally substituted C containing at least one ring oxygen 1~9 heterocyclyl; unsubstituted pyrimidinyl; optionally substituted pyridazinyl; optionally substituted oxazolyl or pyrid-2-on-1-yl. 2 is replaced as necessary 6~10 Aryl, optionally substituted C 1~9Heterocyclyl or optionally substituted C 1~9 It is heteroaryl.

[0129] Exemplary PIKfyve inhibitors described herein include compounds of formula 1a: [ka] and pharma- ceutically acceptable salts thereof.

[0130] Exemplary PIKfyve inhibitors described herein include compounds of formula 1a': [ka] and pharma- ceutically acceptable salts thereof.

[0131] Exemplary PIKfyve inhibitors described herein include compounds of formula 1b: [ka] and pharma- ceutically acceptable salts thereof.

[0132] Exemplary PIKfyve inhibitors described herein include compounds of formula 1c: [ka] and pharma- ceutically acceptable salts thereof.

[0133] Exemplary PIKfyve inhibitors described herein include compounds of formula 1d: [ka] and pharma- ceutically acceptable salts thereof.

[0134] Exemplary PIKfyve inhibitors described herein include compounds of formula 1e: [ka] and pharma- ceutically acceptable salts thereof.

[0135] Exemplary PIKfyve inhibitors described herein include compounds of Formula 2: [ka] and pharma- ceutically acceptable salts thereof. (In the formula, R 2 is optionally substituted pyrimidin-3-yl or optionally substituted pyrimidin-4-yl; R 4 is hydrogen or optionally substituted C6-C 10 (aryl).

[0136] Exemplary PIKfyve inhibitors described herein include compounds of Formula 3: [ka] and pharma- ceutically acceptable salts thereof. (In the formula, R 5 is hydrogen or optionally substituted C6-C 10 is aryl, R 2 is an optionally substituted triazolyl, an optionally substituted pyrazol-4-yl, an optionally substituted pyrazol-3-yl, an optionally substituted pyrimidin-4-yl or an optionally substituted C6-C 10 aryl C1-C6 alkyl).

[0137] Exemplary PIKfyve inhibitors described herein include compounds of Formula 4: [ka] and pharma- ceutically acceptable salts thereof. (In the formula, R 2 is optionally substituted pyridin-3-yl).

[0138] Exemplary PIKfyve inhibitors described herein include compounds of Formula 5: [ka] and pharma- ceutically acceptable salts thereof. (In the formula, R 2 is optionally substituted C6-C 10 aryl; or optionally substituted pyridazin-4-yl).

[0139] Exemplary PIKfyve inhibitors described herein include compounds of Formula 6: [ka] and pharma- ceutically acceptable salts thereof. (In the formula, R 2 is optionally substituted pyridin-3-yl). Exemplary PIKfyve inhibitors described herein include compounds of Formula 7: [ka] and pharma- ceutically acceptable salts thereof. (wherein L is an optionally substituted C3-C8 cycloalkylene or C2-C6 alkynylene; R B is optionally substituted C6-C 10 (aryl).

[0140] Exemplary PIKfyve inhibitors described herein include compounds of Formula 8: [ka] and pharma- ceutically acceptable salts thereof. (In the formula, LR B is -NHN=CHR D and R 2 is optionally substituted pyridin-3-yl; R D is optionally substituted C6-C 10 (aryl).

[0141] Exemplary PIKfyve inhibitors described herein include compounds of formula 9: [ka] and pharma- ceutically acceptable salts thereof. (In the formula, R 2 is optionally substituted pyridin-3-yl; R 7 is optionally substituted C6-C 10 (aryl).

[0142] Exemplary PIKfyve inhibitors described herein include compounds of formula 10: [ka] and pharma- ceutically acceptable salts thereof. (In the formula, X 2 is N or CH, R A is an optionally substituted C2-C9 heteroaryl; or an optionally substituted C6-C 10 is aryl, R 1 is an optionally substituted C2-C9 heteroaryl or -OR 7 and R 2 is hydrogen or optionally substituted C1-C6 alkyl; R 7 is optionally substituted C2-C9 heteroaryl C1-C6 alkyl).

[0143] Additional exemplary PIKfyve inhibitors include compounds of formula 11: [ka] and pharma- ceutically acceptable salts thereof.

[0144] Exemplary PIKfyve inhibitors described herein include compounds of formula 12: [ka] and pharma- ceutically acceptable salts thereof.

[0145] Exemplary PIKfyve inhibitors described herein include compounds of formula 13: [ka] and pharma- ceutically acceptable salts thereof.

[0146] An exemplary PIKfyve inhibitor as described herein is a compound of formula 14: [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, [ka] is a single bond, and X 1 is (C(R A )2) m or -OC(R A )2-R X and X 2 is C(R A )2 or CO, or [ka] is a double bond and X 1 and X 2 are each independently A or N, where R X X 2 is a bond to R 1 , -(L) n -R B, hydrogen, halogen, cyano, optionally substituted C 1~6 Alkyl, optionally substituted C 1~6 Heteroalkyl, optionally substituted C 1~6 Alkoxy, optionally substituted C 6~10 Aryl, optionally substituted C 1~9 Heterocyclyl or optionally substituted C 1~9 is heteroaryl, R 2 is hydrogen, optionally substituted C 1~6 Alkyl, optionally substituted C 6~10 Aryl, optionally substituted C 1~9 Heterocyclyl, optionally substituted C 1~9 Heteroaryl or optionally substituted C6-C 10 aryl C1-C6 alkyl; R 3 is a group having the following structure: [ka] and R A each independently represents H, optionally substituted C 1~6 Alkyl, optionally substituted C 6~10 Aryl or two geminal R A the groups, together with the atoms to which they are attached, form oxo, R B is replaced as necessary 6~10 Aryl, optionally substituted C 1~9 Heteroaryl, optionally substituted C 3~8 Cycloalkyl, -N=CH-R D or C, substituted as appropriate 1~9 heterocyclyl, optionally substituted C2-C9 heteroarylC1-C6 alkyl, optionally substituted C2-C9 heterocyclylC1-C6 alkyl; R Cis H or optionally substituted C1-C6 alkyl; R D is optionally substituted C6-C 10 is aryl, Each L is independently an optionally substituted alkylene, an optionally substituted C1-C6 heteroalkylene, an optionally substituted C3-C8 cycloalkylene, an optionally substituted C2-C6 alkynylene, O, or NR C and n is 1, 2 or 3; m is 0, 1 or 2).

[0147] Non-limiting examples of compounds of the present invention include: [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8] [Table 3-9] [Table 3-10] [Table 3-11] [Table 3-12] [Table 3-13] and pharma- ceutically acceptable salts thereof. Treatment Inhibition of PIKfyve activity and TDP-43 aggregation to treat neurological disorders

[0148] Using the compositions and methods described herein, a patient suffering from a neurological disorder may be administered a PIKfyve inhibitor, such as a small molecule described herein, to treat the disorder and / or suppress one or more symptoms associated with the disorder. Exemplary neurological disorders that may be treated using the compositions and methods described herein include, but are not limited to, amyotrophic lateral sclerosis, frontotemporal degeneration, Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, corticobasal degeneration, progressive supranuclear palsy, dementia parkinsonism ALS complex of Guam, Huntington's disease, IBMPFD, sporadic inclusion body myositis, myofibrillar myopathy, dementia pugilistica, chronic traumatic encephalopathy, Alexander's disease and hereditary inclusion body myopathy, and congenital myasthenia. These include neuromuscular diseases such as myasthenia gravis, myotonic dystrophy, spinal and bulbar muscular atrophy, stiff-person syndrome, Troyer syndrome, and Guillain-Barré syndrome.

[0149] The present disclosure is based in part on the discovery that PIKfyve inhibitors, such as the agents described herein, can attenuate TDP-43 toxicity. TDP-43-promoted toxicity is associated with various neurological diseases. The discovery that PIKfyve inhibitors modulate TDP-43 aggregation provides important therapeutic benefits. Using PIKfyve inhibitors, such as the PIKfyve inhibitors described herein, patients suffering from neurological disorders or at risk of developing such conditions can be treated in a manner that treats the molecular pathogenesis underlying the disease. Without being limited by mechanism, the compositions and methods described herein can be used to treat or prevent such neurological conditions, for example, by inhibiting TDP-43 aggregation that promotes pathology.

[0150] Furthermore, the compositions and methods described herein provide the beneficial feature of allowing the identification and treatment of patients who are likely to respond to PIKfyve inhibitor therapy. For example, in some embodiments, a patient (e.g., a human patient suffering from or at risk of developing a nervous system disease described herein, such as amyotrophic lateral sclerosis) is administered a PIKfyve inhibitor if the patient is identified as likely to respond to such a form of treatment. Thus, a patient can be identified, for example, based on susceptibility to TDP-43 aggregation. In some embodiments, a patient is identified as likely to respond to PIKfyve inhibitor treatment based on the isoform of TDP-43 expressed by the patient. For example, a patient expressing a TDP-43 isoform with a mutation selected from Q331K, M337V, Q343R, N345K, R361S and N390D, among others, is more likely to develop TDP-43-promoted aggregation and toxicity compared to a patient who does not express such isoform of TDP-43. Using the compositions and methods described herein, patients can be identified as likely to respond to PIKfyve inhibitor therapy based on their expression of such isoforms of TDP-43, and can subsequently be administered a PIKfyve inhibitor to treat or prevent one or more neurological disorders, such as one or more neurological disorders described herein. Assessment of patient response

[0151] Various methods known in the art and described herein can be used to determine whether a patient with a neurological disorder (e.g., a patient at risk of developing TDP-43 aggregation, such as a patient expressing a mutant form of TDP-43 with a mutation associated with increased TDP-43 aggregation and toxicity, e.g., a mutation selected from Q331K, M337V, Q343R, N345K, R361S, and N390D) will respond favorably to PIKfyve inhibition. For example, successful treatment of a patient with a neurological disease, such as amyotrophic lateral sclerosis, with a PIKfyve inhibitor as described herein may be predicted by: (i) An improvement in a condition as assessed using ALSFRS or ALSFRS-R, such as an improvement in a patient's Amyotrophic Lateral Sclerosis Functional Rating Scale (ALSFRS) or Revised ALSFRS (ALSFRS-R) score within 1 day or more days, weeks, or months after administration of a PIKfyve inhibitor (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the patient's first administration of a PIKfyve inhibitor, e.g., within 1 day, 2 days, 3 days, 4 days after the patient's first administration of a PIKfyve inhibitor improvement in the patient's ALSFRS or ALSFRS-R score within 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, or more (ii) an improvement in normal lung capacity, such as an improvement in normal lung capacity of a patient within 1 day or more days, weeks, or months after administration of a PIKfyve inhibitor (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the first administration of a PIKfyve inhibitor to a patient, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, Improvement in the patient's normal lung capacity within 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 weeks or more) (iii) A decrease in the attenuation response exhibited by the patient upon repeated nerve stimulation, such as a decrease observed within 1 day or more days, weeks, or months after administration of a PIKfyve inhibitor (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the initial administration of a PIKfyve inhibitor to the patient, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks after the initial administration of a PIKfyve inhibitor to the patient). , a decrease observed within 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 weeks or more), (iv) an improvement in muscle strength as assessed, for example, by the UK Medical Research Council Muscle Test Scale (related to measuring patient response to treatment of a nervous system disease, the disclosure of which is incorporated herein by reference), such as an improvement observed within 1 day or more days, weeks, or months after administration of a PIKfyve inhibitor (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after a first administration of a PIKfyve inhibitor to a patient, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, improvement observed within 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks or more) (v) An improvement in quality of life, such as an improvement in a patient's quality of life, observed within one day or more days, weeks, or months after administration of a PIKfyve inhibitor, as assessed, for example, using an amyotrophic lateral sclerosis-specific quality of life (ALS-specific QOL) questionnaire (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the first administration of a PIKfyve inhibitor to a patient, e.g., within 1 day, 2 days, 3 days, 4 days, or more days after the first administration of a PIKfyve inhibitor to a patient). an improvement in the subject's quality of life observed within days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, or more; (vi) A reduction in the frequency and / or severity of muscle spasms, such as a reduction in spasm frequency and / or severity within 1 day or more days, weeks, or months after administration of a PIKfyve inhibitor (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the initial administration of a PIKfyve inhibitor to a patient, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks after the initial administration of a PIKfyve inhibitor to a patient). , 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 weeks or more); and / or a reduction in seizure frequency and / or severity within (vii) a reduction in TDP-43 aggregation, such as a reduction in TDP-43 aggregation within 1 day or more days, weeks, or months after administration of a PIKfyve inhibitor (e.g., within about 1 day to about 48 weeks (e.g., within about 2 days to about 36 weeks, about 4 weeks to about 24 weeks, about 8 weeks to about 20 weeks, or about 12 weeks to about 16 weeks) or more weeks after the initial administration of a PIKfyve inhibitor to a patient, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks after the initial administration of a PIKfyve inhibitor to a patient reduction in TDP-43 aggregation within 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 or more weeks). Combination preparations and their uses

[0152] The compounds of the invention can be combined with one or more therapeutic agents. In particular, the therapeutic agents can be those that treat or prophylactically treat any of the neurological disorders described herein. Combination therapy

[0153] The compounds of the present invention can be used alone or in combination with other agents that treat neuropathy or symptoms related to neuropathy, or in combination with other types of treatments that treat, prevent, and / or reduce the risk of any neuropathy.In combination treatments, the dosage of one or more therapeutic compounds may be reduced from the standard dosage when administered alone.For example, dosage may be empirically determined from drug combinations and permutations, or may be estimated by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6, 2005).In this case, the dosage of the compounds when combined should produce a therapeutic effect. Pharmaceutical Compositions

[0154] The compounds of the present invention are preferably formulated into pharmaceutical compositions for administration to human subjects in a biologically compatible form suitable for administration in vivo.Accordingly, in another aspect, the present invention provides pharmaceutical compositions comprising the compounds of the present invention in admixture with a suitable diluent, carrier or excipient.

[0155] The compounds of the present invention may be used in the form of free base, salt, solvate, and prodrug. All forms are within the scope of the present invention. According to the method of the present invention, the described compounds or salts, solvates, or their prodrugs may be administered to patients in various forms, as understood by those skilled in the art, depending on the selected route of administration. The compounds of the present invention may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, or transdermal administration, and by pharmaceutical compositions formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, pulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.

[0156] The compounds of the present invention may be administered orally, for example, with an inert diluent or an assimilable edible carrier, or the compounds of the present invention may be enclosed in a hard or soft shell gelatin capsule, or the compounds of the present invention may be compressed into tablets, or the compounds of the present invention may be incorporated directly into the food of the diet. For oral therapeutic administration, the compounds of the present invention may be incorporated into excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups and wafers.

[0157] The compounds of the present invention may also be administered parenterally. Solutions of the compounds of the present invention may be prepared in water suitably mixed with a surfactant. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof, with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2003, 20 th ed.), and The National Formulary, published in 1999 (USP 24 NF19).

[0158] The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy administration by syringe is possible.

[0159] Compositions for nasal administration can be conveniently formulated as aerosols, drops, gels and powders.Aerosol formulations usually comprise a solution or fine suspension of active substance in a physiologically acceptable aqueous or non-aqueous solvent, and are usually supplied in single or multi-dose amounts in sterile form in a sealed container, which can take the form of a cartridge or refill for use in a nebulizing device.Alternatively, the sealed container can be an integrated dispensing device, such as a single-dose nasal inhaler, or an aerosol dispenser equipped with a metering valve, which is intended to be discarded after use.When the dosage form comprises an aerosol dispenser, the dispenser comprises a propellant, which can be compressed air or a compressed gas, such as an organic propellant, such as fluorochlorohydrocarbon.Aerosol dosage forms can also take the form of a pump-action atomizer.

[0160] Compositions suitable for buccal or sublingual administration include tablets, lozenges and pastilles, in which the active ingredient is formulated with a carrier such as sugar, acacia, tragacanth, gelatin and glycerin. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base such as cocoa butter.

[0161] The compounds of the invention can be administered to animals, such as humans, either alone or in combination with pharma- ceutically acceptable carriers as described herein, the proportions of which will depend on the solubility and chemical nature of the compounds, the chosen route of administration, and standard pharmaceutical practice. Dosage

[0162] The dosage of the compounds of the present invention and / or compositions containing the compounds of the present invention may vary depending on a number of factors, such as the pharmacodynamic properties of the compound, the mode of administration, the age, health and weight of the recipient, the nature and extent of the symptoms, the frequency of treatment and the type of concurrent treatment (if any), and the clearance rate of the compound in the treated animal. Those skilled in the art can determine the appropriate dosage based on the above factors. The compounds of the present invention may be administered at a suitable dosage initially, which can be adjusted, if necessary, depending on the clinical response. In general, satisfactory results can be obtained when the compounds of the present invention are administered to humans at a daily dosage of, for example, between 0.05 mg and 3000 mg (measured as solid form). Dose ranges include, for example, between 10 and 1000 mg.

[0163] Alternatively, the dosage can be calculated using the patient's body weight. For example, the dose of the compound or pharmaceutical composition thereof administered to the patient can range from 0.1 to 50 mg / kg.

[0164] The following examples are intended to illustrate the invention. They are not intended to limit the invention in any way. EXAMPLES

[0165] [Table 4] Synthesis of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 1): [ka]

[0166] Step 1: Synthesis of 5-arylpyrimidine-2,4,6(1H,3H,5H)-trione.

[0167] To a solution of diethyl 2-allylmalonate (40.0 g, 200.0 mmol) and urea (12.0 g, 200.0 mmol) in ethanol (150 mL) was added sodium ethoxide (20% in ethanol) (80 mL) and the mixture was heated to 85 °C for 3 h. The resulting mixture was cooled to 20 °C and acetone (150 mL) was added. After stirring for 10 min, the resulting precipitate was collected by filtration, washed with petroleum ether (150 mL), and then dissolved in water (150 mL). The pH of the resulting solution was adjusted to between 3 and 4 with concentrated HCl to obtain a precipitate, which was stirred for 10 min. The solid was collected by filtration and dried under high vacuum to give 5-arylpyrimidine-2,4,6(1H,3H,5H)-trione as a brown solid (17.0 g, 51%). 1 H NMR (400 MHz, DMSO-d6) δ 11.25 (s, 2H), 5.63-5.73 (m, 1H), 5.03 (dd, J = 12.0Hz, J = 3.6Hz, 2H), 3.68 (t, J = 5.2Hz, 1H), 2.66 (t, J = 5.62Hz, 2H); LCMS (ESI) m / z: 169.1 [M+H] + .

[0168] Step 2: Synthesis of 5-allyl-2,4,6-trichloropyrimidine.

[0169] To a solution of 5-allylpyrimidine-2,4,6(1H,3H,5H)-trione (17.0 g, 101.2 mmol) in phosphorus oxychloride (60 mL) was added N,N-dimethylaniline (8.5 mL) and the solution was heated to 110° C. for 4 h. The dark solution was then cooled to 20° C. and concentrated. Ethyl acetate (300 mL) and ice water (200 mL) were added to the residue and the organic phase was separated, washed with brine (200 mL), dried and concentrated to give the crude product, which was purified by column chromatography (petroleum ether:ethyl acetate 20:1 to 10:1) to give 5-allyl-2,4,6-trichloropyrimidine as an off-white solid (16.0 g, 71%). 1H NMR (400 MHz, CDCl3) δ 5.79-5.89 (m, 1H), 5.11-5.21 (m, 2H), 3.63 (dt, J = 6.0Hz, J = 1.6Hz, 2H); LCMS (ESI) m / z: 223.1 [M+H] + .

[0170] Step 3: Synthesis of 2-(2,4,6-trichloropyrimidin-5-yl)acetaldehyde.

[0171] To a solution of 5-allyl-2,4,6-trichloropyrimidine (10.0 g, 44.7 mmol), potassium osmate(VI) dihydrate (330 mg, 0.89 mmol) and 4-methylmorpholine N-oxide (20.96 g, 89.4 mmol) in acetone (150 mL) and water (150 mL) was added sodium periodate (38.3 g, 178.8 mmol) at 0 °C and the mixture was stirred at 0-20 °C for 17 h. The resulting mixture was filtered, the filtrate was concentrated to remove acetone and the aqueous phase was extracted with ethyl acetate (150 mL × 2). The combined organic layers were washed with brine (150 mL), dried and concentrated to give the crude product. This was then purified by silica gel chromatography (petroleum ether:acetate 10:1 to 3:1) to give 2-(2,4,6-trichloropyrimidin-5-yl)acetaldehyde as a grey solid (5.9 g, 59%). 1 H NMR (400 MHz, CDCl3) δ 9.80 (s, 1H), 4.14 (s, 2H).

[0172] Step 4: Synthesis of 2,4-dichloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine.

[0173] To a solution of 2-(2,4,6-trichloropyrimidin-5-yl)acetaldehyde (2.2 g, 9.76 mmol) and aniline (1.09 g, 11.71 mmol) in methanol (60 mL) was added acetic acid (1.0 mL) and sodium cyanoborohydride (1.23 g, 19.52 mmol) at 0 °C. The resulting mixture was stirred between 0 and 20 °C for 17 h. Water (60 mL) was added to the mixture and after 10 min, the resulting precipitate was collected by filtration and dried under vacuum to give 2,4-dichloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine as a white solid (2.0 g, 77%). 1 H NMR (400 MHz, CDCl3) δ 7.69 (dd, J = 8.8Hz, J = 1.2Hz, 2H), 7.39-7.44 (m, 2H), 7.16 (t, J = 7.2Hz, 1H), 4.21 (t, J = 8.8Hz, 2H), 3.17 (t, J = 8.8Hz, 2H); LCMS (ESI) m / z: 266.1 [M+H] + .

[0174] Step 5: Synthesis of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0175] A solution of 2,4-dichloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (100 mg, 0.376 mmol) and morpholine (164 mg, 1.88 mmol) in tetrahydrofuran (10 mL) was heated to 50° C. for 17 h. The mixture was concentrated to dryness, followed by addition of acetonitrile (5 mL) and water (20 mL) to the residue. The resulting precipitate was collected by filtration and dried under vacuum to give 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine as a white solid (67 mg, 56%). 1H NMR (400 MHz, DMSO-d6) δ 7.78 (d, J = 7.6Hz, 2H), 7.39 (t, J = 6.8Hz, J = 2.0Hz, 2H), 7.06 (t, J = 7.2Hz, 1H), 4.10 (t, J = 8.8Hz, 2H), 3.65 (s, 8H), 2.99 (t, J = 8.8Hz, 2H); LCMS (ESI) m / z: 317.1 [M+H]+. Synthesis of 4-(7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 2): [ka]

[0176] Step 1a: Synthesis of morpholine-4-carboximidamide hydrochloride.

[0177] N,N-Diisopropylethylamine (2.58 g, 20.00 mmol) was added to a solution of morpholine (1.74 g, 20.00 mmol) and 1H-pyrazole-1-carboximidamide hydrochloride (2.92 g, 20.00 mmol) in N,N-dimethylformamide (5 mL) at room temperature. The reaction mixture was stirred at room temperature for 16 hours, and ethyl ether (50 mL) was added to the mixture. The oily product at the bottom of the flask solidified with fresh ethyl ether and repeated sonication. The solid was then filtered and dried to give morpholine-4-carboximidamide hydrochloride (3 g, 91%) as a white solid. LCMS (ESI) m / z: 130.1 [M+H] + .

[0178] Step 1: Synthesis of methyl 2-oxotetrahydrofuran-3-carboxylate.

[0179] A solution of dihydrofuran-2(3H)-one (3.36 g, 39.02 mmol) in tetrahydrofuran (5 mL) was added dropwise to lithium hexamethyldisilazide (1.0 M in tetrahydrofuran, 80.0 mL, 80.0 mmol) at -78°C. After stirring at -78°C for 10 min, dimethyl carbonate (3.69 g, 40.98 mmol) was added at the same temperature. The reaction mixture was warmed and stirred at room temperature for 16 h. It was then poured onto a mixture of concentrated hydrochloric acid (15 mL) and ice (150 mL), followed by extraction with ethyl acetate (200 mL x 2). The organic layer was washed with brine, dried over sodium sulfate, and concentrated to give methyl 2-oxotetrahydrofuran-3-carboxylate (4.9 g, 87%). LCMS (ESI) m / z: 144.9 [M+H] + .

[0180] Step 2: Synthesis of 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine.

[0181] Morpholine-4-carboximidamide hydrochloride (575 mg, 3.47 mmol) was added to a solution of methyl 2-oxotetrahydrofuran-3-carboxylate (500 mg, 3.47 mmol) and sodium methoxide (287 mg, 5.31 mmol) in methanol (5 mL) at room temperature. The reaction mixture was refluxed for 2 hours and concentrated. The resulting residue was dissolved in phosphorus oxychloride (5 mL) and heated at 100° C. with stirring for 16 hours. The reaction mixture was then added dropwise to water (100 mL) and then neutralized with 5M aqueous sodium hydroxide solution. It was extracted with ethyl acetate (50 mL×2) and the combined organic layers were washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. The obtained crude product was purified by silica gel column chromatography (n-hexane / ethyl acetate=10 / 1) to obtain 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (236 mg, 23%) as a white solid. LCMS (ESI) m / z: 298.0 [M+H] + .

[0182] Step 3: Synthesis of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0183] A solution of aniline (157 mg, 1.69 mmol) in tetrahydrofuran (3 mL) was added to a solution of sodium hydride (68 mg, 1.70 mmol) in tetrahydrofuran (2 mL) at 0° C. Then the reaction mixture was refluxed for 2 hours and cooled. Then 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (100 mg, 0.34 mmol) was added at room temperature and the resulting mixture was refluxed for 16 hours. It was cooled and then poured into ice water (30 mL) and extracted with ethyl acetate (20 mL×2). The organic layer was washed with brine (20 mL), dried over sodium sulfate and concentrated. The resulting crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate=9 / 1) to give 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (90 mg, 82%). LCMS(ESI)m / z:317.0[M+H] + .

