Macrocyclic pyrazolopyrimidine compound and application thereof

Abstract

The invention relates to the field of biological medicine, in particular to a macrocyclic pyrazolopyrimidine compound which has better immunomodulatory activity and selectivity, and further provides application of the macrocyclic pyrazolopyrimidine compound to prevention or treatment of TLR7-related diseases, in particular to application of the macrocyclic pyrazolopyrimidine compound to prevention or treatment of TLR7-related diseases. The invention also discloses application of the compound as a vaccine adjuvant, a photodynamic therapeutic agent and a coupling drug.

Description

Technical Field

[0001] The present application relates to the field of biomedicine, and in particular to macrocyclic pyrazolopyrimidine compounds and uses thereof. Background Art

[0002] The immune system recognizes and responds to various pathogens, thereby protecting organisms from pathogens. Immune system dysfunction can lead to a variety of diseases including autoimmune diseases, inflammatory diseases and cancer. The main immune strategies of vertebrates are innate immunity and adaptive immunity, and the innate immune system is key to microbial recognition and clearance, inflammatory response, and cell death pathways. Targeting the proteins and signaling pathways of the innate immune system to develop disease treatment drugs provides a variety of possibilities for drug research and development.

[0003] Toll-like receptors (TLRs) are a family of proteins that play an important role in the immune system and are key receptors of the innate immune system. As a pattern recognition receptor (PRRs), TLRs can recognize damage-associated molecular patterns (DAMPs) and pathogen-associated molecular patterns (PAMPs), induce the activation of the NF-κB pathway and the secretion of cytokines and chemokines, thereby activating innate and adaptive immune responses. Currently, there are 13 members of the TLR family that have been discovered (TLR1-13), of which 10 are human TLRs (TLR1-10), mostly distributed in immune cells, and the binding ligands of different subtypes of TLRs are specific. Since TLRs are key molecules for immune triggering, TLR targets have important value in the fields of anti-infection, anti-inflammatory and anti-tumor.

[0004] The TLR7 gene is located at Xp22.3 on chromosome X of the genome. The TLR7 protein is known as a nucleic acid sensing molecule. It is located in the endosomal membrane and can recognize nucleotides and nucleosides related to intracellular pathogenic microorganisms. It is activated in the presence of viral single-stranded RNA (ssRNA). The TLR7 molecule has two binding sites, which bind to specific ligands respectively. The first binding site can bind to single nucleosides such as adenine nucleosides and guanine nucleosides, and can also bind to small molecule nucleoside analogs (such as TLR7 / 8 agonist R848). The second site binds to ssRNA, which can affect the affinity between the first binding site and the ligand. The binding of ligand to TLR7 activates the MyD88 pathway, induces cytokine secretion through the MyD88-NF-κB-dependent signal cascade, and thus activates the immune response.

[0005] Studies have shown that TLR7 has important applications in drug development for inflammatory diseases, viral infections, cancer, and autoimmune diseases. TLR7 agonists have multiple uses, including anti-tumor, anti-viral infection, and use as vaccine adjuvants. Currently, multiple agonists targeting TLR7 have entered clinical research, but the only small molecule compound on the market in the field of TLR7 agonists is imiquimod. The main difficulty in the clinical development of TLR7 agonists is the lack of significant therapeutic effects or immune-related adverse reactions. Therefore, the design of new, efficient, and selective TLR7 small molecule agonists has important clinical value. Summary of the invention

[0006] The present invention aims to provide a novel macrocyclic pyrazole TLR7 small molecule agonist with better immunomodulatory activity and target selectivity. The present invention also provides the use of the pyridone macrocyclic pyrazole TLR7 small molecule agonist for preventing or treating diseases related to TLR7, as well as the use as a vaccine adjuvant and photodynamic therapy agent, thereby providing the following inventions:

[0007] Compound

[0008] In one aspect, the present application provides a compound having a structure as shown in formula (I), a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope label or a metabolite thereof,

[0009]

[0010] in,

[0011] R 1 Selected from: hydrogen, halogen, hydroxyl, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl;

[0012] Q is selected from -O-, -S-, -NH-;

[0013] Ring A is selected from phenyl, naphthyl, 5-6 membered heteroaryl and bicyclic heteroaryl, wherein the phenyl, naphthyl, 5-6 membered heteroaryl and bicyclic heteroaryl are optionally substituted by 1, 2 or 3 groups selected from the following: hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, =O, -NR 3 R 4 、-CH2R a 、-C(=O)R b 、-OR c, C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -(C1-C6 alkyl)OH, -(C1-C6 alkyl)NH2, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -S(=O)(=O)NH(C1-C3 alkyl), -NR d S(=O)(=O)(C1-C6 alkyl), -P(=O)R e R f , sulfonic acid, -N=NPh, -Si(C1-C3 alkyl)3, -C(O-C1-C6 alkyl)=CH2; wherein R a , R b , R c , R d , R e , R f Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C(=O)(C1-C6 alkyl); wherein R 3 , R 4 are each independently selected from hydrogen, C 1-6 alkyl, C(═O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy;

[0014] Ring B is selected from 5-6 membered nitrogen-containing heteroaryl and bicyclic heteroaryl, and the 5-6 membered nitrogen-containing heteroaryl and bicyclic heteroaryl are optionally substituted by 1, 2 or 3 groups selected from the following: hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)R b’ 、-OR c’ , C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -(C1-C6 alkyl)OH, -(C1-C6 alkyl)NH2, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -S(=O)(=O)NH(C1-C3 alkyl), -NR d’S(=O)(=O)(C1-C6 alkyl), -P(=O)R e’ R f’ , sulfonic acid, -N=NPh, -Si(C1-C3 alkyl)3; wherein R a’ , R b’ , R c’ , R d’ , R e’ , R f’ Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C(=O)(C1-C6 alkyl); wherein R 3’ , R 4’ Each is independently selected from hydrogen, C1-C6 alkyl, C(═O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy;

[0015] L is selected from -(CH2) m -W-(CH2) n -, wherein W is selected from -O-, -NH-, -N(Boc)-, -NHC(=O)-, -CH2-, -CH(C1-C6 alkyl)-, -C(C1-C6 alkyl)(C1-C6 alkyl)-, -C(=O)-, -C(=O)NH-, -N(C1-C6 alkyl)-, -N(C1-C6 alkyl)C(=O)-, -C(=O)N(C1-C6 alkyl)-, -S-, -S(=O)-, -S(=O)(=O)-, -P -(=O)(C1-C6 alkyl)-, -P(=O)(C1-C6 alkoxy)-, -P(=O)(OH)-, -OC(=O)-, -C(=O)O-, -NHS(=O)(=O)-, -S(=O)(=O)NH-, -N(C1-C6 alkyl)S(=O)(=O)-, -S(=O)(=O)N(C1-C6 alkyl)-, m and n are each independently selected from an integer between 0 and 6 (e.g., 0, 1, 2, 3, 4, 5 or 6);

[0016] T is selected from -(CH-T 1 ) p -(Z) q -(CH-T 2 ) r-, wherein Z is selected from -O-, -NH-, -NHC(=O)-, -CH2-, -CH(C1-C6 alkyl)-, -C(C1-C6 alkyl)(C1-C6 alkyl)-, -C(=O)-, -C(=O)NH-, -N(C1-C6 alkyl)-, -N(C1-C6 alkyl)C(=O)-, -C(=O)N(C1-C6 alkyl)-, -S-, -S(=O)-, -S(=O)(=O)-, -P(=O)(C1-C6 alkyl)- wherein p, q, and r are each independently selected from an integer between 0 and 6 (e.g., 0, 1, 2, 3, 4, 5, or 6), and p and r cannot be 0 at the same time, and T 1 , T 2 Each is independently selected from: hydrogen, halogen, nitro, hydroxyl, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, mercapto, -S-(C1-C6 alkyl).

[0017] In certain embodiments, R 1 Selected from: hydrogen, halogen, C1-C6 alkyl, hydroxyl.

[0018] In certain embodiments, R 1 Selected from: hydrogen, halogen, hydroxyl.

[0019] In certain embodiments, R 1 Selected from hydrogen and halogen.

[0020] In certain embodiments, R 1 Selected from hydrogen.

[0021] In certain embodiments, Q is -O-.

[0022] In certain embodiments, Ring A is a 5-6 membered heteroaryl, optionally substituted with 1, 2 or 3 substituents.

[0023] In certain embodiments, Ring A is phenyl, optionally substituted with 1, 2 or 3 substituents.

[0024] In certain embodiments, the substituents of ring A are selected from the following groups: hydrogen, halogen, cyano, nitro, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, =O, -NR 3 R 4 、-CH2R a、-C(=O)R b 、-OR c , C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -S(=O)(=O)NH(C1-C3 alkyl), -NR d S(=O)(=O)(C1-C6 alkyl), -P(=O)R e R f , -C(O-C1-C6 alkyl)=CH2; wherein R a , R b , R c , R d , R e , R f Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, -C(=O)(C1-C6 alkyl); wherein R 3 , R 4 are each independently selected from hydrogen, C 1-6 Alkyl, -C(=O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy.

[0025] In certain embodiments, R a , R b , R c , R d , R e , R f Each is independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -N(C1-C6 alkyl)2; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0026] In certain embodiments, R 3 , R 4 are each independently selected from hydrogen, C 1-6 alkyl.

[0027] In certain embodiments, the 3-6 membered heterocyclyl is substituted by one or more groups selected from the group consisting of hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo-substituted C1-C6 alkyl, and C1-C6 alkoxy.

[0028] In certain embodiments, the substituents of ring A are selected from the following groups: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, =O, -CH2R a 、-C(=O)R b 、-OR c , C3-C6 cycloalkyl, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NR d S(=O)(=O)(C1-C6 alkyl), -P(=O)R e R f , -C(O-C1-C6 alkyl)=CH2; wherein R a , R b , R c , R d , R e , R f Each is independently selected from hydrogen, -OH, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -N(C1-C6 alkyl)2; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, hydroxyl, C1-C6 alkyl, halogenated C1-C6 alkyl;

[0029] In certain embodiments, R a , R b , R c , R d , R e , R f Each is independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -N(C1-C6 alkyl)2; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl;

[0030] In certain embodiments, ring A is selected from phenyl, which is optionally substituted with 1, 2 or 3 groups selected from halogen, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4, -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more C1-C6 alkyl groups.

[0031] In certain embodiments, Ring B is a bicyclic heteroaryl.

[0032] In certain embodiments, the ring B is a 5-6 membered monoheteroaryl group containing 1, 2 or 3 nitrogen atoms.

[0033] In certain embodiments, the 5-6 membered monoheteroaryl group containing 1, 2 or 3 nitrogen atoms is selected from the following heteroaromatic rings:

[0034] In certain embodiments, Ring B is Optionally, Ring B is substituted with 1, 2 or 3 substituents.

[0035] In certain embodiments, Ring B is Optionally, Ring B is substituted with 1, 2 or 3 substituents.

[0036] In certain embodiments, the substituents of ring B are selected from the following groups: hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo-C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)R b’ 、-OR c’ , C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -(C1-C6 alkyl)OH, -(C1-C6 alkyl)NH2, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -S(=O)(=O)NH(C1-C3 alkyl), -NR d’ S(=O)(=O)(C1-C6 alkyl), -P(=O)R e’ R f’ , sulfonic acid, -N=NPh, -Si(C1-C3 alkyl)3; wherein Ra’ , R b’ , R c’ , R d’ , R e’ , R f’ Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C(=O)(C1-C6 alkyl); wherein R 3’ , R 4’ Each is independently selected from hydrogen, C1-C6 alkyl, C(=O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy.

[0037] In certain embodiments, the substituents of ring B are selected from the following groups: hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo-C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)R b’ 、-OR c’ , C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group; wherein R a’ , R b’ , R c’ Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C(=O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy;

[0038] In certain embodiments, the substituents of ring B are selected from the following groups: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo-C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)R b’、-OR c’ .

[0039] In certain embodiments, R a’ , R b’ , R c’ , R d’ , R e’ , R f’ Each is independently selected from hydrogen, -OH, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -N(C1-C6 alkyl)2; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0040] In certain embodiments, R 3’ , R 4’ Selected from hydrogen, C1-C6 alkyl.

[0041] In certain embodiments, the 3-6 membered heterocyclyl is substituted by one or more groups selected from the group consisting of halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, and C1-C6 alkoxy.

[0042] In certain embodiments, the substituents of Ring B are selected from hydrogen, =0, C1-C6 alkyl.

[0043] In certain embodiments, Ring B is a pyridone or a pyridazinone.

[0044] In certain embodiments, Ring B is 1H-pyridin-2-one, having the structural formula:

[0045] In certain embodiments, Ring B is 2H-pyridazin-3-one, having the structural formula:

[0046] In certain embodiments, L is selected from -(CH2) m -W-(CH2) n -, wherein W is selected from -O-, -NH-, -N(Boc)-, -NHC(=O), -CH2-, -CH(C1-C6 alkyl)-, -C(C1-C6 alkyl)(C1-C6 alkyl)-, -C(=O)-, -C(=O)NH-, -N(C1-C6 alkyl)-, -S-, -S(=O)-, -S(=O)(=O)-, -P(=O)(OH)-, -OC(=O)-, -C(=O)O-, -NHS(=O)(=O)-, -S(=O)(=O)NH-, and m and n are each independently selected from 0, 1 or 2.

[0047] In certain embodiments, L is selected from the following structures: -O-CH2-, -NH-CH2-, -CH2-, -NH-, -NH-C(=O)-, -CH2-O-, -CH2-O-CH2-, -CH2-CH2-, -O-CH2-CH2-, -N(Boc)-CH2-.

[0048] In certain embodiments, L is selected from the following structures: -O-CH2-, -CH2-O-, -NH-CH2-, -CH2-NH-, -CH2-CH2-, -N(Boc)-CH2-.

[0049] In certain embodiments, L is selected from the following structures: -NH-CH2-, -CH2-CH2-, -N(Boc)-CH2-.

[0050] In certain embodiments, q is 0.

[0051] In certain embodiments, T is selected from -(CH-T 1 ) p -, wherein p is selected from an integer between 1 and 6 (e.g., 0, 1, 2, 3, 4, 5 or 6), T 1 Selected from: hydrogen, halogen, nitro, hydroxyl, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, mercapto, -S-(C1-C6 alkyl).

[0052] In certain embodiments, T 1 Selected from hydrogen, C1-C6 alkyl.

[0053] In certain embodiments, T is selected from -CH2-.

[0054] In certain embodiments, T is -CH(R 2 )-, R 2 Selected from hydrogen or C1-C6 alkyl.

[0055] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (I'), wherein R 1 , Q, L, Ring A and Ring B are as defined in any of the above items, respectively.

[0056]

[0057] R 2 Selected from hydrogen or C1-C6 alkyl.

[0058] In certain embodiments, R 2 For hydrogen.

[0059] In certain embodiments, R 2It is methyl.

[0060] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (II):

[0061]

[0062] in,

[0063] R 1 , Q, L are as defined in any of the above items;

[0064] R 2 Selected from hydrogen or C1-C6 alkyl.

[0065] In certain embodiments, R 2 For hydrogen.

[0066] In certain embodiments, R 2 It is methyl.

[0067] In certain embodiments, R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0068] In certain embodiments, R 5 , R 6 Each is independently selected from hydrogen, halogen, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4, -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0069] In certain embodiments, R 5 , R 6 Each is independently selected from hydrogen, halogen, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0070] In certain embodiments, R 7 Selected from hydrogen, C1-C6 alkyl (eg methyl).

[0071] In certain embodiments, R 7 For hydrogen.

[0072] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (II'):

[0073]

[0074] R 1 , R 2 , Q, L, R 5 , R 6 , R 7 As defined in any of the above items respectively.

[0075] In certain embodiments, R 5 , R 6 , R 7Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0076] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (II"):

[0077]

[0078] R 1 , R 2 , Q, L, R 5 , R 6 , R 7 As defined in any of the above items respectively.

[0079] In certain embodiments, R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0080] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (III):

[0081]

[0082] in,

[0083] R 1 , Q, L, R 5 , R 6 , R 7 as defined in any of the above items respectively;

[0084] R 2 Selected from hydrogen or C1-C6 alkyl.

[0085] In certain embodiments, R 2 For hydrogen.

[0086] In certain embodiments, R 2 It is methyl.

[0087] In certain embodiments, R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0088] In certain embodiments, R 5 , R 6 Each is independently selected from hydrogen, halogen, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4, -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0089] In certain embodiments, R 5 , R 6 Each is independently selected from hydrogen, halogen, halogenated C1-C6 alkyl, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0090] In certain embodiments, R 7 Selected from hydrogen, C1-C6 alkyl (eg methyl).

[0091] In certain embodiments, R 7 For hydrogen.

[0092] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (III'):

[0093]

[0094] R 1 , R 2 , Q, L, R 5 , R 6 , R 7 As defined in any of the above items respectively.

[0095] In certain embodiments, R 5 , R 6 , R 7Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0096] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (III"):

[0097]

[0098] R 1 , R 2 , Q, L, R 5 , R 6 , R 7 As defined in any of the above items respectively.

[0099] In certain embodiments, R 5 For hydrogen.

[0100] In certain embodiments, R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0101] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (V):

[0102]

[0103] Among them, R 1 , Q, L, R 5 , R 6 , R 7 As defined in any of the above;

[0104] X is C or N;

[0105] R 2 Selected from hydrogen or C1-C6 alkyl.

[0106] In certain embodiments, R 1 For hydrogen.

[0107] In certain embodiments, Q is -O-.

[0108] In certain embodiments, R 2 It is hydrogen or methyl.

[0109] In certain embodiments, R 2 For hydrogen.

[0110] In certain embodiments, R 2 It is methyl.

[0111] In certain embodiments, R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0112] In certain embodiments, R 5 , R 6Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered cycloalkyl, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-N(C1-C3 alkyl)(C1-C3 alkyl), cyano, -P(=O)(C1-C3 alkyl)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, -C(=O)(C1-C6 alkyl), -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), halogenated C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0113] In certain embodiments, the -CH2-(3-6 membered nitrogen-containing heterocyclyl) is

[0114] In certain embodiments, R 7 Selected from hydrogen, C1-C6 alkyl (eg methyl).

[0115] In certain embodiments, R 7 For hydrogen.

[0116] In certain embodiments, L is -CH2-CH2- or -NH-CH2-.

[0117] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (V'):

[0118]

[0119]

[0120] R 1 , R 2 , Q, L, X, R 5 , R 6 , R 7 As defined in any of the above items respectively.

[0121] In certain embodiments, R 5 For hydrogen.

[0122] In certain embodiments, R 6Selected from: hydrogen, halogen, 3-6 membered cycloalkyl, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-N(C1-C3 alkyl)(C1-C3 alkyl), cyano, -P(=O)(C1-C3 alkyl)(C1-C3 alkyl), -C(O-C1-C6 alkyl)=CH2, -C(=O)(C1-C6 alkyl), -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), halogenated C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0123] In certain embodiments, the -CH2-(3-6 membered nitrogen-containing heterocyclyl) is

[0124] In certain embodiments, R 7 Selected from hydrogen, C1-C6 alkyl (eg methyl).

[0125] In certain embodiments, the compounds of the invention have a structure as shown in Formula (V"):

[0126]

[0127] R 1 , R 2 , Q, L, X, R 5 , R 6 , R 7 As defined in any of the above items respectively.

[0128] In certain embodiments, R 5 For hydrogen.

[0129] In certain embodiments, R 6 Selected from: hydrogen, halogen, 3-6 membered cycloalkyl, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-N(C1-C3 alkyl)(C1-C3 alkyl), cyano, -P(=O)(C1-C3 alkyl)(C1-C3 alkyl), -C(O-C1-C6 alkyl)=CH2, -C(=O)(C1-C6 alkyl), -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), halogenated C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0130] In certain embodiments, the -CH2-(3-6 membered nitrogen-containing heterocyclyl) is

[0131] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (V'"):

[0132]

[0133] R 1 , R 2 , Q, L, X, R 5 , R 6 , R 7 As defined in any of the above items respectively.

