Piperidone derivative as well as preparation method and medical application thereof

Abstract

The invention discloses a piperidone compound shown in a formula (I), a preparation method thereof, a pharmaceutical composition containing the piperidone compound and medical application of the piperidone compound to treatment of diseases. # imgabs0 #

Description

Technical Field

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

[0002] A large number of observations have emphasized the potential of MK2 as a drug target. Mitogen-activated protein kinase-activated protein kinase 2 (MAPKAP K2 or MK2) mediates a variety of p38 MAPK-dependent cellular responses. MK2 is an important intracellular regulator of cytokine production, such as tumor necrosis factor α (TNFα), interleukin 6 (IL-6), and interferon γ (IFNγ), which are associated with many acute and chronic inflammatory diseases, such as rheumatoid arthritis and inflammatory bowel disease. MK2 is present in the nucleus of unstimulated cells and, upon stimulation, it translocates to the cytoplasm and phosphorylates and activates filaggrin and HSP27. MK2 is also associated with heart failure, cerebral ischemic injury, stress resistance regulation, and the production of TNF-β.

[0003] Many diseases are associated with abnormal cellular responses triggered by protein kinase-mediated events as described above. These diseases include, but are not limited to, autoimmune diseases, inflammatory diseases, bone diseases, metabolic diseases, neurological and neurodegenerative diseases, cancer, cardiovascular diseases, allergies and asthma, Alzheimer's disease, and hormone-related diseases. Therefore, there is a need for compounds that inhibit mitogen-activated protein kinase-activated protein kinase 2 (MK2, MAPKAPK2). Summary of the Invention

[0004] On the one hand, the present invention provides a compound of formula (I):

[0005]

[0006] 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,

[0007] X1, X2, X3, and X4 are each independently selected from a bond, CH, N, O, or S;

[0008] Y1, Y2, Y3, Y4, Y5, and Y6 are each independently selected from a bond, CH, N, O, or S;

[0009] M1, M2, M3, M4, M5, and M6 are each independently selected from a bond, CH, N, O, or S;

[0010] L is a bond, NH, O, S, alkyl, alkoxy, alkyl-N- or -CONH-;

[0011] provided that, a) when L is -CONH-, is a 5-membered ring; b) when L is a bond, is a 5-membered ring;

[0012] R1, R2, R3, R4 and R5 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, hydroxyalkyl, cyano-alkyl, oxo, -CONH2, alkenyl-C(O)NH-, alkynyl-NH-, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, where each of said alkyl, haloalkyl, hydroxyalkyl, oxo, cycloalkyl, heterocycloalkyl, aryl and heteroaryl is independently unsubstituted or substituted at each occurrence by one or more substituents selected from halogen, amino, nitro, cyano, hydroxy, alkyl, alkoxy, haloalkyl, hydroxyalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl;

[0013] or R2 and R3 together with the C atom to which they are attached form a heterocycloalkyl, where said heterocycloalkyl is optionally substituted by one or more substituents selected from halogen, amino, nitro, cyano, hydroxy, alkyl, alkoxy, haloalkyl, hydroxyalkyl, cycloalkyl and heterocycloalkyl;

[0014] t is 0, 1, 2 or 3; and

[0015] s is 0, 1, 2 or 3.

[0016] In some embodiments, when X1, X2, X3 or X4 is selected from the heteroatom N, is saturated or unsaturated and has hydrogen substituents to satisfy the valency;

[0017] In some embodiments, a compound of formula (I-0), or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, is a compound of formula (I):

[0018]

[0019] or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein,

[0020] X1, X2, X3 and X4 are each independently selected from a bond, CH, N, NH, O or S;

[0021] when X1, X2, X3 or X4 is selected from the heteroatom N, is saturated or unsaturated and has hydrogen substituents to satisfy valency; Y1, Y2, Y3, Y4, Y5 and Y6 are each independently selected from a bond, CH, N, O or S;

[0022] M1, M2, M3, M4, M5 and M6 are each independently selected from a bond, CH, N, O or S;

[0023] L is a bond, NH, O, S, alkyl, alkoxy, alkyl-N- or -CONH-;

[0024] provided that, a) when L is -CONH-, is a 5-membered ring; b) when L is a bond, is a 5-membered ring;

[0025] R1, R2, R3, R4 and R5 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, hydroxyalkyl, cyano-alkyl, oxo, -CONH2, alkenyl-C(O)NH-, alkynyl-NH-, -C(O)O-alkyl, -C(O)NH-alkyl, cycloalkyl, heterocyclic group, aryl and heteroaryl, wherein each of said alkyl, haloalkyl, hydroxyalkyl, oxo, cycloalkyl, heterocyclic group, aryl and heteroaryl is independently unsubstituted or substituted by one or more substituents selected from deuterium, halogen, amino, nitro, cyano, hydroxy, alkyl, alkoxy, haloalkyl, hydroxyalkyl, cycloalkyl, heterocyclic group, aryl and heteroaryl;

[0026] or R2 and R3 together with the C atom to which they are attached form a heterocyclic group, wherein said heterocyclic group is optionally substituted by one or more substituents selected from halogen, amino, nitro, cyano, hydroxy, alkyl, alkoxy, haloalkyl, hydroxyalkyl, cycloalkyl and heterocyclic group;

[0027] t is 0, 1, 2 or 3; and

[0028] s is 0, 1, 2 or 3.

[0029] In some embodiments, L is a bond, NH, O, S, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkyl-N- or -CONH-.

[0030] In preferred embodiments, L is a bond, NH, O, S, C 1-3 alkyl, C 1-3 alkoxy, C 1-3 alkyl-N- or -CONH-.

[0031] In some embodiments, R1 is selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group, -CONH2.

[0032] In preferred embodiments, R1 is selected from hydrogen, deuterium, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl.

[0033] In some embodiments, each of R2 and R3 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group, -CONH2.

[0034] In preferred embodiments, each of R2 and R3 is independently selected from hydrogen, deuterium, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl.

[0035] In some embodiments, R2 and R3, together with the C atom to which they are attached, form a 4- to 6-membered heterocyclic group containing 1, 2, or 3 heteroatoms selected from N, O, or S.

[0036] In preferred embodiments, R2 and R3, together with the C atom to which they are attached, form

[0037] In some embodiments, R4 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group, -CONH2;

[0038] In preferred embodiments, R4 is independently selected from hydrogen, deuterium, C 1-3 alkyl, C 1-3 haloalkyl, C1-3 alkoxy, C 1-3 hydroxyalkyl, amino, F, Cl or Br.

[0039] In some embodiments, R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 2-6 cyano-alkyl, oxo, -CONH2, C 3-6 alkenyl-C(O)NH-, C 2-6 alkynyl-NH-, -C(O)O-C 1-4 alkyl, -C(O)NH-C 1-4 alkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 6-10 aryl, a 5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O, wherein each of said alkyl, haloalkyl, hydroxyalkyl, cycloalkyl, heterocyclic group, oxo, aryl and heteroaryl is optionally substituted at each occurrence by one or more substituents selected from deuterium, halogen, amino, nitro, cyano, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 6-10 aryl and a 5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O;

[0040] In preferred embodiments, R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 2-4 cyano-alkyl, C 2-4 alkoxyalkyl, oxo, -CONH2, C 3-6 alkenyl-C(O)NH-, C 2-4 alkynyl-NH-, -C(O)O-C 1-4 alkyl, -C(O)NH-C 1-4 alkyl,

[0041] More preferably, R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 2-4 cyano-alkyl, C 2-4 alkoxy-C 1-4 alkyl, C 2-4 alkoxy-deuterated C 1-4 alkyl, oxo, -CONH2, C 3-6 alkenyl-C(O)NH-, C 2-4 alkynyl-NH-, -C(O)O-C 1-4 alkyl, -C(O)NH-C 1-4 alkyl,

[0042] In some embodiments, selected from

[0043] In some embodiments, the compound of formula (I), or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, is a compound of formula (Ia)-(Ig):

[0044]

[0045] wherein,

[0046] X1 is selected from CH, N, O or S;

[0047] Y2 is selected from CH, CR y , N, O or S;

[0048] M1 is selected from CH, N, O or S;

[0049] M4 is selected from CH, N, O or S;

[0050] R2 and R3 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6Hydroxyalkyl, oxo group, -CONH2;

[0051] Alternatively, R2 and R3 together with the C atom to which they are attached form a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S;

[0052] R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 2-6 cyano-alkyl, oxo group, -CONH2, C 3-6 alkenyl-C(O)NH-, C 2-6 alkynyl-NH-, -C(O)O-C 1-6 alkyl, -C(O)NH-C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 alkyl-O-C 1-6 alkyl, -C 1-6 alkyl-O-C 3-6 cycloalkyl, C 1-6 alkyl-O-C 1-6 haloalkyl, deuterated C 1-6 alkyl-O-C 1-6 haloalkyl, C 1-6 alkyl-a 4- to 6-membered heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 1-6 alkyl-C 6-10 cycloalkyl, C 1-6 alkyl-C 6-10 aryl, C 1-6 alkyl-a 5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O, a 4- to 6-membered saturated or partially unsaturated heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 6-10 aryl, a 5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O, wherein each of the alkyl, haloalkyl, hydroxyalkyl, cycloalkyl, heterocyclic group, oxo group, aryl, cycloalkyl, heterocyclic group and heteroaryl is optionally substituted, in each occurrence, with one or more substituents selected from halogen, amino, nitro, cyano, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 6-10Substituted with an aryl group and a substituent of a 5- to 7-membered heteroaryl group containing 1 or 2 heteroatoms selected from N or O;

[0053] Preferably, R5 is independently selected from i) hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 2-4 cyano-alkyl, oxo, -CONH2, C 3-6 alkenyl-C(O)NH-, C 2-4 alkynyl-NH-, -C(O)O-C 1-4 alkyl, -C(O)NH-C 1-4 alkyl, C 1-3 alkyl-O-C 1-5 alkyl, deuterated C 1-3 alkyl-O-C 1-3 haloalkyl, -C 1-3 alkyl-O-C 3-6 cycloalkyl, C 1-3 alkyl-O-C 1-6 haloalkyl, C 1-3 alkyl-O-N(C 1-3 alkyl)C(O)-, C 1-3 alkyl-N(C 1-3 alkyl)2, C 1-3 hydroxyalkoxy-C 1-3 alkyl, ii) Optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 2-4 cyano-alkyl, oxo, -CONH2, -C(O)O-C 1-4 alkyl, -C(O)NH-C 1-4 alkyl, C 1-3 alkyl-O-C 1-5 alkyl, deuterated C 1-3 alkyl-O-C 1-3 haloalkyl, C 1-3 alkyl-O-C 1-6 substituted with a substituent of haloalkyl;

[0054] R6 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl;

[0055] Preferably, R6 is independently selected from hydrogen, deuterium, F, Cl, Br, amino, cyano, nitro, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy and C 1-6 hydroxyalkyl;

[0056] R 2g 、R 3g 、R 4g 、R 5g are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, deuterated C 1-6 alkyl, deuterated C 2-6 alkenyl, deuterated C 2-6 alkynyl, deuterated C 1-6 haloalkyl, deuterated C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo, -CONH2;

[0057] R y is selected from hydrogen, deuterium, F, Cl, Br, amino, cyano, nitro, deuterated C 1-6 alkyl, deuterated C 2-6 alkenyl, deuterated C 2-6 alkynyl, deuterated C 1-6 haloalkyl, deuterated C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo and -CONH2;

[0058] t is 0, 1, 2 or 3.

[0059] In a preferred embodiment, R2 and R3 are each independently selected from hydrogen, deuterium, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl;

[0060] In a preferred embodiment, R2 and R3 together with the C atom to which they are attached form

[0061] In some embodiments, the compound of formula (I), or a tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereoisomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, is a compound of formula (IIa) or (IIb):

[0062]

[0063] wherein,

[0064] L is NH, O, S, C 1-3 alkylene, -C 1-3 alkyl-O-, -C 1-3 alkyl-NH- or -CONH-;

[0065] L1 is O, S or NH;

[0066] Y2 is selected from CH, N, O or S;

[0067] M4 is selected from CH, N, O or S;

[0068] R2 and R3 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo and -CONH2;

[0069] Alternatively, R2 and R3 together with the C atom to which they are attached form a C 3-6 cycloalkyl or a 4-6 membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S;

[0070] R4 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo and -CONH2;

[0071] R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 6-10 aryl, and a 5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O;

[0072] each R7 is independently selected from cyano, hydroxy, C 1-6 alkyl, deuterated C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, -NR a R b 、=NR b 、-C(O)NR a R b 、-NR a C(O)R b 、-C(O)OR b 、C 3-6 cycloalkyl, C 6-10 aryl, a 4- to 10-membered heterocyclic group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, a 5- to 10-membered heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S; wherein R7 is optionally substituted by one or more substituents selected from deuterium, halogen, amino, nitro, oxo, cyano, hydroxy, carboxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 1-6 alkoxy-C 1-6 alkyl, C 1-6 alkoxy-deuterated C 1-6 alkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S, a 5- to 6-membered heteroaryl containing 1 or 2 heteroatoms selected from N, O or S;

[0073] Alternatively, R7 and together form a 9- to 10-membered heterocyclic group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, a 9- to 10-membered heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S;

[0074] R a is independently selected from hydrogen, deuterium, C 1-6 alkyl, deuterated C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 hydroxyalkyl and C 3-6 cycloalkyl;

[0075] R b independently selected from hydrogen, deuterium, C 1-6 alkyl, deuterated C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S, and a 5- to 6-membered heteroaryl group containing 1 or 2 heteroatoms selected from N, O or S;

[0076] m is 0, 1 or 2;

[0077] n is 0, 1 or 2;

[0078] t is 0, 1 or 2; and

[0079] s is 0, 1, 2 or 3.

[0080] In some embodiments,

[0081] L is NH, O, S,

[0082] L1 is O, S or NH;

[0083] Y2 is selected from CH, N, O or S;

[0084] M4 is selected from CH, N, O or S;

[0085] R2 and R3 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy and C 1-3 hydroxyalkyl;

[0086] Alternatively, R2 and R3 together with the C atom to which they are attached form a C 3-6 cycloalkyl or a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S;

[0087] R4 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, oxo and -CONH2;

[0088] R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C1-3 Hydroxyalkyl and C 3-6 Cycloalkyl;

[0089] Each R7 is independently selected from cyano, hydroxy, C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 1-3 Hydroxyalkyl, -NR a R b , =NR b , -C(O)NR a R b , -NR a C(O)R b , -C(O)OR b , C 3-6 Cycloalkyl, phenyl, naphthyl, a 4- to 6-membered heterocyclic group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, a 5- to 6-membered heteroaryl group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S; wherein R7 is optionally substituted by one or more substituents selected from deuterium, halogen, amino, nitro, oxo, cyano, hydroxy, carboxyl, C 1-3 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-3 Halogenated alkyl, C 1-3 Alkoxy, C 1-3 Hydroxyalkyl, C 1-3 Alkoxy-C 1-3 Alkyl, C 1-3 Alkoxy-deuterated C 1-3 Alkyl, C 3-6 Cycloalkyl, a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S and a 5- to 6-membered heteroaryl group containing 1 or 2 heteroatoms selected from N, O or S;

[0090] R a Is independently selected from hydrogen, deuterium, C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 1-3 Hydroxyalkyl and C 3-6 Cycloalkyl;

[0091] R b Is independently selected from hydrogen, deuterium, C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Halogenated alkyl, C 1-3 Hydroxyalkyl, C 3-6Cycloalkyl, 4-6 membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S, and 5-6 membered heteroaryl group containing 1 or 2 heteroatoms selected from N, O or S.

[0092] The present invention also provides a method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof, which comprises the step of reacting a compound of formula (Xa) with a compound of formula (Xb),

[0093]

[0094] wherein,

[0095] X is OH, SH or NH2;

[0096] Hal is halogen;

[0097] R1, R2, R3, R4, R5, X1, X2, X3, X4, Y1, Y2, Y3, Y4, Y5, Y6, s and t are as defined in formula (I).

[0098] In some embodiments, in the above pharmaceutical composition, the amount of the compound, its tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereomer or a mixture thereof, or its deuterated derivative, or its pharmaceutically acceptable salt is about 0.1-95% by weight of the free base; preferably about 5-70%, such as 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%.

[0099] In some embodiments, the above pharmaceutical composition is formulated into tablets, capsules, liquid form or injection form.

[0100] In some embodiments, in the above pharmaceutical composition, the amount of the compound, its tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereomer or a mixture thereof, or its deuterated derivative, or its pharmaceutically acceptable salt is about 1-1000 mg; 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.

[0101] In some embodiments, a compound, its tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereomer or mixture thereof, or its deuterated derivative, 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 adapted accordingly.

[0102] In some embodiments, a compound, its tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereomer or mixture thereof, or its pharmaceutically acceptable salt is formulated in solid or liquid form, such as syrup, suspension, emulsion, tablet, capsule, powder, granule or lozenge.

[0103] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of any compound of formula (I)-(IV), or its tautomer, or its deuterated derivative, or its pharmaceutically acceptable salt and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0104] On the other hand, the present invention relates to a method for inhibiting MK2 kinase, which comprises administering to a subject in need an effective amount of any compound of formula (I)-(IV), or a pharmaceutical composition comprising the same.

[0105] On the other hand, the present invention provides a method for treating a disorder mediated by MK2 kinase or its mutant in a patient in need, which comprises the step of administering to the patient a compound or a pharmaceutically acceptable composition according to the present invention. These diseases include but are not limited to immune diseases, autoimmune diseases and inflammatory diseases, cardiovascular diseases, infectious diseases, bone resorption disorders, neurodegenerative diseases and proliferative diseases.

[0106] In some embodiments, the immune disease, autoimmune disease or inflammatory disease is selected from inflammatory bowel disease, ulcerative colitis, Crohn's disease, multiple sclerosis, psoriasis, arthritis, rheumatoid arthritis, osteoarthritis, juvenile arthritis, psoriatic arthritis, reactive arthritis, ankylosing spondylitis, cryopyrin-associated periodic syndromes, Muckle-Wells syndrome, familial cold autoinflammatory syndrome, neonatal-onset multisystem inflammatory disease, TNF receptor-associated periodic syndrome, acute and chronic pancreatitis, atherosclerosis, gout, ankylosing spondylitis, fibrotic diseases, liver fibrosis, idiopathic pulmonary fibrosis, kidney disease, sarcoidosis, scleroderma, allergic reactions, diabetes, type 1 diabetes, type 2 diabetes, diabetic retinopathy, Still's disease, vasculitis, sarcoidosis, pulmonary inflammation, acute respiratory distress syndrome, wet and dry age-related macular degeneration, autoimmune hemolytic syndrome, autoimmune and inflammatory hepatitis, autoimmune neuropathy, autoimmune ovarian failure, autoimmune orchitis, autoimmune thrombocytopenia, silicone implant-associated autoimmune disease, Sjögren's syndrome, familial Mediterranean fever, systemic lupus erythematosus, vasculitis syndromes, temporal, Takayasu's and giant cell arteritis, Beliefsdisease), Wegener's granulomatosis, vitiligo, secondary hematological manifestations of autoimmune diseases, anemia, drug-induced autoimmunity, Hashimoto's thyroiditis, hypophysitis, idiopathic thrombocytopenic purpura, metal-induced autoimmunity, myasthenia gravis, pemphigus, autoimmune deafness, Meniere's disease, Goodpasture's syndrome, Graves' disease, HW-related autoimmune syndrome, Guillain-Barre disease, Addison's disease, antiphospholipid syndrome, asthma, atopic dermatitis, celiac disease, Cushing's syndrome, dermatomyositis, idiopathic adrenal cortical atrophy, idiopathic thrombocytopenia, Kawasaki syndrome, Lambert-Eaton syndrome, pernicious anemia, hay fever, polyarteritis nodosa, primary biliary cirrhosis, primary sclerosing cholangitis, Raynaud's syndrome, Reiter's syndrome, relapsing polychondritis, Schmidt's syndrome, thyroiditis, sepsis, septic shock, endotoxic shock, exotoxin-induced toxic shock, gram-negative sepsis, toxic shock syndrome, glomerulonephritis, peritonitis, interstitial cystitis, hyperoxia-induced inflammation, chronic obstructive pulmonary disease (COPD), vasculitis, graft-versus-host reaction, graft-versus-host disease, allograft rejection, acute allograft rejection, chronic allograft rejection, early graft rejection, acute allograft rejection, reperfusion injury, pain, acute pain, chronic pain, neuropathic pain, fibromyalgia, chronic infection, meningitis, encephalitis, myocarditis, gingivitis, post-operative trauma, tissue injury, brain trauma, enterocolitis, sinusitis, uveitis, ocular inflammation, optic neuritis, gastric ulcer, esophagitis, peritonitis, periodontitis, dermatomyositis, gastritis, myositis, polymyalgia, pneumonia and bronchitis. Detailed implementation manners

[0107] The following are the definitions of the terms used in this application. Any term not defined herein has its normal meaning as understood by those skilled in the art.

[0108] "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 more 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 may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available attachment point. Preferably, the substituent is one or more substituents independently selected from alkyl, halogen, alkoxy, alkenyl, alkynyl, alkylsulfonyl, alkylamino, mercapto, hydroxy, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocyclic group, cycloalkylthio, heterocycloalkylthio and oxo group.

[0109] "Alkenyl" means an alkyl group as defined above having at least two carbon atoms and at least one carbon-carbon double bond, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc., preferably C 2-20Alkenyl, more preferably C 2-12 Alkenyl, most preferably C 2-6 Alkenyl. The alkenyl may be substituted or unsubstituted. When substituted, the substituents are preferably independently selected from one or more 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, sometimes preferably one to five groups, and sometimes more preferably one to three groups.

[0110] "Alkynyl" refers to an alkyl group as defined above having at least two carbon atoms and at least one carbon-carbon triple bond, such as ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, etc., preferably C 2-20 Alkynyl, more preferably C 2-12 Alkynyl, most preferably C 2-6 Alkynyl. The alkynyl may be substituted or unsubstituted. When substituted, the substituents are preferably independently selected from one or more of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio, sometimes preferably one to five groups, and sometimes more preferably one to three groups.

[0111] "Alkylene" refers to a saturated straight-chain or branched-chain aliphatic hydrocarbon group having 2 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-chain 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-ethylene (-CH(CH3)-), 1,2-ethylene (-CH2CH2-), 1,1-propylene (-CH(CH2CH3)-), 1,2-propylene (-CH2CH(CH3)-), 1,3-propylene (-CH2CH2CH2-), 1,4-butylene (-CH2CH2CH2CH2-), etc. The alkylene may be substituted or unsubstituted. When substituted, the substituents are preferably independently selected from one or more of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio, sometimes preferably one to five groups, and sometimes more preferably one to three groups.

[0112] "Alkenylene" refers to an alkylene group as defined above having at least two carbon atoms and at least one carbon-carbon double bond, preferably C 2-20 Alkenylene, more preferably C 2-12Alkenylene, 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. The alkenylene may be substituted or unsubstituted. When substituted, the substituents are preferably independently selected from one or more of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio, sometimes preferably one to five, and sometimes more preferably one to three groups.

[0113] "Alkynylene" refers to an alkynyl group as defined above having at least two carbon atoms and at least one carbon-carbon triple bond, preferably C 2-20 Alkynylene, more preferably C 2-12 Alkynylene, most preferably C 2-6 Alkynylene. Non-limiting examples of alkynylene include, but are not limited to, -CH≡CH-, -CH≡CHCH2-, -CH≡CHCH2CH2-, -CH2CH≡CHCH2-, etc. The alkynylene may be substituted or unsubstituted. When substituted, the substituents are preferably independently selected from one or more of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio, sometimes preferably one to five, and sometimes more preferably one to three groups.

[0114] "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, 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, fused or bridged rings.

[0115] "Spirocycloalkyl" refers to a 5- to 20-membered polycyclic group in which the rings are connected via 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 none of the rings has a fully conjugated π-electron system. Preferably, the spirocycloalkyl is 6 to 14 members, more preferably 7 to 10 members. Depending on the number of common spiro atoms, the spirocycloalkyl is divided into monospirocycloalkyl, dispirocycloalkyl or polyspirocycloalkyl, preferably referring to monospirocycloalkyl or dispirocycloalkyl, 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:

[0116]

[0117] "Fused cycloalkyl" means a 5- to 20-membered polycyclic hydrocarbon group in which each ring in the system shares an adjacent pair of carbon atoms with another ring, and 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 fused cycloalkyl is 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, the fused cycloalkyl is classified into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl, preferably referring 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:

[0118]

[0119] "Bridged cycloalkyl" means a 5- to 20-membered polycyclic hydrocarbon group in which every two rings in the system share two non-adjacent carbon atoms, and the 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 members, more preferably 7 to 10 members. Depending on the number of constituent rings, the bridged cycloalkyl is classified into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl, preferably referring to bicyclic, tricyclic or tetracyclic bridged cycloalkyl, more preferably bicyclic or tricyclic bridged cycloalkyl. Representative examples of bridged cycloalkyl include, but are not limited to, the following substituents:

[0120]

[0121] The cycloalkyl may be fused to a ring of an aryl, heteroaryl or heterocycloalkyl, and the ring connected to the parent structure is a cycloalkyl. Representative examples include, but are not limited to, indanylacetic acid, tetralin, benzocycloheptyl, etc. The cycloalkyl is optionally substituted or unsubstituted. When substituted, the substituents are preferably independently selected from one or more 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, sometimes preferably one to five, sometimes more preferably one to three substituents.

[0122] "Heterocyclic group" means a 3- to 20-membered saturated and / or partially unsaturated monocyclic or polycyclic hydrocarbon group having one selected from N, O and S(O) mOne or more of (where m is 0, 1 or 2), sometimes preferably one to five, and sometimes more preferably one to three heteroatoms as ring atoms, provided that 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; and 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, sulfo-morpholinyl, homopiperazinyl, etc. Polycyclic heterocyclic groups include heterocyclic groups having spiro, fused or bridged rings.

[0123] “Spiroheterocyclic group” refers to a 5- to 20-membered polycyclic heterocyclic group in which the rings are connected via a common carbon atom (referred to as a spiro atom), where the rings have one or more heteroatoms selected from N, O and S(O) m One or more of (where m is 0, 1 or 2), sometimes preferably one to five, and sometimes more preferably one to three heteroatoms as ring atoms, the remaining ring atoms being 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 members, more preferably 7 to 10 members. Depending on the number of common spiro atoms, the spiroheterocyclic group is divided into monospiroheterocyclic groups, dispiroheterocyclic groups or polyspiroheterocyclic groups, preferably referring to monospiroheterocyclic groups or dispiroheterocyclic groups, more preferably 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered or 5-membered / 6-membered monospiroheterocyclic groups. Representative examples of spiroheterocyclic groups include, but are not limited to, the following substituents:

[0124]

[0125]

[0126] “Fused heterocyclic group” refers to a 5- to 20-membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of carbon atoms with the other rings, 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 the ring has one or more heteroatoms selected from N, O and S(O) p One or more of (where p is 0, 1 or 2), sometimes preferably one to five, and sometimes more preferably one to three heteroatoms as ring atoms, the remaining ring atoms being C. Preferably, the fused heterocyclic group is 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, the fused heterocyclic group is divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups, preferably referring to bicyclic or tricyclic fused heterocyclic groups, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic groups. Representative examples of fused heterocyclic groups include, but are not limited to, the following substituents:

[0127]

[0128] "Bridged heterocyclic group" refers to 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 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, more preferably 7- to 10-membered. According to the number of constituent rings, the bridged heterocyclic group is classified into bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclic groups, preferably referring to bicyclic, tricyclic, or tetracyclic bridged heterocyclic groups, more preferably bicyclic or tricyclic bridged heterocyclic groups. Representative examples of the bridged heterocyclic group include, but are not limited to, the following substituents:

[0129]

[0130] The rings of the heterocyclic group may be fused to the rings of an aryl, heteroaryl, or cycloalkyl group, and the ring connected to the parent structure is the heterocyclic group. Representative examples include, but are not limited to, the following substituents:

[0131]

[0132] The heterocyclic group is optionally substituted or unsubstituted. When substituted, the substituents are preferably independently selected from one or more of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio, sometimes preferably one to five, and sometimes more preferably one to three groups.

[0133] "Aryl" refers to a 6- to 14-membered all-carbon monocyclic or polycyclic fused ring ("fused" ring system means that each ring in the system shares an adjacent pair of 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 to the rings of a heteroaryl, heterocyclic, or cycloalkyl group, and the ring connected to the parent structure is the aryl. Representative examples include, but are not limited to, the following substituents:

[0134]

[0135] The aryl may be substituted or unsubstituted. When substituted, the substituents are preferably independently selected from one or more of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio, sometimes preferably one to five, and sometimes more preferably one to three substituents.

[0136] "Heteroaryl" refers to an aryl system having 1 to 4 heteroatoms selected from 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 thiadiazolyl, pyrazolyl, oxazolyl, oxadiazolyl, imidazolyl, triazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl can be fused to a ring of an aryl, heterocyclic group, or cycloalkyl group, where the ring linked to the parent structure is the heteroaryl. Representative examples include, but are not limited to, the following substituents:

[0137]

[0138] The heteroaryl can be substituted or unsubstituted. When substituted, the substituents are preferably independently selected from one or more of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, and -NR 9 R 10 , sometimes preferably one to five, and sometimes more preferably one to three substituents.

[0139] "Alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), where the alkyl is as defined above. Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, etc. The alkoxy can be substituted or unsubstituted. When substituted, the substituents are preferably independently selected from one or more of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio, sometimes preferably one to five, and sometimes more preferably one to three substituents.

[0140] "Bond" refers to a covalent bond using the symbol "—".

[0141] "Hydroxyalkyl" refers to an alkyl substituted with a hydroxy group, where the alkyl is as defined above.

[0142] "Hydroxy" refers to the -OH group.

[0143] "Halogen" refers to a fluorine, chlorine, bromine, or iodine atom.

[0144] "Amino" refers to the -NH2 group.

[0145] "Cyano" refers to the -CN group.

[0146] "Nitro" refers to the -NO2 group.

[0147] "oxo group" means the =O group.

[0148] "carboxyl group" means the -C(O)OH group.

[0149] "carboxylate group" means -C(O)O(alkyl) or -C(O)O(cycloalkyl), where alkyl and cycloalkyl are as defined above.

[0150] "deuterated derivative" means a compound having the same chemical structure as a reference compound, but with one or more hydrogen atoms replaced by deuterium atoms ("D"). It should be recognized that some variation in natural isotope abundance will occur in the synthesized compound depending on the source of the chemical materials used in the synthesis. Despite this variation, the concentration of stable hydrogen isotopes at natural abundance is small and insignificant compared to the degree of stable isotope substitution of the deuterated derivatives described herein. Thus, unless otherwise stated, when referring to a "deuterated derivative" of a compound of the invention, at least one hydrogen is replaced by deuterium at a level far higher than its natural isotope abundance (which is typically about 0.015%). In some embodiments, the deuterated derivatives of the invention have an isotope enrichment factor of at least 3500 (52.5% deuterium incorporation at each designated deuterium), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), or at least 6600 (99% deuterium incorporation) for each deuterium atom.

[0151] "optional" or "optionally" means that the described event or circumstance may subsequently occur but does not have to occur, and the description includes instances where the event or circumstance may or may not occur. For example, "heterocyclyl optionally substituted with alkyl" means that the alkyl may be present but does not have to be present, and the description includes cases where the heterocyclyl is substituted with alkyl and cases where the heterocyclyl is not substituted with alkyl.

[0152] "substituted" means that one or more, preferably up to 5, more preferably 1 to 3, hydrogen atoms in a group are independently replaced by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions. A person skilled in the art can determine (by experiment or theory) what substitutions are possible or not without undue effort. For example, an amino or hydroxyl group with a free hydrogen may be unstable when bonded to a carbon atom with an unsaturated bond (such as an olefinic bond). For the purposes of the present invention, a heteroatom such as nitrogen can have a hydrogen substituent and / or any permitted substituent of the organic compounds described herein that satisfies the valence of the heteroatom.

[0153] "Pharmaceutical composition" refers to a mixture of one or more compounds according to the present invention, or their physiologically / pharmaceutically acceptable salts or prodrugs, and 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, facilitate the absorption of the active ingredient and thus exert its biological activity.

[0154] "Pharmaceutically acceptable salt" refers to a salt of a compound of the present invention that is safe and effective when used in mammals and has the corresponding biological activity.

[0155] Examples

[0156] The following examples are used to illustrate the present invention, but these examples should not be regarded as limiting the scope of the present invention. If the specific conditions of the experimental methods are not specified in the examples of the present invention, they are generally consistent with the conventional conditions or recommended conditions of the raw material and product manufacturers. Reagents without specific sources indicated are commercially available conventional reagents.

[0157] The structures of the compounds were identified by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The NMR chemical shift (δ) is given in 10 -6 (ppm). NMR was measured using a Varian Mercury 300 MHz Bruker Avance III 400 MHz instrument. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD).

[0158] High performance liquid chromatography (HPLC) was measured on an Agilent 1200 DAD high pressure liquid chromatograph (Sunfire C18 150×4.6 mm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18 150×4.6 mm column). Liquid chromatography-mass spectrometry (LCMS) was measured on an Agilent 1200 high pressure liquid chromatograph and mass spectrometer (Sunfire C18 4.6*50 mm 3.5 μm column) and an Agilent 19091S-433HP-5 high pressure liquid chromatograph and mass spectrometer (XBridge C18 4.6x50 mm 3.5 μm column).

[0159] Chiral high performance liquid chromatography (HPLC) was measured on an SFC Thar 80&150&200 (waters.).

[0160] The average percentage of ATPase inhibition and the IC 50 value were measured by a Victor Nivo multimode microplate reader (PerkinElmer, USA).

[0161] The thin-layer silica gel plate for thin-layer chromatography is the Yantai Xinnuo silica gel plate. The specifications of the plate used in TLC are from 0.15 mm to 0.2 mm, and the specifications of the plate used in thin-layer chromatography for product purification are from 0.4 mm to 0.5 mm.

[0162] Column chromatography usually uses silica gel with 200 - 300 mesh from Qingdao Ocean as the carrier.

[0163] The known starting materials of the present invention can be prepared by conventional synthesis methods in the prior art or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Shaoyuan Chemical Technology Co., Ltd., Darui Chemicals Company, etc.

[0164] Unless otherwise specified in the examples, the following reactions are carried out under an argon or nitrogen atmosphere.

[0165] The term "argon atmosphere" or "nitrogen atmosphere" means that the reaction flask is equipped with a balloon containing 1 L of argon or nitrogen.

[0166] The term "hydrogen atmosphere" means that the reaction flask is equipped with a balloon containing 1 L of hydrogen.

[0167] MS is mass spectrometry, where (+) represents the positive mode, usually giving M + 1 (or M + H) absorption, where M = molecular weight.

[0168] Synthesis procedure

[0169] Synthesis of intermediates

[0170] Synthesis of INT - 003

[0171]

[0172] Step 1. 6-(4,4,5,5 - Tetramethyl - 1,3,2 - dioxaborolan - 2 - yl)-3,4 - dihydroisoquinolin - 1(2H)-one (INT - 003)

[0173] To a solution of 6-bromo-3,4-dihydroisoquinolin-1(2H)-one 1 (4 g, 17.69 mmol, 1.0 eq) in 1,4-dioxane (40 mL) was added 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-1,3,2-dioxaborolane 2 (6.74 g, 26.54 mmol, 1.5 eq) and Pd(dppf)Cl2·CH2Cl2 (1.43 g, 1.77 mmol, 0.1 eq). The reaction mixture was stirred at 95 °C for 2 h. The reaction mixture was concentrated and the residue was purified by silica gel column chromatography (0 - 60% EtOAc in petroleum ether) to give the title product (4.8 g, 99% yield). LCMS-ESI (m / z): [M+H]+ C15H20BNO3 calcd 273.15; found 273.35.

[0174] Synthesis of Int-005

[0175]

[0176] Step 1: 2-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one: To a solution of 1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (250 mg, 1.84 mmol, 1.0 eq) in MTBE (20 mL) at room temperature was added 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-1,3,2-dioxaborolane (467 mg, 1.84 mmol, 1.1 eq), bis(1,5-cyclooctadiene)di-μ-methoxydiruthenium(I) (37 mg, 0.055 mmol, 0.03 eq) and 4,4'-di-tert-butyl-2,2'-bipyridine (30 mg, 0.11 mmol, 0.06 eq). The reaction mixture was stirred at 55 °C for 2 h. The reaction mixture was concentrated and the residue was purified by silica gel column using DCM and MeOH (20:1) as eluent to give 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one Int-005 (450 mg, 83%). LCMS-ESI (m / z): [M+H] + C 13 H 19 BN2O3, calcd 262.12; found 263.10.

[0177] Synthesis of Int-006

[0178]

[0179] Step 1:

[0180] 4-((4-Bromopyridin-2-yl)oxy)-2-chloro-5-iodopyrimidine Int-006: To a mixture of 2,4-dichloro-5-iodopyrimidine (3.2 g, 11.68 mmol, 1.0 equiv) and 4-bromopyridin-2-ol (2 g, 11.68 mmol, 1.0 equiv) in DMF (100 mL) was added NaHCO3 (1.96 g, 23.36 mmol, 2.0 equiv). The reaction was stirred at 100 °C for 2 h. The reaction was cooled to room temperature, the mixture was diluted with water (100 mL) and extracted with EtOAc (100 mL × 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4 and concentrated. The crude product was purified by silica gel column chromatography, eluting with PE and EtOAc (95:5 to 90:10), to give 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-iodopyrimidine Int-006 (2.1 g, 5.11 mol, 44%). LCMS-ESI (m / z): [M+H] + Calculated for C9H4BrClIN3O 411.83; found 411.99.

[0181] Synthesis of Int-007

[0182]

[0183] Step 1

[0184] 2-Bromo-3-fluoro-N-methoxy-N-methylisonicotinamide: To a solution of 2-bromo-3-fluoropyridine-4-carboxylic acid (3.00 g, 13.6 mmol) in DCM (15 mL) was added HATU (7.76 g, 20.4 mmol), methoxy(methyl)amine (1.25 g, 20.4 mmol) and DIEA (7.03 g, 5.4 mmol). The reaction was stirred at 25 °C under nitrogen for 6 h. The reaction mixture was concentrated in vacuo and then extracted with EtOAc (15 mL × 3). The combined organic layers were washed with brine (10 mL x3), dried over anhydrous Na2SO4. The residue was purified by flash column chromatography on silica gel (petroleum ether / ethyl acetate = 10:1) to give 2-bromo-3-fluoro-N-methoxy-N-methylisonicotinamide as a yellow oil (2.64 g, yield 74%). LCMS (m / z): [M+H] + Calculated for C8H9BrFN2O2 262.9; found 263.0.

[0185] Step 2

[0186] 1-(2-Bromo-3-fluoropyridin-4-yl)ethan-1-one: To a solution of 2-bromo-3-fluoro-N-methoxy-N-methylisonicotinamide (2.64 g, 10 mmol) in THF (15 mL) at 0 °C under nitrogen was added methylmagnesium bromide (3 M in Et2O, 10 mL, 30 mmol). The resulting mixture was warmed to 25 °C and stirred for 3 h. The reaction mixture was quenched with saturated NH4Cl solution and then extracted with EtOAc (15 mL x 3). The combined organic layers were washed with brine (10 mL x 3), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by flash column chromatography on silica gel (petroleum ether / ethyl acetate = 5:1) to give 1-(2-bromo-3-fluoropyridin-4-yl)ethan-1-one (1.8 g, yield 82%). LCMS (m / z): [M+H] + Calculated for C7H6BrFNO 217.9; found 217.9.

[0187] Step 3

[0188] 2-Bromo-1-(2-bromo-3-fluoropyridin-4-yl)ethan-1-one: To a solution of 1-(2-bromo-3-fluoropyridin-4-yl)ethan-1-one (1.80 g, 8.20 mmol) in HBr (30% in AcOH, 15 mL) was added Br2 (0.66 mL, 13.1 mmol). The reaction mixture was stirred at 25 °C under nitrogen for 16 h. The reaction mixture was quenched with methyl tert-butyl ether and then filtered. The filter cake was 2-bromo-1-(2-bromo-3-fluoropyridin-4-yl)ethan-1-one (2.5 g, crude). LCMS (m / z): [M+H] + Calculated for C7H5Br2FNO 295.8; found 295.9.

[0189] Step 4

[0190] 2-(2-Bromo-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one: At 0 °C under nitrogen, 2,4-piperidinedione (957 mg, 8.47 mmol), acetic acid (1.01 g, 16.9 mmol), and ammonium acetate (1.95 g, 25.4 mmol) were added to a solution of 2-bromo-1-(2-bromo-3-fluoropyridin-4-yl)ethan-1-one (2.50 g, crude) in EtOH (15 mL). The mixture was warmed to 80 °C and stirred for 16 h. The mixture was diluted with water (20 mL) and a precipitate formed. The precipitate was collected and lyophilized to give 2-(2-bromo-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (1.2 g, 45% yield). It was used in the next step without further purification. LCMS (m / z): [M+H] + C 12 H 10 Calculated for BrFN3O 309.99; found 310.0.

[0191] Step 5

[0192] 2-(3-Fluoro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one: At 100 °C under a nitrogen atmosphere, Pd2(dba)3 (149 mg, 0.16 mmol), t-BuXphos (136 mg, 0.32 mmol), and KOH (2.4 mL, 2 mol / L in water) were added to a solution of 2-(2-bromo-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (500 mg, 1.61 mmol) in 1,4-dioxane (19 mL). The mixture was microwaved for 1 h. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (5 mL × 3). The aqueous layer was lyophilized to give 2-(3-fluoro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (280 mg, 70% yield). LCMS (m / z): [M+H] + C 12 H 11 Calculated for FN3O2 248.0; found 248.0.

[0193] Synthesis of Int-008

[0194]

[0195] Step 1

[0196] 2-Chloro-4-(1-ethoxyvinyl)pyrimidine: A mixture of 2,4-dichloropyrimidine (2 g, 13.42 mmol, 1.0 equiv), tributyl(1-ethoxyvinyl)stannane (7.27 g, 20.14 mmol, 1.5 equiv) and Pd(PPh3)2Cl2 (471 mg, 0.67 mmol, 0.05 equiv) in DMF (20 mL) was stirred overnight at 80 °C under nitrogen. The mixture was diluted with EtOAc (40 mL), KF (10 g) was added to the mixture and stirred at room temperature for 1 h. The mixture was filtered through a Celite pad and washed with EtOAc (40 mL), then diluted with water (100 mL) and extracted with EtOAc (40 mL×3). The combined organic phases were washed with brine (50 mL×3), dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (20:1) to give 2-chloro-4-(1-ethoxyvinyl)pyrimidine (1.53 g, 63%). LCMS-ESI (m / z): [M+H] + Calculated for C8H9ClN2O 184.62; found 184.62.

[0197] Step 2

[0198] 2-Bromo-1-(2-chloropyrimidin-4-yl)ethan-1-one: NBS (1.62 g, 9.12 mmol, 1.1 equiv) was added to a solution of 2-chloro-4-(1-ethoxyvinyl)pyrimidine (1.53 g, 8.29 mmol, 1.0 equiv) in THF (27 mL) and water (3 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 h and then concentrated. The residue was diluted with water (50 mL) and extracted with EtOAc (50 mL×3). The combined organic phases were washed with saturated aqueous NaHCO3 (50 mL), dried over Na2SO4 and concentrated to give the crude product. The crude product was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (10:1) to give 2-bromo-1-(2-chloropyrimidin-4-yl)ethan-1-one (1.53 g, 78%). LCMS-ESI (m / z): [M+H] + Calculated for C6H4BrClN2O 235.47; found 235.47.

[0199] Step 3

[0200] 1-(tert-Butyl) 3-ethyl 3-(iodomethyl) piperidine-1,3-dicarboxylate: At -78 °C, LDA (38.5 mL, 2 M in THF, 76.94 mmol, 1.1 eq) was added dropwise to a solution of 1-(tert-butyl) 3-ethyl piperidine-1,3-dicarboxylate (18.0 g, 69.95 mmol, 1.0 eq) in THF (27 mL). The reaction mixture was stirred at -78 °C for 3 h. Then, methylene iodide (20.61 g, 76.94 mmol, 1.1 eq) was added dropwise to the reaction mixture. The reaction was stirred at -78 °C for another 1 h, then warmed to room temperature and stirred at room temperature for 2 days. The mixture was diluted with water (200 mL) and extracted with EtOAc (200 mL × 3). The combined organic phases were washed with brine (200 mL), dried over Na2SO4 and concentrated to give the crude product. The crude product was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (10:1), to give 1-(tert-butyl) 3-ethyl 3-(iodomethyl) piperidine-1,3-dicarboxylate (27.8 g, 93%). LCMS-ESI (m / z): [M+H] + C 14 H 24 INO4 Calculated 397.25; Found 397.25.

