Alkynyl alcohol derivative and medical application thereof

Abstract

The invention discloses an alkynyl alcohol derivative and medical application thereof. The invention provides a compound shown as a formula (I) or pharmaceutically acceptable salt thereof. The invention further discloses a compound which is used for inhibiting NF-kappa B-induced kinase (NIK) and is shown in the formula (I), a synthetic route comprising the derivative and a corresponding intermediate thereof, a pharmaceutical composition of the compound and a use method of the pharmaceutical composition, and the compound and the pharmaceutical composition can be used for preparing medicines for preventing or treating NIK target mediated dependent diseases, such as NF-kappa B-induced kinase, further preventing or treating diseases such as cancers (such as b-cell malignancies including leukemia, lymphoma and myeloma), inflammatory diseases, autoimmune diseases and metabolic diseases such as obesity and diabetes mellitus. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and particularly relates to an alkynyl alcohol derivative and its medical use. Background Art

[0002] NF-κB plays a key regulatory role in cytokine-induced gene expression, participates in the body's inflammatory response and immune response, and can regulate cell apoptosis and stress response. Under physiological conditions, NF-κB exists in the cytoplasm in a low-activity state. Overactivation of NF-κB is associated with many human diseases such as inflammatory changes in rheumatoid arthritis, heart and brain diseases. The NF-κB transcription factor family has a total of five members: p105 / p50, p100 / p52, p65 (RelA), c-Rel, RelB, which are encoded by the NFκB1, NFκB2, RELA, REL, and RELB genes respectively. The members can form homologous or heterodimers to mediate signal transduction.

[0003] Depending on different activation mechanisms and functions, the NF-κB family is divided into the "canonical NF-κB pathway" and the "non-canonical NF-κB pathway; alternative NF-κB pathway". The canonical NF-κB pathway can mediate rapid and reversible inflammatory and immune responses; the non-canonical NF-κB pathway can mediate slower and irreversible physiological responses, and it has been found that it mainly regulates the differentiation and maturation of immune cells and the generation of secondary lymphoid organs. The non-canonical NF-κB signal responds to different signals from TNF superfamily receptors, including: CD40, CD27, lymphotoxin β receptor (LTβR), B cell activating factor receptor (BAFF-R), tumor necrosis factor receptor superfamily member 12A (TWEAK), etc. Under resting physiological conditions, TRAF3 participates in the connection of NIK to the TRAF2-cIAPs E3 complex, further promoting cIAP-mediated Lys48 ubiquitination and E3 ligase-mediated proteasomal degradation of NIK. Once the balance is disrupted, such as by the stimulation of cytokines CD40L or BAFF, NIK will accumulate in the cytoplasm, leading to phosphorylation of IKKα, processing of p100, and an increase in the protein level of p52. The generated p52 recruits RelB to form the p52 / RelB heterodimer, which translocates to the nucleus to trigger the expression of inflammation-related cytokines and chemokines. Known studies have shown that the normal transduction of the non-canonical NF-κB pathway regulates a variety of physiological processes, including: the differentiation, maturation, and survival of B cells, and the functions and immune behaviors of T cells. Similar to the canonical NF-κB signal, the NF-κB2 signal is also involved in specific aspects of the pathogenesis of RA. NIK is highly expressed in synovial endothelial cells of RA patients and promotes the formation of pathogenic blood vessels to exacerbate synovial inflammation by inducing chemokines (such as CXCL12) (Noort AR, van Zoest KP, Weijers EM, et al. NF-kappaB-inducing kinase is a key regulator of inflammation-induced and tumour-associated angiogenesis. The Journal of Pathology, 2014, 234(3):375-385.).Progressive joint damage in RA patients involves the abnormal formation and activation of osteoclasts, a process that depends on the NF-κB2 signaling pathway (Baum R, Gravallese EM. Bone as a target organ in rheumatic disease: impact on osteoclasts and osteoblasts. Clinical Reviews in Allergy&Immunology, 2016, 51(1): 1-15.). The pathogenesis of RA also involves the pathological function of B cells, mainly through the abnormal activation of the NF-κB2 pathway downstream of BAFF, to achieve an extended survival and maturation of B cells (Wei F, Chang Y, Wei Z. The role of BAFF in the progression of rheumatoid arthritis. Cytokine, 2015, 76(2): 537-544.). Similarly, SLE patients have higher serum BAFF levels, which are associated with the disease process. Among them, BAFF upregulates the activation degree of NF-κB2 signaling through NIK, promoting B cells to release autoantibodies to mediate the pathological survival and differentiation of B cells (Petri M, Stohl W, Chatham W, et al. Association of plasma B lymphocyte stimulator levels and disease activity in systemic lupus erythematosus. Arthritis Rheum, 2008, 58(8): 2453-2459.). At the same time, the overexpression of NIK leads to the dysregulated activation of NF-κB2, which plays an important role in mediating the pathogenesis of MS (Li Y, Wang H, Zhou X, et al. Cell intrinsic role of NF-κB-inducing kinase in regulating T cell-mediated immune and autoimmune responses. Scientific Reports, 2016, 6(1): 22115.). This is related to the abnormal activation of autoreactive Th1 and Th17 cells.NIK overexpression can promote the increased sensitivity of mice to Citrobacter rodentium in the intestine, thereby inducing intestinal inflammation (Simmons S, Pierson E, Lee S, et al. Modeling the heterogeneity of multiple sclerosis in animals. Trends in Immunology, 2013, 34(8): 410-422.).

[0004] The overactivation of NIK is related to the occurrence and development of tumors and autoimmune diseases, etc. However, there are few studies on the development and application of NIK inhibitors currently. Developing targeted small molecule NIK inhibitors in order to obtain candidate compounds with a brand-new mechanism of action for anti-tumor, autoimmune and inflammatory diseases and outstanding curative effects has gradually attracted the attention of many R & D institutions. Summary of the Invention

[0005] Object of the Invention: To solve the above technical problems, the present invention aims to provide a compound that can inhibit NF-κB-inducing kinase (NIK). The compound or its pharmaceutically acceptable salt can significantly inhibit NIK. As a NIK inhibitor, it can be used to prepare drugs for preventing or treating NIK target-mediated dependent diseases.

[0006] The present invention also provides a synthetic route for the derivatives and their corresponding intermediates of this kind and a pharmaceutical composition of the compound.

[0007] The present invention also provides the use of the compound or its pharmaceutically acceptable salt in the preparation of drugs for preventing or treating NIK target-dependent diseases.

[0008] Technical Solution: To achieve the above object, the present invention provides a compound as shown in the following formula (I) or its pharmaceutically acceptable salt:

[0009]

[0010] Wherein, represents a single bond or a double bond;

[0011] X 1 、X 4 、X 5 and X 6 each independently selected from C, N or CH;

[0012] X 2 is selected from N or CH;

[0013] X 3 is selected from O or CH;

[0014] R 1 is selected from hydrogen or C1 -C 6 alkyl;

[0015] R 2 selected from C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, 4- to 12-membered heterocyclic group, or 5- to 10-membered heteroaryl;

[0016] R 3 selected from hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 10 cycloalkyl, 4- to 7-membered heterocyclic group, 5- to 10-membered heteroaryl, or 5- to 10-membered aryl.

[0017] Furthermore, when represents a single bond, X 1 , X 2 and X 4 are N, X 3 is CH, X 5 and X 6 are each independently selected from N or CH.

[0018] Furthermore, when represents a double bond, X 1 is selected from C or N, X 2 and X 3 are selected from CH or O, X 4 is C, X 5 and X 6 are each independently selected from CH or N;

[0019] Furthermore, R 3 represents a 5-membered heteroaryl ring containing two or three heteroatoms each independently selected from O, S, and N; the 5-membered heteroaryl ring may optionally be substituted, when possible, on one ring N atom with a substituent selected from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 3 -C 10 cycloalkyl, 4- to 7-membered heterocyclic group, or C 1 -C 4 alkyl substituted with one, two, or three halogen atoms.

[0020] Furthermore, the R 3represents a 6-membered heteroaryl containing 1 or 2 N atoms, which is optionally substituted by one, two or three substituents, each of which is independently optionally selected from halogen, cyano, C 1 -C 6 alkyl, -O-C 1-4 alkyl, -O-C 3-6 cycloalkyl, C 3 -C 10 cycloalkyl, a 4- to 7-membered heterocyclic group or C 1 -C 4 alkyl substituted by one, two or three halogen atoms, or -O-C 1-4 alkyl substituted by one, two or three halogen atoms.

[0021] When R 3 represents an aryl group, it is optionally substituted by one, two or three substituents each independently optionally selected from the group consisting of halogen, cyano, C 1 -C 6 alkyl, -O-C 1-4 alkyl, -O-C 3-6 cycloalkyl, C 3 -C 10 cycloalkyl, a 4- to 7-membered heterocyclic group or C 1 -C 4 alkyl substituted by one, two or three halogen atoms, or -O-C 1-4 alkyl substituted by one, two or three halogen atoms.

[0022] Preferably, R 1 together with R 2 and the atoms to which it is attached forms a 4- to 6-membered heterocycloalkane ring or a 4- to 6-membered cycloalkane ring, and the 4- to 6-membered heterocycloalkane ring may be bicyclic.

[0023] Preferably, when R 3 is a 5-membered heteroaryl, it can be optionally selected from

[0024] Preferably, when R 3 is a 6-membered heteroaryl, it can be optionally selected from

[0025] More preferably, when R 3 is a 6-membered aryl, it can be optionally selected from

[0026] Among them, the compound is selected from any one of the following compounds:

[0027] 4-(4-(2-Aminopyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (1);

[0028] 4-(4-(2-(Methylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (2);

[0029] 4-(4-(2-(Isopropylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (3);

[0030] 4-(4-(2-(2-Methoxyethyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (4);

[0031] 4-(4-(2-(Cyclohexylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (5);

[0032] 4-(4-(2-(Bicyclo[1.1.1]pentan-1-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (6);

[0033] 4-(4-(2-(Pyridin-2-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (7);

[0034] 4-(4-(2-((4-Methoxypyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (8);

[0035] 4-(4-(2-((4,6-Dimethoxypyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (9);

[0036] 4-(4-(2-((6-Isopropoxypyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (10);

[0037] 2-((4-(2-(3-Hydroxy-3-(thiazol-2-yl)but-1-yn-1-yl)-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-yl)amino)isonicotinic acid (11);

[0038] 4-(4-(2-((4-(Dimethylamino)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (12);

[0039] 4-(4-(2-((6-Morpholinopyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (13);

[0040] 4-(4-(2-((6-((2-Methoxyethyl)(methyl)amino)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (14);

[0041] 4-(4-(2-((6-(Piperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (15);

[0042] 2-(Thiazol-2-yl)-4-(4-(2-((6-(Trifluoromethyl)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)but-3-yn-2-ol (16);

[0043] 4-(4-(2-((5-Fluoropyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (17);

[0044] 4-(4-(2-((4-Chloropyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (18);

[0045] 4-(4-(2-(Pyrimidin-4-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (19);

[0046] 4-(4-(2-(Quinolin-2-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (20);

[0047] 4-(4-(2-(Phenylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (21);

[0048] 4-(4-(2-((3-Methoxyphenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (22);

[0049] 3-((4-(2-(3-Hydroxy-3-(thiazol-2-yl)but-1-yn-1-yl)-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-yl)amino)benzonitrile (23);

[0050] 4-(4-(2-((3-(Dimethylamino)phenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (24);

[0051] 4-(4-(2-((3-Morpholinophenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (25);

[0052] 4-(4-(2-((3,5-Dimethoxyphenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (26);

[0053] 4-(4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (27);

[0054] 4-(4-(2-((2,2-Difluorobenzo[d][1,3]dioxol-5-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (28);

[0055] 4-(4-(2-((4-Fluorophenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (29);

[0056] 4-(4-(2-((4-Chlorophenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (30);

[0057] 4-(4-(2-((1-Methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (31);

[0058] 4-(4-(2-((1-Isopropyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (32);

[0059] 4-(4-(2-((1-Methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (33);

[0060] 4-(4-((4-Fluorophenyl)amino)-1,3,5-triazin-2-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (34);

[0061] 4-(4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-methylbut-3-yn-2-ol (35);

[0062] 3-((4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)ethynyl)oxetan-3-ol (36);

[0063] 1-((4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)ethynyl)cyclopent-1-ol (37);

[0064] 7-((4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)ethynyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-7-ol (38);

[0065] 4-(4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(5-methylisoxazol-3-yl)but-3-yn-2-ol (39);

[0066] 4-(7-(2-Aminopyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (40);

[0067] 4-(7-(2-(Bicyclo[1.1.1]pentan-1-ylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (41);

[0068] 4-(7-(2-((1-Methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (42);

[0069] 4-(7-(2-((1-Methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (43);

[0070] 4-(7-(2-((1-(Tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (44);

[0071] 4-(7-(2-((1-(2-(Dimethylamino)ethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (45);

[0072] 4-(7-(2-((1-Methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (46);

[0073] 4-(7-(2-(phenylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (47);

[0074] 4-(7-(2-((3-Methoxyphenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (48);

[0075] 3-((4-(2-(3-Hydroxy-3-(thiazol-2-yl)but-1-yn-1-yl)benzofuran-7-yl)pyrimidin-2-yl)amino)benzonitrile (49);

[0076] 4-(7-(2-((3-(Dimethylamino)phenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (50);

[0077] 4-(7-(2-((3-Morpholinophenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (51);

[0078] 4-(7-(2-((4-Fluorophenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (52);

[0079] 4-(7-(2-(((4-Chlorophenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (53);

[0080] 4-(7-(2-(benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (54);

[0081] 4-(7-(2-((2,3-Dihydrobenzo[b][1,4]dioxan-6-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (55);

[0082] 4-(7-(2-(pyridin-2-ylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (56);

[0083] 4-(7-(2-((6-(Dimethylamino)pyridin-2-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (57);

[0084] 4-(7-(2-((6-Morpholinopyridin-2-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (58);

[0085] 4-(7-(2-((6-Methoxypyridin-2-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (59);

[0086] 4-(7-(2-((2-(Difluoromethyl)pyridin-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (60).