[0134] In certain embodiments, R 5 For hydrogen.

[0135] In certain embodiments, R 6 Selected from: hydrogen, halogen, 3-6 membered cycloalkyl, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-N(C1-C3 alkyl)(C1-C3 alkyl), cyano, -P(=O)(C1-C3 alkyl)(C1-C3 alkyl), -C(O-C1-C6 alkyl)=CH2, -C(=O)-(C1-C6 alkyl), -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), halogenated C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

[0136] In certain embodiments, the -CH2-(3-6 membered nitrogen-containing heterocyclyl) is

[0137] In certain embodiments, R 6 Selected from: -Br, cyclopropyl, -CH2-N(CH3)(CH3), cyano, -P(=O)(CH3)(CH3), -C(OEt)=CH2, -C(=O)CH3, -SCH3, -S(=O)CH3, -S(=O)(=O)CH3, -CF3.

[0138] In certain embodiments, R 7 Selected from hydrogen or C1-C6 alkyl.

[0139] In certain embodiments, R 7 Selected from hydrogen.

[0140] In certain embodiments, R 7 Selected from methyl.

[0141] The hydroxyl groups and -NH- and -NH2 in the compounds of the present invention may be optionally protected by substituents, which may be hydroxyl or amino protecting groups commonly used in the art, such as Boc (tert-butyloxycarbonyl), TBDPS (tert-butyldiphenyl), etc. The compounds thus formed are also within the protection scope of the present invention.

[0142] The compounds of the present invention may have a structure selected from the following:

[0143]

[0144]

[0146] Pharmaceutical composition, use, and disease prevention / treatment methods

[0147] In one aspect, the present application provides a pharmaceutical composition comprising a compound of the present invention, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or a metabolite thereof; optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

[0148] The pharmaceutical composition of the present invention can be prepared into any pharmaceutically acceptable dosage form and applied to patients who need such treatment by oral, parenteral, rectal, pulmonary, topical or transdermal administration. When used for oral administration, it can be prepared into conventional solid preparations, such as tablets, capsules, pills, granules, etc.; it can also be prepared into oral liquid preparations, such as oral solutions, oral suspensions, syrups, etc. When made into oral preparations, suitable fillers, binders, disintegrants, lubricants, etc. can be added. When used for parenteral administration, it can be prepared into injections, including injections, sterile powders for injection and concentrated solutions for injection. When made into injections, conventional methods in the existing pharmaceutical field can be used for production. When preparing injections, additives may not be added, or suitable additives may be added according to the properties of the drug. When used for rectal administration, it can be prepared into suppositories, etc. When used for pulmonary administration, it can be prepared into inhalants or sprays, etc. When used for topical administration, it can be used in specific dosage forms and devices or prepared into creams, gels, etc. When used for transdermal administration, it can be prepared into patches.

[0149] In one aspect, the present application provides the use of the compounds of the present invention, their pharmaceutically acceptable salts, prodrugs, stereoisomers, tautomers, tautomers, hydrates, solvates, crystal forms, isotope labels or their metabolite forms for preparing drugs, which are used as TLR7 agonists for preventing or treating diseases associated with TLR7.

[0150] In one aspect, the present application provides a method for preventing or treating a TLR7-related disease in a subject, the method comprising administering to a subject in need thereof an effective amount of a compound of the present invention, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystalline form, isotope-labeled substance or a metabolite thereof.

[0151] In some embodiments, the disease associated with TLR7 is selected from:

[0152] (1) Tumors, including but not limited to: breast cancer, melanoma, meningioma, soft tissue sarcoma, salivary gland tumors, intraspinal tumors, mediastinal tumors, brain cancer, bone cancer, penile cancer, osteosarcoma, intracranial tumors, tongue cancer, maxillary sinus cancer, thyroid cancer, malignant lymphoma, multiple myeloma, pituitary adenoma, testicular tumors, non-Hodgkin's lymphoma, bladder cancer, leukemia, gastric cancer, nasopharyngeal cancer, laryngeal cancer, oral cancer, esophageal cancer, lung cancer, kidney cancer, cervical cancer, choriocarcinoma, vulvar cancer, skin cancer, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, rectal cancer, colorectal cancer, Kaposi's sarcoma, non-melanoma skin cancer (including squamous cell carcinoma and basal cell carcinoma), hemangioma, glioma, non-Hodgkin's lymphoma (diffuse large B-cell lymphoma, B-cell non-Hodgkin's lymphoma), colon cancer, liver cancer, neuroblastoma, glioblastoma;

[0153] (2) diseases associated with viral infection, including but not limited to dengue virus, yellow fever virus, West Nile virus, Japanese encephalitis virus, tick-borne encephalitis virus, Kunjin virus, Murray Valley encephalitis virus, St. Louis encephalitis virus, Omsk hemorrhagic fever virus, bovine viral diarrhea virus, Zika virus, HIV, HBV, HCV, HPV, RSV, SARS or influenza virus;

[0154] (3) Inflammatory diseases, respiratory diseases, skin diseases, gastrointestinal diseases, hepatobiliary diseases, nervous system diseases, musculoskeletal and connective tissue diseases, or autoimmune diseases, such as asthma, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome (ARDS), ulcerative colitis, Crohn's disease, bronchitis, dermatitis, actinic keratosis, allergic rhinitis, psoriasis, scleroderma, urticaria, rheumatoid arthritis, multiple sclerosis, lupus, drug-induced allergic reactions, and rheumatism.

[0155] In the method, the compound of the present invention acts as a TLR7 agonist, and further, can act as an immunomodulator.

[0156] The compounds of the present invention have an immunoenhancing effect. Therefore, in one aspect, the present application provides the use of the compounds of the present invention, their pharmaceutically acceptable salts, prodrugs, stereoisomers, tautomers, hydrates, solvates, crystal forms, isotope labels or their metabolites as vaccine adjuvants. In certain embodiments, the vaccine is used to prevent or treat diseases associated with any of the viral infections described above.

[0157] In one aspect, the present application provides use of the compound of the present invention, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope label or metabolite form thereof as a photodynamic therapy agent.

[0158] In one aspect, the present application provides the use of a compound of the present invention, a pharmaceutically acceptable salt, a prodrug, a stereoisomer, a tautomer, a hydrate, a solvate, a crystalline form, an isotope label or a metabolite thereof as a conjugated drug (e.g., an antibody immunostimulatory conjugate drug ISAC). An antibody immunostimulatory conjugate drug (ISAC) is a monoclonal antibody that combines an antibody with an immunostimulatory agent to enhance the immune response by triggering the production of proinflammatory cytokines and chemokines. For example, BDC-1001 is an ISAC that contains an antibody targeting HER2 and a TLR7 / 8 agonist, and is currently in clinical phase II for the treatment of metastatic breast cancer.

[0159] In the present application, the subject is preferably a mammal, such as bovine, equine, porcine, canine, feline, rodent, primate, such as human.

[0160] Preparation method

[0161] Route 1:

[0162]

[0163] Intermediate 1-1 is alkylated under alkaline conditions via intermediate 1-2 to generate 1-3, and intermediate 1-3 completes cyclization under alkaline conditions via nucleophilic attack to obtain the target product 1-4.

[0164] Among them, -X 1 Select from -OTs, -X 2 Selected from -OTBS, -COOMe, -CHO, and the remaining substituents are as described above.

[0165] Route 2:

[0166]

[0167] Intermediate 2-1 is alkylated under alkaline conditions via intermediate 2-2 to generate 2-3, and intermediate 2-3 completes cyclization under alkaline conditions via nucleophilic attack to obtain 2-4, and 2-4 undergoes a series of reactions to generate the target product 2-5.

[0168] Among them, -X 1 Select from -OTs, -X 2 Selected from -OTBS, -COOMe, -CHO, and the remaining substituents are as described above.

[0169] Definition of terms

[0170] In the present invention, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. In addition, the laboratory operation steps involved in this article are all conventional steps widely used in the corresponding fields. At the same time, in order to better understand the present invention, the definitions and explanations of relevant terms are provided below.

[0171] As used herein, the term "C1-C6 alkyl" refers to a group obtained by removing a hydrogen atom from a straight or branched alkane containing 1 to 6 carbon atoms, specific examples of which include but are not limited to: methyl, ethyl, propyl, n-butyl, isobutyl, isopropyl, tert-butyl, n-pentyl, n-hexyl. In the present invention, the preferred C1-C6 alkyl is a C1-C4 alkyl.

[0172] As used herein, the term "C2-C6 alkenyl" refers to a straight or branched hydrocarbon group containing at least one carbon-carbon double bond and having 2-6 carbon atoms. Specific examples include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1,3-butadienyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, and 1,4-hexadienyl. In the present invention, the preferred C2-C6 alkenyl is C2-C4 alkenyl.

[0173] As used herein, the term "C2-C6 alkynyl" refers to a straight or branched hydrocarbon group containing at least one triple bond and having 2-6 carbon atoms, and specific examples thereof include but are not limited to ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 1,3-butadiynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,4-hexadiynyl, and in the present invention, the preferred C2-C6 alkynyl is C2-C4 alkynyl. As used herein, the term "halogen" includes fluorine, chlorine, bromine and iodine. In the present invention, the preferred halogen is fluorine, chlorine, bromine, and more preferably bromine.

[0174] As used herein, the term "halogenated" means that hydrogen on a group or compound is replaced by one or more halogen atoms, including full halogenation and partial halogenation.

[0175] As used herein, the term "alkoxy" refers to a group formed in the form of alkyl-O-.

[0176] As used herein, the term "5-6 membered heteroaryl" refers to a monocyclic group with aromaticity containing 5-6 ring atoms and at least one ring atom being a heteroatom, wherein the heteroatom is a nitrogen atom, an oxygen atom and / or a sulfur atom. Specific examples include, but are not limited to, furyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl.

[0177] As used herein, the term "bicyclic heteroaryl" refers to a bicyclic group in which two rings (at least one of which has aromaticity) are fused together and at least one ring atom is a heteroatom, which is a nitrogen atom, an oxygen atom and / or a sulfur atom, and the nitrogen and / or sulfur are optionally substituted with oxo. Specific examples include, but are not limited to, benzopyrrole, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzimidazole, benzotriazole, indazole, purine, indole, quinoline, pteridine, isoquinoline, quinazoline, quinoxaline, 1,8-naphthyridine, 1,5-naphthyridine, 9H-purine, 7H-purine, indane, indoline, 2,3-dihydrobenzofuran, 1,2,3,4-tetrahydronaphthalene, 1,2,3,4-tetrahydroquinoline, 3,4-dihydro-1H-benzopyran, 1,4-benzodioxane, benzomorpholine, and 1,2,3,4-tetrahydroquinoxaline.

[0178] As used herein, the term "cycloalkyl" refers to a saturated or partially saturated monocyclic cyclic alkyl group that is not aromatic, preferably having 3 to 6 ring atoms, ie, a 3-6-membered saturated cycloalkyl group and a 3-6-membered partially saturated cycloalkyl group. Specific examples of 3-6 membered saturated cycloalkyl groups include, but are not limited to, cyclopropanyl (cyclopropyl), cyclobutanyl (cyclobutyl), cyclopentanyl (cyclopentyl), and cyclohexanyl (cyclohexyl); specific examples of 3-6 membered partially saturated cycloalkyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexa-1,3-dienyl, cyclohex-1,4-dienyl, cycloheptenyl, cyclohept-1,3-dienyl, cyclohept-1,4-dienyl, cyclohept-1,3,5-trienyl, cyclooctenyl, cyclooct-1,3-dienyl, cyclooct-1,4-dienyl, cyclooct-1,5-dienyl, cyclooct-1,3,5-trienyl, and cyclooctatetraenyl.

[0179] As used herein, the term "heterocyclyl" refers to a group obtained by removing a hydrogen atom from a saturated or partially saturated monocyclic or condensed ring compound containing 3-14 ring atoms and at least one heteroatom (e.g., 1, 2, 3, 4 or 5 heteroatoms). The term "3-6 membered nitrogen-containing heterocyclyl" refers to a heterocyclyl containing 3, 4, 5 or 6 ring atoms, 1, 2 or 3 of which are nitrogen atoms, and optionally, the heterocyclyl further contains 1 or 2 oxygen atoms or sulfur atoms.

[0180] As used herein, the term "pharmaceutically acceptable salt" refers to (i) a salt formed by an acidic functional group (e.g., -COOH) present in the compounds provided by the present invention and a suitable inorganic or organic cation (base), and includes, but is not limited to, alkali metal salts, such as sodium salts, potassium salts, lithium salts, etc.; alkaline earth metal salts, such as calcium salts, magnesium salts, etc.; other metal salts, such as aluminum salts, iron salts, zinc salts, copper salts, nickel salts, cobalt salts, etc.; inorganic base salts, such as ammonium salts; organic base salts, such as tert-octylamine salts, dibenzylamine salts, morpholine salts, glucosamine salts, phenylglycine alkyl ester salts, ethylenediamine salts, N-methylglucosamine salts, guanidine salts, diethylamine salts, triethylamine salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, chloroprocaine salts, procaine salts, diethanolamine salts, N-benzyl-phenethylamine salts, piperazine salts, tetramethylamine salts, tris(hydroxymethyl)aminomethane salts. And, (ii) salts formed by basic functional groups (e.g. -NH2) present in the compounds provided by the present invention and appropriate inorganic or organic anions (acids), and including but not limited to, hydrohalides, such as hydrofluorides, hydrochlorides, hydrobromides, hydroiodides, etc.; inorganic acid salts, such as nitrates, perchlorates, sulfates, phosphates, etc.; lower alkane sulfonates, such as methanesulfonates, trifluoromethanesulfonates, ethanesulfonates, etc.; aryl sulfonates, such as benzenesulfonates, p-toluenesulfonates, etc.; organic acid salts, such as acetates, malates, fumarates, succinates, citrates, tartrates, oxalates, maleates, etc.; amino acid salts, such as glycine, trimethylglycine, arginine, ornithine, glutamate, aspartate, etc.

[0181] As used herein, the term "pharmaceutically acceptable ester" refers to an ester formed by -COOH present in the compound provided by the present invention and a suitable alcohol, or an ester formed by -OH present in the compound provided by the present invention and a suitable acid (e.g., a carboxylic acid or an oxygen-containing inorganic acid). Suitable ester groups include, but are not limited to, formates, acetates, propionates, butyrates, acrylates, ethylsuccinates, stearic acid esters or palmitates. Esters can undergo hydrolysis reactions to generate corresponding acids or alcohols in the presence of acids or bases.

[0182] As used herein, the term "pharmaceutically acceptable amide" refers to an amide formed by -COOH present in the compounds provided herein and a suitable amine, or an amide formed by -NH- or -NH2 present in the compounds provided herein and a suitable acid (e.g., carboxylic acid, such as acetic acid).

[0183] As used herein, the term "solvate" refers to a substance formed by the association of the compounds of the present invention with solvent molecules. The solvent may be an organic solvent (e.g., methanol, ethanol, propanol, acetonitrile, etc.), for example, the compounds of the present invention may form ethanolates with ethanol. The compounds of the present invention may also form hydrates with water.

[0184] As used herein, the term "crystal form" refers to the crystal structure of a substance. When a substance crystallizes, various factors may affect the intramolecular or intermolecular bonding mode, causing molecules or atoms to be arranged differently in the lattice space, forming different crystal structures. The compounds of the present invention may exist in one crystal structure or in multiple crystal structures, i.e., have "polymorphism". The compounds of the present invention may exist in different crystal forms.

[0185] As used herein, the term "stereoisomer" includes conformational isomers and configurational isomers, wherein the configurational isomers mainly include cis-trans isomers and optical isomers. The compounds of the present invention can exist in the form of stereoisomers, and therefore encompass all possible stereoisomer forms, and any combination or any mixture thereof. For example, a single enantiomer, a single diastereomer or a mixture of the above. When the compounds of the present invention contain an olefin double bond, unless otherwise specified, it includes cis-isomers and trans-isomers, and any combination thereof.

[0186] As used herein, the term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom in a molecule between two positions, such as tautomers of the enol form and the keto form. If tautomers exist for the compounds of the present invention, they may exist in the form of a single tautomer or a mixture thereof, preferably in the form of a more stable tautomer as the main component.

[0187] As used herein, the term "prodrug" refers to a compound that can be converted into the present invention in the subject by reactions such as oxidation, reduction, hydrolysis, etc. The prodrug itself may or may not have the biological activity of the compound of formula (I) (e.g., regulating glucose and lipid metabolism activity, anti-inflammatory activity, antioxidant activity). For example, a compound of formula (I) including a hydroxyl group or a carboxyl group can be administered in the form of an ester, which is hydrolyzed and converted into a hydroxyl compound or a carboxyl compound in the body. Similarly, a compound of formula (I) including an amino group undergoes acylation, alkylation or phosphorylation to form, for example, eicosanoylamino, alanylamino, pivaloyloxymethylamino compounds for administration. Further information on the use of prodrugs can be found in Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T Higuchi and W Stella) and Bioreversible Carriers in Drug Design, Pergamon Press, 1987 (ed. EB Roche, American Pharmaceutical Association). Some examples of prodrugs according to the present invention include: (i) if the compound of formula (I) contains a carboxylic acid functional group (-COOH), then its esters include, for example, replacing hydrogen with a (C1-C8) alkyl group; (ii) if the compound of formula (I) contains an alcohol functional group (-OH), then its ethers include, for example, replacing hydrogen with a (C1-C6) alkanoyloxymethyl group; and (iii) if the compound of formula (I) contains a primary or secondary amino functional group (-NH2 or -NHR, wherein R is not H), then its amides include, for example, replacing hydrogen with a (C1-C6) alkanoyloxymethyl group. 10 )alkanoyl replaces one or two hydrogens. In addition, certain compounds of formula (I) may themselves act as prodrugs of other compounds of formula (I).

[0188] As used herein, the term "pharmaceutically acceptable carrier or excipient" refers to a carrier and / or excipient that is pharmacologically and / or physiologically compatible with a subject and an active ingredient, which is well known in the art (see, e.g., Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and includes, but is not limited to, disintegrants, binders, surfactants, glidants, lubricants, pH adjusters, ionic strength enhancers, agents that maintain osmotic pressure, agents that delay absorption, diluents, antioxidants, colorants, flavoring agents, preservatives, taste masking agents, etc. For example, non-limiting examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, cross-linked sodium carboxymethylcellulose, cross-linked polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropyl cellulose, starch, pregelatinized starch, and sodium alginate. Non-limiting examples of binders include microcrystalline cellulose, gelatin, sugar, polyethylene glycol, natural and synthetic gums, polyvinyl pyrrolidone, pregelatinized starch, hydroxypropyl cellulose and hydroxypropyl methylcellulose. Non-limiting examples of diluents include lactose (monohydrate, spray-dried monohydrate, anhydrous, etc.), mannitol, xylitol, glucose, sucrose, sorbitol, microcrystalline cellulose, starch and calcium hydrogen phosphate dihydrate. Non-limiting examples of surfactants include sodium lauryl sulfate and polysorbate 80. Non-limiting examples of glidants include silicon dioxide and talc. Non-limiting examples of lubricants include magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate and a mixture of magnesium stearate and sodium lauryl sulfate. Non-limiting examples of pH regulators include but are not limited to phosphate buffers. Ionic strength enhancers include but are not limited to sodium chloride. Reagents that maintain osmotic pressure include but are not limited to sugar, NaCl and the like. Reagents that delay absorption include but are not limited to monostearate and gelatin. Preservatives include, but are not limited to, various antibacterial and antifungal agents such as thimerosal, 2-phenoxyethanol, parabens, chlorobutanol, phenol, sorbic acid, and the like.

