Triazole compound as well as preparation method and medical application thereof

Abstract

Compounds of formula (I) useful as WRN inhibitors, processes for their preparation, pharmaceutical compositions comprising these compounds, and pharmaceutical uses for the treatment of WRN-mediated diseases or disorders are disclosed. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicine and relates to triazole compounds, methods for their preparation, pharmaceutical compositions containing these compounds, and their medical uses. Background Art

[0002] Cells within any living organism possess multiple DNA repair mechanisms to maintain genomic stability and integrity, which are essential for survival. DNA mismatch repair (MMR) is a highly conserved pathway that plays a crucial role in maintaining genomic stability by correcting errors that occur during DNA replication, recombination, and repair. (Pecina-Slaus, N., Kafka, A., Salamon, I., and Bukovac, A. Mismatch Repair Pathway, Genome Stability and Cancer [Mismatch Repair Pathway, Genome Stability and Cancer]. Front Mol. Biosci. [Frontiers in Molecular Biosciences] 7, 122 (2020)). Defects in the MMR mechanism result in genomic hypermutation and instability, manifested as high-frequency insertions and deletions on short tandem repeat DNA sequences (microsatellites) throughout the genome, a phenomenon known as microsatellite instability (MSI). (Kim, T. M., Laird, P. W., and Park, P. J. The landscape of microsatellite instability in colorectal and endometrial cancer genomes [Overview of Microsatellite Instability in Colorectal and Endometrial Cancer Genomes]. Cell [Cell] 155, 858 - 868 (2013); Vernole, P. et al. Common fragile sites in colon cancer cell lines: role of mismatch repair, RAD51 and poly(ADP-ribose) polymerase-1 [Role of Mismatch Repair, RAD51, and Poly(ADP-Ribose) Polymerase-1 in Common Fragile Sites in Colon Cancer Cell Lines]. Mutat. Res [Mutation Research] 712, 40 - 48 (2011)). Microsatellites are generated by DNA polymerase slippage during replication or mismatch repair and are prone to misalignment and frameshift mutations, which can be recognized and corrected by the MMR process in normal cells. However, in cancer cells lacking a normal MMR mechanism, errors that occur during replication or repair accumulate, and the mutation rate in the genome increases (Lower, S. S., McGurk, M. P., Clark, A. G., et al., Satellite DNA evolution: old ideas, new approaches [Satellite DNA Evolution: Old Ideas, New Approaches]. Curr. Opin. Genet. Dev.[New Insights into Genetics and Development], 2018; 49: 70-78). High frequencies of deletions and insertions in microsatellites lead to MSI, which causes 10%-30% of ovarian, colorectal, gastric, and endometrial cancers (Aaltonen, L.A. et al. Clues to the pathogenesis of familial colorectal cancer, Science 260, 812-816 (1993); Bonneville R et al., Landscape of Microsatellite Instability Across 39 Cancer Types. JCO Precis Oncol. 1: PO.17.00073 (2017)). In contrast to MSS cancers, cancer patients with high MSI (MSI-H) show better overall prognosis and reduced metastatic potential, along with higher tumor mutation burden and immunogenicity (Kang, S. et al. The significance of microsatellite instability in colorectal cancer after controlling for clinicopathological factors. Med. (Baltim.) 97, e0019 (2018)), and MSI-H tumors tend to be resistant to immunotherapy and chemotherapy (Le, D.T. et al. Phase II Open-Label Study of Pembrolizumab in Treatment-Refractory, Microsatellite Instability-High / Mismatch Repair-Deficient Metastatic Colorectal Cancer: KEYNOTE-164. J. Clin. Oncol. 38, 11-19 (2020); Overman, M.J.et al. Nivolumab in patients with metastatic DNA mismatch repair-deficient or microsatellite instability-high colorectal cancer (CheckMate 142): an open-label, multicentre, phase 2 study [Nivolumab in patients with metastatic DNA mismatch repair-deficient or microsatellite instability-high colorectal cancer (CheckMate 142): an open-label, multicentre, phase 2 study]. Lancet Oncol. [Lancet-Oncology] 18, 1182-1191 (2017); Fuca, G. et al. Ascites and resistance to immune checkpoint inhibition in dMMR / MSI-H metastatic colorectal and gastric cancers [Ascites and resistance to immune checkpoint inhibition in dMMR / MSI-H metastatic colorectal and gastric cancers]. J. Immunother. Cancer [Journal of Cancer Immunotherapy] 10, 4001 (2022)).

[0003] Recently, multiple independent large-scale functional genomics screens using over 300 human cancer cell lines have identified the Werner syndrome RecQ helicase (WRN) as selectively required for the survival of MSI-H cells (Behan, F.M. et al. Prioritization of cancer therapeutic targets using CRISPR-Cas9 screens. Nature 568, 511-516 (2019); McDonald E.R. et al., Project DRIVE: A Compendium of Cancer Dependencies and Synthetic Lethal Relationships Uncovered by Large-Scale, Deep RNAi Screening. Cell 170(3):577-592 (2017); Chan, E.M. et al. WRN helicase is a synthetic lethal target in microsatellite unstable cancers. Nature 568, 551-556 (2019)).WRN, one of the five human RecQ-like helicases, is a multifunctional enzyme with helicase and exonuclease activities and plays an important role in multiple pathways of DNA repair and maintenance of genome integrity, including DNA replication, transcription, DNA repair, and telomere maintenance (Bohr, VARising from the RecQ-age: the role of human RecQ helicases in genome maintenance. Trends Biochem Sci. 33, 609-620 (2008); Singh, DK, Ahn, B. and Bohr, V.A. Roles of RECQ helicases in recombination based DNA repair, genomic stability and aging. Biogerontology 10, 235-252 (2009); Rossi, ML, Ghosh, AK and Bohr, V.A. Roles of RECQ helicases in recombination based DNA repair, genomic stability and aging. Biogerontology 10, 235-252 (2009); Rossi, ML, Ghosh, AK and Bohr, V.A. Roles of RECQ helicases in recombination based DNA repair, genomic stability and aging. Biogerontology 10, 235-252 (2009); Rossi, ML, Ghosh, AK and Bohr, V.A. Roles of RECQ helicases in recombination based DNA repair, genomic stability and aging. of Werner syndrome protein inprotection of genome integrity. DNA Repair (Amst.) 9, 331-344 (2010).

[0004] Loss of WRN results in various defects in cellular and genomic structure, including cell cycle arrest, DNA breaks, mitotic defects, chromothripsis, and apoptosis in MSI but not MSS cell lines. (Behan, F.M. et al. Prioritization of cancer therapeutic targets using CRISPR-Cas9 screens. Nature 568, 511-516 (2019); McDonald E.R. et al., Project DRIVE: A Compendium of Cancer Dependencies and Synthetic Lethal Relationships Uncovered by Large-Scale, Deep RNAi Screening. Cell 170(3):577-592 (2017); Chan, E.M. et al. WRN helicase is a synthetic lethal target in microsatellite unstable cancers. Nature 568, 551-556 (2019)). Additionally, loss-of-function mutation and deletion studies of WRN further pinpoint that the helicase function of WRN is indispensable for the survival of MSI cells (Lieb, S. et al. Werner syndrome helicase is a selective vulnerability of microsatellite instability-high tumor cells. Elife 8, e43333 (2019)). These results clearly suggest that targeting WRN (especially the helicase domain) can be a promising strategy for treating MSI-high cancers. SUMMARY OF THE INVENTION

[0005] In one aspect, the present invention provides a compound of formula (I)-(XI), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, including its tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereomer, or mixture:

[0006]

[0007] or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein

[0008] Ring A and Ring B are each independently selected from aryl or heteroaryl;

[0009] Y1 is CH, NH, N, O, or S;

[0010] Y2 is CH, NH, N, O, or S;

[0011] Y3 is CH, NH, N, O, or S;

[0012] X1 and X2 are each independently CH or N;

[0013] M1, M2, M5, M6 are each independently CH, NH, N, O, or S;

[0014] M3 and M4 are each independently -NR a , -CR a R b ;

[0015] R a and R b are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, and hydroxyalkyl;

[0016] Y2 is CH, NH, N, O, or S;

[0017] Y3 is CH, NH, N, O, or S;

[0018] is a single bond or a double bond, provided that are not both double bonds;

[0019] R1, R3, and R4 are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, and hydroxyalkyl;

[0020] Two R2s together with the atoms to which they are attached form a C3-C 10 cycloalkyl or a C3-C 10 heterocyclic group,

[0021] R5 and R6 are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, and hydroxyalkyl;

[0022] or R5 and R6 together with the atoms to which they are attached form a C3-C 10 cycloalkyl, C3-C 10 heterocycloalkyl, C6-C 14 aryl or a C5-C containing 1 to 4 heteroatoms selected from the group consisting of O, S and N 12 heteroaryl;

[0023] R7 is selected from the group consisting of: hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0024] or two R7 together with the atoms to which they are attached form a C3-C 10 cycloalkyl or C3-C 10 heterocycloalkyl,

[0025] L1 is a bond, O, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl, -C3-C 10 cycloalkyl-CONH-, -C1-C6 alkyl-C3-C 10 cycloalkyl-CONH, -C1-C6 haloalkyl-NH-, -C3-C 10 -heterocyclo-NH-alkyl, -C3-C 10 cycloalkyl-NH-, -C1-C6 alkyl-C(NH)-NH, -C6-C 10 heteroaryl-C1-C6 alkyl, -S(O)2-NH- and -C1-C6 alkyl-S(O)2-NH-;

[0026] L2 is a bond, O, -C(O), -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl and -CON(R c )-;

[0027] R c is selected from the group consisting of: hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0028] Ring A is C6-C 14 aryl or a C5-C containing 1 to 4 heteroatoms selected from the group consisting of O, S and N 12 heteroaryl;

[0029] m is 0, 1, 2, 3 or 4;

[0030] p is 0, 1, 2 or 3;

[0031] q is 0, 1, 2 or 3;

[0032] r is 0, 1, 2 or 3;

[0033] s is 2 or 3;

[0034] t is 0, 1, 2 or 3;

[0035] w is 0, 1, 2 or 3; and

[0036] n is 0, 1, 2 or 3.

[0037] In one embodiment, the compound has the formula (XII)-(XIII), (XIX)-(XXI)

[0038] R8 is C3-C 12 fused cycloalkyl, C 2- C 10 fused heterocyclic group, wherein the fused cycloalkyl and fused heterocyclic group are each optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, oxo;

[0039] R9 is a bridged C3-C 10 cycloalkyl or a bridged C3-C 10 heterocyclic group, wherein the bridged cycloalkyl and bridged heterocyclic group are each optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxy, C 1-5 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, oxo, C 1-6 haloalkyl, C 3-5 cycloalkyl;

[0040] L6 is a bridged C3-C 10 cycloalkyl or a bridged C2-C 10 heterocyclic group, wherein the bridged cycloalkyl and bridged heterocyclic group are each optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, oxo, C 1-6 haloalkyl, C 3-5 cycloalkyl;

[0041] Y1, Y2, Y3, M1, M2, L2, X1, X2, R1, R3, R4, m, p, q, r, t are as defined above.

[0042] In one embodiment, R1, R3 and R4 are each independently selected from the group consisting of: hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 hydroxyalkyl and C2-C6 alkenyl;

[0043] In a preferred embodiment, R1, R3 and R4 are each independently selected from the group consisting of: hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, -CF3.

[0044] In one embodiment, the compound has the formula (XXII) or (XXIII),

[0045]

[0046] Each X is independently selected from F, Cl, Br, I;

[0047] Y1, Y2, Y3, M1, M2, L2, X1, X2, R1, R3, R7, m, n, p, r, t, w are as defined above.

[0048] In one embodiment, two R2 together with the atoms to which they are attached form a bridged C3-C 10 cycloalkyl or a bridged C3-C 10 heterocyclic group;

[0049] In a preferred embodiment, represents

[0050] In one embodiment, R5 and R6 are each independently selected from the group consisting of: hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl;

[0051] or R5 and R6 together with the atoms to which they are attached form

[0052] In one embodiment, R7 is selected from the group consisting of: hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl;

[0053] or two R7 together with the atoms to which they are attached form a bridging C3-C 10 cycloalkyl or a bridging C3-C 10 heterocyclic group, preferably denotes

[0054] In one embodiment, R a and R b are each independently selected from the group consisting of: hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl;

[0055] In one embodiment, L1 is a bond, O, C1-C3 alkyl, C2-C3 alkenyl, -C1-C3 alkyl-NH-, -C2-C3 alkenyl-NH-, C2-C3 alkynyl, C1-C3 alkoxy, C1-C3 alkylthio, C1-C3 haloalkyl, -C3-C6 cycloalkyl-CONH-, -C1-C3 alkyl-C3-C6 cycloalkyl-CONH, -C1-C3 haloalkyl-NH-, -C3-C6 -heterocyclic-NH-alkyl, -C3-C6 cycloalkyl-NH-, -C1-C6 alkyl-C(NH)-NH, -C6-C 10 heteroaryl-C1-C6 alkyl, -S(O)2-NH- and -C1-C6 alkyl-S(O)2-NH-;

[0056] In one embodiment, L2 is a bond, O, -C(O), -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C3 alkyl-NH-, -C2-C3 alkenyl-NH-, C2-C3 alkynyl, C1-C3 alkoxy, C1-C3 alkylthio, C1-C3 haloalkyl and -CON(R c );

[0057] R c is selected from the group consisting of: hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl;

[0058] In one embodiment, ring A is

[0059] In one embodiment, ring B is selected from the group consisting of: phenyl, pyridyl, pyrrolyl or pyrimidinyl.

[0060] In one embodiment, is

[0061] In one embodiment, R8 is selected from the group consisting of:

[0062] In one embodiment, R9 is R10 is C 1-5 alkyl, C 3-5 cycloalkyl.

[0063] In one embodiment, L6 is or L6-L2 is

[0064] In one embodiment, in formula (XXII), X connected to the phenyl group is Cl, and / or CH n X 3-n is CF3.

[0065] In one embodiment, in formula (XXIII), CH n X 3-n is CF3.

[0066] The present invention also provides a pharmaceutical composition, which comprises a therapeutically effective amount of a compound of any one of formulas (I)-(X), or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, together with one or more pharmaceutically acceptable carriers, diluents or excipients.

[0067] In another aspect, the present invention relates to a method for treating a WRN-mediated disease, which comprises administering to a subject in need thereof an effective amount of a compound of any one of formulas (I)-(X), or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a pharmaceutical composition comprising the same.

[0068] In another aspect, the present invention relates to a method for treating cancer, which comprises administering to a subject in need thereof an effective amount of a compound of any one of formulas (I)-(X), or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a pharmaceutical composition comprising the same.

[0069] In a preferred embodiment, the cancer is selected from the group consisting of: microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR), selected from colorectal cancer, gastric cancer, prostate cancer, endometrial cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, renal cancer and ovarian cancer. Detailed Description

[0070] The definitions of the terms used in this application are given below. Any term not defined herein shall have the ordinary meaning as understood by a person skilled in the art.

[0071] "Alkyl" means including C1-C 20Saturated aliphatic hydrocarbon groups of straight-chain and branched-chain groups. Preferably, the alkyl group is an alkyl group having 1 to 12, sometimes preferably 1 to 6, and sometimes more preferably 1 to 4 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and their branched isomers. More preferably, the alkyl group is a lower alkyl group having 1 to 6 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. The alkyl group can be substituted or unsubstituted. When substituted, one or more substituents can be substituted at any available attachment point, and preferably one or more substituents are one or more substituents independently selected from the group consisting of alkyl, halogen, alkoxy, alkenyl, alkynyl, alkylsulfonic acid group, alkylamino, thiol, hydroxy, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocyclic group, cycloalkylthio, heterocycloalkylthio, and oxo group.

[0072] "Alkenyl" means an alkyl group having at least two carbon atoms and at least one carbon-carbon double bond as defined above, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc., preferably C 2-20 alkenyl, more preferably C 2-12 alkenyl, and most preferably C 2-6 alkenyl. The alkenyl group may be substituted or unsubstituted. When substituted, one or more substituents are preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three groups independently selected from the group consisting of: alkyl, halogen, alkoxy, alkenyl, alkynyl, alkylsulfonyl, alkylamino, mercapto, hydroxy, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocyclic group, cycloalkylthio, heterocycloalkylthio and oxo group.

[0073] "Alkynyl" means an alkyl group having at least two carbon atoms and at least one carbon-carbon triple bond as defined above, such as ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, etc., preferably C 2-20 alkynyl, more preferably C 2-12 alkynyl, and most preferably C 2-6 alkynyl. The alkynyl group may be substituted or unsubstituted. When substituted, one or more substituents are preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three groups independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio.