[0087]

[0088]

[0089]

[0090]

[0091] Furthermore, the pharmaceutically acceptable salts include acid addition salts formed by the compound of general formula (I) and the following acids: the acids are hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, or acid salts of inorganic bases containing alkaline metal cations, alkaline earth metal cations or ammonium cations.

[0092] Use of the compound or its pharmaceutically acceptable salt according to the present invention in the preparation of a NIK inhibitor.

[0093] Use of the compound or its pharmaceutically acceptable salt according to the present invention in the preparation of a drug for preventing or treating NIK target-mediated dependent diseases.

[0094] Furthermore, the NIK target-mediated related diseases are tumor diseases and autoimmune diseases.

[0095] Further, the NIK target-mediated dependent diseases are rheumatoid arthritis, gouty arthritis, systemic lupus erythematosus, Crohn's disease, ulcerative colitis, inflammatory bowel disease, psoriatic arthritis, psoriasis, type I diabetes, atopic dermatitis, myelofibrosis hyperplasia, polycythemia vera, acute hepatorenal toxic injury, myeloma, sepsis or acute lymphocytic leukemia.

[0096] Wherein, the present invention provides a pharmaceutical composition comprising the compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof, or one or more pharmaceutically acceptable carriers or excipients.

[0097] Further, the use of the pharmaceutical composition in the preparation of a medicament for preventing or treating NIK-mediated diseases.

[0098] Further, the pharmaceutical composition is a capsule, powder, tablet, granule, pill, injection, syrup, oral preparation, inhalant, ointment, suppository or patch.

[0099] The term "optionally" or "optionally" means that the subsequent described event or situation may or may not occur, and this description includes the occurrence and non-occurrence of the described event or situation. For example, ethyl is "optionally" substituted by one or more halogens, which means that ethyl can be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl, etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2, etc.) or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3, etc.). Those skilled in the art will understand that for any group containing one or more substituents, no substitution or substitution pattern that is spatially impossible to exist and / or cannot be synthesized will be introduced.

[0100] The term "alkyl" refers to a hydrocarbon group of the general formula C n H 2n+1 and the alkyl can be straight-chain or branched-chain. The term "C 1 -C 10"Alkyl" can be understood to represent a straight-chain or branched-chain saturated hydrocarbon group having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Specific examples of the alkyl group include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, etc.; the term "C 1 -C 6 alkyl" can be understood to represent an alkyl group having 1 to 6 carbon atoms. Specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C 1 -C 3 alkyl" can be understood to represent a straight-chain or branched-chain saturated alkyl group having 1 to 3 carbon atoms. The "C 1 -C 10 alkyl" can include ranges such as "C 1 -C 6 alkyl" or "C 1 -C 3 alkyl", etc. The "C 1 -C 6 alkyl" can further include "C 1 -C 3 alkyl".

[0101] The term "alkoxy" refers to a group formed by removing the hydrogen atom on the hydroxyl group of a straight-chain or branched-chain alcohol, and can be understood as "alkyloxy" or "alkyl-O-". The term "C 1 -C 6 alkoxy" can be understood as "C 1 -C 6 alkyloxy" or "C 1 -C 6 alkyl-O-". The "C 1 -C 6 alkoxy" can further include "C 1 -C 3 alkoxy".

[0102] The term "cycloalkyl" or "cycloalkane" refers to a carbocyclic group that is completely saturated and exists in the form of a monocyclic, fused-ring, bridged-ring, or spiro-ring, etc. Unless otherwise indicated, the carbocyclic ring is usually a 3- to 20-membered ring. The term "C 3-C 10 "Cycloalkyl" refers to cycloalkyl groups having 3, 4, 5, 6, 7, 8, 9 or 10 ring carbon atoms. The term "C 3 -C 6 ycloalkyl" refers to cycloalkyl groups having 3, 4, 5 or 6 ring carbon atoms.

[0103] The term "heterocyclic group" refers to a fully saturated or partially saturated (not aromatic heterocyclic as a whole) monocyclic, fused-ring, spiro or bridged-ring group, which contains 1-5 (e.g., 1-3 or 1-2) heteroatoms or heteroatom groups (i.e., atomic groups containing heteroatoms) among its ring atoms. The "heteroatoms or heteroatom groups" include, but are not limited to, nitrogen atom (N), oxygen atom (O), sulfur atom (S), phosphorus atom (P), boron atom (B), -S(=O) 2 -, -S(=O)-, -P(=O) 2-, -P(=O)-, -NH-, -S(=O)(=NH)-, -C(=O)NH- or -NHC(=O)NH-, etc. The term "4- to 12-membered heterocyclic group" refers to a heterocyclic group having 4, 5, 6, 7, 8, 9, 10, 11 or 12 ring atoms, and having 1 to 5 heteroatoms or heteroatom groups independently selected from the above-mentioned ones in its ring atoms. The "4- to 12-membered heterocyclic group" may include the "4- to 7-membered heterocyclic group". The term "4- to 7-membered heterocyclic group" refers to a heterocyclic group having 4, 5, 6 or 7 ring atoms, and having 1, 2, 3, 4 or 5 heteroatoms or heteroatom groups independently selected from the above-mentioned ones in its ring atoms. Among them, specific examples of the 4-membered heterocyclic group include, but are not limited to, azetidinyl or oxetanyl; specific examples of the 5-membered heterocyclic group include, but are not limited to, tetrahydrofuranyl, dioxolanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrroline, 4,5-dihydrooxazolyl or 2,5-dihydro-1H-pyrrolyl; specific examples of the 6-membered heterocyclic group include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridyl or 4H-[1,3,4]thiadiazinyl; specific examples of the 7-membered heterocyclic group include, but are not limited to, diazepanyl. The heterocyclic group may also be a bicyclic group. Among them, specific examples of the 5,5-bicyclic group include, but are not limited to, hexahydrocyclopenta[c]pyrrol-2(1H)-yl; specific examples of the 5,6-bicyclic group include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group may be a benzo-fused ring group of the above-mentioned 4- to 7-membered heterocyclic group, and specific examples include, but are not limited to, dihydroisoquinolinyl, etc. The "4- to 12-membered heterocyclic group" may include ranges such as "7- to 12-membered heterocyclic group", "6- to 11-membered heterocyclic group", "5- to 10-membered heterocyclic group", "4- to 7-membered heterocyclic group", "5- to 6-membered heterocyclic group", "6- to 8-membered heterocyclic group", "4- to 10-membered heterocycloalkyl group", "5- to 10-membered heterocycloalkyl group", "4- to 7-membered heterocycloalkyl group", "5- to 6-membered heterocycloalkyl group", "6- to 8-membered heterocycloalkyl group", etc. The "4- to 7-membered heterocyclic group" may further include ranges such as "4- to 6-membered heterocyclic group", "5- to 6-membered heterocyclic group", "4- to 7-membered heterocycloalkyl group", "4- to 6-membered heterocycloalkyl group", "5- to 6-membered heterocycloalkyl group", etc. Although some bicyclic heterocyclic groups in the present disclosure partially contain a benzene ring or a heteroaryl ring, the heterocyclic group as a whole is still non-aromatic.

[0104] The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π-electron system. The aryl may have 6 to 20 carbon atoms, 6 to 14 carbon atoms or 6 to 12 carbon atoms. The term "C 6 -C 10"Aryl" can be understood as an aryl group having 6 to 10 carbon atoms. For example, a ring having 6 carbon atoms ("C 6 aryl"), such as a phenyl group; or a ring having 9 carbon atoms ("C 9 aryl"), such as an indanyl group or an indenyl group; or a ring having 10 carbon atoms ("C 10 aryl"), such as a tetrahydronaphthyl group, a dihydronaphthyl group or a naphthyl group.

[0105] The term "heteroaryl" refers to a monocyclic or fused polycyclic system having aromaticity, wherein the ring atoms contain at least one ring atom selected from N, O, S, and the remaining ring atoms are aromatic ring groups of C. The term "5-10 membered heteroaryl" can be understood to include such monocyclic or bicyclic aromatic ring systems: which have 5, 6, 7, 8, 9 or 10 ring atoms, such as 5 or 6 or 9 or 10 ring atoms, and which contain 1-5, such as 1-3 heteroatoms independently selected from N, O and S. In particular, heteroaryl is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl, etc. and their benzo derivatives, such as benzofuryl, benzothienyl, benzothiazolyl, benzoxazolyl, benzoisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl, etc. and their benzo derivatives, such as quinolinyl, quinazolinyl or isoquinolinyl, etc.; or azocinyl, indolizinyl, purinyl, etc. and their benzo derivatives; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl or phenoxazinyl, etc. The term "5-membered heteroaryl" refers to an aromatic ring system having 5 ring atoms, and which contains 1-3, such as 1-2 heteroatoms independently selected from N, O and S.

[0106] Advantages: Compared with the prior art, the present invention has the following remarkable advantages:

[0107] Compared with the prior art, the present invention provides a design and preparation method for a class of compounds with novel structural features and further explores the application of the compounds. By analyzing the in vitro activity data at the molecular level, it is shown that: the compounds of the present invention have significant protein inhibitory activity against NIK, and the medicinal combination types of these compounds and the potential application value of this class of compounds as NIK inhibitors in the prevention or treatment of inflammatory diseases and tumors and other directions are confirmed. BRIEF DESCRIPTION OF THE DRAWINGS

[0108] Figure 1 It is the inhibitory effect of compound 27 on the secretion of macrophage inflammatory factors. DETAILED DESCRIPTION OF THE INVENTION

[0109] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0110] The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified. The experimental methods without specific conditions in the examples are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer.

[0111] Example 1

[0112] The present invention synthesizes the compound of general formula (I) or its pharmaceutically acceptable salt by the following method: The synthesis method of the above general formula (I) includes the following steps:

[0113] Method I-1

[0114]

[0115] Method I-2

[0116]

[0117] The reaction conditions are as follows: a) glacial acetic acid, 110 °C, 0.5 h; b) sodium borohydride, methanol, RT, 3 h; c) p-toluenesulfonic acid hydrate, tert-butanol, 90 °C, 12 h; d) sodium iodide, copper iodide, N,N'-dimethylethylenediamine, anhydrous dioxane, N 2 , 110 °C, 24 h; e) copper iodide, bis(triphenylphosphine)palladium dichloride, triethylamine / tetrahydrofuran (1 / 3), N 2 , 40 °C, 3 h; f) tris(dibenzylideneacetone)dipalladium, 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, cesium carbonate, anhydrous dioxane, N 2 , 90 °C, 4 h; g) sodium hydride, N,N-dimethylformamide, 0 °C - rt, 3 h; h) neopentyl glycol diborate, DPPF palladium dichloride, potassium acetate, dioxane, N 2 , 90 °C, 12 h; i) DPPF palladium dichloride, potassium carbonate, dioxane / water (5 / 1), N 2 , 90 °C, 3 h; j) iodine, lithium diisopropylamide, THF, N 2 , -78 °C, 4 h; k) methanol solution of ammonia, 90 °C, 12 h.

[0118] The compounds of the present invention can all be prepared by the above or similar preparation methods, and the corresponding raw materials can be selected according to the different substituents and substitution positions.

[0119] Example 2

[0120] 2-Bromopyrazolo[1,5-a]pyrimidine (I-a)

[0121] Weigh 10.0 g (61.7 mmol) of 3-bromo-1H-pyrazol-5-amine and 6.1 g (61.7 mmol) of 3-dimethylaminopropenal and add them to a 250 ml reaction flask. Then add 100 ml of glacial acetic acid. Stir the reaction at 110 °C for 0.5 h. After the reaction is complete, concentrate the reaction solution under vacuum and perform column chromatography (PE / EA = 5:1). Obtain 11.2 g of I-a as a yellow solid, with a yield of 91%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.11 (ddd, J = 7.0, 1.8, 0.9 Hz, 1H), 8.60 (dd, J = 4.1, 1.7 Hz, 1H), 7.10 (dd, J = 7.1, 4.1 Hz, 1H), 6.94 (d, J = 1.0 Hz, 1H). MS (m / z): [M+H] + 198.0.

[0122] Example 3

[0123] 2-Bromo-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine (I-b)

[0124] Weigh 2.0 g (10.1 mmol) of I-a prepared in Example 2 and add it to a 100 ml reaction flask. Then add 50 ml of methanol. Slowly add 1.53 g (40.4 mmol) of sodium borohydride to the above reaction solution under vigorous stirring and react at room temperature for 3 h. After the reaction is complete, concentrate the reaction solution under vacuum and perform column chromatography (PE / EA = 3:1). Obtain 2.0 g of I-b as a white solid, with a yield of 98%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 6.27 (s, 1H), 5.26 (s, 1H), 3.92 (t, J = 6.1 Hz, 2H), 3.21–

[0125] 3.07 (m, 2H), 1.97 (p, J = 6.1 Hz, 2H). MS (m / z): [M+H] + 202.0.