[0189] As used herein, the term "prevention" refers to a method implemented in order to prevent or delay the occurrence of a disease or illness or symptom in a subject. As used herein, the term "treatment" refers to a method implemented in order to obtain a beneficial or desired clinical result. For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, alleviating symptoms, reducing the scope of the disease, stabilizing (i.e., no longer worsening) the state of the disease, delaying or slowing the development of the disease, improving or alleviating the state of the disease, and alleviating symptoms (whether partially or completely), whether detectable or undetectable. In addition, "treatment" can also refer to, compared with the expected survival period (if not receiving treatment), extending the survival period.

[0190] As used herein, the term "effective amount" refers to an amount sufficient to obtain or at least partially obtain a desired effect. For example, a disease prevention (e.g., metabolic disease or the symptom associated with a metabolic disease) effective amount refers to an amount sufficient to prevent, prevent, or delay the occurrence of a disease (e.g., metabolic disease or the symptom associated with a metabolic disease); a disease treatment effective amount refers to an amount sufficient to cure or at least partially prevent the disease and its complications of a patient who has a disease. It is entirely within the capabilities of those skilled in the art to determine such an effective amount. For example, the effective amount for therapeutic use will depend on the severity of the disease to be treated, the overall state of the patient's own immune system, the patient's general condition such as age, weight and sex, the mode of administration of the drug, and other treatments administered simultaneously, etc. DETAILED DESCRIPTION

[0191] In the following examples, the structures of the compounds were determined by nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LCMS). NMR chemical shifts (δ) are expressed in 10 -6 The units are given in ppm. NMR measurements were performed using a Bruker AVANCE III 500 MHz NMR instrument, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD) and the internal standard being tetramethylsilane (TMS).

[0192] Analytical liquid chromatography: Analytical liquid chromatography is used to monitor the reaction system and analyze the purity of important intermediates and target products. Analytical liquid chromatography system: Agilent 1260 Infinity II (equipped with 1260 Binary Pump, Vial Sampler, MCT column oven, DAD WR detector); Chromatographic column: Waters XBridge BEH C18 3.5um, 4.6*150mm.

[0193] Preparative liquid chromatography uses preparative liquid phase to separate and purify a small amount of important intermediates and target products. Preparative liquid phase system: Agilent 1290 Infinity II (equipped with 1290 Prep Bin Pump, Prep ALS / FC injector / collector, 1260 VWD detector); C18 reverse phase silica gel column.

[0194] Liquid chromatography-mass spectrometry uses LCMS for conventional reaction system detection and molecular weight signal determination of target products. LC-MS system: Waters UPLCH-Class ultra-high performance liquid chromatography-QDa coupling instrument; chromatographic column: Waters XBridge BEH C181.7um, 2.1*75mm.

[0195] Liquid phase mobile phase ratio acidic conditions: A: 0.05% formic acid aqueous solution, B: acetonitrile; alkaline conditions: A: 0.05% ammonia aqueous solution, B: acetonitrile; neutral conditions: A: 5mM NH4HCO3 aqueous solution, B: acetonitrile. The above data are the default mobile phase ratios, which can be appropriately adjusted according to the properties of the actual samples when performing analysis and testing.

[0196] Rapid purification system Rapid purification system is used for rough preparation and purification of compounds in conventional reaction systems. The instrument model is Biotage Isolera One, and the normal phase column and reverse phase column used with it are purchased from Santai Tech. The common specifications of normal phase silica gel columns are 12g, 25g, 40g, 80g, 120g and 330g. The specific parameters are as follows: 40-63μm, The commonly used specifications of reverse phase C18 columns are 40g, 80g, 120g and 330g, and the specific parameters are as follows: 20-45μm,

[0197] Thin layer chromatography (TLC) silica gel plates are used to qualitatively analyze the components of a reaction system or to separate and purify a small amount of sample. The product used for qualitative analysis by TLC is Merck thin layer silica gel plates, with the following specifications: GF 254 , 20*20cm, glass plate, 0.25mm coating; The product used for separation and purification by thin layer chromatography is Leyan prepared silica gel plate, the specifications are as follows: GF 254 , 20*20cm, glass plate, 1mm coating.

[0198] Chemical reagents The known starting materials of the present invention can be synthesized by using or according to literature, patent methods, etc. known in the art, or can be purchased from chemical reagent companies such as Alfa Aesar, Acros, Sigma-Aldrich, TCI, Aicon, Anergy, Aladdin, J&K, Bid Pharmaceuticals, Leyan, McLean, Accela, Adamas, Xiens, and Inochem.

[0199] Abbreviations

[0200] Protecting Group Abbreviations:

[0201] Bn Benzyl

[0202] Boc tert-Butyloxycarbonyl

[0203] Cbz benzyloxycarbonyl

[0204] MOM Methoxymethyl

[0205] PMB p-Methoxybenzyl

[0206] TBDPS tert-Butyldiphenylsilyl

[0207] TBS tert-Butyldimethylsilyl

[0208] THP 2-Tetrahydropyranyl

[0209] Tos p-Toluenesulfonyl

[0210] Reagent Abbreviations:

[0211] ACN Acetonitrile

[0212] AIBN Azobisisobutyronitrile

[0213] Boc2O Di-tert-butyl dicarbonate

[0214] BPO Benzoyl Peroxide

[0215] tBuONa Sodium tert-butoxide

[0216] CAM Ammonium molybdate cerium sulfate color developer

[0217] CDI N,N'-Carbonyldiimidazole

[0218] m-CPBA m-Chloroperbenzoic acid

[0219] DABCO Triethylenediamine

[0220] DCE 1,2-Dichloroethane

[0221] DCM Dichloromethane

[0222] DEAD Diethyl azodicarboxylate

[0223] DIAD Diisopropyl azodicarboxylate

[0224] DIBAL-H Diisobutylaluminum hydride

[0225] DBU 1,8-diazobispiro[5.4.0]undec-7-ene DHP 3,4-Dihydro-2H-pyran

[0226] DIEA N,N-Diisopropylethylamine

[0227] DMAP 4-dimethylaminopyridine

[0228] DMF N,N-Dimethylformamide DMSO Dimethyl sulfoxide

[0229] EA Ethyl acetate

[0230] HMPA Hexamethylphosphoramide

[0231] HATU N,N,N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)

[0232] Urea hexafluorophosphate

[0233] LAH Lithium Aluminum Hydride

[0234] m-CPBA: meta-chloroperbenzoic acid

[0235] NBS N-Bromosuccinimide

[0236] PE Petroleum Ether

[0237] PMA Phosphomolybdic Acid

[0238] PyBOP 1H-Benzotriazol-1-yloxytripyrrolidino hexafluorophosphate TBAF Tetrabutylammonium fluoride

[0239] TBDPSCl tert-Butyldiphenylsilyl chloride

[0240] TBSCl tert-Butyldimethylsilyl chloride

[0241] TEA Triethylamine

[0242] TFA Trifluoroacetic acid

[0243] THF Tetrahydrofuran

[0244] TPSCl 2,4,6-Triisopropylbenzenesulfonyl chloride

[0245] TsCl p-Toluenesulfonyl chloride

[0246] TsOH p-Toluenesulfonic acid

[0247] Example 1: Synthesis of Compound C-1

[0248]

[0249] Step A: Under nitrogen protection at 0°C, a tetrahydrofuran solution of borane (20 mL, 1 M in THF, 20.00 mmol) was added dropwise to tetrahydrofuran (30 mL) in which 2-(3-(bromomethyl)phenyl)acetic acid (2.30 g, 10.04 mmol) was dissolved. After the addition was complete, the entire mixture was warmed to room temperature and stirred for 3 hours. Methanol was added to the reaction solution for quenching, and the reaction solution was concentrated under reduced pressure to obtain a yellow crude product. The crude product was separated by a silica gel column (PE:EA=85:15), and the reaction obtained a light yellow oily product 2-(3-(bromomethyl)phenyl)ethane-1-ol (2.30 g, 8.56 mmol, 85%). ESI-MS m / z=215.0[M+H] + ;Calcu.=214.0.

[0250] Step B: Potassium carbonate (2.39 g, 17.30 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2.05 g, 9.53 mmol) were added to N,N-dimethylformamide (20 mL) at room temperature in which 2-(3-(bromomethyl)phenyl)ethane-1-ol (1.47 g, 8.65 mmol) was dissolved. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a yellow solid. The residue was separated by silica gel column (PE:EA=85:15), and the reaction obtained the product 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)ethan-1-ol (888 mg, 2.93 mmol, 34%) as a white solid. ESI-MS m / z=304.0[M+H] + ;Calcu.=303.1.

[0251] Step C: At room temperature, triethylamine (1.7 mL, 11.99 mmol), 4-dimethylaminopyridine (32 mg, 0.264 mmol) and p-toluenesulfonyl chloride (1.14 g, 6.00 mmol) were added to dichloromethane (15 mL) in which 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)ethan-1-ol (888 mg, 2.93 mmol) was dissolved. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a yellow solid, and the residue was separated by silica gel column (PE:EA=85:15). The reaction obtained the product 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl-4-methylbenzenesulfonate (739 mg, 1.62 mmol, 55%) as a white solid. ESI-MS m / z=458.0[M+H] + ;Calcu.=457.1.

[0252] Step D: At room temperature, cesium carbonate (574 mg, 1.76 mmol) and methyl 6-oxo-1,6-dihydropyridine-3-carboxylate (224 mg, 1.46 mmol) were added to N,N-dimethylformamide (50 mL) in which 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl-4-methylbenzenesulfonate (739 mg, 1.62 mmol) was dissolved. After the addition, the whole mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid, and the residue was separated by silica gel column (DCM: MeOH = 90: 10). The reaction obtained the product 1-(3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl)-6-oxo-1,6-dihydropyridine-3-carboxylic acid methyl ester (500 mg, crude product) as a white solid. ESI-MS m / z = 439.0 [M+H] + ;Calcu.=438.1.

[0253] Step E: At room temperature, diisobutylaluminum hydride (3 mL, 1M hexane solution, 3.00 mmol) was added to toluene (25 mL) in which 1-(3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl)-6-oxo-1,6-dihydropyridine-3-carboxylic acid methyl ester (500 mg, crude product) was dissolved. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a yellow residue, which was separated by silica gel column (DCM:MeOH=90:10). The reaction gave a yellow solid product 1-(3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-ylmethyl)phenethyl)-5-hydroxymethylpyridin-2(1H)one (60 mg, 0.146 mmol, two-step yield 10%). ESI-MS m / z=411.0[M+H] + ;Calcu.=410.1.

[0254] Step F: At room temperature, cesium carbonate (143 mg, 0.438 mmol) and lithium iodide (59 mg, 0.438 mmol) were added to acetonitrile (5 mL) in which 1-(3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-ylmethyl)phenethyl)-5-hydroxymethylpyridin-2(1H)one (9 mg, 0.022 mmol) was dissolved. After the addition, the entire mixture was heated to 110°C and sealed under pressure and stirred for 36 hours. The reaction solution was concentrated under reduced pressure to obtain a white crude product, which was subjected to HPLC preparation, separation and purification. Preparation conditions: Waters XBridge RP18 (19mm×250mm, 5μm); A: 0.1% formic acid in water, B: MeCN; 18mL / min; 0~4min, 5% B; 4~16min, 5%~70% B; 16~17min, 70%~100% B; detector, UV254nm). The reaction yielded compound C-1 and white solid product (E)-1 4 -Amino-6 1 , 6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-phenylcyclooctan-6 2 -ketone (0.2 mg, 0.0005 mmol, 2%). 1H NMR(500MHz,DMSO-d6)δ7.50(s,1H),7.36-7.40(m,1H),7.31-7.16(m,3H),7.13-7.05(m,2H),6.53( s, 2H), 6.37 (d, J = 9.3Hz, 1H), 5.07 (s, 2H), 4.17 (s, 2H), 4.05 (t, J = 7.8Hz, 2H), 2.90 (t, J = 7.8Hz, 2H). ESI-MS m / z=375.0[M+H] + ;Calcu.=374.1.

[0255] Example 2: Synthesis of Compound C-2

[0256]

[0257] Step A: In a round-bottom flask, add methyl 6-oxo-1,6-dihydropyridazine-3-carboxylate (462 mg, 3 mmol), DMF (20 mL), cesium carbonate (977 mg, 3 mmol), and 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl-4-methylbenzenesulfonate (457 mg, 1 mmol) in sequence, and react at room temperature after all are added. After 18 hours, stirring was stopped, and water (20 mL) and a mixed solution of dichloromethane / methanol = 8 / 2 (16 mL dichloromethane + 4 mL methanol) were added in sequence. The organic layer was separated, and the aqueous layer was extracted with dichloromethane / methanol = 8 / 2 ((16 mL dichloromethane + 4 mL methanol) × 3). The organic layers were then combined, dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a solid crude product of 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid methyl ester (339 mg, crude product). The crude product was used directly in the next step without further separation. ESI-MS m / z = 440.0 [M+H] + ;Calcu.=439.1.

[0258] Step B: 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid methyl ester (150 mg, 0.34 mmol) and tetrahydrofuran (20 mL) were added to a sealed tube in sequence, and then placed in an ice-water bath at 0°C, and lithium aluminum hydride (52 mg, 1.36 mmol) was added. After all the addition, the mixture was reacted at 0°C. After 5 hours, 20 mL of saturated ammonium chloride solution and 20 mL of a mixed solution of dichloromethane / methanol = 16 / 4 (16 mL of dichloromethane + 4 mL of methanol) were added, and the organic layer was separated. The aqueous layer was extracted with dichloromethane / methanol = 16 / 4 ((16 mL of dichloromethane + 4 mL of methanol) × 3), and the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product (57 mg, crude product) of (1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid methyl ester) / (2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl)-6-(hydroxymethyl)pyridazin-3(2H)-one)=41 / 27 was directly used in the next step. ESI-MS m / z=412.0[M+H] + ;Calcu.=411.1.

[0259] Step C: Add the crude product (10 mg, 0.024 mmol), cesium carbonate (79 mg, 0.24 mmol), lithium iodide (40 mg, 0.24 mmol), and acetonitrile (3 mL) in the sealed tube in sequence. After all are added, tighten the sealed tube and place it in a 110°C oil bath for reaction. Remove from the oil bath after 8 hours and return to room temperature. Filter to remove cesium carbonate and potassium iodide, rinse with acetonitrile (10 mL), and dry the solvent under reduced pressure. The crude product is directly subjected to HPLC preparation and separation. Preparation conditions: YMC-Actus Triart C18 (20 mm × 250 mm, 5 μm); A: 0.05% ammonia water; B: MeCN; 18 mL / min; 0-5 min, 5% B; 5-20 min, 5%-20% B; detector, UV 254 nm. Obtain C-2, solid product (E)-1 4 -Amino-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -ketone (3 mg, 0.008 mmol, three-step yield 10%). 1H NMR(500MHz,DMSO-d6)δppm 7.92(s,1H)7.90-7.71(br,1H)7.66(s,1H)7.64-7.40(br,1H)7.25(d,J=10.0Hz,1H)7.12(d,J=10.0Hz,1H)7.06(t,J=7.5 Hz,2H)6.75(d,J=10.0Hz,1H)5.33(s,2H)5.27(s,2H)4.47(t,J=7.5Hz,2H)3.28(t,J=7.5Hz,2H); ESI-MSm / z=376.0[M+H] + ;Calcu.=375.1.

[0260] Example 3: Synthesis of Compound C-3 / C-5

[0261]

[0262] Step A: 6-oxo-1,6-dihydropyridazine-3-carboxylic acid methyl ester (706 mg, 4.6 mmol), cesium carbonate (1.49 g, 4.6 mmol), N,N-dimethylformamide (45 mL), 5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenethyl-4-methylbenzenesulfonate (820 mg, 1.5 mmol) were added to a round-bottom flask in sequence. After all the mixture was added, the mixture was reacted at room temperature. After 24 hours, stirring was stopped, and water (50 mL) and dichloromethane / methanol = 45 / 5 ((45+5) mL) were added in sequence. The organic layer was separated, and the aqueous layer was extracted with dichloromethane / methanol = 45 / 5 ((45+5) mL×3). The organic layers were then combined, dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a solid product, 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid methyl ester (578 mg, 1.1 mmol, 72%). ESI-MS m / z = 517.9 [M+H] + ;Calcu.=517.0.

[0263] Step B: 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid methyl ester (350 mg, 0.68 mmol), tetrahydrofuran (5 mL), and ethanol (45 mL) were added sequentially into a sealed tube. The sealed tube was then placed in an ice-water bath at 0°C, and sodium borohydride (128 mg, 3.4 mmol) and calcium chloride (749 mg, 6.8 mmol) were added. After all the ingredients were added, the reaction was continued at 0°C and naturally returned to room temperature. After 48 hours, water (50 mL) and a mixed solution of dichloromethane / methanol = 9 / 1 (45 mL dichloromethane + 5 mL methanol) were added, and the organic layer was separated. The aqueous layer was extracted with a mixed solution of dichloromethane / methanol = 9 / 1 ((45 mL dichloromethane + 5 mL methanol) × 3), and the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product of 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenethyl)-6-(hydroxymethyl)pyridazin-3(2H)-one (186 mg, crude product), which was used directly in the next step without further separation. ESI-MSm / z=489.8[M+H] + ;Calcu.=489.0.

[0264] Step C: Add 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenethyl)-6-(hydroxymethyl)pyridazine-3(2H)-one (180 mg, 0.37 mmol), cesium carbonate (1.20 g, 3.7 mmol), potassium iodide (609 mg, 3.7 mmol), and acetonitrile (25 mL) in a sealed tube. After all are added, tighten the sealed tube and place it in a 110°C oil bath for reaction. After 8 hours, remove from the oil bath and return to room temperature. Filter to remove cesium carbonate and potassium iodide, rinse with acetonitrile (20 mL), spin dry the solvent under reduced pressure, and send to preparative HPLC for separation. Preparation conditions: YMC-Actus Triart C18 (20mm×250mm, 5μm); A: 0.05% ammonia water; B: MeCN; 18mL / min; 0~3min, 5% B; 3~4min, 5%~10% B; 4~24min, 10%~50% B; detector, UV 254nm. Solid products C-5, (E)-1 were obtained. 4 -Amino-3 4 -Br-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6-ketone (90 mg, two-step yield 30%). ESI-MS m / z = 453.9 [M+H] + ;Calcu.=453.1.

[0265] Step D: Replace the nitrogen in the dry Schlenk tube three times, and then add (E)-1 4 -Amino-3 4 -Br-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -ketone (45mg, 0.1mmol), 1-((trifluoro-4-boryl)methyl)pyrrolidine potassium salt (57mg, 0.3mmol), tridibenzylideneacetone dipalladium (9mg, 0.01mmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (19mg, 0.04mmol), sodium carbonate (32mg, 0.3mmol), all added, nitrogen was replaced three times, 1,4-dioxane (1.6mL), water (0.4mL) were added in sequence. Then the Schlenk tube was placed in an 80℃ oil bath for reaction. After 2 hours, the oil bath was taken out, the mixture was returned to room temperature, the solvent was dried, and the mixture was sent to preparative HPLC for separation. Preparation conditions: Waters XBridge Prep Shield RP18 (19mm×250mm, 5μm); A: 0.1% formic acid in water; B: MeCN; 18mL / min; 0~3min, 5% B; 3~15min, 5%~30% B; detector, UV 254nm. Solid products C-3, (E)-1 were obtained. 4 -Amino-3 4 -(Pyrrolidine-1-methyl)-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -ketone (9 mg, 0.02 mmol, 20%). 1H NMR(500MHz,DMSO-d6)δppm 7.90(s,1H)7.89-7.75(br,1H)7.71(s,1H)7.68-7.50(br,1H)7.29(d,J=10.0Hz,1H)7.25-7.05(m,2H)6.79(d,J =5.0Hz,1H)5.35(s,2H)5.24(s,2H)4.53(t,J=5.0Hz,2H)3.50(s,2H)2.33-2.25(m,6H)1.70-1.60(m,4H); ESI-MS m / z=459.1[M+H] + ;Calcu.=458.2.