[0184] Step 4: Synthesis of 4-(7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0185] A suspension of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (80 mg, 0.25 mmol) and Pd / C (30 mg) in methanol (10 mL) and ethyl acetate (2 mL) was stirred at room temperature under a hydrogen atmosphere for 30 min. The reaction solution was filtered through Celite and the filtrate was concentrated. The resulting crude product was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 4-(7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (49.2 mg, 70%) as a pale yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.85 (s, 1H), 7.80 (d, J = 8Hz, 2H), 7.37 (t, J = 8Hz, 2H), 7.02 (t, J = 7.6Hz, 1H), 4.04 (t, J = 8.4Hz, 2H), 3.64 (s, 8H), 2.99 (t, J = 8.4Hz, 2H). LCMS (ESI) m / z: 283.1 [M+H] + . Synthesis of 4-(7-(3-fluorophenyl)-4-(pyridin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 3): [ka]

[0186] Step 1: Synthesis of 4-(4-chloro-7-(3-fluorophenyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0187] To a solution of 3-fluoroaniline (181 mg, 1.63 mmol) in THF (20 mL) was added NaH (130 mg, 3.25 mmol) slowly at 0° C. The mixture was stirred at 60° C. for 2 h, then 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (400 mg, 1.35 mmol) was added. The resulting mixture was stirred at 110° C. for 16 h, then quenched with saturated aqueous NH4Cl (20 mL). The mixture was extracted with EtOAc (50×3 mL) and the combined organics were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by SGC (PE / EA=1:1) to give 4-(4-chloro-7-(3-fluorophenyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (270 mg, 59%) as a yellow solid. LCMS (ESI) m / z: 335.0 [M+H] + .

[0188] Step 2: Synthesis of 4-(7-(3-fluorophenyl)-4-(pyridin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0189] To a solution of pyridin-3-ylmethanol (86 mg, 0.79 mmol) in THF (20 mL) was added NaH (32 mg, 0.79 mmol) slowly at 0° C. The mixture was stirred at 0° C. for 2 h, and then 4-(4-chloro-7-(3-fluorophenyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (220 mg, 0.66 mmol) was added. The resulting mixture was stirred at 110° C. for 16 h and concentrated. The resulting crude product was purified by preparative HPLC (0.05% FA / HO:CHCN=5%-95%) to give 4-(7-(3-fluorophenyl)-4-(pyridin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (47.2 mg, 18%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 8.67 (s, 1H), 8.54 (d, J = 4.0Hz, 1H), 7.86 (d, J = 8.0Hz, 1H), 7.77 (d, J = 12.8Hz, 1H), 7.49 (d, J = 8.4Hz, 1H), 7.43 - 7.33 (m, 2H), 6.81-6.76 (m, 1H), 5.45 (s, 2H), 4.03 (t, J = 8.8Hz, 2H), 3.67 (s, 8H), 2.90 (t, J = 8.8Hz, 2H); LCMS (ESI) m / z: 408.1 [M+H]+. Synthesis of 3-(2-morpholino-4-(pyridin-3-ylmethoxy)-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)benzonitrile (compound 4): [ka]

[0190] Step 1: Synthesis of 3-(4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)benzonitrile.

[0191] To a suspension of sodium hydride (40 mg, 1.0 mmol) in tetrahydrofuran (10 mL) was added 3-aminobenzonitrile (48 mg, 0.405 mmol) at 0° C. The reaction mixture was then refluxed for 1 h and cooled to room temperature. A THF solution of 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (100 mg, 0.337 mmol) was added to the mixture, which was then refluxed for another 16 h. After cooling to room temperature, the reaction mixture was quenched with water (50 mL) and the resulting precipitate was collected by filtration, washed with methanol and dried to give 3-(4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)benzonitrile (60 mg, 52%). The crude product was used in the next step without further purification. LCMS (ESI) m / z: 342.0 [M+H] + .

[0192] Step 2: Synthesis of 3-(2-morpholino-4-(pyridin-3-ylmethoxy)-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)benzonitrile.

[0193] Pyridin-3-ylmethanol (48 mg, 0.440 mmol) was added to a suspension of sodium hydride (21 mg, 0.525 mmol) in tetrahydrofuran (10 mL) at room temperature and stirred for 10 minutes. Then 3-(4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)benzonitrile (60 mg, 0.176 mmol) was added. The resulting mixture was refluxed for 12 hours and cooled. Water (30 mL) was added to the mixture and the resulting solid was collected by filtration to give the crude product. This was then purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 3-(2-morpholino-4-(pyridin-3-ylmethoxy)-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)benzonitrile (11.3 mg, 16%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.67 (s, 1H), 8.53 (d, J = 4.8Hz, 1H), 8.16 (d, J = 8Hz, 1H), 8.12 (s, 1H), 7.85 (d, J = 8Hz, 1H), 7.55 (s, 1H), 7.43-7.39 (m, 2H), 5.45 (s, 2H), 4.06 (t, J = 8.6Hz, 2H), 3.67 (s, 8H), 2.92 (t, J = 8.6Hz, 2H). LCMS (ESI) m / z: 415.0 [M+H] + .

[0194] Synthesis of tert-butyl 4-(2-morpholino-4-(pyridin-3-ylmethoxy)-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)piperidine-1-carboxylate (compound 5) and 4-(7-(piperidin-4-yl)-4-(pyridin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 6): [ka]

[0195] Step 1: Synthesis of tert-butyl 4-(6-chloro-5-(2-chloroethyl)-2-morpholinopyrimidin-4-ylamino)piperidine-1-carboxylate.

[0196] To a stirred solution of 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (100 mg, 0.337 mmol) and tert-butyl 4-aminopiperidine-1-carboxylate (135 mg, 0.674 mmol) in acetonitrile (5 mL) was added DIPEA (109 mg, 0.843 mmol) at room temperature and the resulting mixture was refluxed for 16 h. After cooling to room temperature, the mixture was diluted with ethyl acetate (50 mL), washed with water (20 mL), brine (20 mL), dried over sodium sulfate, filtered and concentrated to give tert-butyl 4-(6-chloro-5-(2-chloroethyl)-2-morpholinopyrimidin-4-ylamino)piperidine-1-carboxylate (100 mg, 64%) as a white solid. This material was used in the next step without further purification. LCMS(ESI)m / z:460.1[M+H] + .

[0197] Step 2: Synthesis of tert-butyl 4-(4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)piperidine-1-carboxylate.

[0198] Cesium carbonate (177 mg, 0.543 mmol) was added to a mixture of tert-butyl 4-(6-chloro-5-(2-chloroethyl)-2-morpholinopyrimidin-4-ylamino)piperidine-1-carboxylate (100 mg, 0.217 mmol) and sodium iodide (7 mg, 0.047 mmol) in acetonitrile (10 mL) at room temperature. The resulting mixture was refluxed under nitrogen atmosphere for 4 h. After cooling to room temperature, the mixture was diluted with ethyl acetate (80 mL) and washed with water (30 mL) and brine (30 mL). The organics were dried over sodium sulfate, filtered and concentrated. The resulting crude product was purified by silica gel column chromatography eluting with petroleum ether / ethyl acetate=9 / 1 then 3 / 1 to give tert-butyl 4-(4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)piperidine-1-carboxylate (20 mg, 22%) as a white solid. LCMS (ESI) m / z: 424.3 [M+H] + .

[0199] Step 3: Synthesis of tert-butyl 4-(2-morpholino-4-(pyridin-3-ylmethoxy)-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)piperidine-1-carboxylate.

[0200] A suspension of pyridin-3-ylmethanol (10 mg, 0.092 mmol) and sodium hydride (5 mg, 0.125 mmol) in tetrahydrofuran (3 mL) was stirred at room temperature for 10 minutes, followed by the addition of tert-butyl 4-(4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)piperidine-1-carboxylate (20 mg, 0.047 mmol). The reaction mixture was then refluxed for 72 hours and cooled. It was then diluted with ethyl acetate (80 mL), washed with water (30 mL×2) and brine (20 mL), dried over sodium sulfate, filtered and concentrated. The resulting crude product was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give tert-butyl 4-(2-morpholino-4-(pyridin-3-ylmethoxy)-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)piperidine-1-carboxylate (14.6 mg, 62%) as a white solid. 1 H NMR (400 MHz, MeOD) δ 8.60 (d, J = 1.6Hz, 1H), 8.47 (dd, J = 5.2, 1.6Hz, 1H), 7.90 (dt, J = 8.0, 1.6Hz, 1H), 7.44 (dd, J = 8.0, 0.8Hz, 1H), 5.42 (s, 2H), 4.18 (d, J = 12.4Hz, 2H), 4.03 (pent, J = 7.6Hz, 1H), 3.68 (s, 8H), 3.54 (t, J = 8.4Hz, 2H), 2.82-2.78 (m, 4H), 1.73-1.68 (m, 4H), 1.48 (s, 9H).LCMS (ESI) m / z: 497.1 [M+H] + .

[0201] Step 4: Synthesis of 4-(7-(piperidin-4-yl)-4-(pyridin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0202] Trifluoroacetic acid (1 mL) was added to a solution of tert-butyl 4-(2-morpholino-4-(pyridin-3-ylmethoxy)-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)piperidine-1-carboxylate (60 mg, 0.121 mmol) in dichloromethane (2 mL) at room temperature. After stirring at room temperature for 2 hours, the reaction mixture was concentrated. The residue was subjected to preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 4-(7-(piperidin-4-yl)-4-(pyridin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (9.5 mg, 20%) as a white solid. 1 H NMR (400 MHz, CD3OD) δ 8.60 (s, 1H), 8.48 (d, J = 4.4Hz, 1H), 7.90 (d, J = 8.0Hz, 1H), 7.44 (dd, J = 8.0, 4.8Hz, 1H), 5.42 (s, 2H), 4.08-4.03 (m, 1H), 3.69 (s, 8H), 3.58 (t, J = 8.0Hz, 2H), 3.32-3.30 (m, 2H), 2.91-2.80 (m, 4H), 1.90-1.81 (m, 4H). LCMS (ESI) m / z: 397.1 [M+H] + . Following the protocol described above, the following compounds were synthesized: [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4]

[0203] Synthesis of 4-(7-(pyridin-3-yl)-4-((tetrahydrofuran-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 23): [ka]

[0204] Step 1: Synthesis of 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0205] A solution of pyridin-3-amine (238 mg, 2.53 mmol) in tetrahydrofuran (15 mL) was added to a suspension of sodium hydride (202 mg, 5.06 mmol) in tetrahydrofuran (10 mL) at 0° C. The reaction mixture was then refluxed for 1 h. After cooling to room temperature, 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (500 mg, 1.69 mmol) was added and the mixture was further refluxed for 16 h. The reaction mixture was then poured into ice water (50 mL) and extracted with ethyl acetate (50 mL×2). The organic layer was washed with brine (40 mL), dried over sodium sulfate, filtered and concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate=1 / 3 to 0 / 100) to give 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (400 mg, 74%). LCMS (ESI) m / z: 318.1 [M+H] + .

[0206] Step 2: Synthesis of 4-(7-(pyridin-3-yl)-4-((tetrahydrofuran-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0207] A solution of (tetrahydrofuran-2-yl)methanol (80 mg, 0.78 mmol) in THF (3 mL) was added to a solution of sodium hydride (38 mg, 0.95 mmol) in tetrahydrofuran (5 mL) at 0° C. After stirring at room temperature for 10 minutes, 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (100 mg, 0.31 mmol) was added. The resulting reaction mixture was refluxed for 12 hours. After cooling, the reaction mixture was diluted with ethyl acetate (80 mL), washed with water (30 mL×2) and brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 4-(7-(pyridin-3-yl)-4-((tetrahydrofuran-2-yl)methoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (28.8 mg, 24%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.99 (d, J = 2.4hz, 1H), 8.18-8.15 (m, 2H), 7.36 (dd, J = 8.4, 4.4Hz, 1H), 4.30-4.22 (m, 2H), 4.20-4.13 (m, LCMS (ESI) m / z: 384.1 [M+H] + . Synthesis of 4-(7-phenyl-4-(pyridin-2-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 24): [ka]

[0208] To a solution of pyridin-2-ylmethanol (52 mg, 0.47 mmol) in dry THF (10 mL) was added NaH (28 mg, 0.71 mmol) and the mixture was stirred at 0° C. for 15 min. Then a solution of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (150 mg, 0.47 mmol) in THF (5 mL) was added and the resulting mixture was stirred at 100° C. for an additional 16 h. The reaction was quenched with ice water (20 mL) and extracted with EtOAc (20 mL×3). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative HPLC to give 4-(7-phenyl-4-(pyridin-2-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (28.4 mg, 16%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.54 (dd, J = 4.8, 0.8Hz, 1H), 7.82-7.78 (m, 1H), 7.76(s, 1H), 7.74(s, 1H), 7.42(d, J=8.0Hz, 1H), 7.36-7.29 (m, 3H), 6.97(t, J=7.2Hz, 1H), 5.44(s, 2H), 4.04(t, J=8.8Hz, 2H), 3.59(s, 8H), 2.94 (t, J=8.8Hz, 2H); LCMS (ESI) m / z:390.3 [M+H] + . Following the protocol described above, the following compounds were synthesized: [Table 6-1] [Table 6-2] [Table 6-3]

[0209] Synthesis of tert-butyl 3-(((2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)methyl)pyrrolidine-1-carboxylate (compound 33), 4-(7-(pyridin-3-yl)-4-(pyridin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 34) and 4-(4-((1-methylpyrrolidin-3-yl)methoxy)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 35): [ka]

[0210] Step 1: Synthesis of tert-butyl 3-(((2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)methyl)pyrrolidine-1-carboxylate.

[0211] To a solution of tert-butyl 3-(hydroxymethyl)pyrrolidine-1-carboxylate (84 mg, 0.42 mmol) in THF (15 mL) was carefully added NaH (30 mg, 0.76 mmol) at 0° C. The mixture was stirred at room temperature for 15 min, then 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (120 mg, 0.38 mmol) was added. The resulting mixture was further stirred at 100° C. for 16 h. It was quenched with water (10 mL) and extracted with EA (30×3 mL). The organic layer was washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. The residue was purified by SGC (PE / EA=1:1 to 0:1) to give tert-butyl 3-(((2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)methyl)pyrrolidine-1-carboxylate (135 mg, 74%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 9.05 (d, J = 2.4Hz, 1H), 8.23 ​​(d, J = 4.0Hz, 1H), 8.06 (d, J = 9.2Hz, 1H), 7.28 (s, 1H), 4.35-4.27 (m, 2H), 4.04 (t, J = 8.4Hz, 2H), 3.62 (s, 8H), 3.60-3.34 (m, 3H), 3.22-3.16 (m, 1H), 3.03-2.97 (m, 2H), 2.69-2.64 (m, 1H), 2.10-2.04 (s, 1H), 1.82-1.74 (m, 1H), 1.49 (s, 9H); LCMS (ESI) m / z: 483.3 [M+H]+

[0212] Step 2: Synthesis of 4-(7-(pyridin-3-yl)-4-(pyrrolidin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0213] To a solution of tert-butyl 3-(((2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)methyl)pyrrolidine-1-carboxylate (120 mg, 0.25 mmol) in DCM (5 mL) was added TFA (1 mL) at 0° C. The mixture was stirred at room temperature for 2 h and concentrated. The resulting residue was purified by preparative HPLC (0.05% NH4HCO3 / H2O:CH3CN=5%-95%) to give 4-(7-(pyridin-3-yl)-4-(pyrrolidin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (16.8 mg, 63%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 9.06 (d, J = 2.4Hz, 1H), 8.24 (d, J = 4.4Hz, 1H), 8.10 (d, J = 10.0Hz, 1H), 7.28 (s, 1H), 4.31 (dd, J = 10.8, 6.0Hz, 1H), 4.23 (dd, J = 10.8, 8.0Hz, 1H), 4.04 (t, J = 8.4Hz, 2H), 3.78 (s, 8H), 3.13-3.10 (m, 1H), 3.08-2.93 (m, 4H), 2.82-2.77 (m, 1H), 2.60-2.53 (m, 1H), 2.01-1.95 (m, 1H), 1.60-1.53 ​​(m, 1H); LCMS (ESI) m / z: 383.1 [M+H]+.

[0214] Step 3: Synthesis of 4-(4-((1-methylpyrrolidin-3-yl)methoxy)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0215] To a solution of 4-(7-(pyridin-3-yl)-4-(pyrrolidin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (30 mg, 0.076 mmol) in methanol (5 mL) was added formaldehyde (2.5 mg, 0.083 mmol). The mixture was stirred at room temperature for 2 h, followed by the addition of sodium cyanoborohydride (24 mg, 0.38 mmol) to the mixture. This was then stirred at room temperature for 12 h. The reaction was then quenched with water (5 mL) and extracted with EA (20×3 mL). The organic layer was washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. The residue was purified by preparative HPLC (0.05% NH4HCO3 / H2O:CH3CN=5%-95%) to give 4-(4-((1-methylpyrrolidin-3-yl)methoxy)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (8.7 mg, 28%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 9.06 (d, J = 2.4Hz, 1H), 8.25 (d, J = 3.6Hz, 1H), 8.09 (d, J = 9.6Hz, 1H), 7.28 (s, 1H), 4.33-4.23 (m, 2H), 4.04 (t, J = 8.4Hz, 2H), 3.78 (s, 8H), 3.00 (t, J = 8.4Hz, 2H), 2.85-2.83 (m, 1H), 2.75-2.64 (m, 3H), 2.53-2.49 (m, 1H), 2.45 (s, 3H), 2.13-2.03 (m, 1H), 1.87-1.66 (m, 1H); LCMS (ESI) m / z: 397.2 [M+H]+. Following the protocol described above, the following compounds were synthesized: [Table 7-1] [Table 7-2]

[0216] Synthesis of 2-methyl-1-(4-((1-methylpiperidin-3-yl)methoxy)-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)propan-2-ol (compound 40): [ka]

[0217] Step 1: Synthesis of 1-(6-chloro-5-(2-chloroethyl)-2-morpholinopyrimidin-4-ylamino)-2-methylpropan-2-ol.

[0218] To a stirred solution of 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (200 mg, 0.674 mmol) and 1-amino-2-methylpropan-2-ol (60 mg, 0.674 mmol) in acetonitrile (20 mL) was added N-ethyl-N-isopropylpropan-2-amine (218 mg, 1.687 mmol) at room temperature. The reaction mixture was then refluxed for 48 h. After cooling to room temperature, the mixture was diluted with ethyl acetate (100 mL) and washed with water (30 mL) and brine (30 mL). The organics were dried over sodium sulfate, filtered and concentrated to give 1-(6-chloro-5-(2-chloroethyl)-2-morpholinopyrimidin-4-ylamino)-2-methylpropan-2-ol (200 mg, 85%) as a brown solid. LCMS(ESI)m / z:348.9[M+H] + .

[0219] Step 2: Synthesis of 1-(4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)-2-methylpropan-2-ol.

[0220] Cesium carbonate (466 mg, 1.43 mmol) was added to a solution of 1-(6-chloro-5-(2-chloroethyl)-2-morpholinopyrimidin-4-ylamino)-2-methylpropan-2-ol (200 mg, 0.573 mmol) and sodium iodide (17 mg, 0.113 mmol) in acetonitrile (20 mL) at room temperature. The reaction mixture was refluxed under nitrogen for 4 h, cooled, and then diluted with ethyl acetate (150 mL). The mixture was washed with water (50 mL), brine (30 mL), dried over sodium sulfate, filtered, and concentrated. The resulting crude product was purified by silica gel column chromatography eluting with dichloromethane / methanol=9 / 1 to give 1-(4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)-2-methylpropan-2-ol (100 mg, 55%) as a white solid. LCMS (ESI) m / z: 313.1 [M+H] + .

[0221] Step 3: Synthesis of 2-methyl-1-(4-((1-methylpiperidin-3-yl)methoxy)-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)propan-2-ol.

[0222] A suspension of (1-methylpiperidin-3-yl)methanol (83 mg, 0.64 mmol) and sodium hydride (32 mg, 0.8 mmol) in tetrahydrofuran (10 mL) was stirred at room temperature for 10 minutes, then 1-(4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)-2-methylpropan-2-ol (100 mg, 0.32 mmol) was added. The resulting mixture was refluxed for 48 hours and cooled. It was then diluted with ethyl acetate (80 mL), washed with water (30 mL) and brine (30 mL), dried over sodium sulfate, filtered and concentrated. The crude product was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 2-methyl-1-(4-((1-methylpiperidin-3-yl)methoxy)-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)propan-2-ol (14 mg, 11%) as a pale yellow solid. 1 H NMR (500 MHz, MeOD) δ 6.03 (s, 1H), 4.17-4.07 (m, 2H), 3.73-3.61 (m, 10), 3.22 (s, 2H), 2.93-2.77 (m, 4H), 2.27 (s, 3H), 2.07 (bs, 1H), 1.95-1.89 (m, 1H), 1.76-1.57 (m, 4H), 1.23 (s, 6H), 1.00 (m, 1H). LCMS (ESI) m / z: 406.2 [M+H] + . Synthesis of 4-(4-chloro-7-(3-methylbenzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 41): [ka]

[0223] A mixture of 2,4-dichloro-7-(3-methylbenzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (2.0 g, 6.80 mmol) and morpholine (2.96 g, 34.0 mmol) in tetrahydrofuran (40 mL) was heated to 35° C. for 17 h and concentrated to dryness. MeOH (40 mL) and water (40 mL) were added to the residue and stirred. The resulting precipitate was collected by filtration and dried in vacuum to give 4-(4-chloro-7-(3-methylbenzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (2.0 g, 85%). 1 H NMR (400 MHz, DMSO-d6) δ 7.23 (t, J = 8.0Hz, 2H), 7.05-7.10 (m, 3H), 4.48 (s, 2H), 3.61 (s, 8H), 3.48 (t, J = 8.4Hz, 2H), 2.85 (t, J = 8.4Hz, 2H), 2.29 (s, 3H); LCMS (ESI) m / z: 345.1 [M+H]+. Synthesis of 4-(7-(1-methylpiperidin-4-yl)-4-(pyridin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 42): [ka]

[0224] Step 1: Synthesis of 6-chloro-5-(2-chloroethyl)-N-(1-methylpiperidin-4-yl)-2-morpholinopyrimidin-4-amine.

[0225] To a stirred solution of 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (100 mg, 0.337 mmol) and 1-methylpiperidin-4-amine (38 mg, 0.333 mmol) in acetonitrile (10 mL) was added N-ethyl-N-isopropylpropan-2-amine (109 mg, 0.843 mmol) at room temperature. The reaction mixture was then refluxed for 16 hours and cooled. It was diluted with ethyl acetate (80 mL), washed with water (20 mL), brine (20 mL), dried over sodium sulfate, filtered and concentrated to give 6-chloro-5-(2-chloroethyl)-N-(1-methylpiperidin-4-yl)-2-morpholinopyrimidin-4-amine (100 mg, 79%) as a white solid. LCMS (ESI) m / z: 374.0 [M+H] + .

[0226] Step 2: Synthesis of 4-(4-chloro-7-(1-methylpiperidin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0227] Cesium carbonate (218 mg, 0.669 mmol) was added to a solution of 6-chloro-5-(2-chloroethyl)-N-(1-methylpiperidin-4-yl)-2-morpholinopyrimidin-4-amine (100 mg, 0.267 mmol) and sodium iodide (8 mg, 0.053 mmol) in acetonitrile (20 mL) at room temperature. The resulting mixture was refluxed under nitrogen for 4 hours and cooled. It was diluted with ethyl acetate (150 mL), washed with water (50 mL) and brine (30 mL), dried over sodium sulfate, filtered and concentrated. The resulting crude product was purified by silica gel column chromatography eluting with dichloromethane / methanol=9 / 1 to give 4-(4-chloro-7-(1-methylpiperidin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (30 mg, 0.089 mmol, 33%) as a white solid. LCMS (ESI) m / z: 338.1 [M+H] + .

[0228] Step 3: Synthesis of 4-(7-(1-methylpiperidin-4-yl)-4-(pyridin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0229] To a suspension of sodium hydride (9 mg, 0.225 mmol) in tetrahydrofuran (5 mL) was added pyridin-3-ylmethanol (20 mg, 0.183 mmol) at room temperature and stirred for 10 minutes. Then a solution of 4-(4-chloro-7-(1-methylpiperidin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (30 mg, 0.089 mmol) in THF was added to the mixture and the resulting mixture was refluxed for 48 hours. It was cooled, diluted with ethyl acetate (80 mL), washed with water (30 mL) and brine (30 mL), dried over sodium sulfate, filtered and concentrated. The resulting crude product was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 4-(7-(1-methylpiperidin-4-yl)-4-(pyridin-3-ylmethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (14.5 mg, 40%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 8.61 (d, J = 1.6Hz, 1H), 8.50 (dd, J = 4.8,1.2Hz, 1H), 7.80 (d, J = 8.0Hz, 1H), 7.39 (dd, J = 7.6, 4.8Hz, 1H), 5.35 (s, 2H), 3.74-3.70 (m, 1H), 3.59 (s, 8H), 3.48 (t, J = 8.4Hz, 2H), 2.82-2.79 (m, 2H), 2.71 (t, J = 8.4Hz, 2H), 2.15 (s, 3H), 1.94-1.89 (m, 2H), 1.74-1.68 (m, 2H), 1.56-1.53 ​​(m, 2H). LCMS (ESI) m / z: 411.3 [M+H] + . Synthesis of 3-((2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)propane-1,2-diol (compound 43): [ka]

[0230] Step 1: Synthesis of 4-(4-((2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0231] A solution of (2,2-dimethyl-1,3-dioxolan-4-yl)methanol (90 mg, 0.68 mmol) in THF (5 mL) was added to a suspension of sodium hydride (27 mg, 0.68 mmol) in THF (5 mL) at 0° C. The reaction mixture was refluxed for 2 h and cooled. A solution of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (100 mg, 0.34 mmol) in 3 mL of THF was then added and the resulting mixture was stirred at reflux for 16 h. The reaction mixture was then diluted with ethyl acetate (30 mL) and the resulting organic medium was washed with brine (10 mL), dried over sodium sulfate and concentrated to give 100 mg of the target compound, which was used in the next step without further purification.

[0232] Step 2: Synthesis of 3-((2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)propane-1,2-diol.