[0126] Example 4

[0127] 4-(2-Bromo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-amine (I-c)

[0128] Weigh 1 g (5 mmol) of Ⅰ-b prepared in Example 3, 650 mg (5 mmol) of 2-amino-4-chloropyrimidine, and 1.9 g (10.0 mmol) of p-toluenesulfonic acid hydrate into a high-temperature pressure-resistant reaction tube, and add 20 ml of tert-butanol. Stir the reaction at 90 °C for 12 hours. After the reaction is complete, add 50 ml of saturated sodium bicarbonate aqueous solution, and extract the resulting mixture with ethyl acetate (50 ml × 3). Combine the organic layers and wash with saturated brine, Na 2 S0 4 Dry, concentrate in vacuo, and perform column chromatography (PE / EA = 1:1) to obtain 1.1 g of Ⅰ-c as a pale yellow solid, with a yield of 75.3%. This preparation method is a general preparation method, and this method is also applicable to the synthesis of intermediates Ⅰ-f and Ⅰ-k. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.02 (d, J = 6.0 Hz, 1H), 6.97 (s, 1H), 6.54 (s, 2H), 6.31 (d, J = 6.0 Hz, 1H), 4.10 (t, J = 6.1 Hz, 2H), 3.79 (t, J = 5.8 Hz, 2H), 2.21–2.10 (m, 2H). [M+H] + 295.0.

[0129] Example 5

[0130] 4-(2-Iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-amine (Ⅰ-d)

[0131] Weigh 1.2 g (4.1 mmol) of Ⅰ-c prepared in Example 4, 1.2 g (8.2 mmol) of sodium iodide, and 456 mg (2.4 mmol) of copper(I) iodide into a 50 ml reaction flask, and add 20 ml of anhydrous dioxane. After purging with nitrogen, add 361 mg (4.1 mmol) of N,N'-dimethylethylenediamine. Stir the reaction at 110 °C for 24 hours. After the reaction is complete, add 50 ml of pure water, and extract the resulting mixture with ethyl acetate (50 ml × 3). Combine the organic layers and wash with saturated brine, Na 2 S0 4 Dry, concentrate in vacuo. The obtained crude product Ⅰ-d can be directly applied to the next reaction without purification.

[0132] Example 6

[0133] 4-(4-(2-Aminopyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 1)

[0134] Weigh 130 mg (0.38 mmol) of Compound I-d prepared in Example 5, 116 mg (0.76 mmol) of 2-(thiazol-2-yl)but-3-yn-2-ol, 13.3 mg (0.02 mmol) of bis(triphenylphosphine)palladium(II) dichloride, 7.3 mg (0.04 mmol) of copper(I) iodide into a 25 ml reaction flask. Add a mixed solvent of tetrahydrofuran and triethylamine 4.5 / 1.5 ml (THF / Et 3 N = 3:1) to the reaction mixture. Evacuate and backfill with N 2 Place the reaction under stirring at 40 °C for 2.5 h. After the reaction is complete, concentrate the reaction solution, and perform column chromatography (PE:EA = 1:2) to obtain 95 mg of Compound 1 as a yellow solid, with a yield of 67.9%. This preparation method is a general preparation method, and this method is also applicable to the synthesis of Compounds 2-6 and 21-55. 1 1H NMR (300 MHz, DMSO-d 6 ) δ 8.00 (d, J = 6.0 Hz, 1H), 7.77 (d, J = 3.2 Hz, 1H), 7.69 (d, J = 3.2 Hz, 1H), 7.06 (s, 1H), 6.95 (s, 1H), 6.56 (s, 2H), 6.32 (d, J = 6.0 Hz, 1H), 4.12 (t, J = 6.0 Hz, 2H), 3.81 (t, J = 6.0 Hz, 2H), 2.23 - 2.09 (m, 2H), 1.86 (s, 3H). [M+H] + 368.1.

[0135] Example 7

[0136] 4-(4-(2-(Pyridin-2-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 7)

[0137] Weigh 200 mg (0.54 mmol) of Compound 1 prepared in Example 6, 103 mg (0.65 mmol) of 2-bromopyridine, 24 mg (0.03 mmol) of tris(dibenzylideneacetone)dipalladium(0), 31 mg (0.05 mmol) of 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, and 264 mg (0.81 mmol) of cesium carbonate into a 25 ml reaction flask. Add 10 ml of anhydrous dioxane, evacuate and backfill with N 2 Place the reaction under stirring at 90 °C for 3 h. After the reaction is complete, concentrate the reaction solution, and perform column chromatography (EA) to obtain 150 mg of Compound 7 as a yellow solid, with a yield of 62.5%. This preparation method is a general preparation method, and this method is also applicable to the synthesis of Compounds 8-20 and 56-60. 11H NMR (300 MHz, DMSO-d 6 ) δ 9.91 (s, 1H), 8.29 (t, J = 5.3 Hz, 2H), 8.10 (d, J = 8.4 Hz, 1H), 7.79 (d, J = 3.3 Hz, 1H), 7.76–7.63 (m, 2H), 7.30 (s, 1H), 7.07 (s, 1H), 6.94 (dd, J = 7.4, 5.0 Hz, 1H), 6.67 (d, J = 6.1 Hz, 1H), 4.15 (t, J = 6.0 Hz, 2H), 3.89 (t, J = 5.9 Hz, 2H), 2.26 - 2.13 (m, 2H), 1.89 (s, 3H). MS (m / z): [M+H] + 445.1.

[0138] Example 8

[0139] 2-Bromo-4-(2-chloropyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine (Ⅰ-e)

[0140] Weigh 202 mg (1 mmol) of Ⅰ-b prepared in Example 3 into a 25 ml reaction flask, add 10 ml of N,N-dimethylformamide, and slowly add 48 mg (2 mmol) of sodium hydride under ice bath stirring. After addition, continue stirring under ice bath for 30 minutes. Weigh 298 mg (2 mmol) of 2,4-dichloropyrimidine and add it to the above reaction solution. Transfer to room temperature and continue the reaction for 3 hours. After the reaction is complete, add 50 ml of water, and the resulting mixture is extracted with ethyl acetate (50 ml × 3). Combine the organic layers and wash with saturated brine, Na 2 SO 4 dry, and concentrate under vacuum. Column chromatography (PE / EA = 1:2) gives 280 mg of compound Ⅰ-e as a yellow solid, with a yield of 89.2%. 1 1H NMR (300 MHz, DMSO-d6) δ 8.55 (d, J = 6.1 Hz, 1H), 7.36 (d, J = 6.1 Hz, 1H), 6.88 (s, 1H), 4.28 (t, J = 6.0 Hz, 2H), 4.05 (t, J = 5.9 Hz, 2H), 2.40 - 2.28 (m, 2H). MS (m / z): [M+H] + : 313.6.

[0141] Example 9

[0142] Using the preparation method of Example 4, with the only difference being that compound I-b is replaced by compound I-e prepared in Example 8, and 2-amino-4-chloropyrimidine is replaced by a primary amine derivative, and other processes remain unchanged, compound I-f is prepared (including compound I-f-1, when synthesizing I-f-1, the primary amine derivative is aniline, i.e., II-1 in the synthesis route).

[0143] Example 10

[0144] Using the preparation method of Example 5, with the only difference being that compound I-c is replaced by compound I-f prepared in Example 9, and other processes remain unchanged, compound I-g is prepared (including compounds I-g-1 and I-h).

[0145] Example 11

[0146] 2-(Benzofuran-7-yl)-5,5-dimethyl-1,3,2-dioxaborolane (I-i)

[0147] Weigh 1 g (5.08 mmol) of 7-bromobenzofuran, 1.6 g (7.10 mmol) of neopentyl glycol diborate, 260 mg (0.36 mmol) of DPPF palladium dichloride, and 1.5 g (15.2 mmol) of potassium acetate into a 50 ml reaction flask, add 20 ml of dioxane, and evacuate and replace with N 2 Place the reaction in a stirrer at 80 °C for 12 hours. After the reaction is complete, filter by suction and concentrate the reaction solution. The crude product I-i can be directly applied to the next reaction without purification.

[0148] Example 12

[0149] 4-(Benzofuran-7-yl)-2-chloropyrimidine (I-j)

[0150] Weigh 1 g (4.3 mmol) of compound I-i prepared in Example 11, 777 mg (5.2 mmol) of 2,4-dichloropyrimidine, 160 mg (0.21 mmol) of DPPF palladium dichloride, and 1.28 g (13.0 mmol) of potassium acetate into a 50 ml reaction flask, 15 ml / 3 ml of a mixed solvent of dioxane and water (dioxane / H 2 O = 5:1), evacuate and replace with N 2 Place the reaction in a stirrer at 80 °C for 12 hours. After the reaction is complete, filter by suction and concentrate the reaction solution. Column chromatography (PE / EA = 1:5) gives 830 mg of compound I-j as a yellow solid, with a yield of 82.7%. 1 H NMR (300 MHz, DMSO-d 6)δ 8.90 (d, J = 5.3 Hz, 1H), 8.35 (d, J = 5.4 Hz, 1H), 8.18 (d, J = 11.8 Hz, 2H), 7.92 (d, J = 7.8 Hz, 1H), 7.45 (t, J = 7.8 Hz, 1H), 7.14 (s, 1H). MS (m / z): [M+H] + 231.0.

[0151] Example 13

[0152] Using the preparation method of Example 4, the difference is only that compound I-b is replaced with compound I-j prepared in Example 12, and 2-amino-4-chloropyrimidine is replaced with a primary amine derivative, and other processes remain unchanged, to prepare compound I-k (including compound I-k-1. When synthesizing I-k-1, the primary amine derivative is aniline, that is, II-2 in the synthesis route).

[0153] Example 14

[0154] 4-(2-Iodobenzofuran-7-yl)-N-phenylpyrimidin-2-amine (I-l-1)

[0155] Weigh 287 mg (1 mmol) of I-k-1 prepared in Example 13 into a 25 ml reaction flask, evacuate and replace with N 2 , add 10 ml of anhydrous tetrahydrofuran, place the reaction at -78 °C, slowly add 1.5 ml (3 mmol, 2 mol / L) of LDA, and continue to stir at -78 °C for 0.5 hour. Weigh 762 mg (3 mmol) of iodine and dissolve it in 2 ml of anhydrous tetrahydrofuran, slowly add it to the above reaction solution, and continue to stir at -78 °C for 3 hours. After the reaction is complete, transfer it to room temperature, and successively add saturated NH 4 Cl and saturated Na 2 S 2 O 3 solution to quench. The resulting mixture is extracted with ethyl acetate (50 ml × 3). Combine the organic layers, wash with saturated brine, and dry over Na 2 S0 4 , and concentrate under vacuum. The obtained crude product I-l-1 can be directly applied to the next reaction without purification. This preparation method is a general preparation method and can be used for the synthesis of compound I-l from compound I-k.

[0156] Example 15

[0157] 4-(Benzofuran-7-yl)pyrimidin-2-amine (I-m)

[0158] Weigh 1 g (4.35 mmol) of compound I-j prepared in Example 12 into a high-temperature pressure-resistant reaction tube, add ammonia (NH 3) 20 mL (2.5 mmol / L) of a methanol solution, and the reaction was stirred at 90 °C for 12 hours. After the reaction was complete, it was concentrated under vacuum, and column chromatography (PE / EA = 1:1) gave 0.82 g of I-m as a pale yellow solid, with a yield of 89.1%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.41 (d, J = 5.1 Hz, 1H), 8.12 (dd, J = 8.1, 1.8 Hz, 2H), 7.81 (dd, J = 7.7, 1.4 Hz, 1H), 7.48 (d, J = 5.1 Hz, 1H), 7.40 (t, J = 7.7 Hz, 1H), 7.09 (d, J = 2.3 Hz, 1H), 6.75 (s, 2H). MS (m / z): [M+H] + 212.1.

[0159] Example 16

[0160] Using the preparation method of Example 14, the only difference was that compound I-k was replaced with compound I-m prepared in Example 15, and the other procedures remained unchanged to obtain compound I-n.

[0161] Example 17

[0162] 4-(4-(2-(Methylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 2)

[0163] Using the preparation method of Example 6, replacing I-d with 4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)-N-methylpyrimidin-2-amine, and keeping the other procedures unchanged, 120 mg of Compound 2 (white solid) was obtained, with a yield of 84.2%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.05 (d, J = 5.8 Hz, 1H), 7.78 (d, J = 3.2 Hz, 1H), 7.69 (d, J = 3.3 Hz, 1H), 7.06 (s, 1H), 7.04–6.95 (p, J = 4.16 Hz, 1H), 6.89 (s, 1H), 6.32 (d, J = 5.9 Hz, 1H), 4.12 (t, J = 6.0 Hz, 2H), 3.83 (t, J = 4.5 Hz, 2H), 2.80 (d, J = 4.7 Hz, 3H), 2.24-2.08 (m, 2H H), 1.86 (s, 3H). MS (m / z): [M+H] + 382.1.

[0164] Example 18

[0165] 4-(4-(2-(Isopropylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 3)

[0166] Using the preparation method of Example 6, replace I-d with 4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)-N-isopropylpyrimidin-2-amine, and keep other processes unchanged, to obtain 100 mg of Compound 3 (white solid) with a yield of 80.5%. 1 H NMR(300MHz,DMSO-d 6 ) δ8.04(d,J = 5.9Hz,1H),7.78(d,J = 3.2Hz,1H),7.70(d,J = 3.2Hz,1H),7.06(s,1H),7.02 - 6.70(m,2H),6.30(d,J = 5.9Hz,1H),4.12(t,J = 6.0Hz,2H),4.08 - 3.93(m,1H),3.82(t,J = 5.7Hz,2H),2.23 - 2.08(m,2H),1.87(s,3H),1.15(d,J = 6.5Hz,6H).MS(m / z):[M+H] + 410.1.