[0266] Example 4: Synthesis of Compound C-4

[0267]

[0268] Step A: Replace the nitrogen in the dry Schlenk tube three times, and then add (E)-1 4 -Amino-3 4 -Br-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -ketone (45 mg, 0.1 mmol), N,N-dimethyl-1-(trifluoro-λ 4 -boryl) methylamine potassium salt (49mg, 0.3mmol), tridibenzylideneacetone dipalladium (2mg, 0.002mmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (5mg, 0.01mmol), sodium carbonate (32mg, 0.3mmol), all added and replaced with nitrogen three times, 1,4-dioxane (1.6mL), water (0.4mL) were added in sequence. Then the Schlenk tube was placed in an 80℃ oil bath for reaction. After 3h, the oil bath was removed, the mixture was returned to room temperature, the solvent was dried by spin drying, and the mixture was sent for preparative HPLC separation. Preparation conditions: Waters XBridge Prep Shield RP18 (19mm×250mm, 5μm); A: 0.1% formic acid in water; B: MeCN; 18mL / min; 0~3min, 5% B; 3~23min, 5%~30% B; detector, UV 254nm. The solid product C-4, (E)-1 4 -Amino-3 4 -((dimethylamino)methyl)-6- 1,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -ketone (8 mg, 0.019 mmol, 19%). 1 H NMR(500MHz,DMSO-d6)δppm 7.90(s,1H),7.88-7.73(br,1H),7.70(s,1H),7.69-7.49(br,1H),7.29(d,J=10.0Hz,1H),7.27-7.03(m,2H),6. 78(d,J=10.0Hz,1H),5.34(s,2H),5.25(s,2H),4.53(t,J=5.0Hz,2H),3.50(t,J=10.0Hz,2H),2.25-2.08(m,8H). ESI-MS m / z=433.1[M+H] + ;Calcu.=432.2.

[0269] Example 5: Synthesis of Compound C-6 / C-10

[0270]

[0271] Step A: At room temperature, add concentrated sulfuric acid (98%) (0.5 mL, 9.335 mmol) to methanol (100 mL) in which (2-bromo-5-methylphenyl)acetic acid (15 g, 65.48 mmol) is dissolved. After the addition is complete, the entire mixture is heated to 80°C and stirred for 6 hours. The reaction solution is concentrated under reduced pressure to obtain a pale yellow solid-liquid mixture crude product, which is diluted with water and extracted three times with ethyl acetate (50 mL*3). The organic phases are combined, washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow oily liquid crude product 2-(2-bromo-5-methylphenyl)acetic acid methyl ester, which is directly used in the next step. ESI-MS m / z=244.8[M+H] + ;Calcu.=244.0.

[0272] Step B: At room temperature, dibenzoyl peroxide (1 g, 4.128 mmol) was added to carbon tetrachloride (100 mL) in which crude 2-(2-bromo-5-methylphenyl)acetate and N-bromosuccinimide (11.75 g, 66.02 mmol) were dissolved. After the addition, the whole mixture was heated to 80°C and stirred for 6 hours. The reaction solution was filtered through silica gel, washed with ethyl acetate, filtered and concentrated under reduced pressure to obtain a crude yellow oily liquid product. The residue was separated by silica gel column (SiO2, PE:EA=90:10), and the reaction obtained a colorless oily liquid product 2-(2-bromo-5-(bromomethyl)phenyl)acetate (16.28 g, 50.56 mmol, two-step yield 77%).

[0273] Step C: At 0°C and under N2 protection, diisobutylaluminum hydride (126mL, 126.0mmol) was added dropwise to toluene (200mL) in which methyl 2-(2-bromo-5-(bromomethyl)phenyl)acetate (16.28g, 50.56mmol) was dissolved. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was temperature controlled at 0°C, and then water was slowly added until no obvious bubbles appeared in the reaction solution, and then filtered through diatomaceous earth, extracted with ethyl acetate three times (50mL*3), the organic phases were combined, washed once with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a colorless oily liquid crude product. The residue was separated by silica gel column (SiO2, PE:EA=67:33), and the reaction obtained a white solid product 2-(2-bromo-5-(bromomethyl)phenyl)ethane-1-ol (9.04g, 30.75mmol, 61%). ESI-MS m / z=274.2[M-OH] + ;Calcu.=291.9.

[0274] Step D: Potassium carbonate (3.7 g, 26.77 mmol) was added to N,N-dimethylformamide (90 mL) and tetrahydrofuran (90 mL) in which 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (3 g, 17.69 mmol) and 2-(2-bromo-5-(bromomethyl)phenyl)ethane-1-ol (7.8 g, 26.53 mmol) were dissolved at room temperature. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a yellow oily crude product. The residue was separated by silica gel column (SiO2, DCM:MeOH=90:10). The reaction obtained a white solid product 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenyl)ethane-1-ol (1.94 g, 5.08 mmol, 29%). ESI-MS m / z=381.8[M+H] +;Calcu.=381.0.

[0275] Step E: At 0°C, p-toluenesulfonyl chloride (1.94 g, 10.18 mmol) was added in portions to a solution of 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenyl)ethane-1-ol (1.94 g, 5.08 mmol) and triethylamine (2.8 mL, 20.14 mmol) in dichloromethane (200 mL). After the addition was complete, the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a yellow oily crude product, and the residue was separated by silica gel column (SiO2, DCM:MeOH=10:1). The reaction gave a light yellow solid product 5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl-4-methylbenzenesulfonate (2.7 g, 5.03 mmol, 99%). ESI-MS m / z=535.8[M+H] + ;Calcu.=535.0.

[0276] Step F: At room temperature, cesium carbonate (911 mg, 2.796 mmol) was added to N,N-dimethylformamide (10 mL) in which 5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl-4-methylbenzenesulfonate (500 mg, 0.931 mmol) and 5-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-2(1H)-one (622 mg, 2.598 mmol) were dissolved. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was filtered through celite, washed with ethyl acetate, filtered, and concentrated under reduced pressure to obtain a brown oily liquid crude product. The residue was separated by silica gel column (SiO2, PE:EA=30:70 to EA:MeOH=90:10), and the reaction gave the product 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-5-((tert-butyldimethylsilyl)oxy)methyl ester)pyridin-2(1H)-one (130 mg, 0.215 mmol, 23%) as a white solid. ESI-MS m / z=603.0[M+H] + ;Calcu.=602.1.

[0277] Step G: At room temperature, tetrabutylammonium fluoride (0.6 mL, 0.60 mmol) was added to tetrahydrofuran (20 mL) in which 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-5-((tert-butyldimethylsilyl)oxy)methyl)pyridin-2(1H)-one (130 mg, 0.215 mmol) was dissolved. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was filtered through silica gel, washed with ethyl acetate:methanol=50:50, filtered and concentrated under reduced pressure to obtain a yellow solid crude product 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one, which was directly used in the next step. ESI-MS m / z=488.8[M+H] + ;Calcu.=488.0.

[0278] Step H: At room temperature, cesium carbonate (790 mg, 2.425 mmol) and lithium iodide (325 mg, 2.428 mmol) were added to acetonitrile (30 mL) in which the crude product of ((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one was dissolved. After the addition, the whole mixture was heated to 110°C and stirred for 16 hours. The reaction solution was filtered through diatomaceous earth, washed with ethyl acetate:methanol=50:50, filtered and concentrated under reduced pressure to obtain a yellow solid crude product. The residue was separated by TLC (SiO2, ethyl acetate:methanol=90:10with 1% triethylamine), and the reaction obtained a light yellow solid product (E)-1 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclononane-6 2 -ketone (70 mg, 0.154 mmol, two-step yield 72%). Take 10 mg of the product after scraping and perform HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 16 mL / min; 0-3 min, 5% B; 3-23 min, 5%-50% B; 23-30 min, 50%-100% B; detector, UV 254 nm. Compound C-10 and white solid product (E)-1 were obtained. 4 -Amino-34 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclononane-6 2 -ketone (1.61 mg, 0.003 mmol). 1 H NMR (500MHz, DMSO-d6) δppm8.33(d,J=2.2Hz,1H),7.96(s,1H),7.88(d,J=1.7Hz,1H),7.73(br s,1H),7.48(br s,1H),7.43(d,J=8.3Hz,1H),7.37(dd,J=9.5,2.2Hz,1H),6.95(dd,J=8.3,1.7Hz,1H),6.2 8(d,J=9.5Hz,1H),5.43(s,2H),5.07(s,2H),4.39(t,J=5.2Hz,2H),3.30(t,J=5.2Hz,2H). ESI-MS m / z=452.9[M+H] + ;Calcu.=452.1.

[0279] Step I: At room temperature and under N2 protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclononane-6 2-ketone (15mg, 0.033mmol), potassium trifluoro(pyrrolidin-1-ylmethyl)borate (38mg, 0.199mmol) in 1,4-dioxane (2mL), tridibenzylideneacetone dipalladium (6.1mg, 0.007mmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (12.6mg, 0.026mmol), sodium carbonate (21mg, 0.198mmol) and water (0.5mL) were added. After the addition was complete, the whole mixture was heated to 80°C and stirred for 4 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (19mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 16mL / min; 0-3min, 5% B; 3-18min, 5%-95% B; 18-19min, 95%-100% B; detector, UV 254nm. Compound C-6 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -(pyrrolidin-1-ylmethyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclosinphene-6 2 -ketone (4.10 mg, 0.009 mmol, 27%). 1 H NMR(500MHz,DMSO-d6)δppm 8.22(d,J=2.2Hz,1H),7.94(s,1H),7.72(br s,1H),7.62(s,1H),7.46(br s,1H),7.34(dd,J=9.3,2.2Hz,1H),7.09(d,J=8.0Hz,1H),6.92(dd,J=8.0Hz,1.1Hz,1H),6.25(d,J=9.3Hz,1H) ,5.40(s,2H),5.06(s,2H),4.33(t,J=5.4Hz,2H),3.40(s,2H),3.25(t,J=5.4Hz,2H),2.33(s,4H),1.64(s,4H). ESI-MS m / z=458.3[M+H] + ;Calcu.=457.2.

[0280] Example 6: Synthesis of Compound C-7

[0281]

[0282] Step A: At room temperature and under N2 protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclononane-6 2 -ketone (25mg, 0.055mmol), potassium (morpholin-4-yl)methyl trifluoroborate (69mg, 0.288mmol) in 1,4-dioxane (4mL), tridibenzylideneacetone dipalladium (5mg, 0.005mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (12mg, 0.025mmol), sodium carbonate (35mg, 0.330mmol) and water (1mL) were added. After the addition was complete, the whole mixture was heated to 80°C and stirred for 4 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (19mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 18mL / min; 0-3min, 10% B; 3-3.5min, 10%-20% B; 3.5-15min, 20%-50% B; 15-15.5min, 50%-100% B; detector, UV 254nm. Compound C-7 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -(Morpholinemethyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclosinphene-6 2 -ketone (2.99 mg, 0.006 mmol, 11%). 1H NMR (500MHz, DMSO-d6) δppm 8.19 (d, J=1.8Hz, 1H), 7.94 (s, 1H), 7.71 (br s, 1H), 7.62 (s, 1H), 7.46 (br s,1H),7.34(dd,J=9.4,2.4Hz,1H),7.07(d,J=7.6Hz,1H),6.92(d,J=7.6Hz,1H),6.25(d,J=9.4Hz, 1H), 5.40 (s, 2H), 5.06 (s, 2H), 4.34 (t, J = 5.2Hz, 2H), 3.50 (s, 4H), 3.25-3.30 (m, 4H), 2.25 (s, 4H). ESI-MSm / z=474.3[M+H] + ;Calcu.=473.2.

[0283] Example 7: Synthesis of Compound C-8

[0284]

[0285] Step A: At room temperature and under N2 protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclononane-6 2 -ketone (25mg, 0.055mmol), potassium dimethylaminomethyl trifluoroborate (55mg, 0.288mmol) in 1,4-dioxane (4mL), tridibenzylideneacetone dipalladium (5mg, 0.005mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (12mg, 0.025mmol), sodium carbonate (35mg, 0.330mmol) and water (1mL) were added. After the addition, the whole mixture was heated to 80°C and stirred for 4 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (19mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 18mL / min; 0-3min, 10% B; 3-3.5min, 10%-20% B; 3.5-16min, 20%-60% B; 16-16.5min, 60%-100% B; detector, UV 254nm. Compound C-8 and white solid product (E)-1 were obtained. 4-Amino-3 4 -((dimethylamino)methyl)-6- 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclosinphene-6 2 -ketone (3.43 mg, 0.008 mmol, 14%). 1 H NMR (500MHz, DMSO-d6) δppm 8.20 (d, J=2.3Hz, 1H), 7.94 (s, 1H), 7.71 (br s, 1H), 7.64 (s, 1H), 7.45 (br s,1H),7.34(dd,J=9.4,2.3Hz,1H),7.04(d,J=7.8Hz,1H),6.90(d,J=7.8Hz,1H),6.24(d,J=9.4Hz,1 H), 5.41 (s, 2H), 5.06 (s, 2H), 4.33 (t, J = 5.4Hz, 2H), 3.25 (t, J = 5.4Hz, 2H), 3.18 (s, 2H), 2.05 (s, 6H). ESI-MS m / z=432.3[M+H] + (C 23 H 26 N7O2); Calcu.=431.2.

[0286] Example 8: Synthesis of Compound C-9

[0287]

[0288] Step A: At room temperature and under N2 protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclononane-6 2-ketone (25mg, 0.055mmol), 1-methyl-4-trifluoroborylmethylpiperazine potassium (73mg, 0.288mmol) in 1,4-dioxane (4mL), tridibenzylideneacetone dipalladium (5mg, 0.005mmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (12mg, 0.025mmol), sodium carbonate (35mg, 0.330mmol) and water (1mL) were added. After the addition, the whole mixture was heated to 80°C and stirred for 4 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (19mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 18mL / min; 0-3min, 10% B; 3-3.5min, 10%-20% B; 3.5-16min, 20%-50% B; 16-16.5min, 50%-100% B; detector, UV 254nm. Compound C-9 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -((4-methylpiperazin-1-yl)methyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclosinphene-6 2 -ketone (3.75 mg, 0.008 mmol, 14%). 1 H NMR(500MHz,DMSO-d6)δppm 8.18(s,1H),7.94(s,1H),7.71(br s,1H),7.59(s,1H),7.46(br s,1H),7.35(d,J=9.2Hz,1H),7.05(d,J=7.8Hz,1H),6.93(d,J=7.8Hz,1H),6.26(d,J=9.2Hz,1H),5.40(s,2 H), 5.06 (s, 2H), 4.32 (t, J = 5.2Hz, 2H), 3.27 (s, 2H), 3.23 (s, J = 5.2Hz, 2H), 2.18-2.34 (m, 8H), 2.11 (s, 3H). ESI-MS m / z=487.4[M+H] + ;Calcu.=486.2.

[0289] Example 9: Synthesis of Compound C-11

[0290]

[0291] Step A: At room temperature, concentrated sulfuric acid (0.5 mL, 9.335 mmol) was added dropwise to methanol (50 mL) in which 2-(3-fluoro-5-methylphenyl)acetic acid (5 g, 29.73 mmol) was dissolved. After the addition, the whole mixture was stirred at 100°C for 3 hours. The reaction solution was adjusted to neutrality with sodium hydroxide solution and then concentrated. Water (50 mL) was added to the reaction solution, and it was extracted three times with ethyl acetate (50 mL*3). The organic phases were combined, washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a colorless oily crude product 2-(3-fluoro-5-methylphenyl)acetic acid methyl ester (5.14 g, 28.21 mmol, 95%).

[0292] Step B: At room temperature, dibenzoyl peroxide (1.05 g, 4.335 mmol) and N-bromosuccinimide (9.24 g, 51.90 mmol) were added to carbon tetrachloride (75 mL) in which 2-(3-fluoro-5-methylphenyl)acetate (7.88 g, 43.25 mmol) was dissolved. After the addition, the whole mixture was stirred at 90°C for 6 hours. The reaction solution was filtered and concentrated under reduced pressure to obtain a pale yellow oily crude product. The crude product was separated by silica gel column (SiO2, PE:EA=90:10), and the reaction obtained a colorless oily product 2-(3-(bromomethyl)-5-fluorophenyl)acetate (5.9 g, 22.60 mmol, 52%).

[0293] Step C: Under nitrogen protection at 0°C, diisobutylaluminum hydride (56.5 mL, 56.50 mmol) was added dropwise to toluene (100 mL) in which 2-(3-(bromomethyl)-5-fluorophenyl)acetic acid methyl ester (5.9 g, 22.60 mmol) was dissolved. After the addition, the whole mixture was stirred at room temperature for 2 hours. Methanol was slowly added to the reaction solution until no bubbles were generated, the solid was filtered out, and the organic phase was concentrated under reduced pressure to obtain a white crude product. The crude product was separated by silica gel column (SiO2, PE:EA=83:17), and the reaction obtained a white solid product 2-(3-(bromomethyl)-5-fluorophenyl)ethane-1-ol (2.5 g, 10.73 mmol, 47%).

[0294] Step D: At room temperature, 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.2 g, 7.151 mmol) and cesium carbonate (3.5 g, 10.73 mmol) were added to N,N-dimethylformamide (36 mL) in which 2-(3-(bromomethyl)-5-fluorophenyl)ethane-1-ol (2 g, 8.581 mmol) was dissolved, and the whole mixture was stirred overnight at room temperature. The reaction solution was filtered and concentrated under reduced pressure to obtain a yellow oily crude product. The crude product was separated by silica gel column (SiO2, DCM:MeOH=97:3) and the reaction obtained a yellow solid product 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenyl)ethane-1-ol (600 mg, 1.865 mmol, 26%). ESI-MS m / z=321.9[M+H] + ;Calcu.=321.1.

[0295] Step E: At 0°C, triethylamine (1 mL, 7.459 mmol) and 4-dimethylaminopyridine (22.8 mg, 0.187 mmol) were added to dichloromethane (10 mL) in which 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenyl)ethane-1-ol (600 mg, 1.865 mmol) was dissolved, followed by addition of p-toluenesulfonyl chloride (711 mg, 3.730 mmol) in batches. After addition, the whole mixture was stirred at room temperature overnight. The mixture was concentrated under reduced pressure to obtain a crude yellow oily product. The crude product was separated by silica gel column (SiO2, PE:EA=1:1), and the reaction gave a light yellow solid product 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenethyl-4-methylbenzenesulfonate (500 mg, 1.051 mmol, 56%). ESI-MS m / z=476.2[M+H] + ;Calcu.=475.1.

[0296] Step F: At room temperature, 5-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-2(1H)-one (229 mg, 0.96 mmol) and cesium carbonate (467 mg, 1.433 mmol) were added to N,N-dimethylformamide (10 mL) in which 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenethyl-4-methylbenzenesulfonate (500 mg, 1.051 mmol) was dissolved, and the whole mixture was stirred overnight at room temperature. The reaction solution was filtered and concentrated under reduced pressure to obtain a yellow oily crude product. The crude product was separated by silica gel column (SiO2, DCM:MeOH=90:10). The reaction gave a pale yellow solid product, 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenethyl)-5-((tert-butyldimethylsilyl)oxy)methyl (200 mg, 0.368 mmol, 39%). ESI-MS m / z=543.1[M+H] + ;Calcu.=542.2.

[0297] Step G: At room temperature, tetrabutylammonium fluoride (0.48 mL, 0.48 mmol) was added dropwise to tetrahydrofuran (10 mL) in which 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenylethyl)-5-((tert-butyldimethylsilyl)oxy)methyl (130 mg, 0.239 mmol) was dissolved, and the whole mixture was stirred at room temperature for 1.5 hours. The reaction solution was concentrated under reduced pressure to obtain a yellow crude product 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenylethyl)-5-(hydroxymethyl)pyridin-2(1H)-one, which was directly used for the next step without separation. ESI-MSm / z=429.3[M+H] + ;Calcu.=428.1.