[0233] A solution of 4-(4-((2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (100 mg, 0.24 mmol) in water (1 mL) and acetic acid (5 mL) was stirred overnight at 80° C. The resulting mixture was concentrated and the resulting crude product was purified by preparative HPLC to give 5.2 mg of the title compound as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 7.76 (d, J = 8.0Hz, 2H), 7.34 (t, J = 7.6Hz, 2H), 6.97 (t, J = 7.6Hz, 1H), 4.87 (d, J = 5.2Hz 1H), 4.64 (t, J = 8.8Hz, 1H), 4.25~4.30 (m, 1H), 4.00~4.15 (m, 1H), 4.02 (t, J = 8.4Hz, 2H), 3.75~3.80 (m, 1H), 3.67 (s, 8H), 3.40(t, J = 5.6Hz, 2H), 2.88(t, J = 9.2Hz, 2H); LCMS (ESI) m / z: 373.0 [M+H]+. Synthesis of 4-(7-phenyl-4-(2-(pyridin-2-yl)ethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 44): [ka]

[0234] Step 1: Synthesis of 4-(4-chloro-5-(2-chloroethyl)-6-(2-(pyridin-2-yl)ethoxy)pyrimidin-2-yl)morpholine.

[0235] A solution of 2-(pyridin-2-yl)ethanol (830 mg, 6.74 mmol) in DMF was added to a solution of sodium hydride (270 mg, 6.74 mmol) in DMF (60 mL) at 0° C. The resulting mixture was warmed and stirred at room temperature for 10 min, followed by the addition of 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (2 g, 6.74 mmol). The reaction mixture was further stirred at room temperature for 48 h. It was quenched with water (200 mL) and extracted with ethyl acetate (300 mL×2). The combined organic layers were washed with water (200 mL×2), brine (200 mL), dried over sodium sulfate, filtered and concentrated. The resulting crude product was purified by silica gel column chromatography eluting with petroleum ether / ethyl acetate=3 / 1 to give 4-(4-chloro-5-(2-chloroethyl)-6-(2-(pyridin-2-yl)ethoxy)pyrimidin-2-yl)morpholine (1.9 g, 74%) as an off-white solid. LCMS (ESI) m / z: 398.1 [M+16] + .

[0236] Step 2: Synthesis of 4-(7-phenyl-4-(2-(pyridin-2-yl)ethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0237] A mixture of 4-(4-chloro-5-(2-chloroethyl)-6-(2-(pyridin-2-yl)ethoxy)pyrimidin-2-yl)morpholine (100 mg, 0.261 mmol), aniline (49 mg, 0.526 mmol), tris(dibenzylideneacetone)dipalladium (24 mg, 0.026 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (30 mg, 0.052 mmol) and cesium carbonate (170 mg, 0.522 mmol) in dioxane (5 mL) was stirred at 100° C. for 16 h under nitrogen atmosphere. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (100 mL), washed with water (30 mL×2), brine (30 mL), dried over sodium sulfate, filtered and concentrated. The resulting residue was subjected to silica gel column chromatography (eluted with petroleum ether / ethyl acetate=2 / 1) followed by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, and the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate solution) to give 4-(7-phenyl-4-(2-(pyridin-2-yl)ethoxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (53.8 mg, 51%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.50 (dd, J = 4.8, 0.8Hz, 1H), 7.75-7.70 (m, 2H), 7.35-7.31 (m, 3H), 7.25-7.22 (m, 1H), 6.96 (t, J = 7.2Hz, LCMS (ESI) m / z: 404.2 [M+H] + .

[0238] Synthesis of 4-(4-methyl-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 45) and 2-methyl-1-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)propan-2-ol (compound 46): [ka]

[0239] Step 1: Synthesis of 4-(4-methyl-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0240] A mixture of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 0.158 mmol), 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (40 mg, 0.316 mmol), tris(dibenzylideneacetone)dipalladium (15 mg, 0.016 mmol), tris(dibenzylideneacetone)dipalladium (9 mg, 0.032 mmol) and cesium carbonate (103 mg, 0.316 mmol) in dimethyl sulfoxide (2 mL) and water (0.5 mL) was stirred at 140 °C for 16 h under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (80 mL), washed with water (40 mL×3), brine (30 mL), dried over sodium sulfate, filtered and concentrated. The crude product obtained was purified by silica gel column chromatography eluting with petroleum ether / ethyl acetate=6 / 1 to give 4-(4-methyl-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (31.6 mg, 68%). 1H NMR (500 MHz, DMSO-d6) δ 7.80 (d, J = 7.5Hz, 2H), 7.36 (dd, J = 8.5, 7.5Hz, 2H), 7.00 (t, J = 7.5Hz, 1H), 4.03 (t, J = 8.5Hz, 2H), 3.64 (s, 8H), 2.95 (t, J = 8.5Hz, 2H), 2.13 (s, 3H). LCMS (ESI) m / z: 297.2 [M+H] + .

[0241] Step 2: Synthesis of 2-methyl-1-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)propan-2-ol.

[0242] To a stirred solution of 4-(4-methyl-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (100 mg, 0.338 mmol) in tetrahydrofuran (5 mL) was added n-butyllithium (0.25 mL, 0.506 mmol) at 0° C., and the resulting mixture was stirred at 0° C. for 0.5 h. Then propan-2-one (29 mg, 0.101 mmol) was added, and the mixture was further stirred at room temperature for 2 h. Then water (20 mL) was added, and the mixture was extracted with ethyl acetate (30 mL×3). The organic layer was dried over sodium sulfate, filtered, and concentrated. The resulting crude product was purified by preparative HPLC (column Xbridge 21.2×250 mm C18, 10 um, mobile phase A: water (10 mmol / L ammonium bicarbonate) B: acetonitrile) to give 2-methyl-1-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)propan-2-ol (35.7 mg, 30%). 1H NMR (400 MHz, DMSO-d6) δ 7.81 (d, J = 8.4Hz, 2H), 7.37 (t, J = 7.6Hz, 2H), 7.01(t, J = 6.4Hz, 1H), 5.01 (s, 1H), 4.03 (t, J = 8Hz, 2H), 3.66-3.60 (m, 8H), 3.00 (t, J - 8.4Hz, 2H), 2.53 (s, 2H), 1.17 (s, 6H); LC-MS: m / z=355.2(M+H) + . Synthesis of 2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine-4-carbonitrile (compound 47): [ka]

[0243] A mixture of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (100 mg, 0.316 mmol), zinc cyanide (74 mg, 0.631 mmol) and bis(tri-tert-butylphosphine)palladium(0) (32 mg, 0.063 mmol) in N,N-dimethylacetamide (4 mL) was heated at 150° C. for 0.5 h under nitrogen atmosphere in a sealed vial with microwave irradiation. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (80 mL) and washed with water (40 mL×3) and brine (30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The resulting crude product was purified by silica gel column chromatography eluting with petroleum ether / ethyl acetate=6 / 1 to give 2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine-4-carbonitrile (33.0 mg, 34%) as a yellow solid. 1H NMR (400 MHz, DMSO-d6) δ 7.81 (d, J = 8.0Hz, 2H), 7.42 (t, J = 8Hz, 2H), 7.12 (s, 1H), 4.16 (t, J = 8.0Hz, 2H), 3.65 (s, 8H), 3.16 (t, J = 8.0Hz, 2H). LCMS (ESI) m / z: 308.1 [M+H] + . Synthesis of 4-(4-methoxy-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 48): [ka]

[0244] To a solution of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (100 mg, 0.34 mmol) in methanol (80 mL) was added sodium methoxide (8 mL). The mixture was refluxed overnight and concentrated. The resulting crude product was purified by preparative HPLC to give 4-(4-methoxy-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (18.3 mg) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.75 (d, J = 6.8Hz, 2H), 7.34 (t, J = 6.0Hz, 2H), 6.97 (t, J = 5.6Hz, 1H), 4.01 (t, J = 6.8Hz, 2H), 3.85 (s, 3H), 3.67 (s, 8H), 2.86 (t, J = 6.8Hz, 2H); LCMS (ESI) m / z: 313 [M+H]+.

[0245] Synthesis of 4-(7-phenyl-4-(pyridin-2-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 49) and 1-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)pyridin-2(1H)-one (compound 50) [ka]

[0246] To a solution of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (100 mg, 0.32 mmol) in DMF (10 mL) was added pyridin-2-ol (33 mg, 0.35 mmol) and K2CO3 (88 mg, 0.64 mmol) and the resulting mixture was stirred at 140° C. for 16 h. The reaction was then quenched with water (5 mL) and extracted with EtOAc (20×3 mL). The organic layers were combined, washed with brine (30 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC (0.05% NH4HCO3 / H2O:CH3CN=5%-95%) to give 4-(7-phenyl-4-(pyridin-2-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (9.3 mg, 8%) and 1-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)pyridin-2(1H)-one (9.0 mg, 8%) as a yellow solid.

[0248] Compound 49: 1 H NMR (400 MHz, CDCl3) δ 8.31 (dd, J = 4.8, 1.2Hz, 1H), 7.79-7.75 (m, 3H), 7.41-7.36 (m, 2H), 7.15-7.04 (m, 3H), 4.07 (t, J = 8.4Hz, 2H), 3.74-3.69 (m, 8H), 2.92 (t, J=8.4Hz, 2H); LCMS (ESI) m / z: 376.1 [M+H]+.

[0249] Compound 50: 1 H NMR (400 MHz, CDCl3) δ 7.77 (d, J =7.6Hz, 2H), 7.67 (dd, J =1.6, 6.4Hz, 1H), 7.43-7.39 (m, 3H), 7.10 (t, J =7.6Hz, 1H), 7.63 (d, J =9.2Hz, LCMS (ESI) m / z: 376.1 [M+H]+. Following the protocol described above, the following compounds were synthesized: [Table 8-1] [Table 8-2]

[0247] Synthesis of 4-(7-phenyl-4-(pyridin-2-ylmethyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 57): [ka]

[0248] A solution of 2-methylpyridine (64 mg, 0.7 mmol) in tetrahydrofuran (15 mL) was added to n-BuLi (1 mL, 2.5 mmol, 2.5 M solution in hexanes) at 0° C. and stirred for 1 h. A solution of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (200 mg, 0.64 mmol) in THF was then added and the resulting mixture was allowed to warm to room temperature and stirred for 16 h. The reaction was then quenched with saturated aqueous NH4Cl (10 mL) and extracted with EtOAc (15×3 mL). The organic layers were combined, washed with brine (30 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC (0.05% FA / H2O:CH3CN=5%-95%) to give 4-(7-phenyl-4-(pyridin-2-ylmethyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (39.3 mg, 17%) as a white solid.

[0252] 1 H NMR (400 MHz, DMSO-d6) δ 8.47 (d, J = 4.0Hz, 1H), 7.80 (d, J = 8.0Hz, 2H), 7.74-7.00 (m, 1H), 7.39 - 7.33 (m, 3H), 7.25-7.22 (m, 1H), 7.01 (t, J = 7.2Hz, 1H), 4.02 (t, J = 8.4Hz, 2H), 3.95 (s, 2H), 3.63 (s, 8H), 2.92 (t, J = 8.4Hz, 2H); LCMS (ESI) m / z: 374.3 [M+H]+. Following the protocol described above, the following compounds were synthesized: [Table 9]

[0249] Synthesis of 4-(7-phenyl-4-((pyridin-3-yloxy)methyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 59) and 5-hydroxy-1-((2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)methyl)pyridin-1-ium-3-ylium (compound 60): [ka]

[0250] Step 1: Synthesis of methyl 2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine-4-carboxylate.

[0251] A solution of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (500 mg, 1.578 mmol), triethylamine (479 mg, 4.734 mmol), palladium(II) acetate (36 mg, 0.160 mmol) and 1,1'-bis(diphenylphosphino)ferrocene (131 mg, 0.236 mmol) in methanol (12 mL) and dimethylsulfoxide (15 mL) was stirred at 80° C. for 16 h under carbon monoxide atmosphere. After cooling to room temperature, the reaction mixture was filtered through Celite and the filtrate was diluted with ethyl acetate (150 mL) and washed with water (40 mL×3) and brine (30 mL). The organics were dried over sodium sulfate, filtered, concentrated, and the resulting crude product was purified by silica gel column chromatography eluting with PE / EA=3 / 1 to give methyl 2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine-4-carboxylate (400 mg, 74%) as a yellow solid. LCMS (ESI) m / z: 341.1 [M+H] + .

[0252] Step 2: Synthesis of (2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)methanol.

[0253] Lithium aluminum hydride (1.76 mL, 1.76 mmol) was added portionwise to a solution of methyl 2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine-4-carboxylate (400 mg, 1.175 mmol) in tetrahydrofuran (20 mL) at 0° C. The solution was stirred at 0° C. for 1 h, then quenched with sodium sulfate decahydrate (2 g), filtered through Celite, and washed with dichloromethane. The filtrate was concentrated to give (2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)methanol (170 mg, 46%) as a white solid. LCMS (ESI) m / z: 313.1 [M+H] + This crude product was used in the next step without further purification. Step 3: Synthesis of Compound 59 and Compound 60:

[0254] To a solution of triphenylphosphine (118 mg, 0.450 mmol), pyridin-3-ol (43 mg, 0.452 mmol) and (2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)methanol (70 mg, 0.224 mmol) in tetrahydrofuran (15 mL) was added DIAD (91 mg, 0.450 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 hour and concentrated. The residue was subjected to preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give compound 59 (17.7 mg, 20%) and compound 60 (12 mg, 14%) as white solids.

[0259] Compound 59: 1H NMR (400 MHz, DMSO-d6) δ 8.36 (d, J = 2.8Hz, 1H), 8.20-8.18 (m, 1H), 7.81 (d, J = 8Hz, 2H), 7.45-7.32 (m, 4H), 7.03 (t, J = 7.6Hz, 1H), 5.03 (s, 2H), 4.05 (t, J = 8.4Hz, 2H), 3.65 (s, 8H), 3.05 (t, J = 8.4Hz, 2H). LCMS (ESI) m / z: 390.1 [M+H] + .

[0260] Compound 60: 1 H NMR (400 MHz, DMSO-d6) δ 7.79 (d, J = 6.0Hz, 2H), 7.46-7.43 (m, 2H), 7.38 (t, J = 6.0Hz, 2H), 7.29 (dd, J = 7.2, 4.4Hz, 1H), 7.05 (t, J = LCMS (ESI) m / z: 390.1 [M]. Synthesis of 4-(7-phenyl-4-(((tetrahydro-2H-pyran-4-yl)oxy)methyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 61): [ka]

[0255] Step 1: Synthesis of (2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)methyl methanesulfonate.

[0256] Methanesulfonyl chloride (37 mg, 0.33 mmol) was added to a solution of (2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)methanol (70 mg, 0.22 mmol) and triethylamine (44 mg, 0.44 mmol) in dichloromethane (8 mL) at 0° C. The reaction mixture was stirred at 0° C. under nitrogen atmosphere for 1 h, then quenched with saturated aqueous sodium bicarbonate (10 mL) and extracted with dichloromethane (20 mL×3). The organic layer was washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated to give (2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)methyl methanesulfonate (70 mg, 72%) as a brown solid. The crude product was used in the next step without further purification. LCMS(ESI)m / z:391.0[M+H] + .

[0257] Step 2: Synthesis of 4-(7-phenyl-4-(((tetrahydro-2H-pyran-4-yl)oxy)methyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0258] A suspension of tetrahydro-2H-pyran-4-ol (27 mg, 0.27 mmol) and sodium hydride (11 mg, 0.27 mmol) in tetrahydrofuran (10 mL) was stirred at room temperature for 30 minutes, followed by the addition of (2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)methyl methanesulfonate (70 mg, 0.18 mmol) to the mixture. The resulting mixture was then stirred at 80° C. for 16 hours, then quenched with water (10 mL) and extracted with dichloromethane (20×3 mL). The organic layer was washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated. The residue was subjected to preparative HPLC (0.05% NH4HCO3 / H2O:CH3CN=5%-95%) to give 4-(7-phenyl-4-(((tetrahydro-2H-pyran-4-yl)oxy)methyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (4.3 mg, 6%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.79 (d, J = 7.6Hz, 2H), 7.41-7.37 (m, 2H), 7.07 (t, J = 7.6Hz, 1H), 4.46 (s, 2H), 4.06 (t, J = 8.4Hz, 2H), 4.01-3.96 (m, 2H), 3.78 (s, 8H), 3.69 - 3.62 (m, 1H), 3.53-3.47 (m, 2H), 3.17 (t, J = 8.4hz, 2H), 2.01-1.97 (m, 2H), 1.71-1.67 (m, 2H); LCMS (ESI) m / z: 397.2 [M+H]+. Synthesis of 4-(7-phenyl-4-((pyridin-2-ylmethoxy)methyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 62): [ka]

[0259] To a solution of pyridin-2-ylmethanol (29 mg, 0.27 mmol) in THF (8 mL) was added sodium hydride (11 mg, 0.27 mmol) portionwise at 0° C. The mixture was stirred at 0° C. for 30 min, followed by the addition of (2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)methyl methanesulfonate (70 mg, 0.18 mmol). The resulting mixture was stirred at 80° C. for 16 h, then quenched with water (10 mL) and extracted with dichloromethane (20 mL×3). The organic layer was washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by preparative HPLC (0.05% NH4HCO3 / H2O:CH3CN=5%-95%) to give 4-(7-phenyl-4-((pyridin-2-ylmethoxy)methyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (10.8 mg, 15%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 8.59 (d, J = 4.4Hz, 1H), 7.79 (d, J = 8.0Hz, 2H), 7.74 (dt, J = 8.0, 2.0Hz, 1H), 7.53 (d, J = 8.0Hz, 1H), 7.40 (t, J = 7.6Hz, 2H), 7.25-7.22 (m, 1H), 7.06 (t, J = 7.6Hz, 1H), 4.77 (s, 2H), 4.56 (s, 2H), 4.06 (t, J = 8.4Hz, 2H), 3.79 (s, 8H), 3.16 (t, J = 8.4Hz, 2H); LCMS (ESI) m / z: 404.1 [M+H]+. Following the protocol described above, the following compounds were synthesized: [Table 10]

[0260] Synthesis of 4-(7-(5-phenylpyridin-3-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 67): [ka]

[0261] Step 1: Synthesis of 4-(4-chloro-5-(2-chloroethyl)-6-(pyridin-3-yloxy)pyrimidin-2-yl)morpholine.

[0262] To a solution of pyridin-3-ol (122 mg, 1.29 mmol) in dry N,N-dimethylacetamide (8 mL) was added sodium hydride (100 mg, 2.5 mmol) portionwise at 0° C. The mixture was stirred at room temperature for 10 min, followed by the addition of 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (380 mg, 1.29 mmol) and the mixture was further stirred at room temperature for 2 h. The reaction was quenched by adding water and extracted with ethyl acetate (20 mL×3) and then washed with water (20 mL). The organic layer was dried and concentrated. The residue was subjected to flash chromatography eluting with 0-50% ethyl acetate in petroleum ether to give 4-(4-chloro-5-(2-chloroethyl)-6-(pyridin-3-yloxy)pyrimidin-2-yl)morpholine as a yellow solid (30 mg, 66%). LCMS(ESI)m / z:354.9[M+H] + .

[0263] Step 2: Synthesis of 4-(7-(5-phenylpyridin-3-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0264] To a mixture of 5-phenylpyridin-3-amine (34 mg, 0.2 mmol), 4-(4-chloro-5-(2-chloroethyl)-6-(pyridin-3-yloxy)pyrimidin-2-yl)morpholine (35 mg, 0.1 mmol) in dioxane (10 mL) was added cesium carbonate (98 mg, 0.3 mmol), tris(dibenzylideneacetone)dipalladium(0) (0.01 mol, 9 mg) and Xantphos (12 mg, 0.02 mmol). The resulting mixture was stirred at 100° C. for 16 h under argon atmosphere. Ethyl acetate (40 mL) was added to the mixture, which was washed with water (10 mL×2). The organic layer was dried and concentrated, and the residue was subjected to preparative HPLC (BOSTON pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile / 0.1% formic acid) to give 4-(7-(5-phenylpyridin-3-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine as a yellow solid (25mg, 55%). 1 H NMR (400 MHz, DMSO-d6) δ 8.98 (d, J = 2.0Hz, 1H), 8.59-8.55 (m, 2H), 8.49 (d, J = 2.4Hz, 1H), 8.43 (dd, J = 4.8Hz, 1H), 7.75-7.67 (m, LCMS (ESI) m / z: 453.0 [M+H] + . Synthesis of 4-(7-(2-phenyl-2H-1,2,3-triazol-4-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 68): [ka]

[0265] Step 1: Synthesis of 4-nitro-2-phenyl-2H-1,2,3-triazole.

[0266] To a solution of 4-nitro-2H-1,2,3-triazole (228 mg, 2 mmol) and phenylboronic acid (488 mg, 4 mmol) in dichloromethane (10 mL) was added cupric acetate (543 mg, 3 mmol) and pyridine (632 mg, 8 mmol). The reaction mixture was stirred at room temperature under oxygen atmosphere for 3 h. The mixture was concentrated and the crude product was purified by column chromatography eluting with 15% acetate in petroleum ether to give 4-nitro-2-phenyl-2H-1,2,3-triazole (isomer A) as a yellow solid (230 mg, 60%). 1 H NMR (400 MHz, CDCl3) δ 8.37 (s, 1H), 8.19-8.08 (m, 2H), 7.61-7.46 (m, 3H); LCMS (ESI) m / z: 191.0 [M+H] + . Note: Regioisomer B was used in the synthesis of compound 69.

[0267] Step 2: Synthesis of 2-phenyl-2H-1,2,3-triazol-4-amine.

[0268] To a solution of 4-nitro-2-phenyl-2H-1,2,3-triazole (230 mg, 1.2 mmol) in methanol (6 mL) was added palladium (10% on activated carbon, 30 mg). The mixture was stirred at room temperature under hydrogen atmosphere for 3 h. It was filtered and the filtrate was concentrated to give 2-phenyl-2H-1,2,3-triazol-4-amine as a white solid (180 mg, 94%). LCMS (ESI) m / z: 161.1 [M+H] + .

[0269] Step 3: Synthesis of 4-(7-(2-phenyl-2H-1,2,3-triazol-4-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0270] To a mixture of 2-phenyl-2H-1,2,3-triazol-4-amine (32 mg, 0.2 mmol) and 4-(4-chloro-5-(2-chloroethyl)-6-(pyridin-3-yloxy)pyrimidin-2-yl)morpholine (35 mg, 0.1 mmol) in dioxane (10 mL) was added cesium carbonate (100 mg, 0.3 mmol), tris(dibenzylideneacetone)dipalladium(0) (0.01 mmol, 9 mg) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (12 mg, 0.02 mmol). The resulting mixture was stirred at 100° C. under argon atmosphere for 16 h. The reaction mixture was then diluted with water (30 mL) and extracted with ethyl acetate (20 mL×3). The combined organic phase was dried and concentrated. The resulting crude product was purified by preparative HPLC (BOSTON pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile / 0.1% formic acid) to give 4-(7-(2-phenyl-2H-1,2,3-triazol-4-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine as a white solid (18mg, 20.4%). 1 H NMR (400 MHz, DMSO) δ 8.49 (d, J = 2.7Hz, 1H), 8.45 (s, 1H), 8.44 (d, J = 4.7Hz, 1H), 7.96 (d, J = 7.6Hz, 2H), 7.70-7.67 (m, 1H), 7.56 (t, J = 8.0Hz, 2H), 7.47 (d, J = 8.3Hz, 1H), 7.38 (t, J = 7.4Hz, 1H), 4.21 (t, J = 8.5Hz, 2H), 3.60 (bs, 4H), 3.53 (bs, 4H), 3.08 (t, J = 8.0Hz, 2H); LCMS (ESI) m / z: 443.0 [M+H] + .

[0271] Using similar protocols as described above, the following compounds were synthesized: [Table 11]

[0272] Synthesis of 4-(7-(1-phenyl-1H-pyrazol-4-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 71): [ka]

[0273] To a mixture of 1-phenyl-1H-pyrazol-4-amine (34 mg, 0.2 mmol) and 4-(4-chloro-5-(2-chloroethyl)-6-(pyridin-3-yloxy)pyrimidin-2-yl)morpholine (35 mg, 0.1 mmol) in dioxane (10 mL), cesium carbonate (98 mg, 0.3 mmol), tris(dibenzylideneacetone)dipalladium(0) (0.01 mol, 9 mg) and Xantphos (12 mg, 0.02 mmol) were added. The resulting mixture was stirred at 100° C. for 16 h under argon atmosphere. The mixture was then extracted with ethyl acetate (20 mL×2) and washed with water (10 mL×2). The organic layer was dried and concentrated, and the resulting residue was subjected to preparative HPLC (BOSTON pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile / 0.1% formic acid) to give 4-(7-(1-phenyl-1H-pyrazol-4-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (20mg, 45%). 1H NMR (400 MHz, DMSO-d6) δ 8.48 (s, 1H), 8.47 (d, J = 2.8Hz, 1H), 8.41 (dd, J = 4.8, 1.2Hz, 1H), 8.26 (s, 1H), 7.82 (d, J = 8.4Hz, 2H), 7.67-7.64 (m, 1H), 7.50 (t, J = 8.4Hz, 2H), 7.47-7.44 (m, 1H), 7.30 (t, J = 7.6Hz, 1H), 4.01 (t, J = 8.4Hz, 2H), 3.61 - 3.59 (m, 4H), 3.52 - 3.48 (m, 4H), 3.05 (t, J = 8.4Hz, 2H). LCMS (ESI) m / z: 442.2 [M+H] + . Synthesis of (2-morpholino-4-(pyridin-3-yloxy)-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)(phenyl)methanone (compound 72): [ka]

[0274] Step 1: Synthesis of (4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)(phenyl)methanone.