[0167] Example 17

[0168] 4-(4-(2-(2-Methoxyethyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 4)

[0169] Using the preparation method of Example 6, replace I-d with 4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)-N-(2-methoxyethyl)pyrimidin-2-amine, and keep other processes unchanged, to obtain 90 mg of Compound 4 (white solid) with a yield of 79.8%. 1 H NMR(300MHz,DMSO-d 6)δ 8.05 (d, J = 5.6 Hz, 1H), 7.77 (d, J = 3.2 Hz, 1H), 7.69 (d, J = 3.2 Hz, 1H), 7.32 - 6.58 (m, 3H), 6.33 (d, J = 5.4 Hz, 1H), 4.12 (t, J = 6.0 Hz, 2H), 3.83 (t, J = 5.6 Hz, 2H), 3.49 - 3.38 (m, 4H), 3.25 (s, 3H), 2.24 - 2.05 (m, 2H), 1.86 (s, 3H). MS (m / z): [M + H] + 426.2。

[0170] Example 17

[0171] 4-(4-(2-(Cyclohexylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 5)

[0172] Using the preparation method of Example 6, replace Ⅰ-d with N-cyclohexyl-4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-amine, and keep other processes unchanged, to obtain 133 mg of Compound 5 (white solid), with a yield of 88.1%. 1 H NMR (300 MHz, DMSO-d 6 )δ 8.03 (d, J = 5.9 Hz, 1H), 7.76 (d, J = 3.3 Hz, 1H), 7.69 (d, J = 3.2 Hz, 1H), 7.04 (s, 1H), 6.94 (d, J = 8.0 Hz, 2H), 6.25 (d, J = 5.9 Hz, 1H), 4.12 (t, J = 5.9 Hz, 2H), 3.79 (t, J = 5.8 Hz, 2H), 3.74 - 3.50 (m, 1H), 2.21 - 2.10 (m, 2H), 1.97 - 1.82 (m, 5H), 1.76–1.66 (m, 2H), 1.64–1.53 (m, 1H), 1.35–1.11 (m, 5H). MS (m / z): [M + H] + 450.2.

[0173] Example 18

[0174] 4-(4-(2-(Bicyclo[1.1.1]pent-1-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 6)

[0175] Using the preparation method of Example 6, replace Ⅰ-d with N-(bicyclo[1.1.1]pentan-1-yl)-4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-amine, and keep other processes unchanged to obtain Compound 6 (white solid).

[0176] 146 mg, yield 87.3%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.06 (s, 1H), 7.87 - 7.72 (m, 2H), 7.69 (d, J = 3.2 Hz, 1H), 7.06 (s, 1H), 6.77 (s, 1H), 6.42 (d, J = 5.3 Hz, 1H), 4.13 (t, J = 6.0 Hz, 2H), 3.87 (t, J = 4.89 Hz, 2H), 2.42 (s, 1H), 2.24 - 2.11 (m, 2H), 2.06 (s, 6H), 1.87 (s, 3H). MS (m / z): [M+H] + 434.2.

[0177] Example 19

[0178] 4-(4-(2-((4-Methoxypyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 8)

[0179] Using the preparation method of Example 7, replace 2-bromopyridine with 2-bromo-4-methoxypyridine, and keep other processes unchanged to obtain Compound 8 (white solid) 132 mg, yield 84.3%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.84 (s, 1H), 8.29 (d, J = 5.9 Hz, 1H), 8.11 (d, J = 5.8 Hz, 1H), 7.83 – 7.75 (m, 2H), 7.70 (d, J = 3.2 Hz, 1H), 7.25 (s, 1H), 7.06 (s, 1H), 6.69 (d, J = 6.0 Hz, 1H), 6.57 (dd, J = 5.8, 2.3 Hz, 1H), 4.15 (t, J = 6.0 Hz, 2H), 3.92 (t, J = 5.3 Hz, 2H), 3.81 (s, 3H), 2.26 - 2.14 (m, 2H), 1.88 (s, 3H). MS (m / z): [M+H] + 475.2.

[0180] Example 20

[0181] 4-(4-(2-((4,6-Dimethoxypyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 9)

[0182] Using the preparation method of Example 7, replace 2-bromopyridine with 2-bromo-4,6-dimethoxypyridine, and keep other processes unchanged. 101 mg of Compound 9 (white solid) was obtained with a yield of 75.3%. 1 H NMR(300MHz,DMSO-d 6 )δ9.54(s,1H),8.28(d,J=5.8Hz,1H),7.77(d,J=3.2Hz,1H),7.68(d,J=3.3Hz,1H),7.55(d,J=1.9Hz,1H),7.14(s,1H),7.04(s,1H),6.69(d,J=5.8Hz,1H),5.96(d,J=1.9Hz,1H),4.15(t,J=5.9Hz,2H),3.92(t,J=5.7Hz,2H),3.84(s,3H),3.77(s,3H),2.25-2.14(m,2H),1.87(s,3H).MS(m / z):[M+H] + 505.2.

[0183] Example 21

[0184] 4-(4-(2-((6-Isopropoxypyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 10)

[0185] Using the preparation method of Example 7, replace 2-bromopyridine with 2-bromo-6-isopropoxypyridine, and keep other processes unchanged. 123 mg of Compound 10 (white solid) was obtained with a yield of 85.6%. 1 H NMR(400MHz,DMSO-d 6)δ9.52(s,1H),8.28(d,J=6.0Hz,1H),7.81–7.73(m,2H),7.70(d,J=3.3Hz,1H),7.57(t,J=8.0Hz,1H),7.08(d,J=15.0Hz,2H),6.68(d,J=6.0Hz,1H),6.29(d,J=8.0Hz,1H),5.30(p,J=6.1Hz,1H),4.16(t,J=6.1Hz,2H),3.91(t,J=5.9Hz,2H),2.26–2.14(m,2H),1.89(s,3H),1.30(d,J=6.3Hz,6H).MS(m / z):[M+H] + 503.2.

[0186] Example 22

[0187] 2-((4-(2-(3-Hydroxy-3-(thiazol-2-yl)but-1-yn-1-yl)-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-yl)amino)isonicotinic acid (Compound 11)

[0188] Using the preparation method of Example 7, 2-bromopyridine was replaced with 2-bromoisonicotinic acid to obtain 165 mg of Compound 11 (white solid), with a yield of 87.4%. 1 H NMR(300MHz,DMSO-d 6 )δ10.58(s,1H),8.58(t,J=1.2Hz,1H),8.52(dd,J=5.0,0.9Hz,1H),8.36(d,J=6.0Hz,1H),7.79(d,J=3.2Hz,1H),7.70(d,J=3.2Hz,1H),7.38–7.33(m,2H),7.06(s,1H),6.75(d,J=6.1Hz,1H),4.16(t,J=5.9Hz,2H),3.90(t,J=5.8Hz,2H),2.30-2.12(m,2H),1.89(s,3H).MS(m / z):[M+H] + 470.1.

[0189] Example 23

[0190] 4-(4-(2-((4-(Dimethylamino)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 12)

[0191] Using the preparation method of Example 7, replace 2-bromopyridine with 2-bromo-N,N-dimethylpyridin-4-amine, and keep other processes unchanged to obtain 137 mg of compound 12 (white solid) with a yield of 88.6%. 1 H NMR(300MHz,DMSO-d 6 )δ9.43(s,1H),8.27(d,J=5.9Hz,1H),7.90(d,J=5.9Hz,1H),7.79(d,J=3.3Hz,1H),7.70(d,J=3.2Hz,1H),7.43(d,J=2.4Hz,1H),7.14(s,1H),7.07(s,1H),6.65(d,J=6.0Hz,1H),6.29(dd,J=6.1,2.4Hz,1H),4.15(t,J=6.0Hz,2H),3.93(t,J=5.8Hz,2H),2.95(s,6H),2.28-2.07(m,2H),1.88(s,3H).MS(m / z):[M+H] + 488.2.

[0192] Example 24

[0193] 4-(4-(2-((6-Morpholinopyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 13)

[0194] Using the preparation method of Example 7, replace 2-bromopyridine with 4-(6-bromopyridin-2-yl)morpholine, and keep other processes unchanged to obtain 152 mg of compound 13 (white solid) with a yield of 86.6%. 1 H NMR(300MHz,DMSO-d 6 )δ9.30(s,1H),8.26(d,J=6.1Hz,1H),7.79(d,J=3.3Hz,1H),7.70(d,J=3.2Hz,1H),7.60–7.42(m,2H),7.08(d,J=5.6Hz,2H),6.65(d,J=6.0Hz,1H),6.37(d,J=7.7Hz,1H),4.15(t,J=5.9Hz,2H),3.90(t,J=6.0Hz,2H),3.70(t,J=5.0Hz,4H),3.45(t,J=4.9Hz,7H),2.21(d,J=5.9Hz,2H),1.88(s,3H).MS(m / z):[M+H] + 530.2.

[0195] Example 25

[0196] 4-(4-(2-((6-((2-Methoxyethyl)(methyl)amino)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 14)

[0197] Using the preparation method of Example 7, replace 2-bromopyridine with 6-bromo-N-(2-methoxyethyl)-N-methylpyridin-2-amine, and keep other processes unchanged. 141 mg of Compound 14 (white solid) was obtained with a yield of 84.8%. 1 H NMR(300MHz,DMSO-d 6 )δ9.14(s,1H),8.24(d,J=6.0Hz,1H),7.78(d,J=3.3Hz,1H),7.70(d,J=3.3Hz,1H),7.46-7.36(m,2H),7.12-7.01(m,2H),6.64(d,J=6.1Hz,1H),6.17(dd,J=6.0,2.9Hz,1H),4.14(t,J=5.7Hz,2H),3.90(t,J=5.7Hz,2H),3.70(t,J=5.7Hz,2H),3.50(t,J=5.8Hz,2H),3.26(s,3H),3.00(s,3H),2.25–2.12(m,2H),1.87(s,3H).MS(m / z):[M+H] + 532.2.

[0198] Example 26

[0199] 4-(4-(2-((6-(Piperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 15)

[0200] Using the preparation method of Example 7, replace 2-bromopyridine with 2-bromo-6-(piperidin-1-yl)pyridine, and keep other processes unchanged. A white solid (Compound 15) was obtained in a similar method to the preparation of Compound 7, and 147 mg of the white solid (Compound 15) was obtained with a yield of 88.9%. 1 H NMR(300MHz,DMSO-d 6)δ9.22(s,1H),8.25(d,J=6.0Hz,1H),7.79(d,J=3.3Hz,1H),7.70(d,J=3.3Hz,1H),7.49–7.39(m,2H),7.11(s,1H),7.07(s,1H),6.64(d,J=6.1Hz,1H),6.35(dd,J=7.7,1.3Hz,1H),4.15(t,J=6.0Hz,2H),3.90(t,J=5.9Hz,2H),3.55–3.47(m,4H),2.25-2.14(m,2H),1.88(s,3H),1.64-1.51(m,6H).MS(m / z):[M+H] + 528.2.

[0201] Example 27

[0202] 2-(Thiazol-2-yl)-4-(4-(2-((6-(trifluoromethyl)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)but-3-yn-2-ol (Compound 16)

[0203] Using the preparation method of Example 7, replace 2-bromopyridine with 2-bromo-6-(trifluoromethyl)pyridine, and keep other processes unchanged to obtain 137 mg of Compound 16 (white solid) with a yield of 85.3%. 1 H NMR(300MHz,DMSO-d 6 )δ10.53(s,1H),8.57(d,J=8.6Hz,1H),8.32(d,J=6.0Hz,1H),7.98(t,J=8.1Hz,1H),7.79(d,J=3.3Hz,1H),7.70(d,J=3.2Hz,1H),7.41(d,J=7.0Hz,1H),7.31(s,1H),7.07(s,1H),6.74(d,J=6.1Hz,1H),4.15(t,J=6.0Hz,2H),3.90(t,J=6.0Hz,2H),2.27–2.15(m,2H),1.89(s,3H).MS(m / z):[M+H] + 513.1.

[0204] Example 28

[0205] 4-(4-(2-((5-Fluoropyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 17)

[0206] Using the preparation method of Example 7, replace 2-bromopyridine with 2-bromo-5-fluoropyridine, and keep other processes unchanged. 156 mg of compound 17 (white solid) was obtained with a yield of 80.7%. 1 H NMR(300MHz,DMSO-d 6 )δ10.05(s,1H),8.33-8.22(m,2H),8.17(dd,J=9.2,4.1Hz,1H),7.77(d,J=3.2Hz,1H),7.72–7.62(m,2H),7.26(s,1H),7.06(s,1H),6.66(d,J=6.1Hz,1H),4.14(t,J=6.0Hz,2H),3.88(t,J=5.8Hz,2H),2.25-2.13(m,2H),1.88(s,3H).MS(m / z):[M+H] + 463.1.

[0207] Example 29

[0208] 4-(4-(2-((4-chloropyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 18)

[0209] Using the preparation method of Example 7, replace 2-bromopyridine with 2-bromo-4-chloropyridine, and keep other processes unchanged. 142 mg of compound 18 (white solid) was obtained with a yield of 81.9%. 1 H NMR(300MHz,DMSO-d 6 )δ10.31(s,1H),8.38–8.30(m,2H),8.26(d,J=5.4Hz,1H),7.78(d,J=3.2Hz,1H),7.69(d,J=3.3Hz,1H),7.31(s,1H),7.08–7.02(m,2H),6.71(d,J=6.1Hz,1H),4.14(t,J=6.0Hz,2H),3.89(t,J=5.9Hz,2H),2.26-2.13(m,2H),1.88(s,3H).MS(m / z):[M+H] + 479.1.