[0298] Step H: At room temperature, cesium carbonate (228 mg, 0.700 mmol) and lithium iodide (94 mg, 0.70 mmol) were added to acetonitrile (3 mL) in which crude 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenylethyl)-5-(hydroxymethyl)pyridin-2(1H)-one was dissolved, and the whole mixture was stirred at 110°C for 16 hours. Subsequently, the mixture was concentrated under reduced pressure to obtain a yellow crude product, which was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (10 mm×150 mm, 5 μm); A: 5 mM NH3 in water, B: MeCN; 5 mL / min; 0-3 min, 5% B; 3-15 min, 5%-50% B; detector, UV 254 nm). The reaction gave compound C-11 and white solid product (E)-1 4 -Amino-3 5 -Fluoro-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzoic acid cyclosinphen-6 2 -ketone (5.10 mg, 0.013 mmol, two-step yield 19%). 1 H NMR(DMSO-d6,500MHz)δ8.07(s,1H),7.98(s,1H),7.76(br s,1H),7.55(s,2H),7.33(dd,J=9.3,2.1Hz,1H),6.81(br d,J=9.5Hz,1H),6.51(br d, J=9.5Hz, 1H), 6.24 (d, J=9.3Hz, 1H), 5.45 (s, 2H), 5.04 (s, 2H), 4.31 (br t, J=5.6Hz, 2H), 3.20 (br t, J=5.6Hz, 2H). ESI-MS m / z=393.2[M+H] + ;Calcu.=392.1.

[0299] Example 10: Synthesis of Compound C-12

[0300]

[0301] Step A: At room temperature, add azobisisobutyronitrile (1.81 g, 11.01 mmol) to carbon tetrachloride (150 mL) in which 1-methoxy-3,5-dimethylbenzene (15 g, 110.1 mmol) and N-bromosuccinimide (20.58 g, 115.6 mmol) are dissolved. After the addition, the whole mixture is heated to 80°C and stirred for 8 hours. The reaction solution is cooled to room temperature and concentrated under reduced pressure to obtain a crude product. The crude product is separated by a silica gel column (SiO2, 100% PE). The reaction obtains a colorless oily product 3-(bromomethyl)-1-methoxy-5-toluene (17.25 g, 73%). 1 H NMR (500MHz, DMSO-d6) δppm 6.83 (s, 1H), 6.81 (d, J = 1.8Hz, 1H), 6.71 (s, 1H), 4.62 (s, 2H), 3.73 (s, 3H), 2.26 (s, 3H).

[0302] Step B: At room temperature, trimethylsilyl cyanide (4.72 g, 47.61 mmol) was added to acetonitrile (50 mL) in which 3-(bromomethyl)-1-methoxy-5-toluene (5.12 g, 23.80 mmol) and potassium carbonate (4.93 g, 35.71 mmol) were dissolved. After the addition, the whole mixture was heated to 60°C and stirred for 12 hours. The reaction solution was cooled to room temperature, and then 1M sodium hydroxide was added to make it alkaline. It was extracted with ethyl acetate three times (50 mL x 3), the organic phases were combined, washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column (SiO2, PE:EA=90:10). The reaction obtained a colorless liquid product (5-methoxy-3-methylphenyl)acetonitrile (2.52 g, 66%). 1 H NMR (500MHz, DMSO-d6) δppm 6.70-6.75 (m, 3H), 3.95 (s, 2H), 3.74 (s, 3H), 2.28 (s, 3H).

[0303] Step C: At room temperature, 3M potassium hydroxide (9.036mL, 27.11mmol) was added to ethanol (18mL) in which (5-methoxy-3-methylphenyl)acetonitrile (1.90g, 11.79mmol) was dissolved. After the addition, the whole mixture was heated to 80°C and stirred for 12 hours. The reaction solution was concentrated under reduced pressure, extracted with EA three times (10mL x 3), the aqueous phase was adjusted to pH 6 with 1M hydrochloric acid, extracted with (DCM:MeOH=6:1) three times (20mL x 3), the organic phases were combined, washed once with saturated brine (10mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product (5-methoxy-3-methylphenyl)acetic acid (1.60g, crude product). It was directly used in the next step.

[0304] Step D: At room temperature, add azobisisobutyronitrile (0.13 g, 0.777 mmol) to carbon tetrachloride (14 mL) in which (5-methoxy-3-methylphenyl)acetic acid (1.40 g, 7.769 mmol) and N-bromosuccinimide (1.38 g, 7.769 mmol) are dissolved. After the addition, the entire mixture is heated to 100°C and stirred for 12 hours. The reaction solution is cooled to room temperature and concentrated under reduced pressure to obtain a crude product. The crude product is separated on a silica gel column (SiO2, DCM: MeOH = 97: 3) to obtain a colorless oily product (3-(bromomethyl)-5-methoxyphenyl)acetic acid (2.00 g, crude product). It is directly used in the next step reaction.

[0305] Step E: At 0°C, slowly drop borane tetrahydrofuran (15.4 mL, 15.4 mmol) into tetrahydrofuran (20 mL) in which (3-(bromomethyl)-5-methoxyphenyl)acetic acid (2.00 g, 7.72 mmol) was dissolved. After the addition, the entire mixture was slowly warmed to room temperature and stirred overnight. The reaction solution was placed in an ice bath and methanol was slowly added to quench. Concentrate under reduced pressure to obtain a crude product. The crude product was separated by a silica gel column (SiO2, PE:EA=75:25). The reaction gave a colorless oily product 2-(3-(bromomethyl)-5-methoxyphenyl)ethane-1-ol (550 mg, three-step yield 22%). 1 H NMR(500MHz,DMSO-d6)δppm 6.87(s,1H),6.84-6.85(m,1H),6.73-6.76(m,1H),4.63(s,2H),3.74(s,3H),3.59(t,J=7.0Hz,2H),2.68(t,J=7.0Hz,2H).

[0306] Step F: At room temperature, 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (399 mg, 1.628 mmol) was added to N,N-dimethylformamide (2.5 mL) and tetrahydrofuran (2.5 mL) in which 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (230 mg, 1.356 mmol) and potassium carbonate (375 mg, 2.713 mmol) were dissolved. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was dissolved in methanol (2 mL) and subjected to reverse phase separation. The chromatographic column was Santai Sepa Flash C18 (Spherical C18, 20-45 μm, 80g); mobile phase: water (containing 0.05% TFA) and acetonitrile; flow rate: 40 ml / min; gradient: acetonitrile increased from 0% to 100% in 30 minutes; detection wavelength: 254nm. Freeze-dried under reduced pressure to obtain a yellow solid product 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxyphenyl)ethane-1-ol (80mg, 16%). ESI-MS m / z=333.9[M+H] + ;Calcu.=333.1.

[0307] Step G: At 0°C, p-toluenesulfonyl chloride (91.4 mg, 0.479 mmol) was added to dichloromethane (2 mL) in which 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxyphenyl)ethane-1-ol (40 mg, 0.12 mmol), triethylamine (0.1 mL, 0.719 mmol) and 4-dimethylaminopyridine (1.46 mg, 0.012 mmol) were dissolved. After the addition was complete, the whole mixture was slowly heated to room temperature and stirred for 3 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated on a silica gel column (SiO2, PE:EA=75:25). The reaction gave a white solid product, 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxyphenethyl-4-methylbenzenesulfonate (30 mg, 46%). ESI-MS m / z = 487.9 [M+H] + ;Calcu.=487.1.

[0308] Step H: At room temperature, 6-oxo-1H-pyridine-3-carboxaldehyde (22.7 mg, 0.183 mmol) was added to N,N-dimethylformamide (2 mL) in which 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxyphenethyl-4-methylbenzenesulfonate (30 mg, 0.061 mmol) and cesium carbonate (60 mg, 0.183 mmol) were dissolved. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a white solid crude product 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxyphenethyl)-6-oxo-1,6-dihydropyridine-3-carboxaldehyde (40 mg, crude product). ESI-MS m / z=438.9[M+H] + ;Calcu.=438.1.

[0309] Step I: Sodium borohydride (10.3 mg, 0.27 mmol) was added to ethanol (2 mL) in which 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxyphenethyl)-6-oxo-1,6-dihydropyridine-3-carbaldehyde (40 mg, crude product) was dissolved at 0°C. After the addition, the whole mixture was slowly returned to room temperature and stirred overnight. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was dissolved in methanol (2 mL) and subjected to reverse phase separation. The chromatographic column was Santai SepaFlash C18 (Spherical C18, 20-45 μm, 40g); mobile phase: water (containing 0.05% TFA) and acetonitrile; flow rate: 40 ml / min; gradient: acetonitrile increased from 0% to 100% in 30 minutes; detection wavelength: 254nm. Freeze-dried under reduced pressure to obtain the product 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxyphenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one (20mg, two-step yield 72%) as a white solid. ESI-MS m / z=440.9[M+H] + ;Calcu.=440.1.

[0310] Step J: At room temperature, lithium iodide (65 mg, 0.49 mmol) was added to acetonitrile (2 mL) in which 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxyphenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one (20 mg, 0.049 mmol) and cesium carbonate (159 mg, 0.49 mmol) were dissolved. After the addition, the whole mixture was heated to 110°C and stirred for 12 hours. The reaction solution was cooled to room temperature, filtered, and then methanol (5 mL) was added to wash the filter cake. The filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was subjected to HPLC preparation and separation, preparation conditions: Waters XBridge Prep Shield RP18 (19mm×250mm, 5μm); A: 5mM NH4HCO3 in water, B: MeCN; 18mL / min; 0~3min, 10% B; 3~3.5min, 10%~20% B; 3.5~18min, 20%~80% B; detector, UV 254nm). After lyophilization under reduced pressure, compound C-12 and white solid product (E)-1 were obtained. 4 -Amino-3 5 -Methoxy-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (2.19 mg, 11%). 1 H NMR (500MHz, DMSO-d6) δppm8.06 (d, J=1.4Hz, 1H), 7.96 (s, 1H), 7.71 (br s, 1H), 7.46 (br s,1H),7.33(dd,J=9.1,2.1Hz,1H),7.25(s,1H),6.52(s,1H),6.36(s,1H),6.24(d,J=9. 3Hz, 1H), 5.38 (s, 2H), 5.04 (s, 2H), 4.28 (t, J = 5.5Hz, 2H), 3.55 (s, 3H), 313-3.16 (m, 2H). ESI-MS m / z=405.0[M+H] + ;Calcu.=404.2.

[0311] Example 11: Synthesis of Compound C-13

[0312]

[0313] Step A: At 20°C, sulfuric acid (0.65 mL, 12.18 mmol) was slowly added dropwise to methanol (10 mL) in which 2-(3,5-dimethylphenyl)acetic acid (20.0 g, 121.8 mmol) was dissolved, and the whole mixture was stirred at 70°C for 2 hours. The reaction solution was directly concentrated under reduced pressure to remove a large amount of methanol, and then water (50 mL) was added, and extracted twice with ethyl acetate (100 mL*2), the two organic phases were combined, washed once with saturated sodium chloride aqueous solution (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow oil, which was separated by silica gel column (SiO2, PE:EA=83:17). The reaction obtained a yellow oily product, 2-(3,5-dimethylphenyl)acetic acid methyl ester (21.0 g, 97%). 1 H NMR (500MHz, DMSO-d6) δppm 6.88 (s, 1H), 6.85 (s, 2H), 3.60 (s, 3H), 3.57 (s, 2H), 2.24 (s, 6H).

[0314] Step B: At 25°C, add azobisisobutyronitrile (954 mg, 5.813 mmol) to 1,2-dichloroethane (100 mL) in which methyl 2-(3,5-dimethylphenyl)acetate (10.36 g, 58.13 mmol) is dissolved, then the whole mixture is heated to 80°C, and N-bromosuccinimide (20.69 g, 116.26 mmol) is slowly added in batches at 80°C. After the addition, the whole mixture is heated to 90°C and stirred at 90°C for 0.5 hours. The reaction solution is cooled to room temperature, dichloromethane (100 mL) is added, and then washed once with saturated sodium bicarbonate aqueous solution (100 mL), washed once with saturated sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a solid residue. The solid residue is separated by silica gel column (SiO2, PE:EA=83:17). The reaction afforded a light yellow transparent oily product, methyl 2-(3,5-bis(bromomethyl)phenyl)acetate (10.15 g, 52%). 1 HNMR (500MHz, DMSO-d6) δppm7.43 (s, 1H), 7.29 (d, J = 1.4Hz, 2H), 4.69 (s, 4H), 3.70 (s, 2H), 3.62 (s, 3H).

[0315] Step C: At -30°C and nitrogen environment, diisobutylaluminum hydride (45.3mL, 45.3mmol) was slowly added dropwise to dichloromethane (100mL) in which methyl 2-(3,5-bis(bromomethyl)phenyl)acetate (10.15g, 30.21mmol) was dissolved. After the addition was complete, the entire mixture was stirred at 0-5°C and nitrogen environment for 2 hours. At 0-5°C, saturated aqueous ammonium chloride solution was slowly added to the reaction solution until no bubbles were generated. After the addition was complete, it was stirred for 30 minutes, then filtered through diatomaceous earth, the filtrate was collected, the filter cake was washed twice with dichloromethane (200mL*2), the filtrate was collected, washed once with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a solid residue. The solid residue was separated by silica gel column (SiO2, PE:EA=75:25). The reaction gave bis(bromomethyl)phenyl)ethane-1-ol (6.35 g, 68%) as a pale yellow solid product. 1 H NMR (500MHz, DMSO-d6) δppm 7.35 (s, 1H) 7.24 (d, J = 1.2 Hz, 2H), 4.67 (s, 4H), 3.60 (t, J = 6.9 Hz, 2H), 2.71 (t, J = 6.9 Hz, 2H).

[0316] Step D: At 40°C, 2-(3,5-bis(bromomethyl)phenyl)ethane-1-ol (6.35 g, 20.62 mmol) and potassium carbonate (8.55 g, 61.85 mmol) were dissolved in N,N-dimethylformamide (3 mL), and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2.80 g, 16.49 mmol) was slowly added in three batches, and the whole mixture was stirred at 40°C for 0.5 hours, and then water (3 mL) was added to the mixture, and then the whole mixture was stirred at 40°C for 16 hours. The reaction solution was directly concentrated under reduced pressure to obtain a solid residue. The solid residue was separated by silica gel column (SiO2, DCM:MeOH=94:6). The reaction afforded the product 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(hydroxymethyl)phenyl)ethan-1-ol as a yellow oil (500 mg, 7%). 1H NMR(500MHz,DMSO-d6)δppm 8.31(br s,1H),8.18(br s,1H),8.12(s,1H),7.05(s,1H),6.93-6.95(m,2H),5.39(s,2H),5.11(br s,1H),4.59(br s, 1H), 4.39 (s, 2H), 3.55 (t, J = 7.1Hz, 2H), 2.66 (t, J = 7.0Hz, 2H). ESI-MS m / z=334.0[M+H] + ;Calcu.=333.1.

[0317] Step E: At 20°C, manganese dioxide (3.0 g, 34.51 mmol) was slowly added to dichloromethane (5 mL) and methanol (5 mL) in which 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(hydroxymethyl)phenyl)ethane-1-ol (500 mg, 1.50 mmol) was dissolved, and the whole mixture was stirred at 20°C for 7 hours. Manganese dioxide (3.0 g, 34.51 mmol) was added to the mixture, and the whole mixture was stirred at 20°C for 15 hours. The reaction solution was filtered through diatomaceous earth, and the filter cake was rinsed with methanol three times (20 mL*3), and the filtrate was collected and directly concentrated under reduced pressure to obtain a solid residue. The solid residue was separated by silica gel column (SiO2, DCM:MeOH=90:10). The reaction afforded a white solid product, 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(2-hydroxyethyl)benzaldehyde (215 mg, 43%). 1 H NMR(500MHz,DMSO-d6)δppm9.92(s,1H),8.42(br s,1H),8.18(br s,1H),8.16(s,1H),7.69(brs,1H),7.45-7.46(m,2H),5.52(s,2H),3.61(m,2H),2.78(t,J=6.2Hz,2H). ESI-MS m / z=332.0[M+H]+; Calcu.=331.1.

[0318] Step F: At 0-5°C, p-toluenesulfonyl chloride (172 mg, 0.904 mmol) was added to dichloromethane (5 mL) in which 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(2-hydroxyethyl)benzaldehyde (200 mg, 0.603 mmol), triethylamine (0.25 mL, 1.809 mmol) and 4-dimethylaminopyridine (36.8 mg, 0.301 mmol) were dissolved, and the whole mixture was stirred at 20°C for 5 hours. The reaction solution was added with water (20 mL), and then extracted twice with dichloromethane (40 mL*2). The organic phases were combined, washed once with saturated sodium chloride aqueous solution (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a solid residue. The solid residue was separated by silica gel column (SiO2, DCM:MeOH=92:8). The reaction gave a pale yellow solid product, 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-formylphenethyl-4-methylbenzenesulfonate (50 mg, 17%). ESI-MS m / z=485.9 [M+H] + ;Calcu.=485.1.

[0319] Step G: At 20°C, cesium carbonate (188 mg, 0.576 mmol) in N,N-dimethylformamide (5 mL) was added to 5-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-2(1H)-one (207 mg, 0.864 mmol) and 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-formylphenethyl-4-methylbenzenesulfonate (140 mg, 0.288 mmol) at N,N-dimethylformamide (5 mL), and the whole mixture was stirred at 20°C for 16 hours. The reaction solution was directly concentrated under reduced pressure to obtain a solid residue. The solid residue was separated by silica gel column (SiO2, DCM:MeOH=90:10). The reaction gave a white solid product, 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(2-(5-(((tert-butyldimethylsilyl)oxy)methyl)-2-oxopyridin-1(2H)-yl)ethyl)benzaldehyde (36 mg, 23%). ESI-MS m / z=553.1[M+H] + ;Calcu.=552.2.

[0320] Step H: At 0-5°C, tetrabutylammonium fluoride (0.046 mL, 0.046 mmol) was slowly added dropwise to tetrahydrofuran (5 mL) in which 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(2-(5-(((tert-butyldimethylsilyl)oxy)methyl)-2-oxopyridin-1(2H)-yl)ethyl)benzaldehyde (17 mg, 0.031 mmol) was dissolved. After the addition was complete, the whole mixture was stirred at 20°C for 2 hours. Water (10 mL) was added to the reaction solution, and the mixture was extracted twice with ethyl acetate (20 mL*2). The organic phases were combined, washed once with a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a light yellow solid product. The crude compound was used directly in the next step without purification. The reaction yielded a pale yellow solid crude product, 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(2-(5-(hydroxymethyl)-2-oxopyridin-1(2H)-yl)ethyl)benzaldehyde (25 mg, crude product). ESI-MS m / z=438.9[M+H] + ;Calcu.=438.1.

[0321] Step I: At 20°C, sodium triacetoxyborohydride (18 mg, 0.085 mmol) was added to dichloromethane (5 mL) in which 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(2-(5-(hydroxymethyl)-2-oxopyridin-1(2H)-yl)ethyl)benzaldehyde (25 mg, crude product) and pyrrole (0.007 mL, 0.085 mmol) were dissolved, and the whole mixture was stirred at 20°C for 18 hours. The reaction solution was directly concentrated under reduced pressure to obtain a solid residue. The solid residue was separated by silica gel column (SiO2, DCM:MeOH=90:10). The reaction gave a yellow oily product, 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(pyrrolidin-1-ylmethyl)phenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one (28 mg, crude product). ESI-MS m / z=494.0 [M+H] + ;Calcu.=493.2.