[0074] "Alkylene" refers to a saturated straight-chain or branched aliphatic hydrocarbon group having two residues derived by removing two hydrogen atoms from the same carbon atom or two different carbon atoms of the parent alkane. A straight-chain or branched group containing 1 to 20 carbon atoms preferably has 1 to 12 carbon atoms, more preferably 1 to 6 carbon atoms. Non-limiting examples of alkylene include, but are not limited to, methylene (-CH2-), 1,1-ethanediyl (-CH(CH3)-), 1,2-ethanediyl (-CH2CH2-), 1,1-propanediyl (-CH(CH2CH3)-), 1,2-propanediyl (-CH2CH(CH3)-), 1,3-propanediyl (-CH2CH2CH2-), 1,4-butanediyl (-CH2CH2CH2CH2-), etc. Alkylene may be substituted or unsubstituted. When substituted, one or more substituents are preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio.

[0075] "Alkenylene" refers to an alkylene having at least two carbon atoms and at least one carbon-carbon double bond as defined above, preferably C 2-20 alkenylene, more preferably C 2-12 alkenylene, and most preferably C 2-6 alkenylene. Non-limiting examples of alkenylene include, but are not limited to, -CH=CH-, -CH=CHCH2-, -CH=CHCH2CH2-, -CH2CH=CHCH2-, etc. Alkenylene may be substituted or unsubstituted. When substituted, one or more substituents are preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio.

[0076] "Alkynylene" refers to an alkynyl having at least two carbon atoms and at least one carbon-carbon triple bond as defined above, preferably C 2-20 alkynylene, more preferably C 2-12 alkynylene, and most preferably C 2-6Alkynylene. Non-limiting examples of alkynylene include, but are not limited to, -CH≡CH-, -CH≡CHCH2-, -CH≡CHCH2CH2-, -CH2CH≡CHCH2-, etc. The alkynylene group can be substituted or unsubstituted. When substituted, one or more substituents are preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonic acid group, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio.

[0077] "Cycloalkyl" refers to a saturated and / or partially unsaturated monocyclic or polycyclic hydrocarbon group having 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 8 carbon atoms or 3 to 6 carbon atoms. Representative examples of monocyclic cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc. Polycyclic cycloalkyl includes cycloalkyl having spiro rings, fused rings, or bridged rings.

[0078] "Spirocycloalkyl" refers to a 5- to 20-membered polycyclic group in which the rings are joined by a common carbon atom (referred to as a spiro atom), where one or more of the rings may contain one or more double bonds, but no ring has a completely conjugated π-electron system. Preferably, the spirocycloalkyl is 6 to 14 members and more preferably 7 to 10 members. Depending on the number of shared spiro atoms, the spirocycloalkyl is classified as monospirocycloalkyl, bisspirocycloalkyl, or polyspirocycloalkyl, and preferably refers to monospirocycloalkyl or bisspirocycloalkyl, more preferably 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered monospirocycloalkyl. Representative examples of spirocycloalkyl include, but are not limited to, the following substituents:

[0079]

[0080] "Fused cycloalkyl" refers to a 5- to 20-membered polycyclic hydrocarbon group in which each ring in the system shares a pair of adjacent carbon atoms with another ring, where one or more of the rings may contain one or more double bonds, but no ring has a completely conjugated π-electron system. Preferably, the fused cycloalkyl is 6 to 14 members, more preferably 7 to 10 members. Depending on the number of member rings, the fused cycloalkyl is classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused cycloalkyl, and preferably refers to bicyclic or tricyclic fused cycloalkyl, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused cycloalkyl. Representative examples of fused cycloalkyl include, but are not limited to, the following substituents:

[0081]

[0082] "Bridged cycloalkyl" refers to a 5- to 20-membered polycyclic hydrocarbon group in which every two rings in the system share two non-adjacent carbon atoms. These rings may have one or more double bonds, but do not have a fully conjugated π-electron system. Preferably, the bridged cycloalkyl is 6- to 14-membered and more preferably 7- to 10-membered. Depending on the number of member rings, bridged cycloalkyls are classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyls, and preferably refer to bicyclic, tricyclic or tetracyclic bridged cycloalkyls, more preferably bicyclic or tricyclic bridged cycloalkyls. Representative examples of bridged cycloalkyls include, but are not limited to, the following substituents:

[0083]

[0084] The cycloalkyl may be fused to a ring of an aryl, heteroaryl or heterocycloalkyl, wherein the ring bonded to the parent structure is the cycloalkyl. Representative examples include, but are not limited to, indanylacetic acid, tetralin, benzocycloheptyl, etc. The cycloalkyl is optionally substituted or unsubstituted. When substituted, one or more substituents are preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three substituents, which are independently selected from the group consisting of: alkyl, halogen, alkoxy, alkenyl, alkynyl, alkylsulfonyl, alkylamino, thiol, hydroxy, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocyclic group, cycloalkylthio, heterocycloalkylthio and oxo group.

[0085] "Heterocyclic group" refers to a 3- to 20-membered saturated and / or partially unsaturated monocyclic or polycyclic hydrocarbon group having one or more, sometimes preferably one to five, and sometimes more preferably one to three heteroatoms selected from the group consisting of N, O and S(O) m (where m is 0, 1 or 2) as ring atoms, but the ring does not include -O-O-, -O-S- or -S-S-, and the remaining ring atoms are C. Preferably, the heterocyclic group is a 3- to 12-membered heterocyclic group having 1 to 4 heteroatoms; more preferably a 3- to 10-membered heterocyclic group having 1 to 3 heteroatoms; more preferably a 4- to 8-membered heterocyclic group having 1 to 3 heteroatoms; most preferably a 5- to 6-membered heterocyclic group having 1 to 2 heteroatoms. Representative examples of monocyclic heterocyclic groups include, but are not limited to, oxetanyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, sulfonyl-morpholinyl, homopiperazinyl, etc. Polycyclic heterocyclic groups include heterocyclic groups having spiro rings, fused rings or bridged rings.

[0086] "Spiro heterocyclic group" refers to a 5- to 20-membered polycyclic heterocyclic group in which the rings are connected by a shared carbon atom (referred to as a spiro atom), wherein the rings have one or more, sometimes preferably one to five, and sometimes more preferably one to three heteroatoms selected from the group consisting of N, O and S(O) ma heteroatom selected from the group consisting of 0, 1 or 2) as a ring atom, and the remaining ring atoms are C, where one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. Preferably, the spiroheterocyclic group is 6 to 14 membered and more preferably 7 to 10 membered. Depending on the number of shared spiro atoms, the spiroheterocyclic group is classified as a monospiroheterocyclic group, a dispiroheterocyclic group or a polyspiroheterocyclic group, and preferably refers to a monospiroheterocyclic group or a dispiroheterocyclic group, more preferably a 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered or 5-membered / 6-membered monospiroheterocyclic group. Representative examples of the spiroheterocyclic group include, but are not limited to, the following substituents:

[0087]

[0088] "Fused heterocyclic group" means a 5- to 20-membered polycyclic heterocyclic group in which each ring in the system shares a pair of adjacent carbon atoms with another ring, where one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system, and where said rings have one or more, sometimes preferably one to five, sometimes more preferably one to three heteroatoms selected from the group consisting of N, O and S(O) p (where p is 0, 1 or 2) as ring atoms, and the remaining ring atoms are C. Preferably, the fused heterocyclic group is 6 to 14 membered and more preferably 7 to 10 membered. Depending on the number of member rings, the fused heterocyclic group is classified as a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably referring to a bicyclic or tricyclic fused heterocyclic group, more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic group. Representative examples of the fused heterocyclic group include, but are not limited to, the following substituents:

[0089]

[0090] "Bridged heterocyclic group" means a 5- to 14-membered polycyclic heterocycloalkyl group in which every two rings in the system share two non-adjacent atoms, the rings may have one or more double bonds, but do not have a fully conjugated π-electron system, and the rings have one or more heteroatoms selected from the group consisting of N, O and S(O) m (where m is 0, 1 or 2) as ring atoms, and the remaining ring atoms are C. Preferably, the bridged heterocyclic group is 6 to 14 membered and more preferably 7 to 10 membered. Depending on the number of member rings, the bridged heterocyclic group is classified as a bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic group, and preferably refers to a bicyclic, tricyclic or tetracyclic bridged heterocyclic group, more preferably a bicyclic or tricyclic bridged heterocyclic group. Representative examples of the bridged heterocyclic group include, but are not limited to, the following substituents:

[0091]

[0092] The ring of the heterocyclic group may be fused with the ring of an aryl, heteroaryl or cycloalkyl group, wherein the ring bonded to the parent structure is a heterocyclic group. Representative examples include, but are not limited to, the following substituents:

[0093] etc.

[0094] The heterocyclic group is optionally substituted or unsubstituted. When substituted, one or more substituents are preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three groups, which are independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonic acid group, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio.

[0095] "Aryl" refers to a 6- to 14-membered fully carbon monocyclic or polycyclic fused ring ("fused" ring system means that each ring in the system shares a pair of adjacent carbon atoms with another ring in the system) group and has a fully conjugated π electron system. Preferably, the aryl is 6 to 10 membered, such as phenyl and naphthyl, and most preferably phenyl. The aryl may be fused with the ring of a heteroaryl, heterocyclic group or cycloalkyl group, wherein the ring bonded to the parent structure is an aryl. Representative examples include, but are not limited to, the following substituents:

[0096]

[0097] The aryl may be substituted or unsubstituted. When substituted, one or more substituents are preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three substituents, which are independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonic acid group, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio.

[0098] "Heteroaryl" refers to an aryl system having 1 to 4 heteroatoms selected from the group consisting of O, S and N as ring atoms and having 5 to 14 ring atoms. Preferably, the heteroaryl is 5 to 10 membered, more preferably 5 or 6 membered, such as thiazolyl, pyrazolyl, oxazolyl, oxadiazolyl, imidazolyl, triazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl may be fused with the ring of an aryl, heterocyclic group or cycloalkyl group, wherein the ring bonded to the parent structure is a heteroaryl. Representative examples include, but are not limited to, the following substituents:

[0099]

[0100] The heteroaryl may be substituted or unsubstituted. When substituted, one or more substituents are preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three substituents, which are independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, and -NR 9 R 10 .

[0101] "Alkoxy" refers to the -O-(alkyl) and -O-(unsubstituted cycloalkyl) groups, where alkyl is as defined above. Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, etc. The alkoxy may be substituted or unsubstituted. When substituted, the substituent is preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three substituents, which are independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio.

[0102] "Bond" refers to a covalent bond, denoted by the symbol "-".

[0103] "Hydroxyalkyl" refers to an alkyl substituted with a hydroxy group, where alkyl is as defined above.

[0104] "Hydroxy" refers to the -OH group.

[0105] "Halogen" refers to a fluorine, chlorine, bromine, or iodine atom.

[0106] "Amino" refers to the -NH2 group.

[0107] "Cyano" refers to the -CN group.

[0108] "Nitro" refers to the -NO2 group.

[0109] "Oxo group" refers to the =O group.

[0110] "Carboxyl" refers to the -C(O)OH group.

[0111] "Alkoxycarbonyl" refers to the -C(O)O(alkyl) or -C(O)O(cycloalkyl) groups, where alkyl and cycloalkyl are as defined above.

[0112] "Optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and the description includes the situation where the event or circumstance may or may not occur. For example, "a heterocyclic group optionally substituted by an alkyl group" means that the alkyl group may, but need not, be present, and the description includes the cases where the heterocyclic group is substituted by an alkyl group and where the heterocyclic group is not substituted by an alkyl group.

[0113] "Substituted" means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, are independently replaced by an equal number of substituents. It goes without saying that the substituents are only present at their possible chemical positions. A person skilled in the art can determine whether substitution is possible without undue effort, either experimentally or theoretically. For example, the combination of an amino or hydroxyl group with a free hydrogen and a carbon atom with an unsaturated bond (such as an olefinic carbon atom) may be unstable.

[0114] "Pharmaceutical composition" means a mixture of one or more compounds described in the present invention or their physiologically / pharmaceutically acceptable salts or prodrugs with other chemical components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to an organism, favor the absorption of the active ingredient, and thus exert a biological activity.

[0115] "Pharmaceutically acceptable salt" means a salt of a compound of the present invention, such salts being safe and effective when used in mammals and having the corresponding biological activity.

[0116] Dosage form: For a subject weighing about 50 - 70 kg, the pharmaceutical composition or combination of the present invention may be, for example, a unit dose of about 1 - 1000 mg of one or more active ingredients.

[0117] In some embodiments, in the above-mentioned pharmaceutical composition, the amount of the compound, its tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereomer, or mixture, or its pharmaceutically acceptable salt is about 0.1% - 95% by weight of the free base; preferably about 5% - 70%, for example, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%.

[0118] In some embodiments, the above-mentioned pharmaceutical composition is formulated into tablets, capsules, liquid form, or injection form.

[0119] In some embodiments, in the above-mentioned pharmaceutical composition, the amount of the compound, its tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereomer, or mixture, or its pharmaceutically acceptable salt is about 1-1000 mg, 0.1%-95% by weight of the free base; preferably about 1-500 mg, more preferably about 1 mg, 2 mg, 3 mg, 5 mg, 10 mg, 20 mg, 40 mg, 50 mg, 60 mg, 80 mg, 100 mg, 200 mg, 300 mg, 400 mg or 500 mg.

[0120] In some embodiments, the compound, its tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereomer, or mixture, or its pharmaceutically acceptable salt can be administered by any suitable route of administration, such as oral, parenteral, buccal, sublingual, nasal, rectal, intrathecal or transdermal administration, and the pharmaceutical composition is adjusted accordingly.

[0121] In some embodiments, the compound, its tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereomer, or mixture, or its pharmaceutically acceptable salt is formulated into a solid or liquid form, such as syrup, suspension, emulsion, tablet, capsule, powder, granule or lozenge.

[0122] In the compounds of the above formula, the atoms can exhibit their natural isotope abundances, or one or more atoms can be artificially enriched with specific isotopes having the same atomic number but different atomic masses or mass numbers from those predominantly found in nature. The present disclosure as described and claimed herein is intended to include all suitable isotopic variants of the compounds of the above formula and their embodiments. For example, different isotopic forms of hydrogen (H) include protium (1H) and deuterium (2H, also denoted as D herein). Protium is the major hydrogen isotope found in nature. Enrichment of deuterium can provide certain therapeutic advantages, such as increased in vivo half-life or reduced dose requirements, or can provide compounds that can be used as standards for the characterization of biological samples. Isotope-enriched compounds can be prepared by conventional techniques well known to those skilled in the art or by methods similar to those described in the protocols and examples herein, using appropriate isotope-enriched reagents and / or intermediates, without undue experimentation.

[0123] Examples

[0124] Synthetic procedures

[0125] Synthesis of intermediates

[0126] Synthesis of ethyl 2-bromo-3-oxopentanoate 1nt 1

[0127]

[0128] To a solution of ethyl 3-oxopentanoate (15 g, 104 mmol) in dichloromethane (150 mL) was added NBS (19.4 g, 109.2 mmol), and then TsOH·H2O (3.96 g, 20.8 mmol) was added. The reaction mixture was stirred at room temperature for 2.5 h. The mixture was filtered and the filter cake was washed with dichloromethane. The combined filtrate and washings were washed with water. The aqueous layer was back-extracted with dichloromethane. The combined organic layers were dried over Na2SO4, filtered and concentrated to give the title compound.

[0129] LCMS: m / z = 221.1 / 223.1 [M+H] +

[0130] Synthesis of tert-butyl 6-(1-ethoxy-1,3-dioxopentan-2-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate Int 2

[0131]

[0132] Ethyl 2-bromo-3-oxopentanoate (1.1 g, 4.93 mmol), tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (977.5 mg, 27.1 mmol) and K2CO3 (4.1 g, 29.6 mmol) were mixed in acetonitrile (20 mL). The suspension was stirred at room temperature for 30 min. The reaction mixture was filtered. The combined filtrate and washings were concentrated under reduced pressure. The residue was purified by column chromatography (0 - 10% MeOH in dichloromethane) to afford the title compound.