[0210] Example 30

[0211] 4-(4-(2-(pyrimidin-4-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 19)

[0212] Using the preparation method of Example 7, replace 2-bromopyridine with 4-bromopyrimidine, and keep other processes unchanged. 172 mg of compound 19 (white solid) was obtained with a yield of 83.3%. 1 H NMR(300MHz,DMSO-d 6 )δ10.62(s,1H),8.79(d,J=1.5Hz,1H),8.53(d,J=6.0Hz,1H),8.36(d,J=6.1Hz,1H),8.25(dd,J=5.9,1.3Hz,1H),7.79(d,J=3.2Hz,1H),7.70(d,J=3.2Hz,1H),7.36(s,1H),7.07(s,1H),6.81(d,J=6.1Hz,1H),4.16(t,J=6.1Hz,2H),3.91(t,J=5.5Hz,2H),2.26–2.14(m,2H),1.89(s,3H).MS(m / z):[M+H] + 446.1.

[0213] Example 31

[0214] 4-(4-(2-(Quinolin-2-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 20)

[0215] Using the preparation method of Example 7, replace 2-bromopyridine with 2-bromoquinoline, and keep other processes unchanged. 152 mg of compound 20 (white solid) was obtained with a yield of 86.6%. 1 H NMR(300MHz,DMSO-d 6 )δ10.34(s,1H),8.41(d,J=9.1Hz,1H),8.33(d,J=6.1Hz,1H),8.26(d,J=9.2Hz,1H),7.85(dd,J=8.0,1.5Hz,1H),7.82–7.76(m,2H),7.75–7.59(m,2H),7.41(ddd,J=8.1,6.8,1.3Hz,1H),7.36(s,1H),7.07(s,1H),6.73(d,J=6.1Hz,1H),4.16(t,J=5.9Hz,2H),3.91(t,J=5.7Hz,2H),2.29–2.14(m,2H),1.87(s,3H).MS(m / z):[M+H] + 495.2.

[0216] Example 32

[0217] 4-(4-(2-(Phenylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 21)

[0218] Using the preparation method of Example 6, replace I-d with I-g-1, and keep other processes unchanged. 155 mg of Compound 21 (white solid) was obtained with a yield of 85.7%. 1 H NMR(400MHz,DMSO-d 6 )δ9.40(s,1H),8.22(d,J=6.0Hz,1H),7.79(d,J=3.3Hz,1H),7.74–7.67(m,3H),7.31–7.23(m,2H),7.05(s,1H),6.97–6.88(m,2H),6.59(d,J=6.0Hz,1H),4.15(t,J=6.1Hz,2H),3.90(t,J=5.8Hz,2H),2.24-2.15(m,2H),1.89(s,3H).MS(m / z):[M+H] + 444.2.

[0219] Example 33

[0220] 4-(4-(2-((3-Methoxyphenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 22)

[0221] Using the preparation method of Example 6, replace I-d with 4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)-N-(3-methoxyphenyl)pyrimidin-2-amine, and keep other processes unchanged. 122 mg of Compound 22 (white solid) was obtained with a yield of 87.8%. 1 H NMR(400MHz,DMSO-d 6)δ9.43(s,1H),8.23(d,J=5.9Hz,1H),7.79(d,J=3.3Hz,1H),7.70(d,J=3.3Hz,1H),7.42(t,J=2.3Hz,1H),7.31(d,J=8.4Hz,1H),7.16(t,J=8.1Hz,1H),7.08(s,1H),6.94(s,1H),6.61(d,J=6.0Hz,1H),6.51(ddd,J=8.1,2.6,0.9Hz,1H),4.15(t,J=6.1Hz,2H),3.91(t,J=5.9Hz,2H),3.73(s,3H),2.24-2.14(m,2H),1.88(s,3H).MS(m / z):[M+H] + 374.2.

[0222] Example 34

[0223] 3-((4-(2-(3-Hydroxy-3-(thiazol-2-yl)but-1-yn-1-yl)-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-yl)amino)benzonitrile (Compound 23)

[0224] Using the preparation method of Example 6, replace I-d with 3-((4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-yl)amino)benzonitrile, and keep other processes unchanged. 145 mg of Compound 23 (white solid) was obtained with a yield of 88.0%. 1 H NMR(300MHz,Chloroform-d)δ9.79(s,1H),8.33–8.26(m,2H),8.00(ddd,J=8.4,2.3,1.1Hz,1H),7.79(d,J=3.2Hz,1H),7.71(d,J=3.2Hz,1H),7.49(t,J=8.0Hz,1H),7.37(dt,J=7.5,1.2Hz,1H),7.07(s,1H),6.96(s,1H),6.70(d,J=6.1Hz,1H),4.16(t,J=6.0Hz,2H),3.92(t,J=5.9Hz,2H),2.26-2.15(m,,2H),1.88(s,3H).MS(m / z):[M+H] + 469.1.

[0225] Example 35

[0226] 4-(4-(2-((3-(Dimethylamino)phenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 24)

[0227] Using the preparation method of Example 6, replace I-d with N1-(4-(2-Iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-yl)-N3,N3-dimethylbenzene-1,3-diamine, and keep other processes unchanged. 149 mg of Compound 24 (white solid) was obtained with a yield of 83.9%. 1 H NMR(300MHz,DMSO-d 6 )δ9.22(s,1H),8.20(d,J=5.8Hz,1H),7.79(d,J=3.3Hz,1H),7.70(d,J=3.2Hz,1H),7.16–7.02(m,4H),6.92(s,1H),6.57(d,J=6.0Hz,1H),6.38–6.25(m,1H),4.15(t,J=6.1Hz,2H),3.91(t,J=5.8Hz,2H),2.87(s,6H),2.24-2.14(m,2H),1.88(s,3H).MS(m / z):[M+H] + 487.2.

[0228] Example 36

[0229] 4-(4-(2-((3-Morpholinophenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 25)

[0230] Using the preparation method of Example 6, replace I-d with 4-(2-Iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)-N-(3-morpholinophenyl)pyrimidin-2-amine, and keep other processes unchanged. 166 mg of Compound 25 (white solid) was obtained with a yield of 83.5%. 1 H NMR(300MHz,DMSO-d 6)δ9.28(s,1H),8.21(d,J=5.9Hz,1H),7.79(d,J=3.2Hz,1H),7.70(d,J=3.2Hz,1H),7.33(t,J=2.2Hz,1H),7.27–7.20(m,1H),7.15 - 7.03(m,2H),6.90(s,1H),6.58(d,J=6.1Hz,1H),6.54(dd,J=7.8,2.8Hz,1H),4.15(t,J=6.1Hz,2H),3.91(t,J=5.8Hz,2H),3.74(dd,J=5.8,3.9Hz,4H),3.10–3.03(m,4H),2.24 - 2.14(m,2H),1.88(s,3H).MS(m / z):[M + H] + 529.2.

[0231] Example 37

[0232] 4-(4-(2-((3,5-Dimethoxyphenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 26)

[0233] Using the preparation method of Example 6, replace I-d with N-(3,5-dimethoxyphenyl)-4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-amine, and keep other processes unchanged. 178 mg of Compound 26 (white solid) was obtained with a yield of 82.8%. 1 H NMR(300MHz,DMSO-d 6 )δ9.41(s,1H),8.23(d,J=6.0Hz,1H),7.78(d,J=3.2Hz,1H),7.70(d,J=3.3Hz,1H),7.08(s,1H),7.04(d,J=2.3Hz,2H),6.95(s,1H),6.62(d,J=6.0Hz,1H),6.10(t,J=2.2Hz,1H),4.15(t,J=6.0Hz,2H),3.92(t,J=5.5Hz,2H),3.71(s,6H),2.25 - 2.14(m,2H),1.87(s,3H).MS(m / z):[M + H] + 503.2.

[0234] Example 38

[0235] 4-(4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 27)

[0236] Using the preparation method of Example 6, replace I-d with N-(benzo[d][1,3]dioxol-5-yl)-4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-amine, and keep other processes unchanged. 154 mg of Compound 27 (white solid) was obtained with a yield of 81.5%. 1 H NMR(300MHz,DMSO-d 6 )δ9.29(s,1H),8.19(d,J=6.0Hz,1H),7.78(d,J=3.3Hz,1H),7.69(d,J=3.2Hz,1H),7.40(d,J=2.1Hz,1H),7.10(dd,J=8.5,2.2Hz,1H),7.06(s,1H),6.90(s,1H),6.83(d,J=8.4Hz,1H),6.55(d,J=6.1Hz,1H),5.97(s,2H),4.14(t,J=6.1Hz,2H),3.89(t,J=5.6Hz,2H),2.25-2.13(m,2H),1.87(s,3H).MS(m / z):[M+H] + 487.1.

[0237] Example 39

[0238] 4-(4-(2-((2,2-Difluorobenzo[d][1,3]dioxol-5-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 28)

[0239] Using the preparation method of Example 6, replace I-d with N-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-amine, and keep other processes unchanged. 182 mg of Compound 28 (white solid) was obtained with a yield of 80.3%. 1 H NMR(300MHz,DMSO-d 6)δ9.63(s,1H),8.24(d,J=6.0Hz,1H),7.94(d,J=2.1Hz,1H),7.78(d,J=3.2Hz,1H),7.69(d,J=3.2Hz,1H),7.44(dd,J=8.9,2.2Hz,1H),7.30(d,J=8.8Hz,1H),7.06(s,1H),6.94(s,1H),6.64(d,J=6.0Hz,1H),4.15(t,J=6.1Hz,2H),3.91(t,J=5.8Hz,2H),2.26 - 2.12(m,2H),1.88(s,3H).MS(m / z):[M + H] + 524.1.

[0240] Example 40

[0241] 4-(4-(2-((4-Fluorophenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 29)

[0242] Using the preparation method of Example 6, replace I-d with N-(4-fluorophenyl)-4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-amine, and keep other processes unchanged, to obtain 163 mg of Compound 29 (white solid) with a yield of 84.3%. 1 H NMR(300MHz,DMSO-d 6 )δ9.45(s,1H),8.21(d,J=6.0Hz,1H),7.78(d,J=3.2Hz,1H),7.75–7.68(m,3H),7.16–7.08(m,2H),7.06(s,1H),6.92(s,1H),6.59(d,J=6.1Hz,1H),4.15(t,J=6.0Hz,2H),3.89(t,J=5.8Hz,2H),2.25–2.13(m,2H),1.88(s,3H).MS(m / z):[M + H] + 462.1.

[0243] Example 41

[0244] 4-(4-(2-((4-Chlorophenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 30)

[0245] Using the preparation method of Example 6, replace Ⅰ-d with N-(4-chlorophenyl)-4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-amine, and keep other processes unchanged. 144 mg of compound 30 (white solid) was obtained with a yield of 85.5%. 1 H NMR(300MHz,DMSO-d 6 )δ9.58(s,1H),8.24(d,J=6.0Hz,1H),7.83–7.75(m,3H),7.69(d,J=3.2Hz,1H),7.35–7.28(m,2H),7.07(s,1H),6.95(s,1H),6.63(d,J=6.1Hz,1H),4.15(t,J=6.0Hz,2H),3.90(t,J=6.0Hz,2H),2.25-2.14(m,2H),1.88(s,3H).MS(m / z):[M+H] + 478.1.

[0246] Example 42

[0247] 4-(4-(2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 31)

[0248] Using the preparation method of Example 6, replace Ⅰ-d with 4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)-N-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-amine, and keep other processes unchanged. 151 mg of compound 31 (white solid) was obtained with a yield of 82.6%. 1 H NMR(300MHz,DMSO-d 6 )δ9.28(s,1H),8.19(d,J=6.0Hz,1H),7.39(d,J=2.1Hz,1H),7.11(dd,J=8.4,2.1Hz,1H),6.83(d,J=8.4Hz,2H),6.56(d,J=6.0Hz,1H),5.97(s,2H),5.33(s,1H),4.13(t,J=6.0Hz,2H),3.89(t,J=5.9Hz,2H),2.22-2.12(m,2H),1.95-1.80(m,4H),1.78–1.64(m,4H).MS(m / z):[M+H] + 448.2.

[0249] Example 43

[0250] 4-(4-(2-((1-Isopropyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 32)

[0251] Using the preparation method of Example 6, replace I-d with 4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)-N-(1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-amine, and keep other processes unchanged, to obtain 188 mg of Compound 32 (white solid) with a yield of 85.1%. 1 H NMR(300MHz,DMSO-d 6 )δ9.24(s,1H),8.17(d,J=6.2Hz,1H),7.89(s,1H),7.77(d,J=3.3Hz,1H),7.69(d,J=3.2Hz,1H),7.45(s,1H),7.30-6.58(m,2H),6.47(d,J=6.0Hz,1H),4.50-4.32(m,1H),4.14(t,J=6.0Hz,2H),3.88(t,J=5.5Hz,2H),2.25-2.12(m,2H),1.87(s,3H),1.38(d,J=6.7Hz,6H).MS(m / z):[M+H] + 476.2.

[0252] Example 44

[0253] 4-(4-(2-((1-Methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 33)

[0254] Using the preparation method of Example 6, replace I-d with 4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)-N-(1-methyl-1H-pyrazol-3-yl)pyrimidin-2-amine, and keep other processes unchanged, to obtain 140 mg of Compound 33 (white solid) with a yield of 89.1%. 1 H NMR(300MHz,DMSO-d 6)δ9.55(s,1H),8.16(d,J=6.0Hz,1H),7.80(d,J=3.2Hz,1H),7.70(d,J=3.2Hz,1H),7.49(d,J=2.2Hz,1H),7.11(s,1H),7.06(s,1H),6.49(d,J=6.0Hz,1H),6.39(s,1H),4.13(t,J=6.1Hz,2H),3.85(t,J=6.0Hz,2H),3.74(s,3H),2.23 - 2.12(m,2H),1.88(s,3H).MS(m / z):[M + H] + 447.2.