[0201] Step 4

[0202] 1-(tert-Butyl) 3-ethyl 3-((1,3-dioxoisoindolin-2-yl)methyl) piperidine-1,3-dicarboxylate: A mixture of 1-(tert-butyl) 3-ethyl 3-(iodomethyl) piperidine-1,3-dicarboxylate (27.8 g, 69.98 mmol, 1.0 eq), isatoic anhydride (11.3 g, 96.98 mmol, 1.1 eq) and K2CO3 (14.49 g, 104.97 mmol, 1.5 eq) in DMF (100 mL) was stirred under nitrogen at 100 °C overnight. The mixture was diluted with water (500 mL) and extracted with EtOAc (200 mL × 3). The combined organic phases were washed with brine (100 mL × 3), dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (1:1), to give 1-(tert-butyl) 3-ethyl 3-((1,3-dioxoisoindolin-2-yl)methyl) piperidine-1,3-dicarboxylate (21 g, 76%). LCMS-ESI (m / z): [M+H] + C 22 H 28 N2O6 Calculated 416.47; Found 416.47.

[0203] Step 5

[0204] 1-(tert-Butyl) 3-ethyl 3-(aminomethyl) piperidine-1,3-dicarboxylate: To a solution of 1-(tert-butyl) 3-ethyl 3-((1,3-dioxoisoindolin-2-yl)methyl) piperidine-1,3-dicarboxylate (21 g, 50.36 mmol, 1.0 equivalent) in MeOH (100 mL) was added hydrazine hydrate (2.77 g, 55.40 mmol, 1.1 equivalent). The reaction mixture was stirred at 65 °C for 5 h. The mixture was filtered and the filtrate was concentrated to give the crude product. The crude product was purified by silica gel column chromatography, eluting with DCM and MeOH (20:1) to give 1-(tert-butyl) 3-ethyl 3-(aminomethyl) piperidine-1,3-dicarboxylate (10.1 g, 70%). LCMS-ESI (m / z): [M+H] + C 14 H 26 Calculated for C14H27N3O4 286.37; found 286.77.

[0205] Step 6

[0206] 1-(tert-Butyl) 3-ethyl 3-(((2,4,6-trimethoxybenzyl)amino)methyl) piperidine-1,3-dicarboxylate: To a solution of 2,4,6-trimethoxybenzaldehyde (5.77 g, 29.39 mmol, 1.0 equivalent) and 1-(tert-butyl) 3-ethyl 3-(aminomethyl) piperidine-1,3-dicarboxylate (10.1 g, 35.27 mmol, 1.2 equivalent) in DCM (100 mL) was added acetic acid (0.1 mL), and the reaction mixture was stirred at room temperature for 30 min. Then, NaBH(OAc)3 (18.69 g, 88.17 mmol, 3 equivalents) was added to the reaction, and the mixture was stirred at room temperature for another 4 h. The reaction mixture was quenched with saturated aqueous NH4Cl solution (50 mL). The mixture was extracted with DCM and MeOH (15:1) (100 mL × 3). The combined organic phases were washed with brine (100 mL), dried over Na2SO4 and concentrated to give the crude product. The crude product was purified by silica gel column chromatography, eluting with DCM and MeOH (20:1) to give 1-(tert-butyl) 3-ethyl 3-(((2,4,6-trimethoxybenzyl)amino)methyl) piperidine-1,3-dicarboxylate (13.4 g, 98%). LCMS-ESI (m / z): [M+H] + C 24 H 38 Calculated for C22H37N3O7 466.58; found 466.58.

[0207] Step 7

[0208] 1-(tert-Butyl) 3-ethyl 3-((3-ethoxy-3-oxo-N-(2,4,6-trimethoxybenzyl) propanamido) methyl) piperidine-1,3-dicarboxylate: To a solution of 1-(tert-butyl) 3-ethyl 3-(((2,4,6-trimethoxybenzyl) amino) methyl) piperidine-1,3-dicarboxylate (13.4 g, 28.72 mmol, 1.0 equiv), DMAP (351 mg, 2.87 mmol, 0.1 equiv) and pyridine (6.82 g, 86.16 mmol, 3 equiv) in DCM (100 mL) was added methylmalonyl chloride (4.76 g, 31.59 mmol, 1.1 equiv), and the reaction was stirred at room temperature for 2 h. The reaction mixture was quenched with saturated aqueous NH4Cl (50 mL). The mixture was extracted with DCM and MeOH (15:1) (100 mL×3). The combined organic phases were washed with brine (100 mL), dried over Na2SO4 and concentrated to give the crude product. The crude product was purified by silica gel column chromatography, eluting with DCM and MeOH (20:1) to give 1-(tert-butyl) 3-ethyl 3-((3-ethoxy-3-oxo-N-(2,4,6-trimethoxybenzyl) propanamido) methyl) piperidine-1,3-dicarboxylate (15.0 g, 90%). LCMS-ESI (m / z): [M+H] + C 29 H 44 N2O 10 Calculated 580.68; Found 580.68.

[0209] Step 8

[0210] 2-(tert-Butyl) 10-ethyl 9,11-dioxo-8-(2,4,6-trimethoxybenzyl)-2,8-diazaspiro[5.5]undecane-2,10-dicarboxylate: Under nitrogen, to a solution of 1-(tert-butyl) 3-ethyl 3-((3-ethoxy-3-oxo-N-(2,4,6-trimethoxybenzyl) propanamido) methyl) piperidine-1,3-dicarboxylate (15.0 g, 25.83 mmol, 1.0 equiv) in MeOH (100 mL) was added sodium methoxide (9.30 g, 30% in MeOH, 51.66 mmol, 2.0 equiv), and the reaction was stirred at 65 °C for 3 h. The mixture was concentrated and purified by silica gel column chromatography, eluting with DCM and MeOH (20:1) to give 2-(tert-butyl) 10-ethyl 9,11-dioxo-8-(2,4,6-trimethoxybenzyl)-2,8-diazaspiro[5.5]undecane-2,10-dicarboxylate (10.3 g, 75%). LCMS-ESI (m / z): [M+H] + C 27 H 38N2O9 calculation: 534.61; measured: 534.61.

[0211] Step 9

[0212] tert-Butyl 9,11-dioxo-8-(2,4,6-trimethoxybenzyl)-2,8-diazaspiro[5.5]undecane-2-carboxylate Int-008: A mixture of 2-(tert-butyl) 10-ethyl 9,11-dioxo-8-(2,4,6-trimethoxybenzyl)-2,8-diazaspiro[5.5]undecane-2,10-dicarboxylate (10.3 g, 19.37 mmol, 1.0 equiv) in MeCN (30 mL) and water (30 mL) was stirred at 80 °C overnight. The mixture was concentrated and purified by silica gel column chromatography, eluting with DCM and MeOH (20:1) to afford tert-butyl 9,11-dioxo-8-(2,4,6-trimethoxybenzyl)-2,8-diazaspiro[5.5]undecane-2-carboxylate (8.5 g, 95%). LCMS-ESI (m / z): [M+H] + C 24 H 34 N2O7 calculation: 462.54; measured: 462.54.

[0213] Synthesis of Int-009

[0214]

[0215] tert-Butyl 2-(2-chloro-4-pyridyl)-4-oxo-5-[(2,4,6-trimethoxyphenyl)methyl]spiro[1,6-dihydropyrrolo[3,2-c]pyridine-7,3'-piperidine]-1'-carboxylate: A mixture of Int-008 (800 mg, 1.73 mmol, 1.0 equiv), 2-bromo-1-(2-chloro-4-pyridyl)ethenone (405 mg, 1.73 mmol, 1.0 equiv) and NH4OAc (532 mg, 6.92 mmol, 4 equiv) in EtOH (10 mL) was stirred at 60 °C overnight. The mixture was concentrated to give the crude product. The crude product was purified by silica gel column chromatography, eluting with DCM and MeOH (20:1) to afford tert-butyl 2-(2-chloro-4-pyridyl)-4-oxo-5-[(2,4,6-trimethoxyphenyl)methyl]spiro[1,6-dihydropyrrolo[3,2-c]pyridine-7,3'-piperidine]-1'-carboxylate (950 mg, 91%). LCMS-ESI (m / z): [M+H] + C 30 H 36 ClN5O6 calculation: 597.11; measured: 597.11.

[0216] Step 2

[0217] 2'-(2-chloropyridin-4-yl)-5',6'-dihydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-4'(1'H)-one: To a solution of tert-butyl 2-(2-chloro-4-pyridinyl)-4-oxo-5-[(2,4,6-trimethoxyphenyl)methyl]spiro[1,6-dihydropyrrolo[3,2-c]pyridine-7,3'-piperidine]-1'-carboxylate (950 mg, 1.59 mmol, 1.0 equiv) in DCM (10 mL) was added TFA (2 mL), and the reaction was stirred at room temperature for 16 h. The mixture was concentrated to afford crude 2'-(2-chloropyridin-4-yl)-5',6'-dihydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-4'(1'H)-one. LCMS-ESI (m / z): [M+H] + C 15 H 16 ClN5O calcd 316.79; found 316.79.

[0218] Step 3

[0219] tert-butyl 2-(2-chloro-4-pyridinyl)-4-oxo-spiro[5,6-dihydro-1H-pyrrolo[3,2-c]pyridine-7,3'-piperidine]-1'-carboxylate: To a solution of crude 2'-(2-chloropyridin-4-yl)-5',6'-dihydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-4'(1'H)-one in DCM (20 mL) were added TEA (2 mL), DMAP (46 mg, 0.38 mmol, 0.2 equiv) and (Boc)2O (1.65 g, 7.57 mmol, 2 equiv), and the reaction was stirred at room temperature overnight. The mixture was concentrated and purified by silica gel column chromatography, eluting with DCM and MeOH (20:1) to afford tert-butyl 2-(2-chloro-4-pyridinyl)-4-oxo-spiro[5,6-dihydro-1H-pyrrolo[3,2-c]pyridine-7,3'-piperidine]-1'-carboxylate (750 mg, 95%, 2 steps). LCMS-ESI (m / z): [M+H] + C 20 H 24 ClN5O3 calcd 416.91; found 416.91.

[0220] Step 4

[0221] tert-Butyl 2-(2-hydroxy-4-pyridinyl)-4-oxo-spiro[5,6-dihydro-1H-pyrrolo[3,2-c]pyridine-7,3'-piperidine]-1'-carboxylate Int-009: A mixture of tert-butyl 2-(2-chloro-4-pyridinyl)-4-oxo-spiro[5,6-dihydro-1H-pyrrolo[3,2-c]pyridine-7,3'-piperidine]-1'-carboxylate (600 mg, 1.44 mmol, 1.0 equiv), potassium hydroxide (242 mg, 4.32 mmol, 3.0 equiv), Pd2(dba)3 (131.79 mg, 0.14 mmol, 0.1 equiv), and t-Bu,bippyphos (145.82 mg, 0.29 mmol, 0.2 equiv) in 1,4-dioxane (20 mL) and water (5 mL) was stirred under nitrogen at 80 °C overnight. The mixture was concentrated and purified by silica gel column chromatography, eluting with DCM and MeOH (10:1) to give tert-butyl 2-(2-hydroxy-4-pyridinyl)-4-oxo-spiro[5,6-dihydro-1H-pyrrolo[3,2-c]pyridine-7,3'-piperidine]-1'-carboxylate (128 mg, 22%). LCMS-ESI (m / z): [M+H] + C 20 H 25 Calculated for C22H26N5O4 398.46; found 398.46.

[0222] Synthesis of Compound Int-0010

[0223]

[0224] Step 1

[0225] tert-Butyl 2'-(2-bromopyrimidin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate Int-0010: To a solution of Int-008 (8 g, 17.3 mmol) in EtOH (200 mL) at 25 °C was added 2-bromo-1-(2-bromopyridin-4-yl)ethan-1-one (5.8 g, 20.8 mmol), NH4OAc (8 g, 103.8 mmol), and AcOH (4.2 g, 69.2 mmol). The reaction mixture was heated to 80 °C and stirred at this temperature for 2 h under a nitrogen atmosphere with a balloon. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in EtOAc (200 mL) and washed with H2O (70 mL * 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (100% ethyl acetate) to give the product Int-006 (7 g, yield 63%). LCMS (m / z): [M+H]+ C 31 H 38 Calculated for CBrN4O6: 641.2; Found: 641.2.

[0226] Synthesis of Int-0011

[0227]

[0228] Step 1

[0229] tert-Butyl 2'-(2-hydroxypyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate: To a solution of Int-0010 (1 g, 1.56 mmol) in dioxane (9 mL) at room temperature under a nitrogen atmosphere with a balloon were added t-BuXphos (66 mg, 156 μmol), Pd2(dba)3 (142 mg, 156 μmol), H2O (1.56 mL), and KOH (349 mg, 6.24 mmol). The mixture was then heated to 100 °C for 2 h and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (methanol:dichloromethane = 1 / 15) to give tert-butyl 2'-(2-hydroxypyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (660 mg, yield 73%). LCMS (m / z): [M+H] + C 31 H 37 Calculated for CBrN4O6: 579.2; Found: 579.2.

[0230] Synthesis of Int-0012

[0231]

[0232] Step 1 2-(2-Chloropyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylic acid tert-butyl ester: A solution of 2-bromo-1-(2-chloro-4-pyridyl)ethanone 1 (1.65 g, 7.04 mmol, 1.0 equiv), tert-butyl 2,4-dioxopiperidine-1-carboxylate (1.5 g, 7.04 mmol, 1.0 equiv), and ammonium acetate (2.17 g, 28.16 mmol, 4.0 equiv) in EtOH (60 mL) was stirred at 60 °C overnight. After cooling to room temperature, the mixture was diluted with water (60 mL) and filtered. The solid was washed with water (20 mL × 2) and ether (20 mL), and then dried to give 2-(2-chloropyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylic acid tert-butyl ester (1.28 g, 3.68 mmol, 52%). LCMS-ESI (m / z): [M+H] + C 17 H 18 ClN3O3 Calcd 348.10; Found 347.99.

[0233] Step 2

[0234] 2-(2-Hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one Int-0012: A suspension of 2-(2-chloropyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylic acid tert-butyl ester (1 g, 2.88 mmol, 1.0 equiv) in HOAc (6 mL) and water (2 mL) was stirred in a sealed tube at 130 °C for 48 h. The reaction mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography, eluting with dichloromethane and methanol (7:1) to give Int-0012 (320 mg, 1.39 mmol, 48%). LCMS-ESI (m / z): [M+H] + C 12 H 11 N3O2 Calcd 230.09; Found 230.05.

[0235] Synthesis of Int-0013

[0236]

[0237] Step 1

[0238] 2-(2-((2-Chloro-5-iodopyrimidin-4-yl)oxy)pyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one Int-0013: At 25 °C under a nitrogen atmosphere, 2,4-dichloro-5-iodopyrimidine (1.17 g, 4.25 mmol) and NaHCO3 (714.63 mg, 8.5 mmol) were added to a solution of Int-0012 (650 mg, 2.83 mmol) in DMF (8 mL). Then the temperature was raised to 90 °C and stirred for 16 hours. The mixture was diluted with water (25 mL) and extracted with EA (25 mL × 3). The organic layer was washed with brine (20 mL) and concentrated under reduced pressure. The residue was purified by C18 reverse-phase column chromatography (H2O:ACN = 4 / 1) to give Int-0013 (400 mg, yield 27%). LCMS (m / z): [M+H]+ calculated 467.96; found 467.9.

[0239] Test method: Test method D was used.

[0240] Synthesis of Int-0014

[0241]

[0242] Step 1 2-Bromo-5-fluoro-N-methoxy-N-methylisonicotinamide: At 25 °C under a nitrogen atmosphere, HATU (12.96 g, 33.8 mmol), DIEA (11.75 g, 90.0 mmol) and N-methoxymethanamine (2.08 g, 33.8 mmol) were added to a solution of 2-bromo-5-fluoropyridine-4-carboxylic acid (5.00 g, 22.5 mmol) in dichloroethane (50 mL). The resulting mixture was stirred at 25 °C for 2 hours. The mixture was diluted with water (20 mL) and extracted with DCM (15 mL × 3). The organic phase was dried over Na2SO4, filtered and the filtrate was concentrated to give the crude product. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 10 / 1) to give 2-bromo-5-fluoro-N-methoxy-N-methylisonicotinamide (4.04 g, yield 66%). LCMS (m / z): [M+H]+ calculated 262.9; found 263.0.

[0243] Step 2

[0244] 1-(2-Bromo-5-fluoropyridin-4-yl)ethan-1-one: At 25 °C under a nitrogen atmosphere, methylmagnesium bromide (5.48 g, 45.9 mmol) was added to a solution of 2-bromo-5-fluoro-N-methoxy-N-methylisonicotinamide (4.04 g, 15.3 mmol) in THF (25 mL). The resulting mixture was stirred at 25 °C for 3 hours. The reaction mixture was quenched with saturated aqueous NH4Cl, diluted with ice water (10 mL) and extracted with EA (15 mL × 3). The combined organic phases were dried over Na2SO4, filtered and the filtrate was concentrated to give the crude product. The crude product was purified by flash column chromatography (petroleum ether: ethyl acetate = 5 / 1) to give 1-(2-bromo-5-fluoropyridin-4-yl)ethan-1-one (2.98 g, yield 65%). LCMS (m / z): [M+H]+ calculated 217.9; found 218.0.

[0245] Step 3

[0246] 2-Bromo-1-(2-bromo-5-fluoropyridin-4-yl)ethan-1-one: At 0 °C under a nitrogen atmosphere, Br2 (3.49 g, 21.28 mmol) was added to a solution of 1-(2-bromo-5-fluoropyridin-4-yl)ethan-1-one (2.98 g, 13.3 mmol) in HBr (15 mL, 0.3 mol / L in AcOH), then the temperature was raised to room temperature and stirred for 16 hours. The reaction mixture was transferred to a mixture of MTBE (15 mL) and filtered, and the filter cake was washed with EA to give the crude 2-bromo-1-(2-bromo-5-fluoropyridin-4-yl)ethan-1-one (3.1 g, yield 79%). It was used in the next step without further purification. LCMS (m / z): [M+H]+ calculated 297.8; found 297.9.

[0247] Step 4

[0248] 2-(2-Bromo-5-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one: At 0 °C under a nitrogen atmosphere, acetic acid (3.98 g, 53.3 mmol), ammonium acetate (6.16 g, 79.9 mmol), and 2,4-piperidinedione (3 g, 26.6 mmol) were added to a solution of 2-bromo-1-(2-bromo-5-fluoropyridin-4-yl)ethan-1-one (3.1 g, 13.3 mmol) in EtOH (30 mL). The mixture was warmed to 80 °C and stirred for 16 hours. Then it was diluted with water (15 mL) and extracted with EA (15 mL × 3). The combined organic phases were dried over Na2SO4, filtered, and the filtrate was concentrated to give the crude product. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 2 / 1) to give 2-(2-bromo-5-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (2.54 g, yield 46%). LCMS (m / z): [M+H]+ calculated 309.9; found 310.0.

[0249] Step 5

[0250] 2-(5-Fluoro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one Int-0014: At 100 °C under a nitrogen atmosphere, Pd2(dba)3 (85 mg, 0.29 mmol), t-BuXPhos (127 mg, 0.29 mmol), and KOH (2.5 mL, 2 mol / L in water) were added to a solution of 2-(2-bromo-5-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (500 mg, 1.59 mmol) in 1,4-dioxane (10 mL). The mixture was microwaved for 1 hour. The mixture was concentrated in vacuo, and the residue was purified by a C18 reverse-phase column (acetonitrile:water = 7 / 1) to give Int-0014 (227 mg, yield 39%). LCMS (m / z): [M+H]+ calculated 248.1; found 248.1.

[0251] Synthesis of Int-0016

[0252]

[0253] Step 1: Int-007 (3 g, 12.14 mmol, 1 equiv), 2,4-dichloro-5-iodopyrimidine (2.65 g, 9.71 mmol, 0.8 equiv) and NaHCO3 (3.05 g, 36.42 mmol, 3 equiv) were stirred in DMF (25 mL) at 90 °C under nitrogen for 3 h. The reaction mixture was purified directly by preparative HPLC (ACN-H2O, 0.1% FA) to give Int-0016-A as a white solid (600 mg, 10.1%). LCMS (m / z): [M+H]+ calculated 485.96; found 486.0. 1 1H NMR (400 MHz, d6-DMSO) δ 12.08 (s, 1H), 9.10 (s, 1H), 8.19 (d, J = 5.3 Hz, 1H), 7.82 (t, J = 5.4 Hz, 1H), 7.23 (d, J = 7.0 Hz, 1H), 7.02 (d, J = 2.4 Hz, 1H), 3.43 (t, J = 7.1 Hz, 2H), 2.90 (t, J = 6.7 Hz, 2H). Int-0016-B as a white solid (300 mg, 5.1%). LCMS (m / z): [M+H]+ calculated 485.96; found 486.0.

[0254] Synthesis of Int-0017

[0255]

[0256] Step 1: To a solution of Int-0014 (380 mg, 1.54 mmol, 1.0 equiv) in DMF (10 mL) was added 2,4-dichloro-5-iodopyrimidine (634 mg, 2.31 mmol, 1.5 equiv) and NaHCO3 (387 mg, 4.61 mmol, 3.0 equiv). The reaction mixture was stirred at 90 °C for 3 h and then filtered. The filtrate was purified by a reverse-phase column (acetonitrile:water = 2 / 5) to give Int-0017 as a yellow solid (220 mg, yield 27%). It was mixed with some isomers. LCMS (m / z): [M+H]+ calculated 485.96; found 486.0. 1 1H NMR (400 MHz, d6-DMSO) δ 11.97 (s, 1H), 9.07 (s, 1H), 8.39 (d, J = 3.0 Hz, 1H), 7.68 (d, J = 5.5 Hz, 1H), 7.20 (s, 1H), 7.04 (t, J = 2.6 Hz, 1H), 3.42 (td, J = 6.8, 2.2 Hz, 2H), 2.87 (t, J = 6.8 Hz, 2H).

[0257] Test method A: Titank C183u 50*2.1mm; Mobile phase: A: 0.1% formic acid in water; B: acetonitrile. Separation method: 0 - 0.2 minutes: 5% B, 0.2 - 2.5 minutes: 5% B - 95% B, 2.5 - 2.6 minutes: 95% B - 5% B, 2.6 - 3.5 minutes: 5% B. Flow rate: 1 mL / min; Column temperature: 25 °C

[0258] Test method B: XBRIDGE C18 2.1*50mm, 3.5um; Mobile phase: H2O(0.05% TFA) - ACN(0.05% TFA) ACN from 0% to 60% in 7 minutes, 7 - 8 minutes, ACN from 60% to 100%. Flow rate: 0.8 mL / min, Column temperature: 30 °C

[0259] Test method C: CHIRALPAK IA 4.6*250mm, 5μm; Mobile phase: 40% acetonitrile, water(0.1% FA). Flow rate: 25 mL / min, Column temperature: 30 °C

[0260] Test method D: XBRIDGE C18 2.1*50mm, 3.5um; Mobile phase: H2O(0.05% TFA), ACN(0.05% TFA); 7 minutes, ACN 10 - 100%; Flow rate: 1.0 mL / min; Column temperature: 45 °C.

[0261] Test method E: Gemini 5u C19 150*21.2mm; Mobile phase: H2O(0.5% NH4OH), ACN; 10.5 minutes, ACN 2 - 95%; Flow rate: 20 mL / min; Column temperature: 20 °C.

[0262] Test method F: XBRIDGE C18 2.12*150mm, 5um; H2O(0.1% FA) - CAN; ACN from 2% to 95% in 16 minutes; Flow rate: 20.0 mL / min; Column temperature 45 °C.

[0263] Synthesis of Example 1

[0264]

[0265] Step 1: At room temperature, add 2,4-dichloro-5-(ethoxymethyl)pyrimidine 1-2 (710 mg, 2.59 mmol, 1.0 equiv) and NaHCO3 (565 mg, 5.17 mmol, 2.0 equiv) to a solution of 1-1 (580 mg, 2.59 mmol, 1.0 equiv) in DMF (10 mL). Stir the reaction mixture at 100 °C for 2 h. Concentrate the reaction mixture. Dilute the residue with water (50 mL) and extract with EtOAc (50 mL × 3). Wash the combined organic phases with brine (50 mL), dry over Na2SO4 and concentrate to obtain the crude product. Purify the crude product by silica gel column chromatography, eluting with petroleum ether and EtOAc (20:1) to obtain compound 1-3 (380 mg, 32%). LCMS-ESI (m / z): [M+H] + C 12 H 11 BrClN3O2 Calcd 344.59; Found 345.83.

[0266] Step 2: At room temperature, add 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-1,3,2-dioxaborolane 1-5 (467 mg, 1.84 mmol, 1.1 equiv), bis(1,5-cyclooctadiene)di-μ-methoxydiruthenium(I) (37 mg, 0.055 mmol, 0.03 equiv) and 4,4'-di-tert-butyl-2,2'-bipyridine (30 mg, 0.11 mmol, 0.06 equiv) to a solution of 1-4 (250 mg, 1.84 mmol, 1.0 equiv) in MTBE (20 mL). Stir the reaction mixture at 55 °C for 2 h. Concentrate the reaction mixture and purify the residue by silica gel column chromatography, eluting with DCM and MeOH (20:1) to obtain compound 1-6 (450 mg, 83%). LCMS-ESI (m / z): [M+H] + C 13 H 19 BN2O3 Calcd 262.12; Found 263.10.

[0267] Step 3: A mixture of Compound 1-3 (50 mg, 0.15 mmol, 1.0 equiv), Compound 1-6 (95 mg, 0.36 mmol, 2.5 equiv), Ad2nBuP G3 (11 mg, 14.51 μmol, 0.1 equiv) and CsOAc (56 mg, 290.20 μmol, 2.0 equiv) in 1,4-dioxane (0.5 mL) and water (0.05 mL) was stirred overnight at 80 °C under nitrogen. The mixture was diluted with water (10 mL) and extracted with DCM (15 mL×3). The combined organic phases were washed with brine (50 mL), dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give Example 1 (35 mg, 60%).

[0268] LCMS-ESI (m / z): [M+H] + C 19 H 18 Calculated for ClN5O3: 399.84; Found: 399.97.

[0269] 1 1H NMR (DMSO-d6): δ 12.00 (s, 1H), 8.71 (s, 1H), 8.27 (d, J = 2.7 Hz, 1H), 7.63 (dd, J = 2.1 Hz, 2.7 Hz, 1H), 7.51 (d, J = 0.6 Hz, 1H), 7.12 (d, J = 0.6 Hz, 1H), 7.10 (s, 1H), 4.60 (s, 2H), 3.61 - 3.58 (m, 2H), 3.43 - 3.40 (m, 2H), 2.85 (t, J = 3.6 Hz, 2H), 1.19 (t, J = 3.6 Hz, 3H).

[0270] Test method: Titank C18 3u 50*2.1 mm; Mobile phase: A: 0.1% formic acid in water; B: acetonitrile. Separation method: 0 - 0.2 min: 5% B, 0.2 - 2.5 min: 5% B - 95% B, 2.5 - 2.6 min: 95% B - 5% B, 2.6 - 3.5 min: 5% B

[0271] Time (min) A B 0-0.2 5％ 0.2-2.5 From 5% to 95% 2.5-2.6 From 95% to 5% 2.6-3.5 5％

[0272] Flow rate: 1 mL / min, Column temperature: 25 °C, Retention time = 2.17 min

[0273] The examples in Table 1 below were synthesized using a similar route as described in Example 1 above and appropriate starting materials.

[0274] Table 1

[0275]

[0276]

[0277]

[0278]

[0279]

[0280]

[0281]

[0282]

[0283]

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297]

[0298]

[0299]

[0300]

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309] Synthesis of Example 212

[0310]

[0311] Step 1: To a solution of 4-bromopyridin-2-ol (580 mg, 2.59 mmol, 1.0 equiv) in DMF (10 mL) at room temperature was added 2,4-dichloro-5-(ethoxymethyl)pyrimidine (710 mg, 2.59 mmol, 1.0 equiv) and NaHCO3 (565 mg, 5.17 mmol, 2.0 equiv). The reaction mixture was stirred at 100 °C for 2 h. The reaction mixture was concentrated. The residue was diluted with water (50 mL) and extracted with EtOAc (50 mL × 3). The combined organic phases were washed with brine (50 mL), dried over Na2SO4 and concentrated to give the crude product. The crude product was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (20:1) to give 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-(ethoxymethyl)pyrimidine (380 mg, 32%). LCMS-ESI (m / z): [M+H] + C 12 H 11 Calculated for C9H8BrClN3O2 344.59; found 345.83.

[0312] Step 2: Mixture of 4-Bromopyridin-2-ol: A solution of 4-bromopyridin-2-ol (50 mg, 0.15 mmol, 1.0 equiv), Int-005 (95 mg, 0.36 mmol, 2.5 equiv), Ad2nBuP G3 (11 mg, 14.51 μmol, 0.1 equiv) and CsOAc (56 mg, 290.20 μmol, 2.0 equiv) in 1,4-dioxane (0.5 mL) and water (0.05 mL) was stirred under nitrogen at 80 °C overnight. The mixture was diluted with water (10 mL) and extracted with DCM (15 mL×3). The combined organic phases were washed with brine (50 mL), dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give Example 212 (35 mg, 60%). LCMS-ESI (m / z): [M+H] + C 19 H 18 ClN5O3 Calcd 399.84; Found 399.97. 1 1H NMR (DMSO-d6): δ 12.00 (s, 1H), 8.71 (s, 1H), 8.27 (d, J = 2.7 Hz, 1H), 7.63 (dd, J = 2.1 Hz, 2.7 Hz, 1H), 7.51 (d, J = 0.6 Hz, 1H), 7.12 (d, J = 0.6 Hz, 1H), 7.10 (s, 1H), 4.60 (s, 2H), 3.61 - 3.58 (m, 2H), 3.43 - 3.40 (m, 2H), 2.85 (t, J = 3.6 Hz, 2H), 1.19 (t, J = 3.6 Hz, 3H).

[0313] Test method: Test method A used. Retention time = 2.17 min

[0314] Synthesis of Example 213, 214

[0315]

[0316] Step 1: A mixture of Example 212 (60 mg, 150.06 μmol, 1.0 equiv), trimethylsilylacetylene (44 mg, 450.19 μmol, 3.0 equiv), Pd(PPh3)4 (35 mg, 30.01 μmol, 0.2 equiv), CuI (11 mg, 60.03 μmol, 0.4 equiv) and TEA (46 mg, 450.19 mmol, 3.0 equiv) in DCM (2 mL) was stirred under nitrogen at 50 °C overnight. The mixture was purified by silica gel column chromatography, eluting with DCM and MeOH (10:1) to give Example 213 as a yellow solid (100 mg, 140%). LCMS-ESI (m / z): [M+H]+ C 24 H 27 Calculated for C14H21N5O3Si: 461.60; Found: 462.04. 1 1H NMR (DMSO-d6): δ 11.98 (s, 1H), 8.73 (s, 1H), 8.25 (d, J = 5.4 Hz, 1H), 7.61 (d, J = 6.6 Hz, 1H), 7.47 (s, 1H), 7.12 (d, J = 2.4 Hz, 1H), 7.10 (s, 1H), 4.60 (s, 2H), 3.61 - 3.58 (m, 2H), 3.43 - 3.42 (m, 2H), 2.85 (t, J = 6.6 Hz, 2H), 1.18 (t, J = 6.9 Hz, 3H), 0.21 (t, J = 3.6 Hz, 9H).

[0317] Step 2: A mixture of Example 212 (100 mg, 316.64 μmol, 1.0 equiv), K2CO3 (59 mg, 433.29 μmol, 2.0 equiv) in MeOH (2 mL) was stirred at room temperature under air until Example 213 was completely consumed. The mixture was purified by preparative HPLC to give Example 214 (8 mg, 10%). LCMS-ESI (m / z): [M + H]+ + C 21 H 19 Calculated for C12H19N5O3: 389.42; Found: 390.04. 1 1H NMR (DMSO-d6): δ 11.98 (s, 1H), 8.74 (s, 1H), 8.26 (d, J = 4.8 Hz, 1H), 7.61 (dd, J = 5.4, 1.2 Hz, 1H), 7.47 (d, J = 1.2 Hz, 1H), 7.11 (d, J = 2.4 Hz, 1H), 7.09 (s, 1H), 4.61 (s, 2H), 4.35 (s, 1H), 3.61 - 3.58 (m, 2H), 3.43 - 3.40 (m, 2H), 2.85 (t, J = 6.9 Hz, 2H), 1.19 (t, J = 6.9 Hz, 3H).

[0318] Test method: Test method A was used; retention time = 2.05 minutes.

[0319] Synthesis of Example 215

[0320]

[0321] Step 1: Di-tert-butyl 2-(2-((2-chloro-5-(ethoxymethyl)pyrimidin-4-yl)oxy)pyridin-4-yl)-4-oxo-6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-1,5(4H)-dicarboxylate: A mixture of Example 212 (150 mg, 0.38 mmol, 1.0 equiv), di-tert-butyl dicarbonate (410 mg, 1.88 mmol, 5.0 equiv), DMAP (9.0 mg, 75 μmol, 0.2 equiv) and TEA (230 mg, 2.25 mmol, 6.0 equiv) in acetonitrile (5 mL) was stirred overnight at room temperature. The mixture was concentrated and purified by silica gel column chromatography to give di-tert-butyl 2-(2-((2-chloro-5-(ethoxymethyl)pyrimidin-4-yl)oxy)pyridin-4-yl)-4-oxo-6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-1,5(4H)-dicarboxylate (352 mg, crude). LCMS-ESI (m / z): [M+H] + C 29 H 34 ClN5O7 Calcd 599.21; Found 600.01.

[0322] Step 2: tert-Butyl 2-(2-hydroxypyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate: A mixture of di-tert-butyl 2-(2-((2-chloro-5-(ethoxymethyl)pyrimidin-4-yl)oxy)pyridin-4-yl)-4-oxo-6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-1,5(4H)-dicarboxylate (547 mg, 0.91 mmol, 1.0 equiv) in MeOH solution of NH3 (10 mL, 7M) was stirred overnight at room temperature. The mixture was concentrated and purified by silica gel column chromatography to give tert-Butyl 2-(2-hydroxypyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (140 mg, 47%). LCMS-ESI (m / z): [M+H] + C 17 H 19 N3O4 Calcd 329.14; Found 330.05.

[0323] Step 3: tert-Butyl 2-(2-((5-carbamoyl-2-chloropyrimidin-4-yl)oxy)pyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate: A mixture of tert-butyl 2-(2-hydroxypyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (70 mg, 0.21 mmol, 1.0 equiv), NaHCO3 (36 mg, 0.42 mmol, 2.0 equiv) and 2,4-dichloropyrimidine-5-carboxamide (82 mg, 0.42 mmol, 2.0 equiv) in DMF (1.5 mL) was stirred at 85 °C for 4 h. The mixture was concentrated and purified by silica gel column chromatography to give tert-butyl 2-(2-((5-carbamoyl-2-chloropyrimidin-4-yl)oxy)pyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (32 mg, 31%). LCMS-ESI (m / z): [M+H] + C 22 H 21 Calculated for C22H23ClN6O5 484.13; found 485.26.

[0324] Step 4: A mixture of tert-butyl 2-(2-((5-carbamoyl-2-chloropyrimidin-4-yl)oxy)pyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (32 mg, 0.07 mmol, 1.0 equiv) and 1N HCl (0.2 mL) in acetonitrile (2.0 mL) was stirred at room temperature for 4 h. The mixture was purified by preparative HPLC to give Example 215 (6.7 mg, 26%). LCMS-ESI (m / z): [M+H] + C 17 H 13 Calculated for C16H13ClN6O3 384.07; found 384.98. 1 1H NMR (DMSO-d6): δ 12.00 (s, 1H), 8.94 (s, 1H), 8.28 (d, J = 2.7 Hz, 1H), 7.97 - 7.95 (m, 2H), 7.64 (d, J = 2.7 Hz, 1H), 7.54 (s, 1H), 7.10 - 7.08 (m, 2H), 3.43 - 3.41 (m, 2H), 2.87 - 2.84 (m, 2H).

[0325] Test method: Test method A used; retention time = 1.84 min.

[0326] Synthesis of Example 216

[0327]

[0328] Step 1: To a mixture of 4,6-dichloropyrimidine (745 mg, 5.0 mmol, 1.0 equiv) and 4-bromopyridin-2-ol (870 mg, 5.0 mmol, 1.0 equiv) in DMF (40 mL) was added NaHCO3 (840 mg, 10.0 mmol, 2.0 equiv). The reaction was stirred at 100 °C for 2 h. The reaction was cooled to room temperature, the mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and concentrated. The crude product was purified by silica gel column chromatography, eluting with DCM and methanol (95:5 to 90:10), to give 4-((4-bromopyridin-2-yl)oxy)-6-chloropyrimidine (100 mg, 349.03 μmol, 7%). LCMS-ESI (m / z): [M+H] + Calculated for C9H5BrClN3O 284.93; found 285.07.

[0329] Step 2: To a solution of 4-((4-bromopyridin-2-yl)oxy)-6-chloropyrimidine (40 mg, 139.61 μmol, 1.0 equiv) and 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (91 mg, 349.03 μmol, 2.5 equiv) in 1,4-dioxane (6 mL) and water (0.6 mL) was added Ad2nBu-G3 (10 mg, 13.96 μmol, 0.1 equiv) and cesium acetate (53 mg, 279.22 μmol, 2.0 equiv). The reaction was stirred at 80 °C overnight. The mixture was filtered and the solution was concentrated. The residue was purified by silica gel column chromatography to give Example 216 (25 mg, 73.15 μmol, 52%). LCMS-ESI (m / z): [M+H] + C 16 H 12 Calculated for C16H16ClN5O2 341.07; found 341.94. 1 1H NMR (DMSO-d6): δ 11.97 (s, 1H), 8.75 (s, 1H), 8.27 (d, J = 5.4 Hz, 1H), 7.62 (dd, J = 1 Hz, 5.4 Hz, 1H), 7.56 (d, J = 0.6 Hz, 1H), 7.50 (d, J = 1.2 Hz, 1H), 7.12 (d, J = 2.4 Hz, 1H), 7.09 (s, 1H), 3.42 - 3.40 (m, 2H), 2.86 - 2.83 (m, 2H).

[0330] Test method: Test method A was used; retention time = 1.91 minutes

[0331] Synthesis of Example 217

[0332]

[0333] Step 1: A solution of 5-(trifluoromethyl)-1H-pyrimidine-2,4-dione (2.16 g, 12.0 mmol, 1.0 equiv) and acetylhydrazine (1.16 g, 15.6 mmol, 1.3 equiv) in 1 M NaOH (48 mL, 48.0 mmol, 4.0 equiv) was stirred at 80 °C for 48 h. After cooling the mixture to room temperature, 1 M HCl (50 mL) was added with stirring. The mixture was filtered and the cake was washed with 1 M HCl (50 mL) and water (50 mL × 4). The solid was dried to give 5-(5-methyl-1,3,4-oxadiazol-2-yl)pyrimidine-2,4(1H,3H)-dione (1.20 g, 6.18 mmol, 51%). LCMS-ESI (m / z): [M+H] + Calculated for C7H6N4O3 195.04; found 195.02

[0334] Step 2: DIEA (1.94 g, 15.00 mmol, 2.6 mL, 5.0 equiv) was added to a suspension of 5-(5-methyl-1,3,4-oxadiazol-2-yl)pyrimidine-2,4(1H,3H)-dione (583 mg, 3.00 mmol, 1.0 equiv) in POCl3 (12 mL) at room temperature. The reaction mixture was stirred at 100 °C overnight. The reaction was poured slowly into water (200 mL) and the mixture was extracted with EtOAc (120 mL × 2). The combined organic layers were washed with brine (150 mL × 2), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (3:1) to give 2-(2,4-dichloropyrimidin-5-yl)-5-methyl-1,3,4-oxadiazole (455 mg, 1.97 mmol, 65%). LCMS-ESI (m / z): [M+H] + Calculated for C7H4Cl2N4O 230.98; found 230.96, 232.97

[0335] Step 3: A solution of 2-bromo-1-(2-chloro-4-pyridyl)ethanone (1.65 g, 7.04 mmol, 1.0 equiv), tert-butyl 2,4-dioxopiperidine-1-carboxylate (1.50 g, 7.04 mmol, 1.0 equiv), and ammonium acetate (2.17 g, 28.16 mmol, 4.0 equiv) in EtOH (60 mL) was stirred at 60 °C overnight. The reaction was cooled to room temperature, the mixture was diluted with water (60 mL) and the mixture was filtered. The solid was washed with water (20 mL × 2) and diethyl ether (20 mL), dried to give tert-butyl 2-(2-chloropyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (1.28 g, 3.68 mmol, 52%). LCMS-ESI (m / z): [M+H] + C 17 H 18 ClN3O3 Calcd 348.10; Found 347.99.

[0336] Step 4: 2-(2-Hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one:

[0337] A suspension of tert-butyl 2-(2-chloropyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (1 g, 2.88 mmol, 1.0 equiv) in acetic acid (6 mL) and water (2 mL) was stirred in a sealed tube at 130 °C for 48 h. The reaction mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography, eluting with dichloromethane and methanol (7:1) to give 2-(2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (320 mg, 1.39 mmol, 48%). LCMS-ESI (m / z): [M+H] + C 12 H 11 N3O2 Calcd 230.09; Found 230.05.

[0338] Step 5: 2-(2-((2-Chloro-5-(5-methyl-1,3,4-oxadiazol-2-yl)pyrimidin-4-yl)oxy)pyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one Example 217: To a solution of 2-(2,4-dichloropyrimidin-5-yl)-5-methyl-1,3,4-oxadiazole (50 mg, 218 μmol, 1.0 equiv) in NMP (1 mL) was added sodium bicarbonate (55 mg, 654 μmol, 3.0 equiv), and the mixture was stirred at room temperature for 30 minutes. Then 2-(2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (60 mg, 262 μmol, 1.2 equiv) was added, and the reaction mixture was stirred at 80 °C for 2 hours. The reaction mixture was purified by preparative HPLC to give Example 217 (2.8 mg, 6.6 μmol, 3%). LCMS-ESI (m / z): [M+H] + C 19 H 14 ClN7O3 Calculated 424.08; Found 423.94, 426.03. 1 1H NMR (600 MHz, DMSO-d6): δ 12.02 (s, 1H), 9.27 (s, 1H), 8.30 (d, J = 5.4 Hz, 1H), 7.66 (d, J = 6.8 Hz, 1H), 7.59 (s, 1H), 7.13 - 7.11 (m, 2H), 3.42 (m, 2H), 2.85 (t, J = 6.8 Hz, 2H), 2.63 (s, 3H).

[0339] Test method:

[0340] Test method A was used.

[0341] Retention time = 1.96 minutes

[0342] Synthesis of Example 218

[0343]

[0344] Step 1: To a solution of Int-0013 (400 mg, 0.97 mmol, 1.0 eq) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (284 mg, 1.46 mmol, 1.5 eq) in 1,4-dioxane (20 mL) and water (4 mL) was added Pd(dppf)Cl2-DCM (79 mg, 97 μmol, 0.1 eq) and NaHCO3 (244 mg, 2.91 mmol, 3.0 eq). The reaction was stirred at 55 °C overnight. The mixture was filtered and the solution was concentrated. The residue was purified by silica gel column chromatography to give 4-(4-((4-bromopyridin-2-yl)oxy)-2-chloropyrimidin-5-yl)isoxazole (180 mg, 0.51 mol, 52%). LCMS-ESI (m / z): [M+H] + C 12 H6BrClN4O2 Calcd 352.94; Found 352.99.

[0345] Step 2: To a solution of 4-(4-((4-bromopyridin-2-yl)oxy)-2-chloropyrimidin-5-yl)isoxazole (100 mg, 0.28 mmol, 1.0 eq) and 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (147 mg, 0.56 mmol, 2.0 eq) in 1,4-dioxane (10 mL) and water (2 mL) was added Pd(dppf)Cl2-DCM (23 mg, 28 μmol, 0.1 eq) and K3PO4 (178 mg, 0.84 mmol, 3.0 eq). The reaction was stirred in a microwave at 65 °C for 1.5 h. The mixture was filtered and the solution was concentrated. The residue was purified by preparative HPLC to give Example 218 (7.5 mg, 18 μmol, 6%). LCMS-ESI (m / z): [M+H] + C 19 H 13 ClN6O3 Calcd 409.07; Found 408.90. 1 1H NMR (DMSO-d6): δ 11.99 (s, 1H), 9.60 (s, 1H), 9.34 (s, 1H), 9.18 (s, 1H), 8.30 (d, J = 5.4 Hz, 1H), 7.65 (m, 1H), 7.59 (s, 1H), 7.10 (d, J = 1.8 Hz, 1H), 7.07 (s, 1H), 3.42 - 3.40 (m, 2H), 2.86 - 2.83 (m, 2H).