[0133] LCMS (ESI): m / z = 341.2 [M+H] +

[0134] Synthesis of 2-chloro-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide Int 3

[0135]

[0136] To a solution of 2-methyl-4-(trifluoromethyl)aniline (6 g, 34.3 mmol) in dichloromethane (50 mL) at 0 °C was added 2-chloroacetyl chloride (2.87 mL, 36 mmol) and triethylamine (9.1 mL, 65 mmol). The reaction was stirred for 30 min, then warmed to room temperature and stirred for an additional 1.5 h. The mixture was diluted with dichloromethane and washed with water. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was suspended in a small amount of dichloromethane and filtered. The filter cake was washed with dichloromethane and Et2O and dried under vacuum overnight to afford the title compound with acceptable purity. The combined filtrate and washings were concentrated and the residue was purified by column chromatography (50%-100% EtOAc in heptane) to afford the title compound, which was combined with the filter cake.

[0137] LCMS: m / z = 250.1 / 252.1 [M-H] -

[0138] Synthesis of 2-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide Int 4

[0139]

[0140] To a solution of 2-chloro-4-(trifluoromethyl)aniline (5 g, 25.6 mmol) in dichloromethane (50 mL) at 0 °C was added 2-chloroacetyl chloride (2.14 mL, 26.9 mmol) and triethylamine (7.1 mL, 51 mmol). The reaction was stirred for 30 min, then warmed to room temperature and stirred for an additional 1.5 h. The mixture was diluted with dichloromethane and washed with water. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was suspended in a small amount of dichloromethane and filtered. The filter cake was washed with dichloromethane and Et2O and dried under vacuum overnight to afford the title compound with acceptable purity. The combined filtrate and washings were concentrated and the residue was purified by column chromatography (50%-100% EtOAc in heptane) to afford the title compound, which was combined with the filter cake.

[0141] Synthesis of 4-chloro-5-methoxy-6-methylpyrimidine Int 5

[0142]

[0143] A mixture of 4,6-dichloro-5-methoxypyrimidine (25 g, 140 mmol), methylboronic acid (8.8 g, 147 mmol), K3PO4 (74 g, 350 mmol), and Pd(dppf)Cl2.DCM (6.86 g, 8.4 mmol) in DME (100 mL) was stirred at 85 °C for 18 h. The reaction mixture was filtered through a pad of celite and the filtrate was concentrated. The residue was dissolved in EtOAc and washed with saturated aqueous NaHCO3. The organic layer was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (0 - 5% EtOAc in heptane) to afford the title compound.

[0144] 1 H NMR (400 MHz, DMSO-d6) δ 8.66 (s, 1H), 3.86 (s, 3H), 2.50 (s, 3H)

[0145] Synthesis of methyl 5-methoxy-6-methylpyrimidine-4-carboxylate Int 6

[0146]

[0147] To a solution of 4-chloro-5-methoxy-6-methylpyrimidine (14 g, 88.2 mmol) in MeOH (70 mL) was added Pd(dppf)Cl2.DCM (4.32 g, 5.3 mmol) and triethylamine (24.5 mL, 177 mmol). The resulting mixture was stirred at 60 °C under CO (0.4 MPa) for 16 h and then allowed to cool to room temperature. The reaction mixture was filtered and the collected filtrate was concentrated. The residue was dissolved in EtOAc and washed with water. The aqueous layer was back-extracted with EtOAc. The combined organic layers were concentrated and the residue was purified by column chromatography (10% EtOAc in heptane, isocratic) to afford the title compound.

[0148] 1 H NMR (400 MHz, DMSO-d6) δ 8.85 (s, 1H), 3.93 (s, 3H), 3.84 (s, 3H), 1.17 (s, 3H).

[0149] Synthesis of 5-hydroxy-6-methylpyrimidine-4-carboxylic acid Int 7

[0150]

[0151] Methyl 5-methoxy-6-methylpyridine-4-carboxylate (5.5 g, 29 mmol) was dissolved in HBr (40% in water) (23.5 mL, 174 mmol). The resulting mixture was stirred at 40 °C for 10 h. Then HI (23 mL, 174 mmol) was added, and the mixture was stirred for an additional 6 h, and then its pH was adjusted to 3 with 50% NaOH solution at 0 °C. The yellow solid was separated by filtration and then suspended in water (40 mL) and HCl (37% in water) (11 mL, 134 mmol). The resulting suspension was stirred at 60 °C for 2 h and then filtered. The solid was collected and dried under vacuum to afford the title compound.

[0152] 1 1H NMR (400 MHz, DMSO-d6) δ 11.71 (s, 2H), 8.66 (s, 1H), 2.47 (s, 3H).

[0153] Synthesis of tert-butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate Int 8

[0154] Step 1. tert-Butyl 4-(1-ethoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate

[0155]

[0156] Ethyl 2-bromo-3-oxopentanoate (2 g, 8.96 mmol), tert-butyl piperazine-1-carboxylate (977.5 mg, 27.1 mmol) and K2CO3 (9.18 g, 49.3 mmol) were mixed in acetonitrile (40 mL). The suspension was stirred at room temperature for 30 min. The reaction mixture was filtered. The combined filtrate and washings were concentrated under reduced pressure. The residue was purified by column chromatography (0 - 10% MeOH in dichloromethane) to afford the title compound.

[0157] Step 2. tert-Butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0158]

[0159] To a solution of tert-butyl 4-(1-ethoxy-1,3-dioxolan-2-yl)piperazine-1-carboxylate (765.2 mg, 2.33 mmol) in EtOH (4 mL) was added 3-bromo-1H-1,2,4-triazol-5-amine (400 mg, 2.33 mmol) and H3PO4 (282 mg, 2.95 mmol). The resulting mixture was stirred at 85 °C for 24 h and then cooled to room temperature. DIPEA (1.22 mL, 7 mmol) and Boc2O (255 mg, 1.17 mmol) were added and the mixture was stirred at room temperature for an additional 2 h and then quenched with a small amount of water. EtOH was removed under reduced pressure and the residue was dissolved in EtOAc, washed with water, brine, dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 75% EtOAc in heptane) to afford the title compound.

[0160] Step 3. tert-Butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0161]

[0162] To a mixture of tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (125 mg, 0.29 mmol) and 2-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (86.8 mg, 0.32 mmol) in 1,4-dioxane (4 mL) was added KI (48.1 mg, 0.29 mmol) and DIPEA (152 μL, 0.87 mmol). The resulting mixture was stirred at 80 °C for 3.5 h and then cooled to room temperature. The mixture was partitioned between EtOAc and water. The organic layer was washed with brine, dried (Na2SO4), filtered and concentrated. The residue was purified by column chromatography (0 - 50% EtOAc in heptane) to afford the title compound.

[0163] Synthesis of tert-butyl 4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate Int 9.

[0164]

[0165] Int 9 can be prepared using 2-chloro-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide Int3 as a synthetic intermediate according to the same synthetic route as Int 8.

[0166] Synthesis of tert-butyl 3-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 10.

[0167]

[0168] Step 1. Methyl 2-chloro-3-oxopentanoate

[0169]

[0170] SO2Cl2 (13.48 g, 99.89 mmol) was added to a solution of methyl 3-oxopentanoate (10 g, 76.84 mmol) in DCM (100 ml) at 0 °C over 10 min. The reaction was allowed to warm to room temperature and stirred for 16 h. The RM was concentrated under reduced pressure and the residue was dissolved in DCM (100 ml) and washed with water (100 ml), brine (100 ml), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give the title compound as a pale yellow liquid (12 g, 94% yield). Calculated mass for C6H9ClO3 164.0, found: 163.0 (M-H)-ESI.

[0171] 1H NMR (400 MHz, DMSO-d6) δ: 5.62 (s, 1H), 3.76 (s, 3H), 2.50 - 2.48 (m, 2H), 0.99 (t, J = 7.2 Hz, 3H).

[0172] Step 2. Tert-butyl 3-(1-methoxy-1,3-dioxolan-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.

[0173]

[0174] To a solution of methyl 2-chloro-3-oxopentanoate (5.04 g, 30.62 mmol) in dry ACN (25 ml) was added TEA (7.15 g, 70.66 mmol) over 15 min, then tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (5 g, 23.55 mmol) in ACN was added dropwise over 30 min. The reaction was stirred at 60 °C for 16 h. The RM was filtered and washed with EA. Then the filtrate was concentrated under reduced pressure and the residue was dissolved in EA and washed with water, dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography (eluent heptane:EA 100:0 to 90:10) to afford the title compound (4 g, 50% yield). Calculated mass for C17H28N2O5 340.2, found: 339.1 (M-H)-ESI. 1H NMR (400 MHz, DMSO-d6) δ: 4.36 (s, 1H), 3.61 (s, 3H), 3.21 - 3.11 (m, 1H), 2.99 - 2.92 (m, 1H), 2.75 - 2.60 (m, 4H), 2.37 - 2.26 (m, 2H), 1.88 - 1.75 (m, 4H), 1.41 - 1.38 (s, 9H), 0.96 (t, J = 7.3 Hz, 3H).

[0175] Step 3. 3-(2-Bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0176]

[0177] 3-Bromo-1H-1,2,4-triazol-5-amine (843 mg, 5.17 mmol) and tert-butyl (1R,5S)-3-(1-methoxy-1,3-dioxolan-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.6 g, 4.70 mmol) were mixed in EtOH (3 mL). H3PO4 (461 mg, 4.70 mmol) was added. The mixture was stirred at 80 °C under nitrogen for 12 h. The mixture was concentrated in vacuo to remove EtOH, then quenched by addition of saturated aqueous NaHCO3 and extracted with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (silica column, eluent DCM:MeOH 1:0 to 50:1) to give the title compound as a yellow solid (300 mg, 14% yield). Calculated mass for C18H25BrN6O3 452.1 454.1, found: 451.1 453.1 (M-H)-ESI. 1H NMR (400 MHz, DMSO-d6) δ: 13.25 (s, 1H), 4.16 - 4.02 (m, 2H), 3.56 - 3.50 (d, J = 9.8 Hz, 2H), 2.81 - 2.71 (m, 2H), 2.46 - 2.38 (d, J = 9.8 Hz, 2H), 1.93 - 1.80 (m, 4H), 1.44 (s, 9H), 1.21 (t, J = 7.6 Hz, 3H).

[0178] Step 4. 3-(2-Bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0179]

[0180] At room temperature, DIPEA (120 mg, 0.93 mmol) was added to a stirred solution of tert-butyl 3-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (140 mg, 0.31 mmol) and 2-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (84 mg, 0.31 mmol) in 1,4-dioxane (2 mL) and the RM was stirred at 80 °C for 72 h. The RM was concentrated under reduced pressure. The crude product was diluted with EtOAc and water, extracted once with EtOAc and the organic layer was washed with brine, dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography (eluent heptane:EtOAc 70:30 to 30:70) to afford the title compound as a white solid (130 mg, 61% yield). Calculated mass for C27H30BrClF3N7O4 687.1 689.1, found: 686.1 688.1 (M-H)-ESI. 1H NMR (400 MHz, DMSO-d6) δ: 10.34 (s, 1H), 8.11 - 8.06 (d, J = 8.6 Hz, 1H), 7.99 - 7.96 (d, J = 1.7 Hz, 1H), 7.74 - 7.70 (dd, J = 8.7, 1.7 Hz, 1H), 5.29 (s, 2H), 4.15 - 4.07 (m, 2H), 3.61 - 3.57 (m, 2H), 3.02 - 2.92 (m, 2H), 2.49 - 2.43 (m, 2H), 1.92 - 1.84 (m, 4H), 1.45 (s, 9H), 1.20 (t, J = 7.4 Hz, 3H).

[0181] Synthesis of tert-butyl 3-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 11.

[0182]

[0183] Int 11 can be prepared following the same synthetic route as Int 10 using 2-chloro-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide Int 3 as the synthetic intermediate.

[0184] Synthesis of tert-butyl 3-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 12.

[0185]

[0186] Suspend tert-butyl 3-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (186 mg, 0.27 mmol) in 1,4-dioxane (0.5 mL). Add 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (113 mg, 0.54 mmol) and 0.6 M aqueous Na2CO3 solution (0.25 mL) and degas the RM with argon for 10 minutes. Add PdCl2(PPh)2 (19 mg, 0.03 mmol) and stir the RM at 90 °C for 12 hours. Concentrate the RM under reduced pressure and purify the crude product by column chromatography (silica gel column: 12 g silica, eluent DCM:MeOH 100:0 to 97:3) to obtain the crude title compound (72 mg). Calculated mass for C32H37ClF3N7O5 is 691.2, found: 690.1 (M-H)-ESI.

[0187] Synthesis of tert-butyl 3-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 13.

[0188]

[0189] Int 13 can be prepared via the Suzuki coupling of Int 11 and 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane according to the same method as Int 12.

[0190] Synthesis of tert-butyl 3-(5-ethyl-2-(5-fluoro-3,6-dihydro-2H-pyran-4-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 14.

[0191]

[0192] Int 14 can be prepared via the Suzuki coupling of Int 11 and 2-(5-fluoro-3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the same manner as Int 13.

[0193] Synthesis of tert-butyl 3-(5-ethyl-2-(4-fluoro-5,6-dihydro-2H-pyran-3-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 15.

[0194]

[0195] Int 15 can be prepared via the Suzuki coupling of Int 11 and 2-(4-fluoro-5,6-dihydro-2H-pyran-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the same manner as Int 13.

[0196] Synthesis of tert-butyl 3-(2-(3,6-dihydro-2H-thiopyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 16.

[0197]

[0198] Int 16 can be prepared via the Suzuki coupling of Int 11 and 2-(3,6-dihydro-2H-thiopyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the same manner as Int 13.

[0199] Synthesis of tert-butyl 3-(5-ethyl-2-(6-methyl-3,6-dihydro-2H-pyran-4-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 17.

[0200]

[0201] Int 17 can be prepared via Suzuki coupling of Int 11 and 4,4,5,5-tetramethyl-2-(6-methyl-3,6-dihydro-2H-pyran-4-yl)-1,3,2-dioxaborolane in the same manner as Int 13.

[0202] Synthesis of tert-butyl 3-(5-ethyl-2-(2-methyl-3,6-dihydro-2H-pyran-4-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 18.

[0203]

[0204] Int 18 can be prepared via Suzuki coupling of Int 11 and 4,4,5,5-tetramethyl-2-(2-methyl-3,6-dihydro-2H-pyran-4-yl)-1,3,2-dioxaborolane in the same manner as Int 13.

[0205] Synthesis of tert-butyl 3-(5-ethyl-2-(4-methoxycyclopent-1-en-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 19.

[0206]

[0207] Int 19 can be prepared via Suzuki coupling of Int 11 and 2-(4-methoxycyclopent-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the same manner as Int 13.

[0208] Synthesis of tert-butyl 3-(5-ethyl-2-(4-hydroxy-4-methylcyclopent-1-en-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 20.

[0209]

[0210] Int 20 can be prepared via Suzuki coupling of Int 11 and 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopent-3-en-1-ol in the same manner as Int 13.

[0211] Synthesis of tert-butyl 3-(5-ethyl-2-(3-methoxybicyclo[3.1.0]hexan-6-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 21.

[0212]

[0213] Int 21 can be prepared via Suzuki coupling of Int 11 and 2-(3-methoxybicyclo[3.1.0]hexan-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the same manner as Int 13.

[0214] Synthesis of tert-butyl 3-(5-ethyl-2-(3-(methoxycarbonyl)bicyclo[3.1.0]hexan-6-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 22.

[0215]

[0216] Int 22 can be prepared via Suzuki coupling of Int 11 and methyl 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)bicyclo[3.1.0]hexane-3-carboxylate in the same manner as Int 13.

[0217] Synthesis of tert-butyl 3-(2-(3-((tert-butyldimethylsilyl)oxy)bicyclo[3.1.0]hexan-6-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 23.

[0218]

[0219] Int 23 can be prepared via the Suzuki coupling of Int 11 and tert-butyldimethyl((6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)bicyclo[3.1.0]hexan-3-yl)oxy)silane in the same manner as Int 13.

[0220] Synthesis of tert-butyl 3-(2-(3-oxabicyclo[4.1.0]heptan-7-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 24.

[0221]

[0222] Int 24 can be prepared via the Suzuki coupling of Int 11 and 2-(3-oxabicyclo[4.1.0]heptan-7-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the same manner as Int 13.

[0223] Synthesis of tert-butyl 3-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-2-(5-oxaspiro[2.4]heptan-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 25.

[0224]

[0225] Int 25 can be prepared via the Suzuki coupling of Int 11 and 4,4,5,5-tetramethyl-2-(5-oxaspiro[2.4]heptan-1-yl)-1,3,2-dioxaborolane in the same manner as Int 13.

[0226] Synthesis of tert-butyl 3-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-2-(6-oxaspiro[2.5]octan-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 26.