[0275] A mixture of 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (600 mg, 2.0 mmol), benzamide (242 mg, 2.0 mmol), tris(dibenzylideneacetone)dipalladium(0) (92 mg, 0.1 mmol), Xantphos (116 mg, 0.2 mmol) and cesium carbonate (1.3 g, 4.0 mmol) in dioxane (20 mL) was stirred at 100° C. under nitrogen atmosphere for 4 h. The mixture was poured into water and extracted with ethyl acetate (150 mL×2). The combined organic phase was concentrated and the resulting residue was subjected to silica gel column chromatography (50% ethyl acetate in petroleum ether) to give (4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)(phenyl)methanone (320 mg, 82%) as a white solid. LCMS (ESI) m / z: 345.1 / 347.1 [M+H] + .

[0276] Step 2: Synthesis of (2-morpholino-4-(pyridin-3-yloxy)-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)(phenyl)methanone.

[0277] A mixture of (4-chloro-2-morpholino-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)(phenyl)methanone (280 mg, 0.81 mmol), pyridin-3-ol (77 mg, 0.81 mmol) and cesium carbonate (527 mg, 1.62 mmol) in N,N-dimethylacetamide (10 mL) was stirred at 120° C. for 16 h. The resulting precipitate was filtered off and the filtrate was purified by preparative HPLC (column Xbridge 21.2×250 mm C18, 10 um, mobile phase A: water (10 mmol / L ammonium bicarbonate) B: acetonitrile) to give (2-morpholino-4-(pyridin-3-yloxy)-5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)(phenyl)methanone (10 mg) and 4-(4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (80 mg, hydrolyzed by-product—which was later converted to the desired product by amidation reaction using benzoyl chloride) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 8.49 (d, J = 2.5Hz, 1H), 8.44 (dd, J = 4.7, 1.2Hz, 1H), 7.55 - 7.50 (m, 2H), 7.48 (ddd, J = 8.3, 2.7, 1.4Hz, 1H), 7.43 - 7.29 (m, 4H), 4.26 (t, J = 4.0Hz, 2H), 3.39 (bs, 4H), 3.24 - 2.76 (m, 6H); LCMS (ESI) m / z: 404.1 [M+H] + . Synthesis of 1-(2-morpholino-4-(pyridin-4-yl)-5,6-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-phenylethan-1-one (compound 73): [ka]

[0278] Step 1: Synthesis of 4-(7-(4-methoxybenzyl)-4-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0279] To a solution of 4-(4-chloro-7-(4-methoxybenzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (300 mg, 0.833 mmol) in DMSO (10 mL) was added pyridin-4-ylboronic acid (123 mg, 1.0 mmol), tris(dibenzylideneacetone)dipalladium (60 mg, 0.631 mmol), tricyclohexylphosphane (60 mg, 0.631 mmol) and cesium carbonate (541 mg, 1.66 mmol). The resulting mixture was stirred at 130° C. for 8 h and water (5 mL) was added. The mixture was extracted with ethyl acetate (20×3 mL) and the organic layers were combined, washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. The residue was subjected to flash chromatography on silica gel (petroleum ether:ethyl acetate=65:35) to give the target product as a yellow solid (330 mg, 98.0%).

[0280] Step 2: Synthesis of 4-(4-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0281] To a solution of 4-(7-(4-methoxybenzyl)-4-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (330 mg, 0.819 mmol) in TFA (10 mL) was added concentrated sulfuric acid (5 mL). The mixture was stirred at 90° C. for 2 h and then diluted with water (20 mL). The mixture was extracted with ethyl acetate (20 mL×3), the organic layers were combined, washed with brine (30 mL), dried over sodium sulfate, filtered, and concentrated. The residue was subjected to flash chromatography on silica gel (petroleum ether:ethyl acetate=45:55) to give the target product as a yellow solid (220 mg, 94.56%).

[0282] Step 3: Synthesis of 1-(2-morpholino-4-(pyridin-4-yl)-5,6-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-phenylethan-1-one.

[0283] To a solution of 4-(4-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (80 mg, 0.283 mmol) in acetonitrile (10 mL) was added 2-phenylacetyl chloride (520 mg, 3.39 mmol), pyridine (44 mg, 0.566 mmol) and N,N-dimethylpyridin-4-amine (69 mg, 0.566 mmol). The mixture was stirred at 30° C. for 8 h, then quenched with water (5 mL) and extracted with ethyl acetate (20 mL×3). The organic layers were combined, washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. The residue was subjected to preparative HPLC (0.05% NH4HCO3 / H2O:CH3CN=5%-95%) to give 1-(2-morpholino-4-(pyridin-4-yl)-5,6-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-phenylethan-1-one (65.4 mg, 57.6%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.74 (d, J = 6.0Hz, 2H), 7.87 (dd, J = 4.6, 1.5Hz, 2H), 7.33 - 7.23 (m, 5H), 4.49 (s, 2H), 4.08 - 3.95 (m, 2H), 3.81 - 3.58 (m, 8H), 3.27 - 3.18 (m, 2H); LCMS (ESI) m / z: 402.1 [M+H]+. Synthesis of 4-(7-(2-phenylpyrimidin-4-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 74): [ka]

[0284] Step 1: Synthesis of 2-phenylpyrimidin-4-amine.

[0285] A mixture of 2-chloropyrimidin-4-amine (1.17 g, 10 mmol), phenylboronic acid (1.83 g, 15 mmol), bis(triphenylphosphino)dichloropalladium(II) (700 mg, 1 mmol) and sodium carbonate (3.18 g, 30 mmol) in dioxane (50 mL) and water (5 mL) was stirred at 90 °C under nitrogen for 16 h. The mixture was concentrated and purified by flash chromatography eluting with 0-50% ethyl acetate in petroleum ether to give 2-phenylpyrimidin-4-amine as a yellow solid (1.5 g, 88%). LCMS (ESI) m / z: 172.2 [M+H] + .

[0286] Step 2: Synthesis of 4-(4-chloro-7-(2-phenylpyrimidin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0287] To a solution of 2-phenylpyrimidin-4-amine (188 mg, 1.1 mmol) in tetrahydrofuran (10 mL) was slowly added sodium hydride (80 mg, 2 mmol) at 0° C. The mixture was stirred at room temperature for 5 min and at 80° C. for 2 h, followed by the addition of 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (295 mg, 1 mmol). The resulting mixture was further stirred at 80° C. for 16 h. The reaction was quenched by the addition of water (10 mL) and ethyl acetate (20 mL). The resulting precipitate was collected by filtration and dried to give 4-(4-chloro-7-(2-phenylpyrimidin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine as a white solid (100 mg, 25%). LCMS(ESI)m / z:395.0[M+H] + .

[0288] Step 3: Synthesis of 4-(7-(2-phenylpyrimidin-4-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0289] A mixture of 4-(4-chloro-7-(2-phenylpyrimidin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (40 mg, 0.11 mmol), pyridin-3-ol (19 mg, 0.2 mmol) and cesium carbonate (98 mmol, 0.3 mg) in N,N-dimethylacetamide (2 mL) was stirred at 150° C. for 1 h under microwave irradiation. The resulting mixture was filtered and the filtrate was purified by preparative HPLC (BOSTON pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile / 0.1% ammonium bicarbonate) to give 4-(7-(2-phenylpyrimidin-4-yl)-4-(pyridin-3-yloxy)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine as a yellow solid (30mg, 66%). 1 H NMR (400 MHz, DMSO-d6) δ 8.67 (d, J = 6.0Hz, 1H), 8.51 (d, J = 2.8Hz, 1H), 8.46-8.39 (m, 4H), 7.73-7.69 (m, 1H), 7.54-7.47 (m, 4H), 4.45 (t, J = 8.0Hz, 2H), 3.62 - 3.51 (m, 8H), 3.07 (t, J = 8.0Hz, 2H). LCMS (ESI) m / z: 454.2 [M+H] + . Synthesis of 4-(4-((5-phenylpyridin-3-yl)oxy)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 75): [ka]

[0290] Step 1: Synthesis of 4-(4-((5-bromopyridin-3-yl)oxy)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0291] To a solution of 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (100 mg, 0.315 mmol) in dimethylsulfoxide (10 mL) was added 5-bromopyridin-3-ol (71 mg, 0.410 mmol) and potassium carbonate (130 mg, 0.945 mmol). The reaction mixture was stirred at 100° C. for 48 h. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (50 mL). The organics were washed with water (20 mL) and brine (20 mL), dried over sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography eluting with dichloromethane / methanol=15 / 1 to give 4-(4-((5-bromopyridin-3-yl)oxy)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (35 mg, 24%) as a white solid. LCMS (ESI) m / z: 455.0 [M+H] + .

[0292] Step 2: Synthesis of 4-(4-((5-phenylpyridin-3-yl)oxy)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0293] Potassium carbonate (170 mg, 0.522 mmol) was added to a solution of 4-(4-((5-bromopyridin-3-yl)oxy)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (35 mg, 0.077 mmol), phenylboronic acid (19 mg, 0.154 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (5.8 mg, 0.008 mmol) in dioxane (2 mL) and water (0.5 mL) at room temperature. The resulting mixture was stirred at 100° C. under nitrogen atmosphere for 3 hours and cooled. It was then diluted with ethyl acetate (50 mL) and the organic phase was washed with water (20 mL×2) and brine (30 mL), dried over sodium sulfate, filtered and concentrated. The residue was subjected to preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 4-(4-((5-phenylpyridin-3-yl)oxy)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (15.6 mg, 44%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.04 (s, 1H), 8.77 (d, J = 2.0Hz, 1H), 8.50 (d, J = 2.5Hz, 1H), 8.24-8.20 (m, 2H), 7.99-7.97 (m, 1H), 7.77 LCMS (ESI) m / z: 453.1 [M+H] + . Synthesis of 2-morpholino-N-(oxetan-3-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-amine (compound 76): [ka]

[0294] To a solution of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (120 mg, 0.38 mmol) in dioxane (10 mL) was added oxetan-3-amine (55 mg, 0.76 mmol), Pd2(dba)3 (52 mg, 0.057 mmol), Xantphos (66 mg, 0.11 mmol) and Cs2CO3 (371 mg, 1.14 mmol). The resulting mixture was stirred at 110 °C for 16 h and concentrated. The resulting crude product was purified by preparative HPLC (0.05% FA / HO:CHCN=5%-95%) to give 2-morpholino-N-(oxetan-3-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-amine (14.3 mg, 11%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.47 (s, 1H), 7.67 (d, J = 8.0Hz, 2H), 7.40 (t, J = 7.6Hz, 2H), 7.11 (t, J = 7.6Hz, 1H), 4.44 (t, J = 9.6Hz, 1H), 4.21 (t, J = 8.4Hz, 3H), 4.08 (dd, J = 10.4, 5.6Hz, 1H), 3.73 (t, J = 4.4Hz, 4H), 3.57-3.45 (m, 6H), 2.95-2.91 (m, 2H); LCMS (ESI) m / z: 354.1 [M+H]+. Following the protocol described above, the following compounds were synthesized: [Table 12]

[0295] Synthesis of 4,4'-(7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine-2,4-diyl)dimorpholine (compound 78): [ka]

[0296] To a solution of morpholine (45 mg, 0.52 mmol) in THF (10 mL) was added NaH (38 mg, 0.95 mmol) at 0° C. The suspension was stirred at room temperature for 15 min, followed by the addition of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (150 mg, 0.47 mmol) to the mixture. The mixture was then stirred at 80° C. for 16 h, quenched with water (10 mL), extracted with ethyl acetate (30×3 mL), washed with brine (30 mL), dried over sodium sulfate, filtered, and concentrated. The resulting crude product was purified by preparative HPLC (0.05% NH4HCO3 / H2O:CH3CN=5%-95%) to give 4,4'-(7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine-2,4-diyl)dimorpholine (18.6 mg, 11%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 7.72 (d, J = 8.0Hz, 2H), 7.36 (t, J = 8.0Hz, 2H), 7.01 (t, J = 7.6Hz, 1H), 3.98 (t, J = 8.4Hz, 1H), 3.87-3.74 (m, 12H), 3.64-3.62 (m, 4H), 3.16 (t, J = 8.4Hz, 2H); LCMS (ESI) m / z: 368.1 [M+H]+. Synthesis of 8-(2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)octahydropyrazino[2,1-c][1,4]oxazine (compound 79): [ka]

[0297] A mixture of octahydropyrazino[2,1-c][1,4]oxazine hydrochloride (60 mg, 0.337 mmol), 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (107 mg, 0.337 mmol) and cesium carbonate (328 mg, 1.011 mmol) in N,N-dimethylformamide (5 mL) was stirred at 85° C. for 4 h. Then water (10 mL) was added and the mixture was extracted with ethyl acetate (20 mL×3). The organic layer was dried and concentrated. The resulting crude product was purified by SGC (dichloromethane:methanol 50:1 to 10:1) to give 8-(2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)octahydropyrazino[2,1-c][1,4]oxazine (23.2 mg, 16%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.97 (d, J = 2.8Hz, 1H), 8.15-8.10 (m, 2H), 7.34 (dd, J = 7.6, 4.4Hz, 1H), 4.26 (d, J = 14Hz, 1H), 4.09 (d, J = 12.4Hz, 1H), 3.94 (t, J = 8.4Hz, 2H), 3.75-3.71(m, 2H), 3.64-3.52 (m, 9H), 3.31 (s, 1H), 3.17- 3.13 (m, 3H), 2.98-2.97 (m, 1H), 2.76-2.73 (m, 1H), 2.65-2.62 (m, 1H), 2.20-2.11(m, 3H); LC-MS: m / z=424(M+H) + . Following the protocol described above, the following compounds were synthesized: [Table 13]

[0298] Synthesis of (E)-4-(4-(2-(3-methylbenzylidene)hydrazinyl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 82): [ka]

[0299] Step 1: Synthesis of 4-(4-hydrazineyl-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0300] To a suspension of 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 0.157 mmol) in 1,4-dioxane (1.5 mL) was added hydrazine monohydrate (98.5 mg, 1.57 mmol) and the suspension was stirred under reflux for 5 h. After cooling to room temperature, the reaction mixture was treated with water (40 mL) and extracted with ethyl acetate (5 mL×3). The combined organic layers were concentrated to give 4-(4-hydrazineyl-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (70 mg, 99%). LCMS (ESI) m / z: 314.3 [M+H] + .

[0301] Step 2: Synthesis of (E)-4-(4-(2-(3-methylbenzylidene)hydrazinyl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0302] To a solution of 4-(4-hydrazineyl-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 0.16 mmol), 3-methylbenzaldehyde (38 mg, 0.32 mmol) in ethanol (10 mL) was added acetic acid (0.05 mL). The mixture was refluxed at 80° C. for 16 hours, then diluted with water (80 mL) and extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (150 mL), dried over sodium sulfate, filtered, and concentrated. The residue was subjected to preparative HPLC to give (E)-4-(4-(2-(3-methylbenzylidene)hydrazinyl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (12.5 mg, 19% for two steps) as a yellow solid. 1H NMR (400 MHz, DMSO-d6) δ 10.77 (s, 1H), 9.04 (d, J = 2.5Hz, 1H), 8.18 (t, J = 6.8Hz, 2H), 7.99 (s, 1H), 7.58 - 7.28 (m, 4H), 7.16 (d, LCMS (ESI) m / z: 416.2 [M+H] + . Synthesis of 4-(7-(pyridin-3-yl)-4-(3-(m-tolyl)-1H-pyrazol-1-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 83): [ka]

[0303] Step 1: Synthesis of (E)-3-(dimethylamino)-1-(m-tolyl)prop-2-en-1-one.

[0304] A solution of 1-(m-tolyl)ethan-1-one (500 mg, 3.72 mmol) in N,N-dimethylformamide dimethyl acetal (5 mL) was stirred at 110° C. for 17 h. The mixture was concentrated to give (E)-3-(dimethylamino)-1-(m-tolyl)prop-2-en-1-one as a yellow oil. LCMS (ESI) m / z: 190.2 [M+H] + This product was used in the next step without further purification.

[0305] Step 2: Synthesis of 3-(m-tolyl)-1H-pyrazole.

[0306] A mixture of (E)-3-(dimethylamino)-1-(m-tolyl)prop-2-en-1-one (567.78 mg, 3 mmol) and hydrazine hydrate (563.18 mg, 9 mmol) in ethanol (6 mL) was stirred at reflux for 2 h. The reaction mixture was concentrated to give 3-(m-tolyl)-1H-pyrazole as a yellow oil (450 mg, 94% for two steps). LCMS (ESI) m / z: 159.1 [M+H] + .

[0307] Step 3: Synthesis of 4-(7-(pyridin-3-yl)-4-(3-(m-tolyl)-1H-pyrazol-1-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0308] A mixture of 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 0.16 mmol), 3-(m-tolyl)-1H-pyrazole (38 mg, 0.32 mmol) and cesium carbonate (156 mmol, 0.48 mg) in N,N-dimethylacetamide (4 mL) was stirred at 120° C. for 16 h. The mixture was filtered and the filtrate was purified by preparative HPLC to give 4-(7-(pyridin-3-yl)-4-(3-(m-tolyl)-1H-pyrazol-1-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (12.4 mg, 18%) as a white solid.

[0315] 1H NMR (400 MHz, DMSO-d6) δ 9.10 (d, J = 2.3Hz, 1H), 8.73 (d, J = 2.7Hz, 1H), 8.47 - 8.04 (m, 2H), 7.77 (d, J = 9.5Hz, 2H), 7.44 (dd, J = 8.4, 4.6Hz, 1H), 7.36 (t, J = 7.6Hz, 1H), 7.21 (d, J = 8.2Hz, 1H), 7.06 (d, J = 2.7Hz, 1H), 4.19 (t, J = 8.3Hz, 2H), 3.78-3.70 (m, 8H), 3.57 (t, J = 8.5Hz, 2H), 2.39 (s, J = 1H).; LCMS (ESI) m / z: 440.1 [M+H] + . Synthesis of 4-(4-(3-phenyl-1H-pyrazol-1-yl)-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 84): [ka]

[0309] Step 1: Synthesis of 4-(4-chloro-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0310] To a solution of pyridin-4-amine (517 mg, 5.5 mmol) in tetrahydrofuran (50 mL) was added sodium hydride (400 mg, 10.0 mmol) slowly at 0° C. The mixture was stirred at 80° C. for 2 h and a solution of 4-(4,6-dichloro-5-(2-chloroethyl)pyrimidin-2-yl)morpholine (1.5 g, 5.0 mmol) in THF (3 mL) was added. The resulting mixture was stirred at 80° C. for another 16 h. It was poured into crushed ice and extracted with ethyl acetate (150×2 mL). The combined organic layers were concentrated and the resulting crude product was purified by silica gel column chromatography [(20% dichloromethane in methanol) and further washed with methanol (10 mL)] to give 4-(4-chloro-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (280 mg, 18%) as a grey solid. LCMS (ESI) m / z: 318.1 / 320.1 [M+H] + .

[0311] Step 2: Synthesis of 4-(4-(3-phenyl-1H-pyrazol-1-yl)-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0312] A mixture of 4-(4-chloro-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (63 mg, 0.2 mmol), 3-phenyl-1H-pyrazole (43 mg, 0.3 mmol) and cesium carbonate (130 mg, 0.4 mmol) in N,N-dimethylformamide (5 mL) was stirred at 110° C. for 4 hours. It was then poured into water and the resulting precipitate was collected by filtration, washed with methanol (10 mL) and dried under vacuum to give 4-(4-(3-phenyl-1H-pyrazol-1-yl)-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (40 mg, 47%) as a yellow solid. 1H NMR (400 MHz, DMSO-d6) δ 8.75 (d, J = 2.7Hz, 1H), 8.47 (d, J = 5.4Hz, 2H), 7.97 (d, J = 7.3Hz, 2H), 7.83 (d, J = 6.1Hz, 2H), 7.48 (t, J = 7.5Hz, 2H), 7.39 (s, 1H), 7.09 (d, J = 2.7Hz, 1H), 4.13 (t, J = 8.3Hz, 2H), 3.78 (d, J = 4.8Hz, 4H), 3.73 (d, J = 4.8Hz, 4H), 3.55 (t, J = 8.2Hz, 2H); LCMS (ESI) m / z: 425.9 [M] + . Synthesis of 4-(4-((3-methoxyphenyl)ethynyl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 85): [ka]

[0313] To a suspension of 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 0.157 mmol), bis(triphenylphosphine)palladium(II) chloride (11 mg, 0.0157 mmol) and cuprous iodide (6 mg, 0.0315 mmol) in triethylamine (0.5 mL) was added 1-ethynyl-3-methoxybenzene (51 mg, 0.393 mmol) using a syringe under argon atmosphere and the resulting mixture was heated at 70° C. for 16 h. The mixture was filtered and the filtrate was washed with a saturated solution of ammonium chloride (25 mL×2) and water (25 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was subjected to preparative HPLC to give 4-(4-((3-methoxyphenyl)ethynyl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (11.1 mg, 17%) as a yellow solid. 1H NMR (400 MHz, CD3OD) δ 9.22 (s, 1H), 8.26 (d, J = 8.9Hz, 2H), 7.50-7.48 (m, 1H), 7.35 (t, J = 8.0Hz, 1H), 7.19 - 7.14 (m, 2H), 7.05-7.02 (m, J = 8.0Hz, 1H), 4.19 (t, J = 8.0Hz, 2H), 3.87 (s, 3H), 3.78 (s, 8H), 3.25 (t, J = 8.0Hz, 2H); LCMS (ESI) m / z: 414.0 [M+H] + . Synthesis of 4-(7-phenyl-4-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 86): [ka]

[0314] Step 1: Synthesis of 4-(4-(3,6-dihydro-2H-pyran-4-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0315] A mixture of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (100 mg, 0.32 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (199 mg, 0.95 mmol), CsCO (309 mg, 0.95 mmol), Pd(dba) (29 mg, 0.03 mmol) and P(Cy) (17 mg, 0.06 mmol) in DMSO (10 mL) / HO (2 mL) was stirred at 140° C. for 16 h under nitrogen atmosphere. The reaction was then quenched with water (10 mL) and the mixture was extracted with EtOAc (20×3 mL). The organic layers were combined, washed with brine (30 mL), dried over Na2SO4, filtered and concentrated. The residue was subjected to preparative TLC (PE / EA=4:1) to give 4-(4-(3,6-dihydro-2H-pyran-4-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (70 mg, 60%) as a yellow solid. LCMS (ESI) m / z: 365.2 [M+H] + .

[0316] Step 2: Synthesis of 4-(7-phenyl-4-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0317] A suspension of 4-(4-(3,6-dihydro-2H-pyran-4-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 0.14 mmol) and 10% Pd / C (5 mg) in MeOH (10 mL) was stirred at room temperature under hydrogen atmosphere for 1 h. The mixture was then filtered, concentrated, and subjected to preparative HPLC (0.05% FA / HO:CHCN=5%-95%) to give 4-(7-phenyl-4-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (26.6 mg, 52%) as a yellow solid. 1H NMR (400 MHz, DMSO-d6) δ 7.80 (d, J = 8.0Hz, 2H), 7.37 (t, J = 8Hz, 2H), 7.01 (t, J = 7.2Hz, 1H), 4.04 (t, J = 8.4Hz, 2H), 3.93 (dd, J = 11.6, 3.2Hz, 2H), 3.66 (s, 8H), 3.43 (t, J = 10.8Hz, 2H), 3.02 (t, J = 8.4Hz, 2H), 2.75 (t, J = 11.6Hz, 1H), 1.87-1.83 (m, 2H), 1.57 (d, J = 11.2Hz, 2H); LCMS (ESI) m / z: 367.2 [M+H]+. Synthesis of 4-(7-phenyl-4-(tetrahydrofuran-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 87): [ka]

[0318] Step 1: Synthesis of 4-(4-(furan-3-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0319] A mixture of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (85 mg, 0.27 mmol), furan-3-ylboronic acid (90 mg, 0.81 mmol), Cs2CO3 (263 mg, 0.81 mmol) and Pd(dppf)Cl2 (19 mg, 0.027 mmol) in DMSO (8 mL) / HO (2 mL) was stirred at 130° C. for 16 h under nitrogen atmosphere. The mixture was then diluted with EtOAc (45 mL), washed with water (15 mL), brine (30 mL), dried over Na2SO4, filtered and concentrated. The residue was subjected to SGC (PE / EA=4:1) to give 4-(4-(furan-3-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 54%) as a yellow solid; LCMS (ESI) m / z: 349.1 [M+H] + .

[0320] Step 2: Synthesis of 4-(7-phenyl-4-(tetrahydrofuran-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0321] A suspension of 4-(4-(furan-3-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 0.14 mmol) and 10% Pd / C (5 mg) in MeOH (15 mL) was stirred at room temperature under hydrogen atmosphere for 2 h. The mixture was then filtered and the filtrate was concentrated. The residue was subjected to preparative HPLC (0.05% FA / H2O:CH3CN=5%-95%) to give 4-(7-phenyl-4-(tetrahydrofuran-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (13.3 mg, 26%) as a yellow solid. 1H NMR (400 MHz, DMSO-d6) δ 7.80 (d, J = 7.6Hz, 2H), 7.37 (t, J = 8.0Hz, 2H), 7.02 (t, J = 7.4Hz, 1H), 4.07-3.99 (m, 3H), 3.92-3.86 (m, LCMS (ESI) m / z: 353.1 [M+H]+.