[0346] Test method: Test method A was used; retention time = 10.8 minutes

[0347] Synthesis of Example 219

[0348]

[0349] Step 1: A mixture of Int-0013 (300 mg, 728 μmol, 1.0 equiv), 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (144 mg, 692 μmol, 0.95 equiv), Pd(dppf)Cl2.DCM (60 mg, 72.8 μmol, 0.1 equiv) and NaHCO3 (184 mg, 2.18 mmol, 3.0 equiv) in a mixture of 1,4-dioxane (7.5 mL) and water (1.5 mL) was stirred overnight at 50 °C under nitrogen. The mixture was diluted with water (10 mL) and extracted with EtOAc (15 mL × 3). The combined organic phases were washed with brine (10 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-(1-methyl-1H-pyrazol-3-yl)pyrimidine (3.8 mg, 14%). LCMS-ESI (m / z): [M+H] + C 13 H9BrClN5O Calculated 366.60; Found 366.79.

[0350] Step 2: A mixture of 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-(1-methyl-1H-pyrazol-3-yl)pyrimidine (33 mg, 90 μmol, 1.0 equiv), 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (59 mg, 224 μmol, 2.5 equiv), Ad2nBuP G3 (7 mg, 9 μmol, 0.1 equiv) and CsOAc (35 mg, 180 μmol, 2.0 equiv) in a mixture of 1,4-dioxane (0.5 mL) and water (0.05 mL) was stirred overnight at 80 °C under nitrogen. The mixture was diluted with water (5 mL) and extracted with DCM (5 mL × 3). The combined organic phases were washed with brine (10 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give Example 219 (15 mg, 40%). LCMS-ESI (m / z): [M+H] + C 20 H 16 ClN7O2 Calculated 421.85; Found 421.96.1 1H NMR (DMSO-d6): δ 11.96 (s, 1H), 9.14 (s, 1H), 8.27 (d, J = 5.4 Hz, 1H), 7.86 (d, J = 2.4 Hz, 1H), 7.63 (t, J = 4.8 Hz, 1H), 7.57 (s, 1H), 7.13 (d, J = 2.4 Hz, 1H), 7.07 (s, 1H), 6.80 (d, J = 2.4 Hz, 1H), 3.96 (s, 3H), 3.42 (d, J = 2.4 Hz, 2H), 2.85 (t, J = 7.2 Hz, 3H).

[0351] Test method: Test method A was used; retention time = 2.07 minutes.

[0352] Synthesis of Example 220

[0353]

[0354] Step 1: To a solution of 4-bromopyridin-2-ol (87 mg, 0.5 mmol, 1 equiv) in DMF (1 mL) was added 2,4-dichloro-5-phenylpyrimidine (135 mg, 0.6 mmol, 1.2 equiv) and sodium bicarbonate (84 mg, 1 mmol, 2 equiv). The mixture was stirred at 100 °C for 2 h. The reaction mixture was concentrated in vacuo and purified by silica gel column (EA / PE = 3% - 5%) to give 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-phenylpyrimidine (180 mg, 0.49 mmol, 99%). LCMS (m / z): [M+H] + Calculated for C15H9BrClN3O 361.96; found 362.08.

[0355] Step 2: To a solution of 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-phenylpyrimidine (80 mg, 0.22 mmol, 1 equiv) in dioxane (2 mL) and H2O (0.2 mL) was added Int-005 (144 mg, 0.55 mmol, 2.5 equiv), Ad2nBuPG3Pd (16 mg, 0.02 mmol), CsOAc (84 mg, 0.44 mmol, 2 equiv), and the reaction was stirred at 80 °C overnight. The reaction mixture was purified by preparative HPLC to give Example 220 as a yellow solid (5.4 mg, 0.012 mmol, 5%). LCMS (m / z): [M+H] + Calculated for C22H16ClN5O2 418.10; found 417.95. 11H NMR (600 MHz, DMSO) δ 11.96 (s, 1H), 8.85 (s, 1H), 8.26 (d, J = 5.4 Hz, 1H), 7.75 - 7.71 (m, 2H), 7.62 - 7.58 (m, 1H), 7.57 - 7.51 (m, 3H), 7.50 - 7.46 (m, 1H), 7.11 (d, J = 2.4 Hz, 1H), 7.07 - 7.05 (m, 1H), 3.41 (dt, J = 6.9, 2.4 Hz, 2H), 2.85 (t, J = 6.9 Hz, 2H)

[0356] Test method: Test method A was used; retention time = 2.39 minutes

[0357] Synthesis of Example 221

[0358]

[0359] Step 1: To a solution of Int-0013 (600 mg, 1.46 mmol, 1.0 equiv) and pyridin-3-ylboronic acid (269 mg, 2.19 mmol, 1.5 equiv) in 1,4-dioxane (30 mL) and water (6 mL) was added Pd(dppf)Cl2-DCM (119 mg, 146 μmol, 0.1 equiv) and NaHCO3 (368 mg, 4.38 mmol, 3.0 equiv). The reaction was stirred overnight at 55 °C. The mixture was filtered and the solution was concentrated. The residue was purified by silica gel column chromatography to give 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-(pyridin-3-yl)pyrimidine as a white solid (180 mg, 0.50 mmol, 23%). LCMS-ESI (m / z): [M+H] + C 14 Calculated for C18H8BrClN4O 362.96; found 362.89

[0360] Step 2: To a solution of 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-(pyridin-3-yl)pyrimidine (180 mg, 0.50 mmol, 1.0 equiv) and Int-005 (262 mg, 1 mmol, 2.0 equiv) in 1,4-dioxane (10 mL) and water (2 mL) was added Pd(dppf)Cl2-DCM (41 mg, 50 μmol, 0.1 equiv) and K3PO4 (318 mg, 1.5 mmol, 3.0 equiv). The reaction was stirred in a microwave at 65 °C for 1.5 h. The mixture was filtered and the solution was concentrated. The residue was purified by preparative HPLC to give Example 221 (22 mg, 53 μmol, 11%). LCMS-ESI (m / z): [M+H] + C21 H 15 Calculated for ClN6O2: 419.09; Found: 418.87. 1 1H NMR (DMSO-d6): δ 11.96 (s, 1H), 8.93 (s, 1H), 8.92 (d, J = 1.8 Hz, 1H), 8.67 - 8.66 (m, 1H), 8.26 (d, J = 5.4 Hz, 1H), 8.16 - 8.14 (m, 1H), 7.61 - 7.60 (dd, J = 1.8 Hz, 1H), 7.58 - 7.56 (m, 2H), 7.10 (d, J = 2.4 Hz, 1H), 7.06 (s, 1H), 3.42 - 3.39 (m, 2H), 2.85 - 2.83 (m, 2H).

[0361] Test method: Test method A was used; retention time = 1.947 minutes

[0362] Synthesis of Example 222

[0363]

[0364] Step 1: To a solution of Int-007 (100 mg, 0.404 mmol) in ACN (3 mL) was added 2,4-dichloro-5-(ethoxymethyl)pyrimidine (67 mg, 0.323 mmol) and K2CO3 (167 mg, 1.213 mmol). The mixture was stirred at 60 °C for 3 hours. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (5 mL × 3). The combined organic layers were concentrated in vacuo, and the residue was purified by preparative HPLC (CAN-H2O, 0.1% TFA) to give Example 222 (2.41 mg, 2%). LCMS (m / z): [M + H] + C 19 H 18 Calculated for ClFN5O3: 418.1; Found: 418.1. 1 1H NMR (DMSO-d6): δ 12.05 (s, 1H), 8.75 (s, 1H), 8.18 (d, J = 5.2 Hz, 1H), 7.80 (t, J = 5.2 Hz, 1H), 7.22 - 7.16 (m, 1H), 7.01 - 6.96 (m, 1H), 4.64 (s, 2H), 3.60 (q, J = 6.9 Hz, 2H), 3.46 - 3.38 (m, 2H), 2.88 (t, J = 6.8 Hz, 2H), 1.18 (t, J = 7.0 Hz, 3H).

[0365] Test method: Test method B was used.

[0366] Synthesis of Example 223

[0367]

[0368] Step 1: To a solution of 1,2-oxazol-4-ylboronic diol (500 mg, 4.43 mmol) in 1,4-dioxane / H2O (5 mL) was added 2,4-dichloro-5-iodopyrimidine (1461 mg, 5.32 mmol), potassium carbonate (1836 mg, 13.29 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (321 mg, 0.44 mmol). The reaction mixture was stirred at 50 °C under nitrogen for 6 h and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 3 / 1) to give product 3 (80 mg, yield 5%). LCMS (m / z): [M+H]+ C7H4Cl2N3O calcd 216.0; found 216.0.

[0369] Step 2: To a solution of 2,4-dichloro-5-(1,2-oxazol-4-yl)pyrimidine 3 (55 mg, 0.25 mmol, 1.0 equiv) in DMF (5 mL) was added 2-(3-fluoro-2-hydroxypyridin-4-yl)-1H,5H,6H,7H-pyrrolo[3,2-c]pyridin-4-one 4 (76 mg, 0.31 mmol) and NaHCO3 (64 mg, 0.76 mmol). The reaction mixture was stirred at 50 °C under nitrogen for 2 h. It was diluted with water and extracted with ethyl acetate (10 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by preparative HPLC on silica gel (acetonitrile:H2O (0.1% FA) = 30–70%) to give product HBP-124-BD-CI-063 (1.48 mg, yield 1.34%). LCMS (m / z): [M+H]+ C 19 H 13 ClFN6O3 calcd 427.1; found 427.0. 1 H NMR (400 MHz, d6-DMSO) δ 12.09 (s, 1H), 9.67 (s, 1H), 9.37 (s, 1H), 9.22 (s, 1H), 8.18 (d, J = 5.3 Hz, 1H), 7.83–7.80 (m, 1H), 7.16 (s, 1H), 6.95 (s, 1H), 3.40–3.36 (m, 2H), 2.87–2.83 (m, 2H).

[0370] Test method: Test method C used; retention time = 1.07 min

[0371] Synthesis of Example 224

[0372]

[0373] Step 1: At 25 °C under a nitrogen atmosphere, add HATU (12.96 g, 33.8 mmol), DIEA (11.75 g, 90.0 mmol), and N - methoxymethylamine (2.08 g, 33.8 mmol) to a solution of 2 - bromo - 5 - fluoropyridine - 4 - carboxylic acid (5.00 g, 22.5 mmol) in dichloroethane (50 mL). Stir the resulting mixture at 25 °C for 2 hours. Dilute the mixture with water (20 mL) and extract with DCM (15 mL × 3). Dry the organic phase over Na2SO4, filter, and concentrate the filtrate to obtain the crude product. Purify the crude product by flash column chromatography (petroleum ether:ethyl acetate = 10 / 1) to obtain 2 - bromo - 5 - fluoro - N - methoxy - N - methylisonicotinamide as a white solid (4.04 g, yield 66%). LCMS (m / z): [M + H] + Calculated for C8H9BrFN2O2 262.9; found 263.0.

[0374] Step 2: At 25 °C under a nitrogen atmosphere, add methylmagnesium bromide (5.48 g, 45.9 mmol) to a solution of 2 - bromo - 5 - fluoro - N - methoxy - N - methylisonicotinamide (4.04 g, 15.3 mmol) in THF (25 mL). Stir the resulting mixture at 25 °C for 3 hours. Quench the reaction mixture with saturated aqueous NH4Cl, dilute with ice water (10 mL), and extract with EA (15 mL × 3). Dry the combined organic phases over Na2SO4, filter, and concentrate the filtrate to obtain the crude product. Purify the crude product by flash column chromatography (petroleum ether:ethyl acetate = 5 / 1) to obtain 1 - (2 - bromo - 5 - fluoropyridin - 4 - yl)ethan - 1 - one (2.98 g, yield 65%). LCMS (m / z): [M + H] + Calculated for C7H6BrFNO 217.9; found 218.0.

[0375] Step 3: 2-Bromo-1-(2-bromo-5-fluoropyridin-4-yl)ethan-1-one: At 0 °C under a nitrogen atmosphere, Br2 (3.49 g, 21.28 mmol) was added to a solution of 1-(2-bromo-5-fluoropyridin-4-yl)ethan-1-one (2.98 g, 13.3 mmol) in HBr (15 mL, 0.3 mol / L in AcOH). Then the temperature was raised to room temperature and stirred for 16 h. The reaction mixture was transferred to a mixture of MTBE (15 mL). Then it was filtered, and the filter cake was washed with EA to obtain crude 2-bromo-1-(2-bromo-5-fluoropyridin-4-yl)ethan-1-one as a yellow solid (3.1 g, yield 79%). It can be used for the next step without further purification. LCMS (m / z): [M+H] + Calculated for C7H5Br2FNO 297.8; found 297.9.

[0376] Step 4: 2-(2-Bromo-5-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one: At 0 °C under a nitrogen atmosphere, acetic acid (3.98 g, 53.3 mmol), ammonium acetate (6.16 g, 79.9 mmol), and 2,4-piperidinedione (3 g, 26.6 mmol) were added to a solution of 2-bromo-1-(2-bromo-5-fluoropyridin-4-yl)ethan-1-one (3.1 g, 13.3 mmol) in EtOH (30 mL). The mixture was heated to 80 °C and stirred for 16 h. Then it was diluted with water (15 mL) and extracted with EA (15 mL × 3). The combined organic phases were dried over Na2SO4, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 2 / 1) to obtain 2-(2-bromo-5-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (2.54 g, yield 46%). LCMS (m / z): [M+H] + C 12 H 10 Calculated for C10H10BrFN3O 309.9; found 310.0.

[0377] Step 5: 2-(5-Fluoro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one: To a solution of 2-(2-bromo-5-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (500 mg, 1.59 mmol) in 1,4-dioxane (10 mL) was added Pd2(dba)3 (85 mg, 0.29 mmol), t-BuXPhos (127 mg, 0.29 mmol), KOH (2.5 mL, 2 mol / L in water) under a nitrogen atmosphere at 100 °C, and microwaved for 1 hour. The mixture was concentrated in vacuo, and the residue was purified by flash column chromatography (petroleum ether:ethyl acetate = 1 / 1) to give 2-(5-fluoro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (227 mg, yield 39%). LCMS (m / z): [M+H] + C 12 H 11 FN3O2 Calculated 248.1; Found 248.1.

[0378] Step 6: To a solution of 2-(5-fluoro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (100 mg, 0.4 mmol) in DMF (2 mL) was added NaHCO3 (102 mg, 1.21 mmol), 2,4-dichloro-5-ethoxymethyl-pyrimidine (126 mg, 0.6 mmol) under a nitrogen atmosphere at 25 °C, and the mixture was stirred at 25 °C for 3 hours. The reaction mixture was filtered, and the filtrate was diluted with water and extracted with EA (4 mL × 3). The combined organic layers were washed with brine (3 mL) and concentrated under reduced pressure. The residue was purified by preparative HPLC to give Product Example 224 (6.26 mg, yield 6.2%). LCMS (m / z): [M+H] + C 19 H 18 ClFN5O3 Calculated 418.1; Found 418.1. 1 1H NMR (400 MHz, DMSO-d6): δ 12.00 (s, 1H), 8.71 (s, 1H), 8.39 (d, J = 2.9 Hz, 1H), 7.66 (d, J = 5.5 Hz, 1H), 7.20 (s, 1H), 7.03 (s, 1H), 4.60 (s, 2H), 3.60 (q, J = 7.0 Hz, 2H), 3.43–3.39 (m, 2H), 2.86 (t, J = 6.8 Hz, 2H), 1.19 (t, J = 7.0 Hz, 3H).

[0379] Test method: Test method B was used.

[0380] Synthesis of Example 225

[0381]

[0382] Step 1: Sodium methoxide (8.55 g, 158.3 mmol, 3.5 equivalents) was added to a solution of ethyl 2,4-dichloropyrimidine-5-carboxylate (10.0 g, 45.2 mmol, 1 equivalent) in MeOH (100 mL), and the reaction was stirred overnight at room temperature. The reaction solvent was removed under reduced pressure, the residue was diluted with EA (100 mL), the mixture was filtered through a Celite pad, and the filtrate was concentrated in vacuo to give methyl 2,4-dimethoxypyrimidine-5-carboxylate as a white solid (6.8 g, 34.3 mmol, 75%). LCMS-ESI (m / z): [M+H] + C8H 10 N2O4 Calculated 199.06; Found 199.03.

[0383] Step 2: DIBALH (68.6 mL, 68.6 mmol, 2 equivalents, 1 M in hexanes) was added to a solution of methyl 2,4-dimethoxypyrimidine-5-carboxylate (6.8 g, 34.3 mmol) in anhydrous DCM (200 mL) at 0 °C under nitrogen, and the reaction was stirred at 0 °C for 1 hour. Water (2.7 mL) was slowly added to the reaction mixture at 0 °C, followed by 15% NaOH solution (2.7 mL) and water (6.8 mL), the mixture was stirred at room temperature for 15 minutes, MgSO4 was added, and the reaction was stirred for another 15 minutes. The residue was filtered and the filtrate was concentrated in vacuo. The mixture was purified by flash column chromatography (MeOH / DCM = 0 - 9%) to give (2,4-dimethoxypyrimidin-5-yl)methanol (3.8 g, 22.3 mmol, 65%). LCMS-ESI (m / z): [M+H] + C7H 10 N2O3 Calculated 171.07; Found 171.09.

[0384] Step 3: To a solution of (2,4-dimethoxypyrimidin-5-yl)methanol (3.8 g, 22.3 mmol) in DCM (120 mL) was added Dess-Martin periodinane (14.2 g, 33.5 mmol, 1.5 equiv.), and the reaction was stirred at room temperature overnight. The reaction was diluted with saturated Na2S2O3 (20 mL) and saturated NaHCO3 (20 mL), the aqueous layer was extracted with DCM (100 mL * 2), the combined organic layers were washed with brine (100 mL), dried over Na2SO4, and purified by flash column (EA / PE = 0 - 15%), to give 2,4-dimethoxypyrimidine-5-carbaldehyde (2.5 g, 15.2 mmol, 45%). LCMS-ESI (m / z): [M+H] + Calculated for C7H8N2O3 169.05; found 169.12.

[0385] Step 4: To a solution of hydroxylamine hydrochloride (1.24 g, 17.8 mmol, 1.2 equiv.) in water (20 mL) was added NaHCO3 (1.5 g, 17.8 mmol, 1.2 equiv.), and the mixture was added dropwise to a solution of 2,4-dimethoxypyrimidine-5-carbaldehyde (2.5 g, 14.8 mmol) in EtOH (40 mL). The reaction was stirred at room temperature for 2 h. The reaction was concentrated, the residue was extracted with DCM (100 mL * 2), the combined organic layers were washed with brine (100 mL) and dried over Na2SO4, to give (E)-2,4-dimethoxypyrimidine-5-carbaldehyde oxime (2.65 g, 14.4 mmol, 98%) without further purification. LCMS-ESI (m / z): [M+H] + Calculated for C7H9N3O3 184.06; found 184.04.

[0386] Step 5: To a solution of (E)-2,4-dimethoxypyrimidine-5-carbaldehyde oxime (2.65 g, 14.4 mmol) in water (50 mL) was added ethynyl(trimethyl)silane (2.13 g, 21.6 mmol, 1.5 equiv.), then KCl (1.08 g, 14.4 mmol, 1 equiv.) was added portionwise, followed by potassium peroxymonosulfate (13.3 g; 21.6 mmol, 1.5 equiv.). The mixture was stirred at room temperature for 12 h. The mixture was extracted with EA (100 mL * 2), the combined organic layers were washed with brine (100 mL), dried over Na2SO4, concentrated, and purified by flash column (EA / PE = 0 - 40%), to give 3-(2,4-dimethoxypyrimidin-5-yl)-5-(trimethylsilyl)isoxazole (2.68 g, 9.6 mmol, 66%). LCMS-ESI (m / z): [M+H] + C 12 H17 Calculated for N3O3Si: 280.10; Found: 279.70.

[0387] Step 6: To a solution of 3-(2,4-dimethoxypyrimidin-5-yl)-5-(trimethylsilyl)isoxazole (2.68 g, 9.6 mmol) in MeOH (50 mL) was added K2CO3 (2.0 g, 14.4 mmol, 1.5 equiv.). The mixture was stirred at room temperature for 0.5 h. The reaction mixture was concentrated and extracted with EA (50 mL × 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, concentrated, and purified by flash column chromatography (EA / PE = 0 - 25%) to give 3-(2,4-dimethoxypyrimidin-5-yl)isoxazole (1.1 g, 5.3 mmol, 55%). LCMS-ESI (m / z): [M+H] + Calculated for C9H9N3O3: 208.06; Found: 207.97.

[0388] Step 7: To a solution of 3-(2,4-dimethoxypyrimidin-5-yl)isoxazole (0.66 g, 3.2 mmol) in AcOH (10 mL) was added NaI (1.43 g, 9.6 mmol, 3 equiv.). The mixture was stirred at 110 °C for 2 h. The reaction mixture was concentrated and diluted with water (20 mL). The residue was washed with saturated Na2S2O3 (20 mL), and the white precipitate was filtered and washed with water to give 3-(2,4-dimethoxypyrimidin-5-yl)isoxazole (0.37 g, 2.0 mmol, 64%). LCMS-ESI (m / z): [M+H] + Calculated for C7H5N3O3: 180.03; Found: 180.06.

[0389] Step 8: To a solution of 3-(2,4-dimethoxypyrimidin-5-yl)isoxazole (0.37 g, 2.0 mmol) in POCl3 (5 mL) was added DIPEA (18 mL, 10 mmol, 5 equiv.). The mixture was stirred at 110 °C for 4 h. The reaction mixture was concentrated and purified by flash column chromatography (EA / PE = 0 - 15%) to give 3-(2,4-dichloropyrimidin-5-yl)isoxazole as yellow crystals (0.38 g, 1.75 mmol, 87%). LCMS-ESI (m / z): [M+H] + Calculated for C7H3Cl2N3O: 215.97; Found: 216.00.

[0390] Step 9: To a solution of 3-(2,4-dichloropyrimidin-5-yl)isoxazole (44 mg, 0.19 mmol) in DMAc (1 mL) was added Int-0012 (50 mg, 0.22 mmol, 1.2 equiv) and potassium tert-butoxide (32 mg, 0.29 mmol, 1.5 equiv), and the mixture was stirred at 55 °C for 2 h. The mixture was concentrated and purified by flash column chromatography to afford Example 225 as a yellow solid (5.4 mg, 0.012 mmol, 7%). LCMS-ESI (m / z): [M+H] + C 19 H 13 ClN6O3 Calculated 409.07; Found 408.96. 1 1H NMR (600 MHz, DMSO-d6): δ 12.02 (s, 1H), 9.20 (s, 1H), 9.18 (d, J = 2.0 Hz, 1H), 8.30 (d, J = 5.2 Hz, 1H), 7.67 - 7.64 (m, 1H), 7.62 - 7.59 (m, 1H), 7.17 (d, J = 1.6 Hz, 1H), 7.14 - 7.10 (m, 2H), 3.45 - 3.38 (m, 2H), 2.85 (t, J = 6.8 Hz, 2H).

[0391] Test method: Test method A used; retention time = 2.09 min

[0392] Synthesis of Example 226

[0393]

[0394] Step 1: To a solution of 2,4-dichloro-5-(iodomethyl)pyrimidine (1.00 g, 3.47 mmol) and DIEA (888 mg, 6.94 mmol) in dichloromethane (10 mL) at -20 °C was added dropwise piperidine (295 mg, 3.47 mmol) in DCM (2 mL). The reaction mixture was stirred at -20 °C for 3 h. The reaction mixture was extracted with DCM (30 mL), washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a crude product which was purified by flash column chromatography on silica gel (EA:DCM = 1 / 1) to afford 2,4-dichloro-5-(piperidin-1-ylmethyl)pyrimidine as a yellow oil (0.3 g, yield 35%). LCMS (m / z): [M+H] + C10H14Cl2N3 Calculated 246.05; Found 246.1.

[0395] Step 2: At room temperature, add NaHCO3 (144 mg, 1.72 mmol) to a solution of 2,4-dichloro-5-(piperidin-1-ylmethyl)pyrimidine (107 mg, 0.43 mmol) and Int-0012 (100 mg, 0.43 mmol) in DMF (5 mL). Stir the reaction mixture at 40 °C for 16 hours. Extract the reaction mixture with EA (20 mL * 2), wash it with brine (10 mL * 3), dry it over Na2SO4, filter and concentrate it under reduced pressure to obtain the crude product. Purify the crude product by preparative HPLC to obtain the product Example 226 (17 mg, yield 17%) as a yellow solid. LCMS (m / z): [M+H] + C 22 H 24 ClN6O2 Calculated 439.16; Found 438.9. 1 H NMR (400 MHz, DMSO) δ 12.07 (s, 1H), 9.71 (s, 1H), 8.86 (s, 1H), 8.32 (d, J = 5.4 Hz, 1H), 7.67–7.63 (m, 1H), 7.58 (s, 1H), 7.13 (d, J = 2.1 Hz, 2H), 4.45 (s, 2H), 3.53 (d, J = 11.2 Hz, 2H), 3.41 (t, J = 6.4 Hz, 2H), 3.14–3.02 (m, 2H), 2.86 (t, J = 6.8 Hz, 2H), 1.85 (d, J = 13.3 Hz, 2H), 1.68 (t, J = 10.8 Hz, 3H), 1.40 (t, J = 11.5 Hz, 1H).

[0396] Test method: Test method B used. Retention time = 3.479 min, 3.601 min

[0397] Synthesis of Example 227

[0398]

[0399] Step 1: A mixture of Int-008 (2.0 g, 4.32 mmol, 1.0 equiv), 2-bromo-1-(2-chloropyrimidin-4-yl)ethan-1-one (1.02 g, 4.32 mmol, 1.0 equiv), and NH4OAc (999 mg, 12.96 mmol, 3 equiv) in EtOH (50 mL) was stirred at 60 °C overnight. The mixture was concentrated and purified by silica gel column chromatography, eluting with DCM and MeOH (20:1) to give tert-butyl 2'-(2-chloropyrimidin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (1.94 g, 75%). LCMS-ESI (m / z): [M+H] + C 30 H 36 Calculated for ClN5O6 598.10; found 598.10.

[0400] Step 2: TFA (5 mL) was added to a solution of tert-butyl 2'-(2-chloropyrimidin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (1.94 g, 3.24 mmol, 1.0 equiv) in DCM (20 mL), and the reaction was stirred at 50 °C for 2 h. The mixture was concentrated to give the crude 2'-(2-chloropyrimidin-4-yl)-5',6'-dihydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-4'(1'H)-one. LCMS-ESI (m / z): [M+H] + C 15 H 16 Calculated for ClN5O 317.78; found 317.78.

[0401] Step 3: TEA (5 mL), DMAP (33 mg, 0.32 mmol, 0.1 equiv), and (Boc)2O (1.41 g, 6.48 mmol, 2 equiv) were added to a solution of the crude 2'-(2-chloropyrimidin-4-yl)-5',6'-dihydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-4'(1'H)-one in MeCN (50 mL), and the reaction was stirred at room temperature overnight. The mixture was concentrated and purified by silica gel column chromatography, eluting with DCM and MeOH (20:1) to give tert-butyl 2'-(2-chloropyrimidin-4-yl)-4'-oxo-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (1.02 g, 75% over 2 steps). LCMS-ESI (m / z): [M+H]+ C 20 H 24 Calculated for C + H 20 ClN5O3: 417.89; Found: 417.89.

[0402] Step 4: A mixture of tert-butyl 2'-(2-chloropyrimidin-4-yl)-4'-oxo-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (1.02 g, 2.44 mmol, 1.0 equiv), N-hydroxyacetamide (549 mg, 7.31 mmol, 3.0 equiv) and K2CO3 (1.68 g, 12.18 mmol, 5.0 equiv) in DMSO (8.1 mL) was stirred overnight at 80 °C under nitrogen. The mixture was concentrated and purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (1:1) to afford tert-butyl 2'-(2-hydroxypyrimidin-4-yl)-4'-oxo-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (405 mg, 42%). LCMS-ESI (m / z): [M+H] + + C 20 H 25 Calculated for C + H 20 N5O4: 399.45; Found: 399.45.

[0403] Step 5: To a solution of tert-butyl 2'-(2-hydroxypyrimidin-4-yl)-4'-oxo-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (200 mg, 0.50 mmol, 1.0 equiv) in DMF (1 mL) was added 2,4-dichloro-5-(ethoxymethyl)pyrimidine (109 mg, 0.53 mmol, 1.05 equiv) and NaHCO3 (85 mg, 1.00 mmol, 2.0 equiv). The reaction mixture was stirred at 100 °C for 2 h. The reaction mixture was concentrated and purified by preparative HPLC to afford Example 227 (24 mg, 8%). LCMS-ESI (m / z): [M+H] + + C 27 H 32 Calculated for C + H 27 ClN7O5: 570.05; Found: 570.05. 11H NMR (DMSO-d6): δ 12.03 (s, 1H), 8.80 (m, 1H), 8.64 (d, J = 5.4 Hz, 1H), 7.86 (d, J = 5.4 Hz, 1H), 7.38 (s, 1H), 7.33 (s, 1H), 4.58 (s, 2H), 4.08 - 3.92 (m, 2H), 3.58 - 3.54 (m, 2H), 3.32 - 3.20 (m, 2H), 2.42 - 2.34 (m, 2H), 1.73 - 1.71 (m, 1H), 1.59 - 1.57 (m, 1H), 1.45 - 1.41 (m, 2H), 1.41 (s, 9H), 1.15 (t, J = 7.2 Hz, 3H).

[0404] Test method: Test method A used. Retention time = 2.58 minutes

[0405] Synthesis of Example 228

[0406]

[0407] Step 1: To a solution of Int-009 (50 mg, 0.50 mmol, 1.0 equiv) in DMF (2 mL) was added 2,4-dichloro-5-(ethoxymethyl)pyrimidine (45 mg, 0.25 mmol, 2.0 equiv) and NaHCO3 (20 mg, 0.25 mmol, 2.0 equiv). The reaction was stirred at 100 °C for 2 h. The reaction mixture was concentrated and purified by preparative HPLC to give Example 228 (16 mg, 22%). LCMS-ESI (m / z): [M + H] + C 27 H 32 ClN7O5 Calculated 569.06; Found 569.06. 11H NMR (600 MHz, DMSO-d6): δ 11.60 (d, J = 15.3 Hz, 2H), 8.71 (d, J = 5.3 Hz, 2H), 8.31 (d, J = 5.4 Hz, 1H), 8.27 (d, J = 5.4 Hz, 1H), 7.73 (dd, J = 5.4 Hz, 1.4 Hz, 1H), 7.69 (dd, J = 5.4 Hz, 1.4 Hz, 1H), 7.63 (s, 1H), 7.61 (s, 1H), 7.25 (s, 2H), 7.14 (dd, J = 10.7 Hz, 2.2 Hz, 2H), 4.60 (s, 2H), 4.53 (s, 2H), 4.06 (d, J = 28.8 Hz, 4H), 3.62 - 3.55 (m, 4H), 3.30 (d, J = 3.5 Hz, 2H), 3.20 (d, J = 12.4 Hz, 2H), 2.99 (s, 2H), 2.68 (s, 2H), 2.22 - 2.20 (m, 2H), 1.78 (d, J = 13.0 Hz, 2H), 1.62 (d, J = 13.3 Hz, 2H), 1.54 - 1.46 (m, 2H), 1.47 - 1.41 (m, 17H), 1.21 - 1.17 (m, 6H).

[0408] Test method: Test method A used. Retention time = 2.56 minutes

[0409] Synthesis of Example 229

[0410]

[0411] Step 1: To a solution of Int-009 (84 mg, 0.21 mmol, 1.0 equiv) in DMF (1 mL) was added 2,4-dichloropyrimidine-5-carboxamide (81 mg, 0.42 mmol, 2.0 equiv) and NaHCO3 (35 mg, 0.42 mmol, 2.0 equiv), and the reaction was stirred at 100 °C for 2 h. The reaction mixture was concentrated and purified by preparative HPLC to give Example 229 (10 mg, 8%). LCMS-ESI (m / z): [M+H] + C 26 H 28 ClN7O5 Calculated 554.18; Found 553.98. 11H NMR (600 MHz, DMSO-d6): δ 11.59 (s, 1H), 8.94 (s, 1H), 8.32 (d, J = 5.4 Hz, 1H), 7.96 (s, 1H), 7.93 (s, 1H), 7.75 - 7.72 (m, 1H), 7.66 - 7.63 (m, 1H), 7.20 (s, 1H), 7.12 (d, J = 2.4 Hz, 1H), 4.14 - 3.98 (m, 2H), 3.34 - 3.29 (m, 1H), 3.23 - 3.18 (m, 1H), 2.98 (brs, 1H), 2.70 (brs, 1H), 2.26 - 2.16 (m, 1H), 1.81 - 1.76 (m, 1H), 1.66 - 1.60 (m, 1H), 1.55 - 1.45 (m, 1H), 1.42 (s, 9H).

[0412] Test method: Test method A used. Retention time = 2.24 minutes

[0413] Synthesis of Example 230

[0414]

[0415] Step 1: To a solution of Int-0010 (150 mg, 0.23 mmol) in dioxane (1.5 mL) was added 6-chloropicolinamide (55 mg, 0.35 mmol), Xantphos (27 mg, 0.047 mmol), Pd2(dba)3 (21 mg, 0.023 mmol) and Cs2CO3 (228 mg, 0.70 mmol). The reaction mixture was stirred under microwave at 100 °C for 1 hour. The reaction was quenched with water (5 mL), the solid was collected by filtration, washed with water (3 mL), and then freeze-dried to give tert-butyl 2'-(2-(6-chloropicolinamido)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (110 mg, yield: 59%). LCMS (m / z): [M+H] + Calculated for C37H42ClN6O7 717.3; found 717.3.

[0416] Step 2: A solution of tert-butyl 2'-(2-(6-chloropyridinecarboxamido)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (110 mg, 0.15 mmol) and anisole (83 mg, 0.77 mmol) in TFA (1 mL) was stirred at 25 °C for 30 minutes. After evaporation, the crude product was purified by preparative HPLC (AQ-C18, ACN---H2O (0.1% TFA)), flow rate: 45 ml / min; wavelength: 214 nm. The sample in DMSO was injected, with a linear gradient from 30% to 80% ACN over 12.5 minutes, to give Compound Example 230 (14.7 mg, 21%). LCMS (m / z): [M+H] + Calculated for C22H22ClN6O2 437.1; Found 437.1. 1 H NMR (400 MHz, DMSO) δ 11.86 (s, 1H), 10.25 (s, 1H), 9.08 (d, J = 10.5 Hz, 1H), 8.50 (m, 2H), 8.39 (d, J = 5.4 Hz, 1H), 8.28–8.13 (m, 2H), 7.97–7.82 (m, 1H), 7.57 (d, J = 5.3 Hz, 1H), 7.42 (s, 1H), 6.99 (t, J = 4.0 Hz, 1H), 3.61 (d, J = 10.6 Hz, 1H), 3.51–3.21 (m, 4H), 2.78 (m, 1H), 2.15 (t, J = 11.7 Hz, 1H), 1.87 (d, J = 12.1 Hz, 2H), 1.72 (m, 1H).

[0417] Synthesis of Example 231

[0418]

[0419] Step 1: At room temperature, K2CO3 (215 mg, 1.56 mmol) was added to a solution of Int-009 (300 mg, 519 μmol, 1.0 equiv) and 2,4-dichloro-5-(ethoxymethyl)pyrimidine (128 mg, 623 μmol, 1.2 equiv) in acetonitrile (10 mL). The mixture was stirred at 60 °C for 2 hours and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:dichloromethane = 5 / 1) to give tert-butyl 2'-(2-((2-chloro-5-(ethoxymethyl)pyrimidin-4-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (40 mg, yield 10%). LCMS (m / z): [M+H] + C 38 H 45 Calculated for ClN6O8: 749.3; Found: 749.3.

[0420] Step 2: At 25 °C, TFA (2 mL) was added to a solution of tert-butyl 2'-(2-((2-chloro-5-(ethoxymethyl)pyrimidin-4-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (40 mg, 53.4 μmol) in anisole (1 mL), and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was quenched with H2O. The aqueous phase was lyophilized to give the crude product in the form of the FA salt. The crude product was purified by preparative HPLC to give Example 231 (4.06 mg, yield 16%). LCMS (m / z): [M+H] + C 23 H 25 Calculated for ClN6O3: 469.2; Found: 469.3. 1H NMR (400 MHz, CD3OD): δ 8.68 - 8.64 (m, 1H), 8.36 (d, J = 4.0 Hz, 1H), 7.72 - 7.67 (m, 1H), 7.60 - 7.55 (m, 1H), 7.19 (s, 1H), 4.70 (s, 2H), 3.75 - 3.68 (m, 2H), 3.62 - 3.46 (m, 3H), 3.31–3.21 (m, 2H), 3.06 - 2.98 (m, 1H), 3.35 - 3.25 (m, 1H), 2.14 - 1.93 (m, 3H), 1.30 (t, J = 8.0 Hz, 3H).

[0421] Test method: Test method B was used. Retention time = 3.898 minutes

[0422] Synthesis of Example 232 / 233

[0423]

[0424] Step 1: At 80 °C under nitrogen, add 2,4-dichloro-5-(((4-methoxybenzyl)oxy)methyl)pyrimidine (434 mg, 1.45 mmol) and K2CO3 (501 mg, 3.62 mmol) to a solution of Int-007 (700 mg, 1.21 mmol) in acetonitrile (7 mL). Stir the resulting mixture at 80 °C for 4 hours, then filter. Concentrate the filtrate in vacuo and purify the residue by preparative TLC (dichloromethane:methanol = 20 / 1) to obtain tert-butyl 2'-(2-((2-chloro-5-(((4-methoxybenzyl))oxy)methyl)pyrimidin-4-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (45 mg, yield 4%), LCMS (m / z): [M+H] + C 44 H 49 ClN6O9 Calcd 841.3; Found 841.2, and obtain tert-butyl 2'-(2-((4-chloro-5-((4-methoxybenzyl)oxy)pyrimidin-2-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (22 mg, yield 2%), LCMS (m / z): [M+H] + C 44 H 49 ClN6O9 Calcd 841.3; Found 841.2.

[0425] Step 2a: At room temperature, anisole (30 mg, 0.28 mmol) was added to a solution of tert-butyl 2'-(2-((2-chloro-5-(((4-methoxybenzyl)oxy)methyl)pyrimidin-4-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (45 mg, 0.054 mmol) in TFA (1 mL), and the mixture was stirred for 1 hour. The mixture was concentrated in vacuo, and the residue was purified by preparative HPLC (column: Gemini 5u C18 150x 21.2 mm, ACN-H2O (0.1% TFA), flow rate: 20 ml / min; wavelength: 214 nm / 254 nm, 8.31-minute gradient maintained at 20% CH3CN), to give Example 232 (12.11 mg, yield 24%). LCMS (m / z): [M+H] + C 21 H 21 Calculated for ClN6O3 441.1; found 441.2. 1 H NMR (400 MHz, MeOD) δ 8.62 (s, 1H), 8.30 (d, J = 5.5 Hz, 1H), 7.65 (dd, J = 5.5, 1.6 Hz, 1H), 7.52 (d, J = 1.2 Hz, 1H), 7.13 (s, 1H), 4.76 (s, 2H), 3.70 (d, J = 13.3 Hz, 1H), 3.60–3.39 (m, 3H), 3.25 (d, J = 13.2 Hz, 1H), 2.97 (m, 1H), 2.26 (m, 1H), 2.09–1.81 (m, 3H).

[0426] Step 2b: At room temperature, anisole (15 mg, 0.14 mmol) was added to a solution of tert-butyl 2'-(2-((4-chloro-5-((4-methoxybenzyl)oxy)pyrimidin-2-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (22 mg, 0.027 mmol) in TFA (1 mL), and the mixture was stirred for 1 hour. The mixture was concentrated in vacuo, and the residue was purified by preparative HPLC (column: Gemini5u C18 150x 21.2 mm, ACN-H2O (0.1% TFA), flow rate: 20 ml / min; wavelength: 214 nm / 254 nm, 8.11-minute gradient maintained at 20% CH3CN), to give Example 233 (1.91 mg, yield 17%). LCMS (m / z): [M+H] +C 21 H 21 Calculated for C₂₀H₂₀ClN₆O₃: 441.1; Found: 441.2. 1 ¹H NMR (400 MHz, MeOD) δ 8.63 (s, 1H), 8.26 (d, J = 5.4 Hz, 1H), 7.60 (d, J = 5.4 Hz, 1H), 7.50 (s, 1H), 7.13 (s, 1H), 4.67 (s, 2H), 3.69 (d, J = 13.2 Hz, 1H), 3.61–3.38 (m, 3H), 3.22 (m, 1H), 2.97 (m, 1H), 2.31–2.17 (m, 1H), 2.12–1.78 (m, 3H).

[0427] Test method: Test method B was used. Example 232, retention time = 3.31 minutes; Example 233, retention time = 3.32 minutes

[0428] Synthesis of Example 234

[0429]

[0430] Step 1: To a solution of Int-009 (500 mg, 0.86 mmol) in acetonitrile (5 mL) at 80 °C under nitrogen was added 4,6-dichloropyrimidine (141 mg, 0.95 mmol) and K₂CO₃ (358 mg, 2.59 mmol). The resulting mixture was stirred at 80 °C for 16 h and then filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (dichloromethane:methanol = 20 / 1) to afford tert-butyl 2'-(2-((6-chloropyrimidin-4-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (300 mg, yield 45%), LCMS (m / z): [M+H] + C 35 H 39 Calculated for C₃₀H₃₀ClN₆O₇: 691.2; Found: 691.2.

[0431] Step 2: To a solution of tert-butyl 2'-(2-((6-chloropyrimidin-4-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (100 mg, 0.14 mmol) in TFA (1 mL) was added anisole (78 mg, 0.72 mmol), and the mixture was stirred for 1 h. The mixture was concentrated in vacuo, and the residue was purified by preparative HPLC (column: Gemini 5u C18 150x21.2 mm, ACN-H2O (0.1% TFA), flow rate: 20 ml / min; wavelength: 214 nm / 254 nm, 8.0 min gradient maintained at 30% CH3CN) to give Example 234 (19.04 mg, 32% yield). LCMS (m / z): [M+H] + C 20 H 19 Calculated for C21H22ClN6O2 411.1; found 411.0. 1 1H NMR (400 MHz, MeOD) δ 8.64 (s, 1H), 8.33 (d, J = 5.4 Hz, 1H), 7.67 (dd, J = 5.4, 1.5 Hz, 1H), 7.55 (s, 1H), 7.38 (s, 1H), 7.17 (s, 1H), 3.74 (d, J = 13.3 Hz, 1H), 3.67–3.42 (m, 3H), 3.28 (d, J = 13.3 Hz, 1H), 3.01 (m, 1H), 2.29 (m, 1H), 2.13–1.85 (m, 3H).

[0432] Test method: Test method B was used. Retention time = 3.66 min

[0433] Synthesis of Example 235

[0434]

[0435] Step 1: To a solution of 2,4-dichloropyrimidine-5-carbonitrile (77 mg, 0.44 mmol, 1.2 equiv) and Int-0012 (85 mg, 0.37 mmol, 1.0 equiv) in DMAc (10 mL) was added NaHCO3 (37 mg, 0.44 mmol, 1.2 equiv). The reaction was stirred at 50 °C for 2 h. The reaction mixture was purified by preparative HPLC to give Example 235 (3.9 mg, 18 μmol, 5%). LCMS-ESI (m / z): [M+H] + C 17 H 11 Calculated for C18H13ClN6O2 367.06; found 366.99.

[0436] 1 1H NMR (DMSO-d6): δ 12.06 (s, 1H), 9.31 (s, 1H), 8.34 (d, J = 5.2 Hz, 1H), 7.71 - 7.69 (dd, J = 1.6 Hz, 5.2 Hz, 1H), 7.63 (d, J = 0.8 Hz, 1H), 7.12 (m, 2H), 3.43 - 3.39 (m, 2H), 2.87 - 2.84 (m, 2H).

[0437] Test method: Test method A used. Retention time = 2.043 minutes

[0438] Synthesis of Example 236

[0439]

[0440] Step 1: To a solution of Int-0013 (600 mg, 1.46 mmol, 1.0 equiv) and pyridin-4-ylboronic acid (269 mg, 2.19 mmol, 1.5 equiv) in 1,4-dioxane (30 mL) and water (6 mL) was added Pd(dppf)Cl2-DCM (119 mg, 146 μmol, 0.1 equiv) and NaHCO3 (368 mg, 4.38 mmol, 3.0 equiv). The reaction was stirred at 55 °C overnight. The mixture was filtered, the solution was concentrated to give the crude product. The crude product was purified by silica gel column chromatography to give 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-(pyridin-4-yl)pyrimidine (50 mg, 0.14 mmol, 10%). LCMS-ESI (m / z): [M+H] + C 14 H8BrClN4O Calculated 362.96; Found 362.89.