[0227]

[0228] Int 26 can be prepared via Suzuki coupling of Int 11 and 4,4,5,5-tetramethyl-2-(6-oxaspiro[2.5]octan-1-yl)-1,3,2-dioxaborolane in the same manner as Int 13.

[0229] Synthesis of tert-butyl 3-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-2-(5-oxaspiro[2.5]octan-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 27.

[0230]

[0231] Int 27 can be prepared via Suzuki coupling of Int 11 and 4,4,5,5-tetramethyl-2-(5-oxaspiro[2.5]octan-1-yl)-1,3,2-dioxaborolane in the same manner as Int 13.

[0232] Synthesis of tert-butyl 3-(2-(cyclobut-1-en-1-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 28.

[0233]

[0234] Int 28 can be prepared via Suzuki coupling of Int 11 and 2-(cyclobut-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the same manner as Int 13.

[0235] Synthesis of tert-butyl 3-(2-(3,3-difluorocyclobut-1-en-1-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 29.

[0236]

[0237] Int 29 can be prepared via Suzuki coupling of Int 11 and 2-(3,3-difluorocyclobut-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the same manner as Int 13.

[0238] Synthesis of tert-butyl 3-(2-(3-(benzyloxy)cyclobut-1-en-1-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 30.

[0239]

[0240] Int 30 can be prepared via Suzuki coupling of Int 11 and 2-(3-(benzyloxy)cyclobut-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the same manner as Int 13.

[0241] Synthesis of tert-butyl 3-(5-ethyl-2-(3-(methoxycarbonyl)cyclobut-1-en-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Int 31.

[0242]

[0243] Int 31 can be prepared via Suzuki coupling of Int 11 and methyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclobut-2-ene-1-carboxylate in the same manner as Int 13.

[0244] Synthesis of 1-(2,2-difluoroethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine Int 32.

[0245]

[0246] Step 1: 4-Bromo-1,2,3,6-tetrahydropyridine.

[0247] 4-Bromo-3,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (4 g, 15.26 mmol) was suspended in hydrogen chloride in dioxane (30 mL) and the mixture was stirred at room temperature for 2 h. The reaction was concentrated under reduced pressure and used without purification. Calculated mass of C5H8BrN 160.98, 162.98, found: 162.2, 164.2 (M+H) + ESI.

[0248] Step 2: 4-Bromo-1-(2,2-difluoroethyl)-1,2,3,6-tetrahydropyridine.

[0249] 4-Bromo-1,2,3,6-tetrahydropyridine (1 g, 6.17 mmol) was suspended in MeCN (20 mL), 2,2-difluoroethyl trifluoromethanesulfonate (21.72 g, 8.02 mmol) and potassium carbonate (2.56 g, 18.52 mmol) were added, and the mixture was stirred at 80 °C under N2 atmosphere for 16 h. The mixture was diluted with EA and water, extracted with EA and the organic layer was washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The crude product was purified by column chromatography to give the product (958 mg, 68.66% yield). Calculated mass of C7H 10 BrF2N 225.00, 226.99, found: 226.1, 228.2 (M+H) + ESI.

[0250] Step 3: 1-(2,2-difluoroethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine.

[0251] 4-Bromo-1-(2,2-difluoroethyl)-1,2,3,6-tetrahydropyridine (958 mg, 4.24 mmol) was suspended in 1,4-dioxane (20 mL), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.18 g, 4.66 mmol) and potassium acetate (1.25 g, 12.71 mmol) were added and the mixture was degassed with argon for 10 minutes, PdCl2(dppf) (343.51 mg, 0.42 mmol) was added and the mixture was stirred at 90 °C overnight. The mixture was concentrated under reduced pressure and the crude product was purified by column chromatography to give the product (1.05 g, 90.72% yield). C 13 H 22 Calculated mass of BF2NO2 273.17, found: 274.3 (M+H) + ESI.

[0252] Synthesis of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)-1,2,3,6-tetrahydropyridine Int 33.

[0253]

[0254] Int 33 was synthesized from tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate according to the reported procedure (WO 2018067512).

[0255] Synthesis of 5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)-1,2,3,6-tetrahydropyridine Int 34.

[0256]

[0257] Int 34 was prepared from tert-butyl 5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate by the same method as Int 33.

[0258] Synthesis of 7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-(2,2,2-trifluoroethyl)-3-azabicyclo[4.1.0]heptane Int 35.

[0259]

[0260] Int 35 can be prepared from tert-butyl 7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-azabicyclo[4.1.0]heptane-3-carboxylate in the same manner as Int 33.

[0261] Synthesis of 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-N-(2,2,2-trifluoroethyl)bicyclo[3.1.0]hexan-3-amine Int 36.

[0262]

[0263] Int 36 can be prepared from tert-butyl (6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)bicyclo[3.1.0]hexan-3-yl)carbamate in the same manner as Int 33.

[0264] Synthesis of 1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6-(2,2,2-trifluoroethyl)-6-azaspiro[2.5]octane Int 37.

[0265]

[0266] Int 37 can be prepared from tert-butyl 1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6-azaspiro[2.5]octane-6-carboxylate in the same manner as Int 33.

[0267] Synthesis of 1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(2,2,2-trifluoroethyl)-5-azaspiro[2.4]heptane Int 38.

[0268]

[0269] Int 38 can be prepared from tert-butyl 1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-azaspiro[2.4]heptane-5-carboxylate in the same manner as Int 33.

[0270] Synthesis of 1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(2,2,2-trifluoroethyl)-5-azaspiro[2.3]hexane Int 39.

[0271]

[0272] Int 39 can be prepared from tert-butyl 5-azaspiro[2.3]hexane-5-carboxylate 1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) in the same manner as Int 33.

[0273] Synthesis of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-N-(2,2,2-trifluoroethyl)cyclobut-2-en-1-amine Int 40.

[0274]

[0275] Int 40 can be prepared from tert-butyl (3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclobut-2-en-1-yl)carbamate in the same manner as Int 33.

[0276] Preparation Example 1A

[0277] Synthesis of Compound 86

[0278]

[0279] Step 1. 6-(2-Bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester

[0280]

[0281] To a solution of 6-(1-ethoxy-1,3-dioxopentan-2-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (793.2 mg, 2.33 mmol) in EtOH (4 mL) was added 3-bromo-1H-1,2,4-triazol-5-amine (400 mg, 2.33 mmol) and H3PO4 (282 mg, 2.95 mmol). The resulting mixture was stirred at 85 °C for 24 h and then cooled to room temperature. DIPEA (1.22 mL, 7 mmol) and Boc2O (255 mg, 1.17 mmol) were added and the mixture was stirred at room temperature for an additional 2 h and then quenched with a small amount of water. EtOH was removed under reduced pressure and the residue was dissolved in EtOAc, washed with water, brine, dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 75% EtOAc in heptane) to afford the title compound.

[0282] Step 2.6-(2-Bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester

[0283]

[0284] To a mixture of 6-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (110 mg, 0.25 mmol) and 2-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (76.2 mg, 0.28 mmol) in 1,4-dioxane (3 mL) was added KI (41.5 mg, 0.25 mmol) and DIPEA (131 μL, 0.75 mmol). The resulting mixture was stirred at 80 °C for 3.5 h and then cooled to room temperature. The mixture was partitioned between EtOAc and water. The organic layer was washed with brine, dried (Na2SO4), filtered and concentrated. The residue was purified by column chromatography (0 - 50% EtOAc in heptane) to afford the title compound.

[0285] Step 3.6-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester

[0286]

[0287] tert-Butyl 6-(2-bromo-5-ethyl-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (124.2 mg, 184 μmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (58 mg, 276 μmol), K3PO4 (78.5 mg, 368 μmol) and Pd(dppf)Cl2.DCM (15.1 mg, 18.4 μmol) in a mixture of 1,4-dioxane (4 mL) and water (2 mL) were stirred at 80 °C for 8 h. The mixture was cooled to room temperature and diluted with EtOAc. The organic layer was washed with water and brine, dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 100% EtOAc in heptane) to afford the title compound.

[0288] Step 4. N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(2,6-diazaspiro[3.3]heptan-2-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0289]

[0290] TFA (0.25 mL) was added to a solution of tert-butyl 6-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (51.5 mg, 76 μmol) in dichloromethane (1 mL). The mixture was stirred at room temperature for 1 h, then diluted with dichloromethane and neutralized with saturated aqueous NaHCO3. The organic layer was dried (Na2SO4), concentrated and dried under vacuum to afford the title compound.

[0291] Step 5. N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,6-diazaspiro[3.3]heptan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0292]

[0293] To a mixture of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(2,6-diazaspiro[3.3]heptan-2-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (40 mg, 69 μmol) and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (12.8 mg, 82.8 μmol) in DMF (1.5 mL) was added HATU (40.5 mg, 103.5 μmol) and DIPEA (60.2 μL, 345 μmol). The mixture was stirred at room temperature for 2 h and then quenched with water. Saturated aqueous NaHCO3 was added and the mixture was extracted with EtOAc. The organic layer was dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 10% MeOH in dichloromethane) to afford the title compound.

[0294] Preparation Example 1B

[0295] Synthesis of Compound 86 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,6-diazaspiro[3.3]heptan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0296]

[0297] Step 1. tert-Butyl 6-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate

[0298]

[0299] To a solution of tert-butyl 6-(1-ethoxy-1,3-dioxolan-2-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (793.2 mg, 2.33 mmol) in EtOH (4 mL) was added 3-bromo-1H-1,2,4-triazol-5-amine (400 mg, 2.33 mmol) and H3PO4 (282 mg, 2.95 mmol). The resulting mixture was stirred at 85 °C for 24 h and then cooled to room temperature. DIPEA (1.22 mL, 7 mmol) and Boc2O (255 mg, 1.17 mmol) were added and the mixture was stirred at room temperature for an additional 2 h, then it was quenched with a small amount of water. EtOH was removed under reduced pressure and the residue was dissolved in EtOAc, washed with water and brine, dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 75% EtOAc in heptane) to afford the title compound.

[0300] Step 2. tert-Butyl 6-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate

[0301]

[0302] To a mixture of tert-butyl 6-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (110 mg, 0.25 mmol) and 2-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (76.2 mg, 0.28 mmol) in 1,4-dioxane (3 mL) were added KI (41.5 mg, 0.25 mmol) and DIPEA (131 μL, 0.75 mmol). The resulting mixture was stirred at 80 °C for 3.5 h and then cooled to room temperature. The mixture was partitioned between EtOAc and water. The organic layer was washed with brine, dried (Na2SO4), filtered and concentrated. The residue was purified by column chromatography (0 - 50% EtOAc in heptane) to afford the title compound.

[0303] Step 3. tert-Butyl 6-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate

[0304]

[0305] tert-Butyl 6-(2-bromo-5-ethyl-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (124.2 mg, 184 μmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (58 mg, 276 μmol), K3PO4 (78.5 mg, 368 μmol) and Pd(dppf)Cl2.DCM (15.1 mg, 18.4 μmol) in a mixture of 1,4-dioxane (4 mL) and water (2 mL) were stirred at 80 °C for 8 h. The mixture was cooled to room temperature and diluted with EtOAc. The organic layer was washed with water and brine, dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 100% EtOAc in heptane) to afford the title compound.

[0306] Step 4. N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(2,6-diazaspiro[3.3]heptan-2-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0307]

[0308] TFA (0.25 mL) was added to a solution of tert-butyl 6-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (51.5 mg, 76 μmol) in dichloromethane (1 mL). The mixture was stirred at room temperature for 1 h, then diluted with dichloromethane and neutralized with saturated aqueous NaHCO3. The organic layer was dried (Na2SO4), concentrated and dried under vacuum to afford the title compound.

[0309] Step 5. N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,6-diazaspiro[3.3]heptan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0310]

[0311] To a mixture of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(2,6-diazaspiro[3.3]heptan-2-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (40 mg, 69 μmol) and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (12.8 mg, 82.8 μmol) in DMF (1.5 mL) at room temperature were added HATU (40.5 mg, 103.5 μmol) and DIPEA (60.2 μL, 345 μmol). The mixture was stirred at room temperature for 2 h and then quenched with water. Saturated aqueous NaHCO3 was added and the mixture was extracted with EtOAc. The organic layer was dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 10% MeOH in dichloromethane) to afford the title compound.

[0312] Preparation Example 2

[0313] Synthesis of Compound 163

[0314]

[0315] Step 1. (E)-6-(4-(2-((2-Chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(3-methoxyprop-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester

[0316]

[0317] tert-Butyl 6-(2-bromo-5-ethyl-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (200 mg, 296 μmol), (E)-2-(3-methoxyprop-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (88 mg, 444 μmol), K3PO4 (126.3 mg, 592 μmol), and Pd(dppf)Cl2.DCM (24.3 mg, 29.6 μmol) in a mixture of 1,4-dioxane (6 mL) and water (3 mL) were stirred at 80 °C for 8 h. The mixture was cooled to room temperature and diluted with EtOAc. The organic layer was washed with water and brine, dried (Na2SO4), and concentrated. The residue was purified by column chromatography (0 - 100% EtOAc in heptane) to afford the title compound.

[0318] Step 2. (E)-N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-methoxyprop-1-en-1-yl)-7-oxo-6-(2,6-diazaspiro[3.3]heptan-2-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0319]

[0320] TFA (0.25 mL) was added to a solution of (E)-tert-butyl 6-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(3-methoxyprop-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (70 mg, 105 μmol) in dichloromethane (1 mL). The mixture was stirred at room temperature for 1 h, then diluted with dichloromethane and neutralized with saturated aqueous NaHCO3. The organic layer was dried (Na2SO4), concentrated, and dried under vacuum to afford the title compound.

[0321] Step 3. (E)-N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(6-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,6-diazaspiro[3.3]heptan-2-yl)-2-(3-methoxyprop-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0322]

[0323] To a mixture of (E)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-methoxyprop-1-en-1-yl)-7-oxo-6-(2,6-diazaspiro[3.3]heptan-2-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (45 mg, 79.5 μmol) and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (14.7 mg, 95.4 μmol) in DMF (2 mL) were added HATU (46.7 mg, 119.3 μmol) and DIPEA (70 μL, 400 μmol). The mixture was stirred at room temperature for 2 h and then quenched with water. Saturated aqueous NaHCO3 was added and the mixture was extracted with EtOAc. The organic layer was dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 10% MeOH in dichloromethane) to afford the title compound.

[0324] Preparation Example 3

[0325] Synthesis of Compound 71

[0326]

[0327] Step 1. tert-Butyl 4-(1-ethoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate

[0328]

[0329] Ethyl 2-bromo-3-oxopentanoate (2 g, 8.96 mmol), tert-butyl piperazine-1-carboxylate (977.5 mg, 27.1 mmol) and K2CO3 (9.18 g, 49.3 mmol) were mixed in acetonitrile (40 mL). The suspension was stirred at room temperature for 30 min. The reaction mixture was filtered. The combined filtrate and washings were concentrated under reduced pressure. The residue was purified by column chromatography (0 - 10% MeOH in dichloromethane) to afford the title compound.

[0330] Step 2. tert-Butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0331]

[0332] To a solution of tert-butyl 4-(1-ethoxy-1,3-dioxolan-2-yl)piperazine-1-carboxylate (765.2 mg, 2.33 mmol) in EtOH (4 mL) was added 3-bromo-1H-1,2,4-triazol-5-amine (400 mg, 2.33 mmol) and H3PO4 (282 mg, 2.95 mmol). The resulting mixture was stirred at 85 °C for 24 h and then cooled to room temperature. DIPEA (1.22 mL, 7 mmol) and Boc2O (255 mg, 1.17 mmol) were added and the mixture was stirred at room temperature for an additional 2 h and then quenched with a small amount of water. EtOH was removed under reduced pressure and the residue was dissolved in EtOAc, washed with water, brine, dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 75% EtOAc in heptane) to afford the title compound.

[0333] Step 3. tert-Butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0334]

[0335] To a mixture of tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (125 mg, 0.29 mmol) and 2-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (86.8 mg, 0.32 mmol) in 1,4-dioxane (4 mL) was added KI (48.1 mg, 0.29 mmol) and DIPEA (152 μL, 0.87 mmol). The resulting mixture was stirred at 80 °C for 3.5 h and then cooled to room temperature. The mixture was partitioned between EtOAc and water. The organic layer was washed with brine, dried (Na2SO4), filtered and concentrated. The residue was purified by column chromatography (0 - 50% EtOAc in heptane) to afford the title compound.

