Macrocyclic compound, preparation method therefor and use thereof, and pharmaceutical composition

Abstract

A macrocyclic compound, a preparation method therefor and a use thereof, and a pharmaceutical composition. The compound has a structure represented by formula (I). The disclosed compound or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof exhibits excellent inhibitory activity against HPK1 kinase, and can also be used for preventing and / or treating HPK1-mediated diseases.

Description

A macrocyclic compound and a preparation method and application thereof, and a pharmaceutical composition TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, and particularly relates to a macrocyclic compound containing a pyrazole-substituted pyrimidine structure, a preparation method and application thereof, and a pharmaceutical composition. BACKGROUND

[0002] T cell receptor (TCR) mediated T cell activation plays a crucial role in thymic T cell development, T cell subset differentiation and effector T cell function. TCR can specifically recognize the antigen peptide presented by MHC (major histocompatibility complex) on the surface of antigen presenting cells, and convert the extracellular antigen peptide into a signal that can be transmitted to the cell interior through MHC recognition. Among them, the MHC molecules on the surface of antigen presenting cells include MHC class II and MHC class I molecules, which can be specifically recognized by CD4+ and CD8+ T cell surface corresponding helper receptors CD4, CD8 molecules, respectively, and then cause the activation of downstream signal pathways. The typical intracellular signals of TCR activation include mitogen-activated protein kinase (MAPK), protein kinase C (PKC) and calcium ion signal pathways. The activation of these signals finally activates the specific gene expression of T cells, causes the proliferation of cells, and makes T cells differentiate into effector T cells.

[0003] HPK1, also known as MAP4K1, is a serine / threonine kinase and is one of the members of the MAP4K family, which also includes five members, MAP3K2, MAP4K3, MAP4K4, MAP4K5 and MAP4K6. HPK1 can interact with many linker proteins, such as SLP-76 family, CARD11, HIS, HIP-55, GRB2 family, LAT, CRK family, etc., activate the JNK / SAPK signaling pathway of hematopoietic stem cells, and thus negatively regulate the TCR pathway. MAP4K3, also known as GLK kinase, has a biological effect opposite to that of HPK1. GLK can promote the activation of the TCR pathway by binding to downstream linker proteins. Therefore, we need to screen HPK1 inhibitors that are selective for GLK.

[0004] The main processes in which HPK1 is involved in regulating TCR are: (1) TCR binds with extracellular antigen through MHC to activate TCR pathway to transmit signals to downstream linker protein molecules; (2) the activated SLP76 of linker protein tyrosine kinase Lck and Zap70, and then phosphorylates HPK1; (3) the activated HPK1 phosphorylates the receptor protein SLP-76; (4) the phosphorylation of SLP-76 provides various protein binding sites for the 14-3-3 (TCR pathway inhibitor) receptor protein to form a complex; (5) the SLP-76 phosphorylated complex is involved in the down-regulation of the Erk signal pathway and the ubiquitination degradation process of SLP76, thereby causing the decrease of TCR signal pathway and T cell proliferation. In summary, HPK1 can negatively regulate the TCR signal pathway, and therefore, HPK1 can serve as a new regulatory mechanism of T cell-mediated immune response and become a new research hotspot of immune anti-tumor.

[0005] HPK1 inhibitors play an important role in malignant solid tumors or hematological cancers (such as acute myeloid leukemia, bladder epithelial cancer, breast cancer, colon cancer, lung cancer, pancreatic cancer, melanoma), autoimmune diseases (such as systemic lupus erythematosus, psoriatic arthritis) and inflammatory reactions.

[0006] At present, there is no drug on the market targeting HPK1, in order to better meet the clinical needs, we aim to develop a highly selective and active HPK1 inhibitor, and the present application provides a novel structure of selective HPK1 inhibitor with good physicochemical properties and drug properties.

[0007] SUMMARY

[0008] The purpose of the present application is to provide a macrocyclic compound containing a pyrazole-substituted pyrimidine structure.

[0009] In order to achieve the above purpose, the first aspect of the present application provides a compound or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the compound having a structure shown in formula (I):

[0010] In formula (I),

[0011] R4, R5, R6, R7 are each independently selected from at least one of H, hydroxyl, halogen, substituted or unsubstituted C 1-8 alkyl, substituted or unsubstituted C 1-6 alkoxy;

[0012] R2 or R3 are each independently selected from substituted or unsubstituted C 1-8 alkyl;

[0013] R1 is selected from phenyl, substituted or unsubstituted C 1-8substituted or unsubstituted C 2-8 substituted or unsubstituted C 1-8 substituted or unsubstituted C 3-8 substituted or unsubstituted C 2-8 substituted or unsubstituted C

[0014] substituted or unsubstituted C 1-6 substituted or unsubstituted C 2-4 substituted or unsubstituted C 3-6 substituted or unsubstituted C 3-8 substituted or unsubstituted C

[0015] The second aspect of the present application provides a method for preparing a compound having a structure shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the method comprising: contacting a compound shown in formula (A) with a compound shown in formula (B),

[0016] wherein, in formula (B), L is selected from a protecting group

[0017] In formula (A), R4, R5, R6, R7 have the same definitions as those in any one of claims 1-3.

[0018] Optionally, the method further comprises: deprotecting the intermediate obtained after the contacting reaction.

[0019] The third aspect of the present application provides use of the compound having a structure shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to the first aspect in the preparation of a medicament for preventing and / or treating a disease mediated by HPK1.

[0020] The fourth aspect of the present application provides a pharmaceutical composition containing an active ingredient and an excipient, wherein the active ingredient comprises at least one of the compound having a structure shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to the first aspect.

[0021] The compound having a structure shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof provided by the present application exhibits excellent inhibitory activity on HPK1 kinase, and can be used for preventing and / or treating a disease mediated by HPK1, exhibits good selectivity on GLK kinase, and exhibits good pharmacokinetic properties and in-vivo efficacy. DETAILED DESCRIPTION

[0022] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0023] The terminology used in this invention is explained as follows:

[0024] In this article, the wavy lines in each group Indicates the bonding location.

[0025] Halogens include fluorine, chlorine, bromine, and iodine.

[0026] "Morpholinyl" refers to The groups shown are wavy lines indicating the bonding positions with group R2; that is, the morpholino group is a substituent for group R2.

[0027] "Pyranyl" refers to The groups shown are wavy lines indicating the bonding positions with group R2; that is, the pyran group acts as a substituent for group R2.

[0028] “C 1-8 "Acetamino" refers to acetamino groups with a total number of 1-8 carbon atoms, including C. 1-8 linear acetylamino and C 1-8 The branched acetamino group can be a straight-chain acetamino group or a branched acetamino group with a total number of carbon atoms of 1, 2, 3, 4, 5, 6, 7 or 8, such as acetamino, methylacetamino, ethylacetamino, isopropylacetamino, n-butylacetamino, isobutylacetamino, N,N-dimethylacetamino, etc.

[0029] “(C 1-6 "(alkyl)2N-" indicates that the two H atoms on the amino group are replaced by branched or straight-chain alkyl groups with 1, 2, 3, 4, 5, or 6 carbon atoms, and the two C atoms are... 1-6 The alkyl groups can be the same or different; for example, they can be (CH3)2N-, (CH3CH2)2N-, etc. etc. "(C 1-3 The alkyl group (2N-) has a similar explanation, except that the number of carbon atoms is different.

[0030] “C 2-4 "Acyl group" refers to an acyl group with a total number of carbon atoms of 2-4, such as acetyl, CH3CH2CO-, etc.

[0031] “C 3-6"Cycloalkyl" refers to cycloalkyl groups with a total number of carbon atoms of 3-6. For example, cycloalkyl groups with 3, 4, 5 or 6 carbon atoms can be cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0032] "Spiroheterocyclic group" indicates that one ring originates from a specific cyclic carbon atom on another ring, the two ring systems share a single carbon atom, and at least one ring system contains one or more N, O, or S heteroatoms. Each ring system contains 3-8 membered rings, and the H atoms on each ring can be arbitrarily substituted or unsubstituted by the corresponding groups defined herein. For example, it can be... etc., where the wavy line represents the bonding position with the structure shown in equation (I).

[0033] "Bridged heterobicyclic group" refers to a saturated bridged ring system, involving a non-aromatic bicyclic system, wherein at least one ring system contains one or more N, O, S heteroatoms, wherein each ring system contains 3-8 membered rings, and the H on each ring can be arbitrarily substituted or unsubstituted by the corresponding groups defined herein. For example, it can be... etc., where the wavy line represents the bonding position with the structure shown in equation (I).

[0034] "C replaced or not replaced" 1-8 "alkyl" refers to an alkyl group with a total number of 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, including C14 and C24. 1-8 straight-chain alkyl and C 1-8 Branched alkyl groups, and C 1-8 The H atom on the alkyl group may be substituted or left unsubstituted by the corresponding group defined herein, and the carbon atom in the substituent is not counted in the C10. 1-8 The total number of carbon atoms in alkyl groups.

[0035] "C replaced or not replaced" 1-8 "Sulfonyl group" indicates a sulfonyl group with a total number of 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, including C. 1-8 straight-chain sulfonyl group and C 1-8 The branched sulfonyl group, and C 1-8 The H on the sulfonyl group can be arbitrarily replaced or left unsubstituted by the corresponding group defined herein, and the carbon atom in the substituent is not counted in the C10. 1-8 The total number of carbon atoms in the sulfonyl group. "Substituted or unsubstituted C" 1-6 The sulfonyl group has a similar explanation, except that the number of carbon atoms in the sulfonyl group is different.

[0036] "C replaced or not replaced" 3-8 "monocyclic sulfonyl" and "substituted or unsubstituted C" 3-6 The monocyclic sulfonyl group has a similar definition, C 3-8substituted or unsubstituted C 3-6 at least one H in the monocyclic sulfonyl group of the formula (C

[0037] "substituted or unsubstituted C 2-8 azacycloalkyl" means a cycloalkyl group having a total number of carbon atoms of 2 to 8, and at least one of the atoms forming the ring is N, and the H on the azacycloalkyl group can be optionally substituted with or without the corresponding substituents defined herein, the number of carbon atoms contained in the substituents not being included in the aforementioned total number of carbon atoms. For example, it can be and the like, wherein the wavy line indicates the bonding position to the structure shown in the formula (I). "substituted or unsubstituted C 3-7 azacycloalkyl" has a similar explanation as this, except that the number of carbon atoms is different.

[0038] "substituted or unsubstituted C 2-8 azacycloalkyl" and "substituted or unsubstituted C 2-6 azacycloalkyl" have a similar definition as this, C 2-8 azacycloalkyl" and "substituted or unsubstituted C 2-6 azacycloalkyl" have a similar definition as this, C 1-8 azacycloalkyl" have a similar explanation as this, except that the number of carbon atoms is different.

[0039] In the case of "substituted or unsubstituted" related groups of the present application, if it is a substituted group, the substituents can be at any position capable of being substituted in the corresponding group, unless otherwise specified.

[0040] As described above, the first aspect of the present application provides a compound or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the compound having a structure shown in the formula (I):

[0041] wherein, in the formula (I),

[0042] R4, R5, R6, R7 are each independently selected from at least one of H, hydroxy, halogen, substituted or unsubstituted C 1-8 alkyl, substituted or unsubstituted C 1-6 alkoxy;

[0043] R2or R3are each independently selected from substituted or unsubstituted C 1-8 alkyl;

[0044] R1is selected from phenyl, substituted or unsubstituted C 1-8 acetylamino, substituted or unsubstituted C 2-8 azacycloalkyl, spiro heterobicyclic group, bridged heterobicyclic group, substituted or unsubstituted C 1-8 sulfonyl, substituted or unsubstituted C 3-8Monocyclic sulfonyl, substituted or unsubstituted C 2-8 The nitrogen-containing heterocyclic sulfonyl group;

[0045] R1 may contain any of the following substituents: H, fluorine, hydroxyl, cyano, (C) 1-6 Alkyl)2N-, morpholino, pyrano, methyl, ethyl, n-propyl, isopropyl, isobutyl, (CH3)2OHCH2-, C 2-4 Acyl group, C 3-6 cycloalkyl, substituted or unsubstituted C 3-8 At least one of the nitrogen-containing heterocyclic alkyl groups.