[0322] Step J: To acetonitrile (20 mL) in which 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(pyrrolidin-1-ylmethyl)phenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one (28 mg, crude) was dissolved was added cesium carbonate (185 mg, 0.57 mmol) and lithium iodide (37.9 mg, 0.28 mmol) at 20°C, and the whole mixture was stirred at 110°C for 2 hours. The reaction solution was filtered through diatomaceous earth, and the filter cake was rinsed with ethyl acetate three times (30 mL*3). The filtrate was collected and concentrated under reduced pressure to obtain a solid residue, which was subjected to prep-HPLC preparation and separation. The preparation conditions were: YMC-ActusTriart C18 (20 mm×250 mm, 5 μm); 5 mM ammonia in water; B: MeCN; 18 mL / min; 0-3 min, 5% B; 3-3.5 min, 5%-15% B; 3.5-15 min, 15%-60% B; detector, UV 254 nm. The reaction obtained compound C-13, a white solid product (E)-14-amino-3 5 -(pyrrolidin-1-ylmethyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (0.843 mg, 3%). ESI-MS m / z=458.1[M+H] + ;Calcu.=457.2.

[0323] Example 12: Synthesis of Compound C-14

[0324]

[0325] Step A: At room temperature, sulfuric acid (0.05 mL, 0.83 mmol) was added to methanol (17 mL) in which (2-methoxy-5-methylphenyl)acetic acid (1.57 g, 8.71 mmol) was dissolved. After the addition was complete, the entire mixture was heated to 80°C and stirred for 4 hours. After TLC monitoring of the disappearance of the raw material, triethylamine solution was added to the system to adjust the pH to neutral, and most of the methanol was evaporated under reduced pressure. Water was added to the concentrate, extracted 3 times with dichloromethane, dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The reaction obtained the product (2-methoxy-5-methylphenyl)acetic acid methyl ester (1.64 g, 97%) as a yellow oil.

[0326] Step B: At 100°C, add azobisisobutyronitrile (0.14g, 0.84mmol) and N-bromosuccinimide (1.81g, 10.13mmol) to 1,2-dichloroethane (20mL) solvent in which (2-methoxy-5-methylphenyl)acetic acid methyl ester (1.64g, 8.44mmol) is dissolved. After stirring evenly, the reaction mixture is refluxed and stirred at 100°C for 7 hours. After the reaction system is cooled to room temperature, 20mL of water is added to the system. The mixed solution is extracted with dichloromethane, the organic phases are combined, washed with saturated brine, dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The reaction obtains the product 2-(5-(bromomethyl)-2-methoxyphenyl)acetic acid methyl ester (2.37g, crude product) as a dark brown liquid.

[0327] Step C: At room temperature, 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.15 g, 6.77 mmol) and cesium carbonate (4.13 g, 112.7 mmol) were added to N,N-dimethylformamide (190 mL) in which methyl 2-(5-(bromomethyl)-2-methoxyphenyl)acetate (2.31 g, 8.46 mmol) was dissolved. After stirring for 4 hours, the raw material disappeared by HPLC detection, the insoluble matter was filtered, the pH of the filtrate was adjusted to neutral with acetic acid, and it was concentrated under reduced pressure. The residue was directly subjected to HPLC preparative separation. The preparation conditions were: Welch Xtimate C18 (30mm×250mm, 5μm); A: 5mM NH4HCO3 in water; B: MeCN; 40mL / min, 0~3min, 10% B; 3~3.5min, 10%~20% B; 3.5~16min, 20%~90% B; detector, UV 254nm. The product 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenyl)acetate (409mg, two-step yield 13%) was obtained as a brown solid. ESI-MS m / z=361.9[M+H] + ;Calcu.=361.1.

[0328] Step D: At room temperature, lithium aluminum hydride (88.0 mg, 2.32 mmol) was added to tetrahydrofuran (40 mL) in which methyl 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenyl)acetate (409 mg, 1.13 mmol) was dissolved. After the addition, the whole mixture was stirred at room temperature for 2 h. TLC detected that the starting material disappeared, water was added to the reaction system to quench the reaction, and dilute hydrochloric acid was added to adjust the pH to neutral. After adding ethyl acetate to the mixture, the filtrate was separated, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The reaction gave the product 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenyl)ethane-1-ol (313 mg, 83%) as a brown solid.

[0329] Step E: At room temperature, triethylamine (0.52 mL, 3.75 mmol), 4-dimethylaminopyridine (11.5 mg, 0.094 mmol) and p-toluenesulfonyl chloride (357 mg, 1.88 mmol) were added to a solution of 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenyl)ethane-1-ol (313 mg, 0.938 mmol) in dichloromethane (35 mL). After the addition, the mixture was stirred at room temperature overnight. The raw material disappeared after TLC detection, the reaction solution was concentrated under reduced pressure, and the residue was separated by silica gel column (DCM: MeOH = 95: 5). The reaction gave a yellow solid product 5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenethyl-4-methylbenzenesulfonate (311 mg, 68%). ESI-MS m / z = 488.0 [M+H] + ;Calcu.=487.1.

[0330] Step F: To N,N-dimethylformamide (30 mL) in which 5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenethyl-4-methylbenzenesulfonate (300 mg, 0.615 mmol) was dissolved, 5-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-2(1H)-one (294 mg, 1.23 mmol) and cesium carbonate (500 mg, 1.54 mmol) were added in sequence at room temperature, and the mixture was stirred for 5 days. The reaction was monitored by HPLC. After the starting material disappeared, the reaction solution was filtered, acetic acid was added to adjust the pH to neutral, N,N-dimethylformamide was evaporated under reduced pressure, and the residue was separated by silica gel column (DCM:MeOH=95:5). The reaction gave a white solid product 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenethyl)-5-(((tert-butyldimethylsilyl)oxy)methyl ester)pyridin-2(1H)-one (156 mg, 46%). ESI-MS m / z=555.1[M+H] + ;Calcu.=554.2.

[0331] Step G: To a solution of 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenethyl)-5-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-2(1H)-one (156 mg, 0.281 mmol) dissolved in tetrahydrofuran (8 ml) at room temperature was added 1 M tetrabutylammonium fluoride solution in tetrahydrofuran (0.422 ml, 0.422 mmol). The reaction mixture was stirred at room temperature for 1.5 hours and monitored by TLC. After the starting material disappeared, tetrahydrofuran was evaporated under reduced pressure, and the residue was separated by silica gel column (DCM: MeOH = 95:5). The reaction gave a white solid product 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one (121 mg, 97%). ESI-MS m / z = 440.9 [M+H] + ;Calcu.=440.1.

[0332] Step H: 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one (115 mg, 0.261 mmol) was dissolved in acetonitrile (30 ml), and cesium carbonate (850 mg, 2.61 mmol) and lithium iodide (350 mg, 2.61 mmol) were added. The reaction solution was heated in a sealed tube at 110°C for 10 hours and monitored by HPLC. After the raw material disappears, the reaction solution is cooled, and the solvent is evaporated under reduced pressure at 35°C. The residue is directly subjected to HPLC preparation and separation. Preparation conditions: Welch Xtimate C18 (30mm×250mm, 5μm); A: 0.05% NH3 in water; B: MeCN; 40mL / min, 0~3min, 5% B; 3~3.5min, 5%~15% B; 3.5~15min, 15%~70% B; detector, UV254nm. The reaction yields compound C-14 and white solid product (E)-1 4 -Amino-3 4 -Methoxy-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (29.99 mg, 28%). 1 H NMR(500MHz,DMSO-d6)δppm 8.30(d,J=2.2Hz,1H),7.93(s,1H),7.68(d,J=1.6Hz,2H),7.42(s,1H),7.36(dd,J=9.3,2.4Hz,1H),7.06(dd,J=8.4,1.8Hz,1 H), 6.80 (d, J = 8.5Hz, 1H), 6.26 (d, J = 9.3Hz, 1H), 5.36 (s, 2H), 5.10 (s, 2H), 4.25-4.40 (m, 2H), 3.66 (s, 3H), 3.03-3.18 (m, 2H). ESI-MS m / z=405.2[M+H] + ;Calcu.=404.2.

[0333] Example 13: Synthesis of Compound C-15

[0334]

[0335] Step A: At room temperature, N-bromosuccinimide (28.9 g, 162.1 mmol) was added to carbon tetrachloride (300 mL) in which 5-bromo-1,3-dimethylbenzene (30 g, 162.1 mmol) and azobisisobutyronitrile (2.7 g, 16.21 mmol) were dissolved. After the addition was complete, the entire mixture was stirred at 90°C for 3 hours. The reaction solution was filtered through diatomaceous earth and concentrated under reduced pressure to obtain a yellow crude product. The crude product was separated by silica gel column (SiO2, PE:EA=95:5), and the reaction obtained a colorless oily product 5-bromo-3-(bromomethyl)-1-methylbenzene (27.4 g, 64%).

[0336] Step B: At room temperature, trimethylsilyl cyanide (20.60 g, 207.6 mmol) was added to acetonitrile (300 mL) in which 5-bromo-3-(bromomethyl)-1-methylbenzene (27.4 g, 103.8 mmol) and potassium carbonate (43.0 g, 311.4 mmol) were dissolved. After the addition was complete, the entire mixture was heated to 60°C and stirred overnight. The reaction solution was filtered and concentrated under reduced pressure to obtain a yellow oily crude product. The crude product was separated by silica gel column (SiO2, PE:EA=95:5), and the reaction obtained a colorless oily product (3-bromo-5-methylphenyl)acetonitrile (18.4 g, 84%).

[0337] Step C: At room temperature, potassium hydroxide (14.74 g, 262.8 mmol) was added to water (50 mL) and ethanol (100 mL) in which (3-bromo-5-methylphenyl)acetonitrile (18.4 g, 87.59 mmol) was dissolved. After the addition was complete, the mixture was heated to 90°C and stirred overnight. The reaction solution was cooled to room temperature, the ethanol was evaporated, water (50 mL) was added, and then dilute hydrochloric acid was added dropwise to adjust the pH value to 1-2. A large amount of white solid was precipitated, and the white solid product (3-bromo-5-methylphenyl)acetic acid (19.2 g, 96%) was obtained by suction filtration.

[0338] Step D: At room temperature, N-bromosuccinimide (14.95 g, 83.99 mmol) was added to carbon tetrachloride (300 mL) in which (3-bromo-5-methylphenyl)acetic acid (19.24 g, 83.99 mmol) and azobisisobutyronitrile (1.38 g, 8.399 mmol) were dissolved. After the addition was complete, the entire mixture was heated to 80°C and stirred overnight. The reaction solution was filtered and concentrated under reduced pressure to obtain a pale yellow oily crude product 2-(3-bromo-5-(bromomethyl)phenyl)acetic acid.

[0339] Step E: At 0°C and under N2 protection, 1M borane tetrahydrofuran solution (14 g, 168.0 mmol) was added to tetrahydrofuran (200 mL) in which the crude 2-(3-bromo-5-(bromomethyl)phenyl)acetic acid from the previous step was dissolved. After the addition was complete, the entire mixture was stirred at room temperature and under nitrogen protection for 3 hours. The reaction solution was temperature controlled at 0°C, and then methanol was slowly added to quench the reaction until no obvious bubbles appeared in the reaction solution. The temperature was controlled not to exceed 10°C during quenching, and then filtered through diatomaceous earth and concentrated under reduced pressure to obtain a yellow crude product. The crude product was separated by silica gel column (SiO2, PE:EA=90:10), and the reaction obtained a white solid product 2-(3-bromo-5-(bromomethyl)phenyl)ethane-1-ol (8.7 g, two-step yield 35%).

[0340] Step F: At room temperature, 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (4.6 g, 27.07 mmol) was added to N,N-dimethylformamide (140 mL) in which 2-(3-bromo-5-(bromomethyl)phenyl)ethane-1-ol (8.7 g, 29.59 mmol) and cesium carbonate (13.2 g, 40.61 mmol) were dissolved. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was filtered and concentrated under reduced pressure to obtain a yellow oily crude product. The crude product was separated by silica gel column (SiO2, DCM:MeOH=95:5) and the reaction obtained a light yellow solid product 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromophenyl)ethane-1-ol (3 g, 27%). ESI-MS m / z=383.9[M+H] + ;Calcu.=383.0.

[0341] Step G: At room temperature, triethylamine (4.36 mL, 31.36 mmol) and 4-dimethylaminopyridine (60 mL) were added to a solution of 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromophenyl)ethane-1-ol (3 g, 7.840 mmol) and p-toluenesulfonyl chloride (2.99 g, 15.68 mmol) in dichloromethane (100 mL). g, 0.523mmol) was added, and the whole mixture was stirred at room temperature for 2 hours, and the reaction solution was concentrated under reduced pressure to obtain a yellow crude product. The crude product was separated by silica gel column (SiO2, DCM: MeOH = 95: 5). The reaction obtained a light yellow solid product 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromophenethyl-4-methylbenzenesulfonate (2.3g, 55%). ESI-MS m / z = 537.8 [M+H] + ;Calcu.=537.0.

[0342] Step H: At room temperature, cesium carbonate (2.09 g, 6.427 mmol) was added to N,N-dimethylformamide (20 mL) in which 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromophenethyl-4-methylbenzenesulfonate (2.3 g, 4.284 mmol) and 6-oxo-1,6-dihydropyridine-3-carbaldehyde (0.69 g, 5.57 mmol) were dissolved. After the addition was complete, the whole mixture was stirred at room temperature overnight. Subsequently, the product was concentrated under reduced pressure to obtain a yellow crude product. The residue was dissolved in N,N-dimethylformamide (15 mL) and subjected to reverse phase separation. Chromatographic column: X select C18 (19 mm x 150 mm); mobile phase: water (containing 0.1% formic acid) and acetonitrile; flow rate: 25 ml / min; gradient: acetonitrile increased from 0% to 100% in 30 minutes; detection wavelength: 254 nm. The product was freeze-dried under reduced pressure to obtain a light yellow solid product 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromophenethyl)-6-oxo-1,6-dihydropyridine-3-carboxaldehyde (400 mg, 19%). ESI-MS m / z=488.8[M+H] + ;Calcu.=488.0.

[0343] Step I: At 0°C, sodium borohydride (77.6 mg, 2.05 mmol) was added to tetrahydrofuran (40 mL) in which 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromophenethyl)-6-oxo-1,6-dihydropyridine-3-carbaldehyde (400 mg, 0.82 mmol) was dissolved. After the addition was complete, the entire mixture was stirred at room temperature for 2 hours. Subsequently, the mixture was concentrated under reduced pressure to obtain a yellow crude product, 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromophenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one. ESI-MS m / z=490.8[M+H] + ;Calcu.=490.0.

[0344] Step J: At room temperature, lithium iodide (1.1 g, 8.201 mmol) was added to acetonitrile (60 mL) in which the crude product 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromophenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one and cesium carbonate (2.7 g, 8.201 mmol) were dissolved. After the addition was complete, the whole mixture was heated to 110°C and stirred for 16 hours, and then concentrated under reduced pressure to obtain a yellow crude product. The crude product was subjected to HPLC preparation and separation, preparation conditions: Waters XBridge C18 (10 mm×150 mm, 5 μm); A: 5 mM NH3 in water, B: MeCN; 5 mL / min; 0-3 min, 5% B; 3-15 min, 5%-50% B; detector, UV 254 nm). The reaction yielded compound C-15, a white solid product (E)-1 4 -Amino-3 5 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (300 mg, two-step yield 81%). 1 H NMR(DMSO-d6,500MHz)δ8.08(d,J=2.3Hz,1H),7.98(s,1H),7.71(s,2H),7.51(br s,1H),7.33(dd,J=9.3,2.4Hz,1H),7.18(s,1H),6.96(s,1H),6.25(d,J=9.3H z, 1H), 5.43 (s, 2H), 5.04 (s, 2H), 4.30 (t, J = 5.9Hz, 2H), 3.20 (t, J = 5.7Hz, 2H). ESI-MS m / z=453.2[M+H] + ;Calcu.=452.1.

[0345] Example 14: Synthesis of Compound C-16

[0346]

[0347] Step A: At room temperature, dissolve (E)-1 4 -Amino-3 5 -Br-6 1 ,6 2 -Dihydro-1 1H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (50mg, 0.110mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (10.5mg, 0.022mmol), sodium carbonate (36.3mg, 0.343mmol) in water (1mL) and dioxane (4mL), tris(dibenzylideneacetone)dipalladium (5.1mg, 0.006mmol) was added, and after the addition, the whole mixture was heated to 80℃ and stirred for 3 hours under nitrogen protection. Then it was concentrated under reduced pressure to obtain a yellow crude product. The crude product was subjected to HPLC preparation and separation, preparation conditions: Waters XBridge C18 (10mm×150mm, 5μm); A: 5mM NH3 inwater, B: MeCN; 5mL / min; 0~3min, 5% B; 3~15min, 5%~50% B; detector, UV 254nm). The reaction yielded compound C-16, a white solid product (E)-1 4 -Amino-3 5 -((dimethylamino)methyl)-6- 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (3.12 mg, 7%). 1 H NMR(DMSO-d6,500MHz)δppm 8.04(d,J=2.1Hz,1H),7.95(s,1H),7.65-7.77(m,1H),7.52(s,2H),7.31(dd,J=9.3,2.4Hz,1H),6.82(s,1H),6.76(s, 1H), 6.23 (d, J = 9.3Hz, 1H), 5.41 (s, 2H), 5.03 (s, 2H), 4.26-4.31 (m, 2H), 3.15-3.18 (m, 2H), 3.13 (s, 2H), 1.97 (s, 6H). ESI-MS m / z=432.3[M+H] + ;Calcu.=431.2.

[0348] Example 15: Synthesis of Compound C-17

[0349]

[0350] Step A: At room temperature and under nitrogen protection, dissolve (E)-1 4 -Amino-34 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (60mg, 0.132mmol) in 1,4-dioxane (10mL), dimethylphosphine oxide (31mg, 0.397mmol), tris(dibenzylideneacetone)dipalladium (60mg, 0.066mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (63mg, 0.109mmol) and potassium phosphate (28mg, 0.396mmol) were added. After the addition, the whole mixture was heated to 100°C and stirred for 18 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (30mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 40mL / min; 0-3min, 5% B; 3-3.5min, 5%-5% B; 3.5-15min, 5%-50% B; 15-15.5min, 50%-100% B; detector, UV 254nm. Compound C-17 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -(Dimethylphosphoryl)-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (15.06 mg, 25%). 1 H NMR (500MHz, DMSO-d6) δppm 8.25 (d, J = 2.0Hz, 1H), 7.97 (s, 1H), 7.86 (s, 1H), 7.73 (br s,1H),7.42-7.49(m,2H),7.34(dd,J=9.2,2.5Hz,1H),7.02(d,J=7.8Hz,1H),6.26(d,J=9.2Hz,1H) ,5.49(s,2H),5.05(s,2H),4.42(t,J=5.3Hz,2H),3.58(t,J=5.3Hz,2H),1.61(s,3H),1.58(s,3H). ESI-MS m / z=451.3[M+H] + ;Calcu.=450.2.

[0351] Example 16: Synthesis of Compound C-18 / M-1

[0352]

[0353] Step A: At room temperature, thionyl chloride (1.7 mL, 24.0 mmol) was added to methanol (30 mL) in which 5-methyl-6-oxo-1,6-dihydropyridine-3-carboxylic acid (919 mg, 6.0 mmol) was dissolved. After the addition, the whole mixture was stirred at room temperature and monitored by HPLC until the reaction of the raw material was complete. The mixture was concentrated to dryness under reduced pressure. The residue was repeatedly diluted with dichloromethane and concentrated to dryness under reduced pressure. About 5 mL of dichloromethane was added, the mixture was filtered, and the filter cake was washed with a small amount of dichloromethane and dried. The reaction gave a brown solid product, 5-methyl-6-oxo-1,6-dihydropyridine-3-carboxylic acid methyl ester (660 mg, 66%). ESI-MS m / z=168.0[M+H] + ;Calcu.=167.1.