[0255] Example 45

[0256] 4-(4-((4-Fluorophenyl)amino)-1,3,5-triazin-2-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 34)

[0257] Using the preparation method of Example 6, replace I-d with N-(4-fluorophenyl)-4-(2-iodo-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)-1,3,5-triazin-2-amine, and keep other processes unchanged, to obtain 139 mg of Compound 34 (white solid), with a yield of 86.4%. 1 H NMR(300MHz,DMSO-d 6 )δ10.06(s,1H),8.50(s,1H),7.78(d,J=3.2Hz,1H),7.73–7.56(m,4H),7.19(dd,J=9.9,7.8Hz,2H),7.07(s,1H),4.15(q,J=7.1,6.6Hz,4H),2.18(s,2H),1.88(s,3H).MS(m / z):[M + H] + 362.1.

[0258] Example 46

[0259] 4-(4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-methylbut-3-yn-2-ol (Compound 35)

[0260] Using the preparation method of Example 6, replace I-d with I-h, replace 2-(thiazol-2-yl)but-3-yn-2-ol with 2-methylbut-3-yn-2-ol, and keep other processes unchanged. 122 mg of compound 35 (white solid) was obtained with a yield of 85.7%. 1 HNMR(300MHz,DMSO-d 6 )δ9.24(s,1H),8.17(d,J=5.9Hz,1H),7.37(d,J=2.2Hz,1H),7.09(dd,J=8.4,2.2Hz,1H),6.82(d,J=8.4Hz,2H),6.53(d,J=6.1Hz,1H),5.96(s,2H),5.44(s,1H),4.12(t,J=5.9Hz,2H),3.87(t,J=5.9Hz,2H),2.23-2.09(m,2H),1.45(s,6H).MS(m / z):[M+H] + 418.2.

[0261] Example 47

[0262] 3-((4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)ethynyl)oxetan-3-ol (Compound 36)

[0263] Using the preparation method of Example 6, replace I-d with I-h, replace 2-(thiazol-2-yl)but-3-yn-2-ol with 3-ethynyloxetan-3-ol, and keep other processes unchanged. 142 mg of compound 36 (white solid) was obtained with a yield of 83.3%. 1 H NMR(300MHz,DMSO-d 6 )δ9.27(s,1H),8.20(d,J=5.9Hz,1H),7.40(d,J=2.2Hz,1H),7.09(dd,J=8.5,2.2Hz,1H),6.95(s,1H),6.84(d,J=8.4Hz,1H),6.65(s,1H),6.55(d,J=6.0Hz,1H),5.97(s,2H),4.73(d,J=6.9Hz,2H),4.61(d,J=6.8Hz,2H),4.16(t,J=6.0Hz,2H),3.89(t,J=5.9Hz,2H),2.26-2.12(m,2H).MS(m / z):[M+H] + 432.2.

[0264] Example 48

[0265] 1-((4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)ethynyl)cyclopent-1-ol (Compound 37)

[0266] Using the preparation method of Example 6, replace I-d with I-h, replace 2-(thiazol-2-yl)but-3-yn-2-ol with 1-ethynylcyclopent-1-ol, and keep other processes unchanged to obtain 176 mg of Compound 37 (white solid) with a yield of 84.7%. 1 H NMR(300MHz,DMSO-d 6 )δ9.28(s,1H),8.19(d,J=6.0Hz,1H),7.39(d,J=2.1Hz,1H),7.11(dd,J=8.4,2.1Hz,1H),6.83(d,J=8.4Hz,2H),6.56(d,J=6.0Hz,1H),5.97(s,2H),5.33(s,1H),4.13(t,J=6.0Hz,2H),3.89(t,J=5.9Hz,2H),2.22-2.12(m,2H),1.95-1.80(m,4H),1.78–1.64(m,4H).MS(m / z):[M+H] + 444.2.

[0267] Example 49

[0268] 7-((4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)ethynyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-7-ol (Compound 38)

[0269] Using the preparation method of Example 6, replace I-d with I-h, replace 2-(thiazol-2-yl)but-3-yn-2-ol with 7-ethynyl-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-7-ol, and keep other processes unchanged to obtain 171 mg of Compound 38 (white solid) with a yield of 79.3%. 1 H NMR(300MHz,DMSO-d 6)δ9.26(s,1H),8.18(s,1H),7.40(s,1H),7.23–6.86(m,4H),6.82(d,J=8.2Hz,1H),6.59-6.42(m,2H),5.95(d,J=4.1Hz,2H),4.14(t,J=6.0Hz,2H),4.10-3.96(m,2H),3.87(t,J=5.6Hz,2H),3.05-2.91(m,1H),2.86-2.70(m,1H),2.28-2.12(m,2H).MS(m / z):[M+H] + 483.2.

[0270] Example 50

[0271] 4-(4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(5-methylisoxazol-3-yl)but-3-yn-2-ol (Compound 39)

[0272] Using the preparation method of Example 6, replace I-d with I-h, replace 2-(thiazol-2-yl)but-3-yn-2-ol with 2-(5-methylisoxazol-3-yl)but-3-yn-2-ol, and keep other processes unchanged to obtain 128 mg of Compound 39 (white solid) with a yield of 84.9%. 1 H NMR(300MHz,DMSO-d 6 )δ9.28(s,1H),8.18(d,J=6.0Hz,1H),7.39(d,J=2.1Hz,1H),7.10(dd,J=8.4,2.1Hz,1H),6.89(s,1H),6.83(d,J=8.3Hz,1H),6.60–6.48(m,2H),6.32(d,J=1.0Hz,1H),5.96(s,2H),4.14(t,J=6.1Hz,2H),3.88(t,J=5.8Hz,2H),2.41(d,J=1.0Hz,3H),2.25-2.12(m,2H),1.79(s,3H).MS(m / z):[M+H] + 486.2.

[0273] Example 51

[0274] 4-(7-(2-Aminopyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 40)

[0275] Using the preparation method of Example 6, replace Ⅰ-d with Ⅰ-n prepared in Example 16, and keep other processes unchanged. 139 mg of compound 40 (white solid) was obtained with a yield of 86.1%. 1 H NMR(300MHz,DMSO-d 6 )δ8.43(s,1H),8.15(d,J=7.3Hz,1H),7.92–7.70(m,3H),7.61(dq,J=13.2,7.6,7.1Hz,2H),7.50–7.29(m,4H),6.78(s,2H),1.93(s,3H).MS(m / z):[M+H] + 363.1.

[0276] Example 52

[0277] 4-(7-(2-(Bicyclo[1.1.1]pentan-1-ylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 41)

[0278] Using the preparation method of Example 6, replace Ⅰ-d with N-(bicyclo[1.1.1]pentan-1-yl)-4-(2-iodobenzofuran-7-yl)pyrimidin-2-amine, and keep other processes unchanged. 152 mg of compound 41 (white solid) was obtained with a yield of 87.3%. 1 H NMR(300MHz,DMSO-d 6 )δ8.48(d,J=5.1Hz,1H),8.21(d,J=7.9Hz,1H),7.98(s,1H),7.85–7.78(m,2H),7.74(d,J=3.3Hz,1H),7.53–7.40(m,3H),7.38(s,1H),2.47(s,1H),2.14(s,6H),1.93(s,3H).MS(m / z):[M+H] + 429.1

[0279] Example 53

[0280] 4-(7-(2-((1-Methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 42)

[0281] Using the preparation method of Example 6, replace Ⅰ-d with 4-(2-iodobenzofuran-7-yl)-N-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-amine, and keep other processes unchanged. 102 mg of compound 42 (white solid) was obtained with a yield of 88.2%. 11H NMR (300 MHz, DMSO-d 6 ) δ 9.62 (s, 1H), 8.61 (d, J = 5.1 Hz, 1H), 8.20 (dd, J = 7.7, 1.3 Hz, 1H), 7.98 (s, 1H), 7.89–7.79 (m, 2H), 7.74 (d, J = 3.2 Hz, 1H), 7.62 (s, 1H), 7.56–7.39 (m, 4H), 3.83 (s, 3H), 1.93 (s, 3H). MS (m / z): [M+H] + 443.1.

[0282] Example 54

[0283] 4-(7-(2-((1-Methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 43)

[0284] Using the preparation method of Example 6, replace I-d with 4-(2-iodobenzofuran-7-yl)-N-(1-methyl-1H-pyrazol-3-yl)pyrimidin-2-amine, and keep other processes unchanged, to obtain 131 mg of Compound 43 (white solid), with a yield of 82.6%. 1 1H NMR (300 MHz, DMSO-d 6 ) δ 9.91 (s, 1H), 8.63 (d, J = 5.1 Hz, 1H), 8.23 (dd, J = 7.7, 1.4 Hz, 1H), 7.90–7.80 (m, 2H), 7.76 (d, J = 3.3 Hz, 1H), 7.63–7.40 (m, 5H), 6.78 (d, J = 2.1 Hz, 1H), 3.77 (s, 3H), 1.94 (s, 3H). MS (m / z): [M+H] + 443.1.

[0285] Example 55

[0286] 4-(7-(2-((1-(Tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 44)

[0287] Using the preparation method of Example 6, replace I-d with 4-(2-iodobenzofuran-7-yl)-N-(1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)pyrimidin-2-amine, and keep other processes unchanged, to obtain 138 mg of Compound 44 (white solid), with a yield of 84.2%. 1 1H NMR (400 MHz, DMSO-d6 ) δ 9.61 (d, J = 10.4 Hz, 1H), δ 8.61 (d, J = 5.3 Hz, 1H), 8.27–8.14 (m, 1H), 8.02 (s, 1H), 7.90–7.79 (m, 2H), 7.74 (d, J = 3.3 Hz, 2H), 7.61–7.37 (m, 4H), 4.39 (td, J = 10.9, 5.4 Hz, 1H), 3.97 (tt, J = 6.9, 3.9 Hz, 2H), 3.47 (tt, J = 11.3, 3.7 Hz, 2H), 1.95 (d, J = 14.0 Hz, 7H). MS (m / z): [M+H] + 513.2.

[0288] Example 56

[0289] 4-(7-(2-((1-(2-(Dimethylamino)ethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 45)

[0290] Using the preparation method of Example 6, replace I-d with N-(1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)-4-(2-iodobenzofuran-7-yl)pyrimidin-2-amine, and keep other processes unchanged, 102 mg of Compound 45 (white solid) was obtained with a yield of 84.3%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.59 (s, 1H), 8.61 (d, J = 5.1 Hz, 1H), 8.22 (d, J = 6.4 Hz, 1H), 8.03 (s, 1H), 7.88–7.79 (m, 2H), 7.74 (d, J = 3.2 Hz, 1H), 7.64 (s, 1H), 7.56–7.38 (m, 4H), 4.18 (t, J = 6.5 Hz, 2H), 2.64 (t, J = 6.5 Hz, 2H), 2.17 (s, 6H), 1.94 (s, 3H), 1.00 (s, 3H). MS (m / z): [M+H] + 500.1.

[0291] Example 57

[0292] 4-(7-(2-((1-Methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 46)

[0293] Using the preparation method of Example 6, replace I-d with 4-(2-iodobenzofuran-7-yl)-N-(1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)pyrimidin-2-amine, and keep other processes unchanged. 148 mg of compound 46 (white solid) was obtained with a yield of 84.7%. 1 H NMR(400MHz,Chloroform-d)δ8.61(d,J=4.8Hz,1H),8.43–8.19(m,2H),7.96–7.76(m,2H),7.69(d,J=7.8Hz,1H),7.50–7.37(m,2H),7.33(s,2H),7.08(s,1H),3.95(s,3H),2.13(s,3H).MS(m / z):[M+H] + 511.1.

[0294] Example 58

[0295] 4-(7-(2-(phenylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 47)

[0296] Using the preparation method of Example 6, replace I-d with 4-(2-iodobenzofuran-7-yl)-N-phenylpyrimidin-2-amine, and keep other processes unchanged. 133 mg of compound 47 (white solid) was obtained with a yield of 80.7%. 1 H NMR(300MHz,DMSO-d 6 )δ9.78(s,1H),8.68(d,J=5.1Hz,1H),8.22(dd,J=7.7,1.4Hz,1H),7.93–7.80(m,4H),7.75(d,J=3.3Hz,1H),7.60(d,J=5.1Hz,1H),7.56–7.45(m,2H),7.40(s,1H),7.37–7.28(m,2H),7.00–6.91(m,1H),1.93(s,3H).MS(m / z):[M+H] + 439.1.

[0297] Example 59

[0298] 4-(7-(2-((3-methoxyphenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 48)

[0299] Using the preparation method of Example 6, replace I-d with 4-(2-iodobenzofuran-7-yl)-N-(3-methoxyphenyl)pyrimidin-2-amine, and keep other processes unchanged to obtain 130 mg of Compound 48 (white solid) with a yield of 85.1%. 1 H NMR(300MHz,DMSO-d 6 )δ9.80(s,1H),8.68(s,1H),8.25(d,J=7.7Hz,1H),7.88–7.79(m,2H),7.74(d,J=3.4Hz,1H),7.64–7.58(m,2H),7.54–7.38(m,4H),7.22(t,J=8.2Hz,1H),6.54(dd,J=8.1,2.7Hz,1H),3.75(s,3H),1.93(s,3H).MS(m / z):[M+H] + 469.1.