[0046] Preferably, in formula (I),

[0047] R4, R5, R6, and R7 are each independently selected from at least one of H, hydroxyl, fluorine, methyl, and methoxy.

[0048] R2 or R3 are each independently selected from methyl groups;

[0049] R1 is selected from phenyl, NH2COCH2-, (CH3)2NCOCH2-, substituted or unsubstituted C. 3-7 Azahexaalkyl, spirohexabicycloyl, bridged hexabicycloyl, substituted or unsubstituted C 1-6 sulfonyl, substituted or unsubstituted C 3-6 Monocyclic sulfonyl, substituted or unsubstituted C 2-6 The nitrogen-containing heterocyclic sulfonyl group;

[0050] R1 may contain any of the following substituents: H, fluorine, hydroxyl, cyano, (CH3CH2)2N-, morpholino, pyrano, methyl, ethyl, n-propyl, isopropyl, isobutyl, (CH3)2OHCH2-, acetyl, cyclopropyl, cyclobutyl, cyclohexyl, or substituted or unsubstituted C. 3-6 At least one of the nitrogen-containing heterocyclic alkyl groups.

[0051] In a preferred embodiment, the compound is selected from any one of the following compounds:

[0052] As previously described, a second aspect of the present invention provides a method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the method comprising: reacting a compound of formula (A) with a compound of formula (B) in a contact reaction.

[0053] In equation (B), L is selected from the protecting group.

[0054] In formula (A), the definitions of R4, R5, R6 and R7 are the same as those in any one of claims 1-3.

[0055] Optionally, the method further comprises: subjecting the intermediate obtained after the contact reaction to a deprotection treatment.

[0056] It should be noted that, in the present application, when R1 does not contain active hydrogen, the structure containing L2 in formula (B) is used as a raw material to perform a contact reaction with the compound shown in formula (A); when R2 contains active hydrogen, the structure containing a protecting group L1 in formula (B) is used as a raw material to perform a contact reaction with the compound shown in formula (A). Meanwhile, the present application does not have a particular limitation on the protecting group, and a person skilled in the art can select as needed, for example, it can be tert-butyloxycarbonyl (Boc).

[0057] Preferably, the molar ratio of the compound shown in formula (A) to the compound shown in formula (B) is 1:1-2.

[0058] Preferably, the contact reaction is performed under the following conditions: temperature is 110-120℃, and time is 1-2h.

[0059] It should be noted that the present application does not have a particular limitation on the catalyst and the solvent used in the contact reaction, and a person skilled in the art can select as needed. For example, the compound shown in formula (A) is contacted with the compound shown in formula (B), anhydrous sodium carbonate, 1,4-dioxane and water in the presence of tetrakis-triphenylphosphine palladium.

[0060] The aforementioned preparation method of the present application can also involve various post-treatment operations known in the art, such as extraction, washing, filtration, drying, concentration under reduced pressure, column chromatography, etc., and the present application does not have a particular limitation thereon, and a person skilled in the art should not understand it as a limitation on the present application.

[0061] It should be noted that the raw materials involved in the preparation method of the present application can be synthesized according to the structure of the raw material, combined with the organic synthesis method in the art, or can be obtained by commercial purchase. The preparation methods of several raw materials are exemplarily given in the following of the present application, and a person skilled in the art should not understand it as a limitation on the present application.

[0062] The third aspect of the present application provides the use of the compound having the structure shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof of the first aspect in the preparation of a drug for preventing and / or treating a HPK1-mediated disease.

[0063] Preferably, the HPK1-mediated disease is at least one of cancer or non-cancer proliferative disease, inflammation, and immune disease.

[0064] Preferably, the immune disease is at least one of lupus erythematosus, glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease, and autoimmune diabetes.

[0065] Preferably, the cancer or non-cancer proliferative disease is at least one of lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, breast cancer, breast ductal carcinoma, head and neck cancer, endometrial cancer, corpus uteri cancer, rectal cancer, liver cancer, kidney cancer, pancreatic cancer, renal pelvis cancer, esophageal cancer, esophageal adenocarcinoma, glioma, prostate cancer, thyroid cancer, female genital system cancer, carcinoma in situ, lymphoma, neurofibromatosis, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, oral cavity cancer, pharynx cancer, multiple myeloma, leukemia, non-Hodgkin's lymphoma, large intestinal villous adenoma, melanoma, cytoma, sarcoma, and myelodysplastic syndrome.

[0066] A fourth aspect of the present application provides a pharmaceutical composition containing an active ingredient and an excipient, wherein the active ingredient includes at least one of the compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to the first aspect.

[0067] Preferably, the content of the active ingredient is 1-99.9% by weight.

[0068] More preferably, in the antitumor drug, the content of the active ingredient is 5-99.9% by weight, and exemplarily, the content of the active ingredient is 10% by weight, 15% by weight, 20% by weight, 25% by weight, 30% by weight, 35% by weight, 40% by weight, 45% by weight, 50% by weight, 55% by weight, 60% by weight, 65% by weight, 70% by weight, 75% by weight, 80% by weight, 85% by weight, 90% by weight, etc.

[0069] In the present application, the excipient is various excipients conventionally used in the art, such as a solvent, an antioxidant, a filler, etc.

[0070] The present application will be described in detail below by way of examples. In the following examples, the raw materials used are commercially available analytical pure products without special instructions.

[0071] Example 1: Preparation of Compound 1

[0072] The synthetic route of Compound 1 is as follows:

[0073] In the synthetic route, -OTBS represents Boc represents tert-butyloxycarbonyl.

[0074] Step 1 : Preparation of tert-butyl (3-bromo-4-cyanophenyl)carbamate

[0075] Into a 500 mL single necked round bottom flask was added 4-amino-2-bromobenzonitrile (152.3 mmol) and N,N'-dimethylformamide (DMF, 250 mL) followed by N,N'- carbonyldiimidazole (CDI, 456.9 mmol). The reaction mixture was heated to 80 °C for 5 h, followed by the addition of tert-butanol (761.5 mmol) and continued heating at 80 °C for 2 h. TLC monitoring indicated complete consumption of starting material. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (1000 mL) and washed with saturated brine (500 mL) three times. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography to yield a white solid with 85.2% yield.

[0076] Step 2: Preparation of tert-butyl dimethyl(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)but-3-en-l-yloxy)silane

[0077] Into a 100 mL two necked round bottom flask was added bis(cyclopentadienyl)zirconium chloride (Schwartz reagent, 54.3 mmol) and purged with argon three times. This was followed by the sequential addition of 4-(tert-butyldimethylsilyloxy)-l-butyne (54.3 mmol), triethylamine (5.43 mmol) and finally the slow addition of pinacolborane (59.8 mmol) to the mixture. The resulting solution was heated to 60 °C and stirred under argon for 8 h. TLC monitoring indicated complete consumption of starting material. The mixture was cooled to room temperature and loaded onto a short silica gel column. Column chromatography using 10% ethyl acetate in hexanes yielded a colorless oil with 80.5% yield.

[0078] Step 3: Preparation of tert-butyl (E)-(3-(4-((tert-butyldimethylsilyl)oxy)but-l-en-l-yl)-4- cyanophenyl)carbamate

[0079] Into a 100 mL Schlenk tube was added tert-butyl (3-bromo-4-cyanophenyl)carbamate (16.8 mmol, palladium acetate 1.6 mmol), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (S-Phos, 3.36 mmol), cesium carbonate (50.4 mmol), and the resulting solution was purged with argon three times. Then tetrahydrofuran (THF, 20 mL), water (4.0 mL), (E)-tert-butyldimethyl(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)but-3-en-1- yloxy)silane (16.8 mmol) were added in sequence, and the resulting solution was heated to 80 °C and stirred under argon for 2 h. TLC monitoring showed that the starting material was completely consumed. The reaction mixture was transferred to a separatory funnel with ethyl acetate (500 mL), washed with saturated brine (200 mL) three times, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to give a light yellow oil in 79.4% yield.

[0080] Step 4: Preparation of tert-butyl (E)-(4-cyano-3-(4-hydroxybut-1-en-1-yl)phenyl)carbamate

[0081] Into a 250 mL single-necked round-bottom flask was added tert-butyl (E)-(3-(4-((tert- butyldimethylsilyl)oxy)but-1-en-1-yl)-4-cyanophenyl)carbamate (13.4 mmol), tetrahydrofuran (THF, 100 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1 mol / L, 20.1 mL). The resulting solution was stirred at room temperature for 8 h, and TLC monitoring showed that the starting material was completely consumed. The reaction mixture was transferred to a separatory funnel with ethyl acetate (500 mL), washed with saturated brine (200 mL) three times, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to give a colorless oil in 80.2% yield.

[0082] Step 5: Preparation of tert-butyl (4-cyano-3-(4-hydroxybutyl)phenyl)carbamate

[0083] Into a 250 mL single-necked round-bottom flask was added tert-butyl (E)-(4-cyano-3-(4- hydroxybut-1-en-1-yl)phenyl)carbamate (10.7 mmol), anhydrous methanol (100 mL), and 10% wt palladium-carbon hydrogenation catalyst (310 mg). The resulting solution was stirred at room temperature for 12 h, and then filtered under reduced pressure with diatomite filter aid. The filter cake was rinsed with anhydrous methanol, and the filtrate was concentrated under reduced pressure to give a white solid in 96.6% yield.

[0084] Step 6: Preparation of tert-butyl (S)-(2-hydroxy-1-(2-hydroxyphenyl)ethyl)carbamate

[0085] Into a 500 mL single necked round bottom flask was added (S)-2-(1-amino-2- hydroxyethyl)phenol (150.0 mmol), anhydrous ethanol (250 mL), di-tert-butyl dicarbonate (150.0 mmol). The reaction was stirred at room temperature for 1 h. The reaction was directly concentrated under reduced pressure to get a white solid. The yield was 97.6%.

[0086] Step 7: Preparation of (S)-(2-((tert-butyldimethylsilyl)oxy)-1-(2- hydroxyphenyl)ethyl)carbamic acid tert-butyl ester

[0087] Into a 500 mL single necked round bottom flask was added (S)-(2-hydroxy-1-(2- hydroxyphenyl)ethyl)carbamic acid tert-butyl ester (150.0 mmol), dichloromethane (DCM, 250 mL), imidazole (225.0 mmol), tert-butyldimethylsilyl chloride (165.0 mmol). The reaction was stirred at room temperature for 30 min. The reaction was directly concentrated under reduced pressure to get a colorless oil. The yield was 93.0%.

[0088] Step 8: Preparation of (S)-(1-(2-(4-(5-((tert-butoxycarbonyl)amino)-2- isocy anidophenyl)butyloxy)phenyl)-2-((tert-butyldimethylsilyl)oxy)ethyl)carbamic acid tert-butyl ester

[0089] Into a 100 mL single necked round bottom flask was added tert-butyl (4-cyano-3-(4- hydroxybutyl)phenyl)carbamate (10.3 mmol), triphenylphosphine (20.6 mmol), (S)-(2- ((tert-butyldimethylsilyl)oxy)-1-(2-hydroxyphenyl)ethyl)carbamic acid tert-butyl ester (10.3 mmol), toluene 10 mL. The reaction was stirred at 80 °C until the starting material was dissolved. Then diisopropyl azodicarboxylate (20.6 mmol) was added dropwise. The reaction was stirred for 30 min. The reaction was directly concentrated under reduced pressure to get a colorless oil. The yield was 50.1%.

[0090] Step 9: Preparation of (S)-4-amino-2-(4-(2-(1-amino-2-hydroxyethyl)phenoxy)butyl)benzonitrile hydrochloride

[0091] Into a 100 mL single necked round bottom flask was added (S)-(1-(2-(4-(5-((tert- butoxycarbonyl)amino)-2-oxazolyl)phenoxy)ethyl)pyrrolidin-2-yl)methanol (0.5 mmol), acetonitrile (5.0 mL), hydrogen chloride ethyl acetate solution (4 mol / L, 10.0 mL), the reaction was stirred at room temperature for 20 h, TLC monitoring of the reaction was complete, the reaction was filtered under reduced pressure, the filter cake was washed with anhydrous ethyl ether and dried to obtain a white solid, the yield was 88.4%.