[0322] Synthesis of 4-(4-(furan-3-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 88) and 4-(7-(pyridin-3-yl)-4-(tetrahydrofuran-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 89): [ka]

[0323] Step 1: Synthesis of 4-(4-(furan-3-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0324] To a stirred mixture of 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (200 mg, 0.62 mmol), furan-3-ylboronic acid (211 mg, 1.88 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (69 mg, 0.094 mmol) in acetonitrile (6 mL) and water (1.5 mL) was added cesium carbonate (615 mg, 1.88 mmol) at 20° C. The resulting mixture was stirred at 80° C. under nitrogen for 18 h and cooled. The reaction mixture was then quenched with water (50 mL) and extracted with dichloromethane (50 mL×2). The combined organic fractions were washed with brine (50 mL), dried over sodium sulfate, filtered, and concentrated. The residue was subjected to preparative HPLC to give the title compound (28.1 mg, 13%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 9.13 (s, 1H), 8.29 (d, J = 3.5Hz, 1H), 8.21 (d, J = 8.7Hz, 1H), 7.95 (s, 1H), 7.51 (t, J = 1.7Hz, 1H), 7.34 (dd, J = 8.5, 4.7Hz, 1H), 6.95 (s, 1H), 4.14 (t, J = 8Hz, 2H), 3.84-3.80 (m, 98), 3.24 (t, J = 8Hz, 2H); LCMS (ESI) m / z: 350.1 [M+H] +

[0325] Step 2: Preparation of 4-(7-(pyridin-3-yl)-4-(tetrahydrofuran-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0326] To a solution of 4-(4-(furan-3-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (74 mg, 0.21 mmol) in methanol (25 mL) and acetate (25 mL) was added palladium on activated carbon 10% Pd (74 mg). The resulting suspension was stirred at 50° C. for 5 h under a hydrogen atmosphere. The reaction mixture was filtered and the filtrate was concentrated. The residue was subjected to preparative HPLC (BOSTON pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile / 0.1% ammonium bicarbonate) to give 4-(7-(pyridin-3-yl)-4-(tetrahydrofuran-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (16.1 mg, 21%) as a white solid. 1 H NMR (500 MHz, Chloroform-d) δ 9.08 (d, J = 2.8Hz, 1H), 8.29 (dd, J = 4.0, 0.4Hz, 1H), 8.15 (dt, J = 9.2, 0.8Hz, 1H), 7.30 (dd, J = 8.5, 4.7Hz, 1H), 4.11 (t, J = 8.0Hz, 1H), 4.08 - 4.00 (m, 3H), 3.96 - 3.88 (m, 2H), 3.80 - 3.75 (m, 8H), 3.36 (pent, J = 6.4Hz, 1H), 3.14 - 3.02 (m, 2H), 2.34 - 2.26 (m, 1H), 2.22 - 2.14 (m, 1H). LCMS (ESI) m / z: 354.2 [M+H] + . Synthesis of 4-(7-phenyl-4-(pyridin-2-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 90): [ka]

[0327] A solution of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (250 mg, 0.79 mmol), pyridin-2-ylboronic acid (486 mg, 3.95 mmol), 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex (131 mg, 0.16 mmol) and cesium carbonate (772 mg, 2.37 mmol) in water (5.0 mL) and DMSO (20 mL) was stirred under argon atmosphere at 130° C. for 8 h. The mixture was diluted with ethyl acetate (150 mL), washed with water (150 mL) and the organic layer was concentrated. The resulting crude product was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 4-(7-phenyl-4-(pyridin-2-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine as a yellow solid (44.3 mg, 15%). 1 H NMR (400 MHz, Chloroform-d) δ 8.67 (d, J = 4.9, 1.7Hz, 1H), 8.37 (d, J = 8.0Hz, 1H), 7.90 - 7.72 (m, 3H), 7.42 (t, J = 8.0Hz, 2H), 7.30 - 7.27 (m, 1H), 7.06 (t, J = 7.4Hz, 1H), 4.12 (t, J = 8.3Hz, 2H), 3.94 - 3.75 (m, 8H), 3.60 (t, J = 8.3Hz, 2H). LCMS (ESI) m / z: 360.2 [M+H] + .

[0328] Following the above protocol, the following compounds were synthesized: [Table 14-1] [Table 14-2]

[0329] Synthesis of 4-(6-methyl-7-(pyridin-4-yl)-4-(tetrahydrofuran-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 98): [ka]

[0330] Step 1: Synthesis of methyl 5-methyl-2-oxotetrahydrofuran-3-carboxylate.

[0331] A solution of methyldihydrofuran-2(3H)-one (25 g, 250 mmol) in tetrahydrofuran (100 mL) was added dropwise to lithium bis(trimethylsilyl)amide (1.6 M in tetrahydrofuran, 330 mL, 528 mmol) at -78°C. After stirring at -78°C for 10 min, dimethyl carbonate (23.6 g, 263 mmol) was added to the mixture at -78°C, and the reaction mixture was warmed and stirred at room temperature for 16 h. It was then poured into a mixture of concentrated hydrochloric acid (80 mL) and ice (800 mL), followed by extraction with ethyl acetate (800 mL x 2). The organic layer was washed with brine, dried over sodium sulfate, filtered and concentrated to give the title compound (40 g). This product was used in the next step without further purification. LCMS (ESI) m / z: 159.1 [M+H] + .

[0332] Step 2: Synthesis of 5-(2-hydroxypropyl)-2-morpholinopyrimidine-4,6-diol.

[0333] Methyl 5-methyl-2-oxotetrahydrofuran-3-carboxylate (40 g, 250 mmol) was added to a solution of morpholine-4-carboximidamide hydrochloride (31 g, 192 mmol) and sodium methanolate (104 g, 576 mmol) in methanol (150 mL) at room temperature. The reaction mixture was then refluxed for 16 h and cooled. Water (200 mL) was added to the mixture and stirred for 0.5 h, followed by acetic acid (30 mL) and the mixture was further stirred at room temperature for 2 h. The precipitated solid was filtered and dried to give the title compound (41 g, 83%) as a white solid. LCMS (ESI) m / z: 256.2 [M+H] + .

[0334] Step 3: Synthesis of 4-(4,6-dichloro-5-(2-chloropropyl)pyrimidin-2-yl)morpholine.

[0335] To a solution of 5-(2-hydroxypropyl)-2-morpholinopyrimidine-4,6-diol (41 g, 81 mmol) and N-ethyl-N-isopropylpropan-2-amine (44 mL) in toluene (400 mL) was added phosphorus oxychloride (64 mL) at room temperature. The resulting mixture was stirred at 110° C. for 16 hours and concentrated. The residue was then dissolved in ethyl acetate (1600 mL), washed with water (300 mL×2), brine (300 mL), and dried over sodium sulfate. After concentration, the resulting residue was purified on silica gel column chromatography (petroleum ether / ethyl acetate=10 / 1) to give the title compound (38 g, 76%) as an off-white solid. LCMS (ESI) m / z: 312.0 [M+H] + .

[0336] Step 4: Synthesis of 4-(4-chloro-6-methyl-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0337] A mixture of pyridin-4-amine (151 mg, 1.60 mmol) and sodium hydride (161 mg, 4.025 mmol) in tetrahydrofuran (40 mL) was refluxed for 1 h. After the mixture was cooled to room temperature, 4-(4,6-dichloro-5-(2-chloropropyl)pyrimidin-2-yl)morpholine (500 mg, 1.61 mmol) was added. The resulting mixture was refluxed for 16 h, then poured onto ice water (80 mL) and extracted with ethyl acetate (120 mL x 2). The organic layer was washed with brine (50 mL) and dried over sodium sulfate. This was filtered and concentrated, and the residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate=1 / 1 to 0 / 100) to obtain 4-(4-chloro-6-methyl-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (200 mg, 37%) as a brown solid. LCMS(ESI)m / z:332.2[M+H] + . Step 5: Synthesis of 4-(4-(furan-3-yl)-6-methyl-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine

[0338] To a stirred mixture of 4-(4-chloro-6-methyl-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (200 mg, 0.603 mmol), furan-3-ylboronic acid (135 mg, 1.207 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II):CHCl (49 mg, 0.06 mmol) in acetonitrile (8 mL) and water (2 mL) was added cesium carbonate (393 mg, 1.206 mmol) at room temperature. The resulting reaction mixture was stirred at 80° C. under nitrogen for 4 h and cooled. The reaction was quenched with water (50 mL) and the mixture was extracted with dichloromethane (100 mL×2). The combined extracts were washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. The resulting residue was subjected to silica gel column chromatography eluting with petroleum ether / ethyl acetate=1 / 1 to give 4-(4-(furan-3-yl)-6-methyl-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (200 mg, 91%) as a yellow solid. LCMS(ESI)m / z:364.0[M+H] + .

[0339] Step 6: Synthesis of 4-(6-methyl-7-(pyridin-4-yl)-4-(tetrahydrofuran-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0340] A suspension of 4-(4-(furan-3-yl)-6-methyl-7-(pyridin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (200 mg, 0.55 mmol) and palladium on activated carbon (10%, 100 mg) in methanol (20 mL) and ethyl acetate (10 mL) was stirred under a hydrogen atmosphere for 16 h at 30° C. The mixture was filtered through Celite and concentrated. The resulting residue was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 4-(6-methyl-7-(pyridin-4-yl)-4-(tetrahydrofuran-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (17.1 mg, 8%) as a white solid. 1 H NMR (500 MHz, MeOD) δ 8.42 (d, J = 6.0Hz, 2 H), 7.83 (d, J = 6.0Hz, 2 H), 4.80 (bs, 1H), 4.01-3.97 (m, 1H), 3.89-3.87 (m, 1H), 3.82-3.62 LCMS (ESI) m / z: 368.1 [M+H] + . Synthesis of 4-(7-phenyl-4-(pyridazin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 99): [ka]

[0341] A solution of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (60 mg, 0.19 mmol), 4-(tributylstannyl)pyridazine (70 mg, 0.19 mmol), LiCl (8 mg, 0.19 mmol) and Pd(PPh3)4 (22 mg, 0.019 mmol) in dioxane (5 mL) was stirred for 16 hours at 100 ° C. under a nitrogen atmosphere. This was concentrated, and the residue was subjected to preparative HPLC (0.05% NH4HCO3 / H2O: CH3CN = 5% to 95%) to obtain 4-(7-phenyl-4-(pyridazin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (1.8 mg, 3%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 9.74 (d, J = 0.8Hz, 1H), 9.33 (dd, J = 5.2, 1.2Hz, 1H), 8.00 (dd, J = 5.2, 2.4Hz, 1H), 7.81 (d, J = 8.0Hz, 2H), 7.44 (t, J = 8.0Hz, 2H), 7.14 (t, J = 7.6Hz, 1H), 4.20 (t, J = 8.0Hz, 2H), 3.90-3.82 (m, 8H), 3.42 (t, J = 8.0Hz, 2H); LCMS (ESI) m / z: 361.2 [M+H]+. [Table 15-1] [Table 15-2]

[0342] Synthesis of tert-butyl 4-(2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-1-carboxylate (compound 107), 4-(4-(piperidin-4-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 108) and 4-(4-(1-methylpiperidin-4-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 109): [ka]

[0343] Step 1: Synthesis of tert-butyl 4-(2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-5,6-dihydropyridine-1(2H)-carboxylate.

[0344] A mixture of 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 0.157 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate (97 mg, 0.314 mmol), tris(dibenzylideneacetone)dipalladium (15 mg, 0.016 mmol), tricyclohexylphosphine (9 mg, 0.032 mmol) and cesium carbonate (103 mg, 0.316 mmol) in acetonitrile (8 mL) and water (2 mL) was refluxed under nitrogen for 16 h. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (80 mL), washed with water (30 mL) and brine (20 mL), dried over sodium sulfate, filtered and concentrated. The resulting crude product was purified by silica gel column chromatography eluting with petroleum ether / ethyl acetate=1 / 1 then 0 / 100 to give tert-butyl 4-(2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-5,6-dihydropyridine-1(2H)-carboxylate (50 mg, 68%) as a brown solid. LCMS (ESI) m / z: 465.3 [M+H] + .

[0345] Step 2: Synthesis of tert-butyl 4-(2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-1-carboxylate.

[0346] A suspension of tert-butyl 4-(2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-5,6-dihydropyridine-1(2H)-carboxylate (50 mg, 0.108 mmol) and palladium on activated carbon (10%, 30 mg) in methanol (5 mL) and ethyl acetate (5 mL) was stirred at room temperature under a hydrogen atmosphere for 5 h. The resulting mixture was filtered through Celite and concentrated. The resulting crude product was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give tert-butyl 4-(2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-1-carboxylate (4.9 mg, 10%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.03 (d, J = 2.4Hz, 1H), 8.22-8.19 (m, 2H), 7.40-7.37 (m, 1H), 4.09-4.03 (m, 4H), 3.65 (s, 8H), 3.05 (t, J = 8.4Hz, 2H), 2.89-2.67 (m, 3H), 1.67-1.61 (m, 4H), 1.41 (s, 9H). LCMS (ESI) m / z: 467.2 [M+H] + .

[0347] Step 3: Synthesis of 4-(4-(piperidin-4-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0348] Trifluoroacetic acid (1 mL) was added to a solution of tert-butyl 4-(2-morpholino-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-1-carboxylate (50 mg, 0.107 mmol) in dichloromethane (2 mL) at room temperature. The mixture was stirred at room temperature for 2 hours, then concentrated and the resulting residue was subjected to preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 minutes at 2 ml / min, and the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 4-(4-(piperidin-4-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (12.3 mg, 31%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.04 (d, J = 2.4Hz, 1H), 8.22-8.19 (m, 2H), 7.40-7.37 (m, 1H), 4.01 (t, J = 8.4Hz, 2H), 3.67 (s, 8H), 3.14-3.11 (m, 2H), 3.04 (t, J = 8.4Hz, 2H), 2.74-2.66 (m, 3H), 1.83-1.64 (m, 4H). LCMS (ESI) m / z: 367.3 [M+H] + .

[0349] Step 4: Synthesis of 4-(4-(1-methylpiperidin-4-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0350] Acetic acid (16 mg, 0.266 mmol) was added to a mixture of 4-(4-(piperidin-4-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 0.136 mmol) and formaldehyde solution (37 wt% in water) (1 mL) in methanol (10 mL) at room temperature. After stirring at room temperature for 2 h, sodium cyanoborohydride (17 mg, 0.271 mmol) was added to the mixture and further stirred for another 2 h. This was then concentrated and the resulting residue was subjected to preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give 4-(4-(1-methylpiperidin-4-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (34.7 mg, 67%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.04 (s, 1H), 8.21-8.20 (m, 2H), 7.40-7.37 (m, 1H), 4.06 (t, J = 8.2Hz, 2H), 3.66 (s, 9H), 3.03 (t, J = LCMS (ESI) m / z: 381.3 [M+H] + .

[0351] Synthesis of tert-butyl 4-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-1-carboxylate (compound 110), 4-(7-phenyl-4-(piperidin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 111) and 4-(4-(1-methylpiperidin-4-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 112): [ka]

[0352] Step 1: Synthesis of tert-butyl 4-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,6-dihydropyridine-1(2H)-carboxylate.

[0353] A mixture of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (200 mg, 0.63 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (391 mg, 1.26 mmol), CsCO (619 mg, 1.90 mmol), Pd(dba) (58 mg, 0.063 mmol) and P(Cy) (35 mg, 0.126 mmol) in CHCN (20 mL) / HO (5 mL) was stirred at 100° C. under nitrogen atmosphere for 4 h. The mixture was concentrated and the residue was extracted with EtOAc (20×3 mL) / HO (10 mL). The organic layer was washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. The residue was purified by SGC (PE / EA=4:1) to give tert-butyl 4-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,6-dihydropyridine-1(2H)-carboxylate (160 mg, 55%) as a yellow solid. LCMS (ESI) m / z: 464.3 [M+H]+ .

[0354] Step 2: Synthesis of tert-butyl 4-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-1-carboxylate.

[0355] To a solution of tert-butyl 4-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,6-dihydropyridine-1(2H)-carboxylate (160 mg, 0.35 mmol) in MeOH (20 mL), 10% Pd / C (16 mg) was added, and the resulting mixture was stirred at room temperature under hydrogen atmosphere for 1 h. The mixture was filtered and concentrated, and the resulting crude product was purified by preparative HPLC (0.05% NH4HCO3 / H2O:CH3CN=5%-95%) to give tert-butyl 4-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-1-carboxylate (130 mg, 81%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 7.78 (d, J = 8.0Hz, 2H), 7.39 (t, J = 8.0Hz, 2H), 7.05 (t, J = 7.2Hz, 1H), 4.22 (bs, 2H), 4.06 (t, J = 8.4Hz, 2H), 3.79 (s, 8H), 3.06-3.02 (m, 2H), 2.86-2.79 (m, 2H), 2.61-2.60 (m, 1H), 1.89-1.82 (m, 2H), 1.73-1.69 (m, 2H), 1.51 (s, 9H); LCMS (ESI) m / z: 466.2 [M+H]+.

[0356] Step 3: Synthesis of 4-(7-phenyl-4-(piperidin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0357] To a solution of tert-butyl 4-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-1-carboxylate (100 mg, 0.2 mmol) in dichloromethane (2 mL) was added TFA (0.5 mL) at 0° C. The mixture was then stirred at room temperature for 2 h and concentrated. The residue was purified by preparative HPLC (0.05% NH4HCO3 / H2O:CH3CN=5%-95%) to give 4-(7-phenyl-4-(piperidin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (55 mg, 70%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 7.79 (d, J = 8.0Hz, 2H), 7.38 (t, J = 8.0Hz, 2H), 7.05 (t, J = 7.2Hz, 1H), 4.06 (t, J = 8.0Hz, 2H), 3.80 (s, 8H), 3.27 (d. (ESI) m / z: 366.1 [M+H]+.

[0358] Step 4: Synthesis of 4-(4-(1-methylpiperidin-4-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0359] To a solution of 4-(7-phenyl-4-(piperidin-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (40 mg, 0.11 mmol) in methanol (5 mL) was added formaldehyde (4 mg, 0.12 mmol). The mixture was stirred at room temperature for 2 h, followed by the addition of sodium cyanoborohydride (35 mg, 0.55 mmol) to the mixture. The mixture was further stirred at room temperature for 12 h and concentrated. The resulting crude product was purified by preparative HPLC (0.05% NH4HCO3 / H2O:CH3CN=5%-95%) to give 4-(4-(1-methylpiperidin-4-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (35.9 mg, 85%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 7.78 (d, J = 8.0Hz, 2H), 7.38 (t, J = 8.0Hz, 2H), 7.04 (t, J = 7.2Hz, 1H), 4.05 (t, J = 8.4Hz, 2H), 3.79 (s, 8H), 3.06-2.99 (m, 4H), 2.43 (bs, 1H), 2.34 (s, 3H), 2.09-2.00 (m, 4H), 1.74-1.63 (m, 2H); LCMS (ESI) m / z: 380.3 [M+H]+. Synthesis of tert-butyl 3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)azetidine-1-carboxylate (compound 113): [ka]

[0360] A solution of (1-(tert-butoxycarbonyl)azetidin-3-yl)zinc(II) iodide (0.5 M in N,N-dimethylacetamide) (1.264 mL, 0.632 mmol) was added to a solution of 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (50 mg, 0.158 mmol) and bis(tri-tert-butylphosphine)palladium(0) (16 mg, 0.031 mmol) in N,N-dimethylacetamide (2 mL) at room temperature. The resulting mixture was stirred at 80° C. for 16 hours and then quenched with saturated ammonium chloride solution (10 mL). The mixture was then extracted with ethyl acetate (20 mL×3) and the combined organic layers were washed with water (20 mL×2), brine (20 mL), dried over sodium sulfate, filtered and concentrated. The crude product was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The elution system used was a 5% to 95% gradient over 1.5 min at 2 ml / min, the solvent was acetonitrile / 0.01% aqueous ammonium bicarbonate) to give tert-butyl 3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)azetidine-1-carboxylate (4.7 mg, 7%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.82 (d, J = 8.0Hz, 2H), 7.36 (t, J = 7.6Hz, 2H), 7.03 (t, J = 7.2Hz, 1H), 4.21-4.17 (m, 4H), 4.08 (t, J = 8.4Hz, 2H), 3.80-3.73 (m, 9H), 3.00 (t, J = 8.4Hz, 2H), 1.48 (s, 9H). LCMS (ESI) m / z: 438.1 [M+H] + . Following the protocol described above, the following compounds were synthesized: [Table 16]

[0361] Synthesis of cyclopropyl(3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrrolidin-1-yl)methanone (compound 118): [ka]

[0362] Step 1: Synthesis of tert-butyl 3-(((trifluoromethyl)sulfonyl)oxy)-2,5-dihydro-1H-pyrrole-1-carboxylate.

[0363] A solution of tert-butyl 3-oxopyrrolidine-1-carboxylate (1400 mg, 7.567 mmol) and N,N-diisopropylethylamine (2928 mg, 22.701 mmol) in dichloromethane (50 mL) was cooled to -78°C and stirred for 10 min. Trifluoromethanesulfonic anhydride (2560 mg, 9.081 mmol) was then added and the mixture was warmed and stirred at 25°C for 16 h. The reaction was quenched with aqueous ammonium chloride and extracted with dichloromethane (50 mL x 3). The organic layer was dried and concentrated to give tert-butyl 3-(((trifluoromethyl)sulfonyl)oxy)-2,5-dihydro-1H-pyrrole-1-carboxylate (800 mg, 33%) as a yellow oil. LC-MS: m / z=262 (M-56+H) + .

[0364] Step 2: Synthesis of tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate.

[0365] A solution of tert-butyl 3-(((trifluoromethyl)sulfonyl)oxy)-2,5-dihydro-1H-pyrrole-1-carboxylate (2800 mg, 8.832 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (4487 mg, 17.665 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (325 mg, 0.441 mmol) and potassium acetate (2600 mg, 26.532 mmol) in dioxane (80 mL) was stirred at 75° C. for 4 hours. Water was then added and the resulting mixture was extracted with ethyl acetate (50 mL×3). The organic layer was dried and concentrated. The resulting crude product was purified by silica gel column (petroleum ether:ethyl acetate 50:1 to 10:1) to give tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (2050 mg, 78%) as a yellow solid. LC-MS: m / z=240 (M-56+H). + .

[0366] Step 3: Synthesis of tert-butyl 3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate.

[0367] A solution of tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (280 mg, 0.95 mmol), 4-(4-chloro-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (200 mg, 0.63 mmol), bis(diphenylphosphino)ferrocene]dichloropalladium(II) (20 mg, 0.03 mmol) and potassium carbonate (260 mg, 1.89 mmol) in dioxane / water (30 mL) was stirred at 85° C. for 4 hours. Water was then added and the mixture was extracted with ethyl acetate (50 mL×3). The organic layer was dried and concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate 50:1 to 10:1) to give tert-butyl 3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (200 mg, 71%) as a yellow solid. LC-MS: m / z=450 (M+H). + .

[0368] Step 4: Synthesis of tert-butyl 3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrrolidine-1-carboxylate.

[0369] A suspension of tert-butyl 3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (200 mg, 0.445 mmol) and palladium on carbon (100 mg) in methanol (5 mL) was stirred at 25° C. for 16 h. The mixture was filtered and the filtrate was concentrated and dried to give tert-butyl 3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrrolidine-1-carboxylate (180 mg, 90%) as a yellow solid. LC-MS: m / z=452 (M+H). + .

[0370] Step 5: Synthesis of 4-(7-phenyl-4-(pyrrolidin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0371] A solution of tert-butyl 3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrrolidine-1-carboxylate (420 mg, 0.931 mmol) and HCl in dioxane (4 mL) in dichloromethane (6 mL) was stirred at 25° C. for 2 h. The mixture was concentrated to give 4-(7-phenyl-4-(pyrrolidin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (280 mg, 85%) as a yellow solid. LC-MS: m / z=352 (M+H). + .

[0372] Step 6: Synthesis of cyclopropyl(3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrrolidin-1-yl)methanone.

[0373] A solution of 4-(7-phenyl-4-(pyrrolidin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (80 mg, 0.228 mmol) and triethylamine (69 mg, 0.684 mmol) in dichloromethane (5 mL) was stirred at 25° C. for 10 minutes. Cyclopropanecarbonyl chloride (28 mg, 0.274 mmol) was then added and the resulting mixture was stirred at room temperature for 2 hours. It was filtered and the filtrate was concentrated. The resulting crude product was purified by preparative HPLC (column Xbridge 21.2×250 mm C18, 10 um, mobile phase A: water (10 mmol / L ammonium bicarbonate) B: acetonitrile) to give cyclopropyl (3-(2-morpholino-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrrolidin-1-yl)methanone (43.1 mg, 45%) as a yellow solid. 1H NMR (400 MHz, DMSO-d6) δ 7.80 (d, J = 8.0hz, 2H), 7.37 (t, J = 7.2Hz, 2H), 7.01 (t, J = 7.2Hz, 1H), 4.08 -4.05 (m, 2H), 3.93-3.84 (m, LC-MS: m / z=420(M+H) + . Following the protocol described above, the following compounds were synthesized: [Table 17-1] [Table 17-2]

[0374] Synthesis of 4-(4-(1-methylpyrrolidin-3-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 121): [ka]

[0375] A mixture of 4-(7-phenyl-4-(pyrrolidin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (70 mg, 0.112 mmol), formaldehyde (13 mg, 0.398 mmol) and sodium cyanoborohydride (25 mg, 0.398 mmol) in methanol (4 mL) was stirred at 25° C. for 2 h. Then water (10 mL) was added and the mixture was extracted with ethyl acetate (30 mL×3). The organic layer was dried over sodium sulfate, filtered and concentrated. The resulting crude product was purified by silica gel column (dichloromethane:methanol 50:1 to 10:1) to give 4-(4-(1-methylpyrrolidin-3-yl)-7-phenyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (22.4 mg, 28%). 1 H NMR (400 MHz, DMSO-d6) δ 7.80 (d, J = 8.0Hz, 2H), 7.36 (t, J = 7.6Hz, 2H), 7.00 (t, J = 7.2Hz, 1H), 4.03 (t, J = 8.4Hz, 2H), 3.66 (s, 8H), 3.31-3.27(m, 2H), 3.01-2.92 (m, 3H), 2.79-2.78 (m, 1H), 2.48-2.44 (m, 1H), 2.31 (s, 3H), 2.09-2.04 (m, 2H); LC-MS: m / z=366.3(M+H) +. Synthesis of 4-(4-(2-(3-methoxyphenyl)cyclopropyl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 122): [ka]

[0376] Step 1: Synthesis of (E)-4-(4-(3-methoxystyryl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0377] A mixture of 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (317 mg, 1.0 mmol), 1-methoxy-3-vinylbenzene (134 mg, 1.0 mmol), palladium(II) acetate (23 mg, 0.1 mmol), tri(o-tolyl)phosphine (60 mg, 0.2 mmol) and triethylamine (202 mg, 2.0 mmol) in N,N-dimethylformamide (10 mL) was stirred at 120° C. for 16 hours under nitrogen atmosphere. The mixture was poured into water and extracted with ethyl acetate (100 mL×2). The combined organic phase was concentrated and the residue was subjected to silica gel column chromatography to give (E)-4-(4-(3-methoxystyryl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (220 mg, 0.53 mmol) as a yellow solid. LCMS (ESI) m / z: 416.1 [M+H] + .