[0300] Example 60

[0301] 3-((4-(2-(3-Hydroxy-3-(thiazol-2-yl)but-1-yn-1-yl)benzofuran-7-yl)pyrimidin-2-yl)amino)benzonitrile (Compound 49)

[0302] Using the preparation method of Example 6, replace I-d with 3-((4-(2-iodobenzofuran-7-yl)pyrimidin-2-yl)amino)benzonitrile, and keep other processes unchanged to obtain 121 mg of Compound 49 (white solid) with a yield of 80.8%. 1 H NMR(300MHz,DMSO-d 6 )δ10.19(s,1H),8.75(s,1H),8.55–8.09(m,3H),8.09–7.17(m,9H),1.93(s,3H).MS(m / z):[M+H] + 464.1.

[0303] Example 61

[0304] 4-(7-(2-((3-(Dimethylamino)phenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 50)

[0305] Using the preparation method of Example 6, replace I-d with N1-(4-(2-iodobenzofuran-7-yl)pyrimidin-2-yl)-N3,N3-dimethylbenzene-1,3-diamine, and keep other processes unchanged to obtain 147 mg of Compound 50 (white solid) with a yield of 83.5%. 11H NMR (300 MHz, DMSO-d 6 ) δ 9.57 (s, 1H), 8.67 (d, J = 5.1 Hz, 1H), 8.35–8.24 (m, 1H), 7.84 (dd, J = 8.5, 2.2 Hz, 2H), 7.75 (d, J = 3.2 Hz, 1H), 7.60 (d, J = 5.1 Hz, 1H), 7.56–7.35 (m, 4H), 7.14 (dt, J = 16.0, 8.2 Hz, 2H), 6.38 (dd, J = 5.2, 2.8 Hz, 1H), 2.91 (s, 6H), 1.94 (s, 3H). MS (m / z): [M+H] + 482.2.

[0306] Example 62

[0307] 4-(7-(2-((3-Morpholinophenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 51)

[0308] Using the preparation method of Example 6, replace I-d with 4-(2-iodobenzofuran-7-yl)-N-(3-morpholinophenyl)pyrimidin-2-amine, and keep other processes unchanged to obtain Compound 51 (white solid). 132 mg of white solid (Compound 51) was obtained, with a yield of 87.3%. 1 1H NMR (300 MHz, DMSO-d 6 ) δ 9.62 (s, 1H), 8.67 (d, J = 5.3 Hz, 1H), 8.26 (d, J = 7.9 Hz, 1H), 8.12–6.80 (m, 10H), 6.58 (d, J = 7.8 Hz, 1H), 3.76 (t, J = 4.8 Hz, 4H), 3.11 (t, J = 4.9 Hz, 4H), 1.94 (s, 3H). MS (m / z): [M+H] + 524.2.

[0309] Example 63

[0310] 4-(7-(2-((4-Fluorophenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 52)

[0311] Using the preparation method of Example 6, replace I-d with N-(4-fluorophenyl)-4-(2-iodobenzofuran-7-yl)pyrimidin-2-amine, and keep other processes unchanged to obtain 178 mg of Compound 52 (white solid), with a yield of 89.2%. 1 1H NMR (300 MHz, DMSO-d 6)δ9.81(s,1H),8.67(d,J=5.1Hz,1H),8.19(dd,J=7.7,1.3Hz,1H),7.90(d,J=5.0Hz,1H),7.88–7.83(m,2H),7.81(d,J=3.2Hz,1H),7.74(d,J=3.3Hz,1H),7.59(d,J=5.1Hz,1H),7.51(d,J=7.7Hz,1H),7.47(d,J=1.3Hz,1H),7.40(s,1H),7.22–7.13(m,2H),1.93(s,3H).MS(m / z):[M+H] + 457.1.

[0312] Example 64

[0313] 4-(7-(2-(((4-chlorophenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 53)

[0314] Using the preparation method of Example 6, replace I-d with N-(4-chlorophenyl)-4-(2-iodobenzofuran-7-yl)pyrimidin-2-amine, and keep other processes unchanged to obtain Compound 53 (white solid). 161 mg of white solid (Compound 53) was obtained, with a yield of 85.6%. 1 H NMR(300MHz,DMSO-d 6 )δ9.94(s,1H),8.70(d,J=5.2Hz,1H),8.20(dd,J=7.6,1.3Hz,1H),7.94(d,J=2.0Hz,1H),7.91(d,J=2.1Hz,1H),7.85(dd,J=7.8,1.2Hz,1H),7.82(d,J=3.2Hz,1H),7.74(d,J=3.1Hz,1H),7.62(d,J=5.1Hz,1H),7.51(t,J=7.7Hz,1H),7.47(s,1H),7.40(d,J=1.3Hz,2H),7.37(d,J=1.9Hz,1H),1.94(s,3H).MS(m / z):[M+H] + 472.1.

[0315] Example 65

[0316] 4-(7-(2-(benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 54)

[0317] Using the preparation method of Example 6, replace I-d with N-(benzo[d][1,3]dioxol-5-yl)-4-(2-iodobenzofuran-7-yl)pyrimidin-2-amine, and keep other processes unchanged. 156 mg of compound 54 (white solid) was obtained with a yield of 83.8%. 1 H NMR(300MHz,DMSO-d 6 )δ9.66(s,1H),8.64(d,J=5.2Hz,1H),8.19(dd,J=7.7,1.3Hz,1H),7.87–7.80(m,2H),7.74(d,J=3.2Hz,1H),7.58–7.49(m,3H),7.47(d,J=2.8Hz,1H),7.40(s,1H),7.28(dd,J=8.5,2.2Hz,1H),6.89(d,J=8.4Hz,1H),5.98(s,2H),1.93(s,3H).MS(m / z):[M+H] + 483.1.

[0318] Example 66

[0319] 4-(7-(2-((2,3-dihydrobenzo[b][1,4]dioxan-6-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 55)

[0320] Using the preparation method of Example 6, replace I-d with N-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)-4-(2-iodobenzofuran-7-yl)pyrimidin-2-amine, and keep other processes unchanged. 126 mg of compound 55 (white solid) was obtained with a yield of 84.1%. 1 H NMR(400MHz,Chloroform-d)δ8.51(d,J=5.4Hz,1H),8.27(dd,J=7.7,1.3Hz,1H),7.82(d,J=3.3Hz,1H),7.76(d,J=3.3Hz,1H),7.72(d,J=5.3Hz,1H),7.66(dd,J=7.8,1.4Hz,1H),7.47–7.33(m,5H),7.13(dd,J=8.8,2.6Hz,1H),7.05(s,1H),6.88(d,J=8.6Hz,1H),4.28(tt,J=5.1,2.9Hz,4H),2.13(s,3H).MS(m / z):[M+H] + 496.1.

[0321] Example 67

[0322] 4-(7-(2-(Pyridin-2-ylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 56)

[0323] Using the preparation method of Example 7, replacing Compound 1 with Compound 40 prepared in Example 51, and keeping other processes unchanged, 147 mg of Compound 56 (white solid) was obtained with a yield of 80.9%. 1 H NMR(300MHz,DMSO-d 6 )δ10.00(s,1H),8.76(s,1H),8.26(d,J=7.3Hz,1H),7.98(d,J=7.8Hz,2H),7.89(dd,J=10.9,5.8Hz,2H),7.80(t,J=3.6Hz,1H),7.68(d,J=5.2Hz,1H),7.54(d,J=7.7Hz,2H),7.44(d,J=8.6Hz,3H),1.99(s,3H).MS(m / z):[M+H] + 439.1.

[0324] Example 68

[0325] 4-(7-(2-((6-(Dimethylamino)pyridin-2-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 57)

[0326] Using the preparation method of Example 7, replacing Compound 1 with Compound 40, and replacing 2-bromopyridine with 6-bromo-N,N-dimethylpyridin-2-amine, and keeping other processes unchanged, 138 mg of Compound 57 (white solid) was obtained with a yield of 83.0%. 1 HNMR(400MHz,Chloroform-d)δ8.59(d,J=5.1Hz,1H),8.33–8.13(m,2H),7.89–7.59(m,3H),7.43–7.34(m,2H),7.31(s,1H),7.05(s,1H),4.56–4.25(m,1H),3.92(s,3H),2.11(s,3H).MS(m / z):[M+H] + 483.2.

[0327] Example 69

[0328] 4-(7-(2-((6-Morpholinopyridin-2-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 58)

[0329] Using the preparation method of Example 7, replace compound 1 with compound 40, and react 2-bromopyridine with 4-(6-bromopyridin-2-yl)morpholine. Keep other processes unchanged to obtain 108 mg of compound 58 (white solid) with a yield of 82.2%. 1 HNMR(300MHz,DMSO-d 6 )δ9.34(s,1H),8.73(d,J=5.2Hz,1H),8.26(dd,J=7.7,1.3Hz,1H),7.85(dd,J=7.8,1.2Hz,1H),7.83(d,J=3.2Hz,1H),7.78(d,J=7.9Hz,1H),7.75(d,J=3.2Hz,1H),7.69(d,J=5.2Hz,1H),7.59(t,J=8.1Hz,1H),7.51(t,J=7.7Hz,1H),7.48(s,1H),7.40(s,1H),6.42(d,J=8.2Hz,1H),3.71(dd,J=5.8,3.8Hz,4H),3.46(t,J=4.8Hz,4H),1.93(s,3H).MS(m / z):[M+H] + 525.2.

[0330] Example 70

[0331] 4-(7-(2-((6-Methoxypyridin-2-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 59)

[0332] Using the preparation method of Example 7, replace compound 1 with compound 40, and react 2-bromopyridine with 2-bromo-6-methoxypyridine. Keep other processes unchanged to obtain 129 mg of compound 59 (white solid) with a yield of 80.2%. 1 H NMR(300MHz,DMSO-d 6 )δ9.64(s,1H),8.75(d,J=5.2Hz,1H),8.26(dd,J=7.7,1.3Hz,1H),8.05(d,J=7.4Hz,1H),7.92–7.79(m,2H),7.79–7.67(m,3H),7.58–7.45(m,2H),7.40(s,1H),6.41(dd,J=8.0,0.6Hz,1H),3.87(s,3H),1.93(s,3H).MS(m / z):[M+H] + 470.1.

[0333] Example 71

[0334] 4-(7-(2-((2-(Difluoromethyl)pyridin-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (Compound 60)

[0335] Using the preparation method of Example 7, replacing Compound 1 with Compound 40 and reacting 2-bromopyridine with 4-bromo-2-(difluoromethyl)pyridine, with other processes remaining unchanged, 144 mg of Compound 60 (yellow solid) was obtained with a yield of 84.4%. 1 HNMR(300MHz,DMSO-d 6 )δ10.56(s,1H),8.83(d,J=5.2Hz,1H),8.51(d,J=5.7Hz,1H),8.30–8.23(m,2H),8.03(d,J=5.2Hz,1H),7.88(dd,J=7.8,1.3Hz,1H),7.81(t,J=4.0Hz,2H),7.74(d,J=3.3Hz,1H),7.54(d,J=7.7Hz,1H),7.50(d,J=1.8Hz,1H),7.40(s,1H),6.91(s,1H),1.93(s,3H).MS(m / z):[M+H] + 490.1.

[0336] Example 72

[0337] Experimental study on the binding ability of some compounds to NIK protein: ADP-Glo TM Kinase Assay was used to screen for inhibitors, specifically including the following steps:

[0338] 1) Experimental materials and equipment: Used 25μ1:25μ1:50μ1 in a 96-well plate for a total of 20,000 determinations.

[0339] Including:

[0340] a) 10×50ml ADP-Glo TM Reagent.

[0341] b) 10×100ml kinase detection light yellow.

[0342] c) 10 vials of kinase detection substrate (lyophilized).

[0343] d) 10×5ml ultrapure ATP, 10mM.

[0344] e) 10×5ml ADP, 10mM.

[0345] f) V9104 Component Pack (Helix On-Site Stocking Program).

[0346] 2) Experimental principle:

[0347] ADP-Glo TM The ADP-Glo Kinase Assay is a luminescent ADP detection method that provides a general, homogeneous, high-throughput screening method for measuring kinase activity by quantifying the amount of ADP generated during the kinase reaction.

[0348] ADP-Glo TM The ADP-Glo Kinase Assay can be used to monitor the activity of almost any ADP-generating enzyme (such as kinases or ATPases), using up to 1 mM ATP. ADP-Glo TM The ADP-Glo Kinase Assay is performed in multi-well plates of various different formats and can detect kinase activity in reaction volumes as low as 5 μL. The assay is carried out in two steps: First, after the kinase reaction, an equal volume of ADP-Glo TM reagent is added to terminate the kinase reaction and deplete the remaining ATP. Second, the kinase detection reagent is added, which simultaneously converts ADP to ATP, and the newly synthesized ATP is detected using the luciferase / luciferin reaction. The light generated during the detection process is measured using a luminometer. The luminescence can be correlated to the ADP concentration using an ATP-ADP conversion curve. This assay is sensitive enough to detect very low amounts of ADP (20 nM) and can detect ADP generated in the reaction containing 1 mM ATP in a linear manner; the intensity of the luminescence signal is proportional to the amount of ADP generated and the inhibitory activity of the inhibitor on the kinase.

[0349] 3) Experimental procedures and methods: The following ADP-Glo TM Kinase Assay screening inhibitor operating procedures are taken as an example, and the actual volume can be adjusted according to requirements. ADP-Glo TM The volume of the reagent is the same as the volume of the kinase reaction, and the volume of the kinase detection reagent should be twice the volume of the kinase reaction. This is the reagent volume for a 384-well plate (5 μL: 5 μL: 10 μL); when using a 96-well plate, it is 5 times this volume.