[0092] Step 10: Preparation of (S)-4-amino-2-(4-(2-(1-((5-bromo-2-chloropyrimidin-4- yl)amino)-2-hydroxyethyl)phenoxy)butyl)benzonitrile

[0093] Into a 50 mL single necked round bottom flask was added (S)-4-amino-2-(4-(2-(1- amino-2-hydroxyethyl)phenoxy)butyl)benzonitrile hydrochloride (4.6 mmol), acetonitrile (5.0 mL), N,N'-diisopropylethylamine (DIPEA, 14.8 mmol), 5-bromo-2,4- dichloropyrimidine (4.6 mmol), after the addition was completed, the reaction was stirred at room temperature for 5 h, TLC monitoring of the reaction was complete, the reaction system was transferred to a separatory funnel with ethyl acetate (200 mL), washed with saturated brine (100 mL) for 3 times, dried with anhydrous sodium sulfate, concentrated under reduced pressure, the residue was purified by column chromatography to obtain a light yellow oil, the yield was 48.4%.

[0094] Step 11: Preparation of (S)-3 5 - bromo-5-hydroxymethyl-7-oxa-2,4-diazabicyclo[3.2.1]oct-6-ene-1,2-dicarboxamide 4 - methylcarbamide

[0095] Into a 50 mL single necked round bottom flask was added acetonitrile (10 mL), water (1.0 mL), hydrogen chloride dioxane solution (4 mol / L, 0.1 mL), heated to reflux, then (S)-4-amino-2-(4-(2-(1-((5-bromo-2-chloropyrimidin-4-yl)amino)-2- hydroxyethyl)phenoxy)butyl)benzonitrile (1.0 mmol) was dissolved in 3 mL of acetonitrile and added dropwise into the reaction flask, after the addition was completed, refluxed for 5 h, TLC monitoring of the reaction was complete, the system was directly concentrated under reduced pressure to obtain a yellow solid crude product, the crude product was purified by anhydrous methanol to obtain a white solid pure product, the yield was 30.4%.

[0096] Step 12: Preparation of Compound 1

[0097] Into a 10 mL microwave tube was added (S)-3 5- Bromo-5-hydroxymethyl-7-oxa-2,4-diazepin-3(2,4)-pyrimidin-1(1,3),6(1,2)- benzododecyl-1 4 - Cyanide (2.3 mmol), 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H- pyrazol-1-yl)acetonitrile (3.5 mmol), tetrakis triphenylphosphine palladium (0.2 mmol), anhydrous sodium carbonate (6.9 mmol), 1,4-dioxane (7.0 mL), water (1.5 mL), 115 °C for 1.5 h. TLC monitoring of the complete reaction of the starting material. The reaction system was transferred to a separatory funnel with ethyl acetate (150 mL), washed with saturated brine (100 mL) 3 times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography to obtain a light yellow solid with a yield of 30.8%.

[0098] 1 H NMR (600 MHz, DMSO-d6) δ 9.79 (s, 1H), 8.68 (s, 1H), 8.47 (m, 1H), 8.16 (s, 1H), 8.03 (s, 1H), 7.64-7.36 (m, 4H), 7.20 (m, 1H), 7.12-7.06 (m, 1H), 6.86 (m, 1H), 5.05 (s, 1H), 5.03-4.94 (m, 3H), 4.01-3.92 (m, 3H), 2.81-2.70 (m, 4H), 1.87-1.81 (m, 2H).

[0099] Example 2: Preparation of compound 2

[0100] The preparation method is similar to Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced with an equimolar amount of 1-phenylpyrazole-4-boronic acid pinacol ester to obtain compound 2 as a white solid with a yield of 58.5%.

[0101] 1H NMR (600 MHz, DMSO-d6) δ 9.78 (s, 1H), 8.68 (s, 1H), 8.60 (s, 1H), 8.47 (m, 2H), 7.83-7.78 (m, 2H), 7.64 (d, J = 8.5 Hz, 1H), 7.51 (m, 1H), 7.50-7.43 (m, 3H), 7.39-7.31 (m, 2H), 7.20 (m, 1H), 7.06 (m, 1H), 6.84 (m, 1H), 5.11 (m, 1H), 4.99-4.94 (m, 1H), 4.01-3.92 (m, 3H), 2.81-2.70 (m, 2H), 1.88-1.74 (m, 4H).

[0102] Example 3: Preparation of compound 3

[0103] The preparation method is similar to Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced with equimolar amount of 1-acetamido-4-boronic acid pinacol ester, to obtain compound 3 as a white solid with a yield of 55.9%.

[0104] 1 H NMR (600 MHz, DMSO-d6) δ 9.76-8.68 (s, 2H), 8.47 (m, 1H), 8.16-8.03 (s, 4H), 7.64-7.36 (m, 4H), 7.20 (m, 1H), 7.12-7.06 (m, 1H), 6.86 (m, 1H), 5.08 (m, 1H), 4.85-4.74 (m, 1H), 4.06-3.88 (m, 5H), 2.80-2.70 (m, 2H), 1.88-1.74 (m, 4H).

[0105] Example 4: Preparation of compound 4

[0106] The preparation method is similar to Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced with equimolar amount of N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetamide, to obtain compound 4 as a white solid with a yield of 43.6%.

[0107] 1H NMR (600 MHz, DMSO-d6) δ 9.79 (s, 1H), 8.68 (s, 1H), 8.47 (m, 1H), 8.16 (s, 1H), 8.03 (s, 1H), 7.64-7.36 (m, 4H), 7.20 (m, 1H), 7.12-7.06 (m, 1H), 6.86 (m, 1H), 5.06 (m, 1H), 4.96-4.83 (m, 4H), 4.06-3.88 (m, 5H), 2.80-2.70 (m, 2H), 1.88-1.74 (m, 7H).

[0108] Example 5: Preparation of compound 5

[0109] The preparation method is similar to Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced with equimolar amount of 1-morpholine-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethanone to obtain compound 6 as a white solid with a yield of 63.5%.

[0110] 1 H NMR (600 MHz, DMSO-d6) δ 9.79 (s, 1H), 8.68 (s, 1H), 8.47 (m, 1H), 8.16 (s, 1H), 8.03 (s, 1H), 7.64-7.36 (m, 4H), 7.20 (m, 1H), 7.12-7.06 (m, 1H), 6.86 (m, 1H), 5.06 (m, 1H), 4.96-4.83 (m, 4H), 4.06-3.88 (m, 5H), 2.80-2.70 (m, 2H), 1.88-1.74 (m, 7H).

[0111] Example 6: Preparation of compound 6

[0112] The preparation method is similar to Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced with equimolar amount of 1-(piperazin-1-yl)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethanone hydrochloride to obtain compound 6 as a white solid with a yield of 56.4%.

[0113] 1 H NMR (600 MHz, DMSO-d6) δ 9.55 (s, 1H), 8.60 (s, 1H), 8.56 (m, 1H), 8.05 (s, 1H), 7.89 (s, 1H), 7.74-7.54 (m, 2H), 7.38-7.20 (m, 3H), 7.12-7.06 (m, 1H), 6.93 (m, 1H), 5.04 (m, 1H), 4.99-3.93 (m, 5H), 3.50-3.46 (m, 4H), 2.80 (m, 4H), 2.77-2.70 (m, 2H), 2.41 (m, 2H), 1.87-1.79 (m, 2H), 1.79-1.74 (m, 2H).

[0114] Example 7: Preparation of compound 7

[0115] The preparation method is similar to Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced with equimolar amount of 1-(azetidin-3-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole hydrochloride to obtain compound 7 as a white solid with a yield of 52.5%.

[0116] 1 H NMR (600 MHz, DMSO-d6) δ 9.55 (s, 1H), 8.60 (s, 1H), 8.56 (m, 1H), 8.05 (s, 1H), 7.89 (s, 1H), 7.74-7.54 (m, 2H), 7.38-7.20 (m, 3H), 7.12-7.06 (m, 1H), 6.93 (m, 1H), 5.04 (m, 1H), 4.99-3.93 (m, 5H), 3.50-3.46 (m, 4H), 2.80 (m, 4H), 2.77-2.70 (m, 2H), 2.41 (m, 2H), 1.87-1.79 (m, 2H), 1.79-1.74 (m, 2H).

[0117] Example 8: Preparation of compound 8

[0118] The preparation method is similar to that of Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced by equimolar amount of 1-(1-(methylsulfonyl)azetidin-3-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole to obtain compound 8 as a white solid with a yield of 47.3%.

[0119] 1 H NMR (600 MHz, DMSO-d6) δ 9.56 (s, 1H), 8.62 (s, 1H), 8.47 (m, 1H), 8.14 (s, 1H), 7.94 (s, 1H), 7.64 (d, J = 8.5 Hz, 1H), 7.54 (m, 1H), 7.38 (m, 1H), 7.24 (m, 1H), 7.20 (m, 1H), 7.12-7.06 (m, 1H), 6.85-5.26 (m, 2H), 4.94-4.75 (m, 2H), 4.35-3.92 (m, 5H), 3.48 (d, 2H), 3.44 (s, 1H), 2.81-2.70 (m, 3H), 1.88-1.74 (m, 4H).

[0120] Example 9: Preparation of compound 9

[0121] The preparation method is similar to that of Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced by equimolar amount of 2-hydroxy-1-(3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)azetidin-1-yl)ethanone to obtain compound 9 as a white solid with a yield of 51.4%.

[0122] 1 H NMR (600 MHz, DMSO-d6) δ 9.56 (s, 1H), 8.62 (s, 1H), 8.47 (m, 1H), 8.14 (s, 1H), 7.94 (s, 1H), 7.64 (d, J = 8.5 Hz, 1H), 7.54 (m, 1H), 7.38 (m, 1H), 7.24 (m, 1H), 7.20 (m, 1H), 7.12-7.06 (m, 1H), 6.85-5.26 (m, 2H), 4.94-4.75 (m, 2H), 4.35-3.92 (m, 5H), 3.48 (d, 2H), 3.44 (s, 1H), 2.81-2.70 (m, 3H), 1.88-1.74 (m, 4H).

[0123] Example 10: Preparation of compound 10

[0124] The preparation method is similar to example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced with equimolar amount of 1-(1-(tetrahydro-2H-pyran-4-yl)azetidin-3-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole to obtain compound 10 as a white solid with a yield of 55.5%.

[0125] 1 H NMR (600 MHz, DMSO-d6) δ 9.74-8.75 (m, 2H), 8.47 (m, 1H), 8.11 (s, 1H), 7.96 (d, 1H), 7.64 (m, 1H), 7.50 (m, 1H), 7.36 (m, 1H), 7.24-7.20 (m, 2H), 7.12-7.06 (m, 1H), 6.85 (m, 1H), 5.12 (t, 1H), 4.99-4.94 (m, 1H), 4.65-4.61 (m, 1H), 4.01-3.93 (m, 5H), 3.66 (m, 2H), 3.50 (m, 2H), 3.11 (s, 1H), 2.87 (m, 1H), 2.81 (d, 2H), 2.78-2.70 (m, 2H), 1.86-1.64 (m, 8H).

[0126] Example 11: Preparation of compound 11

[0127] The synthetic route of compound 11 is as follows:

[0128] In the synthetic route, Boc represents tert-butyloxycarbonyl.

[0129] Step 1: Preparation of tert-butyl (S)-3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate

[0130] Into a 250 mL single-necked round-bottom flask, (R)-tert-butyl 3-hydroxypyrrolidine-1-carboxylate (22.0 mmol), triethylamine (26.0 mmol) and dichloromethane (DCM, 100 mL) were added and stirred in an ice bath, then methanesulfonyl chloride (22.4 mmol) was added, and after the addition was completed, the ice bath was removed and the reaction was allowed to proceed at room temperature for 2 h. TLC monitoring showed that the raw material was completely reacted, then saturated brine was added and washed 3 times, the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid with a yield of 75.8%.