[0441] Step 2: To a solution of 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-(pyridin-4-yl)pyrimidine (180 mg, 0.50 mmol, 1.0 equiv) and 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (262 mg, 1 mmol, 2.0 equiv) in 1,4-dioxane (10 mL) and water (2 mL) was added Pd(dppf)Cl2-DCM (41 mg, 50 μmol, 0.1 equiv) and K3PO4 (318 mg, 1.5 mmol, 3.0 equiv). The reaction was stirred in a microwave at 60 °C for 1 h. The mixture was filtered and the solution was concentrated to give the crude product. The residue was purified by preparative HPLC to give Example 236 (3.8 mg, 9 μmol, 2%). LCMS-ESI (m / z): [M+H] + C 21 H 15 ClN6O2 Calculated 419.09; Found 418.89. 1 1H NMR (DMSO-d6): δ 12.01 (s, 1H), 8.98 (s, 1H), 8.75 - 8.73 (dd, J = 1.6 Hz, 4.4 Hz, 2H), 8.29 (d, J = 5.6 Hz, 1H), 7.78 - 7.77 (dd, J = 1.6 Hz, 4.4 Hz, 2H), 7.64 - 7.62 (m, 1H), 7.60 (s, 1H), 7.12 (d, J = 2.4 Hz, 2H), 3.43 - 3.40 (m, 2H), 2.86 - 2.83 (m, 2H).

[0442] Test method: Test method A used. Retention time = 1.817 min

[0443] Synthesis of Example 237

[0444]

[0445] Step 1: To a solution of 2-bromo-1-(2-chloropyridin-4-yl)ethanone (10 g, 42.6 mmol) in EtOH (50 mL) was added NH4AC (19.7 g, 255.6 mmol), HOAc (10.23 g, 170.4 mmol) and tert-butyl 2,4-dioxopiperidine-1-carboxylate (18.17 g, 85.2 mmol). The resulting mixture was stirred at 80 °C for 16 h under a nitrogen atmosphere. The mixture was diluted with water (200 mL) and filtered. The filter cake was tert-butyl 2-(2-chloropyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (12.5 g, yield 63%). It can be used for the next step without further purification. LCMS (m / z): [M+H] + Calculated 348.1; found 348.1.

[0446] Step 2: At 25 °C under a nitrogen atmosphere, to a solution of tert-butyl 2-(2-chloropyridin-4-yl)-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (4 g, 11.5 mmol) in DMF (25 mL) was added methyl iodide (8.14 g, 57.3 mmol) and potassium carbonate (1.93 g, 34.4 mmol) over 1 h. The mixture was diluted with water (60 mL) and extracted with EtOAc (30 mL×3). The organic layer was washed with brine (40 mL) and concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 10 / 1) to give tert-butyl 2-(2-chloropyridin-4-yl)-1-methyl-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (2.65 g, yield 41%). LCMS (m / z): [M+H]+ Calculated 362.1; found 362.1.

[0447] Step 3: At 130 °C, to a solution of tert-butyl 2-(2-chloropyridin-4-yl)-1-methyl-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (2.65 g, 7.3 mmol) in AcOH (18 mL) was added H2O (6 mL) over 120 h. The reaction mixture was concentrated in vacuo and the residue was purified by C 18 reverse-phase column chromatography (H2O:ACN = 6 / 1) to give 2-(2-hydroxypyridin-4-yl)-1-methyl-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one as a yellow solid (1.13 g, yield 29%). LCMS (m / z): [M+H] + Calculated 244.1; found 244.1.

[0448] Step 4: At 25 °C under a nitrogen atmosphere, 2,4-dichloro-5-iodopyrimidine (539 mg, 1.9 mmol) and NaHCO3 (329 mg, 3.9 mmol) were added to a solution of 2-(2-hydroxypyridin-4-yl)-1-methyl-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (300 mg, 1.3 mmol) in DMF (5 mL). Then the mixture was heated to 90 °C and stirred for 16 h. The mixture was diluted with water (15 mL) and extracted with EA (15 mL × 3). The organic layer was washed with brine (10 mL) and concentrated under reduced pressure. The residue was purified by C18 reverse-phase column chromatography (H2O:ACN = 4 / 1) to give 2-(2-((2-chloro-5-iodopyrimidin-4-yl)oxy)pyridin-4-yl)-1-methyl-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (270 mg, yield 31%). LCMS (m / z): [M+H]+ calculated 481.9; found 482.0.

[0449] Step 5: A solution of 2-(2-((2-chloro-5-iodopyrimidin-4-yl)oxy)pyridin-4-yl)-1-methyl-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (130 mg, 0.27 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (63 mg, 0.32 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (20 mg, 0.027 mmol), and tripotassium phosphate (172 mg, 0.81 mmol) in 1,4-dioxane (1.5 mL) and H2O (0.15 mL) was stirred at 65 °C under a nitrogen atmosphere for 2 h. The reaction mixture was filtered and the filtrate was purified by preparative HPLC to give Example 237 (0.83 mg, yield 0.72%). LCMS (m / z): [M+H]+ calculated 423.09; found 423.1. 1 H NMR (400 MHz, DMSO-d6): δ 9.58 (s, 1H), 9.33 (s, 1H), 9.15 (s, 1H), 8.35 (d, J = 5.8 Hz, 1H), 7.53–7.51 (m, 2H), 7.08 (s, 1H), 6.75 (s, 1H), 3.66 (s, 3H), 2.84 (t, J = 6.9 Hz, 2H), 2.29 (t, J = 3.7 Hz, 2H).

[0450] Test method: Test method D was used.

[0451] Synthesis of Example 238

[0452]

[0453] Step 1: A suspension of Int-009 (300 mg, 0.52 mmol), 2,4-dichloro-5-iodopyrimidine (170 mg, 0.62 mmol) and NaHCO3 (130 mg, 1.5524 mmol) in DMF (5 mL) was stirred under nitrogen at 90 °C for 2 h. The residue was diluted with EtOAc (20 mL × 2) and washed with brine (20 mL). The organic phase was collected, dried over NaSO44 and concentrated. The residue was purified by flash column chromatography (petroleum ether:ethyl acetate = 2 / 1) to give tert-butyl 2'-(2-((2-chloro-5-iodopyrimidin-4-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (360 mg, yield 85%). LCMS (m / z): [M+H]+ calculated 817.1; found 817.1.

[0454] Step 2: A suspension of tert-butyl 2'-(2-((2-chloro-5-iodopyrimidin-4-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (260 mg, 0.32 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (123 mg, 0.64 mmol), Pd(dppf)Cl2 (37 mg, 0.03 mmol) and K2CO3 (220 mg, 1.59 mmol) in dioxane / H2O (5 mL) was stirred under nitrogen at 60 °C for 2 h. The reaction mixture was concentrated under reduced pressure, and the residue was purified by C18 reverse-phase column chromatography (H2O:ACN = 6 / 1) to give tert-butyl 2'-(2-((2-chloro-5-(isoxazol-4-yl)pyrimidin-4-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (140 mg, yield 52%). LCMS (m / z): [M+H]+ calculated 758.2; found 758.0.

[0455] Step 3: A solution of tert-butyl 2'-(2-((2-chloro-5-(isoxazol-4-yl)pyrimidin-4-yl)oxy)pyridin-4-yl)-4'-oxo-5'-(2,4,6-trimethoxybenzyl)-1',4',5',6'-tetrahydrospiro[piperidine-3,7'-pyrrolo[3,2-c]pyridine]-1-carboxylate (140 mg, 0.18 mmol) in anisole (3 mL) and TFA (3 mL) was stirred at 25 °C for 3 h. The mixture was concentrated and the residue was treated with 1N HCl solution (5 mL) and EtOAc (5 mL). After separation, the aqueous phase was lyophilized. The residue was purified by preparative HPLC (column: Gemini 5u C18 150x 21.2 mm, ACN-H2O (0.1% TFA), flow rate: 20 ml / min; wavelength: 214 nm / 254 nm, 8.37 min gradient maintained at 70% CH3CN) to give Example 238 (HCl salt, 4.59 mg, yield 5%). LCMS (m / z): [M+H]+ calculated 478.1; found 478.1. 1 H NMR (400 MHz, d6-DMSO) δ 9.64 (s, 1H), 9.38 (s, 1H), 9.20 (s, 1H), 8.34 (d, J = 5.3 Hz, 1H), 8.17 (s, 1H), 7.76 (d, J = 5.3 Hz, 1H), 7.71 (s, 1H), 7.23 (s, 1H), 7.13 (s, 1H), 3.70–3.63 (m, 1H), 3.08–2.95 (m, 2H), 2.93–2.84 (m, 1H), 2.69–2.63 (m, H), 2.14.

[0456] Test method: Test method B was used. Retention time = 3.68 min.

[0457] Synthesis of Example 239

[0458]

[0459] Step 1: To a solution of Int-0013 (110 mg, 0.23 mmol) in dioxane (6 mL) and water (0.6 mL) was added K3PO4 (100 mg, 0.47 mmol), 5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (59 mg, 0.28 mmol), and Pd(dppf)Cl2 (17 mg, 0.023 mmol). The mixture was stirred at 50 °C under a nitrogen atmosphere for 4 h. The mixture was filtered and the filtrate was purified by preparative HPLC using a H2O(0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Sample was injected in DMF, linear gradient from 25% to 40% ACN in 12.5 min), to give Example 239 (59.79 mg, 59%). LCMS (m / z): [M+H] + Calculated 423.0; found 423.1. 1 1H NMR (400 MHz, d6-DMSO) δ 12.03 (s, 1H), 8.86 (s, 1H), 8.84 (s, 1H), 8.28 (d, J = 5.4 Hz, 1H), 7.63 (dd, J = 5.4, 1.3 Hz, 1H), 7.58 (s, 1H), 7.11 (d, J = 2.2 Hz, 2H), 3.41 (t, J = 6.8 Hz, 2H), 2.85 (t, J = 6.8 Hz, 2H), 2.58 (s, 3H).

[0460] Test method: Test method B was used. Retention time = 4.154 min.

[0461] Synthesis of Example 240

[0462]

[0463] Step 1: To a solution of Int-0013 (100 mg, 0.21 mmol) in 1,4-dioxane (2 mL) and H2O (0.2 mL) at 50 °C under a nitrogen atmosphere was added 1-methyl-1H-pyrazole-4-boronic acid (32 mg, 0.25 mmol), Pd(dppf)Cl2 (16 mg, 0.02 mmol), and potassium carbonate (89 mg, 0.64 mmol) for 2 h. The reaction mixture was filtered and the filtrate was concentrated. The residue was purified by preparative HPLC to give Example 240 (27.26 mg, yield 27%). LCMS (m / z): [M+H]+ Calculated 422.1; found 421.7. 11H NMR (400 MHz, d6-DMSO): δ 11.95 (s, 1H), 9.06 (s, 1H), 8.33 (s, 1H), 8.25 (d, J = 5.4 Hz, 1H), 8.09 (s, 1H), 7.60 (dd, J = 5.4, 1.5 Hz, 1H), 7.54 (d, J = 1.2 Hz, 1H), 7.09 (d, J = 2.4 Hz, 1H), 7.07 (d, J = 2.3 Hz, 1H), 3.87 (s, 3H), 3.37 (dd, J = 6.8, 4.5 Hz, 2H), 2.81 (t, J = 6.8 Hz, 2H).

[0464] Test method: Test method D was used.

[0465] Synthesis of Example 241

[0466]

[0467] Step 1: At 50 °C under a nitrogen atmosphere, 2-fluorophenylboronic acid (24 mg, 0.17 mmol), Pd(dppf)Cl2 (16 mg, 0.02 mmol), and potassium carbonate (89 mg, 0.64 mmol) were added to a solution of Int-0013 (100 mg, 0.21 mmol) in 1,4-dioxane (2 mL) and H2O (0.2 mL) for 16 hours. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC to obtain Example 241 (15.35 mg, yield 15%). LCMS (m / z): [M+H]+ calculated 436.09; found 436.1. 1 1H NMR (400 MHz, d6-DMSO): δ 11.98 (s, 1H), 8.83 (s, 1H), 8.24 (d, J = 5.4 Hz, 1H), 7.63 (ddd, J = 6.9, 6.2, 1.1 Hz, 3H), 7.49 (s, 1H), 7.43 - 7.35 (m, 2H), 7.11 (s, 1H), 7.10 (s, 1H), 3.40 (dd, J = 6.6, 4.3 Hz, 2H), 2.84 (t, J = 6.9 Hz, 2H).

[0468] Test method: Test method D was used.

[0469] Synthesis of Example 242

[0470]

[0471] Step 1: At room temperature, Pd(PPh3)4 (22 mg, 0.019 mmol) and K2CO3 (123 mg, 0.58 mmol) were added to a solution of Int-0013 (90 mg, 0.19 mmol, 1 equiv) and 2 (29 mg, 0.21 mmol, 1.1 equiv) in dioxane (5 mL) and water (1 mL). The mixture was stirred at 65 °C for 4 h and then filtered. The filtered mixture was purified by preparative HPLC using a H2O(0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample was injected in DMF, with a 50-minute linear gradient from 10% to 90% ACN), to give Example 242 (6.17 mg, 7%). LCMS (m / z): [M+H]+ calculated 433.0; found 433.1. 1 1H NMR (400 MHz, d6-DMSO) δ 12.02 (s, 1H), 8.91 (s, 1H), 8.78 (d, J = 2.2 Hz, 1H), 8.27 (d, J = 5.4 Hz, 1H), 8.04 (dd, J = 8.1, 2.4 Hz, 1H), 7.62 (dd, J = 5.4, 1.4 Hz, 1H), 7.57 (s, 1H), 7.43 (d, J = 8.1 Hz, 1H), 7.14–7.07 (m, 2H), 3.43–3.39 (m, 2H), 2.85 (t, J = 6.8 Hz, 2H), 2.53 (s, 3H).

[0472] Test method: Test method B was used. Retention time = 3.2 minutes.

[0473] Synthesis of Example 243

[0474]

[0475] Step 1: At room temperature in a glove box, purified water (1 mL, 2V), Pd(dppf)Cl2 (12 mg, 0.017 mmol) and K3PO4 (109 mg, 0.51 mmol) were added to a solution of Int-0013 (80 mg, 0.17 mmol, 1 equiv) and 2 (38 mg, 0.34 mmol, 2.0 equiv) in dioxane (5 mL, 20V). The mixture was stirred at 65 °C for 4 h and then filtered. The filtered mixture was purified by preparative HPLC using a H2O(0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample was injected in DMF, with a 50-minute linear gradient from 10% to 90% ACN), to give Example 243 (3.29 mg, 4%). LCMS (m / z): [M+H]+ calculated 407.0; found 408.1.1 1H NMR (400 MHz, d6-DMSO) δ 13.39 (s, 1H), 12.05 (s, 1H), 9.19 (s, 1H), 8.27 (d, J = 5.4 Hz, 1H), 7.91 (s, 1H), 7.63 (dd, J = 5.3, 1.4 Hz, 1H), 7.60 (d, J = 15.7 Hz, 1H), 7.13 (s, 1H), 7.10 (s, 1H), 6.84 (s, 1H), 3.41 (dd, J = 6.8, 4.5 Hz, 2H), 2.85 (t, J = 6.8 Hz, 2H).

[0476] Test method: Test method B was used. Retention time = 3.4 minutes.

[0477] Synthesis of Example 244

[0478]

[0479] Step 1: At -20 °C, morpholine (0.27 g, 3.15 mmol) in DCM (2 mL) was added dropwise to a solution of 2,4-dichloro-5-(iodomethyl)pyrimidine (1.00 g, 3.50 mmol) and DIEA (1.36 g, 10.50 mmol) in DCM (10 mL). The reaction mixture was stirred at -20 °C for 3 hours. The reaction mixture was extracted with DCM (30 mL), washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by flash column chromatography on silica gel (EA:DCM = 1 / 1) to give 4-((2,4-dichloropyrimidin-5-yl)methyl)morpholine (0.29 g, yield 32%). LCMS (m / z): [M+H]+ calculated 248.03; found 248.1.

[0480] Step 2: At room temperature, NaHCO3 (53 mg, 0.63 mmol) was added to a solution of 4-((2,4-dichloropyrimidin-5-yl)methyl)morpholine (130 mg, 0.52 mmol) and 3 (240 mg, 1.05 mmol) in DMF (3 mL). The reaction mixture was stirred at 90 °C for 2 hours. The reaction mixture was extracted with EA (20 mL x 3), washed with brine (10 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative HPLC to give Example 244 (4.12 mg, yield 1.7%). LCMS (m / z): [M+H]+ calculated 441.14; found 441.1. 11H NMR (400 MHz, CD3OD) δ 8.61 (s, 1H), 8.24 (d, J = 5.5 Hz, 1H), 7.54 (dd, J = 5.5, 1.6 Hz, 1H), 7.38 (d, J = 1.3 Hz, 1H), 7.10 (s, 1H), 3.70 - 3.67 (m, 6H), 3.56 (t, J = 7.0 Hz, 2H), 2.93 (t, J = 7.0 Hz, 2H), 2.57–2.54 (m, 4H).

[0481] Test method: Test method E was used. Retention time = 10 minutes.

[0482] Synthesis of Example 245

[0483]

[0484] Step 1: At -78 °C under nitrogen, n-BuLi (2.5 M, 11.6 mL, 1.0 equivalent) was added to a solution of 1,3-oxazole (2 g, 0.029 mol, 1.0 equivalent) in THF (30 mL). The reaction mixture was stirred at -78 °C for 1 hour. Then triisopropyl(trifluoromethanesulfonyloxy)silane (8.89 g, 0.029 mol, 1.0 equivalent) was added to the reaction mixture. The reaction mixture was stirred at -78 to 25 °C for 16 hours. Then it was quenched with NH4Cl solution and extracted with ethyl acetate (100 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 1) to give 2-(triisopropylsilyl)oxazole (5.0 g, yield 73%). LCMS (m / z): [M+H]+ calculated 226.15; found 226.2.

[0485] Step 2: At -78 °C under nitrogen, n-BuLi (2.5 M, 4.3 mL, 1.2 equivalents) was added to a solution of 2-(triisopropylsilyl)oxazole (2 g, 8.9 mmol, 1.0 equivalent) in THF (20 mL). The reaction mixture was stirred at -78 °C for 1 hour. Then triisopropyl borate (1.96 g, 0.0104 mol, 1.2 equivalents) was added to the reaction mixture and stirred at -78 °C for 2 hours. The reaction mixture was quenched with aqueous NH4Cl solution and extracted with ethyl acetate (80 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by HPLC on silica gel (acetonitrile:H2O = 10:1) to give (2-(triisopropylsilyl)oxazol-5-yl)boronic acid (350 mg, yield 14%). LCMS (m / z): [M+H]+ calculated 270.16; found 270.2.

[0486] Step 3: At room temperature under a nitrogen atmosphere with a balloon, add (2-(triisopropylsilyl)oxazol-5-yl)boronic acid (86 mg, 0.32 mmol, 1.5 equiv), K3PO4 (136 mg, 0.64 mmol, 3.0 equiv), Pd(dppf)Cl2 (16 mg, 0.021 mmol, 0.1 equiv) to a solution of Int-0013 (100 mg, 0.21 mmol, 1.0 equiv) in dioxane / H2O (10 mL). Then heat to 50 °C for 2 hours and then filter. Concentrate the filtrate in vacuo and purify the residue by flash column chromatography on silica gel (dichloromethane:methanol = 10 / 1) to obtain 2-(2-((2-chloro-5-(2-(triisopropylsilyl)oxazol-5-yl)pyrimidin-4-yl)oxy)pyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (55 mg, yield 41%). LCMS (m / z): [M+H]+ calculated 565.21; found 565.3.

[0487] Step 4: Add CsF (27 mg, 0.18 mmol, 2.0 equiv) to a solution of 2-(2-((2-chloro-5-(2-(triisopropylsilyl)oxazol-5-yl)pyrimidin-4-yl)oxy)pyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (50 mg, 0.089 mmol, 1.0 equiv) in DMF (2 mL). Stir the reaction mixture at 25 °C for 2 hours and then filter. Concentrate the filtrate in vacuo and purify the residue by HPLC on silica gel (acetonitrile:H2O (0.1% FA) = 30 - 70%) to obtain Example 245 as a yellow solid (5.96 mg, yield 16%). LCMS (m / z): [M+H]+ calculated 409.07; found 408.8. 1 1H NMR (400 MHz, d6-DMSO) δ 12.03 (s, 1H), 9.10 (s, 1H), 8.71 (s, 1H), 8.32 (d, J = 5.4 Hz, 1H), 7.82 (s, 1H), 7.68 - 7.67 (m, 1H), 7.62 (s, 1H), 7.13 - 7.12 (s, 2H), 3.47 - 3.46 (m, 2H), 2.87 - 2.83 (m, 2H).

[0488] Test method: Test method C used. Retention time = 1.04 minutes.

[0489] Synthesis of Example 246

[0490]

[0491] Step 1: To a solution of Int-0013 (80 mg, 0.17 mmol, 1 equiv) and (3-fluorophenyl)boronic acid (24 mg, 0.17 mmol, 1.0 equiv) in dioxane (5 mL) at room temperature were added purified water (1 mL), Xantphos Pd G3 (16 mg, 0.017 mmol), and K2CO3 (71 mg, 0.51 mmol). The mixture was stirred at 65 °C for 4 h under a nitrogen atmosphere, then filtered and the filtrate was concentrated. The residue was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm, 50 min linear gradient from 10% to 90% ACN) to give Example 246 (6.86 mg, 7.69%). LCMS (m / z): [M+H]+ calculated 435.09; found 436.2. 1 1H NMR (400 MHz, d6-DMSO) δ 12.06 (s, 1H), 8.86 (s, 1H), 8.45 (s, 1H), 8.23 (d, J = 5.4 Hz, 1H), 7.58 (dd, J = 6.4, 4.9 Hz, 2H), 7.56 - 7.54 (m, 3H), 7.37–7.23 (m, 1H), 7.07 (d, J = 3.4 Hz, 2H), 3.40 - 3.35 (m, 2H), 2.81 (t, J = 2.8 Hz, 2H).

[0492] Test method: Test method B was used. Retention time = 4.9 minutes.

[0493] Synthesis of Example 247

[0494]

[0495] Step 1: To a solution of Int-0013 (70 mg, 0.15 mmol) in dioxane (3 mL) and water (0.3 mL) were added K2CO3 (41 mg, 0.30 mmol), 2-trifluoromethyl-5-pyridineboronic acid (35 mg, 0.18 mmol), and tetrakis(triphenylphosphine)palladium (17 mg, 0.015 mmol). The mixture was stirred at 65 °C for 8 h under a nitrogen atmosphere. The mixture was filtered and the filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Samples were injected in DMF, 12.5 min linear gradient from 35% to 45% ACN) to give Example 247 (8.37 mg, 11%). LCMS (m / z): [M+H]+ calculated 487.08; found 487.1. 11H NMR (400 MHz, d6-DMSO) δ 12.02 (s, 1H), 9.13 (s, 1H), 9.03 (s, 1H), 8.47 (dd, J = 8.2, 1.9 Hz, 1H), 8.29 (d, J = 5.4 Hz, 1H), 8.12 (d, J = 8.2 Hz, 1H), 7.65–7.62 (m, 1H), 7.61 (s, 1H), 7.12 (s, 2H), 7.11 (d, J = 2.3 Hz, 0H), 3.41 (dd, J = 6.7, 4.4 Hz, 2H), 2.85 (t, J = 6.9 Hz, 2H).

[0496] Test method: Test method B was used. Retention time = 5.137 minutes.

[0497] Synthesis of Example 248

[0498]

[0499] Step 1: A solution of isopropylamine (111 mg, 1.89 mmol) and triethylamine (263 mg, 2.60 mmol) in DCM (2 mL) was added dropwise to a stirred suspension of 2,4-dichloropyrimidine-5-carbonyl chloride (500 mg, 2.365 mmol) in DCM (5 mL) at -60 °C under nitrogen. The reaction mixture was stirred at -60 °C for 3 hours. The reaction mixture was quenched with water (5 mL) and then extracted with DCM (5 mL x 3). The organic layer was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (DCM:EA = 25 / 1) to give 2,4-dichloro-N-isopropylpyrimidine-5-carboxamide as a white solid (350 mg, yield 50%). LCMS (m / z): [M+H]+ calculated 234.01; found 234.0.

[0500] Step 2: A solution of 2,4-dichloro-N-isopropylpyrimidine-5-carboxamide (100 mg, 0.43 mmol), 2-(2-hydroxypyridin-4-yl)-1H,5H,6H,7H-pyrrolo[3,2-c]pyridin-4-one (117 mg, 0.512 mmol) and NaHCO3 (179 mg, 2.14 mmol) in DMF (2 mL) was stirred at 90 °C for 1 hour under nitrogen and then filtered. The filtrate was concentrated in vacuo and the residue was purified by preparative HPLC to give Example 248 (10.29 mg, yield 5.31%). LCMS (m / z): [M+H]+ calculated 427.12; found 427.1. 1H NMR (400MHz, d6-DMSO) δ11.97(s,1H),8.79(s,1H),8.41(d,J=7.6Hz,1H),8.20(d,J=5.4Hz,1H),7.58(dd,J=5.4,1.4Hz,1H ),7.46(d,J=0.8Hz,1H),7.06(ss,2H),4.01–3.88(m,1H),3.37(dd,J=6.7,4.9Hz,2H),2.81(t,J=6.8Hz,2H),1.09(d,6H).

[0501] Test Method: Test Method F was used.

[0502] Synthesis of Example 249

[0503]

[0504] Step 1: To a solution of tert-butyl 2,4-dioxopiperidin-1-carboxylate (3 g, 0.014 mol) and 1,4-dibromobutane (4.53 g, 0.021 mol) in THF (50 mL) was added LiHMDS solution (1 M in THF, 84 mL, 0.084 mol) at -20 °C under a nitrogen atmosphere with a balloon, and then warmed to room temperature for 2 hours. The reaction mixture was quenched with NH4Cl solution and extracted with ethyl acetate. The combined organic layers were concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate: petroleum ether = 1 / 1) to give tert-butyl 8,10-dioxo-7-azaspiro[4.5]decane-7-carboxylate (0.8 g, yield 19%). LCMS (m / z): [M-55]+ calculated 212.1; found 212.2.

[0505] Step 2: To a solution of tert-butyl 8,10-dioxo-7-azaspiro[4.5]decane-7-carboxylate (440 mg, 1.64 mmol) and 2-bromo-1-(2-chloropyridin-4-yl)ethane-1-one (461 mg, 1.97 mmol) in ethanol / AcOH (12 mL, V / V=5 / 1) was added NH4OAc (1.26 g, 16.4 mmol) at room temperature under a nitrogen atmosphere with a balloon, and then the temperature was raised to 90°C and stirred for 2 hours. The mixture was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate: petroleum ether = 1 / 1) to give tert-butyl 2'-(2-chloropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (300 mg, yield 45%). LCMS (m / z): [M-55]+ Calculated 346.09; Found 346.2.

[0506] Step 3: To a solution of tert-butyl 2'-(2-chloropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (300 mg, 0.74 mmol) in AcOH (12 mL) was added water (4 mL). The mixture was stirred at 130 ° C for 3 days. The mixture was concentrated in vacuo and the residue was purified by C18 reverse phase column chromatography (H2O:ACN=4 / 1) to give 2'-(2-hydroxypyridin-4-yl)-5',6'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-4'(1'H)-one (200 mg, yield 85%). LCMS (m / z): [M+H]+ calculated 284.1; found 284.2.

[0507] Step 4: To a solution of 2'-(2-hydroxypyridin-4-yl)-5',6'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridin]-4'(1'H)-one (150 mg, 0.53 mmol) and 2,4-dichloro-5-iodopyrimidine (175 mg, 0.764 mmol) in DMF (5 mL) was added NaHCO3 (133 mg, 1.59 mmol). The mixture was stirred at 90° C. for 2 hours and then filtered. The filtrate was purified by C18 reverse phase column chromatography (H2O:ACN=4 / 1) to give 2'-(2-((2-chloro-5-iodopyrimidin-4-yl)oxy)pyridin-4-yl)-5',6'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-4'(1'H)-one (100 mg, yield 32%). LCMS (m / z): [M+H]+ calculated 522.0; found 522.1.

[0508] Step 5: To a solution of 2'-(2-((2-chloro-5-iodopyrimidin-4-yl)oxy)pyridin-4-yl)-5',6'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-4'(1'H)-one (100 mg, 0.019 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (45 mg, 0.23 mmol) in 1,4-dioxane / H2O (10 mL, V / V = 4 / 1) was added [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (14 mg, 0.0019 mmol) and tripotassium phosphate (81 mg, 0.3834 mmol). The reaction mixture was stirred at 65 °C under nitrogen for 2 h and then filtered. The filtrate was concentrated in vacuo and the residue was purified by preparative HPLC to give Example 249 (3.57 mg, yield 3.76%). LCMS (m / z): [M+H]+ calcd 463.1; found 463.1. 1 1H NMR (400 MHz, DMSO-d6) δ 11.48 (s, 1H), 9.59 (s, 1H), 9.33 (s, 1H), 9.16 (s, 1H), 8.28 (d, J = 5.4 Hz, 1H), 7.73 (dd, J = 5.4, 1.3 Hz, 1H), 7.67 (s, 1H), 7.16 (s, 1H), 7.06 (d, J = 2.2 Hz, 1H), 3.12 (d, J = 1.9 Hz, 2H), 1.86–1.63 (m, 8H).

[0509] Test method: Test method B was used.

[0510] Synthesis of Example 250

[0511]

[0512] Step 1: To a solution of Int-0013 (80 mg, 0.17 mmol) in dioxane (4 mL) and water (0.4 mL) were added K2CO3 (47 mg, 0.34 mmol), 3-methylisoxazole-4-boronic acid pinacol ester (40 mg, 0.19 mmol), and Pd(PPh3)4 (20 mg, 0.017 mmol). The mixture was stirred at 50 °C for 2 h under a nitrogen atmosphere. The mixture was filtered and the filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMF was injected, with a linear gradient from 30% to 40% ACN in 12.5 min), to give Example 250 as a white solid (25.42 mg, 35%). LCMS (m / z): [M+H]+ calculated 423.1; found 423.2. 1 H NMR (400 MHz, d6-DMSO) δ 12.00 (s, 1H), 9.25 (s, 1H), 8.85 (s, 1H), 8.29 (d, J = 5.4 Hz, 1H), 7.65 - 7.62 (m, 1H), 7.56 (s, 1H), 7.12 (d, J = 2.2 Hz, 2H), 3.41 (td, J = 6.7, 2.1 Hz, 2H), 2.85 (t, J = 6.8 Hz, 2H), 2.41 (s, 3H).

[0513] Test method: Test method B was used. Retention time = 3.861 min.

[0514] Synthesis of Example 251

[0515]

[0516] Step 1: To a solution of 5-bromo-2,4-dichloropyrimidine (1.0 g, 4.39 mmol, 1.0 equiv) in MeOH (1.5 mL) and THF (20 mL) at 0 °C was added CH3ONa (593 mg, 10.97 mmol, 2.5 equiv), and the reaction mixture was stirred at room temperature overnight. The mixture was diluted with water (10 mL) and extracted with EtOAc (30 mL × 3). The combined organic phases were washed with brine (30 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (20:1), to give 5-bromo-2,4-dimethoxypyrimidine (920 mg, 96%). LCMS-ESI (m / z): [M+H]+ calculated 218.97; found 218.87.

[0517] Step 2: A mixture of 5-bromo-2,4-dimethoxypyrimidine 2 (510 mg, 2.33 mmol, 1.0 equiv), tributyl(oxazol-2-yl)stannane (1.0 g, 2.79 mmol, 1.2 equiv) and Pd(PPh3)2Cl2 (82 mg, 116.42 μmol, 0.05 equiv) in DMF (4 mL) was stirred overnight at 90 °C under nitrogen. The mixture was diluted with water (30 mL) and extracted with EtOAc (20 ml×3). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (20:1) to give 2-(2,4-dimethoxypyrimidin-5-yl)oxazole (350 mg, 73%). LCMS-ESI (m / z): [M+H]+ calculated 208.06; found 208.14.

[0518] Step 3: NaI (759.63 mg, 5.07 mmol, 3.0 equiv) was added to a solution of 2-(2,4-dimethoxypyrimidin-5-yl)oxazole (350 mg, 1.69 mmol, 1.0 equiv) in AcOH (6 mL), and the reaction mixture was stirred at 110 °C for 1 h. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography, eluting with MeOH and DCM (1:20) to give 5-oxazol-2-ylpyrimidine-2,4-diol (250 mg, 83%). LCMS-ESI (m / z): [M+H]+ calculated 180.03; found 180.13.

[0519] Step 4: DIEA (180 mg, 1.40 mmol, 243.09 μL, 1.0 equiv) was added to a solution of 5-oxazol-2-ylpyrimidine-2,4-diol (250 mg, 1.40 mmol, 1.0 equiv) in POCl3 (5 mL), and the reaction was stirred at 110 °C for 2 h. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (20:1) to give 2-(2,4-dichloropyrimidin-5-yl)oxazole (90 mg, 30%). LCMS-ESI (m / z): [M+H]+ calculated 215.97; found 216.07.

[0520] Step 5: To a solution of Int-0012 (90 mg, 392.6 μmol, 1.0 eq) in DMF (1 mL) was added 2-(2,4-dichloropyrimidin-5-yl)oxazole (85 mg, 392.6 μmol, 1.0 eq) and NaHCO3 (66 mg, 785.2 μmol, 2.0 eq), and the reaction mixture was stirred at 100 °C for 0.5 h. The reaction mixture was purified by preparative HPLC to give Example 251 (8.4 mg, 5%). LCMS-ESI (m / z): [M+H]+ calculated 409.07; found 408.93. 1 1H NMR (DMSO-d6): δ 12.02 (s, 1H), 9.62 (s, 1H), 8.43 - 8.42 (m, 1H), 8.28 - 8.27 (m, 1H), 7.65 - 7.64 (m, 1H), 7.59 (s, 1H), 7.55 (s, 1H), 7.13 (s, 2H), 3.44 - 3.40 (m, 2H), 2.87 - 2.84 (m, 2H).

[0521] Test method: Test method A used. Retention time = 2.04 min.

[0522] Synthesis of Example 252

[0523]

[0524] Step 1: To a solution of (2,4-dimethoxypyrimidin-5-yl)boronic acid (1.39 g, 7.55 mmol, 1.2 eq) in 1,4-dioxane (50 mL) and water (5 mL) was added 2-bromopyrimidine (1.0 g, 6.29 mmol, 1.0 eq), Pd(dppf)Cl2·DCM (103 mg, 125.80 μmol, 0.02 eq) and NaHCO3 (1.59 g, 18.87 mmol, 3.0 eq), and the reaction was stirred at 90 °C for 16 h. The reaction mixture was concentrated, and the residue was purified by silica gel column (eluted with DCM solution of 0 - 5% CH3OH) to give 2',4'-dimethoxy-2,5'-bipyrimidine (830 mg, 61%). LCMS-ESI (m / z): [M+H]+ calculated 219.22; found 219.00.

[0525] Step 2: A solution of 2',4'-dimethoxy-2,5'-bipyrimidine (328 mg, 1.50 mmol, 1.0 equiv) and NaI (450 mg, 3.01 mmol, 2.0 equiv) in AcOH (6 mL) was stirred at 110 °C for 1 h. The reaction solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (eluting with 0 - 10% MeOH in DCM) to give [2,5'-bipyrimidine]-2',4'-diol (267 mg, 93%). LCMS-ESI (m / z): [M+H]+ calculated 191.16; found 190.76.

[0526] Step 3: To a solution of [2,5'-bipyrimidine]-2',4'-diol (267 mg, 1.40 mmol, 1.0 equiv) in POCl3 (4 mL) was added DIEA (181 mg, 1.40 mmol, 1.0 equiv), and the reaction was stirred at 110 °C for 3 h. The reaction solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (eluting with a PE solution of 0 - 20% EA) to give 2',4'-dichloro-2,5'-bipyrimidine (150 mg, 47%). LCMS-ESI (m / z): [M+H]+ calculated 227.05; found 226.91.

[0527] Step 4: To a solution of 2',4'-dichloro-2,5'-bipyrimidine (110 mg, 1.40 mmol, 1.0 equiv) in DMAc (10 mL) were added Int-0012 (122 mg, 532.92 μmol, 1.1 equiv) and t-BuOK (82 mg, 726.71 μmol, 1.5 equiv), and the reaction was stirred at 60 °C for 2 h. The residue was purified by preparative HPLC to give Example 252 (2.1 mg, 1%). LCMS-ESI (m / z): [M+H]+ calculated 419.83; found 419.92. 1 1H NMR (400 MHz, DMSO-d6): δ 11.99 (s, 1H), 9.24 (s, 1H), 8.99 - 8.97 (m, 1H), 8.91 (s, 1H), 8.18 (d, J = 4.0 Hz, 1H), 7.59 - 7.53 (m, 2H), 7.52 (s, 1H), 7.12 (s, 2H), 3.43 - 3.40 (m, 2H), 2.85 (t, J = 6.0 Hz, 2H).

[0528] Test method: Test method A was used. Retention time = 2.02 min.

[0529] Synthesis of Example 253

[0530]

[0531] Step 1: A solution of 2,4-dichloro-5-pyrimidinecarbonyl chloride (2 g, 9.5 mmol) in t-BuOH (20 mL) was stirred at 30 °C for 16 h under a nitrogen atmosphere. When TLC analysis showed the reaction was complete, the reaction mixture was diluted with EtOAc (30 mL), and then washed with water (50 mL), saturated aqueous NaHCO3 (40 mL), and brine (40 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluted with petroleum ether / EtOAc = 10:1) to give tert-butyl 2,4-dichloropyrimidine-5-carboxylate as a colorless oil (800 mg, purity 90%, yield 30%). LCMS (m / z): [M+H]+ calculated 249.0; found 249.1.

[0532] Step 2: To a solution of tert-butyl 2,4-dichloropyrimidine-5-carboxylate (305 mg, 1.22 mmol) in DMF (5 mL) was added Int-0012 (300 mg, 1.11 mmol) and K2CO3 (307 mg, 2.22 mmol). The mixture was stirred at 50 °C for 2 h under a nitrogen atmosphere. The mixture was filtered, and the filtrate was purified by preparative HPLC using a gradient of H2O (0.1% FA) / ACN (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMF was injected, and the linear gradient was from 35% to 60% ACN in 12.5 min), to give Example 253 (6.36 mg, purity 99.4%, yield 1.29%). LCMS (m / z): [M+H]+ calculated 442.1; found 442.2. 1 H NMR (400 MHz, d6-DMSO) δ 12.02 (s, 1H), 9.09 (s, 1H), 8.19 (d, J = 5.4 Hz, 1H), 7.60 (dd, J = 5.4, 1.4 Hz, 1H), 7.52 (s, 1H), 7.14 (d, J = 2.4 Hz, 1H), 7.12 (s, 1H), 3.41 (td, J = 6.9, 2.4 Hz, 2H), 2.85 (t, J = 6.8 Hz, 2H), 1.45 (s, 9H).

[0533] Test method: Test method B was used. Retention time = 4.975 min.

[0534] Synthesis of Example 254

[0535]

[0536] Step 1: At 20 °C under nitrogen, pyrrolidine (220 mg, 3.1 mmol) and DIEA (900 mg, 7.0 mmol) were added to a solution of 2,4-dichloro-5-(iodomethyl)pyrimidine (1.0 g, 3.5 mmol) in DCM (10 mL). The reaction mixture was stirred at -20 °C for 2 h. The mixture was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (PE / EA = 4 / 1) to give 2,4-dichloro-5-(pyrrolidin-1-ylmethyl)pyrimidine (300 mg, yield 37%). LCMS (m / z): [M+H]+ calculated 232.0; found 232.1.

[0537] Step 2: At 50 °C under nitrogen, NaHCO3 (163 mg, 1.9 mmol) and Int-0012 (222 mg, 0.78 mmol) were added to a solution of 2,4-dichloro-5-(pyrrolidin-1-ylmethyl)pyrimidine (150 mg, 0.65 mmol) in DMF (1.5 mL). The reaction mixture was stirred at 50 °C for 2 h. Then it was filtered and the filtrate was purified by preparative HPLC using a gradient of H2O (0.1% FA) / ACN (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMF was injected and the linear gradient was from 10% to 10% ACN in 10.5 min), to give Example 254 (14.08 mg, 5%). LCMS (m / z): [M+H]+ calculated 425.1; found 425.2. 1 1H NMR (400 MHz, d6-DMSO) δ 12.01 (s, 1H), 8.67 (s, 1H), 8.26 (d, J = 5.4 Hz, 1H), 8.19 (s, 1H), 7.62 (dd, J = 5.4, 1.3 Hz, 1H), 7.50 (s, 1H), 7.12 (d, J = 2.2 Hz, 1H), 7.10 (s, 1H), 3.71 (s, 2H), 3.43–3.40 (m, 2H), 2.85 (t, J = 6.8 Hz, 2H), 2.55 (br s, 4H), 1.72-1.71 (m, 4H).

[0538] Test method: Test method B was used. Retention time = 3.305 min.

[0539] Synthesis of Example 255

[0540]

[0541] Step 1: At -20 °C under nitrogen, add a solution of piperidine (147 mg, 1.73 mmol, 1.0 equiv) and DIEA (447 mg, 3.46 mmol, 2.0 equiv) in DCM (3 mL) to a solution of 2,4-dichloro-5-(iodomethyl)pyrimidine (500 mg, 1.73 mmol, 1.0 equiv) in DCM (5 mL). Stir the reaction mixture at -20 °C for 1 hour. Concentrate the reaction mixture in vacuo and purify the residue by flash column chromatography on silica gel (EtOAc: petroleum ether = 1 / 1) to give the product, 2,4-dichloro-5-(piperidin-1-ylmethyl)pyrimidine (130 mg, yield 27%). LCMS (m / z): [M+H]+ calculated 246.1; found 245.9.

[0542] Step 2: To a solution of 2,4-dichloro-5-(piperidin-1-ylmethyl)pyrimidine (100 mg, 0.41 mmol, 1.0 equiv) in DMF (5 mL), add Int-0014 (100 mg, 0.41 mmol, 1.0 equiv), NaHCO3 (102 mg, 1.22 mmol, 3.0 equiv). Stir the reaction mixture at 90 °C for 2 hours. Filter the mixture and purify the filtrate by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Inject the sample in DMF, 50-minute linear gradient from 10% to 90% ACN) to give Example 255 (5.61 mg, 1.53%). LCMS (m / z): [M+H]+ calculated 457.2; found 457.2. 1 1H NMR (400 MHz, d6-DMSO) δ 11.99 (s, 1H), 9.75 (s, 1H), 8.88 (s, 1H), 8.42 (d, J = 2.8 Hz, 1H), 7.72 (d, J = 5.4 Hz, 1H), 7.20 (s, 1H), 7.06 (s, 1H), 4.46 (s, 2H), 3.51 - 3.50 (m, 2H), 3.45 - 3.42 (m, 2H), 3.11 - 3.05 (m, 2H), 2.88 (t, J = 6.8 Hz, 2H), 1.85 - 1.80 (m, 2H), 1.75 - 1.60 (m, 3H), 1.45 - 1.35 (m, 1H).

[0543] Test method: Test method B used. Retention time = 3.44 minutes.

[0544] Synthesis of Example 256

[0545]

[0546] Step 1: Add a solution of Int-007 (10 mg, 0.04 mmol, 1.25 equiv) in ACN (1 mL) to a vial. Add 2,6-dichloropyrimidine-4-carbonitrile (5.6 mg, 0.03 mmol, 1.0 equiv) and K2CO3 (18 mg, 0.13 mmol, 4.0 equiv) to the solution. Heat the reaction mixture to 60 °C and stir for 2 h. Cool the solution to room temperature, dilute with EtOAc, wash with brine, dry over MgSO4, and concentrate in vacuo. Purify the crude product by preparative HPLC, H2O (+0.1% FA) / CAN, gradient 5 - 95%, to give Example 256 (3 mg, 24% yield). LCMS (m / z): [M+H]+ calcd 384.0; found 385.0. 1 1H NMR (400 MHz, d6-DMSO) δ 12.00 (s, 1H), 8.90 (s, 1H), 7.22 (s, 1H), 7.12 (s, 1H), 7.01 - 6.94 (m, 1H), 3.43 (dt, J = 7.1, 2.4 Hz, 2H), 2.90 (t, J = 5.9 Hz, 2H).

[0547] Synthesis of Example 257

[0548]

[0549] Step 1: Add ethyl 4,4-difluoro-3-oxobutanoate (1.00 g, 6.02 mmol) and urea (0.723 g, 12.04 mmol, 2.0 equiv) to a flask. Flush the flask with Ar by pumping, then suspend the solid in PhMe (30 mL, 0.2 M). Treat the suspension with NaOEt (0.819 g, 12.04 mmol, 2.0 equiv), heat to 130 °C and stir for 48 h. Cool the solution to room temperature, concentrate, then triturate with EtOAc overnight. Then filter the solid and dry in vacuo to give 6-(difluoromethyl)pyrimidine-2,4-diol as a dark solid (0.975 g, quantitative). LCMS (m / z): [M+H]+ calcd 163.0; found 163.0.