[0354] Step B: At room temperature, cesium carbonate (977 mg, 3.0 mmol) was added to N,N-dimethylformamide (30 mL) in which 5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl-4-methylbenzenesulfonate (805 mg, 1.5 mmol) and 5-methyl-6-oxo-1,6-dihydropyridine-3-carboxylic acid methyl ester (501 mg, 3.0 mmol) were dissolved. After the addition, the whole mixture was stirred at room temperature for 48 hours, filtered and concentrated under reduced pressure to obtain a solid residue, which was separated by silica gel column (SiO2, DCM:MeOH=94:6). The reaction obtained the product 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-5-methyl-6-oxo-1,6-dihydropyridine-3-carboxylic acid methyl ester (340 mg, 43%) as a light yellow solid. ESI-MSm / z=532.8[M+H] + ;Calcu.=532.0.

[0355] Step C: In an ice bath and nitrogen protection, diisobutylaluminum hydride (3.2 mL, 3.2 mmol) was slowly added to dichloromethane (32 mL) in which 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-5-methyl-6-oxo-1,6-dihydropyridine-3-carboxylic acid methyl ester (340 mg, 0.64 mmol) was dissolved. After the addition was complete, the entire mixture was stirred in an ice bath for 1 hour. About 5 mL of methanol was added to quench the reaction, and the mixture was concentrated to dryness under reduced pressure. The residue was separated by column chromatography (SiO2, DCM:MeOH=93:7). The reaction gave a white solid product, 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-5-(hydroxymethyl)-3-methylpyridin-2(1H)-one (143 mg, 44%). ESI-MS m / z=486.8[M-OH] + ;Calcu.=504.0.

[0356] Step D: At room temperature, 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-5-(hydroxymethyl)-3-methylpyridin-2(1H)-one (120 mg, 0.24 mmol) was dissolved in acetonitrile (30 mL), and cesium carbonate (780 mg, 2.4 mmol) and lithium iodide (324 mg, 2.4 mmol) were added. After the addition, the whole mixture was heated to 110°C and stirred for 7 hours. After filtration, the solid residue was concentrated under reduced pressure and separated by silica gel column (SiO2, EA:MeOH=90:10). The reaction gave M-1 and a light yellow solid product (E)-1 4 -Amino-3 4 -Br-6 3 -Methyl-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (52 mg, 47%). ESI-MS m / z=467.2[M+H] + ;Calcu.=466.1.

[0357] Step E: At room temperature and under N2 protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 3 -Methyl-6 1 ,6 2 -Dihydro-1 1H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (26 mg, 0.05 mmol), 1-((trifluoro-4-boryl)methyl)pyrrolidine potassium salt (28.6 mg, 0.15 mmol), sodium carbonate (15.9 mg, 0.15 mmol) and 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (9.5 mg, 0.02 mmol) in 1,4-dioxane (1.6 mL) and water (0.4 mL) were added tris(dibenzylideneacetone)dipalladium (9.1 mg, 0.01 mmol). After the addition was complete, the whole mixture was heated to 90 ° C and stirred under nitrogen for 5 hours. Filter by suction, wash with a small amount of methanol, concentrate to dryness under reduced pressure, and perform HPLC separation on the residue (preparation conditions: YMC-Actus Triart C18 (21mm×250mm, 5μm); A: 0.05% ammonia water, B: MeCN; 18mL / min; 0~3min, 5% B; 3~18min, 5%~60% B; detector, UV 254nm). The reaction yields compound C-18 and white solid product (E)-1 4 -Amino-6 3 -Methyl-3 4 -(pyrrolidin-1-ylmethyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (4 mg, 15%). 1 H NMR(500MHz,DMSO-d6)δppm 8.11(d,J=1.1Hz,1H),7.93(s,1H),7.69(brs,1H),7.62(s,1H),7.69(br s,1H),7.25(s,1H),7.10(d,J=7.8Hz,1H),6.96(d,J=7.9Hz,1H),5.40(s,2H),5.05(s,2H),4 .34(t,J=5.0Hz,2H),3.40(s,2H),3.24-3.27(m,2H),2.33(m,4H),1.89(s,3H),1.64(m,4H). ESI-MS m / z=472.3[M+H] + ;Calcu.=471.2.

[0358] Example 17: Synthesis of Compound C-19

[0359]

[0360] Step A: At room temperature and under N2 protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (20mg, 0.044mmol) in 2-methyltetrahydrofuran (0.5mL), methanesulfonamide (7mg, 0.074mmol), allylpalladium (II) chloride dimer (2mg, 0.005mmol), 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (7mg, 0.016mmol) and potassium carbonate (12mg, 0.087mmol) were added. After the addition, the whole mixture was heated to 80°C and stirred for 18 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridgeC18 (19mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 18mL / min; 0-3min, 5% B; 3-3.5min, 5%-10% B; 3.5-15min, 10%-50% B; 15-15.5min, 50%-100% B; detector, UV 254nm. Compound C-19 was obtained, and a white solid product (E)-N-(1 4 -Amino-6 2 -oxy-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzoic acid cyclooctane-3 4 -yl)methanesulfonamide (2.48 mg, 8%).

[0361] 1H NMR(500MHz,DMSO-d6)δppm 8.28(d,J=2.3Hz,1H),7.94(s,1H),7.75(s,1H),7.72(br s,1H),7.46(br s,1H),7.36(dd,J=9.4,2.3Hz,1H),7.21(br s,1H),7.12(d,J=8.4Hz,1H),6.99(d,J=8.4Hz,1H),6.25(d,J=9.4Hz,1H),5.41 (s, 2H), 5.07 (s, 2H), 4.34 (t, J = 5.3Hz, 2H), 3.26 (t, J = 5.3Hz, 2H), 2.83 (s, 3H). ESI-MS m / z=468.2[M+H] + ;Calcu.=467.1.

[0362] Example 18: Synthesis of Compound C-20

[0363]

[0364] Step A: At room temperature, dissolve (E)-1 4 -Amino-3 5 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (50mg, 0.11mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (10.5mg, 0.022mmol), sodium carbonate (36.3mg, 0.343mmol) in water (1mL) and dioxane (4mL), tris(dibenzylideneacetone)dipalladium (5.1mg, 0.006mmol) was added, and after the addition, the whole mixture was heated to 80℃ and stirred for 3 hours under nitrogen protection. Then it was concentrated under reduced pressure to obtain a yellow crude product. The crude product was subjected to HPLC preparation and separation, preparation conditions: Waters XBridge C18 (10mm×150mm, 5μm); A: 5mM NH3 inwater, B: MeCN; 5mL / min; 0~3min, 5% B; 3~15min, 5%~50% B; detector, UV 254nm). The reaction gave compound C-20, white solid product (E)-1 4 -Amino-3 5 -(Morpholinemethyl)-6 1 ,6 2 -Dihydro-1 1H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (2.93 mg, 6%). 1 HNMR(DMSO-d6,500MHz)δppm 7.97(s,1H),7.93(d,J=2.3Hz,1H),7.71(br s,1H),7.52(s,2H),7.28(dd,J=9.3,2.4Hz,1H),6.76(s,1H),6.57(s,1H),6.22(d,J=9.3Hz,1H),5.42(s,2H),4.99(s,2H),4.27(br t,J=5.8Hz,2H),3.43(br t,J=4.2Hz,4H),3.12-3.19(m,4H),2.04(br s,4H). ESI-MS m / z=474.1[M+H] + ;Calcu.=473.2.

[0365] Example 19: Synthesis of Compound C-21

[0366]

[0367] Step A: At room temperature and under nitrogen protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2-ketone (20 mg, 0.044 mmol) in dioxane (1 mL), dimethylphosphine oxide (10 mg, 0.128 mmol), tris(dibenzylideneacetone)dipalladium (4 mg, 0.004 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (5 mg, 0.009 mmol) and potassium phosphate (28 mg, 0.132 mmol) were added. After the addition was complete, the whole mixture was heated to 100 ° C and stirred for 18 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (30mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 40mL / min; 0-3min, 5% B; 3-3.5min, 5%-10% B; 3.5-12min, 10%-60% B; 12-15min, 60%-100% B; detector, UV254nm. Compound C-21 and white solid product (E)-1 were obtained. 4 -Amino-3 5 -(Dimethylphosphoryl)-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (12.53 mg, 63%).

[0368] 1 H NMR (500MHz, DMSO-d6) δppm 8.13 (d, J=1.6Hz, 1H), 7.97 (s, 1H), 7.82 (s, 1H), 7.75 (br s, 1H), 7.49 (br s,1H),7.32-7.41(m,3H),6.26(d,J=9.5Hz,1H),5.48(s,2H),5.07(s,2H ), 4.32 (t, J = 5.4Hz, 2H), 3.26 (t, J = 5.4Hz, 2H), 1.51 (s, 3H), 1.49 (s, 3H). ESI-MS m / z=451.3[M+H] + ;Calcu.=450.2.

[0369] Example 20: Synthesis of Compound C-22 / C-30

[0370]

[0371] Step A: At room temperature and under nitrogen protection, dissolve (E)-14 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (60mg, 0.132mmol) in dioxane (10mL), tributyl (1-ethoxyethylene) tin (143mg, 0.396mmol) and 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (97mg, 0.132mmol) were added. After the addition, the whole mixture was heated to 85°C and stirred for 18 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (30mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 40mL / min; 0~3min, 5% B; 3~3.5min, 5%~15% B; 3.5~11min, 15%~60% B; 11~15min, 60%~100% B; detector, UV 254nm. Obtained compound C-30, white solid product (E)-1 4 -Amino-3 4 -(1-ethoxyvinyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (16.47 mg, 28%). 1H NMR (500MHz, DMSO-d6) δppm 8.23 ​​(d, J=2.3Hz, 1H), 7.95 (s, 1H), 7.73 (s, 1H), 7.71 (br s, 1H), 7.46 (br s,1H),7.36(dd,J=9.2,2.3Hz,1H),7.07(d,J=7.8Hz,1H),6.96(d,J=7.8Hz,1H),6.26(d,J=9.2Hz,1H),5.45(s,2H),5.06(s,2H),4. 32(t,J=5.0Hz,2H), 4.28(d,J=1.8Hz,1H), 3.99(d,J=1.8Hz,1H), 3.78(q,J=6.8Hz,2H), 3.22(t,J=5.0Hz,2H), 1.25(t,J=6.8Hz,3H). ESI-MS m / z=445.3[M+H] + ;Calcu.=444.2.

[0372] Step B: At room temperature, dissolve (E)-1 4 -Amino-3 4 -(1-ethoxyvinyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (14.81 mg, 0.033 mmol) in acetonitrile (containing 1% formic acid, 30 mL), the whole mixture was stirred at room temperature for 0.5 hours. The reaction solution was concentrated under reduced pressure to obtain a white solid crude residue, and the residue was subjected to HPLC preparation and separation. Preparation conditions: YMC-Actus TriartC18 (21mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 18mL / min; 0~3min, 10% B; 3~3.5min, 10%~20% B; 3.5~15min, 20%~80% B; 15~15.5min, 80%~100% B; detector, UV 254nm. Compound C-22 and white solid product (E)-3 were obtained. 4 -Acetyl-1 4 -Amino-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2-ketone (0.44 mg, 3%). ESI-MS m / z=417.2[M+H] + ;Calcu.=416.2.

[0373] Example 21: Synthesis of Compound C-23 / M-2

[0374]

[0375] Step A: At room temperature and under nitrogen protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (20mg, 0.044mmol) in 1,4-dioxane (5mL), tributyl (1-ethoxyethylene) tin (48mg, 0.132mmol) and 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (32mg, 0.044mmol) were added. After the addition, the whole mixture was heated to 85°C and stirred for 18 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (30mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 40mL / min; 0~3min, 5% B; 3~3.5min, 5%~15% B; 3.5~11min, 15%~60% B; 11~15min, 60%~100% B; detector, UV 254nm. M-2, white solid product (E)-1 was obtained 4 -Amino-3 5 -(1-ethoxyvinyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (2.45 mg, 12%). ESI-MS m / z=445.3[M+H] + ;Calcu.=444.2.

[0376] Step B: At room temperature, dissolve (E)-1 4 -Amino-3 5 -(1-ethoxyvinyl)-61 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (2.45mg, 0.006mmol) in acetonitrile (containing 1% formic acid, 10mL), the whole mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain a white solid crude residue, and the residue was subjected to HPLC preparation and separation, preparation conditions: Waters XBridge C18 (30mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 40mL / min; 0~3min, 5% B; 3~3.5min, 5%~15% B; 3.5~11min, 15%~60% B; 11~15min, 60%~100% B; detector, UV 254nm. Compound C-23, white solid product (E)-3 5 -Acetyl-1 4 -Amino-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (0.36 mg, 16%). ESI-MS m / z=417.2[M+H] + ;Calcu.=416.2.

[0377] Example 22: Synthesis of Compound C-24

[0378]

[0379] Step A: At room temperature, add (E)-1 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2-Ketone (27 mg, 0.060 mmol), zinc cyanide (5.6 mg, 0.048 mmol), methanesulfonic acid (2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl) (2'-amino-1,1'-biphenyl-2-yl) palladium (II) (24 mg, 0.030 mmol), after addition, the reaction system was replaced with nitrogen three times. Then tetrahydrofuran (1.3 mL) and H2O (1.3 mL) were added, and deoxygenation (degas) was performed three times to remove oxygen. After that, under a nitrogen atmosphere, the entire mixture was heated to 90 ° C and stirred for 3 hours. Post-treatment: methanol (20 mL) was added for ultrasonic dissolution, filtered through a sand core funnel with silica gel and concentrated under reduced pressure. Separation and purification: separated by preparative HPLC. Chromatographic conditions: YMC-Actus Triart C18 (21mm×250mm, 5μm), A: 0.05% ammonia water, B: ACN; 18mL / min; 0~3.0min, 10% B; 3.0~3.5min, 10%~30% B; 3.5~15min, 30%~80% B; 15~15.5min, 80%~100% B; detector, UV 254nm), after freeze-drying, compound C-24 was obtained as a white solid (E)-1 4 -Amino-6 2 -oxy-6 1 ,6 2 -Dihydro-11H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzazepin-3 4 -nitrile (0.43 mg, 1%). 1 H NMR(500MHz,DMSO-d6)δppm 8.20(d,J=2.0Hz,1H),8.01-8.03(m,1H),7.99(s,1H),7.75(br s,1H),7.57(d,J=8.0Hz,1H),7.47(br s,1H),7.34(dd,J=9.0,2.0Hz,1H),6.98(dd,J=8.0,1.0Hz,1H),6.26(d,J=9.0 Hz, 1H), 5.53 (s, 2H), 5.03 (s, 2H), 4.43 (t, J = 6.0Hz, 2H), 3.47 (t, J = 6.0Hz, 2H). ESI-MS m / z=400.2[M+H] + ;Calcu.=399.1.

[0380] Example 23: Synthesis of Compound C-25

[0381]

[0382] Step A: At room temperature, add (E)-1 to the Schlenk reaction tube in sequence. 4 -Amino-3 5 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (10 mg, 0.022 mmol), zinc cyanide (2.0 mg, 0.017 mmol), methanesulfonic acid (2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl) (2'-amino-1,1'-biphenyl-2-yl) palladium (II) (8.0 mg, 0.010 mmol), after addition, the reaction system was replaced with nitrogen three times. Then tetrahydrofuran (0.5 mL) and H2O (0.5 mL) were added, and then degased (degas) was performed three times to remove oxygen. After that, under a nitrogen atmosphere, the entire mixture was heated to 90 ° C and stirred for 3 hours. Post-treatment: methanol (20 mL) was added for ultrasonic dissolution, filtered through a sand core funnel with silica gel, and concentrated under reduced pressure. Separation and purification: separated by preparative HPLC. Chromatographic conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm), A: 0.05% ammonia water, B: ACN; 18 mL / min; 0-3.5 min, 5% B; 3.5-12 min, 5%-50% B; 12-15 min, 50%-80% B detector, UV 254 nm), after freeze-drying, compound C-25 and white solid product (E)-1 were obtained. 4 -Amino-6 2 -oxy-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzazepin-3 5 -nitrile (0.96 mg, yield 10%). 1H NMR (500MHz, DMSO-d6) δppm 8.07 (d, J=2.5Hz, 1H), 8.04 (s, 1H), 8.00 (s, 1H), 7.76 (br s, 1H), 7.53 (br s,1H),7.47(s,1H),7.32(dd,J=9.5,2.5Hz,1H),7.17(s,1H),6.25(d,J=9.5 Hz, 1H), 5.50 (s, 2H), 5.04 (s, 2H), 4.32 (t, J = 5.5Hz, 2H), 3.25-3.27 (m, 2H). ESI-MS m / z=400.0[M+H] + ;Calcu.=399.1.

[0383] Example 24: Synthesis of Compound C-26

[0384]

[0385] Step A: Add (E)-1 into the Schlenk reaction tube at room temperature. 4 -Amino-3 4 -Br-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)benzocyclooctane-6 6 -Ketone (45mg, 0.099mmol), zinc cyanide (18mg, 0.153mmol), methanesulfonic acid (2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl) (2'-amino-1,1'-biphenyl-2-yl) palladium (II) (40mg, 0.050mmol). After the addition, the reaction system was replaced with nitrogen three times. Then, tetrahydrofuran (2.0mL) and water (2.0mL) were added, and then degassing was performed 3 times to remove oxygen. After that, under a nitrogen atmosphere, the entire mixture was heated to 90°C and stirred for 5 hours. Methanol (20mL) was added for ultrasonic dissolution, filtered through a sand core funnel with silica gel, and concentrated under reduced pressure. Separation and purification: separated by preparative HPLC. Chromatographic conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm), A: 0.05% ammonia water, B: ACN; 18 mL / min; 0-3.0 min, 10% B; 3.0-17 min, 10%-80% B; 17-18 min, 80%-100% B detector, UV 254 nm). After freeze-drying, compound C-26 and white solid product (E)-1 were obtained. 4 -Amino-6 6 -oxy-61 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)benzene ring octylamide-3 4 -nitrile (8.85 mg, 21%). 1 H NMR (500MHz, DMSO-d6) δppm7.94(s,1H),7.89(s,1H),7.84(br s,1H),7.68(d,J=7.5Hz,1H),7.61(br s,1H),7.26-7.32(m,2H),6.81(d,J=10.0Hz,1H),5.36(s,2H),5.32(s,2H),4.58-4.61(m,2H),3.49-3.52(m,2H). ESI-MS m / z=401.0[M+H] + ;Calcu.=400.1.

[0386] Example 25: Synthesis of Compound C-27

[0387]

[0388] Step A: At room temperature and under nitrogen protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6-ketone (50mg, 0.11mmol) in dioxane (10mL), dimethylphosphine oxide (26mg, 0.333mmol), tris(dibenzylideneacetone)dipalladium (50mg, 0.055mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (64mg, 0.111mmol) and potassium phosphate (70mg, 0.33mmol) were added. After the addition, the whole mixture was heated to 100°C and stirred for 18 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Welch C18 (30mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 40mL / min; 0-3min, 5% B; 3-3.5min, 5%-10% B; 3.5-15min, 10%-50% B; 15-15.5min, 50%-100% B; detector, UV254nm. Compound C-27 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -(Dimethylphosphoryl)-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -Ketone (18.92 mg, 38%). 1 H NMR (500MHz, DMSO-d6) δppm 7.93 (s, 1H), 7.85 (br s, 1H), 7.69 (d, J = 1.7Hz, 1H), 7.61 (br s,1H),7.52(dd,J=13.2,8.1Hz,1H),7.27(d,J=8.1Hz,1H),7.25(d,J=9.8Hz,1H),6.77(d,J=9.8Hz,1 H), 5.33 (s, 2H), 5.28 (s, 2H), 4.59 (t, J = 5.6Hz, 2H), 3.69 (t, J = 5.6Hz, 2H), 1.67 (s, 3H), 1.64 (s, 3H). ESI-MS m / z=452.2[M+H] + ;Calcu.=451.2.