[0131] Step 2: Preparation of (R)-tert-butyl 3-(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrazol-l-yl)pyrrolidine-l-carboxylate

[0132] Into a 250 mL single necked flask was placed (S)-tert-butyl 3-((methylsulfonyl)oxy)pyrrolidine-l- carboxylate (6.5 mmol), 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (7.2 mmol), cesium carbonate (9.8 mmol) and 100 mL of acetonitrile, then heated to 90 °C for 6 h. TLC monitoring of the starting material was complete, cooled to room temperature, added 200 mL of ethyl acetate, washed with saturated brine three times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to give a white solid with a yield of 85.5%.

[0133] Step 3: Preparation of (R)-tert-butyl 3-(4-((S)-l 4 -cyano-5-(hydroxymethyl)-7-oxa-2,4-diazabicyclo[3.2.0]heptane-3 5 -carboxylate

[0134] Into a 10 mL microwave tube was placed (S)-3 5 -bromo-5-hydroxymethyl-7-oxa-2,4-diazabicyclo[3.2.0]heptane-l 4 -carboxylate (2.3 mmol), (R)-tert-butyl 3-(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrazol-l-yl)pyrrolidine-l-carboxylate (3.5 mmol), tetrakis(triphenylphosphine)palladium (0.2 mmol), anhydrous sodium carbonate (6.9 mmol), 1,4-dioxane (7.0 mL), water (1.5 mL), and heated to 115 °C for 1.5 h. TLC monitoring of the starting material was complete. The reaction was transferred to a separatory funnel with ethyl acetate (150 mL), washed with saturated brine three times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography to give a light yellow solid with a yield of 27.8%.

[0135] Step 4: Preparation of Compound 11

[0136] Into a 50 mL single necked flask was placed (R)-tert-butyl 3-(4-((S)-l 4 -cyano-5-(hydroxymethyl)-7-oxa-2,4-diazabicyclo[3.2.0]heptane-3 51.5 mmol of tert-butyl 1-(1-yl)-1H-pyrazole-1-yl)pyrrolidine-1-carboxylate, ethyl acetate hydrochloride solution (4 M / 10 mL), and ethyl acetate (10 mL) were reacted at room temperature for 12 h. The reaction was monitored by TLC until it was complete. The solid was filtered to give a pale yellow solid with a yield of 85%.

[0137] 1 H NMR(600MHz,DMSO-d6)δ9.76-9.51(m,2H),8.36(s,1H),8.32(s,1H),8.04(s,1H),7.86(s,1H),7.72-7.68 (m,1H),7.27-7.24(m,2H),7.11(d,J=8.3Hz,1H),7.03(d,J=8.0Hz,1H),6.95(t,J=8.4Hz,1H),5.99-5.89 (m,1H),5.31-5.28(m,1H),4.27-4.12(m,2H),3.80-3.76(m,1H),3.70-3.67(m,3H),3.64-3.61(m, 2H),3.43-3.37(m,4H),2.84-2.67(m,2H),2.48-2.38(m,2H),2.14-1.96(m,2H),1.86-1.78(m,2H).

[0138] Example 12: Preparation of Compound 12

[0139] Compound 11 (0.8 mmol), acetic acid (1.2 mmol), DIPEA (3.2 mmol), and 10 mL of DMF were added to a 25 mL round-bottom flask. Then, 5 mL of a HATU (1.6 mmol) solution in DMF was slowly added dropwise. After the addition was complete, the reaction was carried out at room temperature for 2 h. The reaction mixture was monitored by TLC until the starting material was completely reacted. The reaction mixture was then transferred to a separatory funnel with 15 mL of ethyl acetate, washed three times with 50 mL of saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography to give a white solid in 50.8% yield.

[0140] 1H NMR (600 MHz, DMSO-d6) δ 9.73 (s, 1H), 8.54 (s, 1H), 8.17 (d, J = 20.0 Hz, 1H), 7.97 (s, 1H), 7.84 (d, J = 10.9 Hz, 1H), 7.54 (d, J = 8.3 Hz, 1H), 7.25-7.19 (m, 2H), 7.07 (d, J = 8.3 Hz, 1H), 7.03 (d, J = 8.0 Hz, 1H), 6.96-6.89 (m, 1H), 6.51-6.48 (m, 1H), 5.91-5.88 (m, 1H), 5.10 (d, J = 42.1 Hz, 1H), 4.95 (s, 1H), 4.27-4.14 (m, 2H), 4.03-3.85 (m, 1H), 3.83-3.61 (m, 4H), 3.57-3.45 (m, 1H), 2.75-2.59 (m, 2H), 2.38 (d, J = 7.2 Hz, 2H), 2.10-2.01 (m, 2H), 1.99 (s, 3H), 1.87-1.72 (m, 2H).

[0141] Example 13: Preparation of compound 13

[0142] The preparation method is similar to that of Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced with equimolar amount of 4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)piperidine hydrochloride to obtain compound 13 as a white solid with a yield of 53.2%.

[0143] 1 H NMR (600 MHz, DMSO-d6) δ 9.73 (s, 1H), 8.54 (s, 1H), 8.17 (d, J = 20.0 Hz, 1H), 7.97 (s, 1H), 7.84 (d, J = 10.9 Hz, 1H), 7.54 (d, J = 8.3 Hz, 1H), 7.25-7.19 (m, 2H), 7.07 (d, J = 8.3 Hz, 1H), 7.03 (d, J = 8.0 Hz, 1H), 6.96-6.89 (m, 1H), 6.51-6.48 (m, 1H), 5.91-5.88 (m, 1H), 5.10 (d, J = 42.1 Hz, 1H), 4.95 (s, 1H), 4.27-4.14 (m, 2H), 4.03-3.85 (m, 1H), 3.83-3.61 (m, 4H), 3.57-3.45 (m, 1H), 2.75-2.59 (m, 2H), 2.38 (d, J = 7.2 Hz, 2H), 2.10-2.01 (m, 2H), 1.99 (s, 3H), 1.87-1.72 (m, 2H).

[0144] Example 14: Preparation of compound 14

[0145] The preparation method is similar to that in Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced with equimolar amount of 1-methyl-4-[4-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]piperidine to obtain compound 14 in white solid with a yield of 56.7%.

[0146] 1 H NMR (600 MHz, DMSO-d6) δ 9.79 (s, 1H), 8.69 (s, 1H), 8.47 (m, 1H), 8.12 (m, 1H), 7.79 (d, 1H), 7.64 (m, 1H), 7.50 (m, 1H), 7.36 (m, 1H), 7.24-7.20 (m, 2H), 7.12-7.06 (m, 1H), 6.85 (m, 1H), 5.01 (t, J = 5.4 Hz, 1H), 4.99-4.94 (m, 1H), 4.44-4.39 (m, 1H), 4.01-3.92 (m, 5H), 2.80-2.70 (m, 2H), 2.67 (m, 2H), 2.55 (m, 2H), 2.02 (m 2H), 1.87-1.74 (m, 7H).

[0147] Example 15: Preparation of compound 15

[0148] The preparation method is similar to that in Example 12, except that compound 11 in Example 12 is replaced with equimolar amount of compound 13 to obtain compound 15 in white solid with a yield of 54.8%.

[0149] 1H NMR (600 MHz, DMSO-d6) δ 9.73 (s, 1H), 8.53 (s, 1H), 8.15 (s, 1H), 7.97 (s, 1H), 7.81 (s, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.23 (t, J = 7.8 Hz, 1H), 7.19 (d, J = 7.5 Hz, 1H), 7.07 (dd, J = 8.6, 2.2 Hz, 1H), 7.02 (d, J = 8.2 Hz, 1H), 6.92 (t, J = 7.5 Hz, 1H), 6.47 (d, J = 9.4 Hz, 1H), 5.91 - 5.87 (m, 1H), 4.99 - 4.96 (m, 2H), 4.50 (t, J = 13.6 Hz, 2H), 4.29 - 4.16 (m, 2H), 3.78 - 3.75 (m, 1H), 3.69 - 3.65 (m, 1H), 3.24 (t, J = 12.9 Hz, 1H), 2.76 (t, J = 12.8 Hz, 1H), 2.72 - 2.59 (m, 2H), 2.17 - 2.08 (m, 2H), 2.05 (s, 3H), 2.04 - 1.91 (m, 3H), 1.88 - 1.75 (m, 3H).

[0150] Example 16: Preparation of compound 16

[0151] The preparation method is similar to Example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in step 12) is replaced with equimolar amount of 1-phenyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole to obtain compound 16 as a white solid with a yield of 55.3%.

[0152] 1H NMR (600 MHz, DMSO-d6) δ 9.79 (s, 1H), 8.69 (s, 1H), 8.47 (m, 1H), 8.12 (d, J = 0.6 Hz, 1H), 7.79 (d, 1H), 7.64 (d, 1H), 7.50 (m, 1H), 7.36 (m, 1H), 7.24-7.20 (m, 2H), 7.12-7.06 (m, 1H), 6.75 (m, 1H), 5.24 (t, J = 5.4 Hz, 1H), 4.99-4.94 (m, 1H), 4.44-4.38 (m, 1H), 4.01-3.93 (m, 5H), 3.75 (m, 2H), 3.62 (m, 2H), 3.52-3.47 (m, 1H), 2.85 (s, 2H), 2.80-2.71 (m, 2H), 2.44-2.04 (m, 4H), 1.88-1.73 (m, 4H).

[0153] Example 17: Preparation of compound 17

[0154] The preparation method is similar to Example 11, except that (R)-3-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester in step 1 is replaced by equimolar amount of 6-hydroxy-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester, to obtain compound 17 as a light yellow solid with a yield of 59.0%.

[0155] 1 H NMR (600 MHz, DMSO-d6) δ 9.79-9.71 (m, 2H), 8.16 (s, 1H), 8.02 (s, 1H), 7.94 (s, 1H), 7.82 (s, 1H), 7.68 (s, 1H), 7.28-7.21 (m, 2H), 7.01 (d, J = 8.2 Hz, 1H), 6.93 (d, J = 7.0 Hz, 1H), 6.85 (t, J = 8.2 Hz, 1H), 5.99-5.88 (m, 1H), 4.96 (m, 1H), 4.26-4.22 (m, 1H), 3.85-3.79 (m, 2H), 3.70-3.66 (m, 4H), 3.64-3.60 (m, 2H), 2.84-2.67 (m, 2H), 2.48-2.38 (m, 2H), 2.32-2.26 (m, 4H), 2.14-1.96 (m, 2H), 1.85-1.76 (m, 2H).

[0156] Example 18: Preparation of compound 18

[0157] The preparation method is similar to that in Example 11, except that (R)-3-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester in step 1 is replaced by 3-hydroxy-8-azabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester in an equal molar amount, to obtain compound 18 in a yellowish solid form with a yield of 56.8%.

[0158] 1 H NMR (600 MHz, DMSO-d6) δ 9.72 (s, 1H), 8.51 (s, 1H), 8.15 (s, 1H), 7.95 (s, 1H), 7.80 (s, 1H), 7.55-7.52 (m, 1H), 7.22 (s, 1H), 7.16 (d, J = 7.4 Hz, 1H), 7.09-7.02 (m, 2H), 6.93 (s, 1H), 6.47-6.41 (m, 1H), 5.91-5.82 (m, 1H), 5.03 (s, 1H), 4.73 (t, 1H), 4.30-4.17 (m, 2H), 3.88 (s, 2H), 3.79-3.75 (m, 1H), 3.68-3.65 (m, 1H), 2.74-2.61 (m, 2H), 2.28-2.25 (m, 2H), 2.15-2.01 (m, 4H), 1.94-1.91 (m, 4H), 1.86-1.76 (m, 4H).

[0159] Example 19: Preparation of compound 19

[0160] The preparation method is similar to that in Example 12, except that compound 11 in Example 12 is replaced by an equal molar amount of compound 18, to obtain compound 19 in a white solid form with a yield of 55.4%.