[0378] Step 2: Synthesis of 4-(4-(2-(3-methoxyphenyl)cyclopropyl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0379] Aqueous potassium hydroxide (50%, 25 mL) was added to a stirred suspension of nitrosomethylurea (4.0 g, 38.8 mmol) in ethyl ether (25 mL) at 0° C. The ether phase was separated and dried over potassium hydroxide. This resulting solution (25 mL) was added dropwise to a solution of (E)-4-(4-(3-methoxystyryl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (180 mg, 25.7 mmol) in tetrahydrofuran (50 mL), followed by palladium(II) acetate (38 mg, 0.17 mmol) and the mixture was stirred at 0° C. for 30 min. The reaction was then quenched with 2 mL of acetic acid, followed by the addition of water (100 mL) and the organic layer was separated. The aqueous layer was extracted with ethyl acetate (100 mL×2) and the combined organic layers were concentrated. The residue was subjected to silica gel column chromatography to give 4-(4-(2-(3-methoxyphenyl)cyclopropyl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (70 mg) as an off-white solid. 1 H NMR (400 MHz, CDCl3) δ 9.07 (d, J = 2.6Hz, 1H), 8.25 (dd, J = 4.7, 1.2Hz, 1H), 8.12 (ddd, J = 8.5, 2.7, 1.4Hz, 1H), 7.30 - 7.27 (m, 1H), 7.21 (t, J = 7.9Hz, 1H), 6.80 - 6.67 (m, 3H), 4.03 (dt, J = 12.0, 4.0Hz, 2H), 3.81 (s, 3H), 3.77 (s, 8H), 3.11 (dt, J = 9.0, 7.3Hz, 2H), 2.62 - 2.55 (m, 1H), 2.04 - 1.94 (m, 1H), 1.86 - 1.79 (m, 1H), 1.40 (ddd, J = 8.3, 6.1, 4.1Hz, 1H); LCMS (ESI) m / z: 430.2 [M+H] + . Synthesis of 4-(4-(2-(3-methoxyphenyl)pyrimidin-4-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (compound 123): [ka]

[0380] Step 1: Synthesis of 4-methoxy-2-(3-methoxyphenyl)pyrimidine.

[0381] A mixture of 2-chloro-4-methoxypyrimidine (2.88 g, 20.0 mmol), 3-methoxyphenylboronic acid (3.04 g, 20.0 mmol), 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex (816 mg, 1.0 mmol) and sodium carbonate (4.24 g, 40.0 mmol) in dioxane (100 mL) and water (4 mL) was stirred at 90° C. for 4 h under nitrogen atmosphere. The mixture was then poured into water and extracted with ethyl acetate (200 mL×2). The combined organic phase was concentrated and the residue was subjected to silica gel column chromatography (10% ethyl acetate in petroleum ether) to give 4-methoxy-2-(3-methoxyphenyl)pyrimidine (3.8 g) as a white solid. LCMS (ESI) m / z: 217.2 [M+H] + .

[0382] Step 2: Synthesis of 2-(3-methoxyphenyl)pyrimidin-4-ol.

[0383] A mixture of 4-methoxy-2-(3-methoxyphenyl)pyrimidine (3.7 g, 17.1 mmol) and hydrochloric acid (6N, 15 mL) was stirred at 100° C. for 2 h. The mixture was poured into water and extracted with dichloromethane (200 mL×2). The combined organic phase was dried and concentrated to give 2-(3-methoxyphenyl)pyrimidin-4-ol (2.2 g) as a white solid. LCMS (ESI) m / z: 203.1 [M+H] + .

[0384] Step 3: Synthesis of 4-chloro-2-(3-methoxyphenyl)pyrimidine.

[0385] A mixture of 2-(3-methoxyphenyl)pyrimidin-4-ol (2.0 g, 10.0 mmol) in phosphorus oxytrichloride (20 mL) was stirred at 120° C. for 2 h. The mixture was concentrated and the residue was dissolved in dichloromethane (200 mL) and poured into crushed ice. The organic layer was separated and the aqueous layer was extracted with dichloromethane (200 mL×3). The combined organic phase was concentrated and the residue was subjected to silica gel column chromatography (30% ethyl acetate in petroleum ether) to give 4-chloro-2-(3-methoxyphenyl)pyrimidine (1.8 g, 81%) as a grey solid. LCMS (ESI) m / z: 221.1 / 223.1 [M+H] + .

[0386] Step 4: Synthesis of 4-(4-(2-(3-methoxyphenyl)pyrimidin-4-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine.

[0387] A mixture of 4-chloro-2-(3-methoxyphenyl)pyrimidine (200 mg, 0.9 mmol), hexamethyldistannane (589 mg, 1.8 mmol) and bis(triphenylphosphine)palladium(II) chloride (64 mg, 0.09 mmol) in dioxane (15 mL) was stirred under nitrogen atmosphere at 100° C. for 2 h. The mixture was poured into dichloromethane (200 mL) and the organic phase was washed successively with saturated aqueous potassium fluoride (100 mL) and brine and concentrated to give 2-(3-methoxyphenyl)-4-(trimethylstannyl)pyrimidine (340 mg) as a brown oil. This oil was mixed with 4-(4-chloro-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (200 mg, 0.63 mmol), tetrakis(triphenylphosphine)palladium (104 mg, 0.09 mmol) in dioxane (15 mL) and stirred for an additional 2 h at 100° C. The mixture was concentrated and purified by silica gel column chromatography (25% methanol in dichloromethane) and further washed with methanol (20 mL) to give 4-(4-(2-(3-methoxyphenyl)pyrimidin-4-yl)-7-(pyridin-3-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl)morpholine (56.4 mg, 19%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 9.16 (s, 1H), 8.89 (d, J = 5.1Hz, 1H), 8.32 (d, J = 3.7Hz, 1H), 8.17 (t, J = 6.3Hz, 2H), 8.10 - 8.02 (m, 2H), 7.42 LCMS (ESI) m / z: 468.1 [M+H] + . Following the protocol described above, the following compounds were synthesized: [Table 18]

[0388] Synthesis of 4-(6-(1-methylpyrrolidin-3-yl)-9-phenyl-9H-purin-2-yl)morpholine (compound 126): [ka]

[0389] Step 1: Synthesis of 2,6-dichloro-9-phenyl-9H-purine.

[0390] To a solution of 2,6-dichloro-9H-purine (1.88 g, 10 mmol), phenylboronic acid (1.83 g, 15 mmol) in dichloromethane (50 mL) was added cupric acetate (900 mg, 5 mmol) and 1,10-phenanthroline (900 mg, 5 mmol), and the resulting mixture was stirred at room temperature under oxygen for 2 days. The mixture was filtered, and the filtrate was concentrated. The residue was subjected to flash chromatography eluting with 0-5% methanol in dichloromethane to give 2,6-dichloro-9-phenyl-9H-purine as a white solid (1.1 g, 42%). 1 H NMR (400 MHz, CDCl3) δ 8.40 (s, 1H), 7.60-7.60 (m, 2H), 7.56-7.46 (m, 2H), 7.48-7.44 (m, 1H); LCMS (ESI) m / z: 265.0 [M+H]+.

[0391] Step 2: Synthesis of tert-butyl 4-(2-chloro-9-phenyl-9H-purin-6-yl)-2,3-dihydro-1H-pyrrole-1-carboxylate.

[0392] To a solution of 2,6-dichloro-9-phenyl-9H-purine (132 mg, 0.5 mmol) in dioxane (5 mL) and water (1 mL), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-pyrrole-1-carboxylate (148 mg, 0.5 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (40 mg, 0.05 mmol) and sodium carbonate (159 mg, 1.5 mmol) were added at 25° C., and the resulting mixture was stirred under argon protection at 80° C. for 6 hours. It was cooled and the mixture was diluted with water (20 mL). The resulting precipitate was collected by filtration, washed with water (20 mL) and dried to give tert-butyl 4-(2-chloro-9-phenyl-9H-purin-6-yl)-2,3-dihydro-1H-pyrrole-1-carboxylate as a yellow solid (180 mg, 90%). LCMS (ESI) m / z: 342.1 [M-56+H] + .

[0393] Step 3: Synthesis of tert-butyl 4-(2-morpholino-9-phenyl-9H-purin-6-yl)-2,3-dihydro-1H-pyrrole-1-carboxylate.

[0394] To a mixture of tert-butyl 4-(2-chloro-9-phenyl-9H-purin-6-yl)-2,3-dihydro-1H-pyrrole-1-carboxylate (40 mg, 0.1 mmol) in N,N-dimethylacetamide (2 mL), morpholine (44 mg, 0.5 mmol) was added and the mixture was stirred at 100° C. for 16 h. It was then extracted with ethyl acetate (10 mL×3) and washed with water (10 mL×3). The combined organic layers were dried and concentrated. The residue was subjected to preparative TLC (UV254, silica, petroleum ether / ethyl acetate=1 / 1) to give tert-butyl 4-(2-morpholino-9-phenyl-9H-purin-6-yl)-2,3-dihydro-1H-pyrrole-1-carboxylate as a yellow solid (20 mg, 44%). LCMS (ESI) m / z: 449.1 [M+H] + .

[0395] Step 4: Synthesis of tert-butyl 3-(2-morpholino-9-phenyl-9H-purin-6-yl)pyrrolidine-1-carboxylate.

[0396] To a mixture of tert-butyl 4-(2-morpholino-9-phenyl-9H-purin-6-yl)-2,3-dihydro-1H-pyrrole-1-carboxylate (45 mg, 0.1 mmol) in methanol (5 mL) was added palladium on carbon (10%, 20 mg) and the suspension was stirred at room temperature under hydrogen for 2 h. The mixture was filtered and the filtrate was concentrated to give tert-butyl 3-(2-morpholino-9-phenyl-9H-purin-6-yl)pyrrolidine-1-carboxylate as a yellow solid. (45 mg, 99%). LCMS (ESI) m / z: 451.2 [M+H] + .

[0397] Step 5: Synthesis of 4-(9-phenyl-6-(pyrrolidin-3-yl)-9H-purin-2-yl)morpholine.

[0398] A mixture of tert-butyl 3-(2-morpholino-9-phenyl-9H-purin-6-yl)pyrrolidine-1-carboxylate (45 mg, 0.1 mmol) and hydrochloric acid / dioxane (4 M, 2 mL) in dichloromethane (5 mL) was stirred at room temperature for 2 hours. It was then diluted with 10 mL of dichloromethane and the mixture was washed with aqueous sodium bicarbonate (10 mL). The organic layer was concentrated to give 4-(9-phenyl-6-(pyrrolidin-3-yl)-9H-purin-2-yl)morpholine as a yellow solid (35 mg, 99%). LCMS (ESI) m / z: 351.2 [M+H] + .

[0399] Step 6: Synthesis of 4-(6-(1-methylpyrrolidin-3-yl)-9-phenyl-9H-purin-2-yl)morpholine.

[0400] To a solution of 4-(9-phenyl-6-(pyrrolidin-3-yl)-9H-purin-2-yl)morpholine (35 mg, 0.1 mmol) and formaldehyde (35%, 5 drops) in methanol (1 mL) and dichloroethane (2 mL), one drop of acetic acid was added and the mixture was stirred for 1 h. Sodium cyanoborohydride (31 mg, 0.5 mmol) was then added and the resulting mixture was stirred at room temperature for 16 h. The reaction was quenched with water (10 mL) and the mixture was extracted with dichloromethane (10 mL x 2). The organic phase was concentrated and the crude product was purified by preparative HPLC (BOSTON pHlex ODS 10um 21.2×250mm 120A. The mobile phase was acetonitrile / 0.1% ammonium bicarbonate) to give 4-(6-(1-methylpyrrolidin-3-yl)-9-phenyl-9H-purin-2-yl)morpholine (6.5 mg, 18%) as a white solid. 1 H NMR (400 MHz, CD3OD) δ 8.40 (s, 1H), 7.86 (d, J = 8.0Hz, 2H), 7.60 (t, J = 8.0Hz, 2H), 7.47 (t, J = 7.6Hz, 1H), 4.19 (pent, J = 8.4Hz, 1H), 3.88-3.77 (m, 8H), 3.26 (t. J = 9.2Hz, 1H), 3.08-2.83 (m, 3H), 2.52 (s, 3H), 2.44-2.37 (m, 2H); LCMS (ESI) m / z: 365.3 [M+H]+. Using a protocol similar to that described above, the following compounds were synthesized: [Table 19]

[0401] Synthesis of 4-(9-phenyl-6-(pyridin-4-yl)-9H-purin-2-yl)morpholine (compound 128): [ka]

[0402] Step 1: Synthesis of 2-chloro-9-phenyl-6-(pyridin-4-yl)-9H-purine.

[0403] To a solution of 2,6-dichloro-9-phenyl-9H-purine (264 mg, 1 mmol) in dioxane (10 mL) and water (2 mL) was added pyridin-4-ylboronic acid (123 mg, 1 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (81 mg, 0.1 mmol) and potassium carbonate (414 mg, 3 mmol) at 25°C, and the resulting mixture was stirred at 90°C for 16 h under argon protection. The mixture was extracted with ethyl acetate (20 mL x 3) and washed with water (20 mL). The organic layer was concentrated and the crude product was purified by preparative TLC (silica, UV254, ethyl acetate / petroleum ether = 3 / 1) to give 2-chloro-9-phenyl-6-(pyridin-4-yl)-9H-purine as a yellow solid (50 mg, 16%). LCMS(ESI)m / z:308.1[M+H] + (This step also produced 9-phenyl-2,6-di(pyridin-4-yl)-9H-purine (13 mg, 4%) as a by-product.)

[0404] Step 2: Synthesis of 4-(9-phenyl-6-(pyridin-4-yl)-9H-purin-2-yl)morpholine.

[0405] To a mixture of 2-chloro-9-phenyl-6-(pyridin-4-yl)-9H-purine (31 mg, 0.1 mmol) in N,N-dimethylacetamide (2 mL), morpholine (44 mg, 0.5 mmol) was added and stirred for 16 h at 100° C. The mixture was purified by preparative HPLC (BOSTON pHlex ODS 10 um 21.2×250 mm 120 A. The mobile phase was acetonitrile / 0.1% ammonium bicarbonate) to give 4-(9-phenyl-6-(pyridin-4-yl)-9H-purin-2-yl)morpholine as a yellow solid (26 mg, 72% yield). 1H NMR (400 MHz, DMSO-d6) δ 8.83 (d, J = 6.0Hz, 2H), 8.79 (s, 1H), 8.67 (d, J = 6.0Hz, 2H), 7.94 (d, J = 7.6Hz, 2H), 7.63 (t, J = 8.0Hz, 2H), 7.49 (t, J = 7.6Hz, 1H), 3.83-3.72 (m, 8H); LCMS (ESI) m / z: 359.2 [M+H]+. Synthesis of 2-morpholino-8-phenyl-4-(pyridin-3-ylmethoxy)-6H-pyrimido[5,4-b][1,4]oxazin-7(8H)-one (compound 129): [ka]

[0406] Step 1: Synthesis of methyl 2-(6-hydroxy-2-morpholino-4-oxo-1,4-dihydropyrimidin-5-yl)acetate.

[0407] To a solution of triethylethane-1,1,2-tricarboxylate (3 g, 12.18 mmol) and morpholine-4-carboximidamide hydrochloride (2 g, 12.18 mmol) in methanol (40 mL) was added sodium methanolate (30% solution in methanol, 6.7 mL, 34.35 mmol). After addition, the mixture was stirred at 80° C. for 17 h and concentrated. The crude product methyl 2-(6-hydroxy-2-morpholino-4-oxo-1,4-dihydropyrimidin-5-yl)acetate (3 g, 91.56%) was obtained as a brown solid, which was used in the next step without further purification. LCMS (ESI) m / z: 270.0 [M+H] + .

[0408] Step 2: Synthesis of methyl 2-(4,6-dichloro-2-morpholinopyrimidin-5-yl)acetate.

[0409] A mixture of methyl 2-(6-hydroxy-2-morpholino-4-oxo-1,4-dihydropyrimidin-5-yl)acetate (3 g, 11.15 mmol) and phosphorus oxychloride (20 mL) was stirred at 110° C. for 16 h and then concentrated. The residue was diluted with ethyl acetate / water (20 mL / 20 mL), the organic layer was separated, and the aqueous layer was extracted twice with ethyl acetate (20 mL). The combined organic phases were washed with brine (30 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by Combi-Flash® (Biotage, 40 g silica gel, eluted with 10% to 30% ethyl acetate in petroleum ether) to give methyl 2-(4,6-dichloro-2-morpholinopyrimidin-5-yl)acetate (1.3 g, 38.2%) as a white solid. LCMS (ESI) m / z: 306.1 [M+H] + .

[0410] Step 3: Synthesis of methyl 2-(4-chloro-2-morpholino-6-(pyridin-3-ylmethoxy)pyrimidin-5-yl)acetate.

[0411] To a solution of pyridin-3-ylmethanol (0.18 g, 1.65 mmol) in tetrahydrofuran (10 mL) was added sodium hydride (100 mg, 2.5 mmol) in portions and the mixture was stirred at 20° C. for 10 min. Then a solution of methyl 2-(4,6-dichloro-2-morpholinopyrimidin-5-yl)acetate (0.5 g, 1.64 mmol) in tetrahydrofuran (2 mL) was added slowly. After addition, the mixture was stirred at 20° C. for 2 h, then quenched with water (15 mL) and extracted with ethyl acetate (20 mL). The organic phase was dried over sodium sulfate, filtered and concentrated. The residue was purified by Combi-Flash® (Biotage, 40 g silica gel, eluted with 30% to 40% ethyl acetate in petroleum ether) to give methyl 2-(4-chloro-2-morpholino-6-(pyridin-3-ylmethoxy)pyrimidin-5-yl)acetate (0.3 g, 48.4%) as a white solid. LCMS (ESI) m / z: 379.2 [M+H] + .

[0412] Step 4: Synthesis of 2-morpholino-4-(pyridin-3-ylmethoxy)-7-m-tolyl-5H-pyrrolo[2,3-d]pyrimidin-6(7H)-one.

[0413] A mixture of methyl 2-(4-chloro-2-morpholino-6-(pyridin-3-ylmethoxy)pyrimidin-5-yl)acetate (0.16 g, 0.42 mmol), tris(dibenzylideneacetone)dipalladium (39 mg, 0.042 mmol), 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (40 mg, 0.084 mmol) and cesium carbonate (0.34 g, 1.06 mmol) in toluene (15 mL) was stirred at 90° C. for 3 h under nitrogen atmosphere. The reaction mixture was filtered and concentrated. The residue was purified by preparative HPLC to give 2-morpholino-4-(pyridin-3-ylmethoxy)-7-m-tolyl-5H-pyrrolo[2,3-d]pyrimidin-6(7H)-one (48 mg, 27.4%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.69 (d, J=1.6Hz, 1H), 8.55 (dd, J=4.8, 1.2hz, 1H), 7.88 (d, J = 8Hz, 1H), 7.46-7.34 (m, 2H), 7.27-7.16 (m, 3H), 5.49 (s, 2H), 3.64-3.51 (m, 10H), 2.35(s, 3H); LCMS (ESI) m / z: 417.9 [M+H] + . Synthesis of 2-morpholino-8-phenyl-4-(pyridin-3-ylmethoxy)-6H-pyrimido[5,4-b][1,4]oxazin-7(8H)-one (compound 130): [ka]

[0414] Step 1: Synthesis of dimethyl 2-(2-ethoxy-2-oxoethoxy)malonate.

[0415] A mixture of 1,3-dimethoxy-1,3-dioxopropane-2-diazonium (4 g, 25 mmol), ethyl 2-hydroxyacetate (1.2 mL, 12.5 mmol) and rhodium(II) acetate dimer (2 g, 4.5 mmol) in dichloromethane (40 mL) was stirred at 25° C. for 16 h. The reaction mixture was diluted with dichloromethane (20 mL) and filtered. The filtrate was concentrated and the residue was purified by flash chromatography (Biotage, 40 g silica gel, eluted with 30% to 60% ethyl acetate in petroleum ether) to give dimethyl 2-(2-ethoxy-2-oxoethoxy)malonate (3.3 g, 56%) as a colorless oil. LCMS (ESI) m / z: 235.1 [M+H] + .

[0416] Step 2: Synthesis of methyl 2-(6-hydroxy-2-morpholino-4-oxo-1,4-dihydropyrimidin-5-yloxy)acetate.

[0417] To a solution of dimethyl 2-(2-ethoxy-2-oxoethoxy)malonate (3 g, 12.82 mmol) and morpholine-4-carboximidamide hydrochloride (2.1 g, 12.82 mmol) in methanol (70 mL) was added sodium methanolate (30% solution in methanol, 7.5 mL, 38.46 mmol). After addition, the mixture was stirred at 80° C. for 17 h and concentrated to give methyl 2-(6-hydroxy-2-morpholino-4-oxo-1,4-dihydropyrimidin-5-yloxy)acetate (2.1 g, 57.5%) as a brown solid, which was used directly in the next step without further purification. LCMS (ESI) m / z: 286.1 [M+H] + .

[0418] Step 3: Synthesis of methyl 2-(4,6-dichloro-2-morpholinopyrimidin-5-yloxy)acetate.

[0419] A mixture of methyl 2-(6-hydroxy-2-morpholino-4-oxo-1,4-dihydropyrimidin-5-yloxy)acetate (2 g, 7 mmol), N,N-dimethylaniline (0.85 g, 7 mmol) and phosphorus oxychloride (15 mL) was stirred at 110° C. for 16 h. It was concentrated, the residue was diluted with ethyl acetate / water (20 mL / 20 mL), the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate (20 mL). The combined organic phase was washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. The residue was subjected to flash chromatography (Biotage, 40 g silica gel, eluted with 10% to 30% ethyl acetate in petroleum ether) to give methyl 2-(4,6-dichloro-2-morpholinopyrimidin-5-yloxy)acetate (0.85 g, 37.8%) as a yellow solid. LCMS(ESI)m / z:322.1[M+H] + .

[0420] Step 4: Synthesis of 2-(4-chloro-2-morpholino-6-(pyridin-3-ylmethoxy)pyrimidin-5-yloxy)acetic acid.

[0421] To a solution of pyridin-3-ylmethanol (68 mg, 0.62 mmol) in tetrahydrofuran (10 mL), sodium hydride (38 mg, 0.93 mmol) was added in portions and the mixture was stirred at 20° C. for 10 min. Then a solution of methyl 2-(4,6-dichloro-2-morpholinopyrimidin-5-yloxy)acetate (0.2 g, 0.62 mmol) in tetrahydrofuran (2 mL) was added slowly and the resulting mixture was stirred at 20° C. for 2 h. It was then quenched with water (15 mL) and extracted with ethyl acetate (20 mL). The aqueous phase was lyophilized to give crude 2-(4-chloro-2-morpholino-6-(pyridin-3-ylmethoxy)pyrimidin-5-yloxy)acetic acid (0.2 g, 87%) as a white solid, which was used directly in the next step without further purification. LCMS (ESI) m / z: 381.1 [M+H] + .

[0422] Step 5: Synthesis of 2-(4-chloro-2-morpholino-6-(pyridin-3-ylmethoxy)pyrimidin-5-yloxy)-N-phenylacetamide.

[0423] To a solution of 2-(4-chloro-2-morpholino-6-(pyridin-3-ylmethoxy)pyrimidin-5-yloxy)acetic acid (0.18 g, 0.47 mmol) and aniline (66 mg, 0.71 mmol) in N,N-dimethylformamide (15 mL) was added 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (0.32 g, 0.85 mmol) in portions, followed by N,N-diisopropylethylamine (0.18 g, 1.42 mmol). The resulting mixture was stirred at 20° C. for 2 h and then diluted with ethyl acetate / water (30 mL, 1:1). The layers were separated and the aqueous phase was extracted twice with ethyl acetate (20 mL). The combined organic phase was washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography (Biotage, 20 g silica gel, eluting with 7N ammonia in methanol:dichloromethane 1:10 15% to 20% in dichloromethane) to give 2-(4-chloro-2-morpholino-6-(pyridin-3-ylmethoxy)pyrimidin-5-yloxy)-N-phenylacetamide (0.11 g, 51%) as a yellow oil. LCMS (ESI) m / z: 456.1 [M+H] + .

[0424] Step 6: Synthesis of 2-morpholino-8-phenyl-4-(pyridin-3-ylmethoxy)-6H-pyrimido[5,4-b][1,4]oxazin-7(8H)-one.

[0425] A mixture of 2-(4-chloro-2-morpholino-6-(pyridin-3-ylmethoxy)pyrimidin-5-yloxy)-N-phenylacetamide (0.1 g, 0.22 mmol), tris(dibenzylideneacetone)dipalladium (20 mg, 0.022 mmol), 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (21 mg, 0.044 mmol) and cesium carbonate (0.18 g, 0.55 mmol) in toluene (10 mL) was stirred at 100° C. under nitrogen atmosphere for 16 h. It was filtered, concentrated and the residue was subjected to preparative HPLC to give 2-morpholino-8-phenyl-4-(pyridin-3-ylmethoxy)-6H-pyrimido[5,4-b][1,4]oxazin-7(8H)-one (5 mg, 5.4%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.69 (s, 1H), 8.56 (d, J=4.8Hz, 1H), 7.89 (d, J=8Hz, 1H), 7.52-7.38 (m, 4H), 7.28 (d, J=7.2Hz, 2H), 5.46 (s, 2H), 4.77 (s, 2H), 3.54-3.57 (m, 4H), 3.34-3.27 (m, 4H); LCMS (ESI) m / z: 420.0 [M+H] + . Synthesis of 4-(1-phenyl-6-(pyridin-3-ylmethoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)morpholine (compound 131): [ka]

[0426] Step 1: Synthesis of 4,6-dichloro-1-phenyl-1H-pyrazolo[3,4-d]pyrimidine.