[0550] Step 2: 6-(Difluoromethyl)pyrimidine-2,4-diol (0.975 g, 6.02 mmol) was added to a flask and dissolved in CAN (10 mL, 0.6 M). The reaction solution was cooled to 0 °C and treated with POCl3 (4.06 g, 26.47 mmol, 4.4 eq) and DIPEA (0.660 g, 5.11 mmol, 0.85 eq). The reaction solution was then warmed to 95 °C and stirred overnight. Then, the solution was cooled to room temperature, quenched with ice water, extracted with EtOAc, washed with one volume of brine, dried over MgSO4 and concentrated. The crude mixture was purified by normal phase, 0-100% EtOAc / heptane, to give 2,4-dichloro-6-(difluoromethyl)pyrimidine (0.148 g, 12.4%). 1 1H NMR (400 MHz, CDCl3) δ 7.54 (s, 1H), 6.46 (br t, J = 52.4, 1H).

[0551] Step 3: Int-007 (0.020 g, 0.08 mmol), 2,4-dichloro-6-(difluoromethyl)pyrimidine (0.016 g, 0.08 mmol, 1.0 eq), K2CO3 (0.045 g, 0.32 mmol, 4.0 eq) were added to a flask, dissolved in ACN (1 mL, 0.08 M) and warmed to 80 °C for 2 h. The reaction solution was cooled to room temperature, diluted with EtOAc, washed with one volume of brine, dried over MgSO4 and concentrated. The mixture was purified by preparative HPLC, 5-95% ACN / H2O + 0.05% formic acid, to give Example 257 (0.005 g, 15.1%). LCMS (m / z): [M+H]+ calculated 410.6; found 410.0. 1 1H NMR (400 MHz, d6-DMSO) δ 8.59 (s, 1H), 8.06 - 8.02 (m, 1H), 7.21 (br s, 1H), 7.16 (s, 1H), 7.11 (s, 1H), 6.95 (t, J = 6.9 Hz, 1H), 3.43 (dt, J = 7.0, 2.5 Hz, 2H), 2.90 (t, J = 6.6 Hz, 2H).

[0552] Synthesis of Example 258

[0553]

[0554] Step 1: At 0 °C, dimethylamine (355 mg, 7.89 mmol, 1 equiv) and triethylamine (798 mg, 7.89 mmol, 1 equiv) were added to a solution of 2,4-dichloropyrimidine-5-carbonyl chloride (2.00 g, 9.46 mmol, 1.2 equiv) in DCM (20 mL). The reaction mixture was stirred at 25 °C for 1 h under nitrogen. After adding water, the mixture was extracted with DCM (10 mL × 2). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by a C18 reverse-phase column (H2O:ACN = 1 / 1) to give the product 2,4-dichloro-N,N-dimethylpyrimidine-5-carboxamide (140 mg, yield 5%). LCMS (m / z): [M+H]+ calculated 220.00; found 220.0.

[0555] Step 2: A suspension of 2,4-dichloro-N,N-dimethylpyrimidine-5-carboxamide (140 mg, 0.64 mmol, 1 equiv), Int-0012 (175 mg, 0.76 mmol, 1.2 equiv) and sodium bicarbonate (267 mg, 3.18 mmol, 1.5 equiv) in DMF (2 mL) was stirred at 90 °C for 1 h under nitrogen and then filtered. The filtrate was purified by preparative HPLC to give the product Example 258 (1.30 mg, yield 0.44%). LCMS (m / z): [M+H]+ calculated 413.11; found 413.1. 1 1H NMR (400 MHz, d6-DMSO δ 12.00 (s, 1H), 8.79 (s, 1H), 8.27 (d, 1H), 7.64 (d, J = 5.4 Hz, 1H), 7.55 (s, 1H), 7.15 (d, J = 2.2 Hz, 1H), 7.11 (s, 1H), 3.41 (td, J = 6.5, 2.4 Hz, 2H), 3.00 (d, J = 6.1 Hz, 6H), 2.85 (t, J = 6.5 Hz, 2H).

[0556] Test method: Test method F was used.

[0557] Synthesis of Example 259

[0558]

[0559] Step 1: To a mixture of 2,3-dichloro-N-methoxy-N-methylisonicotinamide (5.0 g, 0.026 mol, 1.0 equiv) in DCM (100 mL) was added HATU (14.83 g, 0.039 mol, 1.5 equiv), DIEA (13.44 g, 0.10 mol, 4 equiv), and then N,O-dimethylhydroxylamine hydrochloride (3.82 g, 0.039 mol, 1.5 equiv). The reaction mixture was stirred at 25 °C for 16 h. Then it was diluted with water (100 mL) and extracted with DCM (200 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo to give 2,3-dichloro-N-methoxy-N-methylisonicotinamide (8.0 g, crude). LCMS (m / z): [M+H]+ calculated 235.06; found 235.1.

[0560] Step 2: To a mixture of 2,3-dichloro-N-methoxy-N-methylisonicotinamide (7.5 g, 0.032 mol, 1 equiv) in THF (100 mL) at 0 °C was slowly added CH3MgBr (1 M in THF, 36 mL, 0.036 mol, 1.1 equiv). The reaction mixture was stirred at 0 °C for 1 h. The reaction mixture was quenched with saturated aqueous ammonium chloride. Then it was extracted with ethyl acetate (200 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo to give the crude product. The crude product was purified by flash chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 2) to give 1-(2,3-dichloropyridin-4-yl)ethan-1-one (4.0 g, yield 59%). LCMS (m / z): [M+H]+ calculated 190.02; found 190.0.

[0561] Step 3: To a mixture of 1-(2,3-dichloropyridin-4-yl)ethan-1-one (2.0 g, 0.011 mol, 1.0 equiv) in HBr (30% in AcOH, 30 mL). Then Br2 (0.6 mL, 10.8 mmol, 1.0 equiv) was slowly added at 0 °C. The reaction mixture was stirred at 25 °C for 16 h. Methyl tert-butyl ether (30 mL) was added to the reaction solution. After filtration, the filter cake was collected to give 2-bromo-1-(2,3-dichloropyridin-4-yl)ethan-1-one as a white solid (2.0 g, yield 97%). LCMS (m / z): [M+H]+ 267.9; found 268.2.

[0562] Step 4: A mixture of 2-bromo-1-(2,3-dichloropyridin-4-yl)ethan-1-one (2.0 g, 0.01 mol, 1.0 equiv), piperidine-2,4-dione (1.67 g, 0.02 mol, 2.0 equiv), AcOH (0.64 g, 0.015 mol, 1.5 equiv), and NH4OAc (1.32 g, 0.02 mol, 2.0 equiv) in EtOH (30 mL) was stirred at 80 °C for 16 h. The reaction mixture was diluted with water (20 mL) to form a yellow precipitate, which was collected by filtration to give 2-(2,3-dichloropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (1.2 g, yield 60%). LCMS (m / z): [M+H]+ calculated 282.1; found 282.2.

[0563] Step 5: To a mixture of 2-(2,3-dichloropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (500 mg, 1.6 mmol, 1.0 equiv) in 1,4-dioxane (3 mL) was added Pd2(dba)3 (295 mg, 0.32 mmol, 0.2 equiv), t-BuXPhos (70 mg, 0.15 mmol, 0.1 equiv), KOH (360 mg, 6 mmol, 4 equiv), and H2O (1 mL). The reaction mixture was stirred at 100 °C for 1 h under microwave. The aqueous phase was lyophilized to give 2-(3-chloro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (200 mg, yield 45%). LCMS (m / z): [M+H]+ calculated 264.1; found 264.2.

[0564] Step 6: A mixture of 2-(3-chloro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (80 mg, 0.28 mmol, 1.0 equiv), 2,4-dichloro-5-(ethoxymethyl)pyrimidine (50 mg, 0.24 mmol, 0.8 equiv), and NaHCO3 (76 mg, 0.83 mmol, 3.0 equiv) in DMF (4 mL) was stirred at 90 °C for 2 h. The solid was filtered off, and the filtrate was purified by preparative HPLC (acetonitrile: H2O (0.1% FA) = 30–70%) to give Example 259 (5.28 mg, yield 4.03%). LCMS (m / z): [M+H]+ calculated 434.3; found 434.2. 11H NMR (400 MHz, d6-DMSO) δ 12.04 (s, 1H), 8.71 (s, 1H), 8.27 (d, J = 5.3 Hz, 1H), 7.65 (d, J = 5.4 Hz, 1H), 7.16 (d, J = 11.1 Hz, 2H), 4.59 (s, 2H), 3.55 (q, J = 7.0 Hz, 2H), 3.39 (td, J = 6.8, 2.3 Hz, 2H), 2.85 (t, J = 6.8 Hz, 2H), 1.14 (t, J = 7.0 Hz, 3H).

[0565] Test method: Test method B was used. Retention time was 4.61 minutes.

[0566] Synthesis of Example 260

[0567]

[0568] Step 1: At 0 °C, NaH (60%, 103 mg, 2.59 mmol, 1.5 equivalents) was added to a mixture of pyrrolidin-2-one (176 mg, 2.08 mmol, 1.2 equivalents) in THF (10 mL), and the resulting mixture was stirred for 10 minutes. Then, 2,4-dichloro-5-(iodomethyl)pyrimidine (500 mg, 1.73 mmol, 1.0 equivalent) was added at 0 °C. The resulting mixture was stirred at 25 °C for another 1 hour. Acetic acid (1 mL) was added to quench the reaction, and then the mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then concentrated. The residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 2) to obtain 1-((2,4-dichloropyrimidin-5-yl)methyl)pyrrolidin-2-one (100 mg, yield 21%). LCMS (m / z): [M+H]+ calculated 246.1; found 246.2.

[0569] Step 2: A mixture of 1-((2,4-dichloropyrimidin-5-yl)methyl)pyrrolidin-2-one (100 mg, 0.41 mmol, 1.0 equivalent), Int-007 (93 mg, 0.41 mmol, 1.0 equivalent), and NaHCO3 (68 mg, 0.81 mmol, 2.0 equivalents) in DMF (5 mL) was stirred at 90 °C for 2 hours. The solid was filtered out, and the filtrate was purified by preparative HPLC (acetonitrile:H2O (0.1% FA) = 30 - 70%) to obtain Example 260 (1.93 mg, yield 1%). LCMS (m / z): [M+H]+ calculated 457.1; found 457.2. 11H NMR (400 MHz, d6-DMSO) δ 12.16 (s, 1H), 8.65 (s, 1H), 8.36 (s, 1H), 8.18 (d, J = 5.3 Hz, 1H), 7.82 (t, J = 5.3 Hz, 1H), 7.20 (s, 1H), 6.99 (d, J = 2.8 Hz, 1H), 4.54 (s, 2H), 3.44–3.38 (m, 4H), 2.89 (t, J = 6.7 Hz, 2H), 2.26 (t, J = 8.0 Hz, 2H), 2.00–1.93 (m, 2H).

[0570] Test method: Test method B was used. Retention time = 3.92 minutes.

[0571] Synthesis of Example 261

[0572]

[0573] Step 1: 2-Bromo-5-methyl-1,3,4-thiadiazole 1 (1 g, 5.62 mmol, 1.0 equiv), (2,4-dimethoxypyrimidin-5-yl)boronic acid (1.24 g, 6.74 mmol, 1.2 equiv), cesium carbonate (3.7 g, 11.24 mmol, 2.0 equiv) and Pd(dppf)Cl2·CH2Cl2 (459 mg, 560 μmol, 0.1 equiv) in a mixture of water (4 mL) and 1,4-dioxane (40 mL) were stirred overnight at 90 °C. The mixture was diluted with EtOAc (50 mL) and washed with water (50 mL × 3) and brine (50 mL × 3). The organic layer was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (0 - 50% EtOAc in PE) to give 2-(2,4-dimethoxypyrimidin-5-yl)-5-methyl-1,3,4-thiadiazole (910 mg, 68%). LCMS-ESI (m / z): [M+H]+ calculated 239.05; found 239.11.

[0574] Step 2: To a solution of 2-(2,4-dimethoxypyrimidin-5-yl)-5-methyl-1,3,4-thiadiazole (500 mg, 2.1 mmol, 1.0 equiv) in acetic acid (10 mL) was added NaI (540 mg, 3.6 mmol, 3.0 equiv). The reaction was then stirred at 110 °C for 1 h. The reaction mixture was concentrated and the residue was purified by column chromatography (0 - 3% MeOH in DCM) to give 5-(5-methyl-1,3,4-thiadiazol-2-yl)pyrimidine-2,4-diol (400 mg, 90%). LCMS-ESI (m / z): [M+H]+ calculated 211.02; found 210.98.

[0575] Step 3: A few drops of DIEA were added to a solution of 5-(5-methyl-1,3,4-thiadiazol-2-yl)pyrimidine-2,4-diol (400 mg, 1.9 mmol, 1.0 equiv) in POCl3 (5 mL). The solution was stirred at 110 °C for 2 h, then the solution was added dropwise to cold aqueous NaHCO3 solution. The mixture was extracted with EtOAc (50 mL), washed with water (50 mL) and brine (50 mL). The organic layer was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (0 - 10% EtOAc in PE) to give 2-(2,4-dichloropyrimidin-5-yl)-5-methyl-1,3,4-thiadiazole (180 mg, 38%). LCMS-ESI (m / z): [M + H]+ calculated 246.95; found 247.01.

[0576] Step 4: A mixture of 2-(2,4-dichloropyrimidin-5-yl)-5-methyl-1,3,4-thiadiazole (80 mg, 330 μmol, 1.0 equiv), 2-(2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (76 mg, 330 μmol, 1.0 equiv) and NaHCO3 (28 mg, 330 μmol, 1.0 equiv) in DMAc (5 mL) was stirred at 60 °C for 2 h. The solution was purified by preparative HPLC to give Example 261 (2.2 mg, 2%). LCMS-ESI (m / z): [M + H]+ calculated 440.06; found 439.91. 1 1H NMR (400 MHz, DMSO-d6): δ 12.03 (s, 1H), 9.49 (s, 1H), 8.32 (s, d, J = 4.0 Hz, 1H), 7.69 (t, J = 4.0 Hz, 1H), 7.65 (d, J = 4.0 Hz, 1H), 7.13 (t, J = 4.0 Hz, 2H), 3.44 - 3.40 (m, 2H), 2.88 - 2.84 (m, 2H), 2.82 (s, 3H).

[0577] Test method: Test method A used. Retention time = 2.07 min.

[0578] Synthesis of Example 262

[0579]

[0580] Step 1: To a solution of Int-007 (500 mg, 2.02 mmol, 1.0 eq) in DMF (4 mL) was added 2,4-dichloro-5-(ethoxymethyl)pyrimidine (417 mg, 2.02 mmol, 1.0 eq) and NaHCO3 (510 mg, 6.07 mmol, 3.0 eq). The resulting mixture was stirred at 90 °C under nitrogen for 3 h. The solid was filtered off and the filtrate was purified directly by preparative HPLC (ACN-H2O, 0.1% FA) to give 2-(2-((4-chloro-5-(ethoxymethyl)pyrimidin-2-yl)oxy)-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (120.05 mg, 14.2%). LCMS (m / z): [M+H]+ calcd 418.1; found 418.0. 1 1H NMR (400 MHz, d6-DMSO) δ 12.03 (s, 1H), 8.71 (s, 1H), 8.15 (d, J = 5.3 Hz, 1H), 7.76 (t, J = 5.4 Hz, 1H), 7.18 (s, 1H), 6.97 (t, J = 2.7 Hz, 1H), 4.52 (s, 2H), 3.56 (q, J = 7.0 Hz, 2H), 3.42 (td, J = 6.8, 2.5 Hz, 2H), 2.88 (t, J = 6.8 Hz, 2H), 1.16 (t, J = 7.0 Hz, 3H).

[0581] Test method: Test method B was used.

[0582] Synthesis of Example 263

[0583]

[0584] Step 1: To a solution of 4-(tributylstannyl)-2-(triisopropylsilyl)-1,3-oxazole (330 mg, 0.64 mmol) in anhydrous ACN (8 mL) was added Int-0013 (100 mg, 0.21 mmol) and bis(triphenylphosphine)palladium(II) chloride (15 mg, 0.021 mmol). The mixture was stirred at 75 °C for 18 h. The residue was purified by flash chromatography (C-18 reverse phase column, water / ACN, 4:1) to give 2-(2-((2-chloro-5-(2-(triisopropylsilyl)oxazol-4-yl)pyrimidin-4-yl)oxy)pyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (30 mg, purity 85%, yield 21%). LCMS (m / z): [M+H]+ calcd 565.2; found 565.3.

[0585] Step 2: To a solution of 2-(2-((2-chloro-5-(2-(triisopropylsilyl)oxazol-4-yl)pyrimidin-4-yl)oxy)pyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (30 mg, 0.053 mmol) in DMF (5 mL) was added CsF (41 mg, 0.27 mmol). The mixture was stirred at 25 °C for 2 h under a nitrogen atmosphere. The mixture was filtered and the filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMF was injected, with a linear gradient from 25% to 60% ACN over 12.5 min), to give Example 263 (4.99 mg, purity 96.1%, yield 22%). LCMS (m / z): [M+H]+ calculated 409.1; found 409.2. 1 H NMR (400 MHz, d6-DMSO) δ 11.97 (s, 1H), 9.15 (s, 1H), 8.70 (d, J = 0.5 Hz, 1H), 8.65 (d, J = 0.7 Hz, 1H), 8.28 (d, J = 5.4 Hz, 1H), 7.63 (dd, J = 5.3, 1.4 Hz, 1H), 7.57 (s, 1H), 7.08 (d, J = 2.3 Hz, 1H), 7.07 (s, 1H), 3.37 (td, J = 6.8, 2.2 Hz, 2H), 2.80 (t, J = 6.9 Hz, 2H).

[0586] Test method: Test method B was used. Retention time = 4.657 min.

[0587] Synthesis of Example 264

[0588]

[0589] Step 1: At -78 °C, n-BuLi (2.4 M in THF, 21.9 mL, 52 mmol, 1.15 equivalents) was added to a solution of 5-bromo-2,4-dimethoxypyrimidine (10 g, 46.12 mmol, 1.0 equivalent) in THF (100 mL). The reaction mixture was stirred at -78 °C for 1 hour. Then DMF (4.01 g, 55.34 mmol, 1.2 equivalents) was added to the reaction. The reaction mixture was stirred at -78 °C for 1 hour. The reaction mixture was quenched with NH4Cl solution. The mixture was extracted with ethyl acetate (100 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 4) to give 2,4-dimethoxypyrimidine-5-carbaldehyde (5.8 g, yield 67%). LCMS (m / z): [M+H]+ calculated 169.2; found 169.1.

[0590] Step 2: To a solution of 2,4-dimethoxypyrimidine-5-carbaldehyde (5.5 g, 33.01 mmol, 1.0 equivalent) in EtOH (50 mL) was added NH2OH·HCl (3.41 g, 49.52 mmol, 1.5 equivalents) and TEA (4.96 g, 49.52 mmol, 1.5 equivalents). The reaction mixture was stirred at 25 °C for 3 hours and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 4) to give (E)-2,4-dimethoxypyrimidine-5-carbaldehyde oxime (3.3 g, yield 49%). LCMS (m / z): [M+H]+ calculated 184.2; found 184.1.

[0591] Step 3: At 0 °C, NCS (2.57 g, 19.49 mmol, 1.1 eq) was added to a solution of (E)-2,4-dimethoxypyrimidine-5-carbaldehyde oxime (3.2 g, 17.72 mmol, 1.0 eq) in DMF (20 mL). The reaction mixture was stirred at 0 °C for 2 h. Then, a solution of TMSA (5.16 g, 53.16 mmol, 3.0 eq) in THF (30 mL) and TEA (3.54 g, 35.44 mmol, 2.0 eq) were added to the reaction at 0 °C. The reaction mixture was stirred at 25 °C for 14 h. The reaction mixture was quenched with a NaHCO3 solution. The mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate: petroleum ether = 1 / 4) to give 3-(2,4-dimethoxypyrimidin-5-yl)-5-(trimethylsilyl)isoxazole (3.4 g, yield 59%). LCMS (m / z): [M+H]+ calculated 280.1; found 280.1.

[0592] Step 4: K2CO3 (2.53 g, 18.28 mmol, 1.5 eq) was added to a solution of 3-(2,4-dimethoxypyrimidin-5-yl)-5-(trimethylsilyl)isoxazole (3.4 g, 12.19 mmol, 1.0 eq) in MeOH (40 mL). The reaction mixture was stirred at 25 °C for 1 h. The mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo. The crude product was used in the next step without any further purification. The crude product (2.2 g, yield 81%) was obtained. LCMS (m / z): [M+H]+ calculated 208.06; found 208.2.

[0593] Step 5: At 0 °C, BBr3 (1 M in DCM, 35.6 mL, 35.6 mmol, 0.4 eq) was added to a solution of 3-(2,4-dimethoxypyrimidin-5-yl)isoxazole (1.85 g, 88.95 mmol, 1.0 eq) in DCM (20 mL). The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was quenched with a NaHCO3 solution. The mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by silica gel reverse phase column (acetonitrile: water = 1 / 9) to give 5-(isoxazol-3-yl)pyrimidine-2,4-diol (550 mg, yield 31%). LCMS (m / z): [M+H]+ calculated 180.03; found 180.1.

[0594] Step 6: A solution of 5-(isoxazol-3-yl)pyrimidine-2,4-diol (200 mg, 1.12 mmol, 1.0 equiv) in POCl3 / DIEA = 10:1 (11 mL) was stirred at 120 °C for 2 h. The reaction solution was concentrated in vacuo. The resulting mixture was washed with saturated aqueous sodium bicarbonate and extracted with ethyl acetate (10 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 3) to give 3-(2,4-dichloropyrimidin-5-yl)isoxazole (140 mg, yield 52%). LCMS (m / z): [M+H]+ calculated 215.97; found 215.9.

[0595] Step 7: To a solution of 3-(2,4-dichloropyrimidin-5-yl)isoxazole (20 mg, 0.093 mmol, 1.0 equiv) in DMF (2 mL) was added Int-007 (27 mg, 0.11 mmol, 1.2 equiv) and NaHCO3 (23 mg, 0.28 mmol, 3.0 equiv). The reaction mixture was stirred at 25 °C for 16 h and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by HPLC on silica gel (acetonitrile:H2O (0.1% FA) = 30–70%) to give Example 264 (7.28 mg, yield 18%). LCMS (m / z): [M+H]+ calculated 427.07; found 426.9. 1 1H NMR (400 MHz, d6-DMSO) δ 12.05 (s, 1H), 9.21 (s, 1H), 9.16 (d, J = 1.6 Hz, 1H), 8.15 (d, J = 5.3 Hz, 1H), 7.79 (t, J = 5.4 Hz, 1H), 7.19 (d, J = 1.7 Hz, 1H), 7.15 (s, 1H), 6.96–6.95 (m, J = 2.7 Hz, 1H), 3.40–3.37 (m, 2H), 2.85 (t, J = 6.8 Hz, 2H).

[0596] Test method: Test method B was used. Retention time = 4.11 min.

[0597] Synthesis of Example 265

[0598]

[0599] Step 1: To a suspension of (2,4-dichloro-5-(iodomethyl)pyrimidine (1 g, 3.46 mmol, 1.0 equiv) in DCM (10 mL) at -20 °C under nitrogen was added DIEPA (895 mg, 6.92 mmol, 2.0 equiv) and piperidine (295 mg, 3.46 mmol, 1.0 equiv). The reaction mixture was stirred at -20 °C for 1 h. The reaction mixture was concentrated in vacuo and purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 1) to give 2,4-dichloro-5-(piperidin-1-ylmethyl)pyrimidine (150 mg, yield 16%). LCMS (m / z): [M+H]+ calculated 246.1; found 246.1.

[0600] Step 2: To a suspension of 2,4-dichloro-5-(piperidin-1-ylmethyl)pyrimidine (100 mg, 0.41 mmol, 1.0 equiv) and NaHCO3 (102 mg, 1.22 mmol, 3.0 equiv) in DMF (3 mL) was added Int-007 (100 mg, 0.41 mmol, 1.0 equiv). The reaction mixture was stirred at 90 °C for 2 h. The mixture was filtered and the filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Inject sample in DMF, 50 min linear gradient from 10% to 90% ACN) to give Example 265 as a green solid (11.3 mg, 5.78%). LCMS (m / z): [M+H]+ calculated 457.2; found 457.2. 1 1H NMR (400 MHz, d6-DMSO) δ 11.86 (s, 1H), 8.95 (s, 1H), 8.13 (s, 0.5H), 7.71 (d, J = 7.5 Hz, 1H), 7.17 (s, 1H), 7.04 (s, 1H), 6.80 (t, J = 7.0 Hz, 1H), 3.63 (s, 2H), 3.42 (t, J = 5.6 Hz, 2H), 2.89 (t, J = 6.8 Hz, 2H), 2.48 - 2.44 (m, 4H), 1.58 - 1.52 (m, 4H), 1.45 - 1.36 (m, 2H).

[0601] Test method: Test method B was used. Retention time = 2.64 min.

[0602] Synthesis of Example 266

[0603]

[0604] Step 1: To a suspension of (2,4-dichloro-5-(iodomethyl)pyrimidine (1 g, 3.46 mmol, 1.0 equiv) in DCM (10 mL) at -20 °C under nitrogen was added DIEPA (895 mg, 6.92 mmol, 2.0 equiv) and piperidine (295 mg, 3.46 mmol, 1.0 equiv). The reaction mixture was stirred at -20 °C for 1 h. The reaction mixture was concentrated in vacuo and purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 1) to afford 2,4-dichloro-5-(piperidin-1-ylmethyl)pyrimidine (150 mg, yield 16%). LCMS (m / z): [M+H]+ calcd 246.1; found 246.1.

[0605] Step 2: To a suspension of 2,4-dichloro-5-(piperidin-1-ylmethyl)pyrimidine (100 mg, 0.41 mmol, 1.0 equiv) and NaHCO3 (102 mg, 1.22 mmol, 3.0 equiv) in DMF (3 mL) was added Int-007 (100 mg, 0.41 mmol, 1.0 equiv). The reaction mixture was stirred at 90 °C for 2 h. The mixture was filtered and the filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Samples in DMF were injected, 50 min linear gradient from 10% to 90% ACN) to give Example 266 as a white solid (26.17 mg, 13.39%). LCMS (m / z): [M+H]+ calcd 457.2; found 457.2. 1 1H NMR (400 MHz, d6-DMSO) δ 12.00 (s, 1H), 8.66 (s, 1H), 8.14 - 8.13 (m, 1H), 8.12–8.11 (m, 0.5H), 7.77 - 7.72 (m, 1H), 7.14 (s, 1H), 6.95 (s, 1H), 3.57 (s, 2H), 3.38 (td, J = 6.7, 2.1 Hz, 2H), 2.84 (t, J = 6.7 Hz, 2H), 2.40 - 2.35 (m, 4H), 1.51–1.42 (m, 4H), 1.40 - 1.30 (m, 2H).

[0606] Test method: Test method B was used. Retention time = 2.96 min.

[0607] Synthesis of Example 267

[0608]

[0609] Step 1: At 0 °C under a nitrogen atmosphere, add NaH (413 mg, 60% purity, 10.3 mmol, 1.21 equiv) to a mixture of cyclopropanol (500 mg, 8.61 mmol, 1 equiv) in DMF (5 mL) for 30 minutes. Then at 0 °C under a nitrogen atmosphere, add the mixture to a mixture of 2,4-dichloro-5-(iodomethyl)pyrimidine (1.5 g, 5.2 mmol, 1.61 equiv) in DMF (10 mL). Stir the mixture at 0 °C for 1 hour. Dilute the reaction mixture with water (20 mL) and extract with EA (30 mL × 3). Wash the organic layer with brine (15 mL) and concentrate under reduced pressure. Purify the crude product by flash column chromatography (petroleum ether:ethyl acetate = 10 / 1) to obtain 2,4-dichloro-5-(cyclopropoxymethyl)pyrimidine as a yellow solid (100 mg, yield 6.6%). LCMS (m / z): [M+H]+ calculated 219.0; found 219.0.

[0610] Step 2: Stir a mixture of 2,4-dichloro-5-(cyclopropoxymethyl)pyrimidine (70 mg, 0.35 mmol, 1 equiv), Int-007 (95 mg, 0.38 mmol), and sodium bicarbonate (80 mg, 0.96 mmol) in DMF (2 mL) at 90 °C under a nitrogen atmosphere for 1 hour. Filter the reaction mixture and purify the filtrate by preparative HPLC to obtain Example 267 (11.6 mg, yield 16%). LCMS (m / z): [M+H]+ calculated 430.1; found 430.1. 1 1H NMR (400 MHz, DMSO-d6): δ 12.06 (s, 1H), 8.75 (s, 1H), 8.18 (d, J = 5.3 Hz, 1H), 7.81 (t, J = 5.3 Hz, 1H), 7.19 (s, 1H), 6.99 (s, 1H), 4.68 (s, 2H), 3.50–3.46 (m, 1H), 3.44–3.40 (m, 2H), 2.88 (t, J = 6.8 Hz, 2H), 0.60–0.55 (m, 2H), 0.50 (dd, J = 8.5, 3.6 Hz, 2H).

[0611] Synthesis of Example 268: A mixture of 2,4-dichloro-5-(cyclopropoxymethyl)pyrimidine (70 mg, 0.35 mmol, 1 equiv), Int-007 (95 mg, 0.38 mmol), and sodium bicarbonate (80 mg, 0.96 mmol) in DMF (2 mL) was stirred at 90 °C for 1 h under a nitrogen atmosphere. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC to give Example 268 (19.63 mg, yield 25%). LCMS (m / z): [M+H]+ calculated 430.1; found 430.1. 1 1H NMR (400 MHz, DMSO-d6): δ 12.03 (s, 1H), 8.72 (s, 1H), 8.15 (d, J = 5.3 Hz, 1H), 7.76 (t, J = 5.4 Hz, 1H), 7.17 (s, 1H), 6.97 (s, 1H), 4.57 (s, 2H), 3.45–3.43 (m, 1H), 3.41 (dd, J = 5.4, 2.5 Hz, 2H), 2.88 (t, J = 6.8 Hz, 2H), 0.58–0.55 (m, 2H), 0.51–0.47 (m, 2H).

[0612] Test method: Test method B was used.

[0613] Synthesis of Example 269

[0614]

[0615] Step 1: NaH (237 mg, 60%, 5.94 mmol, 2 equiv) was added to a suspension of 2,2,2-trifluoroethanol (300 mg, 2.96 mmol, 1 equiv) in THF (6 mL) at 0 °C under nitrogen. The mixture was stirred for 0.5 h, and then the mixture was added to a solution of 2,4-dichloro-5-(iodomethyl)pyrimidine (1286 mg, 3.56 mmol, 1.2 equiv) in THF (8 mL). The mixture was stirred at 0 °C for 2 h. The reaction mixture was diluted with NH4Cl (20 mL) and extracted with EA (20 mL x 3). The organic phase was dried over anhydrous Na2SO4. After filtration, the solvent was removed in vacuo, and the residue was purified by flash column chromatography on silica gel (eluent: PE / EA = 5:1) to give 2,4-dichloro-5-((2,2,2-trifluoroethoxy)methyl)pyrimidine as a yellow oil (240 mg, yield: 61%). LCMS (m / z): [M+H]+ calculated 260.97; found 261.0.

[0616] Step 2: To a solution of Int-007 (136 mg, 0.55 mmol, 1 equiv) in DMF (1 mL) was added 2,4-dichloro-5-((2,2,2-trifluoroethoxy)methyl)pyrimidine (240 mg, 0.55 mmol, 1 equiv) and NaHCO3 (138 mg, 1.65 mmol, 3 equiv). The mixture was stirred at 50 °C under nitrogen for 2 h and then filtered. The filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMF was injected and eluted with a linear gradient from 36% to 95% ACN over 11.64 min), to give Example 269 as a white solid (19.82 mg, 7%). LCMS (m / z): [M+H]+ calculated 472.0; found 471.6. 1 1H NMR (400 MHz, d6-DMSO) δ 12.07 (s, 1H), 8.78 (s, 1H), 8.19 (d, J = 5.3 Hz, 1H), 7.81 (t, J = 5.3 Hz, 1H), 7.19 (s, 1H), 6.98 (s, 1H), 4.87 (s, 2H), 4.27 (q, J = 9.3 Hz, 2H), 3.42 (t, J = 6.8, Hz, 2H), 2.88 (t, J = 6.8 Hz, 2H).

[0617] Test method: Test method B was used. Retention time = 5.72 min.

[0618] Synthesis of Example 270: To a solution of Int-007 (136 mg, 0.55 mmol, 1 equiv) in DMF (1 mL) was added 2,4-dichloro-5-((2,2,2-trifluoroethoxy)methyl)pyrimidine (240 mg, 0.55 mmol, 1 equiv) and NaHCO3 (138 mg, 1.65 mmol, 3 equiv). The mixture was stirred at 50 °C under nitrogen for 2 h and then filtered. The filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMF was injected and eluted with a linear gradient from 36% to 95% ACN over 12.75 min), to give Example 270 as a white solid (16.22 mg, 6%). LCMS (m / z): [M+H]+ calculated 472.0; found 471.7. 1H NMR (400MHz, d6-DMSO) δ12.04(s,1H),8.74(s,1H),8.16(d,J=5.3Hz,1H),7.77(t,J=5.3Hz,1H),7.18 (s,1H),6.97(s,1H),4.75(s,2H),4.22(q,J=9.3Hz,2H),3.42(t,J=6.8Hz,2H),2.88(t,J=6.8Hz,2H).

[0619] Test Method: Used Test Method B. Retention Time = 5.82 minutes.

[0620] Synthesis of Example 271

[0621]

[0622] Step 1: To a solution of tert-butyl 8,10-dioxo-7-azaspiro[4.5]decane-7-carboxylate (200 mg, 0.75 mmol, 1 eq.) and 2-bromo-1-(2-bromo-3-fluoropyridin-4-yl)ethan-1-one (266 mg, 0.89 mmol, 1.2 eq.) in EtOH / AcOH (v / v=10 / 1, 22 mL) was added NH4OAc (575 mg, 7.45 mmol, 10 eq.) The reaction mixture was stirred at 80°C for 2 hours. The mixture was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate: petroleum ether = 1 / 1) to give tert-butyl 2'-(2-bromo-3-fluoropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (115 mg, yield 29.8%) as a yellow solid. LCMS (m / z): [M-55]+ calculated 408.0; found 408.0.

[0623] Step 2: Pd2(dba)3 (40 mg, 0.069 mmol, 0.1 eq.), t-BuXPhos (58 mg, 0.14 mmol, 0.2 eq.), KOH (77 mg, 1.38 mmol, 3 eq.) were added to a suspension of tert-butyl 2'-(2-bromo-3-fluoropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (320 mg, 0.69 mmol, 1 eq.) in dioxane / H2O (v / v=4 / 1, 20 mL). The reaction mixture was stirred at 100°C for 1 hour under microwave. The pH of the mixture was adjusted to 5-6 with 1M HCl and then extracted with EA (30 mLx3). The combined organic layers were concentrated in vacuo to give crude tert-butyl 2'-(3-fluoro-2-hydroxypyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (230 mg) as a yellow solid. LCMS (m / z): [M-55]+ Calcd. 346.1; Found 346.1.

[0624] Step 3: To a solution of tert-butyl 2'-(3-fluoro-2-hydroxypyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (170 mg, 0.42 mmol, 1 eq) and 2,4-dichloro-5-(ethoxymethyl)pyrimidine (87 mg, 0.42 mmol, 1 eq) in DMF (5 mL) was added NaHCO3 (71 mg, 0.84 mmol, 2 eq). The mixture was stirred at 90° C. for 2 h and then filtered. The filtrate was purified by flash column chromatography on silica gel (ethyl acetate: petroleum ether = 1 / 1) to give tert-butyl 2'-(2-((4-chloro-5-(ethoxymethyl)pyrimidin-2-yl)oxy)-3-fluoropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (180 mg, yield 66.9%). The product contains 4 isomers. LCMS (m / z): [M-55] + Calculated: 522.0; measured: 522.1.

[0625] Step 4: A solution of tert-butyl 2'-(2-((4-chloro-5-(ethoxymethyl)pyrimidin-2-yl)oxy)-3-fluoropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (120 mg, 0.21 mmol, 1 equiv) in DCM (10 mL) and TFA (1 mL) was stirred at 25 °C for 0.5 h. The mixture was concentrated in vacuo and the residue was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Sample injected in DMF, linear gradient from 25% to 60% ACN in 12.5 min), and then purified by chiral HPLC to afford Example 271 (7.16 mg, yield 7.21%). LCMS (m / z): [M+H]+ calculated 472.1; found 472.1. 1 1H NMR (400 MHz, DMSO-d6) δ 11.60 (s, 1H), 8.74 (s, 1H), 8.21 (d, J = 5.3 Hz, 1H), 7.95 (t, J = 5.4 Hz, 1H), 7.26 (s, 1H), 6.91 (s, 1H), 4.64 (s, 2H), 3.60 (q, J = 7.1 Hz, 2H), 3.18 (d, J = 2.1 Hz, 2H), 1.98–1.69 (m, 8H), 1.19 (t, J = 7.0, 3H).

[0626] Test method: Test method B was used.

[0627] Synthesis of Example 272

[0628]

[0629] Step 1: To a suspension of Int-0012 (10 mg, 43.62 μmol, 1.0 equiv) in DMF (0.5 mL) was added 4,6-dichloro-2-(trifluoromethyl)pyrimidine (47.33 mg, 218.12 μmol, 5.0 equiv) and K2CO3 (30.15 mg, 218.12 μmol, 5.0 equiv). The mixture was stirred at room temperature for 24 h and purified by HPLC (5-95% acetonitrile / water, containing 0.1% FA) to afford Example 272 (3.2 mg, 80% purity, 14.3% yield). LCMS (ESI): m / z = 410.0 [M+H]+. 11H NMR (400 MHz, DMSO-d6): δ 12.00 (s, 1H), 8.28 (d, J = 5.2 Hz, 1H), 7.88 (s, 1H), 7.64 (dd, J = 1.6, 5.6 Hz, 1H), 7.55 (s, 1H), 7.11 (m, 2H), 3.42 (m, 2H), 2.85 (t, J = 6.8 Hz, 2H).

[0630] Synthesis of Example 273

[0631]

[0632] Step 1: At room temperature, 2,4-dichloroquinazoline (55.4 mg, 278.6 μmol, 3.0 equiv) and K2CO3 (51.2 mg, 0.37 mmol, 4.0 equiv) were added to a suspension of Int-0012 (21.6 mg, 94.2 μmol, 1.0 equiv) in DMF / acetonitrile (2 mL, v / v = 1 / 1). The resulting mixture was stirred at 70 °C for 3 h and at room temperature for 20 h. The mixture was diluted with EtOAc and washed with water / brine. The organic layer was dried (Na2SO4) and concentrated. The residue was purified by silica gel column chromatography (0 - 10% MeOH / CH2Cl2), and then by HPLC purification (5 - 85% acetonitrile / water, containing 0.1% FA) to give Example 273 (2.7 mg, 2.5% yield). LCMS (ESI): m / z = 392.1 [M + H]+. 1 1H NMR (400 MHz, DMSO-d6): δ 12.03 (s, 1H), 8.40 (d, J = 7.6 Hz, 1H), 8.35 (d, J = 5.6 Hz, 1H), 8.13 (dt, J = 1.6, 7.2 Hz, 1H), 7.99 (d, J = 8.4 Hz, 1H), 7.84 (m, 1H), 7.69 (dd, J = 1.6, 5.2 Hz, 1H), 7.67 (s, 1H), 7.10 (m, 2H), 3.40 (m, 2H), 2.85 (t, J = 6.8 Hz, 2H).

[0633] Synthesis of Example 274

[0634]

[0635] Step 1: To a solution of 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-iodopyrimidine (148 mg, 0.36 mmol, 1.0 equiv) and 4-methyl-5-(4,4,5,5-tetramethyl-1,3-dioxolan-2-yl)oxazole (90 mg, 0.43 mmol, 1.2 equiv) in 1,4-dioxane (10 mL) and water (2 mL) was added Pd(dppf)Cl2·DCM (29 mg, 36 μmol, 0.1 equiv) and K3PO4 (229 mg, 1.08 mmol, 3.0 equiv). The reaction mixture was stirred at 55 °C overnight. The mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (0 - 33% EtOAc in PE) to give 5-(4-((4-bromopyridin-2-yl)oxy)-2-chloropyrimidin-5-yl)-4-methyloxazole (100 mg, 75%). LCMS-ESI (m / z): [M+H]+ calcd 366.95; found 367.05.

[0636] Step 2: To a solution of 5-(4-((4-bromopyridin-2-yl)oxy)-2-chloropyrimidin-5-yl)-4-methyloxazole (100 mg, 0.27 mmol, 1.0 equiv) and Int-005 (141 mg, 0.54 mmol, 2.0 equiv) in 1,4-dioxane (10 mL) and water (2 mL) was added Pd(dppf)Cl2·DCM (22 mg, 27 μmol, 0.1 equiv) and K3PO4 (172 mg, 0.81 mmol, 3.0 equiv). The reaction mixture was stirred at 65 °C for 1.5 h using a microwave reactor. The mixture was filtered and the filtrate was concentrated. The residue was purified by preparative HPLC to give Example 274 (6 mg, 5%). LCMS-ESI (m / z): [M+H]+ calcd 423.09; found 422.94. 1 1H NMR (DMSO-d6): δ 12.01 (s, 1H), 8.93 (s, 1H), 8.54 (s, 1H), 8.28 (d, J = 5.2 Hz, 1H), 7.64 - 7.63 (m, 1H), 7.57 (s, 1H), 7.13 (d, J = 2.4 Hz, 2H), 3.43 - 3.39 (m, 2H), 2.87 - 2.83 (t, J = 6.8 Hz, 2H), 2.29 (s, 3H).

[0637] Test method: Test method A was used. Retention time = 2.07 min.

[0638] Synthesis of Example 275

[0639]

[0640] Step 1: To a solution of 4-((4-bromopyridin-2-yl)oxy)-2-chloro-5-iodopyrimidine (395 mg, 0.96 mmol, 1.0 equiv) and 2-methyl-5-(4,4,5,5-tetramethyl-1,3-dioxolan-2-yl)oxazole (240 mg, 1.15 mmol, 1.2 equiv) in 1,4-dioxane (20 mL) and water (4 mL) was added Pd(dppf)Cl2·DCM (78 mg, 96 μmol, 0.1 equiv) and K3PO4 (611 mg, 2.88 mmol, 3.0 equiv). The reaction mixture was stirred at 55 °C overnight. The mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (PE solution with 0 - 50% EtOAc) to give 5-(4-((4-bromopyridin-2-yl)oxy)-2-chloropyrimidin-5-yl)-2-methyloxazole (180 mg, 51%). LCMS-ESI (m / z): [M+H]+ calculated 366.95; found 367.05.

[0641] Step 2: To a solution of 5-(4-((4-bromopyridin-2-yl)oxy)-2-chloropyrimidin-5-yl)-2-methyloxazole (100 mg, 0.27 mmol, 1.0 equiv) and Int-005 (141 mg, 0.54 mmol, 2.0 equiv) in 1,4-dioxane (10 mL) and water (2 mL) was added Pd(dppf)Cl2·DCM (22 mg, 27 μmol, 0.1 equiv) and K3PO4 (172 mg, 0.81 mmol, 3.0 equiv). The reaction mixture was stirred in a microwave reactor at 65 °C for 1.5 h. The mixture was filtered and the filtrate was concentrated. The residue was purified by preparative HPLC to give Example 275 (4 mg, 3%). LCMS-ESI (m / z): [M+H]+ calculated 423.09; found 422.96. 1 1H NMR (DMSO-d6): δ 12.03 (s, 1H), 9.02 (s, 1H), 8.31 (d, J = 5.2 Hz, 1H), 7.68 - 7.66 (m, 1H), 7.65 (s, 1H), 7.61 (d, J = 0.8 Hz, 1H), 7.13 (d, J = 2.4 Hz, 2H), 3.43 - 3.39 (m, 2H), 2.87 - 2.83 (t, J = 6.8 Hz, 2H), 2.55 (s, 3H).

[0642] Test method: Test method A used. Retention time = 2.09 minutes.

[0643] Synthesis of Example 276

[0644]

[0645] Step 1: A suspension of 2,4-dichloro-N-isopropylpyrimidine-5-carboxamide (100 mg, 0.43 mmol, 1 equiv), Int-014 (127 mg, 0.51 mmol, 1.2 equiv) and NaHCO3 (179 mg, 2.14 mmol, 5 equiv) in DMF (2 mL) was stirred at 90 °C under nitrogen for 1 h and then filtered. The filtrate was purified by preparative HPLC to give Example 276 (1.15 mg, yield 0.59%). LCMS (m / z): [M+H]+ calculated 445.11; found 445.2. 1 1H NMR (400 MHz, d6-DMSO) δ 12.02 (s, 1H), 8.84 (s, 1H), 8.50 (d, J = 7.5 Hz, 1H), 8.37 (d, J = 2.9 Hz, 1H), 7.66 (d, J = 5.5 Hz, 1H), 7.20 (s, 1H), 7.03 (s, 1H), 4.01 (dd, J = 13.8, 6.6 Hz, 1H), 3.46–3.41 (m, 2H), 2.87 (t, J = 6.8 Hz, 2H), 1.15 (d, J = 6.6 Hz, 6H).