[0336] Step 4. tert-Butyl 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(1-methyl-2-oxo-1,2-dihydropyridin-4-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0337]

[0338] tert-Butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (200 mg, 302 μmol), 1-methyl-2-oxo-1,2-dihydropyridin-4-ylboronic acid (69.3 mg, 453 μmol), K3PO4 (128.9 mg, 604 μmol) and Pd(dppf)Cl2.DCM (24.8 mg, 30.2 μmol) in a mixture of 1,4-dioxane (6 mL) and water (3 mL) were stirred at 80 °C for 8 h. The mixture was cooled to room temperature and diluted with EtOAc. The organic layer was washed with water and brine, dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 100% EtOAc in heptane) to afford the title compound.

[0339] Step 5. N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(1-methyl-2-oxo-1,2-dihydropyridin-4-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0340]

[0341] TFA (0.25 mL) was added to a solution of tert-butyl 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(1-methyl-2-oxo-1,2-dihydropyridin-4-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (76 mg, 110 μmol) in dichloromethane (1 mL). The mixture was stirred at room temperature for 1 h, then diluted with dichloromethane and neutralized with saturated aqueous NaHCO3. The organic layer was dried (Na2SO4), concentrated and dried under vacuum to afford the title compound.

[0342] Step 6. N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-2-(1-methyl-2-oxo-1,2-dihydropyridin-4-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0343]

[0344] To a mixture of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(1-methyl-2-oxo-1,2-dihydropyridin-4-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (55 mg, 93 μmol) and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (17.2 mg, 111.6 μmol) in DMF (2.5 mL) was added HATU (54.6 mg, 139.5 μmol) and DIPEA (81 μL, 465 μmol). The mixture was stirred at room temperature for 2 h and then quenched with water. Saturated aqueous NaHCO3 was added and the mixture was extracted with EtOAc. The organic layer was dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 10% MeOH in dichloromethane) to afford the title compound.

[0345] Preparation Example 4

[0346] Synthesis of Compound 57

[0347]

[0348] Step 1. tert-Butyl 4-(2-bromo-5-ethyl-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0349]

[0350] To a solution of tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (600 mg, 1.4 mmol) in THF (8 mL) and DMF (2 mL) at 0 °C was added 60% NaH in mineral oil (67.2 mg, 1.68 mmol). The mixture was stirred for 30 min and then (2-(chloromethoxy)ethyl)trimethylsilane (285 μL, 1.61 mmol) was added dropwise. The ice bath was removed and the reaction mixture was allowed to warm to room temperature and stirred for 24 h. Saturated aqueous NaHCO3 was added and the mixture was extracted with dichloromethane. The combined organic layers were dried (Na2SO4) and concentrated to afford the title compound.

[0351] Step 2. tert-Butyl 4-(5-ethyl-7-oxo-2-(3-oxomorpholino)-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0352]

[0353] A mixture of tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (100 mg, 179 μmol), morpholin-3-one (36.4 mg, 360 μmol), N,N'-dimethylethylenediamine (1.6 mg, 18 μmol), CuI (1.7 mg, 9 μmol) and K2CO3 (50 mg, 360 μmol) in toluene was stirred at 110 °C for 12 h. The mixture was cooled to room temperature and poured into water. The resulting mixture was extracted with EtOAc. The organic layer was dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 100% EtOAc in heptane) to afford the title compound.

[0354] Step 3. tert-Butyl 4-(5-ethyl-7-oxo-2-(3-oxomorpholino)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0355]

[0356] TFA (0.5 mL) was added to a solution of tert-butyl 4-(5-ethyl-7-oxo-2-(3-oxomorpholino)-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (45 mg, 77.9 μmol) in dichloromethane (1 mL). The mixture was stirred at 40 °C for 2 h and concentrated. The residue was suspended in dichloromethane (2 mL), and DIPEA (27 μL, 156 μmol) and Boc2O (34 mg, 156 μmol) were added. The resulting mixture was stirred at room temperature for 1.5 h and then quenched with saturated aqueous NaHCO3. The mixture was extracted with dichloromethane, and the organic layer was dried (Na2SO4) and concentrated to afford the title compound.

[0357] Step 4. tert-Butyl 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(3-oxomorpholino)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0358]

[0359] To a mixture of tert-butyl 4-(5-ethyl-7-oxo-2-(3-oxomorpholino)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (92 mg, 206 μmol) in 1,4-dioxane (4 mL) was added KI (34.2 mg, 206 μmol) and DIPEA (108 μL, 618 μmol). The resulting mixture was stirred at 80 °C for 3.5 h and then cooled to room temperature. The mixture was partitioned between EtOAc and water. The organic layer was washed with brine, dried (Na2SO4), filtered and concentrated. The residue was purified by column chromatography (0 - 50% EtOAc in heptane) to afford the title compound.

[0360] Step 5. N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-7-oxo-2-(3-oxomorpholino)-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0361]

[0362] To a solution of tert-butyl 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(3-oxomorpholino)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (40 mg, 58.6 μmol) in dichloromethane (1 mL) was added TFA (0.25 mL). The mixture was stirred at room temperature for 1 h, then diluted with dichloromethane and neutralized with saturated aqueous NaHCO3. The organic layer was dried (Na2SO4), concentrated and dried under vacuum to afford the title compound.

[0363] Step 6. N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-2-(3-oxomorpholino)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0364]

[0365] To a mixture of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-7-oxo-2-(3-oxomorpholino)-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (35 mg, 60 μmol) and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (11.1 mg, 72 μmol) in DMF (1.5 mL) was added HATU (35.2 mg, 90 μmol) and DIPEA (52 μL, 300 μmol). The mixture was stirred at room temperature for 2 h and then quenched with water. Saturated aqueous NaHCO3 was added and the mixture was extracted with EtOAc. The organic layer was dried (Na2SO4) and concentrated. The residue was purified by column chromatography (0 - 10% MeOH in dichloromethane) to afford the title compound.

[0366] Preparation Example 5

[0367] Synthesis of Compound 44

[0368]

[0369] Step 1. tert-Butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0370]

[0371] A mixture of tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (120 mg), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (50 mg), K3PO4 (78 mg), and Pd(dppf)Cl2.DCM (15 mg) in 1,4-dioxane (4 mL) and water (2 mL) was stirred at 80 °C for 8 h. The mixture was cooled to room temperature and diluted with EtOAc. The organic layer was washed with water and brine, dried (Na2SO4), and concentrated. The residue was purified by column chromatography (0 - 100% EtOAc in heptane) to afford the title compound.

[0372] Step 2. tert-Butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4-(prop-2-yn-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0373]

[0374] To a solution of tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (430 mg) in DMF (5 mL) stirred at 0 °C was added NaH (60 mg, 60% in mineral oil). After 30 min, propargyl bromide (0.2 mL) was added. The reaction mixture was allowed to warm gradually to room temperature and stirred for 24 h. Saturated NH4Cl solution was added and the resulting solution was extracted with ethyl acetate (3 x 10 mL). The combined organic layers were dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (0 - 10% MeOH in dichloromethane) to afford tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4-(prop-2-yn-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (260 mg). LC-MS MS m / z [M + H - Boc] + 413

[0375] Step 3. tert-butyl 4-(4-((1-(2-chloro-4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0376]

[0377] To a solution of tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4-(prop-2-yn-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (250 mg) and 1-azido-2-chloro-4-(trifluoromethyl)benzene (110 mg) in DMF (3 mL) were added CuI (10 mol%) and DIPEA (0.07 mL). After 5 minutes, ice-cold water was added to quench the reaction. The resulting solution was extracted with ethyl acetate (3 x 10 mL). The combined organic layers were dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (0 - 10% MeOH in dichloromethane) to afford tert-butyl 4-(4-((1-(2-chloro-4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (200 mg). LC-MS MS m / z [M + H - Boc] + 634

[0378] Step 4. 4-((1-(2-Chloro-4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one

[0379]

[0380] To tert-butyl 4-(4-((1-(2-chloro-4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (190 mg) was added HCl (4 M, 2 mL) in dioxane. After stirring at room temperature for 16 h, it was concentrated in vacuo to give the crude 4-((1-(2-chloro-4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one. LC-MS MS m / z [M + 1] + 590

[0381] Step 5.

[0382]

[0383] To a stirred DMF (2 mL) solution of the above crude 4-((1-(2-chloro-4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one from the above step 3, DIPEA (0.1 mL), N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate (140 mg), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (130 mg) were added. After 24 h at room temperature, it was concentrated in vacuo. The residue was purified on an HPLC preparative column to afford 4-((1-(2-chloro-4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one (20 mg). LC-MS MS m / z [M+1] + 726

[0384] Preparation Example 6

[0385] Synthesis of Compound 97

[0386]

[0387] Synthesis Scheme:

[0388]

[0389] Preparation Example 7

[0390] Synthesis of Compound 187

[0391]

[0392] Step 1: tert-Butyl 3-(1-ethoxy-1,3-dioxopentan-2-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate,

[0393] To a stirred solution of tert-butyl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (5 g, 25.22 mmol) and ethyl 2-chloro-3-oxopentanoate (4.95 g, 27.74 mmol) in ACN (50 mL) was added TEA (7.66 g, 75.66 mmol) at room temperature and the mixture was stirred at 55 °C overnight. The mixture was concentrated under reduced pressure. The crude product was diluted with EA and water, extracted once with EA and the organic layer was washed with brine, dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography (PE:EA 10:1 to 5:1) to afford compound 022-1 (4.5 g, 52% yield).

[0394] C 17 H 28 Calculated mass for N2O5 340.2, found: 341.2 (M+H) + ESI.

[0395] Step 2: tert-Butyl 3-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate, 022-2

[0396] Int-1-2 (1.9 g, 11.66 mmol) and tert-butyl 3-(1-ethoxy-1,3-dioxolan-2-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (4.5 g, 13.22 mmol) were mixed in EtOH (50 mL), and H3PO4 (1.14 g, 11.66 mmol) was added. The mixture was stirred under nitrogen at 80 °C for 2 days. The mixture was concentrated in vacuo to remove EtOH, then quenched by addition of saturated NaHCO3 solution and extracted with EA. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (silica column, eluent DCM:MeOH 20:1 to 10:1) to afford compound 022-2 as a yellow oil (650 mg, 12% yield).

[0397] C 17 H 23 Calculated mass for BrN6O3 438.10, 440.10, found: 339.10, 341.10 (M-Boc+H) ESI.

[0398] Step 3: tert-Butyl 3-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate, 022-3

[0399] At room temperature, DIPEA (529.55 mg, 4.10 mmol) was added to a stirred solution of 002-2 (600 mg, 1.37 mmol) and 2-bromo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (44.83 mg, 1.50 mmol) in 1,4-dioxane (15 mL) and the mixture was stirred at 80 °C for 6 h. The mixture was concentrated under reduced pressure. The crude product was diluted with EA and water, extracted once with EA and the organic layer was washed with brine, dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography (DCM:MeOH 25:1 to 10:1) to afford compound 022-3 (710 mg, 79% yield).

[0400] C 27 H 31 Calculated mass for BrF3N7O4 653.16, 655.16, found: 554.06, 556.04 (M - Boc + H) + ESI.

[0401] Step 4: tert-Butyl 3-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate, 022-4

[0402] 022-3 (750 mg, 1.15 mmol) was suspended in 1,4-dioxane (7 mL), H2O (3.5 mL), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (264.81 mg, 1.26 mmol) and K3PO4 (729.75 mg, 3.44 mmol) were added and the mixture was degassed with argon for 10 minutes, PdCl2(dppf) (46.44 mg, 0.06 mmol) was added and the mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure. The crude product was diluted with EA and water, extracted once with EA and the organic layer was washed with brine, dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography to give compound 022-4 (580 mg, 76.9% yield).

[0403] C 32 H 38 Calculated mass for F3N7O5 657.29, 659.30, found: 558.23, 559.25 (M-Boc+H) + ESI.

[0404] Step 5: 2-(6-(3,6-Diazabicyclo[3.1.1]heptan-3-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, 022-5

[0405] 022-4 (100 mg, 0.15 mmol) was suspended in DCM (2 mL), TFA (1 mL) was added and the RM was stirred at room temperature for 2 h. The RM was concentrated under reduced pressure and used without purification.

[0406] C 27 H 30 Calculated mass for F3N7O3 557.24, found: 558.2, (M+H) + ESI.

[0407] Step 6: 2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide,

[0408] 5-Hydroxy-6-methylpyrimidine-4-carboxylic acid (46.44 mg, 0.30 mmol) was dissolved in DCM (5 mL). HOBT (40.71 mg, 0.30 mmol), EDCI (57.70 mg, 0.30 mmol), pyridine (35.75 mg, 0.45 mmol) and 022-5 (84 mg, 0.15 mmol) were added to the solution, and then the mixture was stirred at room temperature for 2 h. The RM was purified by a reverse-phase column (C18 column) to give the product (60 mg, 57% yield).

[0409] C 33 H 34 Calculated mass for F3N9O5: 693.26, found: 694.25 (M+H) + ESI.

[0410] Preparation Example 8

[0411] Synthesis of Compound 189

[0412]

[0413] Step 1: tert-Butyl 8-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate, 005-1

[0414] Int-2 (300 mg, 0.54 mmol) was suspended in DMF (10 mL). AgBF4 (104.15 mg, 0.54 mmol) and tert-butyl 3,8-diazabicyclo[3.2.1]octane-3-carboxylate (1.14 g, 5.4 mmol) were added and then the mixture was stirred at 140 °C under N2 atmosphere for 2 h. The mixture was filtered, diluted with 10 mL H2O and 10 mL EA, the organic phase was washed with brine and concentrated in vacuo to give the crude product. The crude product was purified by a reverse-phase column (C18 column) to give the product 005-1 as a yellow solid (104 mg, 28% yield).

[0415] C 32 H 37 Calculated mass for ClF3N7O5: 691.25, 693.25, found: 636.2, 638.2 (M-tBu+H) + ESI.

[0416] Step 2: 2-(6-(3,8-Diazabicyclo[3.2.1]octan-8-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide, 005-2

[0417] Add 005-1 (100 mg, 0.14 mmol), hydrogen chloride in dioxane (2 mL) and stir the RM at room temperature for 2 h. Concentrate the RM under reduced pressure and use without purification.

[0418] C 27 H 29 Calculated mass of ClF3N7O3 is 591.20, found: 592.2 (M+H) + ESI.

[0419] Step 3: N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-(3-hydroxypyridinecarbonyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide,

[0420] Dissolve 3-hydroxypicolinic acid (14.10 mg, 0.10 mmol) in DMF (1 mL), add HOBT (13.69 mg, 0.10 mmol), EDCI (19.43 mg, 0.10 mmol), pyridine (12.03 mg, 0.10 mmol) and 005-2 (30 mg, 0.051 mmol) to the solution, then stir at room temperature for 2 h. Purify the RM by reverse phase column (C18 column) to afford the product as a white solid (11 mg, 30% yield).

[0421] C 33 H 32 Calculated mass of ClF3N8O5 is 712.21, 714.21, found: 713.2, 715.4 (M+H) + ESI.

[0422] 11H NMR (400 MHz, DMSO-d6) δ: 10.24 (s, 1H), 8.08 - 8.06 (m, 2H), 7.97 (s, 1H), 7.73 - 7.71 (d, J = 8.0 Hz, 1H), 7.28 (s, 2H), 6.82 (s, 1H), 5.32 (s, 2H), 4.31 - 4.25 (m, 3H), 3.80 (s, 2H), 3.63 (s, 1H), 3.44 - 3.41 (m, 4H), 3.15 - 3.10 (m, 4H), 2.33 - 2.22 (m, 2H), 1.86 (s, 1H), 1.70 (s, 1H), 2.37 (s, 3H), 1.27 - 1.23 (m, 3H).

[0423] Preparation Example 9

[0424] Synthesis of Compound 4

[0425]

[0426] Step 1: Methyl 2-chloro-3-oxopentanoate

[0427]

[0428] SO2Cl2 (13.48 g, 99.89 mmol) was added to a solution of methyl 3-oxopentanoate (10 g, 76.84 mmol) in DCM (100 ml) at 0 °C over 10 min. The reaction was allowed to warm to room temperature and stirred for 16 h. The RM was concentrated under reduced pressure and the residue was dissolved in DCM (100 ml) and washed with water (100 ml), brine (100 ml), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give the title compound as a pale yellow liquid (12 g, 94% yield).

[0429] Calculated mass for C6H9ClO3 164.0, found: 163.0 (M-H)-ESI.

[0430] 1 1H NMR (400 MHz, DMSO-d6) δ: 5.62 (s, 1H), 3.76 (s, 3H), 2.50 - 2.48 (m, 2H), 0.99 (t, J = 7.2 Hz, 3H).