[0427] To a stirred solution of 2,4,6-trichloropyrimidine-5-carbaldehyde (630 mg, 3 mmol) in ethanol (20 mL) at -78°C, phenylhydrazine (324 mg, 3 mmol) and triethylamine (910 mg, 9 mmol) were added dropwise in that order. The resulting mixture was stirred at -78°C for 0.5 h and at 0°C for 2 h. The mixture was then quenched with water (20 mL) and the resulting precipitate was collected by filtration and dried to give 4,6-dichloro-1-phenyl-1H-pyrazolo[3,4-d]pyrimidine as a white solid (790 mg, 99%). LCMS (ESI) m / z: 265.0 [M+H] + .

[0428] Step 2: Synthesis of 4-(6-chloro-1-phenyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)morpholine.

[0429] To a mixture of 4,6-dichloro-1-phenyl-1H-pyrazolo[3,4-d]pyrimidine (792 mg, 3 mmol) in dichloromethane (10 mL) was added morpholine (520 mg, 6 mmol) and DIPEA (774 mg, 6 mmol) at 0° C., and the resulting mixture was stirred at room temperature for 16 h. The mixture was concentrated and purified by column chromatography eluting with 0-30% ethyl acetate in petroleum ether to give 4-(6-chloro-1-phenyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)morpholine as a yellow solid (800 mg, 85%). LCMS (ESI) m / z: 316.0 [M+H] + .

[0430] Step 3: Synthesis of 4-(1-phenyl-6-(pyridin-3-ylmethoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)morpholine.

[0431] To a mixture of pyridin-3-ylmethanol (218 mg, 2 mmol) in tetrahydrofuran (10 mL) was added sodium hydride (120 mg, 3 mmol) followed by 4-(6-chloro-1-phenyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)morpholine (315 mg, 1 mmol) at 0° C. and the resulting mixture was stirred at room temperature for 16 hours. The reaction was quenched with water (10 mL) and the resulting precipitate was filtered off and dried. The crude product thus obtained was purified by preparative HPLC (BOSTON pHlex ODS 10 um 21.2×250 mm 120A. The mobile phase was acetonitrile / 0.1% formic acid) to give 4-(1-phenyl-6-(pyridin-3-ylmethoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)morpholine as a yellow solid. (75 mg, 19%). 1 H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 1.6Hz, 1H), 8.54 (dd, J = 4.8, 1.6Hz, 1H), 8.47 (s, 1H), 8.11 (dd, J = 4.8, 0.8Hz, 2H), 7.90-7.87 (m, 1H), 7.56-7.52 (m, 2H), 7.43-7.32 (m, 2H), 5.44 (s, 2H), 3.92 (t, J = 4.8Hz, 4H), 3.75 (t, J = 4.8Hz, 4H);LCMS (ESI) m / z: 389.2 [M+H]+. Synthesis of 4-(1-phenyl-4-(pyridin-3-ylmethoxy)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)morpholine (compound 132): [ka]

[0432] Step 1: Synthesis of 6-chloro-1-phenyl-4-(pyridin-3-ylmethoxy)-1H-pyrazolo[3,4-d]pyrimidine.

[0433] To a solution of pyridin-3-ylmethanol (109 mg, 1 mmol) in tetrahydrofuran (10 mL) was added sodium hydride (60 mg, 1.5 mmol, 60%) at 0° C., followed by 4,6-dichloro-1-phenyl-1H-pyrazolo[3,4-d]pyrimidine (264 mg, 1 mmol), and the resulting mixture was stirred at room temperature for 16 h. The reaction was quenched with water (10 mL), and the mixture was extracted with ethyl acetate (20 mL×2). The organic layer was dried and concentrated to give 6-chloro-1-phenyl-4-(pyridin-3-ylmethoxy)-1H-pyrazolo[3,4-d]pyrimidine as a yellow solid. (250 mg, 74%). LCMS (ESI) m / z: 338.0 [M+H] + .

[0434] Step 2: Synthesis of 4-(1-phenyl-4-(pyridin-3-ylmethoxy)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)morpholine.

[0435] To a mixture of 6-chloro-1-phenyl-4-(pyridin-3-ylmethoxy)-1H-pyrazolo[3,4-d]pyrimidine (170 mg, 0.5 mmol) in dichloromethane (10 mL) was added morpholine (82 mg, 1 mmol) at 0° C., followed by DIPEA (129 mg, 1 mmol), and the resulting mixture was stirred at room temperature for 16 hours. It was then concentrated and the resulting crude product was purified by preparative HPLC (BOSTON pHlex ODS 10 um 21.2×250 mm 120A. The mobile phase was acetonitrile / 0.1% ammonium bicarbonate) to give 4-(1-phenyl-4-(pyridin-3-ylmethoxy)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)morpholine as a yellow solid. (40 mg, 21%). 1H NMR (400 MHz, DMSO-d6) δ 8.76 (d, J = 4.6Hz, 1H), 8.58 (dd, J = 4.8Hz, 1H), 8.20-8.18 (m, 3H), 7.95 (d, J = 8Hz, 1H), 7.53 (t,J = 8Hz, 2H), 7.45 (dd, J = 7.6, 4.8hz, 1H), 7.30 (t, J = 6.8Hz, 1H), 5.63 (s, 2H), 3.83 (t, J = 4.4Hz, 4H), 3.70 (t, J = 4.8Hz, 4H); LCMS (ESI) m / z: 389.1 [M+H]+. Synthesis of 4-(3-phenyl-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl)morpholine (compound 133): [ka]

[0436] Step 1: Synthesis of 2-chloro-4-methyl-6-(pyridin-4-yl)pyrimidin-5-amine.

[0437] A mixture of 2,4-dichloro-6-methylpyrimidin-5-amine (9 g, 51 mmol), pyridin-4-ylboronic acid (6.21 g, 51 mmol), potassium carbonate (17.5 g, 126 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (3.7 g, 5.1 mmol) in dioxane / water (150 mL / 25 mL) was stirred at 80° C. under nitrogen atmosphere for 4 h. The reaction mixture was concentrated and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=20:1) to give the target product as a yellow solid (6.4 g, 57%). LCMS (ESI) m / z: 221.0 [M+H] + .

[0438] Step 2: Synthesis of 5-chloro-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidine.

[0439] To a mixture of 2-chloro-4-methyl-6-(pyridin-4-yl)pyrimidin-5-amine (4.55 g, 20.6 mmol), acetic anhydride (4.42 g, 43.3 mmol), potassium acetate (4.05 g, 41.2 mmol) and acetic acid (4.33 g, 72.2 mmol) in toluene (100 mL), tert-butyl nitrite (2.66 g, 25.8 mmol) was added at room temperature and the reaction was stirred at 120° C. for 2 hours. It was concentrated and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=15:1) to give the target product as a yellow solid (2.3 g, 48%). LCMS (ESI) m / z: 232.0 [M+H] + .

[0440] Step 3: Synthesis of 3-bromo-5-chloro-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidine.

[0441] A mixture of 5-chloro-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidine (2.1 g, 9.07 mmol) and N-bromosuccinimide (2.3 g, 9.97 mmol) in N,N-dimethylformamide (25 mL) was stirred at 25° C. for 2 h. The mixture was diluted with water and extracted with ethyl acetate (100 m×3). The combined organic layers were concentrated and the residue was subjected to silica gel column chromatography (dichloromethane:methanol=10:1) to give the target product as a yellow solid (1.6 g, 57%). LCMS (ESI) m / z: 310.0 [M+H] + .

[0442] Step 4: Synthesis of 4-(3-bromo-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl)morpholine.

[0443] A mixture of 3-bromo-5-chloro-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidine (1.6 g, 5.15 mmol) and morpholine (4.49 g, 51.52 mmol) in 1-methyl-2-pyrrolidinone (20 mL) was stirred at 100° C. for 8 h. Water (100 mL) was added to the mixture and the resulting precipitate was filtered and dried to give the target product as a yellow solid (1.1 g, 59.11%). LCMS (ESI) m / z: 361.0 [M+H] + .

[0444] Step 5: Synthesis of 4-(3-phenyl-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl)morpholine.

[0445] A mixture of 4-(3-bromo-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl)morpholine (0.08 g, 0.22 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.016 g, 0.022 mmol), cesium carbonate (0.217 g, 0.66 mmol) and phenylboronic acid (0.054 g, 0.44 mmol) in dioxane / water (3 mL / 0.5 mL) was stirred at 90° C. for 4 h. It was then concentrated and the residue was subjected to preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The mobile phase was acetonitrile / 10 mM formic acid in water) to give the target product as a yellow solid (11.8 mg, 14.87%). 1 H NMR (400 MHz, DMSO-d6) δ 14.11 (s, 1H), 8.88 (s, 2H), 8.44 (d, J = 7.3Hz, 2H), 8.26 (s, 2H), 7.53 (d, J = 7.4Hz, 2H), 7.40 (d, J = 6.2Hz, 1H), 3.87 (s, 4H), 3.78 (s, 4H); LCMS (ESI) m / z: 358.8 [M] + . Synthesis of 4-(3-(3-(1H-pyrazol-1-yl)phenyl)-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl)morpholine (compound 134): [ka]

[0446] A mixture of 4-(3-bromo-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl)morpholine (100 mg, 0.28 mmol), 1-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazole (48 mg, 0.32 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (20 mg, 0.0028 mmol) and cesium carbonate (209 mg, 0.84 mmol) in dioxane (5 mL) and water (0.5 mL) was stirred at 110° C. for 16 h under nitrogen atmosphere. Water was then added and the mixture was extracted with ethyl acetate (50 mL×3). The organic layer was dried and concentrated, and the resulting crude product was purified by preparative TLC (petroleum ether:ethyl acetate=50:1 to 10:1) to give 4-(3-(3-(1H-pyrazol-1-yl)phenyl)-7-(pyridin-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl)morpholine (14.8 mg, 24%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 14.00 (s, 1H), 9.00 (s, 1H), 8.88 (dd, J = 4.5, 1.6Hz, 2H), 8.56 (s, 1H), 8.41 (s, 1H), 8.17 (s, 1H), 7.88 -7.76 (m, 2H), 7.66 (t, J = 8.1Hz, 1H), 6.60 (s, 1H), 3.91 (s, 4H), 3.86 - 3.68 (m, 4H); LCMS (ESI) m / z: 425.1 [M+H] + . Synthesis of 4-(7-phenyl-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (compound 135): [ka]

[0447] Step 1: Synthesis of 2-chloro-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine.

[0448] To a solution of 2,4-dichloro-5H-pyrrolo[3,2-d]pyrimidine (15 g, 77 mmol) in dioxane / water (200 mL / 40 mL) was added pyridin-4-ylboronic acid (5.8 g, 77 mmol), potassium carbonate (21.3 g, 154 mmol) and [1,1'bis(diphenylphosphino)ferrocene]dichloro-palladium(II) (5.6 g, 7.7 mmol) at 25° C., and the resulting mixture was stirred at 85° C. under argon protection for 2 hours. The mixture was then filtered and the filtrate was concentrated to give the target product as a dark solid (15 g, 84%). LCMS (ESI) m / z: 231.1 [M+H] + .

[0449] Step 2: Synthesis of 7-bromo-2-chloro-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine.

[0450] A mixture of 2-chloro-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine (3.0 g, 13 mmol) and N-bromosuccinimide (2.3 g, 13 mmol) in N,N-dimethylformamide (30 mL) was stirred at 25° C. for 2 h. Methanol (100 mL) was added to the mixture, which was filtered, and the filtrate was concentrated. The resulting residue was subjected to silica gel column chromatography (petroleum ether:ethyl acetate=1:2) to give the target product as a yellow solid (2 g, 50%). LCMS (ESI) m / z: 309.2 [M+H] + .

[0451] Step 3: Synthesis of 4-(7-bromo-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine.

[0452] A mixture of 7-bromo-2-chloro-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine (0.3 g, 0.97 mmol) and morpholine (0.5 g, 5.8 mmol) in NMP (3 mL) was stirred at 110° C. for 5 h. It was cooled to room temperature and quenched with water (15 mL). The mixture was extracted with ethyl acetate (15 mL×3), the organic layer was concentrated and subjected to preparative TLC (dichloromethane:acetic ester=1:1) to give the target product as a yellow solid (0.08 g, 23%). LCMS (ESI) m / z: 360.1 [M+H] + .

[0453] Step 4: Synthesis of 4-(7-phenyl-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine.

[0454] A mixture of 4-(7-bromo-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (0.07 g, 0.19 mmol), [1,1'bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.015 g, 0.02 mmol), cesium carbonate (0.19 g, 0.58 mmol) and phenylboronic acid (0.05 g, 0.39 mmol) in dioxane / water (3 mL / 0.5 mL) was stirred at 90° C. for 2 h. The mixture was concentrated and the resulting residue was subjected to preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The mobile phase was acetonitrile / 10 mM formic acid in water) to give the target product as a yellow solid (0.0198 g, 29%). 1H NMR (400 MHz, DMSO-d6) δ 11.96 (s, 1H), 8.81 (d, J = 5.3Hz, 2H), 8.38 (bs, 1H), 8.32 (s, 1H), 8.26 (d, J = 7.8Hz, 2H), 8.05 (d, J = 5.1Hz, LCMS (ESI) m / z: 358.2 [M+H] + . Synthesis of 4-(7-(pyridin-2-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (compound 136): [ka]

[0455] A mixture of 4-(7-bromo-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (60 mg, 0.16 mmol), 2-(trimethylstannyl)pyridine (48.2 mg, 0.2 mmol) and tetratriphenylphosphonium palladium (18 mg, 0.016 mmol) in dioxane (5 mL) was stirred under nitrogen protection at 100° C. for 16 hours. The resulting crude product was purified by flash chromatography (petroleum ether / ethyl acetate 20:1→10:1→5:1) to give 4-(7-(pyridin-2-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (4.4 mg, 8%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 12.08 (s, 1H), 8.82 (d, J = 5.9Hz, 2H), 8.66 (d, J = 7.9Hz, 1H), 8.56 (d, J = 4.6Hz, 1H), 8.38 (d, J = 3.2Hz, LCMS (ESI) m / z: 358.8 [M+H]+ Synthesis of 4-(4-(pyridin-4-yl)-7-(pyrimidin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (compound 137): [ka]

[0456] To a solution of 4-(7-bromo-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (100 mg, 0.28 mmol) in dioxane (10 mL), 4-(tributylstannyl)pyrimidine (121 mg, 0.33 mmol), lithium chloride (35 mg, 0.84 mmol) and bis(tri-tert-butylphosphine)palladium (15 mg, 0.028 mmol) were added at 25° C., and the resulting mixture was stirred at 110° C. for 3 hours under argon protection. The mixture was then concentrated and the residue was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. Mobile phase was acetonitrile / 10 mM aqueous trifluoroacetic acid) to give 4-(4-(pyridin-4-yl)-7-(pyrimidin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (23 mg, 23.1%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.04 (s, 1H), 8.80 (d, J = 5.9Hz, 2H), 8.74 (d, J = 5.4Hz, 1H), 8.60 (d, J = 5.4Hz, 1H), 8.53 (s, 1H), 8.11 (d, J = 5.6Hz, 2H), 3.84 (d, J = 5.2Hz, 4H), 3.78 (d, J = 4.5Hz, 4H). Synthesis of 4-(7-(4-cyclopropylpyrimidin-2-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (compound 138): [ka]

[0457] Step 1: Synthesis of 2-chloro-4-cyclopropylpyrimidine.

[0458] To a solution of 2,4-dichloropyrimidine (1.03 g, 6.92 mmol) in dioxane (15 mL) and water (3 mL) was added cyclopropylboronic acid (722 mg, 8.4 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (511.7 mg, 0.7 mmol) and potassium carbonate (2.9 g, 21.0 mmol) at 25°C, and the mixture was stirred at 90°C for 2 h under nitrogen protection. It was then extracted with ethyl acetate (20 mL x 2) and washed with water (10 mL x 2). The organic layer was dried over sodium sulfate and concentrated. The residue was subjected to silica gel column chromatography (35% ethyl acetate in petroleum ether) to give 2-chloro-4-cyclopropylpyrimidine as a colorless oil (900 mg, 83.5%). LCMS (ESI) m / z: 154.9 [M+H] + .

[0459] Step 2: Synthesis of 4-cyclopropyl-2-(trimethylstannyl)pyrimidine.

[0460] To a solution of 2-chloro-4-cyclopropylpyrimidine (308 mg, 2.0 mmol) in dioxane (10 mL) at 25° C., 1,1,1,2,2,2-hexamethyldistannane (1.31 mg, 4.0 mmol) and bis(triphenylphosphine)palladium(II) chloride (140.2 mg, 0.2 mmol) were added and the reaction was stirred at 100° C. for 2 h under nitrogen protection. Aqueous potassium fluoride (50 mL) was added to the mixture, which was filtered. The filtrate was extracted with dichloromethane (30 mL×3) and the organics were concentrated to give 4-cyclopropyl-2-(trimethylstannyl)pyrimidine as a yellow oil (500 mg, 88.0%). LCMS (ESI) m / z: 285.0 [M+H] + .

[0461] Step 3: Synthesis of 4-(7-(4-cyclopropylpyrimidin-2-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine.

[0462] To a solution of 4-(7-bromo-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (150 mg, 0.4 mmol) in dioxane (10 mL), 4-cyclopropyl-2-(trimethylstannyl)pyrimidine (255 mg, 0.8 mmol) and bis(triphenylphosphine)palladium(II) chloride (52 mg, 0.04 mmol) were added at 25° C., and the resulting mixture was stirred at 100° C. for 2 hours under nitrogen protection. The mixture was then extracted with dichloromethane (20 mL×2), and the combined organic layers were washed with water (10 mL×2), dried over sodium sulfate, and concentrated. The residue was subjected to preparative HPLC (BOSTON pHlex ODS 10um 21.2×250mm 120A. The mobile phase was DMSO / 0.1% ammonium bicarbonate) to give 4-(7-(4-cyclopropylpyrimidin-2-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine as a yellow solid (9.0 mg, 5.6%). 1 H NMR (400 MHz, DMSO-d6) δ 12.11 (s, 1H), 8.81 (d, J = 5.9Hz, 2H), 8.55 (d, J = 5.1Hz, 1H), 8.36 (s, 1H), 8.29 (s, 1H), 8.01 (d, J = 6.0Hz, 2H), 7.19 (d, J = 5.1Hz, 1H), 3.84 (d, J = 4.9Hz, 4H), 3.75 (d, J = 4.7Hz, 4H), 2.10 (pent, J = 4.2Hz, 1H), 1.24 (d, J = 4.0Hz, 2H), 1.07 (dd, J = 7.8, 3.2Hz, 2H); LCMS (ESI) m / z: 399.9 [M] + . Synthesis of 4-(7-(6-methoxypyridin-2-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (compound 139): [ka]

[0463] To a solution of 4-(7-bromo-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (100 mg, 0.28 mmol) in dioxane / water (10 mL / 1 mL), (6-methoxypyridin-2-yl)boronic acid (51 mg, 0.33 mmol), cesium carbonate (273 mg, 0.84 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (23 mg, 0.028 mmol) were added at 25°C, and the resulting mixture was stirred at 110°C for 3 hours under argon protection. The mixture was then concentrated and the residue was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. Mobile phase was acetonitrile / 10 mM aqueous trifluoroacetic acid) to give 4-(7-(6-methoxypyridin-2-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (21 mg, 19.5%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 12.04 (s,1H), 8.83(d, J = 5.7Hz, 2H), 8.34 (d, J = 3.3Hz, 1H), 8.23 ​​(d, J = 7.0Hz, 1H), 8.04 (d, J = 5.8Hz, 2H), 7.76 (t, J = 8Hz, 1H), 6.62 (d, J = 7.8Hz, 1H), 3.96 (s, 3H), 3.82 (d, J = 5.0Hz, 4H), 3.77 (d, J = 5.0Hz, 4H); LCMS (ESI) m / z: 388.8 [M]+. Following the protocol described above, the following compounds were synthesized: [Table 20-1] [Table 20-2]

[0464] Synthesis of 4-(7-(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (compound 146): [ka]

[0465] Step 1: Synthesis of tert-butyl 7-bromo-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate.

[0466] A mixture of 4-(7-bromo-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (400 mg, 1.1 mmol), di-tert-butyl dicarbonate (285 mg, 1.32 mmol) and N,N-dimethylpyridin-4-amine (14 mg, 0.11 mmol) in tetrahydrofuran (20 mL) was stirred at 15° C. for 2 h. The resulting precipitate was collected by filtration to give tert-butyl 7-bromo-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (460 mg, 91.1%) as a pale yellow solid. LCMS (ESI) m / z: 459.8 / 461.8 [M+H] + .

[0467] Step 2: Synthesis of 5-(tert-butoxycarbonyl)-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-7-ylboronic acid.

[0468] A mixture of tert-butyl 7-bromo-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (400 mg, 0.84 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (426 mg, 1.68 mmol), 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex (32 mg, 0.04 mmol) and cesium carbonate (546 mg, 1.68 mmol) in dioxane (10 mL) and water (1.5 mL) was stirred at 100° C. under nitrogen atmosphere for 16 hours. The mixture was poured into water and extracted with ethyl acetate (150 mL×2). The combined organic phase was concentrated to give 5-(tert-butoxycarbonyl)-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-7-ylboronic acid (1.8 g, crude) as a brown oil, which was used in the next step without further purification. LCMS (ESI) m / z: 425.9 [M+H] + .

[0469] Step 3: Synthesis of tert-butyl 7-(5-(tert-butoxycarbonyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate.

[0470] A mixture of tert-butyl 2-morpholino-4-(pyridin-4-yl)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (1.8 g, crude, from previous step), tert-butyl 3-bromo-6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)-carboxylate (100 mg, 0.33 mmol), 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex (27 mg, 0.033 mmol) and cesium carbonate (214 mg, 0.66 mmol) in dioxane (10 mL) and water (2 mL) was stirred at 100 °C under nitrogen atmosphere for 2 h. The mixture was poured into water and extracted with ethyl acetate (150 mL x 2). The combined organic phase was concentrated and the residue was subjected to silica gel column chromatography (10% dichloromethane in methanol) and preparative HPLC (column Xbridge 21.2 x 250 mm C18, 10 um, mobile phase A: water (10 mmol / L ammonium bicarbonate) B: acetonitrile) successively to give tert-butyl 7-(5-(tert-butoxycarbonyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (150 mg, 75%) as a yellow solid. LCMS (ESI) m / z: 603.2 [M+H] + .

[0471] Step 4: Synthesis of 4-(4-(pyridin-4-yl)-7-(4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine.

[0472] A mixture of tert-butyl 7-(5-(tert-butoxycarbonyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (70 mg, 0.11 mmol) and hydrochloric acid (4 M in dioxane, 2 mL) in dichloromethane (10 mL) was stirred for 2 h at 30° C. The mixture was neutralized with ammonia (7.0 M in methanol, 20 mL) and concentrated. The residue was subjected to silica gel column chromatography (30% dichloromethane in methanol) to give 4-(4-(pyridin-4-yl)-7-(4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (30 mg, 67%) as a pale yellow solid. LCMS (ESI) m / z: 402.9 [M+H] + .

[0473] Step 5: Synthesis of 4-(7-(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine.

[0474] A mixture of 4-(4-(pyridin-4-yl)-7-(4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (25 mg, 0.062), formaldehyde (40% in water, 2 mL), acetic acid (0.05 mL) and methanol (5 mL) was stirred at 20° C. for 0.5 h followed by the addition of sodium cyanoborohydride (20 mg, 0.31 mmol). The mixture was stirred at 20° C. for an additional 0.5 h and concentrated. The residue was subjected to preparative HPLC (column Xbridge 21.2×250 mm C18, 10 um, mobile phase A: water (10 mmol / L ammonium bicarbonate) B: acetonitrile) to give 4-(7-(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (17.9 mg, 69.3%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 8.82 (d, J = 6.0Hz, 2H), 8.45 (s, 1H), 8.05 (s, 1H), 7.86 (dd, J = 4.5, 1.4Hz, 2H), 7.43 (d, J = 2.8Hz, 1H), 4.30 LCMS (ESI) m / z: 416.9 [M] + . Synthesis of 4-(7-(5-(cyclopropylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (compound 147): [ka]

[0475] A mixture of 4-(4-(pyridin-4-yl)-7-(4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (15 mg, 0.037 mmol), cyclopropanecarbaldehyde (13 mg, 0.18 mmol), acetic acid (0.05 mL) and methanol (5 mL) was stirred at 20° C. for 30 min followed by the addition of sodium cyanoborohydride (20 mg, 0.31 mmol). The mixture was stirred at 20° C. for an additional 30 min and concentrated. The resulting crude product was purified by preparative HPLC (column Xbridge 21.2×250 mm C18, 10 um, mobile phase A: water (10 mmol / L ammonium bicarbonate) B: acetonitrile) to give 4-(7-(5-(cyclopropylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (12.1 mg, 69.3%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 11.52 (s, 1H), 8.63 (d, J = 4Hz, 2H), 7.86 (dd, J = 6.7, 5.2Hz, 3H), 7.58 (d, J = 2.5Hz, 1H), 3.98 (t, J = 5.2Hz, 2H), 3.81 (s, 2H), 3.58 (s, 8H), 2.83 (t, J = 5.3Hz, 2H), 2.31 (d, J = 6.6Hz, 2H), 0.77 (s, 1H), 0.34 (q, J = 5.4Hz, 2H), -0.01 (d, J = 4.3Hz, 2H); LCMS (ESI) m / z: 456.8 [M] + . Synthesis of 4-(7-(6-methoxypyridazin-3-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (compound 148): [ka]

[0476] Step 1: Synthesis of tert-butyl 7-(6-methoxypyridazin-3-yl)-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate.

[0477] A solution of (tert-butyl 2-morpholino-4-(pyridin-4-yl)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (100 mg, 0.28 mmol), 3-bromo-6-methoxypyridazine (31 mg, 0.34 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (20 mg, 0.028 mmol) and cesium carbonate (270 mg, A mixture of 0.84 mmol) was stirred at 120° C. under nitrogen atmosphere for 16 h. The mixture was extracted with ethyl acetate (50 mL×3), and the organic layer was dried and concentrated. The resulting crude product was purified by preparative TLC (petroleum ether:ethyl acetate 50:1 to 10:1) to give tert-butyl 7-(6-methoxypyridazin-3-yl)-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (54 mg, 42%) as a yellow solid; LCMS (ESI) m / z: 489.7 [M+H] + .