[0646] Test method: Test method F was used.

[0647] Synthesis of Example 277

[0648]

[0649] Step 1: A suspension of 2,4-dichloro-N-isopropylpyrimidine-5-carboxamide (100 mg, 0.43 mmol, 1 equiv), Int-007 (127 mg, 0.51 mmol, 1.2 equiv) and sodium bicarbonate (179 mg, 2.14 mmol, 5 equiv) in DMF (2 mL) was stirred at 90 °C under nitrogen for 1 h and then filtered. The filtrate was purified by preparative HPLC to give Example 277 (3.64 mg, yield 1.71%). LCMS (m / z): [M+H]+ calculated 445.11; found 445.1. 11H NMR (400 MHz, d6-DMSO) δ 12.07 (s, 1H), 8.88 (s, 1H), 8.57 (d, J = 7.6 Hz, 1H), 8.14 (d, J = 5.3 Hz, 1H), 7.78 (t, J = 5.4 Hz, 1H), 7.19 (s, 1H), 6.99 (t, J = 2.5 Hz, 1H), 4.01 (dd, J = 13.8, 6.7 Hz, 1H), 3.42 (t, J = 5.8 Hz, 2H), 2.87 (t, J = 5.8 Hz, 2H), 1.18–1.09 (m, 6H).

[0650] Test method: Test method F was used.

[0651] Synthesis of Example 278

[0652]

[0653] Step 1: A solution of NaH solution (138 mg, 60% in oil, 3.46 mmol, 2 eq) was added to a solution of 2,4-dichloro-5-(iodomethyl)pyrimidine (500 mg, 1.73 mmol, 1 eq) and piperidin-2-one (206 mg, 2.08 mmol, 1.2 eq) in THF (20 mL) at 0 °C under a nitrogen atmosphere with a balloon. The reaction mixture was stirred at 0 °C for 0.5 h. Then the mixture was quenched with 1 mL of AcOH and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (ethyl acetate: petroleum ether = 1 / 2) to give the product 3 as a yellow oil (200 mg, yield 40%). LCMS (m / z): [M+H]+ calculated 260.0; found 260.0.

[0654] Step 2: NaHCO3 (194 mg, 2.3 mmol, 3 eq) was added to a solution of 1-((2,4-dichloropyrimidin-5-yl)methyl)piperidin-2-one (200 mg, 0.77 mmol, 1 eq) and Int-007 (228 mg, 0.92 mmol, 1.2 eq) in DMF (5 mL). The mixture was stirred at 90 °C for 1 h and then filtered. The filtrate was purified by preparative HPLC using a gradient of H2O (0.1% FA) / ACN (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Samples in DMF were injected, with a linear gradient from 25% to 60% ACN over 12.5 min), to give Example 278 (20.39 mg, yield 5.35%). LCMS (m / z): [M+H]+ calculated 471.1; found 471.1. 11H NMR (400 MHz, DMSO-d6) δ 12.56 (s, 1H), 8.59 (s, 1H), 8.18 (d, J = 5.3 Hz, 1H), 7.80 (t, J = 5.4 Hz, 1H), 7.19 (s, 1H), 6.99 (t, J = 2.6 Hz, 1H), 4.61 (s, 2H), 3.45–3.39 (m, 4H), 2.88 (t, J = 6.9, 2H), 2.28 (t, J = 6.3 Hz, 2H), 1.78–1.70 (m, 4H).

[0655] Test method: Test method B was used.

[0656] Synthesis of Example 279

[0657]

[0658] Step 1: To a solution of Int-007 (100 mg, 0.41 mmol) in DMF (5 mL) was added NaHCO3 (102 mg, 1.22 mmol) and 4,6-dichloro-2-(2-methoxyethoxy)pyrimidine (91 mg, 0.41 mmol). The mixture was stirred at 90 °C for 2 h under a nitrogen atmosphere. The mixture was filtered and the filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMF was injected and the linear gradient was from 35% to 65% ACN in 12.5 min), to give Example 279 (18.51 mg, purity 98.6%, yield 10.4%). LCMS (m / z): [M+H]+ calculated 434.1; found 434.2. 1 1H NMR (400 MHz, d6-DMSO) δ 12.04 (s, 1H), 8.17 (d, J = 5.3 Hz, 1H), 7.78 (t, J = 5.4 Hz, 1H), 7.27 (s, 1H), 7.18 (s, 1H), 6.99 (t, J = 2.5 Hz, 1H), 4.28 (dd, J = 5.2, 3.9 Hz, 2H), 3.54 (dd, J = 5.3, 3.7 Hz, 2H), 3.42 (td, J = 6.8, 2.2 Hz, 2H), 3.20 (s, 3H), 2.88 (t, J = 6.8 Hz, 2H).

[0659] Test method: Test method B was used.

[0660] Synthesis of Example 280

[0661]

[0662] Step 1: To a solution of tert-butyl 8,10-dioxo-7-azaspiro[4.5]decane-7-carboxylate (200 mg, 0.75 mmol, 1 eq.) and 2-bromo-1-(2-bromo-3-fluoropyridin-4-yl)ethan-1-one (266 mg, 0.89 mmol, 1.2 eq.) in EtOH / AcOH (v / v=10 / 1, 22 mL) was added NH4OAc (575 mg, 7.45 mmol, 10 eq.) The reaction mixture was stirred at 80°C for 2 hours. The mixture was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate: petroleum ether = 1 / 1) to give tert-butyl 2'-(2-bromo-3-fluoropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (115 mg, yield 29.8%). LCMS (m / z): [M-55]+ Calculated 408.0; Found 408.0.

[0663] Step 2: Pd2(dba)3 (40 mg, 0.069 mmol, 0.1 eq.), t-BuXPhos (58 mg, 0.14 mmol, 0.2 eq.), KOH (77 mg, 1.38 mmol, 3 eq.) were added to a suspension of tert-butyl 2'-(2-bromo-3-fluoropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (320 mg, 0.69 mmol, 1 eq.) in dioxane / H2O (v / v=4 / 1, 20 mL). The reaction mixture was stirred at 100°C for 1 hour under microwave. The pH of the mixture was adjusted to 5-6 with 1M HCl and then extracted with EA (30 mLx3). The combined organic layers were concentrated in vacuo to give crude tert-butyl 2'-(3-fluoro-2-hydroxypyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (230 mg). LCMS (m / z): [M-55]+ Calculated 346.1; Found 346.1.

[0664] Step 3: To a solution of tert-butyl 2'-(3-fluoro-2-hydroxypyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (170 mg, 0.42 mmol, 1 eq) and 2,4-dichloro-5-(ethoxymethyl)pyrimidine (87 mg, 0.42 mmol, 1 eq) in DMF (5 mL) was added NaHCO3 (71 mg, 0.84 mmol, 2 eq). The mixture was stirred at 90° C. for 2 h and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate: petroleum ether = 1 / 1) to give tert-butyl 2'-(2-((4-chloro-5-(ethoxymethyl)pyrimidin-2-yl)oxy)-3-fluoropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (180 mg, yield 66.9%). LCMS (m / z): [M-55]+ Calculated 522.0; Found 522.1.

[0665] Step 4: A solution of tert-butyl 2'-(2-((4-chloro-5-(ethoxymethyl)pyrimidin-2-yl)oxy)-3-fluoropyridin-4-yl)-4'-oxo-l',4'-dihydrospiro[cyclopentane-l,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (120 mg, 0.21 mmol, 1 eq) in DCM (10 mL) and TFA (1 mL) was stirred at 25 °C for 0.5 h. The mixture was concentrated in vacuo and the residue was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Sample injected in DMF, 12.5 min linear gradient from 25% to 60% ACN) and then purified by chiral HPLC to give Example 280 (7.16 mg, 7.21% yield). LCMS (m / z): [M+H]+ calcd. 472.1; found 472.1. 1 HNMR (400MHz, DMSO-d6) δ11.60(s,1H),8.74(s,1H),8.21(d,J=5.3Hz,1H),7.95(t,J=5.4Hz,1H),7.26(s,1H), 6.91(s,1H),4.64(s,2H),3.60(q,J=7.1Hz,2H),3.18(d,J=2.1Hz,2H),1.98–1.69(m,8H),1.19(t,J=7.0,3H).

[0666] Test Method: Used Test Method B. Retention Time = 1.58 minutes.

[0667] Synthesis of Example 281

[0668]

[0669] Step 1: To a solution of tert-butyl 2'-(3-fluoro-2-hydroxypyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (170 mg, 0.42 mmol, 1 equiv) and 2,4-dichloro-5-(ethoxymethyl)pyrimidine (87 mg, 0.42 mmol, 1 equiv) in DMF (5 mL) was added NaHCO3 (71 mg, 0.84 mmol, 2 equiv). The mixture was stirred at 90 °C for 2 h and then filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 1) to give tert-butyl 2'-(2-((2-chloro-5-(ethoxymethyl)pyrimidin-4-yl)oxy)-3-fluoropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (180 mg, yield 66.9%). LCMS (m / z): [M-55]+ calcd 522.0; found 522.1.

[0670] Step 2: A solution of tert-butyl 2'-(2-((4-chloro-5-(ethoxymethyl)pyrimidin-2-yl)oxy)-3-fluoropyridin-4-yl)-4'-oxo-1',4'-dihydrospiro[cyclopentane-1,7'-pyrrolo[3,2-c]pyridine]-5'(6'H)-carboxylate (120 mg, 0.21 mmol, 1 equiv) in DCM (10 mL) and TFA (1 mL) was stirred at 25 °C for 0.5 h. The mixture was concentrated in vacuo and the residue was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Sample injected in DMF, linear gradient from 25% to 60% ACN in 12.5 min), and then purified by chiral HPLC to give Example 281 (13.55 mg, yield 13.7%). LCMS (m / z): [M+H]+ calcd 472.1; found 472.1. 1HNMR (400 MHz, DMSO-d6) δ 11.59 (s, 1H), 8.70 (s, 1H), 8.18 (d, J = 5.3 Hz, 1H), 7.91 (t, J = 5.4 Hz, 1H), 7.25 (s, 1H), 6.90 (d, J = 2.9 Hz, 1H), 4.52 (s, 2H), 3.56 (q, J = 7.0 Hz, 2H), 3.17 (d, J = 2.1 Hz, 2H), 1.96–1.66 (m, 8H), 1.16 (t, J = 7.0 Hz, 3H).

[0671] Test method: Test method B was used. Retention time = 1.58 minutes.

[0672] Synthesis of Example 282

[0673]

[0674] Step 1: At -20 °C under a nitrogen atmosphere with a balloon, a solution of lithium bis(trimethylsilyl)amide (70 mL, 1 M in THF, 0.07 mol, 5 equivalents) in THF was added to a solution of tert-butyl 2,4-dioxopiperidine-1-carboxylate (3 g, 0.014 mol, 1 equivalent) and iodoethane (4.37 g, 0.028 mol, 2 equivalents) in THF (50 mL). The mixture was warmed to room temperature and stirred for 2 hours. The reaction mixture was quenched with saturated NH4Cl solution (100 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic layers were concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 1) to give tert-butyl 5-ethyl-2,4-dioxopiperidine-1-carboxylate (2.0 g, yield 52.8%). LCMS (m / z): [M - 55]+ calculated 186.1; found 186.1.

[0675] Step 2: Ammonium acetate (2.5 g, 33 mmol, 10 equivalents) was added to a solution of tert-butyl 5-ethyl-2,4-dioxopiperidine-1-carboxylate (800 mg, 3.3 mmol, 1 equivalent) and 3 (1.2 g, 3.96 mmol, 1.2 equivalents) in EtOH / AcOH (v / v = 10 / 1, 33 mL). The reaction mixture was stirred at 80 °C for 2 hours. The mixture was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 1) to give tert-butyl 2-(2-bromo-3-fluoropyridin-4-yl)-7-ethyl-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (800 mg, yield 44.1%). LCMS (m / z): [M - 55]+ calculated 382.0; found 382.0.

[0676] Step 3: To a suspension of tert-butyl 2-(2-bromo-3-fluoropyridin-4-yl)-7-ethyl-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-carboxylate (700 mg, 1.59 mmol, 1 equiv) in dioxane / H2O (v / v = 4 / 1, 20 mL) was added Pd2(dba)3 (183 mg, 0.32 mmol, 0.2 equiv), t-BuXPhos (271 mg, 0.64 mmol, 0.4 equiv), and KOH (268 mg, 4.78 mmol, 3 equiv). The reaction mixture was stirred at 100 °C for 1 h under microwave. The pH of the mixture was adjusted to 5 - 6 with 1 M HCl, and then extracted with EA (30 mL x 3). The aqueous phase was lyophilized to give the crude 7-ethyl-2-(3-fluoro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (200 mg). LCMS (m / z): [M+H]+ calculated 276.1; found 276.1.

[0677] Step 4: To a solution of 7-ethyl-2-(3-fluoro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (500 mg, 1.82 mmol, 1 equiv) and 2,4-dichloro-5-(ethoxymethyl)pyrimidine (376 mg, 1.82 mmol, 1 equiv) in DMF (5 mL) was added NaHCO3 (305 mg, 3.63 mmol, 2 equiv). The mixture was stirred at 90 °C for 2 h and then filtered. The filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMF was injected, 12.5 min linear gradient from 25% to 60% ACN), and then purified by chiral HPLC to give Example 282 (9.24 mg, yield 1.15%). LCMS (m / z): [M+H]+ calculated 446.1; found 446.1. 11H NMR (400 MHz, DMSO-d6) δ 11.93 (s, 1H), 8.75 (s, 1H), 8.20 (d, J = 5.3 Hz, 1H), 7.86 (t, J = 5.4 Hz, 1H), 7.15 (s, 1H), 6.96 (s, 1H), 4.64 (s, 2H), 3.60 (q, J = 7.0 Hz, 2H), 3.54 (dd, J = 12.0, 4.7 Hz, 1H), 3.23 (dt, J = 12.4, 3.9 Hz, 1H), 2.91–2.87 (m, 1H), 1.85–1.73 (m, 1H), 1.19 (t, J = 7.0 Hz, 3H), 0.95 (t, J = 7.4 Hz, 3H).

[0678] Test method: Test method B was used. Retention time = 1.83 minutes.

[0679] Synthesis of Example 283

[0680]

[0681] Step 1: To a solution of 7-ethyl-2-(3-fluoro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (500 mg, 1.82 mmol, 1 equiv) and 2,4-dichloro-5-(ethoxymethyl)pyrimidine (376 mg, 1.82 mmol, 1 equiv) in DMF (5 mL) was added NaHCO3 (305 mg, 3.63 mmol, 2 equiv). The mixture was stirred at 90 °C for 2 h and then filtered. The filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Samples in DMF were injected, with a linear gradient from 25% to 60% ACN over 12.5 minutes), and then purified by chiral HPLC to give Example 283 (5.89 mg, yield 0.73%). LCMS (m / z): [M+H]+ calculated 446.1; found 446.1. 11H NMR (400 MHz, DMSO-d6) δ 11.90 (s, 1H), 8.71 (s, 1H), 8.16 (d, J = 5.3 Hz, 1H), 7.82 (t, J = 5.4 Hz, 1H), 7.13 (s, 1H), 6.94 (s, 1H), 4.52 (s, 2H), 3.58–3.51 (m, 3H), 3.26–3.18 (m, 1H), 2.89 (dt, J = 12.4, 4.5 Hz, 1H), 1.84–1.71 (m, 1H), 1.65–1.53 (m, 1H), 1.16 (t, J = 7.0 Hz, 3H), 0.95 (t, J = 7.4 Hz, 3H).

[0682] Test method: Test method B was used. Retention time = 1.93 minutes.

[0683] Synthesis of Example 284

[0684]

[0685] Step 1: To a solution of 3-(2,4-dichloropyrimidin-5-yl)isoxazole (100 mg, 0.46 mmol, 1.0 equiv) in DMF (3 mL) was added Int-0014 (126 mg, 0.51 mmol, 1.1 equiv) and NaHCO3 (117 mg, 1.39 mmol, 3.0 equiv). The reaction mixture was stirred at 90 °C for 2 h. Then it was filtered. The filtrate was purified by HPLC on silica gel (acetonitrile: H2O (0.1% FA) = 30–70%) to give Example 284 (16.26 mg, yield 8%). LCMS (m / z): [M+H]+ calculated 426.8; found 427.1. 1 1H NMR (400 MHz, d6-DMSO) δ 11.97 (s, 1H), 9.19 (d, J = 1.7 Hz, 1H), 9.03 (s, 1H), 8.39 (d, J = 3.0 Hz, 1H), 7.70 (d, J = 5.6 Hz, 1H), 7.19 (s, 1H), 7.14 (d, J = 1.7 Hz, 1H), 7.03 (t, J = 2.7 Hz, 1H), 3.42 (t, J = 6.4 Hz, 2H), 2.87 (t, J = 6.8 Hz, 2H).

[0686] Test method: Test method B was used. Retention time 4.48 minutes.

[0687] Synthesis of Example 285

[0688]

[0689] Step 1: To a solution of 3-(2,4-dichloropyrimidin-5-yl)isoxazole (100 mg, 0.46 mmol, 1.0 equiv) in DMF (3 mL) was added Int-0014 (126 mg, 0.51 mmol, 1.1 equiv) and NaHCO3 (117 mg, 1.39 mmol, 3.0 equiv). The reaction mixture was stirred at 90 °C for 2 h. Then it was filtered. The filtrate was purified by HPLC on silica gel (acetonitrile: H2O (0.1% FA) = 30 - 70%) to give Example 285 (3.46 mg, yield 1.62%). LCMS (m / z): [M+H]+ calculated 427.1; found 427.1. 1 1H NMR (400 MHz, d6-DMSO) δ 11.97 (s, 1H), 9.21 (s, 1H), 9.19 (d, J = 1.7 Hz, 1H), 8.41 (d, J = 2.9 Hz, 1H), 7.73 (d, J = 5.5 Hz, 1H), 7.18 (d, J = 1.7 Hz, 2H), 7.04 (t, J = 2.5 Hz, 1H), 3.42 (td, J = 6.7, 2.2 Hz, 2H), 2.87 (t, J = 6.8 Hz, 2H).

[0690] Test method: Test method B used. Retention time = 4.48 min.

[0691] Synthesis of Example 286

[0692]

[0693] Step 1: At 50 °C under nitrogen, 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2-oxazole (41 mg, 0.21 mmol, 1 equiv), K3PO4 (134 mg, 0.63 mmol, 3 equiv) and Pd(dppf)Cl2 (15 mg, 0.021 mmol, 0.1 equiv) were added to a solution of Int-0016-A and 2-(2-((4-chloro-5-iodopyrimidin-2-yl)oxy)-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (100 mg, 0.21 mmol, 1 equiv) in dioxane / H2O = 5 / 1 (3 mL) for 2 h, and then filtered. The filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; sample injected in DMF, linear gradient from 15% to 28% ACN in 6.59 min), to obtain a mixture of Example 286 and Example 287. After lyophilization, the mixture was purified by SFC using an isocratic 40% MeOH (0.2% NH4OH) / 60% CO2 (flow rate: 12.5 / min; 5.19 min), to obtain Example 286 (6.0 mg, 7%). LCMS (m / z): [M+H]+ calculated 427.0; found 426.9. 1 1H NMR (400 MHz, d6-DMSO) δ 12.09 (s, 1H), 9.71 (s, 1H), 9.41 (s, 1H), 9.26 (s, 1H), 8.22 (d, J = 5.3 Hz, 1H), 7.84 (t, J = 5.4 Hz, 1H), 7.20 (s, 1H), 7.00 (d, J = 2.8 Hz, 1H), 3.42 (td, J = 6.8, 4.4 Hz, 2H), 2.89 (t, J = 6.8 Hz, 2H).

[0694] Test method: Test method B used. Retention time = 4.009 min.

[0695] Synthesis of Example 287

[0696]

[0697] Step 1: To a suspension of 2-(3-fluoro-2-hydroxypyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (100 mg, 0.40 mmol, 1 equiv) in DMF (2 mL) was added 2,4-dichloro-5-iodopyrimidine (122 mg, 0.45 mmol, 1.1 equiv) and NaHCO3 (102 mg, 1.21 mmol, 3 equiv). The mixture was stirred at 90 °C for 2 h. The reaction mixture was filtered and the filtrate was purified by preparative HPLC using a H2O / ACN gradient (flow rate: 40 ml / min; injection of sample in DMF) to give a mixture of 2-(2-((2-chloro-5-iodopyrimidin-4-yl)oxy)-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one and 2-(2-((4-chloro-5-iodopyrimidin-2-yl)oxy)-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (100 mg, 51%). LCMS (m / z): [M+H]+ calculated 485.96; found 486.1.

[0698] Step 2: At 50 °C under nitrogen, to a solution of 2-(2-((2-chloro-5-iodopyrimidin-4-yl)oxy)-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one and 2-(2-((4-chloro-5-iodopyrimidin-2-yl)oxy)-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (100 mg, 0.21 mmol, 1 equiv) in dioxane / H2O = 5 / 1 (3 mL) was added 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2-oxazole (41 mg, 0.21 mmol, 1 equiv), K3PO4 (134 mg, 0.63 mmol, 3 equiv) and Pd(dppf)Cl2 (15 mg, 0.021 mmol, 0.1 equiv) for 2 h, then filtered. The filtrate was purified by preparative HPLC using a H2O(0.1% FA) / ACN gradient (flow rate: 20 ml / min; sample injected in DMF, linear gradient from 15% to 28% ACN in 6.59 min), to give a mixture of Example 286 and Example 287. After lyophilization, the mixture was purified by SFC using an isocratic 40% MeOH(0.2% NH4OH) / 60% CO2 (flow rate: 12.5 / min; 5.19 min), to give Example 287 (0.63 mg, 1%). LCMS (m / z): [M+H]+ calculated 427.0; found 426.9. LCMS (m / z): [M+H]+ calculated 427.0; found 426.9. 1 H NMR (400 MHz, d6-DMSO) δ 12.06 (s, 1H), 9.52 (s, 1H), 9.15 (s, 1H), 9.00 (s, 1H), 8.17 (d, J = 5.3 Hz, 1H), 7.78 (t, J = 5.4 Hz, 1H), 7.19 (s, 1H), 6.98 (s, 1H), 3.42 (t, J = 6.8 Hz, 2H), 2.89 (t, J = 6.8 Hz, 2H).

[0699] Test method: Test method B used. Retention time = 3.922 min.

[0700] Synthesis of Example 288

[0701]

[0702] Step 1: To a solution of Int-007 (70 mg, 283 μmol, 1.0 eq) in DMF (3 mL) was added 2,4-dichloro-7,8-dihydro-5H-pyrano[4,3-d]pyrimidine (70 mg, 340 μmol, 1.2 eq) and NaHCO3 (71 mg, 849 μmol, 3.0 eq). The reaction mixture was stirred at 100 °C for 2 h. The reaction mixture was purified by preparative HPLC to give Example 288 (50 mg, 43%). LCMS-ESI (m / z): [M+H]+ calculated 416.09; found 416.02. 1 1H NMR (400 MHz, DMSO-d6) δ 12.07 (s, 1H), 8.18 (d, J = 5.3 Hz, 1H), 7.80 (t, J = 5.4 Hz, 1H), 7.21 (s, 1H), 6.99 (d, J = 2.9 Hz, 1H), 4.80 (s, 2H), 4.04 (t, J = 5.6 Hz, 2H), 3.43 (m, 2H), 2.90 (m, 4H).

[0703] Test method: Test method A used. Retention time = 2.073 min.

[0704] Synthesis of Example 289

[0705]

[0706] Step 1: To a solution of 1,3-oxazole (5 g, 72.41 mmol, 1.0 eq) in THF (60 mL) at -78 °C under nitrogen was added n-BuLi (2.5 M in THF, 29 mL, 72.5 mmol, 1.0 eq). The reaction mixture was stirred at -78 °C for 1 h. Then tris(isopropyl)(trifluoromethanesulfonyloxy)-silane (22.18 g, 72.41 mmol, 1.0 eq) was added to the reaction. The reaction mixture was warmed to room temperature and stirred for 16 h. The reaction mixture was quenched with saturated NH4Cl solution (100 mL). The mixture was extracted with ethyl acetate (200 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 1) to give 2-(triisopropylsilyl)oxazole as a yellow oil (14 g, yield 77%). LCMS (m / z): [M+H]+ calculated 226.2; found 226.2.

[0707] Step 2: At -78 °C under nitrogen, n-BuLi (2.5 M in THF, 28.3 mL, 70.75 mmol, 1.2 equiv) was added to a solution of 2-(triisopropylsilyl)oxazole (13.3 g, 59.16 mmol, 1.0 equiv) in THF (130 mL). The reaction mixture was stirred at -78 °C for 1 h. Then triisopropyl borate (12.98 g, 69.03 mmol, 1.2 equiv) was added to the reaction. The reaction mixture was stirred at -78 °C for 2 h. The reaction mixture was quenched with saturated NH4Cl solution. Extracted with ethyl acetate (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by reverse-phase column chromatography (acetonitrile: H2O = 8:1) to give product 3 (4 g, yield 23%). LCMS (m / z): [M+H]+ calculated 270.2; found 270.2.

[0708] Step 3: At room temperature under a nitrogen atmosphere, (2-(triisopropylsilyl)oxazol-5-yl)boronic acid (277 mg, 1.03 mmol, 2 equiv), K2CO3 (213 mg, 1.54 mmol, 3.0 equiv), Xantphos-Pd-G3 (49 mg, 0.051 mmol, 0.1 equiv) were added to a solution of Int-0016-A (250 mg, 0.51 mmol, 1.0 equiv) in dioxane / H2O (10 mL / 1 mL). Then the temperature was raised to 50 °C and stirred for 3 h. The mixture was filtered. The filtrate was concentrated in vacuo, and the residue was purified by reverse-phase column chromatography (acetonitrile: H2O = 2:3) to give 2-(2-((2-chloro-5-(2-(triisopropylsilyl)oxazol-5-yl)pyrimidin-4-yl)oxy)pyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (70 mg, yield 19%). LCMS (m / z): [M+H]+ calculated 565.2; found 565.2.

[0709] Step 4: To a solution of 2-(2-((2-chloro-5-(2-(triisopropylsilyl)oxazol-5-yl)pyrimidin-4-yl)oxy)pyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (70 mg, 0.12 mmol, 1.0 eq) in DMF (2 mL) was added CsF (36 mg, 0.24 mmol, 2.0 eq). The reaction mixture was stirred at 25 °C for 2 h. Then it was filtered and the filtrate was concentrated in vacuo. The residue was purified by preparative HPLC (acetonitrile:H2O (0.1% FA) = 30 - 70%) to give Example 289 (8.13 mg, yield 16%). LCMS (m / z): [M+H]+ calculated 427.1; found 427.0. 1 1H NMR (400 MHz, d6-DMSO) δ 12.09 (s, 1H), 9.14 (s, 1H), 8.73 (s, 1H), 8.22 (d, J = 5.3 Hz, 1H), 7.91 (s, 1H), 7.85 (t, J = 5.4 Hz, 1H), 7.20 (s, 1H), 7.00 (s, 1H), 3.42 (td, J = 6.8, 2.3 Hz, 2H), 2.89 (t, J = 6.8 Hz, 2H).

[0710] Test method: Test method B was used. Retention time = 4.06 minutes.

[0711] Synthesis of Example 290

[0712]

[0713] Step 1: Under a nitrogen atmosphere at room temperature, to a solution of Int-0017 (100 mg, 0.21 mmol, 1.0 eq) in dioxane / H2O (10 mL / 1 mL) was added (2-(triisopropylsilyl)oxazol-5-yl)boronic acid (111 mg, 0.41 mmol, 2.0 eq), K2CO3 (85 mg, 0.62 mmol, 3.0 eq), Xantphos-Pd-G3 (20 mg, 0.021 mmol, 0.1 eq). The mixture was heated to 50 °C and stirred for 3 h, then filtered. The filtrate was concentrated in vacuo and the residue was purified by reverse-phase column chromatography (acetonitrile:H2O = 2:3) to give 2-(2-((2-chloro-5-(2-(triisopropylsilyl)oxazol-5-yl)pyrimidin-4-yl)oxy)-5-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (40 mg, yield 33%). LCMS (m / z): [M+H]+ calculated 583.2; found 583.1.

[0714] Step 2: To a solution of 2-(2-((2-chloro-5-(2-(triisopropylsilyl)oxazol-5-yl)pyrimidin-4-yl)oxy)-5-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (40 mg, 0.069 mmol, 1.0 equiv) in DMF (2 mL) was added CsF (21 mg, 0.14 mmol, 2.0 equiv). The reaction mixture was stirred at 25 °C for 2 h and then filtered. The filtrate was concentrated in vacuo and the residue was purified by preparative HPLC (acetonitrile:H2O (0.1% FA) = 30–70%) to give Example 290 (8.17 mg, yield 27%). LCMS (m / z): [M+H]+ calculated 427.1; found 427.1. 1 1H NMR (400 MHz, d6-DMSO) δ 12.00 (s, 1H), 9.11 (s, 1H), 8.72 (s, 1H), 8.43 (d, J = 2.9 Hz, 1H), 7.84 (s, 1H), 7.75 (d, J = 5.5 Hz, 1H), 7.20 (s, 1H), 7.05 (s, 1H), 3.42 (td, J = 6.7, 2.0 Hz, 2H), 2.87 (t, J = 6.8 Hz, 2H).

[0715] Test method: Test method B was used. Retention time = 4.15 minutes.

[0716] Synthesis of Example 291

[0717]

[0718] Step 1: A suspension of 2,4-dichloro-5-(iodomethyl)pyrimidine (500 mg, 1.73 mmol, 1.0 equiv), 1H-pyrazole (106 mg, 1.55 mmol, 0.9 equiv) and K2CO3 (239 mg, 1.73 mmol) in ACN (5 mL) was stirred under nitrogen at 85 °C for 3 h. The mixture was extracted with EA (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (petroleum:ether = 2 / 1) to give 5-((1H-pyrazol-1-yl)methyl)-2,4-dichloropyrimidine (250 mg, yield 56%). LCMS (m / z): [M+H]+ calculated 229.0; found 229.0.

[0719] Step 2: A suspension of Int-007 (250 mg, 1.01 mmol, 1.0 equiv), 5-((1H-pyrazol-1-yl)methyl)-2,4-dichloropyrimidine (254 mg, 1.1 mmol, 1.1 equiv) and NaHCO3 (254 mg, 3.03 mmol, 3.0 equiv) in DMF (3 mL) was stirred at 90 °C for 2 h under nitrogen. The mixture was filtered. The filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMSO was injected, with a 15-minute linear gradient from 33% to 95% ACN), to give Example 291 (1.46 mg, yield 0.79%). LCMS (m / z): [M+H]+ calculated 440.1; found 440.1. 1 1H NMR (400 MHz, DMSO-d6) δ 12.05 (s, 1H), 8.52 (s, 1H), 8.14 (d, J = 5.4 Hz, 1H), 7.90 (d, J = 2.0 Hz, 1H), 7.76 (t, J = 5.2 Hz, 1H), 7.50 (s, 1H), 7.17 (s, 1H), 6.97 (s, 1H), 6.31–6.28 (m, 1H), 5.45 (s, 2H), 3.41 (t, J = 7.5 Hz, 2H), 2.87 (t, J = 6.6 Hz, 2H).

[0720] Test method: Test method B was used.

[0721] Synthesis of Example 292

[0722]

[0723] Step 1: To a solution of 4-(tributylstannyl)-2-(triisopropylsilyl)-1,3-oxazole (794 mg, 1.53 mmol, 3 equiv) in anhydrous ACN (10 mL) was added Int-0016-A (250 mg, 0.51 mmol, 1 equiv) and bis(triphenylphosphine)palladium(II) chloride (37 mg, 0.051 mmol, 0.1 equiv). The mixture was stirred at 85 °C for 18 h. Then the mixture was concentrated and the residue was purified by flash chromatography (C-18 reverse phase column, water / ACN, 1:1) to give 2-(2-((2-chloro-5-(2-(triisopropylsilyl)oxazol-4-yl)pyrimidin-4-yl)oxy)-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one as a pale yellow solid (220 mg, purity 90%, yield 73%). LCMS (m / z): [M+H]+ calculated 583.2; found 583.2.

[0724] Procedure: To a solution of 2-[2-({2-chloro-5-[2-(triisopropylsilyl)-1,3-oxazol-4-yl]pyrimidin-4-yl}oxy)-3-fluoropyridin-4-yl]-1H,5H,6H,7H-pyrrolo[3,2-c]pyridin-4-one (220 mg, 0.38 mmol, 1 equiv) in DMF (10 mL) was added CsF (143 mg, 0.95 mmol, 2.5 equiv). The mixture was stirred at 25 °C for 2 h under a nitrogen atmosphere. The mixture was filtered and the filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMF was injected and the linear gradient was from 25% to 60% ACN in 12.5 min), to give Example 292 (39 mg, purity 97.26%, yield 23%). LCMS (m / z): [M+H]+ calculated 427.1; found 427.1. 1 H NMR (400 MHz, d6-DMSO) δ 12.07 (s, 1H), 9.22 (s, 1H), 8.84 (s, 1H), 8.70 (d, J = 0.7 Hz, 1H), 8.22 (d, J = 5.3 Hz, 1H), 7.84 (t, J = 5.4 Hz, 1H), 7.18 (s, 1H), 7.00 (t, J = 2.5 Hz, 1H), 3.42 (td, J = 6.8, 2.3 Hz, 2H), 2.89 (t, J = 6.8 Hz, 2H).

[0725] Test method: Test method B was used. Retention time = 4.610 min.

[0726] Synthesis of Example 293

[0727]

[0728] Step 1: At -78 °C, n-BuLi (2.5 M in THF, 43.8 mL, 109.5 mmol, 1.2 equiv) was added to a solution of 5-bromo-2,4-dimethoxypyrimidine (20 g, 91.32 mmol, 1.0 equiv) in THF (200 mL). The reaction mixture was stirred at -78 °C for 1 h, then N,N-dimethylformamide (8.01 g, 109.58 mmol, 1.2 equiv) was added to the reaction mixture, and the reaction mixture was stirred at -78 °C for another 1 h. The reaction mixture was quenched with saturated NH4Cl solution (100 mL). The mixture was extracted with ethyl acetate (300 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 4) to give 2,4-dimethoxypyrimidine-5-carbaldehyde (6.65 g, yield 39%). LCMS (m / z): [M+H]+ calculated 169.05; found 169.1.

[0729] Step 2: Hydroxylamine hydrochloride (4.12 g, 59.26 mmol, 1.5 equiv) and TEA (6 g, 59.26 mmol, 1.5 equiv) were added to a solution of 2,4-dimethoxypyrimidine-5-carbaldehyde (6.65 g, 39.51 mmol, 1.0 equiv) in EtOH (70 mL). The reaction mixture was stirred at 25 °C for 3 h and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 4) to give (E)-2,4-dimethoxypyrimidine-5-carbaldehyde oxime (4.9 g, yield 61%). LCMS (m / z): [M+H]+ calculated 184.06; found 184.1.

[0730] Step 3: At 0 °C, NCS (4.38 g, 32.82 mmol, 1.1 equiv) was added to a solution of (E)-2,4-dimethoxypyrimidine-5-carbaldehyde oxime (3 g, 16.41 mmol, 1.0 equiv) in DMF (30 mL). The reaction mixture was stirred at 0 °C for 2 h. Then the mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the crude product was used in the next step without any further purification. The crude product (3.2 g, yield 70%) was obtained. LCMS (m / z): [M-35]+ calculated 218.03; found 218.1.

[0731] Step 4: To a solution of (Z)-N-hydroxy-2,4-dimethoxypyrimidine-5-carboximidoyl chloride (3.2 g, 14.69 mmol, 1.0 equiv) in DMF (30 mL) was added (1E)-1-ethoxyprop-1-ene (2.53 g, 29.38 mmol, 2.0 equiv) and NaHCO3 (2.47 g, 29.38 mmol, 2.0 equiv). The reaction mixture was stirred at 25 °C for 16 h. The reaction mixture was quenched with H2O. Extraction was performed with ethyl acetate (100 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 3) to give 3-(2,4-dimethoxypyrimidin-5-yl)-5-ethoxy-4-methyl-4,5-dihydroisoxazole (2.2 g, yield 51%). LCMS (m / z): [M+H]+ calculated 268.3; found 268.1.

[0732] Step 5: A solution of 3-(2,4-dimethoxypyrimidin-5-yl)-5-ethoxy-4-methyl-4,5-dihydroisoxazole (2.2 g, 8.20 mmol) in TFA (20 mL) was stirred at 80 °C for 3 h. The reaction solution was concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 1 / 3) to give 3-(2,4-dimethoxypyrimidin-5-yl)-4-methylisoxazole (1.74 g, yield 86%). LCMS (m / z): [M+H]+ calculated 222.1; found 222.1.

[0733] Step 6: To a solution of 3-(2,4-dimethoxypyrimidin-5-yl)-4-methylisoxazole (1.74 g, 7.86 mmol, 1.0 equiv) in DCM (20 mL) at 0 °C was added BBr3 (7.92 g, 31.44 mmol, 4.0 equiv). The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was quenched with NaHCO3 solution, and extraction was performed with ethyl acetate (50 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by silica gel reverse phase column (acetonitrile:water = 1 / 9) to give 5-(4-methylisoxazol-3-yl)pyrimidine-2,4-diol (500 mg, yield 29%). LCMS (m / z): [M+H]+ calculated 194.05; found 194.0.

[0734] Step 7: A solution of 5-(4-methylisoxazol-3-yl)pyrimidine-2,4-diol (200 mg, 1.04 mmol, 1.0 equiv) in POCl3 / DIEA = 10:1 (11 mL) was stirred at 120 °C for 2 h. The reaction solution was concentrated in vacuo. The residue was diluted with DCM (20 mL) and saturated aqueous sodium bicarbonate solution (20 mL). After separation, the organic phase was dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography on silica gel (ethyl acetate:petroleum ether = 2 / 3) to give 3-(2,4-dichloropyrimidin-5-yl)-4-methylisoxazole (150 mg, yield 57%). LCMS (m / z): [M+H]+ calculated 229.98; found 229.9.

[0735] Step 8: To a solution of 3-(2,4-dichloropyrimidin-5-yl)-4-methylisoxazole (150 mg, 0.65 mmol, 1.0 equiv) in DMF (5 mL) was added Int-007 (193 mg, 0.78 mmol, 1.2 equiv) and NaHCO3 (164 mg, 1.96 mmol, 3.0 equiv). The reaction mixture was stirred at 25 °C for 16 h. Then it was filtered. The filtrate was concentrated in vacuo, and the residue was purified by HPLC on silica gel (acetonitrile:H2O (0.1% FA) = 30 - 70%) to give Example 293 (12.24 mg, yield 4%). LCMS (m / z): [M+H] + Calculated 441.08; found 441.2. 1 1H NMR (400 MHz, d6-DMSO) δ 12.06 (s, 1H), 8.95 (s, 2H), 8.19 (d, J = 5.3 Hz, 1H), 7.82 (t, J = 5.4 Hz, 1H), 7.19 (s, 1H), 7.00 (d, J = 2.4 Hz, 1H), 3.42 (td, J = 6.8, 2.3 Hz, 2H), 2.89 (t, J = 6.8 Hz, 2H), 2.12 (d, J = 0.9 Hz, 3H).

[0736] Testing method: Testing method B was used. Retention time = 4.83 min.

[0737] Synthesis of Example 294

[0738]

[0739] Step 1: At room temperature, under a nitrogen atmosphere with a balloon, 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (39 mg, 0.20 mmol, 1.2 eq), K2CO3 (68 mg, 0.49 mmol, 3.0 eq), Pd(PPh3)4 (19 mg, 0.016 mmol, 0.1 eq) were added to a solution of Int-0017 (80 mg, 0.16 mmol, 1.0 eq) in dioxane / H2O (10 mL / 1 mL). Then the temperature was raised to 50 °C and stirred for 4 hours, and then filtered. The filtrate was concentrated in vacuo, and the residue was purified by HPLC on silica gel (acetonitrile:H2O (0.1% FA) = 30 - 70%) to give Product Example 294 (9.35 mg, yield 13%). LCMS (m / z): [M+H]+ calculated 427.06; found 427.0. 1 1H NMR (400 MHz, d6-DMSO) δ 11.96 (s, 1H), 9.60 (s, 1H), 9.34 (s, 1H), 9.17 (s, 1H), 8.39 (d, J = 2.9 Hz, 1H), 7.70 (d, J = 5.5 Hz, 1H), 7.16 (s, 1H), 7.00 (d, J = 2.4 Hz, 1H), 3.38 (td, J = 6.5, 1.8 Hz, 2H), 2.82 (t, J = 6.8 Hz, 2H).

[0740] Test method: Test method B was used. Retention time = 4.27 minutes.

[0741] Synthesis of Example 295

[0742]

[0743] Step 1: A suspension of EtOH (3.45 g, 0.075 mol, 10.0 eq) and NaH (3 g, 60%, 0.075 mol, 10.0 eq) was stirred at 0 °C for 0.5 h under nitrogen. Then a solution of 6-(chloromethyl)pyrimidine-2,4-diol (1.2 g, 7.5 mmol, 1.0 eq) in THF (8 mL) was added. The reaction mixture was stirred at 45 °C for 16 h. The mixture was adjusted to pH = 7 with saturated HCl solution, forming a yellow precipitate, which was collected by filtration. The filter cake was purified by C18 (acetonitrile:H2O = 30 - 70%) to give 6-(ethoxymethyl)pyrimidine-2,4-diol (0.6 g, yield 42%). LCMS (m / z): [M+H]+ calculated 171.1; found 171.1.

[0744] Step 2: Dissolve 6-(ethoxymethyl)pyrimidine-2,4-diol (600 mg, 3.52 mmol) in POCl3 / DIEA (v / v = 10 / 1, 6 mL), and stir at 120 °C for 2 h. Then concentrate the mixture, dilute the residue with DCM (20 mL) and wash with saturated NaHCO3 solution (20 mL). Concentrate the organic layer, and purify the residue by flash column chromatography on silica gel (petroleum: ether = 2 / 1) to obtain 2,4-dichloro-6-(ethoxymethyl)pyrimidine (600 mg, yield 74%). LCMS (m / z): [M+H]+ calculated 207.0; found 207.1.

[0745] Step 3: Suspend Int-007 (200 mg, 0.81 mmol, 1.0 equiv), 2,4-dichloro-6-(ethoxymethyl)pyrimidine (184 mg, 0.89 mmol, 1.1 equiv) and NaHCO3 (203 mg, 2.4 mmol, 3.0 equiv) in DMF (4 mL) and stir at 90 °C for 2 h under nitrogen. Then filter it, and purify the filtrate by preparative HPLC using H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. Inject the sample in DMSO, 15 min linear gradient from 33% to 95% ACN) to obtain Example 295 (24.86 mg, yield 7%). LCMS (m / z): [M+H]+ calculated 418.1; found 418.2. 1 1H NMR (400 MHz, DMSO) δ 12.06 (s, 1H), 8.17 (d, J = 5.3 Hz, 1H), 7.79 (t, J = 5.4 Hz, 1H), 7.30 (s, 1H), 7.19 (s, 1H), 6.99 (s, 1H), 4.58 (s, 2H), 3.65–3.57 (m, 2H), 3.42 (t, J = 6.8, 4.5 Hz, 2H), 2.88 (t, J = 6.7 Hz, 2H), 1.20 (t, J = 7.0 Hz, 3H).

[0746] Test method: Test method B used. Retention time = 4.768 min.

[0747] Synthesis of Example 296

[0748]

[0749] Step 1: A suspension of Int-007 (200 mg, 0.81 mmol, 1.0 eq), 2,4-dichloro-6-(ethoxymethyl)pyrimidine (184 mg, 0.89 mmol, 1.1 eq) and NaHCO3 (203 mg, 2.4 mmol, 3.0 eq) in DMF (4 mL) was stirred under nitrogen at 90 °C for 2 h. Then it was filtered and the filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample was injected in DMSO, 15 min linear gradient from 33% to 95% ACN), to give Example 296 (8.62 mg, yield 2.3%). LCMS (m / z): [M+H]+ calculated 418.1; found 418.0. 1 1H NMR (400 MHz, DMSO) δ 12.03 (s, 1H), 8.15 (d, J = 5.3 Hz, 1H), 7.76 (t, J = 5.3 Hz, 1H), 7.44 (s, 1H), 7.18 (brs, 1H), 6.97 (d, J = 2.6 Hz, 1H), 4.50 (s, 2H), 3.60–3.50 (m, 2H), 3.42 (t, J = 6.7, 2.3 Hz, 2H), 2.88 (t, J = 6.8 Hz, 2H), 1.15 (t, J = 7.0 Hz, 3H).

[0750] Test method: Test method B was used. Retention time = 4.768 min.