[0431] Step 2: tert-Butyl (1R,5S)-3-(1-methoxy-1,3-dioxolan-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0432]

[0433] To a solution of methyl 2-chloro-3-oxopentanoate (5.04 g, 30.62 mmol) in dry ACN (25 ml) was added TEA (7.15 g, 70.66 mmol) over 15 min, and then tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (5 g, 23.55 mmol) in ACN was added dropwise over 30 min. The reaction was stirred at 60 °C for 16 h. The RM was filtered and washed with EA. Then the filtrate was concentrated under reduced pressure and the residue was dissolved in EA and washed with water, dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography (eluent heptane:EA 100:0 to 90:10) to give the title compound (4 g, 50% yield).

[0434] C 17 H 28 Calculated mass for N2O5 340.2, found: 339.1 (M-H)-ESI.

[0435] 1 1H NMR (400 MHz, DMSO-d6) δ: 4.36 (s, 1H), 3.61 (s, 3H), 3.21 - 3.11 (m, 1H), 2.99 - 2.92 (m, 1H), 2.75 - 2.60 (m, 4H), 2.37 - 2.26 (m, 2H), 1.88 - 1.75 (m, 4H), 1.41 - 1.38 (s, 9H), 0.96 (t, J = 7.3 Hz, 3H).

[0436] Step 3: tert-Butyl 3-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0437]

[0438] 3-Bromo-1H-1,2,4-triazol-5-amine (843 mg, 5.17 mmol) and tert-butyl (1R,5S)-3-(1-methoxy-1,3-dioxolan-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.6 g, 4.70 mmol) were mixed in EtOH (3 mL). H3PO4 (461 mg, 4.70 mmol) was added. The mixture was stirred at 80 °C under nitrogen for 12 h. The mixture was concentrated in vacuo to remove EtOH, then quenched by addition of saturated aqueous NaHCO3 and extracted with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (silica column, eluent DCM:MeOH 1:0 to 50:1) to give the title compound as a yellow solid (300 mg, 14% yield).

[0439] C 18 H 25 Calculated mass for C15H22BrN6O3 452.1 454.1, found: 451.1 453.1 (M-H)-ESI.

[0440] 1 H NMR (400 MHz, DMSO-d6) δ: 13.25 (s, 1H), 4.16 - 4.02 (m, 2H), 3.56 - 3.50 (d, J = 9.8 Hz, 2H), 2.81 - 2.71 (m, 2H), 2.46 - 2.38 (d, J = 9.8 Hz, 2H), 1.93 - 1.80 (m, 4H), 1.44 (s, 9H), 1.21 (t, J = 7.6 Hz, 3H).

[0441] Step 4: tert-Butyl 3-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.

[0442]

[0443] To a stirred solution of tert-butyl 3-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (140 mg, 0.31 mmol) and 2-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (84 mg, 0.31 mmol) in 1,4-dioxane (2 mL) at room temperature was added DIPEA (120 mg, 0.93 mmol) and the RM was stirred at 80 °C for 72 h. The RM was concentrated under reduced pressure. The crude product was diluted with EtOAc and water, extracted once with EtOAc and the organic layer was washed with brine, dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography (eluent heptane:EtOAc 70:30 to 30:70) to afford the title compound as a white solid (130 mg, 61% yield).

[0444] C 27 H 30 Calculated mass for BrClF3N7O4 687.1 689.1, found: 686.1 688.1 (M-H)-ESI.

[0445] 1 H NMR (400 MHz, DMSO-d6) δ: 10.34 (s, 1H), 8.11 - 8.06 (d, J = 8.6 Hz, 1H), 7.99 - 7.96 (d, J = 1.7 Hz, 1H), 7.74 - 7.70 (dd, J = 8.7, 1.7 Hz, 1H), 5.29 (s, 2H), 4.15 - 4.07 (m, 2H), 3.61 - 3.57 (m, 2H), 3.02 - 2.92 (m, 2H), 2.49 - 2.43 (m, 2H), 1.92 - 1.84 (m, 4H), 1.45 (s, 9H), 1.20 (t, J = 7.4 Hz, 3H).

[0446] Step 5: tert-butyl 3-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0447]

[0448] tert-Butyl 3-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (186 mg, 0.27 mmol) was suspended in 1,4-dioxane (0.5 mL). 2-(3,6-Dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (113 mg, 0.54 mmol) and 0.6 M aqueous Na2CO3 solution (0.25 mL) were added and the RM was degassed with argon for 10 minutes. PdCl2(PPh)2 (19 mg, 0.03 mmol) was added and the RM was stirred at 90 °C for 12 hours. The RM was concentrated under reduced pressure and the crude product was purified by column chromatography (silica gel column: 12 g silica, eluent DCM:MeOH 100:0 to 97:3) to afford the crude title compound (72 mg).

[0449] C 32 H 37 Calculated mass of ClF3N7O5 691.2, found: 690.1 (M-H)-ESI.

[0450] Step 6: 2-(6-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

[0451]

[0452] tert-Butyl 3-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (40 mg, 0.06 mmol) was suspended in DCM (2 ml), TFA (1 ml) was added and the RM was stirred at room temperature for 4 h. The RM was concentrated under reduced pressure and used without purification.

[0453] C 27 H 29 Calculated mass of ClF3N7O3 591.2, found: 592.1 (M+H)+ESI.

[0454] Step 7: N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(8-(3-hydroxy-4-methylpyridine-1-carbonyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0455] Dissolve 3-hydroxypicolinic acid (9 mg, 0.07 mmol) in DCM (2 ml). Add pyridine (12 mg, 0.16 mmol), HOBT (14.18 mg, 0.10 mmol), EDCI (20.1 mg, 0.10 mmol), and 2-(6-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (30 mg, 0.05 mmol) to this solution. After stirring for 3 hours, purify the RM by a reverse-phase column (C18 column) to afford the product (12 mg, 33% yield).

[0456] C 34 H 34 Calculated mass for ClF3N8O5: 712.2, found: 713.1 (M+H)+ ESI.

[0457] 1 1H NMR (400 MHz, DMSO-d6) δ: 11.84 (s, 1H), 10.37 (s, 1H), 8.15 - 8.10 (m, 1H), 8.07 - 8.04 (d, J = 8.5 Hz, 1H), 7.99 - 7.95 (d, J = 1.6 Hz, 1H), 7.74 - 7.68 (m, 1H), 7.44 - 7.35 (m, 2H), 6.82 (s, 2H), 5.31 (s, 2H), 5.08 (s, 1H), 4.83 (s, 1H), 4.27 - 4.20 (d, J = 2.5 Hz, 1H) 3.09 - 2.96 (m, 2H), 2.71 - 2.64 (d, J = 8.8 Hz, 1H), 2.63 - 2.57 (d, J = 8.7 Hz, 1H), 2.49 - 2.45 (m, 2H), 2.08 - 1.91 (m, 4H), 1.25 - 1.20 (m, 3H).

[0458] Preparation Example 10

[0459] Synthesis of Compounds 190 and 196

[0460]

[0461] Step 1: tert-Butyl 5-(((trifluoromethyl)sulfonyl)oxy)-2,3,4,7-tetrahydro-1H-azepine-1-carboxylate (003-1)

[0462] A mixture of SM1 (2 g, 9.38 mmol) in THF (30 mL) was stirred at -78 °C, LiHMDS (1.13 mL, 11.25 mmol) was added, the mixture was stirred at -78 °C for 1.0 h, SM2 (4.02 g, 11.25 mmol) was added, and the mixture was stirred at 25 °C for 16 h. TLC (PE:EA = 5:1) showed the detection of a new spot and the complete consumption of the starting material. The reaction was quenched with water (20 mL) and extracted with EA (20 mL x 3). The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The obtained crude residue was purified by silica gel column chromatography using PE / EA (5 / 1) to give the product 003-1 (1.6 g, 49.41% yield) as a yellow oil.

[0463] 1 1H NMR (400 MHz, chloroform-d) δ 5.88 (q, J = 5.5 Hz, 1H), 3.95 (dd, J = 32.5, 5.1 Hz, 2H), 3.56 (dt, J = 21.7, 6.2 Hz, 2H), 2.68–2.49 (m, 2H), 1.94 (dt, J = 13.5, 6.5 Hz, 2H), 1.46 (s, 9H).

[0464] Step 2: tert-Butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3,4,7-tetrahydro-1H-azepine-1-carboxylate (003-2)

[0465] To a solution of 003-1 (500 mg, 1.45 mmol) and B2pin2 (0.38 mL, 1.45 mmol) in 1,4-dioxane (10 mL) were added Pd(dppf)Cl2 (105.94 mg, 0.14 mmol) and AcOK (426.29 mg, 4.34 mmol). The mixture was stirred at 90 °C under N2 for 3 h. TLC (PE:EA = 5:1) showed the detection of a new spot and the complete consumption of the starting material. The solvent was removed. The mixture was diluted with water (10 mL) and extracted with EA (10 mL x 3), and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The obtained crude residue was purified by silica gel column chromatography using PE / EA (5 / 1) to obtain 003-2 (216 mg, 46.15% yield) as a yellow oil, which was examined by 2D NMR.

[0466] 1 1H NMR (400 MHz, chloroform-d) δ 4.20 (dd, J = 14.7, 5.3 Hz, 1H), 4.00 (d, J = 13.4 Hz, 1H), 2.64 (s, 1H), 2.52–2.35 (m, 2H), 1.60 (ddt, J = 21.1, 10.4, 4.2 Hz, 4H), 1.47 (s, 9H), 1.18 (d, J = 1.9 Hz, 12H), 0.96 (dd, J = 8.0, 3.9 Hz, 1H), 0.51 (dd, J = 5.3, 3.9 Hz, 1H)

[0467] Step 3: tert-Butyl 5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3,4,7-tetrahydro-1H-azepine-1-carboxylate (003-3)

[0468] To a solution of 003-2 (100 mg, 0.31 mmol) and SM3 (118.40 mg, 0.22 mmol) in 1,4-dioxane (5 mL) and H2O (0.5 mL) were added K2CO3 (128.26 mg, 0.93 mmol) and Pd(dppf)Cl2 (25.26 mg, 0.03 mmol), and the mixture was stirred at 70 °C under N2 for 3 h. LCMS (XT221416-309-6563-Z-1) showed detection of the desired product and complete consumption of the starting material. The solvent was removed. The mixture was diluted with water (20 mL) and extracted with EA (20 mL x 3), and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (1 / 1) to give the product 003-3 (63 mg, 28.56% yield) as a white solid. MS: at 1.65 min, m / z = 621.3 (M+1, ESI+).

[0469] Step 4: N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(2,5,6,7-tetrahydro-1H-azepin-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (003-4)

[0470] A mixture of 003-3 (63 mg, 0.01 mmol) and TFA (0.2 mL, 1.34 mmol) in DCM (2 mL) was stirred at 25 °C for 1 h. LCMS (XT221416-322-21052-LCMS06) indicated that the desired MS peak was found. The solvent was removed to afford the product 003-4 (69 mg, crude) as a yellow oil. MS: at 1.70 min, m / z = 577.2 (M+1, ESI+).

[0471] Step 5: N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(1-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5,6,7-tetrahydro-1H-azepin-4-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Compound 190)

[0472] To a solution of 003-4 (60 mg, 0.10 mmol) and SM4 (32.05 mg, 0.21 mmol) in DMF (3 mL) was added EDC (39.87 mg, 0.21 mmol), HOBt (28.10 mg, 0.21 mmol) and DIEA (0.09 mL, 0.52 mmol), and the mixture was stirred at 25 °C for 3 h. LCMS (XT221416-337-P1-25473-LCMS02) showed that the desired product was detected and the starting material was completely consumed. The reaction was diluted with water (20 mL) and extracted with EA (30 mL x 3). The organic layer was separated and washed with water (30 mL x 3). The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by preparative HPLC to afford Compound 190 (15.2 mg, 20.5% yield) as a white solid. MS: at 1.49 min, m / z = 713.4 (M+1, ESI+).

[0473] 11H NMR (400 MHz, methanol-d4) δ 8.53 (d, J = 9.0 Hz, 1H), 8.15 (t, J = 7.6 Hz, 1H), 7.81 (d, J = 2.1 Hz, 1H), 7.61 (d, J = 8.7 Hz, 1H), 7.03–6.90 (m, 1H), 6.06–5.76 (m, 1H), 5.48–5.28 (m, 2H), 4.60 (ddd, J = 30.3, 16.5, 6.1 Hz, 1H), 4.31 (q, J = 2.7 Hz, 3H), 3.94–3.60 (m, 4H), 2.96–2.73 (m, 2H), 2.57 (d, J = 49.0 Hz, 7H), 2.40–2.23 (m, 1H), 2.14–1.91 (m, 1H), 1.33–1.18 (m, 3H).

[0474] Step 6: N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(1-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5,6,7-tetrahydro-1H-azepin-4-yl)-7-oxo-2-(tetrahydro-2H-pyran-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Compound 196)

[0475] A solution of 003 (9 mg, 0.01 mmol) and Pd / C (0.00 mL, 0.01 mmol) in IPA (5 mL) was stirred under H2 at 25 °C for 24 h. LCMS (XT221416-361-P1-27380-LCMS02) showed the desired product was detected and the starting material was completely consumed. The solution was filtered and concentrated. The residue was purified by preparative HPLC to give Compound 196 as a white solid (9.87 mg, 20.14% yield). MS: at 1.46 min, m / z = 715.4 (M+1, ESI+).

[0476] 11H NMR (400 MHz, methanol-d4) δ 8.50 (s, 1H), 8.14 (t, J = 7.4 Hz, 1H), 7.81 (s, 1H), 7.62 (d, J = 8.7 Hz, 1H), 5.88 (d, J = 69.5 Hz, 1H), 5.34 (dd, J = 33.2, 17.0 Hz, 2H), 4.60 (d, J = 15.4 Hz, 2H), 4.42–4.22 (m, 1H), 4.04 (dd, J = 38.0, 13.7 Hz, 3H), 3.77 (d, J = 16.1 Hz, 1H), 3.55 (t, J = 11.0 Hz, 2H), 3.10 (d, J = 20.2 Hz, 1H), 2.97–2.75 (m, 2H), 2.50 (s, 4H), 1.95 (d, J = 6.1 Hz, 4H), 1.25 (dt, J = 45.8, 7.8 Hz, 5H).

[0477] Preparation Example 11

[0478] Synthesis of Compound 191

[0479]

[0480] Step 1: 4-Methoxycyclopent-1-ene

[0481] To a solution of cyclopent-3-en-1-ol (5.0 g, 59.44 mmol) and methyl iodide (9.28 g, 65.39 mmol) in THF (50 mL) under ice-cooling was added sodium hydride (1.71 g, 71.33 mmol, 60%) portionwise, and the resulting mixture was stirred overnight at room temperature. The reaction mixture was partitioned by adding water and ether, and the organic layer was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure under ice-cooling to give crude 4-methoxycyclopent-1-ene (1.3 g, 22.28% yield).

[0482] Calculated mass for C6H10O: 98.1; found: MS not shown.

[0483] 1 1H NMR (400 MHz, CD2Cl2) δ 5.669 (s, 1H), 5.468 (s, 1H), 4.061 - 4.027 (m, 1H), 3.184 (s, 3H), 2.473 - 2.454 (d, J = 7.6 Hz, 1H), 2.276 - 2.269 (d, J = 2.8 Hz, 1H), 2.237 - 2.230 (d, J = 2.8 Hz, 1H), 2.157 - 2.111 d, J = 2.8 Hz, 1H).

[0484] Step 2: 2-(3-Methoxybicyclo[3.1.0]hexan-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0485] Under nitrogen, a solution of CrCl2 (9.77 g, 79.48 mmol) and TMEDA (9.24 g, 79.48 mmol) in THF (80 mL) was stirred at room temperature for 1 h. 2-(Diiodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (7.82 g, 19.87 mmol) was added. The mixture was stirred at room temperature for 1 h. 4-Methoxycyclopent-1-ene (1.3 g, 13.25 mmol) was added and the mixture was stirred at 50 °C for 20 h. The reaction was quenched with water and extracted with EA. The organic layers were combined, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with 20 / 1 PE / EA to afford 2-(3-methoxybicyclo[3.1.0]hexan-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane, 200 mg, 6.3% yield.

[0486] Calculated mass for C11H19O3B: 238.1; found: MS not shown.