[0478] Step 2: Synthesis of 4-(7-(6-methoxypyridazin-3-yl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine.

[0479] To a solution of tert-butyl 7-(6-methoxypyridazin-3-yl)-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (54 mg, 0.11 mmol) in acetonitrile (10 mL) was added hydrochloric acid (3 mol / L in methanol, 5 mL). The mixture was stirred at 25° C. for 2 hours and concentrated. The residue was purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The mobile phase was acetonitrile / 10 mM formic acid in water) to give the target product as a yellow solid (13.1 mg, 30%). 1HNMR (400MHz, DMSO-d6) δ 12.21 (s, 1H), 8.83-8.80 (m, 3H), 8.46 (s, 1H), 8.05 (d, J = 5.7Hz, 2H), 7.33 (d, J = 9.1Hz, 1H), 4.05 (s, 3H), 3.82 (s, 4H), 3.77 (s, 4H). LCMS (ESI) m / z: 389.8 [M+H] + . Synthesis of 2-(7-(3-(1H-pyrazol-1-yl)phenyl)-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-5-yl)ethan-1-ol (compound 149): [ka]

[0480] Step 1: Synthesis of 4-(7-(3-(1H-pyrazol-1-yl)phenyl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine.

[0481] A mixture of 4-(7-bromo-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (100 mg, 0.28 mmol), 1-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazole (31 mg, 0.34 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (20 mg, 0.028 mmol) and cesium carbonate (270 mg, 0.84 mmol) in dioxane (5 mL) and water (0.5 mL) was stirred at 120° C. for 16 h under a nitrogen atmosphere. The mixture was extracted with ethyl acetate (50 mL x 3) and the combined organic layers were dried, concentrated and purified by preparative TLC (petroleum ether:ethyl acetate 50:1 to 5:1) to give 4-(7-(3-(1H-pyrazol-1-yl)phenyl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (94 mg, 72%) as a yellow solid. LCMS (ESI) m / z: 424.8 [M+H] +

[0482] Step 2: Synthesis of 2-(7-(3-(1H-pyrazol-1-yl)phenyl)-2-morpholino-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-5-yl)ethan-1-ol.

[0483] To a solution of 4-(7-(3-(1H-pyrazol-1-yl)phenyl)-4-(pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (94 mg, 0.21 mmol) and 2-bromoethanol (0.31 mmol) in dimethylsulfoxide (10 mL) was added potassium hydroxide (31 mg, 0.63 mmol) at 25° C. and the mixture was stirred at 70° C. for 16 hours. The reaction was quenched with water and extracted with ethyl acetate (50 mL×3). The organic layer was dried, concentrated and purified by preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The mobile phase was acetonitrile / 10 mM formic acid in water) to give the target product as a brown solid (9.2 mg, 10%). 1H NMR (400 MHz, DMSO-d6) δ 9.58 (s, 2H), 8.99 (d, J = 6.4Hz, 2H), 8.73 (s, 1H), 8.47 - 8.45 (m, 2H), 8.14 (s, 1H), 7.75 (s, 1H), 7.41 (d, J = 4.6Hz, 2H), 6.56 (s, 1H), 5.30 (s, 1H), 4.66 (s, 2H), 3.90 (s, 2H), 3.82 (s, 8H); LCMS (ESI) m / z: 467.7 [M+H]+. Synthesis of 4-(7-(3-(1H-pyrazol-1-yl)phenyl)-4-(pyridazin-3-ylmethoxy)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (compound 150): [ka]

[0484] Step 1: Synthesis of tert-butyl 7-bromo-2-morpholino-4-(pyridazin-3-ylmethoxy)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate.

[0485] A solution of (4-(7-bromo-4-(pyridazin-3-ylmethoxy)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine (100 mg, 0.26 mmol), di-tert-butyl dicarbonate (67 mg, 0.31 mmol) and 4-dimethylaminopyridine (10 mg, 0.05 mmol) in tetrahydrofuran (5 mL) was stirred at room temperature under a nitrogen atmosphere for 4 hours. The resulting mixture was cooled to room temperature and cooled to room temperature. The mixture was extracted with ethyl acetate (50 mL x 3) and the combined organic layers were dried and concentrated. The residue was subjected to preparative TLC (petroleum ether:ethyl acetate 50:1 to 10:1) to give tert-butyl 7-bromo-2-morpholino-4-(pyridazin-3-ylmethoxy)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (110 mg, 92%) as a yellow oil. LCMS (ESI) m / z: 491.0 [M+H] +

[0486] Step 2: Synthesis of tert-butyl 7-(3-(1H-pyrazol-1-yl)phenyl)-2-morpholino-4-(pyridazin-3-ylmethoxy)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate.

[0487] A mixture of tert-butyl 7-bromo-2-morpholino-4-(pyridazin-3-ylmethoxy)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (100 mg, 0.2 mmol), 1-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazole (66 mg, 0.22 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (20 mg, 0.02 mmol) and cesium carbonate (195 mg, 0.6 mmol) in dioxane (5 mL) and water (0.5 mL) was stirred at 100° C. under nitrogen atmosphere for 16 h. The mixture was extracted with ethyl acetate (50 mL×3) and the combined organic phase was dried and concentrated. The residue was purified by preparative TLC (petroleum ether:ethyl acetate 50:1 to 10:1) to give tert-butyl 7-(3-(1H-pyrazol-1-yl)phenyl)-2-morpholino-4-(pyridazin-3-ylmethoxy)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (100 mg, 82%) as a yellow solid. LCMS (ESI) m / z: 555.1 [M+H] +

[0488] Step 3: Synthesis of 4-(7-(3-(1H-pyrazol-1-yl)phenyl)-4-(pyridazin-3-ylmethoxy)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)morpholine.

[0489] To a solution of tert-butyl 7-(3-(1H-pyrazol-1-yl)phenyl)-2-morpholino-4-(pyridazin-3-ylmethoxy)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (100 mg, 0.11 mmol) in acetonitrile (10 mL) was added hydrochloric acid (3 mol / L in methanol, 5 mL). The resulting mixture was stirred at 25° C. for 2 hours and concentrated. The residue was subjected to preparative HPLC (SunFire C18, 4.6×50 mm, 3.5 um column Xbridge C18 3.5 μm 4.6×50 mm column. The mobile phase was acetonitrile / 10 mM formic acid in water) to give the target product as a white solid (40.6 mg, 50%). 1H NMR (400 MHz, DMSO-d6) δ 9.22 (dd, J = 4.9, 1.6Hz, 1H), 8.75 (s, 1H), 8.51 (d, J = 2.5Hz, 1H), 8.19 (s, 1H), 8.15 (d, J = 7.8Hz, 1H), 7.90 (dd, J = 8.5, 1.6Hz, 1H), 7.79 - 7.74 (m, 2H), 7.61 (dd, J = 8.0, 1.4Hz, 1H), 7.47 (t, J = 7.9Hz, 1H), 6.59 - 6.55 (m, 1H), 5.85 (s, 2H), 3.68 (s, 8H); LCMS (ESI) m / z: 454.8 [M] + . Synthesis of 4-{4-[(oxan-4-yl)methoxy]-7-(pyridin-3-yl)-5H,6H,7H-pyrrolo[2,3-d]pyrimidin-2-yl}morpholine (Compound 151)

[0490] Compound 151 can be synthesized according to procedures known to those skilled in the art. [Table 21]

[0491] Example 2 PIKfyve inhibitory activity

[0492] PIKfyve Biochemical Assay. Biochemical PIKFyve inhibition assays were performed by Carna Biosciences according to a proprietary methodology based on the Promega ADP-Glo™ Kinase Assay. Full-length human PIKFYVE [1-2098 (terminal) amino acids of the protein with the sequence set forth in NCBI Reference SEQ ID NO: NP_055855.2, as well as S696N, L932S, Q995L, T998S, S1033A and Q1183K] was expressed as an N-terminal GST-fusion protein (265 kDa) using a baculovirus expression system. GST-PIKFYVE was purified by using glutathione sepharose chromatography and used in the ADP-Glo™ Kinase Assay (Promega). Reactions were set up by adding test compound solution, substrate solution, ATP solution and kinase solution at 4× final concentration, respectively. Reactions were prepared in assay buffer (50 mM MOPS, 1 mM DTT, pH 7.2), mixed and incubated at room temperature for 1 hour in a black 384-well polystyrene plate. ADP-Glo™ reagent was then added for 40 minutes, followed by kinase detection reagent for another 40 minutes. Kinase activity was assessed by detecting relative light emission in a luminescence plate reader. Samples were run in duplicate from 10 μM to 3 nM. Data was analyzed by setting control wells (+PIKfyve, no compound) to 0% inhibition and background (no PIKfyve) readings to 100% inhibition, then calculating the % inhibition for each test solution. IC50 values ​​were calculated from the concentration vs. % inhibition curves by fitting to a 4-parameter logistic curve.

[0493] NanoBRET™ TE Intracellular Kinase Assay, K-8 (Promega) cell-based assay. Intracellular inhibition of PIKfyve was assayed using Promega's NanoBRET™ TE Intracellular Kinase Assay, K-8, following the manufacturer's instructions. A dilution series of test compounds was added to HEK293 cells transfected with PIKFYVE-NanoLuc® fusion vector (Promega) containing full-length PIKfyve for a minimum of 20 hours in a 96-well plate following the manufacturer's specifications for 2 hours. Kinase activity was detected by adding NanoBRET™ tracer reagent, a proprietary PIKfyve inhibitor conjugated to a fluorescent probe (BRET, bioluminescence resonance energy transfer). Test compounds were tested at concentrations of 10, 3, 1, 0.3, 0.1, 0.03, 0.01, 0.003 μM. BRET signal was measured by GloMax Discover Multimode microplate reader (Promega) using 0.3 seconds / well integration time, 450BP donor filter and 600LP acceptor filter.Active test compounds that bind to PIKfyve and displace the above tracer reduced BRET signal.Then, IC50 value was calculated by fitting data to normalized BRET ratio.

[0494] The results of the PIKfyve inhibition assay are summarized in the table below. [Table 22-1] [Table 22-2] [Table 22-3] [Table 22-4] [Table 22-5]

[0495] Example 3 Viability assay to assess TDP-43 toxicity in FAB1 TDP-43 and PIKfyve TDP-43 yeast cells.

[0496] Generation of a TDP-43 yeast model expressing human PIKFYVE. Human PIKFYVE ("entry clone") was cloned into pAG416GPDccdB ("destination vector") following standard Gateway cloning protocols (Invitrogen, Life Technologies). The resulting pAG416GPD-PIKFYVE plasmid was amplified in E. coli and plasmid identity was confirmed by restriction digestion and Sanger sequencing. A lithium acetate / polyethylene glycol-based transformation was used to introduce the above PIKFYVE plasmid into a BY4741 yeast strain auxotrophic for the ura3 gene and express two transcription factors (MATa, snq2::KlLeu2; pdr3::Klura3; pdr1::NATMX; fab1::G418) that regulate the major xenobiotic efflux pump and the yeast orthologue of PIKFYVE. R , his3;leu2;ura3;met15;LYS2+) (Figure 2). Transformed yeast were plated on solid agar plates containing complete synthetic medium lacking uracil (CSM-ura) containing 2% glucose. Individual colonies harboring control or PIKFYVE TDP-43 plasmids were picked. A plasmid containing wild-type TDP-43 under the transcriptional control of the GAL1 promoter and a hygromycin resistance gene as a selectable marker was cloned into fab1::G418. R pAG416GPD-PIKFYVE was transformed into the yeast strain (Figure 1). After overnight recovery in antibiotic-free medium, transformed yeast were plated onto CSM-ura containing 2% glucose and 200 μg / mL G418. Multiple independent isolates were further evaluated for cytotoxicity and TDP-43 expression levels.

[0497] Viability assay. A propidium iodide viability assay was used to evaluate the toxicity of a control yeast strain containing the wild-type yeast FAB1 gene and TDP-43 ("FAB1 TDP-43" harboring an empty pAG416 plasmid), and the "PIKFYVE TDP-43" yeast strain. Both yeast strains were transferred from solid CSM-ura / 2% glucose agar plates to 3 mL of liquid CSM-ura / 2% glucose medium with aeration for 6-8 h at 30 °C. The yeast cultures were then incubated in 3 mL of CSM-ura / 2% raffinose at 4°C for 2 h at an optical density at a wavelength of 600 nm (OD 600 ) to 0.005, and incubated overnight at 30°C with aeration until the OD was 0.3-0.8. 600 Log-phase overnight cultures were grown at OD in CSM-ura containing either 2% raffinose or galactose. 600 Compounds were diluted to an affinity of 0.005 and 150 μL was dispensed into each well of a flat-bottom 96-well plate. Compounds formulated in 100% dimethyl sulfoxide (DMSO) were serially diluted in DMSO and 1.5 μL of diluted compound was transferred to the 96-well plate using a multichannel pipette. Wells containing DMSO alone were also evaluated as a control for compound effect. Test concentrations ranged from 15 μM to 0.11 μM. Cultures were immediately mixed to ensure compound distribution and the covered plates were incubated at 30°C in a stationary humidified incubator for 24 hours.

[0498] Once incubation was complete, cultures were assayed for viability using propidium iodide (PI) to stain dead / dying cells. A working solution of PI was made and for each plate, 1 μL of 10 mM PI was added to 10 mL of CSM-ura (raffinose or galactose). The final PI solution (50 μL / well) was dispensed into each well of a new round-bottom 96-well plate. The overnight 96-well assay plate was then mixed using a multichannel pipette and 50 μL was transferred to the PI-containing plate. This plate was then incubated for 30 minutes at 30° C. in the dark. A benchtop flow cytometer (Miltenyi MACSquant) was then used to assess red fluorescence (B2 channel), forward scatter, and side scatter using the following settings: gentle mixing, high flow rate, fast reading, 10,000 events. The intensity histograms were then gated for "PI positive" or "PI negative" using raffinose and galactose cultures treated with DMSO as controls. The DMSO control versus raffinose or galactose containing cultures was used to determine the window of increased cell death, and this difference was set to 100. All compounds were gated similarly and then compared to this maximum window to establish the percent decrease in PI positive cells. IC50 values ​​were then calculated for compounds that demonstrated a concentration-dependent increase in viability by fitting a logistic regression curve.

[0499] Upon induction of TDP-43 in both strains, there was a significant increase in nonviable cells (right-most population) in both FAB1 TDP-43 and PIKFYVE TDP-43 cells, with a more pronounced effect in PIKFYVE TDP-43 (Fig. 3 and 4).

[0500] In the PIKfyve TDP-43 model, PIKfyve inhibition suppresses toxicity. Biochemical PIKFyve inhibition assays were performed by Carna Biosciences according to a proprietary methodology based on the Promega ADP-Glo™ kinase assay. Full-length human PIKFYVE [1-2098 (terminal) amino acids of accession number NP_055855.2, as well as S696N, L932S, Q995L, T998S, S1033A and Q1183K] was expressed as an N-terminal GST-fusion protein (265 kDa) using a baculovirus expression system. GST-PIKFYVE was purified by using glutathione sepharose chromatography and used in the ADP-Glo™ kinase assay (Promega). Reactions were set up by adding test compound solution, substrate solution, ATP solution and kinase solution at 4× final concentration, respectively. Reactions were prepared in assay buffer (50 mM MOPS, 1 mM DTT, pH 7.2), mixed and incubated at room temperature for 1 hour in a black 384-well polystyrene plate. ADP-Glo™ reagent was then added for 40 minutes, followed by kinase detection reagent for an additional 40 minutes. Kinase activity was assessed by detecting relative luminescence in a luminescence plate reader. Samples were run in duplicate at 10 uM to 3 nM. Data was analyzed by setting control wells (+PIKfyve, no compound) to 0% inhibition and background (no PIKfyve) readings to 100% inhibition, then calculating the % inhibition for each test solution. IC50 values ​​were calculated from the concentration vs. % inhibition curves by fitting to a 4-parameter logistic curve.

[0501] Activity of APY0201, a known PIKfyve inhibitor, in FAB1 TDP-43 (Figure 5) and PIKFYVE TDP-43 (Figure 6). In FAB1 TDP-43, viable cells did not increase across the range of compound concentrations (only 0.23 μM shown), as evidenced by no reduction in the right-most population of propidium iodide positive cells. In the PIKFYVE TDP-43 model, 0.23 μM reduced the population of propidium iodide positive dead cells, indicating that PIKFYVE inhibition ameliorated TDP-43 toxicity. Concentrations ranging from 0.5 mM to less than 100 nM resulted in improved viability. [ka]

[0502] A panel of compounds was tested in the biochemical PIKFYVE assay (ADP-Glo™ using full-length PIKfyve) and IC50 was determined (nM) (see table below). The same compounds were also tested in both FAB1 and PIKFYVE TDP-43 yeast models. Their activity is reported here as "active" or "inactive". Compounds with potency at low nanomolar concentrations in the biochemical assay were active in the PIKFYVE TDP-43 yeast model. Compounds with less potency or inactive in the biochemical assay were inactive in the PIKFYVE TDP-43 model. Compounds that were inactive in the biochemical or PIKFYVE TDP-43 assay were plotted at the highest concentration tested in that assay. [Table 23-1] [Table 23-2]

[0503] Biochemical and Potency Assays. A larger series of PIKfyve inhibitors were evaluated in both the PIKfyve kinase domain binding assay (nanobret) and the PIKFYVE TDP-43 yeast strain. IC50 values ​​(μM) were plotted. Data points were formatted based on binned potency from the nanobret assay as indicated in the legend (Figure 7). Below is a table of compounds and their biochemical and PIKFYVE TDP-43 IC50 values ​​plotted in Figure 7. [Table 24-1] [Table 24-2] [Table 24-3] [Table 24-4] [Table 24-5]

[0504] Other embodiments

[0505] Various modifications and variations of the described invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are obvious to those skilled in the art are intended to be within the scope of the invention.

[0506] Other embodiments are within the claims.

Claims

1. Compounds of formula (I): 【Chemistry 143】 or a pharmaceutically acceptable salt thereof, During the ceremony, 【Chemistry 144】 is a single bond, and X １ is (C(R Ａ ) ２ ) ｍ or -OC(R Ａ ) ２ -R Ｘ and X ２ is C(R Ａ ) ２ or CO, or 【Chemistry 145】 is a double bond, and X １ and X ２ are each independently, CR Ａ or N, where R Ｘ is X ２ is a bond to R １ Is -(L) ｎ -R Ｂ halo, cyano, hydrogen, optionally substituted C １～６ Alkoxy, optionally substituted C containing at least one ring oxygen １～９ Heterocyclyl, optionally substituted C １ ~C ６ alkyl, optionally substituted piperazin-1-yl, optionally substituted pyrrolidin-3-yl, pyrimidinyl (wherein the pyrimidinyl is cyclopropyl or optionally substituted C ６ ~C １０ aryl), optionally substituted pyridazinyl, optionally substituted oxazolyl, pyrid-2-on-1-yl, optionally substituted isoindolinyl, unsubstituted pyridin-4-yl, unsubstituted pyridin-2-yl, optionally substituted furan-3-yl, unsubstituted pyridin-3-yl or optionally substituted pyrazol-1-yl; R ２ is an optionally substituted C １ ~C ６ Alkyl, optionally substituted C ６ ~C １０ Aryl, optionally substituted piperidin-4-yl, optionally substituted tetrahydropyran-4-yl, optionally substituted pyrimidin-5-yl, optionally substituted pyrimidin-4-yl, optionally substituted pyridin-3-yl, optionally substituted pyridazin-4-yl, optionally substituted pyrazol-1-yl, optionally substituted pyrazol-4-yl, optionally substituted pyrazol-3-yl, optionally substituted pyridin-2-yl, optionally substituted triazolyl, optionally substituted benzodioxol-2-yl, optionally substituted benzodioxan-2-yl, optionally substituted C ６ ~C １０ Aryl C １ ~C １０ alkyl or optionally substituted acyl; R ３ is a group of the following structure: 【Chemistry 146】 and R Ａ each independently represents H, optionally substituted C １～６ Alkyl or optionally substituted C ６ ~C １０ aryl or two R Ａ together with the atoms to which they are attached form an oxo, where two R Ａ When they combine with the atom to which they are attached to form an oxo, 【Chemistry 147】 is a single bond, R Ｂ is an optionally substituted C ６～１０ Aryl, optionally substituted C １ ~C ９ Heteroaryl, optionally substituted C ３～８ Cycloalkyl, —N═CH—R Ｄ or optionally substituted C １ ~C ９ Heterocyclyl, optionally substituted C ２ ~C ９ Heteroaryl C １ ~C ６ Alkyl, optionally substituted C ２ ~C ９ Heterocyclyl C １ ~C ６ is alkyl, R Ｃ is H or optionally substituted C １ ~C ６ is alkyl, R Ｄ is an optionally substituted C ６ ~C １０ is aryl, or Each L is independently an optionally substituted C １～６ Alkylene, optionally substituted C １ ~C ６ Heteroalkylene, optionally substituted C ３ ~C ８ Cycloalkylene, optionally substituted C ２ ~C ６ Alkynylene, O or NR Ｃ and n is 1, 2 or 3; m is 0, 1 or 2; The compound, or a pharmaceutically acceptable salt thereof.

2. The compound has formula (1a), (1a'), (1b), (1c), (1d), or (1e): 【Chemistry 149】 [Chemical 150] 【Chemistry 151】 【Chemistry 152】 【Chemistry 153】 or 【Chemistry 154】 2. The compound of claim 1, which is a compound of formula (I): or a pharmaceutically acceptable salt thereof.

3. R １ is optionally substituted C １ ~C ６ The compound of claim 1 or 2, which is alkyl or optionally substituted piperazin-1-yl.

4. R ２ is optionally substituted C ６ ~C １０ Aryl C １ ~C ６ The compound of claim 1, wherein the alkyl is an optionally substituted C 1 -C 6 alkyl, or an optionally substituted C 6 -C 10 aryl.

5. R １ but, 【Chemistry 155】 【Chemistry 156】 2. The compound of claim 1, which is methyl or methoxy.

6. R ２ but, 【Chemistry 158】 2. The compound of claim 1, wherein:

7. The compound has the structure: 【Chemistry 161】 or a pharmaceutically acceptable salt thereof (In the formula, R ２ is optionally substituted pyrimidin-3-yl or optionally substituted pyrimidin-4-yl; R ４ is hydrogen or optionally substituted C ６ ~C １０ aryl) or The compound has the structure: 【Chemistry 162】 or a pharmaceutically acceptable salt thereof wherein R 5 is hydrogen or optionally substituted C 6 -C 10 aryl; R 2 is optionally substituted triazolyl, optionally substituted pyrazol-4-yl, optionally substituted pyrazol-3-yl, optionally substituted pyrimidin-4-yl or optionally substituted C 6 -C 10 arylC 1 -C 6 alkyl. or The compound has the structure: 【Chemistry 165】 or a pharmaceutically acceptable salt thereof wherein R 2 is optionally substituted pyridin-3-yl. or The compound has the structure: 【Chemistry 166】 or a pharmaceutically acceptable salt thereof wherein R 2 is optionally substituted C 6 -C 10 aryl; or optionally substituted pyridin-4-yl. or The compound has the structure: 【Chemistry 167】 or a pharmaceutically acceptable salt thereof wherein R 2 is optionally substituted pyridin-3-yl. or The compound has the structure: 【Chemical 168】 or a pharmaceutically acceptable salt thereof wherein L is an optionally substituted C 3 -C 8 cycloalkylene or C 2 -C 6 alkynylene; R B is an optionally substituted C 6 -C 10 aryl. or The compound has the structure: 【Chemistry 171】 or a pharmaceutically acceptable salt thereof (wherein L-R B is —NHN═CHR D ; R 2 is optionally substituted pyridin-3-yl; R D is an optionally substituted C 6 -C 10 aryl. or The compound has the structure: 【Chemistry 173】 or a pharmaceutically acceptable salt thereof wherein R 2 is optionally substituted pyridin-3-yl; R 6 is an optionally substituted C 6 -C 10 aryl.

2. The compound of claim 1 having the formula:

8. The compound of claim 7, wherein R 4 is hydrogen or optionally substituted C 6 -C 10 aryl.

9. The compound of claim 7 or 8, wherein R 2 is optionally substituted pyridin-3-yl or pyridin-4-yl, and R 5 is hydrogen or phenyl.

10. structure: 【Chemistry 174】 or a pharmaceutically acceptable salt thereof, In the formula, X ２ is N or CH, R Ａ is an optionally substituted C ２ ~C ９ heteroaryl; or optionally substituted C ６ ~C １０ is aryl, R １ is an optionally substituted C ２ ~C ９ Heteroaryl or -O-R ７ and R ２ is hydrogen or optionally substituted C １ ~C ６ is alkyl, R ７ is an optionally substituted C ２ ~C ９ Heteroaryl C １ ~C ６ is alkyl, The compound, or a pharmaceutically acceptable salt thereof.

11. The compound has the structure: 【Chemistry 175】 or a pharmaceutically acceptable salt thereof, or The compound has the structure: 【Chemistry 176】 or a pharmaceutically acceptable salt thereof, or The compound has the structure: 【Chemistry 177】 or a pharmaceutically acceptable salt thereof.

12. R Ａ is optionally substituted pyridin-2-yl, optionally substituted pyrimidin-4-yl, optionally substituted pyrimidin-2-yl, optionally substituted pyrazol-4-yl, optionally substituted pyridin-3-yl, optionally substituted pyrazol-4-yl, optionally substituted 7-aza-5,6,7,8-tetrahydroindolizin-1-yl, optionally substituted pyridazin-3-yl or optionally substituted pyridin-4-yl.

13. A compound of the following structure: Table 27-1 Table 27-2 Table 27-3 Table 27-4 Table 27-5 Table 27-6 Table 27-7 Table 27-8 Table 27-9 Table 27-10 Table 27-11 Table 27-12 Table 27-13 Table 27-14 or a pharmaceutically acceptable salt thereof.

14. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

15. A pharmaceutical composition for use in the treatment of neurological disorders, comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof.

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