[0751] Synthesis of Example 297

[0752]

[0753] Step 1: To a suspension of Int-0016-A (60 mg, 0.12 mmol, 1 eq) and 2 (31 mg, 0.15 mmol, 1.5 eq) in dioxane / H2O (v / v = 4 / 1, 5 mL) was added Pd(dppf)Cl2 (9 mg, 0.012 mmol, 0.1 eq) and K2CO3 (34 mg, 0.25 mmol, 2 eq). The mixture was stirred at 60 °C for 2 h and then filtered. The filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample was injected in DMF, 12.5 min linear gradient from 25% to 60% ACN), to give Example 297 (12.85 mg, yield 23.16%). LCMS (m / z): [M+H]+ calculated 440.1; found 440.1. 11H NMR (400 MHz, DMSO-d6) δ 12.06 (s, 1H), 8.87 (s, 1H), 8.17 (d, J = 5.3 Hz, 1H), 7.80 (t, J = 5.4 Hz, 1H), 7.59 (d, J = 1.9 Hz, 1H), 7.18 (s, 1H), 6.99 (s, 1H), 6.61 (d, J = 1.9 Hz, 1H), 3.9 (s, 3H), 3.44–3.40 (m, 2H), 2.88 (t, J = 6.8, 2H).

[0754] Test method: Test method B was used.

[0755] Synthesis of Example 298

[0756]

[0757] Step 1: A mixture of Int-0012 (16.3 mg, 71.11 μmol, 1.0 equiv), 2,4-dichloro-6-fluoroquinazoline (23.15 mg, 106.66 μmol, 1.5 equiv) and K2CO3 (39.31 mg, 284.43 μmol, 4.0 equiv) in a mixture of acetonitrile (1 mL) and DMF (1 mL) was stirred at 60 °C for 2 h and at room temperature for 16 h. The mixture was diluted with EtOAc and washed with water, dried (Na2SO4), and filtered. The residue was purified by HPLC (5 - 95% acetonitrile / water, containing 0.1% FA) to give Example 298 (3.4 mg, 11% yield). LCMS (ESI): m / z = 410.1 [M+H]+. 1 1H-NMR (400 mHz, DMSO-d6) δ 12.03 (s, 1H), 8.35 (d, J = 5.2 Hz, 1H), 8.17 (dd, J = 2.4, 8.0 Hz, 1H), 8.09 (m, 2H), 8.70 (d, J = 1.6 Hz, 1H), 8.68 (s, 1H), 7.10 (m, 2H), 6.61 (brs, 1H), 3.30 (m, 2H), 2.84 (t, J = 6.8 Hz, 2H).

[0758] Synthesis of Example 299

[0759]

[0760] Step 1: A suspension of 2,4-dichloro-5-(iodomethyl)pyrimidine (500 mg, 1.73 mmol, 1.0 equiv), 1H-pyrazole (106 mg, 1.55 mmol, 0.9 equiv) and K2CO3 (239 mg, 1.73 mmol) in ACN (5 mL) was stirred at 85 °C under nitrogen for 3 h. The mixture was extracted with EA (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (petroleum: ether = 2 / 1) to give 5-((1H-pyrazol-1-yl)methyl)-2,4-dichloropyrimidine (250 mg, yield 56%). LCMS (m / z): [M+H]+ calculated 229.0; found 229.0.

[0761] Step 2: A suspension of Int-007 (250 mg, 1.01 mmol, 1.0 equiv), 5-((1H-pyrazol-1-yl)methyl)-2,4-dichloropyrimidine (254 mg, 1.1 mmol, 1.1 equiv) and NaHCO3 (254 mg, 3.03 mmol, 3.0 equiv) in DMF (3 mL) was stirred at 90 °C under nitrogen for 2 h. The mixture was filtered. The filtrate was purified by preparative HPLC using a H2O (0.1% FA) / ACN gradient (flow rate: 20 ml / min; wavelength: 214 nm / 254 nm. The sample in DMSO was injected, 15 min linear gradient from 33% to 95% ACN) to give Example 299 (6.69 mg, yield 3.6%). LCMS (m / z): [M+H]+ calculated 440.1; found 440.0. 1 1H NMR (400 MHz, DMSO-d6) δ 12.04 (s, 1H), 8.55 (s, 1H), 8.17 (d, J = 5.3 Hz, 1H), 7.92 (d, J = 2.2 Hz, 1H), 7.80 (t, J = 5.4 Hz, 1H), 7.52 (d, J = 1.6 Hz, 1H), 7.19 (brs, 1H), 6.96 (d, J = 2.9 Hz, 1H), 6.31 (t, J = 2.0 Hz, 1H), 5.54 (s, 2H), 3.41 (t, J = 6.8, 2H), 2.88 (t, J = 6.8 Hz, 2H).

[0762] Test method: Test method B was used.

[0763] Synthesis of Example 300

[0764]

[0765] Step 1: To a solution of (1-(2-methoxyethyl)-1H-pyrazol-4-yl)boronic acid SM1 (251 mg, 1.48 mmol, 1.1 eq) and 5-bromo-2-chloro-4-methoxypyrimidine SM2 (300 mg, 1.34 mmol, 1.0 eq) in 1,4-dioxane (10 mL) and water (1 mL) was added Pd(dppf)Cl2·DCM (100 mg, 0.13 mmol, 0.1 eq) and Cs2CO3 (875 mg, 2.68 mmol, 2.0 eq). The mixture was stirred overnight at 95 °C under nitrogen and purified by flash column chromatography, eluting with DCM and MeOH (10:1) to give 2-chloro-4-methoxy-5-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)pyrimidine (254 mg, 0.94 mmol, 70%). LCMS-ESI (m / z): [M+H]+ calculated 269.07; found 269.05.

[0766] Step 2: To a solution of 2-chloro-4-methoxy-5-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)pyrimidine (254 mg, 0.94 mmol, 1.0 eq) in AcOH (5.0 mL) was added NaI (425 mg, 2.84 mmol, 3.0 eq). The reaction mixture was stirred at 110 °C for 2 h and the mixture was concentrated. DCM (10 mL) was added, the mixture was stirred for 1 h and filtered to give 5-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)pyrimidine-2,4-diol (340 mg, 1.44 mmol, crude). LCMS-ESI (m / z): [M+H]+ calculated 237.09; found 237.01.

[0767] Step 3: At 0 °C, to a solution of 5-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)pyrimidine-2,4-diol (340 mg, 1.44 mmol, 1.0 eq) in POCl3 (5.0 mL) was added DIEA (558 mg, 4.32 mmol, 3.0 eq). The reaction mixture was stirred at 100 °C for 1 h. The mixture was concentrated, the pH was adjusted to 7 - 8 with aqueous NaHCO3, extracted with EA (10 mL × 3) and purified by flash column chromatography to give 2,4-dichloro-5-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)pyrimidine (40 mg, 0.15 mmol, 10%). LCMS-ESI (m / z): [M+H]+ calculated 273.02; found 272.96.

[0768] Step 4: To a solution of Int-007 (18 mg, 0.07 mmol, 1.0 eq) in DMF (2 mL) was added NaHCO3 (12 mg, 0.15 mmol, 2.0 eq). The reaction mixture was stirred at 100 °C for 0.5 h, then 2,4-dichloro-5-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)pyrimidine (20 mg, 0.07 mmol, 1.0 eq) was added. The mixture was stirred at 100 °C for 1.5 h. The mixture was purified by preparative HPLC to give Example 300 (1.6 mg, 3.3 μmol, 5%). LCMS-ESI (m / z): [M-H]- calculated 482.12; found 482.39. 1 1H NMR (DMSO-d6): δ 12.09 (s, 1H), 9.16 (s, 1H), 8.45 (s, 1H), 8.22 - 8.20 (m, 2H), 7.84 - 7.82 (m, 1H), 7.20 (s, 1H), 7.00 (s, 1H), 4.32 (t, J = 7.1 Hz, 2H), 3.72 (t, J = 7.1 Hz, 2H), 3.44 - 3.43 (m, 2H), 3.23 (s, 3H), 2.91 - 2.88 (m, 2H).

[0769] Test method: Test method B was used. Retention time = 2.13 min.

[0770] Synthesis of Example 301

[0771]

[0772] Step 1: To a round-bottom flask were added 2,4-dimethoxy-5-(1-methyl-1H-pyrazol-4-yl)pyrimidine (500 mg, 2.28 mmol, 1.0 eq), (1-methylpyrazol-4-yl)boronic acid (575 mg, 4.56 mmol, 2.0 eq), Pd(dppf)Cl2·CH2Cl2 (185 mg, 0.22 mmol, 0.1 eq) and K2CO3 (946 mg, 6.84 mmol, 3 eq). Then 1,4-dioxane (10 mL) and water (1 mL) were added. The reaction was stirred at 90 °C for 16 h under a nitrogen atmosphere. The reaction mixture was filtered, the filtrate was concentrated and purified by silica gel column (0 - 4% MeOH in DCM) to give 2,4-dimethoxy-5-(1-methyl-1H-pyrazol-4-yl)pyrimidine 1 (500 mg, 99%). LCMS-ESI (m / z): [M+H]+ calculated 221.10; found 221.07.

[0773] Step 2: To a solution of 2,4-dimethoxy-5-(1-methyl-1H-pyrazol-4-yl)pyrimidine (500 mg, 2.27 mmol, 1.0 equiv) in AcOH (10 mL) was added NaI (1.02 g, 6.81 mmol, 3 equiv), and the mixture was refluxed at 110 °C for 2 h. The solvent was evaporated, and the residue was diluted with saturated Na2S2O3 solution (20 mL). The solid was filtered and washed with water (10 mL) to give 5-(1-methyl-1H-pyrazol-4-yl)pyrimidine-2,4-diol (410 mg, 93%). LCMS-ESI (m / z): [M+H]+ calculated 193.06; found 193.20.

[0774] Step 3: To a solution of 5-(1-methyl-1H-pyrazol-4-yl)pyrimidine-2,4-diol (390 mg, 2.03 mmol, 1.0 equiv) in POCl3 (5 mL) was added DIEA (1.31 g, 10.15 mmol, 5.0 equiv), and the mixture was refluxed at 110 °C for 2 h. Most of the solvent was evaporated, and the residue was added dropwise to saturated NaHCO3 solution (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (100 mL) and dried over Na2SO4. The mixture was concentrated and purified by silica gel column (0 - 30% EtOAc in petroleum ether) to give 2,4-dichloro-5-(1-methyl-1H-pyrazol-4-yl)pyrimidine (320 mg, 68%). LCMS-ESI (m / z): [M+H]+ calculated 229.00; found 228.98.

[0775] Step 4: To a solution of Int-003 (100 mg, 0.4 mmol, 1.0 equiv) in DMF (0.5 mL) was added NaHCO3 (68 mg, 0.8 mmol, 2.0 equiv), and the mixture was stirred at 100 °C for 30 min. Then a solution of 2,4-dichloro-5-(1-methyl-1H-pyrazol-4-yl)pyrimidine (93 mg, 0.4 mmol, 1.0 equiv) in DMF (0.5 mL) was added to the reaction mixture. The reaction mixture was then stirred at 100 °C for another 2 h. The reaction mixture was purified by preparative HPLC to give Example 301 as a yellow solid (50 mg, 29%). LCMS-ESI (m / z): [M+H]+ calculated 440.10; found 439.99. 1HNMR (400 MHz, DMSO-d6) δ 12.06 (s, 0.86H), 12.03 (s, 0.14H), 9.15 (s, 0.86H), 8.93 (s, 0.14H), 8.44 (s, 0.86H), 8.29 (s, 0.14H), 8.21 (d, J = 5.2 Hz, 0.86H), 8.17 (d, J = 0.8 Hz, 0.86H), 8.16 (d, J = 5.2 Hz, 0.14H), 7.95 (d, J = 0.8 Hz, 0.14H), 7.82 (t, J = 5.2 Hz, 0.86H), 7.77 (t, J = 5.2 Hz, 0.14H), 7.21 - 7.19 (m, 1H), 7.01 - 6.98 (m, 1H), 3.92 (s, 3H), 3.45 - 3.41 (dt, J = 6.8, 2.4 Hz, 2H), 2.89 (t, J = 6.8 Hz, 2H).

[0776] Test method: Test method B was used, retention time = 2.08 minutes.

[0777] Synthesis of Example 302

[0778]

[0779] Step 1: To a solution of 5-bromo-2-chloro-4-methoxypyrimidine (150 mg, 0.67 mmol, 1.0 equiv) and (1,3-dimethyl-1H-pyrazol-4-yl)boronic acid (113 mg, 0.80 mmol, 1.2 equiv) in 1,4-dioxane (10 mL) and water (1 mL) was added Pd(dppf)Cl2·DCM (27 mg, 0.03 mmol, 0.05 equiv) and Cs2CO3 (256 mg, 1.34 mmol, 2.0 equiv). The mixture was stirred overnight at 90 °C in nitrogen and purified by flash column chromatography, eluting with dichloromethane and methanol (10:1) to give 2-chloro-5-(1,3-dimethyl-1H-pyrazol-4-yl)-4-methoxypyrimidine (2) (113 mg, 73%). LCMS-ESI (m / z): [M+H]+ calculated 239.07; found 239.03.

[0780] Step 2: To a solution of 2-chloro-5-(1,3-dimethyl-1H-pyrazol-4-yl)-4-methoxypyrimidine (117 mg, 0.49 mmol, 1.0 equiv) in AcOH (3 mL) was added NaI (220 mg, 1.47 mmol, 3.0 equiv), and the mixture was stirred at 110 °C for 1 h. The mixture was concentrated and dichloromethane (6 mL) was added. The mixture was stirred for 1 h and filtered to give 5-(1,3-dimethyl-1H-pyrazol-4-yl)pyrimidine-2,4-diol (80 mg, 79%). LCMS-ESI (m / z): [M+H]+ 207.09; found 207.06.

[0781] Step 3: To a solution of 5-(1,3-dimethyl-1H-pyrazol-4-yl)pyrimidine-2,4-diol (520 mg, 2.50 mmol, 1.0 equiv) in POCl3 (5 mL) at 0 °C was added DIEA (1.6 g, 12.60 mmol, 5.0 equiv), and the mixture was stirred at 110 °C for 1 h. The mixture was concentrated, and the pH was adjusted to 7 - 8 with aqueous NaHCO3. The mixture was extracted with EA (15 mL × 3) and purified by flash column chromatography, eluting with dichloromethane and methanol (10:1) to give 2,4-dichloro-5-(1,3-dimethyl-1H-pyrazol-4-yl)pyrimidine (111 mg, 18%). LCMS-ESI (m / z): [M+H]+ calculated 243.01; found 242.98.

[0782] Step 4: To a solution of Int-003 (20 mg, 0.08 mmol, 1.0 equiv) in DMF (1.5 mL) was added NaHCO3 (14 mg, 0.16 mmol, 2.0 equiv), and the mixture was stirred at 100 °C for 0.5 h. 2,4-Dichloro-5-(1,3-dimethyl-1H-pyrazol-4-yl)pyrimidine (20 mg, 0.08 mmol, 1.0 equiv) was added and the mixture was stirred at 100 °C for 2 h. The mixture was purified by preparative HPLC to give Example 302 (2.0 mg, 5%). LCMS-ESI (m / z): [M+H]+ 454.12; found 454.30. 1 1H NMR (DMSO-d6): δ 12.07 (s, 1H), 8.78 (s, 1H), 8.67 (s, 1H), 8.19 (d, J = 5.6 Hz, 1H), 8.08 (s, 1H), 7.81 (t, J = 5.2 Hz, 1H), 7.19 (s, 1H), 7.00 (s, 1H), 3.84 (s, 3H), 3.45 - 3.41 (m, 2H), 2.89 (t, J = 7.2 Hz, 2H), 2.32 (s, 3H).

[0783] Test method: Test method B was used. Retention time = 2.12 minutes.

[0784] Synthesis of Example 303

[0785]

[0786] Step 1: Trimethylsilylacetylene (59 mg, 0.60 mmol, 5 eq), Pd(PPh3)4 (55 mg, 0.05 mmol, 0.4 eq), CuI (18 mg, 0.09 mmol, 0.8 eq), and TEA (48 mg, 0.48 mmol, 4.0 eq) were added to a solution of Example 222 (50 mg, 0.12 mmol, 1.0 eq) in dichloromethane (5 mL). The mixture was stirred at 50 °C under nitrogen for 24 h. The reaction mixture was concentrated in vacuo and purified by silica gel column chromatography, eluting with dichloromethane and methanol (10:1) to give 2-(2-((5-(ethoxymethyl)-2-((trimethylsilyl)ethynyl)pyrimidin-4-yl)oxy)-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (1) (61 mg, 99%). LCMS-ESI (m / z): [M+H]+ calculated 480.19; found 480.03.

[0787] Step 2: K2CO3 (78 mg, 0.57 mmol, 2.0 eq) was added to a solution of 2-(2-((5-(ethoxymethyl)-2-((trimethylsilyl)ethynyl)pyrimidin-4-yl)oxy)-3-fluoropyridin-4-yl)-1,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one (136 mg, 0.28 μmol, 1.0 eq) in MeOH (2.5 mL). The mixture was stirred at room temperature for 0.5 h. The mixture was purified by preparative HPLC to give Example 303 (42 mg, 35%). LCMS-ESI (m / z): [M+H]+ 408.15; found 408.17. 1 1H NMR (400 MHz, DMSO-d6) δ 12.05 (s, 1H), 8.78 (s, 1H), 8.17 (d, J = 5.2 Hz, 1H), 7.79 (t, J = 5.6 Hz, 1H), 7.19 (s, 1H), 6.99 (d, J = 2.8 Hz, 1H), 4.67 (s, 2H), 4.39 (s, 1H), 3.61 (q, J = 7.2 Hz, 2H), 3.45 - 3.41 (m, 2H), 2.89 (t, J = 6.8 Hz, 2H), 1.19 (t, J = 7.2 Hz, 3H).

[0788] Test method: Test method B was used. Retention time = 2.17 minutes.

[0789] Synthesis of Example 304

[0790]

[0791] Step 1: To a solution of 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (262 mg, 1.07 mmol, 1.2 equiv) and 5-bromo-2-chloro-4-methoxypyrimidine (200 mg, 0.90 mmol, 1.0 equiv) in 1,4-dioxane (5.0 mL) and water (0.5 mL) was added Pd(dppf)Cl2·DCM (73 mg, 0.09 mmol, 0.1 equiv) and Cs2CO3 (583 mg, 1.79 mmol, 2.0 equiv). The mixture was stirred overnight at 95 °C under nitrogen and purified by flash column chromatography, eluting with petroleum ether and ethyl acetate (3:1) to give 2-chloro-5-(1-(difluoromethyl)-1H-pyrazol-4-yl)-4-methoxypyrimidine as a white solid (50 mg, 0.19 mmol, 21%). LCMS-ESI (m / z): [M+H]+ calculated 261.03; found 260.98.

[0792] Step 2: To a solution of 2-chloro-5-(1-(difluoromethyl)-1H-pyrazol-4-yl)-4-methoxypyrimidine (50 mg, 0.19 mmol, 1.0 equiv) in AcOH (1.0 mL) was added NaI (86 mg, 0.57 mmol, 3.0 equiv). The reaction was stirred at 110 °C for 2 h. The mixture was concentrated, dichloromethane (2.0 mL) was added, and the resulting mixture was stirred for 1 h and filtered to give crude 5-(1-(difluoromethyl)-1H-pyrazol-4-yl)pyrimidine-2,4-diol as a yellow solid (67 mg, 0.29 mmol). LCMS-ESI (m / z): [M-H]- calculated 227.05; found 227.12.

[0793] Step 3: At 0 °C, DIEA (114 mg, 0.88 mmol, 3.0 equiv) was added to a solution of 5-(1-(difluoromethyl)-1H-pyrazol-4-yl)pyrimidine-2,4-diol (67 mg, 0.29 mmol, 1.0 equiv) in POCl3 (1.5 mL). The reaction was stirred at 100 °C for 1 h. The mixture was concentrated, the pH was adjusted to 7 - 8 with aqueous NaHCO3, and the mixture was extracted with EA (2 mL × 3). Purification by flash column chromatography, eluting with petroleum ether and ethyl acetate (3:1), gave 2,4-dichloro-5-(1-(difluoromethyl)-1H-pyrazol-4-yl)pyrimidine as a white solid (21 mg, 0.08 mmol, 27%).

[0794] Step 4: NaHCO3 (13 mg, 0.16 mmol, 2.0 equiv) was added to a solution of Int-007 (20 mg, 0.08 mmol, 1.0 equiv) in DMF (2.0 mL). The reaction was stirred at 100 °C for 0.5 h. 2,4-Dichloro-5-(1-(difluoromethyl)-1H-pyrazol-4-yl)pyrimidine (21 mg, 0.08 mmol, 1.0 equiv) was added and the reaction was stirred at 100 °C for 3 h. The mixture was purified by preparative HPLC to give Example 304 (2.0 mg, 4.2 umol, 5%). LCMS-ESI (m / z): [M-H]- calculated 474.08; found 474.30. 1 1H NMR (DMSO-d6): δ 12.09 (s, 1H), 9.26 (s, 1H), 8.97 (s, 1H), 8.55 (s, 1H), 8.22 (d, J = 7.1 Hz, 1H), 8.06 - 7.77 (m, 2H), 7.20 (s, 1H), 7.01 (s, 1H), 3.44 - 3.41 (m, 2H), 2.91 - 2.88 (m, 2H).

[0795] Test method: Test method B was used. Retention time = 2.26 minutes.

[0796] Synthesis of Example 305

[0797]

[0798] Step 1: Add 2-[2-(2-chloro-5-iodopyrimidin-4-yl)oxy-3-fluoro-4-pyridyl]-1,5,6,7-tetrahydropyrrolo[3,2-c]pyridin-4-one (20 mg, 41.18 μmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (10.38 mg, 49.42 μmol, 1.2 equiv), Pd(dppf)Cl2 (3.01 mg, 4.12 μmol, 0.1 equiv) and K2CO3 (17.08 mg, 123.55 μmol, 3.0 equiv) to a vial. Flush the vial with Ar by pumping, and then suspend the contents in 1,4-dioxane (2 mL) and H2O (0.2 mL). Heat the reaction mixture to 100 °C with stirring for 7 h. Then, cool the solution, dilute with EtOAc, wash with brine, dry over Na2SO4, and then concentrate. Dissolve the crude material in DMSO and purify by preparative HPLC to obtain Example 096 (0.2 mg, 0.45 μmol, 1.1%). LCMS-ESI (m / z): [M+H]+ calculated 442.1; found 442.1. 1 1H NMR (DMSO-d6): δ 12.06 (s, 1H), 8.72 (s, 1H), 8.17 (d, J = 5.2 Hz, 1H), 7.79 (t, J = 5.4 Hz, 1H), 7.17 (s, 1H), 7.01 - 6.98 (m, 1H), 6.43 - 6.40 (m, 1H), 4.25 (q, J = 2.9 Hz, 2H), 3.82 (t, J = 5.7 Hz, 2H), 3.42 (dt, J = 6.6, 2.4 Hz, 3H), 2.88 (t, J = 7.1 Hz, 2H).

[0799] Synthesis of Example 306

[0800]

[0801] Step 1: A mixture of 5-bromo-2,4-dimethoxypyrimidine (1.2 g, 5.48 mmol, 1.0 equiv), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole (2.31 g, 10.96 mmol, 2.0 equiv), K2CO3 (2.27 g, 16.44 mmol, 3.0 equiv), and Pd(dppf)Cl2·DCM (895 mg, 1.10 mmol, 0.2 equiv) in 1,4-dioxane (20 mL) and water (4 mL) was stirred at 100 °C under a nitrogen atmosphere for 16 h. The reaction was cooled to room temperature and concentrated. The residue was purified by silica gel chromatography (0% to 50% EtOAc in petroleum ether) to give 5-(2,4-dimethoxypyrimidin-5-yl)thiazole (0.2 g, 895.85 μmol, 16%). LCMS-ESI (m / z): [M+H]+ calculated 224.04; found 224.02.

[0802] Step 2: A mixture of 5-(2,4-dimethoxypyrimidin-5-yl)thiazole (0.19 g, 851.06 μmol, 1.0 equiv) and NaI (383 mg, 2.55 mmol, 3.0 equiv) in AcOH (2 mL) was stirred at 110 °C for 1 h. The mixture was filtered. The cake was washed with DCM and dried to give crude 5-thiazol-5-ylpyrimidine-2,4-diol (160 mg, 819.68 μmol). The crude product was used in the next step without further purification. LCMS-ESI (m / z): [M+H]+ calculated 196.01; found 196.00.

[0803] Step 3: DIEA (0.5 mL) was added to a suspension of 5-(thiazol-5-yl)pyrimidine-2,4-diol (160 mg, 819.68 μmol, 1.0 equiv) in POCl3 (3 mL), and the reaction was stirred at 110 °C for 1 h. The resulting mixture was quenched with saturated NaHCO3 (aqueous solution, 200 mL) and extracted with EtOAc (80 mL x 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (0% to 30% EtOAc in petroleum ether) to give 5-(2,4-dichloropyrimidin-5-yl)thiazole (120 mg, 517.04 μmol, 63%). LCMS-ESI (m / z): [M+H]+ calculated 231.94; found 231.91.

[0804] Step 4: To a solution of Int-007 (50 mg, 202.25 μmol, 1.0 equiv) and 5-(2,4-dichloropyrimidin-5-yl)thiazole (51 mg, 222.47 μmol, 1.1 equiv) in DMSO (3 mL) was added K2CO3 (56 mg, 404.49 μmol, 2.0 equiv) and sodium methanesulfinate (0.6 mg, 6.07 μmol, 0.03 equiv). The mixture was stirred at room temperature for 2 h and purified by preparative HPLC to give Example 306 (7.3 mg, 16.48 μmol, 8%). LCMS-ESI (m / z): [M+H]+ calcd 443.04; found 442.79. 1 1H NMR (400 MHz, DMSO-d6) δ 12.07 (s, 1H), 9.32 (d, J = 0.4 Hz, 1H), 9.29 (s, 1H), 8.68 (d, J = 0.4 Hz, 1H), 8.20 (d, J = 5.2 Hz, 1H), 7.83 (t, J = 5.6 Hz, 1H), 7.18 (s, 1H), 7.00 (t, J = 2.4 Hz, 1H), 3.44 - 3.40 (m, 2H), 2.88 (t, J = 6.8 Hz, 2H).

[0805] Test method: Test method A was used. Retention time = 2.17 min.

[0806] Synthesis of Example 307

[0807]

[0808] Step 1: To a solution of 2,6-dimethoxypyrimidine-4-carboxylic acid (2.0 g, 10.87 mmol, 1.0 equiv) and N,O-dimethylhydroxylamine hydrochloride (1.27 g, 13.04 mmol, 1.2 equiv) in DMF (50 mL) was added triethylamine (3.29 g, 32.61 mmol, 3.0 equiv) and HATU (5 g, 13.04 mmol, 1.2 equiv). The reaction mixture was stirred at room temperature overnight. The reaction was diluted with water (100 mL) and extracted with EtOAc (50 mL × 3). The combined layers were washed with brine (50 mL) and dried over Na2SO4. The mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (0 - 50% EtOAc in PE) to give N,2,6-trimethoxy-N-methylpyrimidine-4-carboxamide (2.0 g, 81%). LCMS-ESI (m / z): [M+H]+ calcd 228.09; found 228.07.

[0809] Step 2: To a solution of N,2,6-trimethoxy-N-methylpyrimidine-4-carboxamide (2.5 g, 11.01 mmol, 1.0 equiv) in THF (50 mL) was added dropwise diisobutylaluminum hydride (1 M, 33.03 mL, 3.0 equiv) at -78 °C, and the reaction mixture was stirred at -78 °C for 3 h. The reaction was diluted with water (100 mL) and extracted with EtOAc (50 mL × 3). The combined layers were washed with brine (50 mL) and dried over Na2SO4. The mixture was filtered and the filtrate was concentrated to give 2,6-dimethoxypyrimidine-4-carbaldehyde (2.0 g). The crude product was used in the next step without further purification. LCMS-ESI (m / z): [M+H]+ calcd 169.05; found 169.01.

[0810] Step 3: To a solution of 2,6-dimethoxypyrimidine-4-carbaldehyde (1.0 g, 5.95 mmol, 1.0 equiv) in MeOH (20 mL) was added tosylmethyl isocyanide (1.39 g, 7.14 mmol, 1.2 equiv) and potassium carbonate (985 mg, 7.14 mmol, 1.2 equiv), and the reaction mixture was stirred at 80 °C for 3 h. The mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (0 - 50% EtOAc in PE) to give 5-(2,6-dimethoxypyrimidin-4-yl)oxazole (800 mg, 65%). LCMS-ESI (m / z): [M+H]+ calcd 208.06; found 208.02.

[0811] Step 4: To a solution of 5-(2,6-dimethoxypyrimidin-4-yl)oxazole (600 mg, 2.9 mmol, 1.0 equiv) in HOAc (10 mL) was added sodium iodide (1.3 g, 8.7 mmol, 3.0 equiv), and the reaction mixture was stirred at 110 °C for 1 h. The mixture was filtered and the filter cake was washed with DCM. The solid was dried to give 6-(oxazol-5-yl)pyrimidine-2,4-diol (600 mg, 100%). The crude product was used in the next step without further purification. LCMS-ESI (m / z): [M+H]+ calcd 180.03; found 179.98.

[0812] Step 5: To a solution of 6-(oxazol-5-yl)pyrimidine-2,4-diol (200 mg, 1.12 mmol, 1.0 equiv) in POCl3 (5 mL) was added DIPEA (1.5 mL), and the reaction mixture was stirred at 110 °C for 1 h. The reaction mixture was cooled to room temperature and then added dropwise to cold saturated aqueous NaHCO3. The solution was extracted with EtOAc (100 mL×3), washed with brine (100 mL), and dried over Na2SO4. The mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (0-50% EtOAc in PE) to afford 5-(2,6-dichloropyrimidin-4-yl)oxazole (136 mg, 56%).

[0813] Step 6: To a solution of 5-(2,6-dichloropyrimidin-4-yl)oxazole (100 mg, 0.47 mmol, 1.0 equiv) and Int-007 (116 mg, 0.47 mmol, 1.0 equiv) in DMSO (3 mL) were added K2CO3 (130 mg, 0.94 mol, 2.0 equiv) and sodium methylsulfinate (1.5 mg, 14 μmol, 0.03 equiv), and the reaction mixture was stirred at room temperature for 2 h. The mixture was filtered and the filtrate was purified by preparative HPLC to afford Example 307 (10.4 mg, 5%). LCMS-ESI (m / z): [M+H]+ calculated 427.06; found 427.20. 1 1H NMR (DMSO-d6): δ 11.96 (s, 1H), 8.81 (s, 1H), 8.59 (s, 1H), 8.25 (s, 1H), 8.03 (dd, J = 8.0 Hz, 1.2 Hz, 1H), 7.22 (m, 1H), 7.10 (m, 1H), 6.93 (m, 1H), 3.45 - 3.41 (m, 2H), 2.90 (t, J = 6.8 Hz, 2H).

[0814] Test method: Test method A used. Retention time = 2.07 min.

[0815] Synthesis of Example 308

[0816]

[0817] Step 1: A mixture of tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-1-carboxylate (1.48 g, 5.02 mmol, 2.2 equiv), 5-bromo-2,4-dimethoxypyrimidine (2) (500 mg, 2.28 mmol, 1.0 equiv), K2CO3 (946 mg, 6.85 mmol, 3.0 equiv.), and Pd(dppf)Cl2·DCM (186 mg, 228 μmol, 0.1 equiv) in 1,4-dioxane (25 mL) and water (2.5 mL) was stirred overnight at 90 °C under nitrogen. The mixture was concentrated and the residue was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (2:1 to 0:1), to give 2,4-dimethoxy-5-(1H-pyrazol-4-yl)pyrimidine (280 mg, 1.36 mmol, 59%). LCMS-ESI (m / z): [M+H]+ calculated 207.09; found 207.05.

[0818] Step 2: A mixture of 2,4-dimethoxy-5-(1H-pyrazol-4-yl)pyrimidine (124 mg, 601 μmol, 1.0 equiv) and NaI (283 mg, 1.89 mmol, 3.0 equiv) in HOAc (5 mL) was stirred at 110 °C for 1 h. The mixture was concentrated and the residue was diluted with DCM (10 mL). The mixture was stirred for 10 min and filtered. The filter cake was washed with DCM and dried to give crude 5-(1H-pyrazol-4-yl)pyrimidine-2,4-diol as a light pink solid (242 mg, containing NaI, NaOAc, HOAc, etc.). The crude product was used in the next step without further purification. LCMS-ESI (m / z): [M+H]+ calculated 179.06; found 179.07.

[0819] Step 3: DIEA (1 mL) was added to a suspension of 5-(1H-pyrazol-4-yl)pyrimidine-2,4-diol (222 mg crude, 1.0 equiv) in POCl3 (5 mL), and the reaction was stirred at 100 °C for 2 h. The mixture was concentrated and quenched with saturated NaHCO3 (aqueous solution, 50 mL), and extracted with EtOAc (50 mL × 4). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated in vacuo. The residue was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (4:1), to give 1-(4-(2,4-dichloropyrimidin-5-yl)-1H-pyrazol-1-yl)ethan-1-one (53 mg, 0.206 mmol, 37%). LCMS-ESI (m / z): [M+H]+ calculated 257.00; found 257.06.

[0820] Step 4: A mixture of Int-007 (52 mg, 210 μmol, 1.2 equiv) and NaHCO3 (53 mg, 631 μmol, 3.6 equiv) in DMF (3 mL) was stirred at 100 °C for 30 min, then a solution of 1-(4-(2,4-dichloropyrimidin-5-yl)-1H-pyrazol-1-yl)ethan-1-one (45 mg, 175 μmol, 1.0 equiv) in DMF (1 mL) was added, and the mixture was stirred at 100 °C for 2 h. The mixture was purified by preparative HPLC to give Example 308 (2 mg, 4.70 μmol, 2%). LCMS-ESI (m / z): [M+H]+ calculated 426.09; found 426.06. 1 1H NMR (400 MHz, DMSO-d6) δ 13.36 (s, 1H), 12.10 (s, 1H), 9.17 (s, 1H), 8.46 (s, 1H), 8.20 (d, J = 5.3 Hz, 2H), 7.83 (t, J = 5.4 Hz, 1H), 7.19 (s, 1H), 7.00 (s, 1H), 3.46 - 3.41 (m, 2H), 2.89 (t, J = 6.8 Hz, 2H).

[0821] Test method: Test method A was used. Retention time = 2.003 min.

[0822] Synthesis of Example 309

[0823]

[0824] Step 1: 5-Bromo-2,4-dimethoxypyrimidine (590 mg, 2.7 mmol, 1.0 equiv), 1,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (1.2 g, 5.4 mmol, 2.0 equiv), Pd(dppf)Cl2·CH2Cl2 (219 mg, 0.27 mmol, 0.1 equiv) and K2CO3 (1.12 g, 8.1 mmol, 3 equiv) in 1,4-dioxane (10 mL) and water (1 mL) were added to a round-bottom flask, and the reaction was stirred at 90 °C for 16 h under a nitrogen atmosphere. The mixture was filtered, the filtrate was concentrated and purified by silica gel column (0 - 4% MeOH in DCM) to give 5-(1,5-dimethyl-1H-pyrazol-4-yl)-2,4-dimethoxypyrimidine (600 mg, 94%). LCMS-ESI (m / z): [M+H]+ calculated 235.11; found 235.06.

[0825] Step 2: To a solution of 5-(1,5-dimethyl-1H-pyrazol-4-yl)-2,4-dimethoxypyrimidine (600 mg, 2.56 mmol, 1.0 equiv) in AcOH (20 mL) was added NaI (1.15 g, 7.68 mmol, 3 equiv), and the mixture was refluxed at 110 °C for 2 h. The solvent was evaporated, and the residue was added to saturated NaHCO3 solution (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (50 mL×2), dried over Na2SO4 and concentrated to give crude 5-(1,5-dimethyl-1H-pyrazol-4-yl)pyrimidine-2,4-diol (600 mg, 110%), which was used directly in the next step without further purification. LCMS-ESI (m / z): [M+H]+ calculated 207.08; found 206.81.

[0826] Step 3: To a solution of 5-(1,5-dimethyl-1H-pyrazol-4-yl)pyrimidine-2,4-diol (600 mg, 2.9 mmol, 1.0 equiv) in POCl3 (10 mL) was added DIEA (1.87 g, 14.5 mmol, 5.0 equiv), and the mixture was refluxed at 110 °C for 2 h. Then, most of the solvent was evaporated, and the residue was added dropwise to saturated NaHCO3 solution (50 mL), and the mixture was extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine (100 mL) and dried over Na2SO4, and the solvent was evaporated in vacuo and purified by silica gel column (0 - 30% EtOAc in petroleum ether) to give 2,4-dichloro-5-(1,5-dimethyl-1H-pyrazol-4-yl)pyrimidine as a yellow solid (280 mg, 40%).

[0827] LCMS-ESI (m / z): [M+H]+ calculated 243.01; found 242.97.

[0828] Step 4: To a solution of 2,4-dichloro-5-(1,5-dimethyl-1H-pyrazol-4-yl)pyrimidine (170 mg, 0.7 mmol, 1.0 equiv) in anhydrous DMSO (2 mL) was added 3-fluoro-4-iodopyridin-2-ol (184 mg, 0.77 mmol, 1.1 equiv), NaMeSO2 (2 mg, 0.02 mg, 0.03 equiv), and K2CO3 (193 mg, 1.4 mmol, 2 equiv). The mixture was stirred at room temperature for 2 h, then water (2 mL) was added. The solid was filtered and washed with water (10 mL). The solid was purified by silica gel column (0 - 70% EtOAc in petroleum ether) to give 2-chloro-5-(1,5-dimethyl-1H-pyrazol-4-yl)-4-((3-fluoro-4-iodopyridin-2-yl)oxy)pyrimidine (130 mg, 41%). LCMS-ESI (m / z): [M+H]+ calcd 445.96; found 445.88.

[0829] Step 5: To a round-bottom flask was added 2-chloro-5-(1,5-dimethyl-1H-pyrazol-4-yl)-4-((3-fluoro-4-iodopyridin-2-yl)oxy)pyrimidine (110 mg, 0.25 mmol, 1.0 equiv), Compound 1-6 (97 mg, 0.37 mmol, 1.5 equiv), Pd(dppf)Cl2·CH2Cl2 (20 mg, 0.025 mmol, 0.1 equiv), and K3PO4 (157 mg, 0.75 mmol, 3 equiv) in 1,4-dioxane (4 mL) and water (0.4 mL). The reaction was stirred at 70 °C for 16 h under a nitrogen atmosphere. The reaction mixture was purified by preparative HPLC to give Example 309 (5 mg, 4%). LCMS-ESI (m / z): [M+H]+ calcd 454.11; found 453.90. 1 H NMR (400 MHz, DMSO-d6) δ 12.04 (s, 1H), 8.73 (s, 1H), 8.17 (d, J = 5.2 Hz, 1H), 7.79 (t, J = 5.2 Hz, 1H), 7.67 (s, 1H), 7.18 (s, 1H), 7.00 (s, 1H), 3.82 (s, 3H), 3.43 (dt, J = 6.8, 2.4 Hz, 2H), 2.89 (t, J = 6.8 Hz, 2H), 2.37 (s, 3H).

[0830] Test method: Test method A was used. Retention time = 2.12 minutes.

[0831] Synthesis of Example 310

[0832]

[0833] Step 1: To a solution of 2-(2,4-dichloropyrimidin-5-yl)oxazole Int-0019 (200 mg, 1.02 mmol, 1.0 equiv) in DMF (10 mL) was added 3-fluoro-4-iodopyridin-2-ol (243 mg, 1.02 mmol, 1.0 equiv) and NaHCO3 (171 mg, 2.04 mol, 2.0 equiv), and then the reaction was stirred at 100 °C for 2 h. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and concentrated. The residue was purified by column chromatography (0 - 15% EtOAc in PE) to give 2-(2-chloro-4-((3-fluoro-4-iodopyridin-2-yl)oxy)pyrimidin-5-yl)oxazole (100 mg, 24%). LCMS-ESI (m / z): [M+H]+ calculated 418.91; found 418.97.

[0834] Step 2: To a solution of 2-(2-chloro-4-((3-fluoro-4-iodopyridin-2-yl)oxy)pyrimidin-5-yl)oxazole (100 mg, 0.24 mmol, 1.0 equiv) and 1-6 (126 mg, 0.48 mmol, 2.0 equiv) in 1,4-dioxane (10 mL) and water (2 mL) was added Pd(dppf)Cl2·DCM (20 mg, 24 μmol, 0.1 equiv) and NaHCO3 (60 mg, 0.72 mmol, 3.0 equiv), and the reaction was stirred at 65 °C for 1 h using a microwave reactor. The mixture was filtered and the filtrate was concentrated. The residue was purified by preparative HPLC to give Example 310 (2.1 mg, 2%). LCMS-ESI (m / z): [M+H]+ calculated 426.06; found 426.99. 1 1H NMR (DMSO-d6): δ 12.08 (s, 1H), 9.31 (s, 1H), 8.43 (d, J = 0.4 Hz, 1H), 8.18 (d, J = 5.2 Hz, 1H), 7.81 (t, J = 5.2 Hz, 1H), 7.56 (d, J = 0.8 Hz, 1H), 7.18 (s, 1H), 7.01 (s, 1H), 3.44 - 3.38 (m, 2H), 2.89 (t, J = 6.8 Hz, 2H).

[0835] Test method: Test method A used. Retention time = 2.04 min.

[0836] Retention time = 2.337 min.

[0837] Synthesis of Example 312

[0838]

[0839] Step 1: To a solution of methyl 2,4-dichloropyrimidine-5-carboxylate (1.0 g, 4.85 mmol, 1.0 equiv) in MeOH (15 mL) at 0 °C was added sodium methoxide (786 mg, 14.55 mmol, 3.0 equiv), and then the reaction was stirred at 0 °C for 2 h. The solution was concentrated. The residue was purified by column chromatography (0 - 33% EtOAc in PE) to give methyl 2,4-dimethoxypyrimidine-5-carboxylate (970 mg, 100%). LCMS-ESI (m / z): [M+H]+ calculated 199.06; found 199.02.

[0840] Step 2: To a solution of methyl 2,4-dimethoxypyrimidine-5-carboxylate (970 mg, 4.9 mmol, 1.0 equiv) in THF (10 mL) and water (10 mL) was added LiOH·H2O (412 mg, 9.8 mmol, 2.0 equiv), and then the reaction was stirred at room temperature for 1 h. The pH of the solution was adjusted to 3 with 1 M HCl, and then the mixture was extracted with EtOAc (20 mL × 3). The combined layers were washed with brine (50 mL), dried over Na2SO4. The mixture was filtered, and the filtrate was concentrated to give 2,4-dimethoxypyrimidine-5-carboxylic acid (930 mg, crude). LCMS-ESI (m / z): [M-H]- calculated 183.05; found 182.98.

[0841] Step 3: To a solution of 2,4-dimethoxypyrimidine-5-carboxylic acid (930 mg, 5.05 mmol, 1.0 equiv) and N,O-dimethylhydroxylamine hydrochloride (591 mg, 6.06 mmol, 1.2 equiv) in DMF (20 mL) were added triethylamine (2 mL, 15.15 mmol, 3.0 equiv) and HATU (2.3 g, 6.06 mmol, 1.2 equiv), and the reaction mixture was stirred at room temperature overnight. The reaction was diluted with water (100 mL) and extracted with EtOAc (50 mL × 3). The combined layers were washed with brine (50 mL), dried over Na2SO4. The mixture was filtered, and the filtrate was concentrated. The residue was purified by column chromatography (0 - 50% EtOAc in PE) to give N,2,4-trimethoxy-N-methylpyrimidine-5-carboxamide (960 mg, 84%).

[0842] LCMS-ESI (m / z): [M+H]+ calculated 228.09; found 228.07.

[0843] Step 4: Diisobutylaluminum hydride (1 M, 12.69 mL, 12.69 mmol, 3.0 equiv) was added dropwise to a solution of N,2,4-trimethoxy-N-methylpyrimidine-5-carboxamide (960 mg, 4.23 mmol, 1.0 equiv) in THF (20 mL) at -78 °C. The solution was stirred at -78 °C for 3 h. The reaction was diluted with water (50 mL) and extracted with EtOAc (50 mL × 3). The combined layers were washed with brine (50 mL), dried over Na2SO4. The mixture was filtered and the filtrate was concentrated to give 2,4-dimethoxypyrimidine-5-carbaldehyde (640 mg, 91%). LCMS-ESI (m / z): [M+H]+ calcd 169.05; found 169.03.

[0844] Step 5e: 1-((1-Isocyanatoethyl)sulfonyl)-4-methylbenzene (951 mg, 4.55 mmol, 1.2 equiv) and potassium carbonate (628 mg, 4.55 mmol, 1.2 equiv) were added to a solution of 2,4-dimethoxypyrimidine-5-carbaldehyde (640 mg, 3.79 mmol, 1.0 equiv) in MeOH (20 mL). The reaction was stirred at 80 °C for 3 h. The mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (0 - 50% EtOAc in PE) to give 5-(2,4-dimethoxypyrimidin-5-yl)-4-methyloxazole (440 mg, 52%). LCMS-ESI (m / z): [M+H]+ calcd 222.08; found 221.98.