[0487] 1 H NMR (400 MHz, CD2Cl2) δ 3.796 - 3.760 (m, 1H), 3.103 (s, 3H), 2.077 - 2.062 (m, 2H), 1.921 - 1.895 (m, 1H), 1.703 - 1.664 (m, 2H), 1.351 - 1.342 (m, 1H), 1.152 - 1.147 (m, 1H), 1.187 (s, 12H).

[0488] Step 3: tert-Butyl 4-(2-(3-oxabicyclo[3.1.0]hexan-6-yl)-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

[0489] Under nitrogen, 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester (500 mg, 0.75 mmol), 2-(3-methoxybicyclo[3.1.0]hexan-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (269 mg, 1.13 mmol), cataCXium A Pd G3 (110 mg, 0.15 mmol) and K3PO4 (480 mg, 2.26 mmol) in a mixture of DMF (10 mL) and H2O (1 mL) were stirred at 100 °C for 2 h. The reaction was quenched with water and extracted with EA. The organic layers were combined, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with 1 / 1 PE / EA to afford 4-(2-(3-oxabicyclo[3.1.0]hexan-6-yl)-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester, 70 mg, 4.77% yield.

[0490] Calculated mass for C32H39O5N7F3Cl: 693.1.0; found: 692.1 [M-H], ESI. + , ESI.

[0491] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-methoxybicyclo[3.1.0]hexan-6-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0492] 4-(2-(3-oxabicyclo[3.1.0]hexan-6-yl)-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester (60 mg, 0.08 mmol) in HCl / 1,4-dioxane (5 mL) was stirred at room temperature overnight. LC-MS showed complete conversion. The mixture was concentrated in vacuo. The crude compound was used directly in the next step without further purification.

[0493] Calculated mass for C27H31O3N7F3Cl: 593.0; found: 592.0 [M-H], ESI. + , ESI.

[0494] Step 5: N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3-methoxybicyclo[3.1.0]hexan-6-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide, Compound 191

[0495] 5-Hydroxy-6-methylpyrimidine-4-carboxylic acid (26.24 mg, 0.17 mmol), Py (20.51 mg, 0.26 mmol), HOBT (23.36 mg, 0.17 mmol), and EDCI (33.14 mg, 0.17 mmol) in DCM (2 mL) were stirred at room temperature for 0.5 h. N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-methoxybicyclo[3.1.0]hexan-6-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (60 mg, 0.08 mmol) was added and the mixture was stirred at room temperature for 3 h. LC-MS showed complete conversion. The reaction mixture was diluted with additional DCM (10 mL) and washed with water (10 mL), concentrated, and purified by preparative HPLC (C18 column, ACN / water) to give N-(2-Chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3-methoxybicyclo[3.1.0]hexan-6-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Compound 191), 10 mg, 15.85% yield.

[0496] Calculated mass for C33H35O5N9F3Cl: 730.0; Found: 728.0 [M-H], ESI. + , ESI.

[0497] 11H NMR (400 MHz, CD2Cl2) δ 10.322 (s, 1H), 8.573 (s, 1H), 8.065 - 8.044 (d, J = 8.4 Hz, 1H), 7.974 (s, 1H), 7.740 - 7.714 (m, 1H), 5.252 (s, 2H), 4.530 - 4.500 (d, J = 12 Hz, 1H), 3.717 - 3.681 (m, 2H), 3.501 - 3.445 (m, 2H), 3.173 (s, 3H), 2.973 - 2.946 (m, 4H), 2.803 - 2.777 (d, J = 10.4 Hz, 1H), 2.624 - 2.599 (d, J = 10 Hz, 1H), 2.445 (s, 3H), 2.274 - 2.225 (m, 2H), 1.795 (s, 3H), 1.719 - 1.673 (m, 2H), 1.183 - 1.146 (m, 3H).

[0498] Preparation Example 12

[0499] Synthesis of Compound 192

[0500]

[0501] Step 1: 2-(3-Oxabicyclo[3.1.0]hexan-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0502] Under nitrogen, a solution of CrCl2 (10.52 g, 85.6 mmol, 6.0 equiv) and TMEDA (9.95 g, 85.6 mmol, 6.0 equiv) in THF (50 mL) was stirred at room temperature for 1 h. 2-(Diiodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (8.43 g, 21.4 mmol, 1.5 equiv) was added. The mixture was stirred at room temperature for 1 h. 2,5-Dihydrofuran (1.0 g, 14.27 mmol, 1 equiv) was added and the mixture was stirred at 50 °C for 20 h. The reaction was quenched with water and extracted with EA. The organic layers were combined, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with 20 / 1 PE / EA to afford 2-(3-oxabicyclo[3.1.0]hexan-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane, 1.8 g, 60.85% yield.

[0503] Calculated mass for C11H19O3B: 210.0; found: MS not shown.

[0504] 11H NMR (400 MHz, CD2Cl2) δ 3.693 - 3.673 (d, J = 8 Hz, 2H), 3.574 - 3.553 (d, J = 8.4 Hz, 2H), 1.664 - 1.655 (d, J = 3.6 Hz, 2H), 1.163 (s, 12H).

[0505] Step 2: tert-Butyl 4-(2-(3-oxabicyclo[3.1.0]hexan-6-yl)-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate,

[0506] Under nitrogen, a mixture of tert-butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (500 mg, 0.75 mmol), 2-(3-oxabicyclo[3.1.0]hexan-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (206 mg, 0.98 mmol), cataCXium A Pd G3 (110 mg, 0.15 mmol) and K3PO4 (480 mg, 2.26 mmol) in DMF (10 mL) and H2O (1 mL) was stirred at 100 °C for 2 h. The reaction was quenched with water and extracted with EA. The organic layers were combined, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with 98 / 2 DCM / MeOH to afford tert-butyl 4-(2-(3-oxabicyclo[3.1.0]hexan-6-yl)-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate, 26 mg, 5.17% yield.

[0507] Calculated mass for C30H35O5N7F3Cl: 665.0; found: 664.0 [M-H], ESI. + , ESI.

[0508] 11H NMR (400 MHz, CD2Cl2) δ 10.342 (s, = 1H), 8.078 - 8.057 (d, J = 8.4 Hz, 1H), 7.975 - 7.971 (d, J = 1.6 Hz, 1H), 7.739 - 7.713 (m, 1H), 5.279 (s, 2H), 3.906 - 3.885 (m, 4H), 3.692 - 3.671 (d, J = 8.4 Hz, 2H), 3.406 - 3.353 (m, 2H), 2.955 - 2.937 (m, 4H), 2.649 - 2.622 (d, J = 10.8 Hz, 2H), 2.146 - 2.139 (d, J = 2.8 Hz, 2H), 1.854 - 1.837 (m, 1H), 1.433 (s, 9H), 1.178 - 1.141 (m, 3H).

[0509] Step 3: 2-(2-(3-oxabicyclo[3.1.0]hexan-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

[0510] 4-(2-(3-oxabicyclo[3.1.0]hexan-6-yl)-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester (40 mg, 0.06 mmol) in HCl / 1,4-dioxane (5 mL) was stirred overnight at room temperature. LC-MS showed complete conversion. The mixture was concentrated in vacuo. The crude compound was used directly in the next step without further purification.

[0511] Calculated mass for C25H27O3N7F3Cl: 565.9; Found: 564.9 [M - H], ESI. + , ESI.

[0512] Step 4: 2-(2-(3-oxabicyclo[3.1.0]hexan-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide, Compound 192

[0513] 5-Hydroxy-6-methylpyrimidine-4-carboxylic acid (21.79 mg, 0.14 mmol), Py (16.77 mg, 0.21 mmol), HOBT (19.10 mg, 0.14 mmol), and EDCI (21.70 mg, 0.14 mmol) in DCM (2 mL) were stirred at room temperature for 0.5 h. 2-(2-(3-Oxabicyclo[3.1.0]hexan-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (40 mg, 0.07 mmol) was added, and the mixture was stirred overnight at room temperature. LC-MS showed complete conversion. The reaction mixture was diluted with additional DCM (10 mL) and washed with water (10 mL), concentrated, and purified by preparative HPLC (C18 column, ACN / water) to afford 2-(2-(3-oxabicyclo[3.1.0]hexan-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (Compound 192), 5.6 mg, 11.29% yield.

[0514] Calculated mass for C31H31O5N9F3Cl: 702.0; Found: 700.0 [M-H], ESI. + , ESI.

[0515] 1 H NMR (400 MHz, CD2Cl2) δ 10.342 (s, 1H), 8.503 (s, 1H), 8.071 - 8.050 (d, J = 8.4 Hz, 1H), 7.973 (s, 1H), 7.735 - 7.713 (d, J = 8.8 Hz, 1H), 5.279 (s, 2H), 3.904 - 3.883 (d, J = 8.4 Hz, 2H), 3.731 - 3.670 (m, 4H), 3.598 (s, 2H), 2.973 - 2.960 (m, 3H), 2.802 - 2.775 (d, J = 10.8 Hz, 1H), 2.264 - 2.259 (d, J = 2 Hz, 1H), 2.423 (s, 2H), 2.142 (s, 2H), 1.854 - 1.846 (m, 1H), 1.189 - 1.153 (m, 3H).

[0516] Preparation Example 13

[0517] Synthesis of Compound 195

[0518]

[0519] Step 1: Methyl 2-chloro-3-oxopentanoate

[0520] SO2Cl2 (13.48 g, 99.89 mmol) was added to a solution of methyl 3-oxopentanoate (10 g, 76.84 mmol) in DCM (100 ml) at 0 °C over 10 min. The reaction was allowed to warm to room temperature and stirred for 16 h. The RM was concentrated under reduced pressure and the residue was dissolved in DCM (100 ml) and washed with water (100 ml), brine (100 ml), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to afford the title compound as a pale yellow liquid (12 g, 94% yield).

[0521] Calculated mass for C6H9ClO3 164.0, found: 163.0 (M-H)-ESI.

[0522] 1 H NMR (400 MHz, DMSO-d6) δ: 5.62 (s, 1H), 3.76 (s, 3H), 2.50 - 2.48 (m, 2H), 0.99 (t, J = 7.2 Hz, 3H).

[0523] Step 2: tert-Butyl (1R,5S)-3-(1-methoxy-1,3-dioxolan-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0524] TEA (7.15 g, 70.66 mmol) was added to a solution of methyl 2-chloro-3-oxopentanoate (5.04 g, 30.62 mmol) in dry ACN (25 ml) over 15 min, then tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (5 g, 23.55 mmol) in ACN was added dropwise over 30 min and the reaction was stirred at 60 °C for 16 h. The RM was filtered and washed with EA. Then the filtrate was concentrated under reduced pressure and the residue was dissolved in EA and washed with water, dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography (eluent heptane:EA 100:0 to 90:10) to afford the title compound (4 g, 50% yield).

[0525] C 17 H 28 Calculated mass for C H N2O5 340.2, found: 339.1 (M-H)-ESI.

[0526] 11H NMR (400 MHz, DMSO-d6) δ: 4.36 (s, 1H), 3.61 (s, 3H), 3.21 - 3.11 (m, 1H), 2.99 - 2.92 (m, 1H), 2.75 - 2.60 (m, 4H), 2.37 - 2.26 (m, 2H), 1.88 - 1.75 (m, 4H), 1.41 - 1.38 (s, 9H), 0.96 (t, J = 7.3 Hz, 3H).

[0527] Step 3: tert-Butyl 3-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0528] 3-Bromo-1H-1,2,4-triazol-5-amine (843 mg, 5.17 mmol) and (1R,5S)-tert-butyl 3-(1-methoxy-1,3-dioxolan-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.6 g, 4.70 mmol) were mixed in EtOH (3 mL). H3PO4 (461 mg, 4.70 mmol) was added. The mixture was stirred at 80 °C under nitrogen for 12 h. The mixture was concentrated in vacuo to remove EtOH, then quenched by addition of saturated aqueous NaHCO3 and extracted with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (silica column, eluent DCM:MeOH 1:0 to 50:1) to afford the title compound as a yellow solid (300 mg, 14% yield).

[0529] C 18 H 25 Calculated mass for C15H22BrN6O3 452.1, 454.1, found: 451.1, 453.1 (M-H)-ESI.

[0530] 1 1H NMR (400 MHz, DMSO-d6) δ: 13.25 (s, 1H), 4.16 - 4.02 (m, 2H), 3.56 - 3.50 (d, J = 9.8 Hz, 2H), 2.81 - 2.71 (m, 2H), 2.46 - 2.38 (d, J = 9.8 Hz, 2H), 1.93 - 1.80 (m, 4H), 1.44 (s, 9H), 1.21 (t, J = 7.6 Hz, 3H).

[0531] Step 4: tert-Butyl 3-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.

[0532] To a stirred solution of tert-butyl 3-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (140 mg, 0.31 mmol) and 2-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (84 mg, 0.31 mmol) in 1,4-dioxane (2 mL) at room temperature was added DIPEA (120 mg, 0.93 mmol) and the RM was stirred at 80 °C for 72 h. The RM was concentrated under reduced pressure. The crude product was diluted with EtOAc and water, extracted once with EtOAc and the organic layer was washed with brine, dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography (eluent heptane:EtOAc 70:30 to 30:70) to afford the title compound as a white solid (130 mg, 61% yield).

[0533] C 27 H 30 Calculated mass for C25H27BrClF3N7O4 687.1 689.1, found: 686.1 688.1 (M-H)-ESI.

[0534] 1 1H NMR (400 MHz, DMSO-d6) δ: 10.34 (s, 1H), 8.11 - 8.06 (d, J = 8.6 Hz, 1H), 7.99 - 7.96 (d, J = 1.7 Hz, 1H), 7.74 - 7.70 (dd, J = 8.7, 1.7 Hz, 1H), 5.29 (s, 2H), 4.15 - 4.07 (m, 2H), 3.61 - 3.57 (m, 2H), 3.02 - 2.92 (m, 2H), 2.49 - 2.43 (m, 2H), 1.92 - 1.84 (m, 4H), 1.45 (s, 9H), 1.20 (t, J = 7.4 Hz, 3H).

[0535] Step 5: tert-Butyl 3-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0536] tert-Butyl 3-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (186 mg, 0.27 mmol) was suspended in 1,4-dioxane (0.5 mL). 2-(3,6-Dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (113 mg, 0.54 mmol) and 0.6 M aqueous Na2CO3 solution (0.25 mL) were added and the RM was degassed with argon for 10 minutes. PdCl2(PPh)2 (19 mg, 0.03 mmol) was added and the RM was stirred at 90 °C for 12 hours. The RM was concentrated under reduced pressure and the crude product was purified by column chromatography (silica gel column: 12 g silica, eluent DCM:MeOH 100:0 to 97:3) to give the crude title compound (72 mg).

[0537] C 32 H 37 Calculated mass of C19H18ClF3N7O5 is 691.2, found: 690.1 (M-H)-ESI.

[0538] Step 6: 2-(6-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

[0539] tert-Butyl 3-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (40 mg, 0.06 mmol) was suspended in DCM (2 ml), TFA (1 ml) was added and the RM was stirred at room temperature for 4 h. The RM was concentrated under reduced pressure and used without purification.

[0540] C 27 H 29 Calculated mass of C17H17ClF3N7O3 is 591.2, found: 592.1 (M+H)+ESI.

[0541] Step 7: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(8-(3-hydroxy-4-methylpyridine-1-carbonyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0542] Dissolve 5-hydroxy-6-methyl-pyrimidine-4-carboxylic acid (18 mg, 0.12 mmol) in DCM (4 ml). Add pyridine (14 mg, 0.18 mmol), HOBT (16 mg, 0.11 mmol), EDCI (23 mg, 0.11 mmol), 2-(6-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (35 mg, 0.06 mmol) to this solution. After stirring for 3 h, purify the RM by a reverse-phase column (C18 column) to afford the product Compound 195 (15 mg, 34% yield).

[0543] C 34 H 34 Calculated mass for ClF3N8O5 is 726.2, found: 727.1 (M+H)+ESI.

[0544] 1 1H NMR (400 MHz, DMSO-d6) δ: 11.75 (s, 1H), 10.39 (s, 1H), 8.62 (s, 1H), 8.08 - 8.04 (d, J = 8.5 Hz, 1H), 7.99 - 7.97 (s, 1H), 7.74 - 7.70 (d, J = 8.5 Hz, 1H), 6.82 (m, 1H), 5.32 (s, 2H), 5.15 (s, 1H), 4.85 - 4.81 (d, J = 5.7 Hz, 1H), 4.26 - 4.23 (m, 2H), 3.79 (t, J = 5.3 Hz, 2H), 3.72 (t, J = 10.6 Hz, 2H), 3.07 - 3.98 (m, 2H), 2.73 - 2.61 (m, 2H), 2.46 (s, 4H), 2.06 - 1.92 (m, 4H), 1.28 - 1.21 (m, 5H).