[0350] 1. Add 1 μL of the test compound to each well, and add 1 μL of the solvent of the test compound in another 16 wells as a control

[0351] 2. Add 2 μL of 1× kinase reaction buffer without kinase substrate to 8 control wells.

[0352] 3. Add 2 μL of 1× kinase reaction buffer containing kinase substrate to the remaining wells respectively.

[0353] 4. Add 2 μL of 1× kinase reaction buffer containing 2.5× concentration of ATP (up to 1 mM) to all wells.

[0354] 5. Incubate for 40 minutes after mixing.

[0355] 6. Add 5 μL of ADP-Glo reagent to terminate the kinase reaction and deplete the remaining ATP, leaving only ADP and a small amount of ATP.

[0356] 7. Note: The ADP-Glo reagent can effectively terminate the kinase reaction, so there is no need to add additional inhibitors to terminate the kinase reaction. It is not recommended to modify the experimental procedure to add kinase termination reagents. If a kinase terminator is added, do not use magnesium chelators such as EDTA because the ADP-Glo Assay requires magnesium. NOTE: The final concentration of magnesium ions should be at least 0.5 mM.

[0357] 8. Incubate at room temperature for 40 minutes.

[0358] 9. Add 10 μL of kinase detection reagent to convert ADP to ATP, and detect ATP using luciferase and luciferin.

[0359] 10. Select the incubation time according to the ATP concentration in the kinase reaction (as shown in Table 1), generally incubating for 30 - 60 minutes at room temperature.

[0360] Table 1 Selection of Incubation Time

[0361] ATP concentration 10 - 100 μM 100 - 500 μM 500 - 1000 μM Time 30 minutes 40 minutes 60 minutes

[0362] 11. Measure the fluorescence using a plate spectrophotometer or an electrically coupled camera.

[0363] 12. Set up the instrument according to the manufacturer's instructions. The reference integration time is 0.25 - 1 second per well. If a long half-life signal of the ADP-Glo TM Kinase Assay is required, the plate can be left at room temperature for a longer time before reading.

[0364] 13. Record the fluorescence data. NOTE: Set up the instrument according to the manufacturer's instructions. The reference integration time is 0.25 - 1 second per well.

[0365] For further determination of kinase inhibitor IC 50 is as follows:

[0366] The following operating procedures apply to 96-well plates with a volume ratio of 25 μL:25 μL:50 μL. When using a 384-well plate, reduce the volume by five times. Other volumes can also be used, but for the kinase reaction volume, ADP-Glo TMThe ratio of the reagent volume to the kinase detection reagent volume remains unchanged. The specific operation example is the same as that of ADP-Glo TM Kinase Assay inhibitor screening.

[0367] The following table shows the IC 50 values of the inhibitory activities of some compounds against NIK kinase, and the positive reference is B022:

[0368]

[0369]

[0370]

[0371] As shown in Table 1, the compounds provided by the present invention have a brand-new backbone structure and exhibit a relatively high affinity for NIK protein, and can well inhibit the activity of NIK protein.

[0372] Example 73

[0373] Determination of nitric oxide (NO) release from peritoneal macrophages by Griess reagent method

[0374] 1. Macrophages extracted from the peritoneal cavity of mice were inoculated into 96-well plates (2×10 4 cell / mL, 200 μL per well), and the culture medium (5% fetal bovine serum, 1% penicillin-streptomycin, 1% non-essential amino acids, 2% l-glutamine). Incubate at 37°C, 5% CO 2 for 24 hours.

[0375] 2. After 24 hours, discard the culture medium. Add 197 μL / well of supplemented culture medium 1640. Subsequently, add 2 μL of DMSO-dissolved test compound with or without (1 μM concentration, 3 replicates / sample) to the test drug group and the LPS model control group. For the blank group, only 2 μL of culture medium was added. Incubate the plates in a 5% CO 2 , 37°C incubator for 2 hours.

[0376] 3. After 2 hours, add 1 μL of LPS (200 μg / mL) prepared with PBS to the LPS control group and each test group, with a final concentration of 1 μg / mL, and culture for another 22 hours.

[0377] After 4.22 hours, 100 μL of the supernatant in each well was transferred to a new 96-well plate. Subsequently, 100 μL of Griess reagent (1% sulfanilamide, 0.1% naphthylenediamine dihydrochloride, and 2.5% phosphoric acid) was added to the supernatant. Finally, using linear regression analysis of the standard curve (double dilution of sodium nitrite, from 200 μmol / L to the eleventh dilution), the NO concentration in the supernatant was measured using a microplate reader. The absorbance was measured at 540 nm.

[0378] Table 2 shows the inhibition rates of some compounds on the release of nitric oxide (NO) by peritoneal macrophages

[0379]

[0380] As shown in Table 2, the compounds provided by the present invention exhibited good anti-inflammatory activity and were able to effectively inhibit the production of the inflammatory factor nitric oxide by peritoneal macrophages. The inhibitory effects of some compounds exceeded that of B022, such as compounds 21, 23, 24, and 27.

[0381] Example 74

[0382] ELISA detection of the secretion of macrophage inflammatory factors

[0383] 1. RAW264.7 cells were resuscitated and passaged. After counting, they were seeded into 48-well plates at 1×10 5 cells / well (the medium used was DMEM medium supplemented with 10% FBS)

[0384] 2. After 2 hours, the cells were changed to serum-free DMEM medium, and the treatment group was added with 250 μL of the medium containing 5 μM of the corresponding compound and continued to be cultured

[0385] 3. After adding the compound for 2 hours, cells were stimulated with LPS and CD40 at final concentrations of 100 ng / mL and 500 ng / mL per well.

[0386] 4. After 22 hours, the cell supernatant was aspirated and detected using an ELISA detection kit. The kits used were as follows: TNF-α kit (manufacturer Proteintech, catalog number KE10002), IL-6 kit (manufacturer Proteintech, catalog number KE10007).

[0387] As Figure 1 shown, for the inhibition of the secretion of inflammatory factors by compound 27 in RAW264.7 macrophages, the compounds provided by the present invention exhibited good anti-inflammatory activity and were able to effectively inhibit the secretion of TNF-α and IL-6 inflammatory factors by RAW264.7 macrophages.

[0388] In summary, the compounds provided in Example 72 of the present invention exhibit good NIK protein affinity and can significantly inhibit NIK. The in vitro anti-inflammatory activity evaluations of Examples 73 and 74 also show that the compounds provided by the present invention exhibit good anti-inflammatory activity and can well inhibit the production of nitric oxide by peritoneal macrophages and the secretion of TNF-α and IL-6 inflammatory factors by RAW264.7 macrophages. Since NIK plays a key role in the pathogenesis of inflammatory diseases such as tumors and autoimmune diseases, the compounds of the present invention can be used to prevent or treat diseases such as cancer (e.g., B-cell malignancies, including leukemia, lymphoma, and myeloma), inflammatory diseases, autoimmune diseases, and metabolic diseases such as obesity and diabetes, supported by the in vitro protein inhibition and anti-inflammatory activity experimental data.

Claims

1. A compound of the following formula (I) or a pharmaceutically acceptable salt thereof: Wherein, represents a single bond or a double bond; X 1 , X 4 , X 5 and X 6 are each independently selected from C, N or CH; X 2 Selected from N or CH; X 3 selected from O or CH; R 1 selected from hydrogen or C 1 -C 6 alkyl; R 2 selected from C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, 4-12 membered heterocyclic group or 5-10 membered heteroaryl; R 3 selected from hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 10 cycloalkyl, 4- to 7-membered heterocyclic group, 5- to 10-membered heteroaryl or 5- to 10-membered aryl.

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, When said represents a single bond, X 1 , X 2 and X 4 are N, X 3 is CH, X 5 and X 6 are each independently selected from N or CH.

3. The compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, When said represents a double bond, X 1 is selected from C or N, X 2 and X 3 is selected from CH or O, X 4 is C, X 5 and X 6 are respectively selected from CH or N.

4. The compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, the compound is selected from any one of the following compounds: 4-(4-(2-Aminopyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (1); 4-(4-(2-(Methylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (2); 4-(4-(2-(Isopropylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (3); 4-(4-(2-(2-Methoxyethyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (4); 4-(4-(2-(Cyclohexylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (5); 4-(4-(2-(Bicyclo 1.1.1]pentan-1-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (6); 4-(4-(2-(Pyridin-2-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (7); 4-(4-(2-((4-Methoxypyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (8); 4-(4-(2-((4,6-Dimethoxypyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (9); 4-(4-(2-((6-Isopropoxypyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (10); 2-((4-(2-(3-Hydroxy-3-(thiazol-2-yl)but-1-yn-1-yl)-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-yl)amino)isonicotinic acid (11); 4-(4-(2-((4-(Dimethylamino)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (12); 4-(4-(2-((6-Morpholinopyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (13); 4-(4-(2-((6-((2-Methoxyethyl)(methyl)amino)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (14); 4-(4-(2-((6-(Piperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (15); 2-(Thiazol-2-yl)-4-(4-(2-((6-(Trifluoromethyl)pyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)but-3-yn-2-ol (16); 4-(4-(2-((5-Fluoropyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (17); 4-(4-(2-((4-Chloropyridin-2-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (18); 4-(4-(2-(Pyrimidin-4-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (19); 4-(4-(2-(Quinolin-2-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (20); 4-(4-(2-(Phenylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (21); 4-(4-(2-((3-Methoxyphenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (22); 3-((4-(2-(3-Hydroxy-3-(thiazol-2-yl)but-1-yn-1-yl)-6,7-dihydropyrazolo[1,5-a]pyrimidin-4(5H)-yl)pyrimidin-2-yl)amino)benzonitrile (23); 4-(4-(2-((3-(Dimethylamino)phenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (24); 4-(4-(2-((3-Morpholinophenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (25); 4-(4-(2-((3,5-Dimethoxyphenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (26); 4-(4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (27); 4-(4-(2-((2,2-Difluorobenzo[d][1,3]dioxol-5-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (28); 4-(4-(2-((4-Fluorophenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (29); 4-(4-(2-((4-Chlorophenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (30); 4-(4-(2-((1-Methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (31); 4-(4-(2-((1-Isopropyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (32); 4-(4-(2-((1-Methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (33); 4-(4-((4-Fluorophenyl)amino)-1,3,5-triazin-2-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (34); 4-(4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-methylbut-3-yn-2-ol (35); 3-((4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)ethynyl)oxetan-3-ol (36); 1-((4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)ethynyl)cyclopentan-1-ol (37); 7-((4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)ethynyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-7-ol (38); 4-(4-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)-2-(5-methylisoxazol-3-yl)but-3-yn-2-ol (39); 4-(7-(2-Aminopyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (40); 4-(7-(2-(Bicyclo 1.1.1]pentan-1-ylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (41); 4-(7-(2-((1-Methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (42); 4-(7-(2-((1-Methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (43); 4-(7-(2-((1-(Tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (44); 4-(7-(2-((1-(2-(Dimethylamino)ethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (45); 4-(7-(2-((1-Methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (46); 4-(7-(2-(Phenylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (47); 4-(7-(2-((3-Methoxyphenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (48); 3-((4-(2-(3-Hydroxy-3-(thiazol-2-yl)but-1-yn-1-yl)benzofuran-7-yl)pyrimidin-2-yl)amino)benzonitrile (49); 4-(7-(2-((3-(Dimethylamino)phenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (50); 4-(7-(2-((3-Morpholinophenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (51); 4-(7-(2-((4-Fluorophenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (52); 4-(7-(2-(((4-Chlorophenyl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (53); 4-(7-(2-(Benzo[d][1,3]dioxol-5-ylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (54); 4-(7-(2-((2,3-Dihydrobenzo[b][1,4]dioxan-6-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (55); 4-(7-(2-(Pyridin-2-ylamino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (56); 4-(7-(2-((6-(Dimethylamino)pyridin-2-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (57); 4-(7-(2-((6-Morpholinopyridin-2-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (58); 4-(7-(2-((6-Methoxypyridin-2-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (59); 4-(7-(2-((2-(Difluoromethyl)pyridin-4-yl)amino)pyrimidin-4-yl)benzofuran-2-yl)-2-(thiazol-2-yl)but-3-yn-2-ol (60) 5. The compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof, wherein, the pharmaceutically acceptable salt includes an acid addition salt formed by the compound of general formula (I) and the following acids, and the acids are hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, or an acid salt of an inorganic base containing an alkaline metal cation, an alkaline earth metal cation or an ammonium cation salt.

6. Use of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof in the preparation of a NIK inhibitor.

7. Use of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing or treating NIK target-mediated dependent diseases.

8. According to the use of claim 7, wherein, the NIK target-mediated dependent diseases are rheumatoid arthritis, gouty arthritis, systemic lupus erythematosus, Crohn's disease, ulcerative colitis, inflammatory bowel disease, psoriatic arthritis, psoriasis, type I diabetes, allergic dermatitis, myelofibrosis hyperplasia, polycythemia vera, acute toxic injury of liver and kidney, myeloma, sepsis or acute lymphocytic leukemia.

9. A pharmaceutical composition for preparing a drug for preventing or treating NIK-mediated diseases, wherein, the composition comprises the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable carrier or excipient.

10. According to the pharmaceutical composition of claim 9, wherein, the pharmaceutical composition is a capsule, powder, tablet, granule, pill, injection, syrup, oral preparation, inhalant, ointment, suppository or patch.

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