[0845] Step 6l: Sodium iodide (900 mg, 6.0 mmol, 3.0 equiv) was added to a solution of 5-(2,4-dimethoxypyrimidin-5-yl)-4-methyloxazole (440 mg, 2.0 mmol, 1.0 equiv) in HOAc (10 mL). The reaction was stirred at 110 °C for 1 h. The mixture was filtered and the filter cake was washed with DCM. The solid was dried to give 5-(4-methyloxazol-5-yl)pyrimidine-2,4-diol (400 mg, 100%). LCMS-ESI (m / z): [M+H]+ calcd 194.05; found 194.02.

[0846] Step 7: To a solution of 5-(4-methyl-oxazol-5-yl)pyrimidine-2,4-diol (400 mg, 2.07 mmol, 1.0 eq) in POCl3 (5 mL) was added DIPEA (1.5 mL), and the reaction was stirred at 110 °C for 1 h. The reaction mixture was cooled to room temperature, and then the reaction mixture was added dropwise to cold saturated NaHCO3 solution. The solution was extracted with EtOAc (100 mL × 3), washed with brine (100 mL), and dried over Na2SO4. The mixture was filtered, and the filtrate was concentrated. The residue was purified by column chromatography (0 - 33% EtOAc in PE) to give 5-(2,4-dichloropyrimidin-5-yl)-4-methyloxazole (250 mg, 53%).

[0847] Step 8: To a solution of 5-(2,4-dichloropyrimidin-5-yl)-4-methyloxazole (120 mg, 0.52 mmol, 1.0 eq) and Int-007 (128 mg, 0.52 mmol, 1.0 eq) in DMSO (3 mL) were added K2CO3 (144 mg, 1.04 mmol, 2.0 eq) and sodium methylsulfinate (1.6 mg, 16 μmol, 0.03 eq), and the reaction mixture was stirred at 50 °C for 2 h. The mixture was filtered, and the filtrate was purified by preparative HPLC to give Example 312 (6.6 mg, 3%). LCMS-ESI (m / z): [M+H]+ calculated 441.08; found 440.93. 1 1H NMR (DMSO-d6): δ 12.07 (s, 1H), 8.96 (s, 1H), 8.56 (s, 1H), 8.19 (d, J = 5.6 Hz, 1H), 7.81 (m, 1H), 7.18 (s, 1H), 7.00 (s, 1H), 3.44 - 3.39 (m, 2H), 2.89 (t, J = 6.8 Hz, 2H), 2.31 (s, 3H).

[0848] Test method: Test method A was used. Retention time = 2.07 min.

[0849] Synthesis of Example 313

[0850]

[0851] Step 1: A mixture of 5-iodo-2,4-dimethoxypyrimidine (600 mg, 2.25 mmol, 1.0 equiv), 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)oxazole (470 mg, 2.25 mmol, 1.0 equiv), cesium carbonate (2.2 g, 6.75 mmol, 3.0 equiv) and Pd(dppf)Cl2·CH2Cl2 (184 mg, 225 μmol, 0.1 equiv) in 1,4-dioxane (50 mL) and water (10 mL) was stirred at 120 °C overnight. The mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (0 - 20% EtOAc in PE) to give 5-(2,4-dimethoxypyrimidin-5-yl)-2-methyloxazole (330 mg, 66%). LCMS-ESI (m / z): [M+H]+ calcd 222.08; found 221.99.

[0852] Step 2: Sodium iodide (671 mg, 4.47 mmol, 3.0 equiv) was added to a solution of 5-(2,4-dimethoxypyrimidin-5-yl)-2-methyloxazole (330 mg, 1.49 mmol, 1.0 equiv) in HOAc (10 mL), and the reaction was stirred at 110 °C for 1 h. The mixture was filtered and the cake was washed with DCM. The solid was dried to give 5-(2-methyloxazol-5-yl)pyrimidine-2,4-diol (200 mg, 69%). LCMS-ESI (m / z): [M+H]+ calcd 194.05; found 194.02.

[0853] Step 3: DIPEA (1.5 mL) was added to a solution of 5-(2-methyloxazol-5-yl)pyrimidine-2,4-diol (200 mg, 1.04 mmol, 1.0 equiv) in POCl3 (5 mL), and the reaction was stirred at 110 °C for 1 h. The reaction mixture was cooled to room temperature and then the reaction mixture was added dropwise to cold saturated NaHCO3 solution. The solution was extracted with EtOAc (100 mL×3), washed with brine (100 mL), dried over Na2SO4 and concentrated. The residue was purified by column chromatography (0 - 33% EtOAc in PE) to give 5-(2,4-dichloropyrimidin-5-yl)-2-methyloxazole (120 mg, 51%).

[0854] Step 4: To a solution of 5-(2,4-dichloropyrimidin-5-yl)-2-methyloxazole (120 mg, 0.52 mmol, 1.0 eq) and Int-007 (128 mg, 0.52 mmol, 1.0 eq) in DMSO (3 mL) was added K2CO3 (144 mg, 1.04 mmol, 2.0 eq) and sodium methylsulfinate (1.6 mg, 16 μmol, 0.03 eq). The reaction mixture was stirred at 50 °C for 2 h. The mixture was filtered and the filtrate was purified by preparative HPLC to give Example 313 (15 mg, 7%). LCMS-ESI (m / z): [M+H]+ calculated 441.08; found 440.94. 1 1H NMR (DMSO-d6): δ 12.08 (s, 1H), 9.05 (s, 1H), 8.22 (d, J = 5.2 Hz, 1H), 8.45 (m, 1H), 7.74 (s, 1H), 7.19 (s, 1H), 7.00 (s, 1H), 3.45 - 3.41 (m, 2H), 2.90 (t, J = 6.8 Hz, 2H), 2.56 (s, 3H).

[0855] Test method: Test method A was used. Retention time = 2.07 min.

[0856] Synthesis of Example 314

[0857]

[0858] Step 1: A mixture of 2,4-dimethyloxazole (1 g, 10.30 mmol, 1.0 eq), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (2.6 g, 10.30 mmol, 1.0 eq), 4,4'-di-tert-butyl-2,2'-bipyridine (165 mg, 617.82 μmol, 0.06 eq) and (1,5-cyclooctadiene)(methoxy)iridium(I) dimer (204 mg, 308.91 μmol, 0.03 eq) in THF (20 mL) was stirred at 50 °C for 16 h under a nitrogen atmosphere. The reaction was cooled to room temperature and concentrated under reduced pressure to give a residue. The residue was purified by silica gel chromatography using a gradient of 0% to 30% EtOAc in petroleum ether to give 2,4-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)oxazole (1.5 g, 6.72 mmol, 65%). LCMS-ESI (m / z): [M+H]+ calculated 224.14; found 224.13.

[0859] Step 2: A mixture of 4-[(4-bromo-2-pyridyl)oxy]-2-chloro-5-iodopyrimidine (500 mg, 1.21 mmol, 1.0 equiv), 2,4-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)oxazole (405 mg, 1.82 mmol, 1.5 equiv), K3PO4 (772 mg, 3.64 mmol, 3.0 equiv), and Pd(dppf)Cl2·DCM (150 mg, 181.86 μmol, 0.15 equiv) in 1,4-dioxane (10 mL) and water (2 mL) was stirred at 55 °C for 16 h under a nitrogen atmosphere. The reaction was cooled to room temperature and concentrated under reduced pressure to give a residue. The residue was purified by silica gel chromatography using a gradient of 0% to 50% EtOAc in petroleum ether to afford 5-[4-[(4-bromo-2-pyridyl)oxy]-2-chloropyrimidin-5-yl]-2,4-dimethyloxazole as a red solid (420 mg, 1.10 mmol, 90%). LCMS-ESI (m / z): [M+H]+ calcd 380.97; found 382.76.

[0860] Step 3: A mixture of 5-[4-[(4-bromo-2-pyridyl)oxy]-2-chloropyrimidin-5-yl]-2,4-dimethyloxazole (100 mg, 262.05 μmol, 1.0 equiv), 1-6 (82 mg, 314.46 μmol, 1.2 equiv), K3PO4 (167 mg, 786.14 μmol, 3.0 equiv), and Pd(dppf)Cl2·DCM (22 mg, 26.20 μmol, 0.1 equiv) in 1,4-dioxane (5 mL) and water (1 mL) was stirred in a microwave reactor at 100 °C for 2 h under a nitrogen atmosphere. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC to afford Example 314 (6.6 mg, 15.11 μmol, 5.8%). LCMS-ESI (m / z): [M+H]+ calcd 437.11; found 436.94. 1 H NMR (400 MHz, DMSO-d6) δ 11.99 (s, 1H), 8.87 (s, 1H), 8.28 (d, J = 5.2 Hz, 1H), 7.64 - 7.62 (m, 1H), 7.57 - 7.56 (m, 1H), 7.14 - 7.11 (m, 2H), 3.44 - 3.39 (m, 2H), 2.85 (t, J = 6.8 Hz, 2H), 2.44 (s, 3H), 2.23 (s, 3H).

[0861] Test method: Test method A was used. Retention time = 2.11 minutes.

[0862] Synthesis of Example 315

[0863]

[0864] Step 1: A mixture of 2,4-dimethyloxazole (1 g, 10.30 mmol, 1.0 equiv), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (2.6 g, 10.30 mmol, 1.0 equiv), 4,4'-di-tert-butyl-2,2'-bipyridine (165 mg, 617.82 μmol, 0.06 equiv) and (1,5-cyclooctadiene)(methoxy)iridium(I) dimer (204 mg, 308.91 μmol, 0.03 equiv) in THF (20 mL) was stirred at 50 °C under a nitrogen atmosphere for 16 h. The reaction was cooled to room temperature and concentrated to give a residue. The residue was purified by silica gel chromatography using a gradient of 0% to 30% EtOAc in petroleum ether to give 2,4-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)oxazole (1.5 g, 6.72 mmol, 65%). LCMS-ESI (m / z): [M+H]+ calculated 224.14; found 224.13.

[0865] Step 2: A mixture of 5-bromo-2,4-dimethoxypyrimidine (1 g, 4.57 mmol, 1.0 equiv), 2,4-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)oxazole (1.5 g, 6.85 mmol, 2.0 equiv), K2CO3 (1.89 g, 13.70 mmol, 3.0 equiv), Pd(dppf)Cl2·DCM (372 mg, 456.55 μmol, 0.1 equiv) in 1,4-dioxane (20 mL) and water (4 mL) was stirred at 100 °C under a nitrogen atmosphere for 16 h. The reaction was cooled to room temperature and concentrated to give a residue. The residue was purified by silica gel chromatography using a gradient of 0% to 50% EtOAc in petroleum ether to give 5-(2,4-dimethoxypyrimidin-5-yl)-2,4-dimethyloxazole (0.7 g, 2.98 mmol, 65%). LCMS-ESI (m / z): [M+H]+ calculated 236.10; found 236.05.

[0866] Step 3: A mixture of 5-(2,4-dimethoxypyrimidin-5-yl)-2,4-dimethyloxazole (0.7 g, 2.98 mmol, 1.0 equiv) and NaI (1.34 g, 8.93 mmol, 3.0 equiv) in AcOH (7 mL) was stirred at 110 °C for 1 h. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by silica gel chromatography using a gradient of 0% to 10% MeOH in DCM to give 5-(2,4-dimethyloxazol-5-yl)pyrimidine-2,4-diol (0.5 g, 2.41 mmol, 81%). LCMS-ESI (m / z): [M+H]+ calcd 208.06; found 208.07.

[0867] Step 4: DIEA (0.5 mL) was added to a suspension of 5-(2,4-dimethyloxazol-5-yl)pyrimidine-2,4-diol (0.5 g, 2.41 mmol, 1.0 equiv) in POCl3 (4 mL), and the mixture was stirred at 110 °C for 1 h. The mixture was quenched with saturated NaHCO3 (aqueous solution, 400 mL) and extracted with EtOAc (150 mL x 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography using a gradient of 0% to 30% EtOAc in petroleum ether to give 5-(2,4-dichloropyrimidin-5-yl)-2,4-dimethyloxazole as a yellow solid (0.2 g, 819.41 μmol, 34%). LCMS-ESI (m / z): [M+H]+ calcd 244.00; found 243.97.

[0868] Step 5: K2CO3 (80 mg, 582.47 μmol, 2.0 equiv) and sodium methanesulfinate (891 μg, 8.74 μmol, 0.03 equiv) were added to a solution of Int-007 (80 mg, 291.23 μmol, 1.0 equiv) and 5-(2,4-dichloropyrimidin-5-yl)-2,4-dimethyloxazole (78 mg, 320.36 μmol, 1.1 equiv) in DMSO (3 mL), and the mixture was stirred at room temperature for 2 h. The mixture was purified by preparative HPLC to give Example 315 (6.1 mg, 13.41 μmol, 4.6%). LCMS-ESI (m / z): [M+H]+ calcd 455.10; found 454.96. 11H NMR (400 MHz, DMSO-d6) δ 12.05 (s, 1H), 8.91 (s, 1H), 8.18 (d, J = 5.2 Hz, 1H), 7.81 (t, J = 5.6 Hz, 1H), 7.18 (s, 1H), 7.00 (t, J = 2.4 Hz, 1H), 3.44 - 3.40 (m, 2H), 2.89 (t, J = 6.8 Hz, 2H), 2.45 (s, 3H), 2.24 (s, 3H).

[0869] Test method: Test method A used. Retention time = 2.237 minutes.

[0870] Synthesis of Example 316

[0871]

[0872] Step 1: To a solution of 2,6-dimethoxypyrimidine-4-carboxylic acid (1.00 g, 5.43 mmol, 1.0 equiv), HATU (2.5 g, 6.63 mmol, 1.22 equiv, turns yellow) in DMF (25 mL) was added Et3N (2.3 mL). The reaction mixture was stirred at room temperature for 2 h. The mixture was diluted with water (70 mL) and extracted with EtOAc (25 mL × 3), washed with aqueous NaCl solution, dried over Na2SO4 to give N,2,6-trimethoxy-N-methylpyrimidine-4-carboxamide (1.09 g, 89%). LCMS-ESI (m / z): [M+H]+ calculated 228.09; found 228.07.

[0873] Step 2: The mixture of N,2,6-trimethoxy-N-methylpyrimidine-4-carboxamide (1.09 g, 4.81 mmol, 1.0 equiv) in THF (20 mL) was cooled to -78 °C, then a solution of DIBAL-H (14.4 mL, 14.4 mmol, 3.0 equiv, 1 M in THF) was added dropwise to the reaction. The mixture was stirred at -78 °C for 4 h. The reaction was quenched with saturated aqueous H4Cl solution and extracted with EtOAc (50 mL × 3). The combined organic phases were filtered through Celite, and the filtrate was washed with aqueous NaCl solution. The organic phase was dried over Na2SO4 and then concentrated to give 2,6-dimethoxypyrimidine-4-carbaldehyde (793 mg, 4.72 mmol, 98%). The crude product was used in the next step without further purification.

[0874] Step 3: A mixture of 2,6-dimethoxypyrimidine-4-carbaldehyde (0.79 g, 4.72 mmol, 1.0 equiv), K2CO3 (0.98 g, 7.10 mmol, 1.5 equiv) and 2-tosylpropionitrile (1.19 g, 5.69 mmol, 1.2 equiv) in MeOH (20 mL) was stirred at 80 °C for 3 h. The mixture was concentrated and purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (2:1) to give 5-(2,6-dimethoxypyrimidin-4-yl)-4-methyloxazole (0.52 g, 49%). LCMS-ESI (m / z): [M+H]+ calcd 222.08; found 222.06.

[0875] Step 4: A mixture of 5-(2,6-dimethoxypyrimidin-4-yl)-4-methyloxazole (0.51 g, 2.33 mmol, 1.0 equiv) and NaI (1.05 g, 7.00 mmol, 3.0 equiv) in AcOH (5 mL) was stirred at 110 °C for 2 h. The solvent was evaporated and the residue was added to saturated NaHCO3 solution (30 mL). The mixture was extracted with EtOAc (30 mL×3), and the combined organic layers were washed with brine (50 mL×2), dried over Na2SO4 and concentrated to give 6-(4-methyloxazol-5-yl)pyrimidine-2,4-diol (564 mg). The crude product was used in the next step without further purification.

[0876] Step 5: To a mixture of 6-(4-methyloxazol-5-yl)pyrimidine-2,4-diol (0.564 g, crude, 1.0 equiv) in POCl3 (6 mL) was added DIPEA (2 mL), and the mixture was stirred at 110 °C for 1 h. The mixture was quenched slowly with saturated NaHCO3 (aqueous solution, 250 mL), extracted with EtOAc (150 mL×2), and the combined organic phases were washed with saturated NaCl aqueous solution, dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography, eluting with petroleum ether and EtOAc (4:1) to give 5-(2,6-dichloropyrimidin-4-yl)-4-methyloxazole (69 mg, 10%). LCMS-ESI (m / z): [M+H]+ calcd 229.98; found 229.99.

[0877] Step 6: A mixture of 5-(2,6-dichloropyrimidin-4-yl)-4-methyloxazole (69 mg, 0.30 mmol, 1.0 equiv), Int-007 (74 mg, 0.30 mmol, 1.0 equiv), K2CO3 (83 mg, 0.60 mmol, 2.0 equiv) and sodium methanesulfinate (1 mg, 0.01 mmol, 0.03 equiv) in DMSO (2 mL) was stirred at room temperature for 3 h. The mixture was purified by preparative HPLC to afford Example 316 as a yellow solid (8.1 mg, 6%). LCMS-ESI (m / z): [M+H]+ calcd 441.08; found 440.93. 1 1H NMR (400 MHz, DMSO-d6) δ 11.96 (s, 1H), 8.68 (s, 1H), 8.51 (s, 1H), 8.06 (d, J = 7.2 Hz, 1H), 7.23 (s, 1H), 7.10 (s, 1H), 6.93 (t, J = 7.2 Hz, 1H), 3.28 (td, J = 7.8, 2.4 Hz, 2H), 2.90 (t, J = 6.8 Hz, 2H), 2.60 (s, 3H).

[0878] Test method: Test method A used. Retention time = 2.323 min.

[0879] Synthesis of Example 317

[0880]

[0881] Step 1: 4-Bromo-2,6-dimethoxypyrimidine (500 mg, 2.28 mmol, 1.0 equiv), (1-methyl-1H-pyrazol-4-yl)boronic acid (575 mg, 4.56 mmol, 2.0 equiv), Pd(dppf)Cl2·CH2Cl2 (185 mg, 0.22 mmol, 0.1 equiv) and K2CO3 (946 mg, 6.84 mmol, 3 equiv) were added to a round-bottom flask, then 1,4-dioxane (10 mL) and water (1 mL) were added, and then the flask was flushed 3 times with vacuum and nitrogen. The mixture was stirred at 90 °C for 16 h under a nitrogen atmosphere. After the reaction, the solid was filtered off, the filtrate was concentrated in vacuo and purified by silica gel column (0 - 4% MeOH in DCM) to afford 2,4-dimethoxy-6-(1-methyl-1H-pyrazol-4-yl)pyrimidine (430 mg, 85%). LCMS-ESI (m / z): [M+H]+ calcd 221.10; found 221.06.

[0882] Step 2: To a solution of 2,4-dimethoxy-6-(1-methyl-1H-pyrazol-4-yl)pyrimidine (430 mg, 1.95 mmol, 1.0 equiv) in AcOH (10 mL) was added NaI (878 mg, 5.86 mmol, 3 equiv), and the mixture was refluxed at 110 °C for 2 h. The solvent was evaporated, and the residue was added to saturated NaHCO3 (30 mL) solution. The mixture was extracted with EtOAc (30 mL × 3), and the combined organic layers were washed with brine (50 mL × 2), dried over Na2SO4 and concentrated to give 6-(1-methyl-1H-pyrazol-4-yl)pyrimidine-2,4-diol (300 mg, 79%), which was used directly in the next step without further purification. LCMS-ESI (m / z): [M+H]+ calcd 193.06; found 193.04.

[0883] Step 3: To a solution of 6-(1-methyl-1H-pyrazol-4-yl)pyrimidine-2,4-diol (300 mg, 1.56 mmol, 1.0 equiv) in POCl3 (5 mL) was added DIEA (1.01 g, 7.81 mmol, 5.0 equiv), and the mixture was refluxed at 110 °C for 2 h. Then most of the solvent was evaporated in vacuo, and the residue was added dropwise to saturated NaHCO3 solution (50 mL), extracted with EtOAc (50 mL x3), and the combined organic layers were washed with brine (100 mL) and dried over Na2SO4. The solvent was evaporated in vacuo and purified by silica gel column (0-30% EtOAc in petroleum ether) to give 2,4-dichloro-6-(1-methyl-1H-pyrazol-4-yl)pyrimidine (320 mg, 68%). LCMS-ESI (m / z): [M+H]+ calcd 229.0; found 229.01.

[0884] Step 4: To a solution of Int-007 (78 mg, 0.32 mmol, 1.0 equiv) in anhydrous DMSO (1 mL) were added compound 3 (80 mg, 0.35 mmol, 1.1 equiv), MeSO2Na (0.9 mg, 0.009 mmol, 0.03 equiv) and K2CO3 (87 mg, 0.64 mmol, 2.0 equiv), and the mixture was stirred at room temperature for 2 h. After the reaction, water (2 mL) was added to the reaction mixture, and the solid was filtered and washed with water (20 mL). Then the solid was purified by preparative HPLC to give Example 317 as a light yellow solid (50 mg, 29%). LCMS-ESI (m / z): [M+H]+ calcd 440.10; found 440.26. 11H NMR (400 MHz, DMSO-d6) δ 12.07 (s, 1H), 9.15 (s, 1H), 8.43 (s, 1H), 8.21 (d, J = 5.2 Hz, 1H), 8.17 (s, 1H), 7.82 (t, J = 5.2 Hz, 1H), 7.19 (s, 1H), 7.00 (t, J = 5.2 Hz, 1H), 3.92 (s, 3H), 3.43 (dt, J = 6.8, 2.0 Hz, 2H), 2.89 (t, J = 6.8 Hz, 2H).

[0885] Test method: Test method A was used. Retention time = 2.08 minutes.

[0886] Synthesis of Example 318

[0887]

[0888] Step 1: To a solution of (2,4-dimethoxypyrimidin-5-yl)boronic acid (100 mg, 0.62 mmol, 1.0 equiv) and 4-bromo-1-methyl-1H-1,2,3-triazole (136 mg, 0.74 mmol, 1.2 equiv) in 1,4-dioxane (1 mL) and water (0.1 mL) was added Pd(dppf)Cl2·DCM (50 mg, 0.06 mmol, 0.10 equiv) and K2CO3 (170 mg, 1.23 mmol, 2.0 equiv). The mixture was stirred overnight at 90 °C under nitrogen and purified by flash column chromatography, eluting with dichloromethane and methanol (10:1) to give 2,4-dimethoxy-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrimidine (160 mg, 99%). LCMS-ESI (m / z): [M+H]+ calculated 222.10; found 222.06.

[0889] Step 2: To a solution of 2,4-dimethoxy-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrimidine (160 mg, 0.72 mmol, 1.0 equiv) in HOAc (3 mL) was added NaI (325 mg, 2.17 mmol, 3.0 equiv). The reaction was stirred at 110 °C for 2 h, the mixture was concentrated and dichloromethane (6 mL) was added. The mixture was stirred for 1 h and filtered to give 5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrimidine-2,4-diol (165 mg, 99%). LCMS-ESI (m / z): [M-H]+ calculated 194.07; found 194.01.

[0890] Step 3: At 0 °C, DIEA (551 g, 4.27 mmol, 5.0 equiv) was added to a solution of 5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrimidine-2,4-diol (165 mg, 0.85 mmol, 1.0 equiv) in POCl3 (1.5 mL), and the mixture was stirred at 110 °C for 1 h. The mixture was concentrated and the pH was adjusted to 7 - 8 with aqueous NaHCO3. The mixture was extracted with EA (10 mL × 3) and purified by flash column chromatography, eluting with dichloromethane and methanol (10:1) to give 2,4-dichloro-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrimidine (40 mg, 20%). LCMS-ESI (m / z): [M+H]+ calculated 230.00; found 229.98.

[0891] Step 4: To a solution of 2,4-dichloro-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrimidine (20 mg, 0.08 mmol, 1.0 equiv) in DMSO (1 mL) was added Int-003 (47 mg, 0.19 mmol, 1.1 equiv), CH3SO2Na (0.5 mg, 0.005 mmol, 0.03 equiv) and K2CO3 (48 mg, 0.34 mmol, 2.0 equiv). The reaction mixture was stirred at room temperature for 2 h and the mixture was purified by preparative HPLC to give Example 318 (8.1 mg, 10%). LCMS-ESI (m / z): [M+H]+ calculated 441.10; found 441.07. 1 1H NMR (DMSO-d6): δ 11.89 (s, 1H), 9.45 (s, 1H), 8.33 (s, 1H), 7.66 (d, J = 8.0 Hz, 1H), 7.20 (s, 1H), 7.07 (s, 1H), 6.89 (t, J = 7.2 Hz, 1H), 4.08 (s, 3H), 3.43 (d, J = 2.4 Hz, 2H), 2.90 (t, J = 6.8 Hz, 2H).

[0892] Test method: Test method A was used. Retention time = 2.12 min.

[0893] Synthesis of Example 319

[0894]

[0895] Step 1: To a solution of (2,4-dichloropyrimidin-5-yl)methanol (200 mg, 1.12 mmol, 1.0 eq) in DCM (5 mL) was added tert-butyl 2,2,2-trichloroacetimidate (268 mg, 1.23 mmol, 1.1 eq) and boron trifluoride diethyl etherate (10 mL); the mixture was stirred overnight at room temperature. The reaction mixture was concentrated and purified by silica gel column chromatography (0 - 10% EtOAc in petroleum ether) to give 5-(tert-butoxymethyl)-2,4-dichloropyrimidine as a colorless oil (70 mg, 26%). LCMS-ESI (m / z): [M+H]+ calculated 235.03; found 235.02.

[0896] Step 2: To a solution of Int-007 (67 mg, 0.27 mmol, 1.0 eq) in anhydrous DMSO (1 mL) was added 5-(tert-butoxymethyl)-2,4-dichloropyrimidine (70 mg, 0.30 mmol, 1.1 eq), NaMeSO2 (0.8 mg, 0.008 mmol, 0.03 eq), and K2CO3 (75 mg, 0.64 mmol, 2 eq), and the mixture was stirred at room temperature for 2 h. The reaction was diluted with water (2 mL) and filtered, and the solid was washed with water (20 mL) and purified by preparative HPLC to give Example 319 as a light yellow solid (4 mg, 3%). LCMS-ESI (m / z): [M+H]+ calculated 446.13; found 446.14. 1 1H NMR (400 MHz, DMSO-d6) δ 12.05 (s, 1H), 8.74 (s, 1H), 8.19 (d, J = 5.6 Hz, 1H), 7.80 (t, J = 5.6 Hz, 1H), 7.19 (s, 1H), 7.00 (t, J = 2.4 Hz, 1H), 4.59 (s, 2H), 3.42 (dt, J = 6.8, 2.4 Hz, 2H), 2.89 (t, J = 6.8 Hz, 2H), 1.26 (s, 9H).

[0897] Test method: Test method A was used. Retention time = 2.48 min.

[0898] Synthesis of Example 320

[0899]

[0900] Step 1: At 0 °C, 3-(tert-butoxycarbonylamino)-2-methyl-propanoic acid (900 mg, 4.4 mmol) and N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (1.44 g, 7.5 mmol) were added to a stirred solution of 2,2-dimethyl-1,3-dioxane-4,6-dione (702 mg, 4.8 mmol) and DMAP (811 mg, 6.6 mmol) in dichloromethane (15 mL). The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was diluted with dichloromethane (80 mL) and washed with cold 5% KHSO4 (75 mL × 3) and brine (75 mL). The organic layer was dried over MgSO4 and filtered. The filtrate was concentrated in vacuo to give tert-butyl N-[3-(2,2-dimethyl-4,6-dioxo-1,3-dioxan-5-yl)-2-methyl-3-oxo-propyl]carbamate (1.1 g, 75%). LCMS-ESI (m / z): [M+H]+ calcd 330.15; found 330.06.

[0901] Step 2: A solution of tert-butyl N-[3-(2,2-dimethyl-4,6-dioxo-1,3-dioxan-5-yl)-2-methyl-3-oxo-propyl]carbamate (1.1 g, 3.3 mmol) in EtOAc (30 mL) was stirred at 85 °C overnight under a nitrogen atmosphere. The reaction was concentrated and purified by silica gel column chromatography using a gradient of 0 to 30% EtOAc in petroleum ether to give tert-butyl 5-methyl-2,4-dioxo-piperidine-1-carboxylate (600 mg, 62%). LCMS-ESI (m / z): [M+H]+ calcd 228.12; found 228.06.

[0902] Step 3: A mixture of 2-bromo-1-(3-bromo-2-fluorophenyl)ethanone (373 mg, 990 μmol), tert-butyl 5-methyl-2,4-dioxo-piperidine-1-carboxylate (250 mg, 1.1 mmol) and ammonium acetate (339 mg, 4.4 mmol) in EtOH (1 mL) was stirred at 60 °C for 2 h. The mixture was concentrated and purified by silica gel column chromatography using a gradient of 0 - 3% MeOH in DCM to give tert-butyl 2-(2-bromo-3-fluoropyridin-4-yl)-7-methyl-4-oxo-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridine-5-car...

Claims

1. A compound of formula (I): or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein, X1, X2, X3 and X4 are each independently selected from a bond, CH, N, NH, O or S; Y1, Y2, Y3, Y4, Y5 and Y6 are each independently selected from a bond, CH, N, O or S; M1, M2, M3, M4, M5 and M6 are each independently selected from a bond, CH, N, O or S; L is a bond, NH, O, S, alkyl, alkoxy, alkyl-N- or -CONH-; The conditions are that a) when L is -CONH-, it is a 5-membered ring; b) when L is a bond, it is a 5-membered ring; R1, R2, R3, R4 and R5 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, hydroxyalkyl, cyano-alkyl, oxo, -CONH2, alkenyl-C(O)NH-, alkynyl-NH-, -C(O)O-alkyl, -C(O)NH-alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein each of said alkyl, haloalkyl, hydroxyalkyl, oxo, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently unsubstituted or substituted at each occurrence by one or more substituents selected from deuterium, halogen, amino, nitro, cyano, hydroxy, alkyl, alkoxy, haloalkyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; or R2 and R3 together with the C atom to which they are attached form a heterocyclyl, wherein said heterocyclyl is optionally substituted by one or more substituents selected from halogen, amino, nitro, cyano, hydroxy, alkyl, alkoxy, haloalkyl, hydroxyalkyl, cycloalkyl and heterocyclyl; t is 0, 1, 2 or 3; and s 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 deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein, L is a key, NH, O, S, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkyl-N- or -CONH-; Preferably, L is a bond, NH, O, S, C 1-3 alkyl, C 1-3 alkoxy, C 1-3 alkyl-N- or -CONH-.

3. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein, R1 is selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group, -CONH2; Preferably, R1 is selected from hydrogen, deuterium, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl.

4. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein, R2 and R3 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group, -CONH2; Preferably, R2 and R3 are each independently selected from hydrogen, deuterium, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl; alternatively, R2 and R3 together with the C atom to which they are attached form a 4- to 6-membered heterocyclyl containing 1, 2 or 3 heteroatoms selected from N, O or S; Preferably, R2 and R3 together with the C atom to which they are attached form 5. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein, R4 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group, -CONH2; Preferably, R4 is independently selected from hydrogen, deuterium, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, amino, F, Cl or Br.

6. The compound according to claim 1, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein, R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 2-6 cyano-alkyl, oxo, -CONH2, C 3-6 alkenyl-C(O)NH-, C 2-6 alkynyl-NH-, -C(O)O-C 1-4 alkyl, -C(O)NH-C 1-4 alkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 6-10 aryl, a 5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O, wherein each of said alkyl, haloalkyl, hydroxyalkyl, cycloalkyl, heterocyclic group, oxo, aryl and heteroaryl is optionally substituted at each occurrence by one or more substituents selected from deuterium, halogen, amino, nitro, cyano, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 6-10 aryl and a 5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O; Preferably, R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 2-4 cyano-alkyl, C 2-4 alkoxyalkyl, oxo group, -CONH2, C 3-6 alkenyl-C(O)NH-, C 2-4 alkynyl-NH-, -C(O)O-C 1-4 alkyl, -C(O)NH-C 1-4 alkyl, More preferably, R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 2-4 cyano-alkyl, C 2-4 alkoxy-C 1-4 alkyl, C 2-4 alkoxy-deuterated C 1-4 alkyl, oxo group, -CONH2, C 3-6 alkenyl-C(O)NH-, C 2-4 alkynyl-NH-, -C(O)O-C 1-4 alkyl, -C(O)NH-C 1-4 alkyl, 7. The compound according to claim 1, or its tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereoisomer or mixture thereof, or its deuterated derivative, or its pharmaceutically acceptable salt, solvate or prodrug, wherein, selected from 8. A compound according to any one of claims 1 to 7, or a tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein, the compound is a compound of formula (Ia)-(Ig): wherein, X1 is selected from CH, N, O or S; Y2 is selected from CH, CR y , N, O or S; M1 is selected from CH, N, O or S; M4 is selected from CH, N, O or S; R2 and R3 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group, -CONH2; Preferably, R2 and R3 are each independently selected from hydrogen, deuterium, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl; alternatively, R2 and R3 together with the C atom to which they are attached form a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S; Preferably, R2 and R3 together with the C atom to which they are attached form R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 2-6 cyano-alkyl, oxo, -CONH2, C 3-6 alkenyl-C(O)NH-, C 2-6 alkynyl-NH-, -C(O)O-C 1-6 alkyl, -C(O)NH-C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 alkyl-O-C 1-6 alkyl, -C 1-6 alkyl-O-C 3-6 cycloalkyl, C 1-6 alkyl-O-C 1-6 haloalkyl, deuterated C 1-6 alkyl-O-C 1-6 haloalkyl, C 1-6 alkyl-4- to 6-membered heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 1-6 alkyl-C 6-10 cycloalkyl, C 1-6 alkyl-C 6-10 aryl, C 1-6 alkyl-5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O, 4- to 6-membered saturated or partially unsaturated heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 6-10 aryl, 5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O, wherein each of said alkyl, haloalkyl, hydroxyalkyl, cycloalkyl, heterocyclic group, oxo, aryl, cycloalkyl, heterocyclic group and heteroaryl is optionally substituted, in each occurrence, by one or more substituents selected from halogen, amino, nitro, cyano, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 3-6 cycloalkyl, 4- to 6-membered heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 6-10 aryl and 5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O; Preferably, R5 is independently selected from i) hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 2-4 cyano-alkyl, oxo, -CONH2, C 3-6 alkenyl-C(O)NH-, C 2-4 alkynyl-NH-, -C(O)O-C 1-4 alkyl, -C(O)NH-C 1-4 alkyl, C 1-3 alkyl-O-C 1-5 alkyl, deuterated C 1-3 alkyl-O-C 1-3 haloalkyl, -C 1-3 alkyl-O-C 3-6 cycloalkyl, C 1-3 alkyl-O-C 1-6 haloalkyl, C 1-3 alkyl-O-N(C 1-3 alkyl)C(O)-, C 1-3 alkyl-N(C 1-3 alkyl)2, C 1-3 hydroxyalkoxy-C 1-3 alkyl, ii) optionally substituted by one or more substituents independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 2-4 cyano-alkyl, oxo, -CONH2, -C(O)O-C 1-4 alkyl, -C(O)NH-C 1-4 alkyl, C 1-3 alkyl-O-C 1-5 alkyl, deuterated C 1-3 alkyl-O-C 1-3 haloalkyl, C 1-3 alkyl-O-C 1-6 haloalkyl; R6 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl; Preferably, R6 is independently selected from hydrogen, deuterium, F, Cl, Br, amino, cyano, nitro, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy and C 1-6 hydroxyalkyl; R 2g 、R 3g 、R 4g 、R 5g each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, deuterated C 1-6 alkyl, deuterated C 2-6 alkenyl, deuterated C 2-6 alkynyl, deuterated C 1-6 haloalkyl, deuterated C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group, -CONH2; R y selected from hydrogen, deuterium, F, Cl, Br, amino, cyano, nitro, deuterated C 1-6 alkyl, deuterated C 2-6 alkenyl, deuterated C 2-6 alkynyl, deuterated C 1-6 haloalkyl, deuterated C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group and -CONH2; t is 0, 1, 2 or 3.

9. A compound of formula (II-a) or (II-b), or its tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereoisomer or mixture thereof, or its deuterated derivative, or its pharmaceutically acceptable salt, solvate or prodrug: wherein, L is NH, O, S, C 1-3 an alkylene group, -C 1-3 alkyl - O -, - C 1-3 alkyl - NH - or - CONH -; L1 is O, S or NH; Y2 is selected from CH, N, O or S; M4 is selected from CH, N, O or S; R2 and R3 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group and -CONH2; Alternatively, R2 and R3 together with the C atom to which they are attached form a C 3-6 cycloalkyl group or a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S; R4 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, oxo group, and -CONH2; R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1 or 2 heteroatoms selected from N or O, C 6-10 aryl, and a 5- to 7-membered heteroaryl containing 1 or 2 heteroatoms selected from N or O; Each R7 is independently selected from cyano, hydroxy, C 1-6 alkyl, deuterated C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, -NR a R b 、=NR b 、-C(O)NR a R b 、-NR a C(O)R b 、-C(O)OR b 、C 3-6 cycloalkyl, C 6-10 aryl, a 4- to 10-membered heterocyclic group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, a 5- to 10-membered heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S; wherein R7 is optionally substituted by one or more substituents selected from deuterium, halogen, amino, nitro, oxo, cyano, hydroxy, carboxy, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 1-6 alkoxy-C 1-6 alkyl, C 1-6 alkoxy-deuterated C 1-6 alkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S, a 5- to 6-membered heteroaryl containing 1 or 2 heteroatoms selected from N, O or S; Alternatively, R7 and together form a 9-10 membered heterocyclic group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, a 9-10 membered heteroaryl group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S; R a independently selected from hydrogen, deuterium, C 1-6 alkyl, deuterated C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 hydroxyalkyl and C 3-6 cycloalkyl; R b independently selected from hydrogen, deuterium, C 1-6 alkyl, deuterated C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S, and a 5- to 6-membered heteroaryl group containing 1 or 2 heteroatoms selected from N, O or S; m is 0, 1 or 2; n is 0, 1 or 2; t is 0, 1 or 2; and s is 0, 1, 2 or 3.

10. The compound according to claim 9, or its tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereoisomer or mixture thereof, or its deuterated derivative, or its pharmaceutically acceptable salt, solvate or prodrug, wherein, L is NH, O, S, L1 is O, S or NH; Y2 is selected from CH, N, O or S; M4 is selected from CH, N, O or S; R2 and R3 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy and C 1-3 hydroxyalkyl; Alternatively, R2 and R3, together with the C atom to which they are attached, form a C 3-6 cycloalkyl group or a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S; R4 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, oxo group and -CONH2; R5 is independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxy, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl and C 3-6 cycloalkyl; Each R7 is independently selected from cyano, hydroxy, C 1-3 alkyl, deuterated C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 hydroxyalkyl, -NR a R b 、=NR b 、-C(O)NR a R b 、-NR a C(O)R b 、-C(O)OR b 、C 3-6 cycloalkyl, phenyl, naphthyl, a 4- to 6-membered heterocyclic group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, a 5- to 6-membered heteroaryl group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S; wherein R7 is optionally substituted by one or more substituents selected from deuterium, halogen, amino, nitro, oxo, cyano, hydroxy, carboxyl, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 1-3 alkoxy-C 1-3 alkyl, C 1-3 alkoxy-deuterated C 1-3 alkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S and a 5- to 6-membered heteroaryl group containing 1 or 2 heteroatoms selected from N, O or S; R a independently selected from hydrogen, deuterium, C 1-3 alkyl, deuterated C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 hydroxyalkyl and C 3-6 cycloalkyl; R b independently selected from hydrogen, deuterium, C 1-3 alkyl, deuterated C 1-3 alkyl, C 1-3 alkoxy, C 1-3 haloalkyl, C 1-3 hydroxyalkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O or S, and a 5- to 6-membered heteroaryl group containing 1 or 2 heteroatoms selected from N, O or S.

11. A compound selected from the following: and the compounds of formula 1-12 Among them, Y2, L and M are each selected from the following groups, wherein, Y2, L and Z are each selected from the following groups, wherein, Y2 and L are each selected from the following groups, ； wherein, Y2 and L are each selected from the following groups, ； ； or a tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereoisomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

12. A method for preparing a compound of formula (I) or its pharmaceutically acceptable salt, which comprises the step of reacting a compound of formula (Xa) with a compound of formula (Xb), wherein, X is OH, SH or NH2; Hal is a halogen; R1, R2, R3, R4, R5, X1, X2, X3, X4, Y1, Y2, Y3, Y4, Y5, Y6, s and t are as defined in claim 1.

13. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to any one of claims 1-11, or its tautomer, cis- or trans-isomer, meso-form, racemate, enantiomer, diastereoisomer or mixture thereof, or its deuterated derivative, or its pharmaceutically acceptable salt, solvate or prodrug, and a pharmaceutically acceptable carrier.

14. A method for treating an MK2-regulated disorder or disease in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of claims 1-11, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a pharmaceutical composition according to claim 13.

15. A method for treating an immune disease, an autoimmune disease and an inflammatory disease, a cardiovascular disease, an infectious disease, a bone resorption disorder, a neurodegenerative disease in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of claims 1-11, or a tautomer, cis or trans isomer, meso form, racemate, enantiomer, diastereoisomer or mixture thereof, or a deuterated derivative thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a pharmaceutical composition according to claim 13.

16. The method according to claim 15, wherein the immune disease, autoimmune disease or inflammatory disease is selected from inflammatory bowel disease, ulcerative colitis, Crohn's disease, multiple sclerosis, psoriasis, arthritis, rheumatoid arthritis, osteoarthritis, juvenile arthritis, psoriatic arthritis, reactive arthritis, ankylosing spondylitis, cryopyrin-associated periodic syndromes, Muckle-Wells syndrome, familial cold autoinflammatory syndrome, neonatal-onset multisystem inflammatory disease, TNF receptor-associated periodic syndrome, acute and chronic pancreatitis, atherosclerosis, gout, ankylosing spondylitis, fibrotic diseases, liver fibrosis, idiopathic pulmonary fibrosis, kidney diseases, sarcoidosis, scleroderma, allergic reactions, diabetes, type 1 diabetes, type 2 diabetes, diabetic retinopathy, Still's disease, vasculitis, sarcoidosis, pulmonary inflammation, acute respiratory distress syndrome, wet and dry age-related macular degeneration, autoimmune hemolytic syndrome, autoimmune and inflammatory hepatitis, autoimmune neuropathy, autoimmune ovarian failure, autoimmune orchitis, autoimmune thrombocytopenia, silicone implant-associated autoimmune disease, Sjogren's syndrome, familial Mediterranean fever, systemic lupus erythematosus, vasculitis syndromes, temporal, Takayasu's and giant cell arteritis, Behcet's disease, Wegener's granulomatosis, vitiligo, secondary hematological manifestations of autoimmune diseases, anemia, drug-induced autoimmunity, Hashimoto's thyroiditis, hypophysitis, idiopathic thrombocytopenic purpura, metal-induced autoimmunity, myasthenia gravis, pemphigus, autoimmune deafness, Meniere's disease, Goodpasture's syndrome, Graves' disease, HW-associated autoimmune syndrome, Guillain-Barre syndrome, Addison's disease, antiphospholipid syndrome, asthma, atopic dermatitis, celiac disease, Cushing's syndrome, dermatomyositis, idiopathic adrenal cortical atrophy, idiopathic thrombocytopenia, Kawasaki syndrome, Lambert-Eaton syndrome, pernicious anemia, hay fever, polyarteritis nodosa, primary biliary cirrhosis, primary sclerosing cholangitis, Raynaud's syndrome, Reiter's syndrome, relapsing polychondritis, Schmidt's syndrome, thyroiditis, sepsis, septic shock, endotoxic shock, exotoxin-induced toxic shock, gram-negative sepsis, toxic shock syndrome, glomerulonephritis, peritonitis, interstitial cystitis, hyperoxia-induced inflammation, chronic obstructive pulmonary disease (COPD), vasculitis, graft-versus-host reaction, graft-versus-host disease, allograft rejection, acute allograft rejection, chronic allograft rejection, early graft rejection, acute allograft rejection, reperfusion injury, pain, acute pain, chronic pain, neuropathic pain, fibromyalgia, chronic infection, meningitis, encephalitis, myocarditis, gingivitis, post-surgical trauma, tissue injury, brain trauma, enterocolitis, sinusitis, uveitis, ocular inflammation, optic neuritis, gastric ulcer, esophagitis, peritonitis, periodontitis, dermatomyositis, gastritis, myositis, polymyalgia, pneumonia and bronchitis.

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