[0545] Preparation Example 14

[0546] Synthesis of Compound 202

[0547]

[0548] Step 1: tert-Butyl 3-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate, 139-4

[0549] Suspend 139-3 (100 mg, 0.22 mmol) in DMF (5 mL), add H2O (0.5 mL), 2-(4-methoxycyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (105.06 mg, 0.44 mmol) and Na2CO3 (70.14 mg, 0.66 mmol), and degas the mixture with argon for 10 minutes. Add PdCl2(PPh3)2 (15.48 mg, 22.06 μmol) and stir the mixture at 90 °C overnight. Concentrate the mixture under reduced pressure, and dilute the crude product with DCM and water, extract once with DCM, and wash the organic layer with brine, dry over Na2SO4 and concentrate under reduced pressure. Purify the product by column chromatography to obtain the compound (120 mg, 77.74% yield).

[0550] C 35 H 44 Calculated mass of C29H38F3N7O5 is 699.34, found: 644.3 (M-tBu + H) + ESI.

[0551] Step 2: 2-(6-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, 139-5

[0552] Suspend 139-4 (100 mg, 0.22 mmol) in DCM (5 mL), add hydrogen chloride in dioxane (2 mL) and stir the mixture at room temperature for 2 h. Concentrate the RM under reduced pressure and use without purification.

[0553] C 30 H 36 Calculated mass of C26H33F3N7O3 is 599.28, found: 566.2, 568.2 (M + H) + ESI.

[0554] Step 3: 2-(5-Ethyl-6-(8-(3-hydroxypyridine-2-carbonyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide. Dissolve 139-5 (100 mg, 0.17 mmol) in DCM (5 mL), add 3-hydroxypyridine-2-carboxylic acid (23.20 mg, 0.17 mmol), EDCI (63.94 mg, 0.33 mmol), and HOBT (45.07 mg, 0.33 mmol) to the solution. Stir at room temperature for 2 h, and purify the RM by a reverse-phase column (C18 column) to afford the product HBP-109-LY-NV-139 (38 mg, 31.62% yield). C 36 H 39 Calculated mass of F3N8O5: 720.30, found: 719.4 (M-H) - ESI. 1 H NMR (400 MHz, DMSO-d6) δ: 11.88 (br, 1H), 10.01 (s, 1H), 8.13 - 8.12 (m, 1H), 7.62 (s, 1H), 7.72 (d, J = 8.4 Hz, 1H), 7.53 (d, J = 8.4 Hz, 1H), 7.43 - 7.36 (m, 2H), 6.73 (s, 1H), 5.23 (s, 2H), 5.11 - 5.10 (m, 1H), 4.83 - 4.82 (m, 1H), 3.73 - 3.70 (m, 2H), 3.52 - 3.50 (m, 1H), 3.27 (s, 3H), 3.04 - 3.02 (m, 2H), 2.68 - 2.59 (m, 5H), 2.36 (s, 3H), 2.03 - 2.02 (m, 1H), 1.96 - 1.93 (m, 5H), 1.70 - 1.66 (m, 1H), 1.27 - 1.23 (m, 3H).

[0555] Preparation Example 15

[0556] Synthesis of Compound 203

[0557]

[0558] Step 1: tert-Butyl 3-(2-(1-(2,2-difluoroethyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate, 138-1

[0559] Suspend 3 (390 mg, 0.58 mmol) in DMF (10 mL), add water (1 mL), 1-(2,2-difluoroethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine (239.01 mg, 0.88 mmol) and Na2CO3 (185.50 mg, 1.75 mmol), and degas the mixture with argon for 10 minutes. Add PdCl2(PPh3)2 (40.95 mg, 0.06 mmol) and stir the mixture at 90 °C for 8 h. Concentrate the mixture under reduced pressure and purify the crude product by reverse-phase column (C18 column) to obtain 138-1 (178 mg, 41.53% yield). C 35 H 43 Calculated mass of C25H31F5N8O4 is 734.33, found: 733.51 (M-H) - ESI.

[0560] Step 2: 2-(6-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-(1-(2,2-difluoroethyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, 138-2

[0561] Suspend 138-1 (55 mg, 0.07 mmol) in hydrogen chloride in dioxane (5 mL) and stir the mixture at room temperature for 2 h. Concentrate the RM under reduced pressure and use without further purification. C 30 H 35 Calculated mass of C23H28F5N8O2 is 634.28, found: 635.36 (M+H) + ESI.

[0562] Step 3: 2-(2-(1-(2,2-Difluoroethyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-ethyl-6-(8-(3-hydroxypyridine-2-carbonyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, HBP-109-LY-NV-138

[0563] Dissolve 138-2 (47 mg, 0.07 mmol), 3-hydroxypyridine-2-carboxylic acid (20.60 mg, 0.15 mmol), EDCI (28.39 mg, 0.15 mmol), HOBT (20.01 mg, 0.15 mmol) and pyridine (17.57 mg, 0.22 mmol) in DCM (3 mL), and then stir at room temperature for 2 h. Purify the RM by a reverse-phase column (C18 column) to afford the product HBP-109-LY-NV-138 (18 mg, 32.16% yield). C 36 H 38 Calculated mass for F5N9O4: 755.30, found: 756.41 (M+H) + ESI. 1 H NMR (400 MHz, DMSO-d6) δ: 11.83 (br, 1H), 10.00 (s, 1H), 8.13 - 8.11 (m, 1H), 7.71 (d, J = 8.4 Hz, 1H), 7.62 (s, 1H), 7.53 (d, J = 8.4 Hz, 1H), 7.40 - 7.38 (m, 2H), 6.75 (s, 1H), 6.30 - 6.01 (m, 1H), 5.23 (s, 2H), 5.09 - 5.08 (m, 1H), 4.83 - 4.82 (m, 1H), 3.73 - 3.70 (m, 2H), 3.29 - 3.28 (m, 2H), 3.05 - 3.02 (m, 2H), 2.86 - 2.82 (m, 2H), 2.78 - 2.75 (m, 2H), 2.66 - 2.59 (m, 2H), 2.35 (s, 3H), 2.02 - 1.99 (m, 3H), 1.27 - 1.23 (m, 4H).

[0564] The compounds in the following table were synthesized according to the above examples or routes similar to the above examples.

[0565] Table 1

[0566]

[0567]

[0568]

[0569]

[0570]

[0571]

[0572]

[0573]

[0574]

[0575]

[0576]

[0577]

[0578]

[0579]

[0580]

[0581] Biometrics

[0582] WRN Helicase Assay

[0583] To evaluate the inhibitory properties of the compounds of the present invention against WRN helicase activity, a fluorescence assay using fork DNA was established. In the helicase assay, internally produced His-WRN was used 527-1072Use the fluorescent fork DNA prepared by annealing OLIGOA-BHQ2 (TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCGTACCCGATGTGTTCGTTC-BHQ2) and OLIGOB-TAMRA (TAMRA-GAACGAACACATCGGGTACGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT) as the substrate. Prepare serial 2.5-fold dilutions of the test compound at 7 gradient points with DMSO and then further dilute 1.5 μl of each concentration into 48.5 μl of assay buffer (25 mM Tris-HCl (pH 8.0), 50 mM NaCl, 2 mM MgCl2, 1 mM DTT, 0.05% Tween-20, and 2.5 μg / ml BSA). Transfer 5 μl of 45 nM WRN protein and 5 μl of each diluted compound solution to a 384-well assay plate and pre-incubate at room temperature for 30 minutes. Add each concentration of the compound in triplicate to the plate. Controls are included in the test to validate the assay. Instead of the compound solution, add assay buffer containing 3% DMSO to the positive control (no inhibition), and add no protein to the negative control (maximum inhibition). Dilute the fork DNA and ATP to 300 nM and 6 mM, respectively, with the assay buffer. To initiate the reaction, add 5 μl of the dsDNA and ATP solution to the assay plate. After incubating at room temperature for 60 minutes, transfer the plate to a Victor Nivo multimode plate reader (PerkinElmer, Waltham, MA) and monitor the fluorescence output. Plot the dose-response curve using GraphPad Prism 9 (GraphPad Software, San Diego, CA) to obtain the IC 50 。

[0584] WRN ATPase Assay

[0585] To evaluate the inhibitory properties of the compounds of the present invention on DNA-dependent WRN ATPase activity, an ATPase assay using an ADP-Glo assay kit (Promega, Madison, Wisconsin) was established. In the ATPase assay, the same WRN protein and DNA substrate as described in the helicase assay were used. The compounds in serial dilution in DMSO were the same as in the helicase assay. In further dilution, 2 μl of each concentration was transferred to 48 μl of assay buffer. 5 μl of 100 nM WRN protein and 2.5 μl of each diluted compound solution were pre-incubated in a 384-well assay plate at room temperature for 30 minutes. All test wells were set up in triplicate. Thereafter, 2.5 μl of assay buffer containing 400 nM DNA substrate and 4 mM ATP was added to each well and incubated for 60 minutes. A protein-free control (maximum inhibition) and a compound-free control (no inhibition) were included in the test. To stop the reaction and deplete excess ATP, 10 μl of ADP-Glo TM reagent from the assay kit was added and incubated for 40 minutes. Then, 20 μl of ATP detection reagent was added. After incubation for 30 minutes, luminescence was measured using a Victor Nivo multimode plate reader (PerkinElmer, Waltham, Massachusetts). A dose-response curve was generated by GraphPad Prism 9 (GraphPad Software, San Diego, California) to obtain the IC 50 . Table 1A shows the data of the selected compounds in the above test.

[0586] Table 1A

[0587] Compound Number IC50 (nM) 186 13

[0588] Table 2

[0589]

[0590]

Claims

1. A compound of formula (I)-(XI), or a tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein ring A and ring B are each independently selected from aryl or heteroaryl; Y1 is CH, NH, N, O or S; Y2 is CH, NH, N, O or S; Y3 is CH, NH, N, O or S; X1 and X2 are each independently CH or N; M1, M2, M5, M6 are each independently CH, NH, N, O or S; M3 and M4 are each independently -NR a , -CR a R b ; R a and R b each independently is selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, and hydroxyalkyl; is a single bond or a double bond, provided that are not both double bonds at the same time; R1, R3 and R4 are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; Two R2 together with the atoms to which they are attached form a C3-C 10 cycloalkyl or C3-C 10 heterocyclic group, R5 and R6 are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; or R5 and R6 together with the atoms to which they are attached form a C3-C 10 cycloalkyl, C3-C 10 heterocycloalkyl, C6-C 14 aryl or a C5-C 12 heteroaryl containing 1 to 4 heteroatoms selected from the group consisting of O, S, and N; R7 is selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; or two R7 together with the atoms to which they are attached form a C3-C 10 cycloalkyl or C3-C 10 heterocyclic group, L1 is a key, O, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl, -C3-C 10 cycloalkyl-CONH-, -C1-C6 alkyl-C3-C 10 cycloalkyl-CONH, -C1-C6 haloalkyl-NH-, -C3-C 10 -heterocyclic-NH-alkyl, -C3-C 10 cycloalkyl-NH-, -C1-C6 alkyl-C(NH)-NH, -C6-C 10 heteroaryl-C1-C6 alkyl, -S(O)2-NH- and -C1-C6 alkyl-S(O)2-NH-; L2 is a key, O, -C(O), -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl and -CON(R c )-; R c selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; m is 0, 1, 2, 3 or 4; p is 0, 1, 2 or 3; q is 0, 1, 2 or 3; r is 0, 1, 2 or 3; s is 2 or 3; t is 0, 1, 2 or 3; w is 0, 1, 2 or 3; and n is 0, 1, 2 or 3.

2. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, the compound having formula (XII)-(XIII), (XIX)-(XXI) R8 is C3-C 12 fused cycloalkyl, C 2- C 10 fused heterocyclic group, wherein the fused cycloalkyl group and the fused heterocyclic group are each optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, oxo group; R9 is a bridging C3-C 10 cycloalkyl or a bridging C2-C 10 heterocyclic group, wherein the bridging cycloalkyl and bridging heterocyclic group are each optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxy, C 1-5 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, oxo, C 1-6 haloalkyl, C 3-5 substituents of cycloalkyl; L6 is a bridging C3-C 10 cycloalkyl or bridging C2-C 10 heterocyclic group, wherein the bridging cycloalkyl and bridging heterocyclic groups are each optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, oxo, C 1-6 haloalkyl, C 3-5 substituents of cycloalkyl; Y1, Y2, Y3, M1, M2, L2, X1, X2, R1, R3, R4, m, p, q, r, t are as defined in claim 1.

3. The compound according to claim 1, or a tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, the compound having formula (XXII)-(XXIII) each X is independently selected from F, Cl, Br, I; Y1, Y2, Y3, M1, M2, L2, X1, X2, R1, R3, R7, m, n, p, r, t, w are as defined in claim 1.

4. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein ring A is a C6-C 14 aryl or a C5-C containing 1 to 4 heteroatoms selected from the group consisting of O, S, and N 12 heteroaryl.

5. The compound according to claim 1 or 2 or 3, or a tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R1, R3 and R4 are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 hydroxyalkyl and C2-C6 alkenyl; Preferably, R1, R3 and R4 are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, -CF3.

6. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein two R2 together with the atoms to which they are attached form a bridged C3-C 10 cycloalkyl or a bridged C3-C 10 heterocyclyl; Preferably, represent 7. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R5 and R6 are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl, and C1-C3 hydroxyalkyl; or R5 and R6 together with the atoms to which they are attached form 8. The compound according to claim 1 or 3, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R7 is selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl, and C1-C3 hydroxyalkyl; or two R7 together with the atoms to which they are attached form a bridging C3-C 10 cycloalkyl or bridging C3-C 10 heterocyclic group, preferably denote 9. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R a and R b are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl, and C1-C3 hydroxyalkyl.

10. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein L1 is a bond, O, C1-C3 alkyl, C2-C3 alkenyl, -C1-C3 alkyl-NH-, -C2-C3 alkenyl-NH-, C2-C3 alkynyl, C1-C3 alkoxy, C1-C3 alkylthio, C1-C3 haloalkyl, -C3-C6 cycloalkyl-CONH-, -C1-C3 alkyl-C3-C6 cycloalkyl-CONH, -C1-C3 haloalkyl-NH-, -C3-C6-heterocyclo-NH-alkyl, -C3-C6 cycloalkyl-NH-, -C1-C6 alkyl-C(NH)-NH, -C6-C 10 heteroaryl-C1-C6 alkyl, -S(O)2-NH-, and -C1-C6 alkyl-S(O)2-NH-.

11. The compound according to claim 1 or 2 or 3, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein L2 is a bond, O, -C(O), -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C3 alkyl-NH-, -C2-C3 alkenyl-NH-, C2-C3 alkynyl, C1-C3 alkoxy, C1-C3 alkylthio, C1-C3 haloalkyl, and -CON(R c )-; R c is selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl, and C1-C3 hydroxyalkyl.

12. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein ring A is selected from the group consisting of:

13. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein ring B is selected from the group consisting of phenyl, pyridyl, pyrrolyl, or pyrimidinyl.

14. The compound according to claim 2, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein is 15. The compound according to claim 2, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein in formula (XIII), R8 is selected from the group consisting of:

16. The compound according to claim 2, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R9 is R10 is C 1-5 alkyl, C 3-5 cycloalkyl.

17. The compound according to claim 2, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein L6 is or L6-L2 is 18. The compound according to claim 3, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein in formula (XXII), X attached to the phenyl is Cl, and / or CH n X 3-n is CF3; in formula (XXIII), CH n X 3-n is CF3.

19. A compound selected from the group consisting of: or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

20. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to any one of claims 1 to 19, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, and a pharmaceutically acceptable carrier.

21. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of the compound according to any one of claims 1-19, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or the pharmaceutical composition according to claim 20.

22. A method of treating a disorder or disease in a subject that is treatable by WRN inhibition, the method comprising administering to the subject a therapeutically effective amount of the compound according to any one of claims 1 to 19, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or the pharmaceutical composition according to claim 20.

23. A method for treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound as described in any one of claims 1-19, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a pharmaceutical composition as described in claim 20, wherein the cancer is selected from the group consisting of: microsatellite instability-high (MSI-H) cancer or mismatch repair-deficient (dMMR) cancer, preferably the microsatellite instability-high (MSI-H) cancer or mismatch repair-deficient (dMMR) cancer is selected from colorectal cancer, gastric cancer, prostate cancer, endometrial cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, renal cancer, and ovarian cancer.

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