[0161] 1H NMR (600 MHz, DMSO-d6) δ 9.72 (s, 1H), 8.52 (s, 1H), 8.14 (s, 1H), 7.95 (s, 1H), 7.78 (s, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.23 (t, J = 7.8 Hz, 1H), 7.17 (d, J = 7.5 Hz, 1H), 7.06 (d, J = 8.6 Hz, 1H), 7.03 (d, J = 8.2 Hz, 1H), 6.93 (t, J = 7.6 Hz, 1H), 6.43 (d, J = 9.4 Hz, 1H), 5.91-5.85 (m, 1H), 4.98 (t, 1H), 4.92-4.85 (m, 1H), 4.62-4.57 (m, 1H), 4.38-4.33 (m, 1H), 4.20 (d, J = 20.8, 11.2 Hz, 2H), 3.77-3.74 (m, 1H), 3.69-3.65 (m, 1H), 2.74-2.61 (m, 2H), 2.17-2.05 (m, 6H), 2.04 (s, 3H), 1.93-1.89 (m, 2H), 1.88-1.77 (m, 4H).

[0162] Example 20: Preparation of compound 20

[0163] Step 1: Preparation of 1-(methylsulfonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H- pyrazole

[0164] Into a 100 mL single necked round bottom flask was added 4-pyrazoleboronic acid pinacol ester (5.15 mmol), N,N-dimethylformamide (25.0 mL), placed in an ice bath with stirring, then added sodium hydride (10.3 mmol), stirred for five minutes then added dropwise methylsulfonyl chloride (10.3 mmol), after the addition was complete the ice bath was removed and the reaction was allowed to stir at room temperature for 12 h, TLC monitoring showed the reaction to be complete, the reaction was then poured into saturated ammonium chloride solution (200 mL), stirred until clear then the reaction was transferred to a separatory funnel, extracted with ethyl acetate three times, the organic phase was washed with saturated brine (100 mL) twice, dried over anhydrous sodium sulfate, concentrated under reduced pressure, the residue was purified by column chromatography to give a light yellow oil, the yield was 38.4%.

[0165] Step 2: Preparation of compound 20

[0166] Into a 10 mL microwave tube was added (S)-3 5 - bromo-5-hydroxymethyl-7-oxa-2,4-diazabicyclo[3.2.1]oct-6-yl 4- methanesulfonyl chloride (2.3 mmol), 1 -(methylsulfonyl)-4-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)-1 H-pyrazole (3.5 mmol), tetrakis(triphenylphosphine) palladium (0.23 mmol), anhydrous sodium carbonate (6.9 mmol), 1,4-dioxane (7.0 mL), water (1.5 mL), 115 °C for 1.5 h. TLC monitoring of the reaction until the starting material was consumed. The reaction mixture was transferred to a separatory funnel with ethyl acetate (150 mL), washed with saturated brine (100 mL) three times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography to give a white solid with a yield of 29.8%.

[0167] 1 H NMR (600 MHz, DMSO-d6) δ 9.83 (s, 1 H), 8.61 (s, 1 H), 8.54 (s, 1 H), 8.29 (s, 1 H), 8.03 (s, 1 H), 7.55 (d, J = 8.5 Hz, 1 H), 7.23 (s, 2H), 7.08 (d, J = 8.7 Hz, 1 H), 7.02 (d, J = 8.1 Hz, 1 H), 6.91 (s, 1 H), 6.66 (d, J = 9.5 Hz, 1 H), 5.91 (m, 1 H), 4.94 (m, 1 H), 4.23 (m, 1 H), 4.18 (m, 1 H), 3.95 (m, 1 H), 3.77 (m, 1 H), 3.60 (m, 1 H), 2.84 (s, 3H), 2.63 (m, 1 H), 2.04 (m, 2H), 1.81 (m, 2H).

[0168] Example 21 : Preparation of compound 21

[0169] The preparation method was similar to Example 20, except that the methylsulfonyl chloride in Example 20 was replaced with an equimolar amount of ethylsulfonyl chloride to give compound 21 as a white solid with a yield of 44.0%.

[0170] 1H NMR (600 MHz, DMSO-d6) δ 9.83 (s, 1H), 8.60 (d, J = 3.4 Hz, 1H), 8.54 (s, 1H), 8.29 (s, 1H), 8.04 (s, 1H), 7.66-7.58 (m, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.24 (s, 1H), 7.09 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 7.9 Hz, 1H), 6.91 (t, J = 7.4 Hz, 1H), 6.66 (d, J = 9.4 Hz, 1H), 5.93 (m, 1H), 4.96 (m, 1H), 4.28-4.15 (m, 2H), 3.95 (m, 1H), 3.74 (m, 2H), 3.62 (m, 1H), 2.76-2.59 (m, 2H), 2.11-1.99 (m, 2H), 1.88-1.75 (m, 2H), 1.04 (m, 3H).

[0171] Example 22: Preparation of compound 22

[0172] The preparation method is similar to that of Example 20, except that the methylsulfonyl chloride in Example 20 is replaced with an equimolar amount of difluoromethylsulfonyl chloride to obtain compound 22 as a white solid with a yield of 37.1%.

[0173] 1 H NMR (600 MHz, DMSO-d6) δ 9.83 (s, 1H), 8.60 (d, J = 3.4 Hz, 1H), 8.54 (s, 1H), 8.29 (s, 1H), 8.04 (s, 1H), 7.66-7.58 (m, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.24 (s, 1H), 7.09 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 7.9 Hz, 1H), 6.91 (t, J = 7.4 Hz, 1H), 6.66 (d, J = 9.4 Hz, 1H), 5.93 (m, 1H), 4.96 (m, 1H), 4.28-4.15 (m, 2H), 3.95 (m, 1H), 3.74 (m, 2H), 3.62 (m, 1H), 2.76-2.59 (m, 2H), 2.11-1.99 (m, 2H), 1.88-1.75 (m, 2H), 1.04 (m, 3H).

[0174] Example 23: Preparation of compound 23

[0175] The preparation method is similar to that of Example 20, except that the methylsulfonyl chloride in Example 20 is replaced with an equimolar amount of propylsulfonyl chloride to obtain compound 23 as a white solid with a yield of 36.7%.

[0176] 1 H NMR (600 MHz, DMSO-d6) δ 9.83 (s, 1H), 8.60 (d, J = 3.4 Hz, 1H), 8.54 (s, 1H), 8.29 (s, 1H), 8.04 (s, 1H), 7.66-7.58 (m, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.24 (s, 1H), 7.09 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 7.9 Hz, 1H), 6.91 (t, J = 7.4 Hz, 1H), 6.66 (d, J = 9.4 Hz, 1H), 5.93 (m, 1H), 4.96 (m, 1H), 4.28-4.15 (m, 2H), 3.95 (m, 1H), 3.74 (m, 2H), 3.62 (m, 1H), 2.76-2.59 (m, 2H), 2.11-1.99 (m, 2H), 1.88-1.75 (m, 2H), 1.64 (m, 2H), 0.95 (m, 3H).

[0177] Example 24: Preparation of compound 24

[0178] The preparation method is similar to that of Example 20, except that methylsulfonyl chloride in Example 20 is replaced with an equimolar amount of isopropylsulfonyl chloride to obtain compound 24 as a white solid with a yield of 39.8%.

[0179] 1 H NMR (600 MHz, DMSO-d6) δ 9.83 (s, 1H), 8.60 (d, J = 3.4 Hz, 1H), 8.54 (s, 1H), 8.29 (s, 1H), 8.04 (s, 1H), 7.66-7.58 (m, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.24 (s, 1H), 7.09 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 7.9 Hz, 1H), 6.91 (t, J = 7.4 Hz, 1H), 6.66 (d, J = 9.4 Hz, 1H), 5.93 (m, 1H), 4.96 (m, 1H), 4.28-4.15 (m, 2H), 3.95 (m, 1H), 3.74 (m, 2H), 3.62 (m, 1H), 2.76-2.59 (m, 2H), 2.11-1.99 (m, 2H), 1.88-1.75 (m, 2H), 1.64 (m, 2H), 0.95 (m, 3H).

[0180] Example 25: Preparation of compound 25

[0181] The preparation method is similar to that in Example 20, except that the methylsulfonyl chloride in Example 20 is replaced by an equal molar amount of cyclopropylsulfonyl chloride to obtain compound 25 as a white solid with a yield of 38.2%.

[0182] 1 H NMR (600 MHz, DMSO-d6) δ 9.82 (s, 1H), 8.60 (d, J = 5.3 Hz, 1H), 8.54 (s, 1H), 8.28 (d, J = 4.9 Hz, 1H), 8.05 (d, J = 4.6 Hz, 1H), 7.55 (d, J = 8.3 Hz, 1H), 7.29-7.21 (m, 2H), 7.09 (d, J = 8.2 Hz, 1H), 7.02 (d, J = 7.8 Hz, 1H), 6.92 (d, J = 9.6 Hz, 1H), 6.72 (s, 1H), 5.93 (m, 1H), 4.94 (m, 1H), 3.78 (m, 1H), 3.62 (m, 1H), 3.19 (m, 1H), 2.72 (m, 1H), 2.65 (m, 1H), 2.03 (m, 2H), 1.87-1.74 (m, 2H), 1.35 (m, 2H), 1.25 (m, 2H), 1.06 (m, 2H).

[0183] Example 26: Preparation of compound 26

[0184] The preparation method is similar to that in Example 20, except that the methylsulfonyl chloride in Example 20 is replaced by an equal molar amount of cyclopropylsulfonyl chloride to obtain compound 25 as a white solid with a yield of 38.2%.

[0185] 1 H NMR (600 MHz, DMSO-d6) δ 9.82 (s, 1H), 8.60 (d, J = 5.3 Hz, 1H), 8.54 (s, 1H), 8.28 (d, J = 4.9 Hz, 1H), 8.05 (d, J = 4.6 Hz, 1H), 7.55 (d, J = 8.3 Hz, 1H), 7.29-7.21 (m, 2H), 7.09 (d, J = 8.2 Hz, 1H), 7.02 (d, J = 7.8 Hz, 1H), 6.92 (d, J = 9.6 Hz, 1H), 6.72 (s, 1H), 5.93 (m, 1H), 4.94 (m, 1H), 3.78 (m, 1H), 3.62 (m, 1H), 3.19 (m, 1H), 2.72 (m, 1H), 2.65 (m, 1H), 2.03 (m, 2H), 1.87-1.74 (m, 2H), 1.35 (m, 2H), 1.25 (m, 2H), 1.06 (m, 2H).

[0186] Example 27: Preparation of compound 27

[0187] Step 1: Preparation of S-cyclobutyl thioacetate

[0188] Into a 250 mL single necked round bottom flask was added cyclobutyl bromide (30.2 mmol), potassium thioacetate (60.5 mmol), DMF (100 mL). The reaction was heated to 80 °C until TLC monitoring showed the starting material was consumed. The reaction was extracted with ethyl acetate (100 mL) three times, the organic phase was combined and washed with saturated brine (100 mL) twice, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography to give the product as a yellow oil in 64.8% yield.

[0189] Step 2: Preparation of cyclobutane sulfonyl chloride

[0190] Into a 250 mL single necked round bottom flask was added NCS (19.3 mmol), acetonitrile (100 mL), concentrated hydrochloric acid (5 mL) and stirred at room temperature for 15 minutes. A solution of S-cyclobutyl thioacetate (16.1 mmol) in acetonitrile (20 mL) was added under ice bath and the reaction was stirred at ice bath until TLC monitoring showed the starting material was consumed. The reaction was poured into saturated sodium bicarbonate solution (200 mL) and extracted with methyl tert-butyl ether (100 mL) three times, the organic phase was combined and washed with saturated brine (100 mL) twice, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product as a yellow oil which was used directly in the next step.

[0191] The subsequent preparation was similar to Example 20 except that methyl sulfonyl chloride in Example 20 was replaced with an equimolar amount of cyclobutyl sulfonyl chloride. White solid compound 27 was obtained in 34.6% yield.

[0192] 1 H NMR (600 MHz, DMSO-d6) δ 9.84 (s, 1H), 8.62 (s, 1H), 8.55 (s, 1H), 8.30 (s, 1H), 8.07 (s, 1H), 7.55 (s, 1H), 7.23 (s, 2H), 7.09 (s, 1H), 7.02 (s, 1H), 6.90 (s, 1H), 6.65 (s, 1H), 5.92 (m, 1H), 4.97 (m, 1H), 4.25 (m, 3H), 4.18 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H), 3.78 (m, 1H),, 2.71 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.79 (m, 2H), 1.59 (m, 1H), 1.13 (m, 2H).

[0193] Example 28: Preparation of compound 28

[0194] The preparation method is similar to example 20, except that methylsulfonyl chloride in example 20 is replaced by equimolar amount of cyclohexylsulfonyl chloride, to obtain compound 28 as a white solid with a yield of 40.5%.

[0195] 1 H NMR (600 MHz, DMSO-d6) δ 9.84 (s, 1H), 8.59 (s, 1H), 8.54 (s, 1H), 8.29 (s, 1H), 8.04 (s, 1H), 7.61 (d, J = 10.8 Hz, 1H), 7.26-7.18 (m, 2H), 7.09 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 8.0 Hz, 1H), 6.91 (d, J = 7.8 Hz, 1H), 6.62 (d, J = 9.5 Hz, 1H), 5.91 (m, 1H), 4.95 (m, 1H), 4.24 (m, 1H), 4.18 (m, 1H), 3.94 (m, 1H), 3.76 (m, 2H), 3.62 (m, 1H), 2.71 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.90 (m, 2H), 1.78 (m, 2H), 1.59 (m, 1H), 1.44 (m, 2H), 1.29 (m, 2H), 0.87-0.78 (m, 2H).

[0196] Example 29: Preparation of compound 29

[0197] The preparation method is similar to example 1, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)acetonitrile in example 1 is replaced by equimolar amount of 1-(phenylsulfonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole, to obtain compound 28 as a white solid with a yield of 40.5%.

[0198] 1H NMR (600 MHz, DMSO-d6) δ 9.82 (s, 1H), 8.80 (s, 1H), 8.53 (s, 1H), 8.21 (s, 1H), 8.08 (d, J = 7.9 Hz, 2H), 8.01 (s, 1H), 7.83 (t, J = 7.6 Hz, 1H), 7.71 (t, J = 7.7 Hz, 2H), 7.63 (s, 1H), 7.25-7.19 (m, 2H), 7.08 (d, J = 8.5 Hz, 1H), 7.01 (d, J = 8.1 Hz, 1H), 6.91 (t, J = 7.6 Hz, 1H), 6.73 (d, J = 9.3 Hz, 1H), 5.91 (m, 1H), 4.94 (m, 1H), 4.27-4.21 (m, 1H), 4.18 (m, 1H), 3.77 (m, 1H), 3.60 (m, 1H), 2.71 (m, 1H), 2.65 (m, 1H), 2.04 (m, 2H), 1.88-1.73 (m, 2H).

[0199] Example 30: Preparation of compound 30

[0200] The preparation method is similar to that of Example 20, except that methylsulfonyl chloride in Example 20 is replaced with equimolar amount of dimethylamine sulfonyl chloride to obtain compound 30 as a white solid with a yield of 36.6%.

[0201] 1 H NMR (600 MHz, DMSO-d6) δ 9.82 (s, 1H), 8.80 (s, 1H), 8.53 (s, 1H), 8.21 (s, 1H), 8.08 (d, J = 7.9 Hz, 2H), 8.01 (s, 1H), 7.83 (t, J = 7.6 Hz, 1H), 7.71 (t, J = 7.7 Hz, 2H), 7.63 (s, 1H), 7.25-7.19 (m, 2H), 7.08 (d, J = 8.5 Hz, 1H), 7.01 (d, J = 8.1 Hz, 1H), 6.91 (t, J = 7.6 Hz, 1H), 6.73 (d, J = 9.3 Hz, 1H), 5.91 (m, 1H), 4.94 (m, 1H), 4.27-4.21 (m, 1H), 4.18 (m, 1H), 3.77 (m, 1H), 3.60 (m, 1H), 2.71 (m, 1H), 2.65 (m, 1H), 2.04 (m, 2H), 1.88-1.73 (m, 2H).

[0202] Example 31: Preparation of compound 31

[0203] The preparation method is similar to that in Example 20, except that the methyl sulfonyl chloride in Example 20 is replaced by an equimolar amount of diethylamine sulfonyl chloride to obtain white solid compound 31 with a yield of 37.7%.

[0204] 1 H NMR (600 MHz, DMSO-d6) δ 9.82 (s, 1H), 8.55 (d, J = 14.4 Hz, 2H), 8.18 (s, 1H), 8.02 (s, 1H), 7.62 (s, 1H), 7.55 (d, J = 8.6 Hz, 2H), 7.08 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 8.1 Hz, 1H), 6.90 (t, J = 7.6 Hz, 1H), 6.59 (d, J = 9.3 Hz, 1H), 5.91 (m, 1H), 4.97 (m, 1H), 4.28-4.21 (m, 1H), 4.18 (m, 1H), 3.77 (m, 1H), 3.63 (m, 1H), 3.41 (m, 4H), 2.70 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.81 (m, 2H), 1.08 (m, 6H).

[0205] Example 32: Preparation of compound 32

[0206] Into a 250 mL single-necked round-bottom flask was added azetidine hydrochloride (10.7 mmol), DIPEA (21.4 mmol), dichloromethane (100 mL). After stirring at room temperature for 5 min and stirring at -20 °C for another 5 min, sulfonyl chloride (21.4 mmol) was added dropwise into the reaction system at -20 °C, and the reaction was allowed to proceed at room temperature for 10 h. The system was diluted with dichloromethane (100 ml), transferred to a separatory funnel, and washed with 1M HCl (100 ml) twice. The lower organic phase was retained, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a yellow oily product which was directly used in the next step.

[0207] The subsequent preparation method is similar to that in Example 20, except that the methyl sulfonyl chloride in Example 20 is replaced by an equimolar amount of azetidine-1-sulfonyl chloride, and finally yellow solid compound 32 is obtained with a yield of 30.6%.

[0208] 1H NMR (600 MHz, DMSO-d6) δ 9.84 (s, 1H), 8.62 (s, 1H), 8.55 (s, 1H), 8.30 (s, 1H), 8.07 (s, 1H), 7.55 (s, 1H), 7.23 (s, 2H), 7.09 (s, 1H), 7.02 (s, 1H), 6.90 (s, 1H), 6.65 (s, 1H), 5.92 (m, 1H), 4.97 (m, 1H), 4.25 (m, 3H), 4.18 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H), 3.78 (m, 1H), 3.61 (m, 1H), 3.42 (s, 1H), 2.71 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.79 (m, 2H).

[0209] Example 33: Preparation of compound 33

[0210] The preparation method is similar to that of Example 32, except that the azetidine hydrochloride in Example 32 is replaced by an equimolar amount of 3-hydroxyazetidine hydrochloride, and finally compound 32 in the form of a yellow solid is obtained with a yield of 35.5%.

[0211] 1 H NMR (600 MHz, DMSO-d6) δ 9.84 (s, 1H), 8.62 (s, 1H), 8.55 (s, 1H), 8.30 (s, 1H), 8.07 (s, 1H), 7.55 (s, 1H), 7.23 (s, 2H), 7.09 (s, 1H), 7.02 (s, 1H), 6.90 (s, 1H), 6.65 (s, 1H), 5.92 (m, 1H), 4.97 (m, 1H), 4.25 (m, 3H), 4.18 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H), 3.78 (m, 1H), 3.61 (m, 1H), 3.42 (s, 1H), 2.71 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.79 (m, 2H).

[0212] Example 34: Preparation of compound 34

[0213] The preparation method is similar to that of Example 32, except that the azetidine hydrochloride in Example 32 is replaced by an equimolar amount of 3-hydroxyazetidine hydrochloride, and finally compound 32 in the form of a yellow solid is obtained with a yield of 35.5%.

[0214] 1H NMR (600 MHz, DMSO-d6) δ 9.84 (s, 1H), 8.62 (s, 1H), 8.55 (s, 1H), 8.30 (s, 1H), 8.07 (s, 1H), 7.55 (s, 1H), 7.23 (s, 2H), 7.09 (s, 1H), 7.02 (s, 1H), 6.90 (s, 1H), 6.65 (s, 1H), 5.92 (m, 1H), 4.97 (m, 1H), 4.25 (m, 3H), 4.18 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H), 3.78 (m, 1H), 3.61 (m, 1H), 2.71 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.79 (m, 2H).

[0215] Example 35: Preparation of compound 35

[0216] The preparation method is similar to that of Example 32, except that the azetidine hydrochloride in Example 32 is replaced by equimolar amount of 3-fluoropyrrolidine hydrochloride, and finally yellow solid compound 35 is obtained with a yield of 37.7%.

[0217] 1 H NMR (600 MHz, DMSO-d6) δ 9.84 (s, 1H), 8.62 (s, 1H), 8.55 (s, 1H), 8.30 (s, 1H), 8.07 (s, 1H), 7.55 (s, 1H), 7.23 (s, 2H), 7.09 (s, 1H), 7.02 (s, 1H), 6.90 (s, 1H), 6.65 (s, 1H), 5.92 (m, 1H), 4.97 (m, 1H), 4.25 (m, 3H), 4.18 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H), 3.88 (m, 1H), 3.51 (m, 1H), 2.71 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.79 (m, 2H).

[0218] Example 36: Preparation of compound 36

[0219] The preparation method is similar to that of Example 32, except that the azetidine hydrochloride in Example 32 is replaced by equimolar amount of azetidine-3-one hydrochloride, and finally yellow solid compound 36 is obtained with a yield of 32.5%.

[0220] 1H NMR (600 MHz, DMSO-d6) δ 9.84 (s, 1H), 8.62 (s, 1H), 8.55 (s, 1H), 8.30 (s, 1H), 8.07 (s, 1H), 7.55 (s, 1H), 7.23 (s, 2H), 7.09 (s, 1H), 7.02 (s, 1H), 6.90 (s, 1H), 6.65 (s, 1H), 5.92 (m, 1H), 4.97 (m, 1H), 4.25 (m, 3H), 4.18 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H), 3.78 (m, 1H), 3.61 (m, 1H), 3.12 (s, 3H), 2.71 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.79 (m, 2H).

[0221] Example 37: Preparation of compound 37

[0222] The preparation method is similar to that of Example 32, except that the azetidine hydrochloride in Example 32 is replaced by an equimolar amount of 3-methoxyazetidine hydrochloride, and finally compound 37 in the form of a yellow solid is obtained with a yield of 40.8%.

[0223] 1 H NMR (600 MHz, DMSO-d6) δ 9.84 (s, 1H), 8.62 (s, 1H), 8.55 (s, 1H), 8.30 (s, 1H), 8.07 (s, 1H), 7.55 (s, 1H), 7.23 (s, 2H), 7.09 (s, 1H), 7.02 (s, 1H), 6.90 (s, 1H), 6.65 (s, 1H), 5.92 (m, 1H), 4.97 (m, 1H), 4.25 (m, 3H), 4.18 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H), 3.78 (m, 1H), 3.61 (m, 1H), 3.12 (s, 3H), 2.71 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.79 (m, 2H).

[0224] Example 38: Preparation of compound 38

[0225] The preparation method is similar to that of Example 20, except that the methane sulfonyl chloride in Example 20 is replaced by an equimolar amount of pyrrolidine-1-sulfonyl chloride, and finally compound 38 in the form of a white solid is obtained with a yield of 40.6%.

[0226] 1H NMR (600 MHz, DMSO-d6) δ 9.83 (s, 1H), 8.59 (s, 1H), 8.54 (s, 1H), 8.20 (s, 1H), 8.04 (s, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7.22 (d, J = 8.1 Hz, 2H), 7.08 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 8.0 Hz, 1H), 6.92 (d, J = 7.9 Hz, 1H), 6.62 (d, J = 9.2 Hz, 1H), 5.91 (m, 1H), 4.97 (m, 1H), 4.23 (m, 1H), 4.18 (m, 1H), 3.77 (m, 1H), 3.63 (m, 1H), 3.44 (m, 4H), 2.69 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.77 (m, 6H).

[0227] Example 39: Preparation of compound 39

[0228] The preparation method is similar to that of Example 20, except that the methanesulfonyl chloride in Example 20 is replaced with an equimolar amount of piperidin-1- sulfonyl chloride to obtain compound 39 as a white solid with a yield of 35.9%.

[0229] 1 H NMR (600 MHz, DMSO-d6) δ 9.83 (s, 1H), 8.59 (s, 1H), 8.54 (s, 1H), 8.20 (s, 1H), 8.04 (s, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7.22 (d, J = 8.1 Hz, 2H), 7.08 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 8.0 Hz, 1H), 6.92 (d, J = 7.9 Hz, 1H), 6.62 (d, J = 9.2 Hz, 1H), 5.91 (m, 1H), 4.97 (m, 1H), 4.23 (m, 1H), 4.18 (m, 1H), 3.77 (m, 1H), 3.63 (m, 1H), 3.44 (m, 4H), 2.69 (m, 1H), 2.64 (m, 1H), 2.04 (m, 2H), 1.77 (m, 6H).

[0230] Example 40: Preparation of compound 40

[0231] Steps 1 and 2 are similar to those of Example 20, except that the methanesulfonyl chloride in Example 20 is replaced with an equimolar amount of 4-(chlorosulfonyl)piperidine-1- carboxylate tert-butyl ester to obtain M1 as a white solid.

[0232] Step 3: Preparation of compound 40

[0233] Into a 50 mL single necked round bottom flask was added compound Ml, a certain amount of EA diluted HC1-dioxane solution. After stirring at room temperature for 12 h, TLC monitoring showed that the starting material was completely reacted, the system was diluted with ether, and then filtered under reduced pressure, and the white solid compound 40 was obtained, with a yield of 69.5%.

[0234] 1 H NMR (600 MHz, DMSO-d6) δ 9.83 (s, 1H), 8.56 (d, J = 18.2 Hz, 2H), 8.22 (s, 1H), 8.04 (s, 1H), 7.62 (t, J = 9.4 Hz, 3H), 7.09 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 8.0 Hz, 1H), 6.91 (t, J = 7.6 Hz, 1H), 6.61 (d, J = 9.3 Hz, 1H), 5.91 (m, 1H), 4.97 (m, 1H), 4.27-4.12 (m, 2H), 3.78 (m, 1H), 3.64 (m, 1H), 3.52 (s, 1H), 3.35 (m, 2H), 2.76-2.59 (m, 2H), 2.45 (m, 1H), 2.05 (m, 2H), 1.82 (m, 2H), 1.58-1.49 (m, 4H), 1.45 (m, 2H).

[0235] Example 41: Preparation of compound 41

[0236] The preparation method was similar to that of Example 20, except that the methylsulfonyl chloride in Example 20 was replaced with an equimolar amount of 4-methyl-1-piperazine sulfonyl chloride to obtain white solid compound 41, with a yield of 42.3%.

[0237] 1H NMR (600 MHz, DMSO-d6) δ 9.83 (s, 1H), 8.57 (s, 1H), 8.54 (s, 1H), 8.25 (s, 1H), 8.04 (s, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7.24 (d, J = 7.7 Hz, 2H), 7.08 (d, J = 8.3 Hz, 1H), 7.02 (d, J = 8.2 Hz, 1H), 6.91 (s, 1H), 6.67 (d, J = 9.5 Hz, 1H), 5.91 (m, 1H), 4.95 (m, 1H), 4.24 (m, 1H), 4.19 (m, 1H), 3.77 (m, 1H), 3.65 (m, 4H), 3.27 (m, 4H), 2.68 (m, 3H), 2.04 (m, 2H), 1.81 (m, 2H).

[0238] Example 42: Preparation of compound 42

[0239] The preparation method is similar to Example 20, except that the methylsulfonyl chloride in Example 20 is replaced with equimolar amount of morpholine-4-sulfonyl chloride to obtain compound 42 as a white solid with a yield of 38.5%.

[0240] 1 H NMR (600 MHz, DMSO-d6) δ 9.83 (s, 1H), 8.57 (s, 1H), 8.54 (s, 1H), 8.25 (s, 1H), 8.04 (s, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7.24 (d, J = 7.7 Hz, 2H), 7.08 (d, J = 8.3 Hz, 1H), 7.02 (d, J = 8.2 Hz, 1H), 6.91 (s, 1H), 6.67 (d, J = 9.5 Hz, 1H), 5.91 (m, 1H), 4.95 (m, 1H), 4.24 (m, 1H), 4.19 (m, 1H), 3.77 (m, 1H), 3.65 (m, 4H), 3.27 (m, 4H), 2.68 (m, 3H), 2.04 (m, 2H), 1.81 (m, 2H).

[0241] Example 43: Preparation of compound 43

[0242] The preparation method is similar to that in Example 1, except that 1-tert-butoxycarbonyl-pyrazole-4-boronic acid pinacol ester in step 12) is replaced with equimolar amount of 1,3-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole to obtain compound 43 as a white solid with a yield of 57.3%.

[0243] 1 H NMR (600 MHz, DMSO-d6) δ 9.69 (s, 1H), 8.52 (s, 1H), 7.81 (s, 1H), 7.75 (d, J = 1.6 Hz, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.22 (t, J = 8.0 Hz, 1H), 7.09 (dd, J = 21.5, 8.1 Hz, 2H), 7.01 (d, J = 8.2 Hz, 1H), 6.89 (t, J = 7.5 Hz, 1H), 6.24 (d, J = 9.5 Hz, 1H), 5.89-5.80 (m, 1H), 4.84 (t, 1H), 4.27-4.12 (m, 2H), 3.86 (s, 3H), 3.73-3.70 (m, 1H), 3.64-3.60 (m, 1H), 2.76-2.67 (m, 1H), 2.66-2.60 (m, 1H), 2.13 (s, 3H), 2.09-1.97 (m, 2H), 1.88-1.78 (m, 2H).

[0244] Example 44: Preparation of compound 44

[0245] The preparation method is similar to that in Example 1, except that 1-tert-butoxycarbonyl-pyrazole-4-boronic acid pinacol ester in step 12) is replaced with equimolar amount of 1,3-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole to obtain compound 43 as a white solid with a yield of 57.3%.

[0246] 1H NMR (600 MHz, DMSO-d6) δ 9.70 (s, 1H), 8.54 (s, 1H), 7.74 (s, 1H), 7.54-7.46 (m, 2H), 7.22 (t, J = 7.8 Hz, 1H), 7.12 (d, J = 7.6 Hz, 1H), 7.08 (d, J = 8.6 Hz, 1H), 7.01 (d, J = 8.2 Hz, 1H), 6.90 (t, J = 7.5 Hz, 1H), 6.27 (d, J = 9.6 Hz, 1H), 5.89-5.81 (m, 1H), 4.84 (t, J = 5.3 Hz, 1H), 4.27-4.14 (m, 2H), 3.83 (s, 3H), 3.74-3.68 (m, 1H), 3.64-3.56 (m, 1H), 2.78-2.59 (m, 2H), 2.23 (s, 3H), 2.13-2.00 (m, 2H), 1.89-1.70 (m, 2H).

[0247] Example 45: Preparation of compound 45

[0248] The preparation method is similar to Example 1, except that (S)-2-(1-amino-2- hydroxyethyl)phenol in step 6 is replaced by equimolar amount of (S)-2-(1-amino-2- hydroxyethyl)-5-fluorophenol; 1-tert-butoxycarbonyl-pyrazole-4-boronic acid pinacol ester in step 12 is replaced by equimolar amount of 2-methyl-1-(4-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-1H-pyrazol-1-yl)propan-2-ol. Compound 45 is obtained as a white solid with a yield of 57.4%.

[0249] 1 1H NMR (600 MHz, DMSO-d6) δ 9.76 (s, 1H), 8.53 (s, 1H), 8.02 (s, 1H), 7.96 (s, 1H), 7.76 (s, 1H), 7.54 (d, J = 8.5 Hz, 1H), 7.17 (t, J = 7.6 Hz, 1H), 7.07 (d, J = 8.6 Hz, 1H), 6.95 (d, J = 11.2 Hz, 1H), 6.73 (s, 1H), 6.42 (d, J = 9.3 Hz, 1H), 5.89-5.79 (m, 1H), 4.98 (t, J = 6.1 Hz, 1H), 4.78 (s, 1H), 4.30-4.16 (m, 2H), 4.11 (s, 2H), 3.76-3.68 (m, 1H), 3.65-3.57 (m, 1H), 2.79-2.58 (m, 2H), 2.12-1.96 (m, 2H), 1.86-1.72 (m, 2H), 1.13 (s, 6H).

[0250] Example 46: Preparation of compound 46

[0251] The preparation method is similar to example 1, except that (S)-2-(1-amino-2- hydroxyethyl)phenol in step 6 is replaced by equimolar amount of (S)-2-(1-amino-2- hydroxyethyl)-5-methylphenol; 1-tert-butoxycarbonyl-pyrazole-4-boronic acid pinacol ester in step 12 is replaced by equimolar amount of 2-methyl-1-(4-(4,4,5,5-tetramethyl- 1,3,2-dioxaborinan-2-yl)-1H-pyrazol-1-yl)propan-2-ol. Compound 46 is obtained as a white solid with a yield of 50.4%.

[0252] 1 H NMR (600 MHz, DMSO-d6) δ 9.75 (s, 1H), 8.55 (s, 1H), 8.02 (s, 1H), 7.96 (s, 1H), 7.75 (s, 1H), 7.54 (d, J = 8.5 Hz, 1H), 7.07 (d, J = 8.5 Hz, 1H), 7.03 (d, J = 7.6 Hz, 1H), 6.85 (s, 1H), 6.72 (d, J = 7.7 Hz, 1H), 6.37 (d, J = 9.3 Hz, 1H), 5.90-5.79 (m, 1H), 4.92 (t, J = 6.8 Hz, 1H), 4.79 (s, 1H), 4.26-4.20 (m, 1H), 4.19-4.14 (m, 1H), 4.12 (s, 2H), 3.79-3.68 (m, 1H), 3.66-3.56 (m, 1H), 2.75-2.63 (m, 2H), 2.27 (s, 3H), 2.08-1.97 (m, 2H), 1.87-1.74 (m, 2H), 1.13 (s, 6H).

[0253] Example 47: Preparation of compound 47

[0254] The preparation method is similar to example 1, except that (S)-2-(1-amino-2- hydroxyethyl)phenol in step 6 is replaced by equimolar amount of (S)-2-(1-amino-2- hydroxyethyl)-5-methoxyphenol; 1-tert-butoxycarbonyl-pyrazole-4-boronic acid pinacol ester in step 12 is replaced by equimolar amount of 2-methyl-1-(4-(4,4,5,5-tetramethyl- 1,3,2-dioxaborinan-2-yl)-1H-pyrazol-1-yl)propan-2-ol. Compound 47 is obtained as a white solid with a yield of 56.4%.

[0255] 1H NMR (600 MHz, DMSO-d6) δ 9.74 (s, 1H), 8.55 (s, 1H), 8.01 (s, 1H), 7.95 (s, 1H), 7.75 (s, 1H), 7.54 (d, J = 8.5 Hz, 1H), 7.06 (t, J = 9.5 Hz, 2H), 6.61-6.58 (m, 1H), 6.49 (d, J = 8.6 Hz, 1H), 6.35 (d, J = 9.6 Hz, 1H), 5.84-5.78 (m, 1H), 4.90 (t, J = 5.2 Hz, 1H), 4.77 (s, 1H), 4.23-4.16 (m, 2H), 4.12 (s, 2H), 3.73 (s, 3H), 3.71-3.68 (m, 1H), 3.61-3.55 (m, 1H), 2.71-2.64 (m, 2H), 2.09-1.98 (m, 2H), 1.87-1.76 (m, 2H), 1.13 (s, 6H).

[0256] Example 48: Preparation of compound 48

[0257] Into a 25 mL round-bottom flask was added compound 47 (1.0 mmol), DCM (10 mL), stirred at -40 °C, then added a solution of boron tribromide (1.2 mmol) in DCM (10 mL), and after the addition was complete, the reaction was allowed to warm to room temperature and stirred for 2 h. After the reaction was complete, as monitored by TLC, the reaction was quenched with water (20 mL), then 2 g of sodium carbonate was added, stirred for 1 h, and washed with saturated brine 3 times (50 mL each time). The organic phase was rotary evaporated to give compound 48 as a white solid in a yield of 75.5%.

[0258] 1 H NMR (600 MHz, DMSO-d6) δ 9.73 (s, 1H), 9.33 (s, 1H), 8.58 (s, 1H), 8.00 (s, 1H), 7.94 (s, 1H), 7.74 (s, 1H), 7.54 (d, J = 8.5 Hz, 1H), 7.07 (d, J = 8.5 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 6.43 (s, 1H), 6.30 (dd, J = 16.2, 8.9 Hz, 2H), 5.83-5.74 (m, 1H), 4.87 (t, J = 5.3 Hz, 1H), 4.77 (s, 1H), 4.19-4.14 (m, 1H), 4.11 (s, 2H), 4.10-4.07 (m, 1H), 3.72-3.66 (m, 1H), 3.58-3.52 (m, 1H), 2.76-2.60 (m, 2H), 2.09-1.96 (m, 2H), 1.87-1.74 (m, 2H), 1.13 (s, 6H).

[0259] Test Example 1: Experiment of kinase inhibitory activity of the compound of the present application

[0260] Materials and Methods:

[0261] Preparation of compound working solution: The test compound was diluted with dimethyl sulfoxide (DMSO), and 50 nL of the compound at different concentrations (500 μM, 166.67 μM, 55.56 μM, 18.52 μM, 6.173 μM, 2.058 μM, 0.686 μM, 0.229 μM, 0.076 μM, and 0.025 μM) in a 384-well LDV plate was transferred to an experimental plate by a liquid handler ECHO, and the final concentration of the compound in the experimental plate was 5000 nM, 1667 nM, 556 nM, 185.19 nM, 61.73 nM, 20.58 nM, 6.86 nM, 2.29 nM, 0.76 nM, and 0.25 nM.

[0262] Experimental procedure:

[0263] The experiment was performed in a 384-well plate (Corning, product number 4512), and the total reaction system in each well was 20 μL. Each well contained 1 x kinase buffer (1 mM MOPS (pH 7.2), 5 mM MgCl2, 1 mM EGTA, 0.4 mM EDTA, 2.5 mM β-Glycerol-Phosphate, 0.05 mM DTT), 2 μM MBP protein, and 40 μM ATP, 5 μL ADP-Glo™ reagent, 10 μL kinase detection reagent, and 5 nM HPK1 or GLK. 5 nM HPK1 or GLK, MBP protein, and ATP were reacted for 60 minutes, 5 μL of ADP-Glo™ reagent was added and reacted for 60 minutes, 10 μL of kinase detection reagent was added, and incubated at room temperature for 60 minutes. The signal was detected using a multifunctional microplate reader (EnVision).

[0264] Data processing: The luminescence signal value was converted to the inhibition rate.

[0265] Inhibition rate % = (maximum value - sample value) / (maximum value - minimum value) x 100;

[0266] The minimum value indicates the value measured in a well to which no enzyme but DMSO and substrate were added, and the maximum value indicates the value measured in a well to which DMSO, enzyme, and substrate were added.

[0267] IC 50Calculation of values: Curve fitting was performed by XLFit in Excel (version 5.3.1.3) to obtain the concentration of the test compound at which the enzyme activity was inhibited by 50%, i.e. IC50 50 .

[0268] Table-1 In vitro kinase inhibitory activity of example compounds

[0269] Test Example 2: In vivo pharmacokinetic study in mice

[0270] The compounds provided in the foregoing examples were administered to mice in polyethylene glycol 400 aqueous solution (70%). Blood samples of about 40 μL were collected at 5 min, 15 min, 30 min, 1, 2, 4, 8, 24 h after administration for the intravenous group and at 15 min, 30 min, 1, 2, 4, 8, 24 h after administration for the oral group into heparinized Eppendorf tubes, and stored on ice until centrifugation. The plasma was collected after centrifugation at 8000 rpm for 6 min, and transferred to 96-well plates and stored at -20°C until LC-MS / MS detection.

[0271] The pharmacokinetic parameters after administration to mice were calculated using the non-compartment model of the WinNonlin software.

[0272] Peak concentration C max : measured value;

[0273] Area under the concentration-time curve AUC0-t value: calculated using the trapezoidal rule; AUC 0-∞ = AUC 0-t + Ct / ke, Ct is the blood concentration at the last measurable time point, and ke is the elimination rate constant;

[0274] Elimination half-life t 1 / 2 = 0.693 / ke;

[0275] Time to peak T max : time required to reach the peak concentration after administration;

[0276] Mean residence time MRT = AUMC / AUC, AUMC refers to the area under the first moment plasma concentration-time curve;

[0277] Clearance CL: apparent volume of distribution of the drug cleared from the body per unit time;

[0278] Absolute bioavailability F = Dose iv *AUC 0-t，ig / Dose ig *AUC 0-t，iv x 100%.

[0279] Table-2 Pharmacokinetic properties of compound 25 in mice

[0280] Test Example 3: Anti-tumor activity of the compounds of the present application in a mouse subcutaneous tumor model

[0281] The anti-tumor activity of the compounds of the present application was evaluated in a standard murine model of implanted tumors. The mouse colon cancer cell line MC38 was cultured, collected and subcutaneously inoculated into the flank of 5-6 week old female mice (C57BL / 6, Shanghai Slac Laboratory Animal Co. Ltd.). When the tumor volume reached about 100 mm3, the animals were randomly divided into solvent control group (70% PEG-400 in water) and dosing groups (6 animals in each group). Subsequently, the animals were intraperitoneally injected or gavaged with the compounds of the examples or antibodies in combination with the compounds of the examples (corresponding doses, the compounds were dissolved in 70% PEG-400 in water, and the antibodies were dissolved in PBS solution), starting at any time from 0 to 7 days after tumor cell inoculation, and once or twice per day during the experiment. 3

[0282] The effect of the compounds of the examples on tumor growth was investigated, and the specific index was T / C% or tumor inhibition rate TGI (%).

[0283] The tumor diameter was measured twice a week using a vernier caliper, and the tumor volume (V) was calculated according to the following formula:

[0284] V = 1 / 2 x a x b 2 ; where a and b represent the length and width, respectively.

[0285] T / C (%) = (T - T0) / (C - C0) x 100; where T and C are the tumor volumes at the end of the experiment; T0 and C0 are the tumor volumes at the beginning of the experiment.

[0286] Tumor inhibition rate (TGI) (%) = 100 - T / C (%)

[0287] When the tumor regressed, tumor inhibition rate (TGI) (%) = 100 - (T - T0) / T0 x 100

[0288] If the tumor volume was reduced compared to the initial volume, i.e. T < T0 or C < C0, it was defined as partial tumor regression (PR) ; if the tumor completely disappeared, it was defined as complete tumor regression (CR).

[0289] The comparison between the tumor volumes of the two groups was performed using a two-tailed Student's t-test, and P < 0.05 was defined as statistically significant difference.

[0290] ​The compound having the structure shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof provided by the present application exhibits excellent inhibitory activity on HPK1 kinase, good selectivity on GLK kinase, and good pharmacokinetic properties and in vivo efficacy. More specifically, for example, compound 23 has excellent HPK1 kinase inhibitory activity (IC 50 less than 1 nM) and good GLK kinase selectivity (IC 50 more than 100 nM).

[0291] The above describes the preferred embodiments of the present application, but the present application is not limited thereto. Within the technical concept of the present application, various simple modifications can be made to the technical solutions of the present application, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be considered as the disclosed content of the present application, and all fall within the protection scope of the present application.

Claims

1. A compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, The compound has a structure shown in formula (I): wherein, in formula (I), R4, R5, R6, R7are each independently selected from at least one of H, hydroxyl, fluorine, methyl, methoxyl; R4, R5, R6, R7are each independently selected from at least one of H, hydroxyl, halogen, substituted or unsubstituted C 1-8 alkyl, substituted or unsubstituted C 1-6 alkoxy; R2and R3are each independently selected from the group consisting of substituted or unsubstituted C 1-8 alkyl; R1is selected from phenyl, substituted or unsubstituted C 1-8 acetylamino, substituted or unsubstituted C 2-8 azepinyl, spiroazabicycloalkyl, bridgedazabicycloalkyl, substituted or unsubstituted C 1-8 sulfonyl, substituted or unsubstituted C 3-8 monocyclic sulfonyl, substituted or unsubstituted C 2-8 azepinyl, spiroazabicycloalkyl, bridgedazabicycloalkyl, substituted or unsubstituted C The optional substituents contained in R1are at least one of H, fluorine, hydroxyl, cyano, (C 1-6 alkyl)2N-, morpholinyl, pyranyl, methyl, ethyl, n-propyl, i-propyl, i-butyl, (CH3)2OHCH2-, C 2-4 acyl, C 3-6 cycloalkyl, substituted or unsubstituted C 3-8 heterocycloalkyl, and the like.

2. The compound of claim 1, wherein, In formula (I), R4, R5, R6, R7are each independently selected from at least one of H, hydroxyl, fluorine, methyl, methoxyl; R2or R3are each independently selected from methyl; R1is selected from phenyl, NH2COCH2-, (CH3)2NCOCH2-, substituted or unsubstituted C 3-7 azacycloalkyl, spiroazabicycloalkyl, bridgedazabicycloalkyl, substituted or unsubstituted C 1-6 sulfonyl, substituted or unsubstituted C 3-6 monocyclic sulfonyl, substituted or unsubstituted C 2-6 azacycloalkyl, spiroazabicycloalkyl, bridgedazabicycloalkyl, substituted or unsubstituted C The optional substituents contained in R1are at least one of H, fluorine, hydroxyl, cyano, (CH3CH2)2N-, morpholinyl, pyranyl, methyl, ethyl, n-propyl, i-propyl, i-butyl, (CH3)2OHCH2-, acetyl, cyclopropyl, cyclobutyl, cyclohexyl, substituted or unsubstituted C 3-6 nitrogen heterocycloalkyl.

3. The compound according to any one of claims 1-2, wherein, The compound is selected from any one of the following compounds:

4. A process for preparing a compound of the structure shown in Formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, The method comprises: contacting a compound represented by formula (A) with a compound represented by formula (B), wherein, in formula (B), L is selected from a protecting group In formula (A), R4, R5, R6, R7are defined as the same as the definitions in any one of claims 1-3; Optionally, the method further comprises: subjecting the intermediate obtained after the contacting reaction to a deprotection treatment.

5. Use of a compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, as claimed in any one of claims 1-3, in the preparation of a medicament for preventing and / or treating a HPK1-mediated disease.

6. Use according to claim 4, wherein, The HPK1-mediated disease is at least one of a cancer or a non-cancer proliferative disease, an inflammation, and an immune disease; Preferably, the immune disease is at least one of lupus erythematosus, glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease, and autoimmune diabetes; Preferably, the cancer or non-cancer proliferative disease is at least one of lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, breast cancer, breast ductal carcinoma, head and neck cancer, endometrial cancer, corpus uteri cancer, rectal cancer, liver cancer, kidney cancer, pancreatic cancer, renal pelvis cancer, esophageal cancer, esophageal adenocarcinoma, glioma, prostate cancer, thyroid cancer, female reproductive system cancer, carcinoma in situ, lymphoma, neurofibromatosis, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, oral cavity cancer, pharynx cancer, multiple myeloma, leukemia, non-Hodgkin's lymphoma, large intestinal villous adenoma, melanoma, cytoma, sarcoma, myelodysplastic syndrome.

7. A pharmaceutical composition comprising an active ingredient and an excipient, wherein the active ingredient comprises at least one of a compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, as claimed in any one of claims 1-3. Preferably, the active ingredient is contained in an amount of 1-99.9% by weight.

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