[0389] Example 26: Synthesis of Compound C-28 / C-29

[0390]

[0391] Step A: At room temperature and under nitrogen protection, dissolve (E)-14 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (20mg, 0.044mmol) in dimethyl sulfoxide (1mL), add cuprous iodide (17mg, 0.089mmol) and triethylenediamine (20mg, 0.178mmol). After the addition, the whole mixture was heated to 145℃ and stirred for 18 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown liquid crude residue, and the residue was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (19mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 18mL / min; 0~3min, 5% B; 3~13min, 5%~40% B; 13~20min, 40%~80% B; 20~21min, 80%~100% B; detector, UV 254nm. C-29, white solid product (E)-1 was obtained 4 -Amino-3 5 -(Methylsulfinyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (0.39 mg, 2%), ESI-MS m / z=437.2 [M+H] + ; Calcu.=436.1. and C-28, white compound (E)-1 4 -Amino-3 5 -(Methylsulfonyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (0.28 mg, 0.001 mmol, 1%), ESI-MS m / z=453.3 [M+H] + ;Calcu.=452.1.

[0392] Example 27: Synthesis of Compound C-31 / C-33

[0393]

[0394] Step A: At room temperature and under nitrogen protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -ketone (100 mg, 0.220 mmol) in 1,4-dioxane (20 mL), tributyl (1-ethoxyethylene) tin (238 mg, 0.659 mmol) and 1,1'-bisdiphenylphosphinoferrocene palladium dichloride (161 mg, 0.220 mmol) were added. After the addition was complete, the whole mixture was heated to 85 ° C and stirred under nitrogen protection for 18 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparation and separation. Preparation conditions: Welch C18 (30mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 40mL / min; 0-3min, 10% B; 3-3.5min, 10%-20% B; 3.5-15min, 20%-85% B; 15-15.5min, 85%-100% B; detector, UV 254nm. Compound C-31 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -(1-ethoxyvinyl)-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -Ketone (26.83 mg, 27%). 1H NMR(500MHz,DMSO-d6)δppm 7.91(s,1H),7.81(br s,1H),7.77(s,1H),7.56(br s,1H),7.35(d,J=9.5Hz,1H),7.19(dd,J=7.8,1.4Hz,1H),7.13(d,J=7.8Hz,1H),6.81(d,J=9.5Hz,1H),5.36(s,2H),5.28(s,2H),4. 49(t,J=5.4Hz,2H), 4.33(d,J=1.9Hz,1H), 4.04(d,J=1.9Hz,1H), 3.82(q,J=7.1Hz,2H), 3.32(t,J=5.4Hz,2H), 1.27(t,J=7.1Hz,3H). ESI-MS m / z=446.3[M+H] + ;Calcu.=445.2.

[0395] Step B: At room temperature, dissolve (E)-1 4 -Amino-3 4 -(1-ethoxyvinyl)-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -ketone (25mg, 0.056mmol) in acetonitrile (containing 1% formic acid, 20mL), the whole mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure to obtain a white solid crude residue, and the residue was subjected to HPLC preparation and separation. The preparation conditions were: YMC-Actus TriartC18 (21mm×250mm, 5μm); A: 0.1% formic acid in water, B: MeCN; 18mL / min; 0~3min, 10% B; 3~3.5min, 10%~15% B; 3.5~15min, 15%~75% B; 15~15.5min, 75%~100% B; detector, UV 254nm. Compound C-33, white solid product (E)-3 4 -Acetyl-1 4 -Amino-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -Ketone (9.21 mg, 39%). 1H NMR (500MHz, DMSO-d6) δppm 7.94 (s, 1H), 7.85 (br s, 1H), 7.69 (d, J = 7.9Hz, 1H), 7.61 (s, 1H), 7.60 (br s,1H),7.21(d,J=7.9Hz,1H),7.13(d,J=9.6Hz,1H),6.68(d,J=9.6Hz,1H),5.31 (s,2H),5.28(s,2H),4.41(t,J=6.1Hz,2H),3.50(t,J=6.1Hz,2H),2.48(s,3H). ESI-MS m / z=418.2[M+H] + ;Calcu.=417.2.

[0396] Example 28: Synthesis of Compound C-32

[0397]

[0398] Step A: At room temperature, add (E)-1 to the Schlenk reaction tube in sequence. 4 -Amino-3 4 -Br-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -Ketone (36 mg, 0.079 mmol), cyclopropylboronic acid (70 mg, 0.815 mmol), methanesulfonic acid (2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl) (2'-amino-1,1'-biphenyl-2-yl) palladium (II) (67 mg, 0.084 mmol), potassium carbonate (77 mg, 0.557 mmol), after addition, the reaction system was replaced with nitrogen three times. Then 1,4-dioxane (24 mL) and H2O (6 mL) were added, and deoxygenation (degas) was performed 3 times to remove oxygen. After that, under a nitrogen atmosphere, the entire mixture was heated to 90 ° C and stirred for 24 hours. After that, methanol was added to dissolve by ultrasound, filtered and concentrated under reduced pressure. After that, the resulting mixture was separated by preparative HPLC. Chromatographic conditions: Welch C18 OBD (30 mm × 250 mm, 5 μm), A: 0.1% FA, B: ACN; 40 mL / min; 0-3.0 min, 10% B; 3.0-15 min, 10%-70% B; 15-15.5 min, 70%-100% B detector, UV 254 nm), after freeze-drying, compound C-32 and white solid product (E)-1 were obtained. 4 -Amino-34 -Cyclopropyl-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -Ketone (4.02 mg, 4%).

[0399] 1 H NMR(500MHz,DMSO-d6)δppm 7.89(s,1H),7.79(br s,1H),7.71(s,1H),7.56(brs,1H),7.31(d,J=10.0Hz,1H),7.12(d,J=8.5Hz,1H),6.88(d,J=8.5Hz,1H),6.80(d,J=10.0Hz,1H), 5.37(s,2H),5.22(s,2H),4.59(t,J=6.0Hz,2H),3.44(t,J=6.0Hz,2H),2.00-2.02(m,1H),0.86-0.88(m,2H),0.49-0.52(m,2H). ESI-MS m / z=416.0[M+H] + ;Calcu.=415.2.

[0400] Example 29: Synthesis of Compound C-34 / C-35 / C-36

[0401]

[0402] Step A: At room temperature and under nitrogen protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6-ketone (20mg, 0.044mmol) in dimethyl sulfoxide (1mL), add cuprous iodide (17mg, 0.089mmol) and triethylenediamine (20mg, 0.178mmol). After the addition, the whole mixture was heated to 145°C and stirred for 18 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown liquid crude residue, and the residue was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (19mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 18mL / min; 0~3min, 30% B; 3~15min, 30%~70% B; 15~16min, 70%~100% B; detector, UV 254nm. Compound C-34 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -(Methylthio)-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -Ketone (2.30 mg, 12.40%). 1 H NMR(500MHz,DMSO-d6)δppm 7.90(s,1H),7.80(br s,1H),7.77(s,1H),7.56(br s,1H),7.35(d,J=9.7Hz,1H),7.27(d,J=8.2Hz,1H),7.15(d,J=8.2Hz,1H),6.82(d,J=9.7H z, 1H), 5.37 (s, 2H), 5.25 (s, 2H), 4.57 (t, J = 5.4Hz, 2H), 3.23 (t, J = 5.4Hz, 2H), 2.40 (s, 3H). ESI-MS m / z=422.0[M+H] + ;Calcu.=421.1.

[0403] Step B: At room temperature, dissolve (E)-1 4 -Amino-3 4 -(Methylthio)-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6-ketone (40mg, 0.095mmol) in dichloromethane (4mL), add meta-chloroperbenzoic acid (100mg, 85% purity, 0.491mmol). After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a white solid crude residue, and the residue was subjected to HPLC preparation and separation. Preparation conditions: Welch C18 (30mm×250mm, 5μm); A: 0.1% formic acid in water, B: MeCN; 40mL / min; 0~3min, 10% B; 3~3.5min, 10%~15% B; 3.5~15min, 15%~60% B; 15~15.5min, 60%~100% B; detector, UV 254nm. Compound C-35, white solid product (E)-1 4 -Amino-3 4 -(Methylsulfinyl)-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -Ketone (6.36 mg, 16%). 1 H NMR (500MHz, DMSO-d6) δppm 7.94 (s, 1H), 7.83 (br s, 1H), 7.76 (s, 1H), 7.73 (d, J = 8.0Hz, 1H), 7.59 (br s,1H),7.42(dd,J=8.0,1.4Hz,1H),7.29(d,J=9.8Hz,1H),6.78(d,J=9.8Hz,1H ), 5.27-5.38 (m, 4H), 4.57 (t, J = 6.4Hz, 2H), 3.32 (t, J = 6.4Hz, 2H), 2.62 (s, 3H). ESI-MS m / z=438.0[M+H] + ; Calcu.=437.1. And compound C-36, white solid compound (E)-1 4 -Amino-3 4 -(Methylsulfonyl)-6 1 ,6 6 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 -Ketone (1.63 mg, 4%). 1H NMR(500MHz,DMSO-d6)δppm 7.95(s,1H),7.80-7.87(m,3H),7.60(br s,1H),7.44(dd,J=8.1,1.4Hz,1H),7.32(d,J=9.6Hz,1H),6.81(d,J=9.6Hz,1H),5 .37(s,2H),5.31(s,2H),4.63(t,J=5.6Hz,2H),3.71(t,J=5.6Hz,2H),3.16(s,3H). ESI-MS m / z=454.0[M+H] + ;Calcu.=453.1.

[0404] Example 30: Synthesis of Compound C-37

[0405]

[0406] Step A: At 0°C, lithium aluminum hydride (2.68 g, 70.66 mmol) was added to tetrahydrofuran (150 mL) in which methyl 3-bromo-4-(trifluoromethyl)benzoate (20 g, 70.66 mmol) was dissolved. After the addition was complete, the entire mixture was stirred at room temperature for 3 hours. The reaction solution was temperature controlled at 0°C, and then the quenching reagent methanol was slowly added until no obvious bubbles appeared in the reaction solution. The temperature was controlled not to exceed 10°C during quenching, and then filtered through diatomaceous earth. After filtration, the crude product was concentrated under reduced pressure to obtain the crude product. The crude product was separated by silica gel column (SiO2, PE:EA=83:17). The reaction obtained a colorless oily product (3-bromo-4-(trifluoromethyl)phenyl)methanol (13.89 g, 77%).

[0407] Step B: At 0°C, N-bromosuccinimide (19.39 g, 108.9 mmol) was added to dichloromethane (200 mL) in which (3-bromo-4-(trifluoromethyl)phenyl)methanol (13.89 g, 54.46 mmol) and triphenylphosphine (28.57 g, 108.9 mmol) were dissolved. After the addition was complete, the entire mixture was stirred at room temperature for 2 hours. The reaction solution was filtered through diatomaceous earth and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column (SiO2, PE:EA=95:5). The reaction obtained a colorless oily product 2-bromo-4-(bromomethyl)-1-(trifluoromethyl)benzene (13.5 g, 78%).

[0408] Step C: At room temperature, cesium carbonate (20.75 g, 63.69 mmol) was added to N,N-dimethylformamide (300 mL) in which 2-bromo-4-(bromomethyl)-1-(trifluoromethyl)benzene (13.5 g, 42.46 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (7.20 g, 42.46 mmol) were dissolved. After the addition, the whole mixture was stirred at room temperature overnight. The reaction solution was filtered through diatomaceous earth and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column (SiO2, DCM:MeOH=95:5). The reaction obtained a brown oily product 1-(3-bromo-4-(trifluoromethyl)benzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (11.35 g, 66%). ESI-MS m / z=407.8[M+H] + ;Calcu.=407.0.

[0409] Step D: At room temperature and under nitrogen protection, 1-(3-bromo-4-(trifluoromethyl)benzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (11.35 g, 27.92 mmol), potassium carbonate (11.57 g, 83.74 mmol), (E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (1.07 g, 5.411 mmol) were dissolved in H2O (100 mL) and dioxane (400 mL). After the addition was complete, dichloro(1,1'-bis(diphenylphosphino)ferrocene)palladium (1.02 g, 1.396 mmol) was added, and the whole mixture was heated to 80°C and stirred overnight under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column (SiO2, PE:EA=50:50). The reaction gave a yellow solid product (E)-6-chloro-1-(3-(2-ethoxyvinyl)-4-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (6.6 g, 59%). ESI-MS m / z=397.9[M+H] + ;Calcu.=397.1.

[0410] Step E: At 0°C, add 48% hydrobromic acid (132 mL) to water (132 mL) and acetone (132 mL) in which (E)-6-chloro-1-(3-(2-ethoxyvinyl)-4-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (6.6 g, 16.59 mmol) is dissolved. After the addition is complete, the entire mixture is stirred at room temperature overnight. The reaction solution is temperature controlled at 0°C, and then the quenching reagent sodium hydroxide solution is slowly added until it is neutral. The temperature is controlled not to exceed 10°C during quenching. Extract with ethyl acetate three times (50 mL*3), combine the organic phases, wash once with saturated brine (50 mL), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain a crude product. The reaction afforded a colorless oily crude product of 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenyl)acetaldehyde (5.4 g, crude product). ESI-MS m / z=369.9 [M+H] + ;Calcu.=369.1.

[0411] Step F: At 0°C, sodium borohydride (1.38 g, 36.51 mmol) was added to tetrahydrofuran (300 mL) in which crude 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenyl)acetaldehyde (5.4 g, 14.60 mmol) was dissolved. After the addition was complete, the entire mixture was stirred at room temperature for 2 hours. The reaction solution was temperature controlled at 0°C, and then the quenching reagent methanol was slowly added until no obvious bubbles appeared in the reaction solution. The temperature was controlled not to exceed 10°C during quenching, and then filtered through diatomaceous earth. After filtration, the crude product was concentrated under reduced pressure to obtain the crude product. The crude product was separated by silica gel column (SiO2, DCM:MeOH=90:10). The reaction gave a white solid product, 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenyl)ethan-1-ol (1.7 g, two-step yield 27%). ESI-MS m / z=371.9 [M+H] + ;Calcu.=371.1.

[0412] Step G: At 0°C, p-toluenesulfonyl chloride (1.29 g, 6.78 mmol) was added to a solution of 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenyl)ethane-1-ol (630 mg, 1.695 mmol), triethylamine (1.41 mL, 10.17 mmol), and 4-dimethylaminopyridine (20.71 mg, 0.170 mmol) in dichloromethane (20 mL). After the addition was complete, the whole mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure to obtain a brown crude product, which was separated by silica gel column (SiO2, DCM:MeOH=95:5). The reaction gave a light yellow solid product 5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl-4-methylbenzenesulfonate (700 mg, 79%). ESI-MS m / z=525.9[M+H] + ;Calcu.=525.1.

[0413] Step H: To N,N-dimethylformamide (40 mL) in which 5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl-4-methylbenzenesulfonate (400 mg, 0.761 mmol) and methyl 6-oxo-1,6-dihydropyridazine-3-carboxylate (117 mg, 0.761 mmol) were dissolved, cesium carbonate (372 mg, 1.141 mmol) was added at room temperature, and the whole mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a yellow crude product, and water was added to precipitate a solid, which was filtered and freeze-dried to obtain a crude product of 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid methyl ester (309 mg, crude product). ESI-MS m / z=507.9[M+H] + ;Calcu.=507.1.

[0414] Step I: At 0°C, sodium borohydride (230 mg, 6.08 mmol) was added to tetrahydrofuran (2 mL) and ethanol (18 mL) in which the crude product of the previous step, 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid methyl ester (309 mg, 0.608 mmol) and calcium chloride (135 mg, 1.216 mmol) were dissolved. After the addition was complete, the entire mixture was stirred at room temperature for 3 hours. The reaction solution was temperature controlled at 0°C, and then the quenching reagent methanol was slowly added until no obvious bubbles appeared in the reaction solution. The temperature was controlled not to exceed 10°C during quenching, and then filtered through diatomaceous earth, and concentrated under reduced pressure after filtration to obtain a crude product. Water was added to precipitate the solid, which was filtered and freeze-dried to obtain the crude product 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl)-6-(hydroxymethyl)pyridazin-3(2H)-one (294 mg, crude product). ESI-MS m / z=479.9[M+H] + ;Calcu.=479.1.

[0415] Step J: At room temperature, cesium carbonate (2.0 g, 6.13 mmol) was added to acetonitrile (30 mL) in which 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl)-6-(hydroxymethyl)pyridazin-3(2H)-one (294 mg, 0.613 mmol) and lithium iodide (820 mg, 6.13 mmol) were dissolved. After the addition was complete, the whole mixture was heated to 110°C and stirred for 8 hours. It was then concentrated under reduced pressure to obtain a yellow crude product. The crude product was subjected to HPLC preparative separation, preparation conditions: Waters XBridge C18 (10 mm×150 mm, 5 μm); A: 5 mM HCOOH in water, B: MeCN; 5 mL / min; 0-3 min, 5% B; 3-15 min, 5%-50% B; detector, UV 254 nm). The reaction yielded compound C-37, a white solid product (E)-1 4 -Amino-3 4 -(Trifluoromethyl)-6 1 ,6 6 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3-benzocyclooctane-6 6 -ketone (18.60 mg, three-step yield 6%). 1H NMR(DMSO-d6,500MHz)δppm 7.95(s,1H),7.92(s,1H),7.84(br s,1H),7.61(d,J=8.1Hz,2H),7.39(d,J=8.2Hz,1H),7.36(d,J=9.6Hz,1H),6.83( d,J=9.6Hz,1H),5.37(s,2H),5.32(s,2H),4.56-4.62(m,2H),3.46-3.50(m,2H). ESI-MS m / z=444.0[M+H] + ;Calcu.=443.1.

[0416] Example 31: Synthesis of Compound C-38

[0417]

[0418] Step A: At room temperature, 5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl-4-methylbenzenesulfonate (300 mg, 0.57 mmol) and 6-oxo-1,6-dihydropyridine-3-carbaldehyde (70.2 mg, 0.570 mmol) were dissolved in N,N-dimethylformamide (30 mL), and cesium carbonate (279 mg, 0.856 mmol) was added. The whole mixture was stirred at room temperature overnight. Then it was concentrated under reduced pressure to obtain a yellow crude product, 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl)-6-oxo-1,6-dihydropyridine-3-carbaldehyde (900 mg, crude). ESI-MS m / z=476.9[M+H] + ;Calcu.=476.1.

[0419] Step B: At 0°C, sodium borohydride (182 mg, 4.82 mmol) was added to tetrahydrofuran (30 mL) in which the crude product 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl)-6-oxo-1,6-dihydropyridine-3-carboxaldehyde from the previous step was dissolved. After the addition was complete, the entire mixture was stirred at room temperature for 3 hours. The reaction solution was temperature controlled at 0°C, and then methanol was slowly added until no obvious bubbles appeared in the reaction solution. The temperature was controlled not to exceed 10°C during quenching, and then filtered through diatomaceous earth, and concentrated under reduced pressure after filtration to obtain a solid residue. The residue was dissolved in N,N-dimethylformamide and subjected to reverse phase separation. Chromatographic column: X select C18 (19mm x 150mm); mobile phase: water (containing 0.1% HCOOH) and acetonitrile; flow rate: 100 ml / min; gradient: acetonitrile increased from 0% to 50% in 30 minutes; detection wavelength: 254 nm. Freeze-dried under reduced pressure to obtain a light yellow product 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one (93 mg, 0.194 mmol, two-step yield 34%). ESI-MS m / z=479.0[M+H]+; Calcu.=478.1.

[0420] Step C: At room temperature, cesium carbonate (633 mg, 1.94 mmol) was added to acetonitrile (20 mL) in which 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one (93 mg, 0.194 mmol) and lithium iodide (260 mg, 1.94 mmol) were dissolved. After the addition was complete, the entire mixture was heated to or cooled to 110° C. and stirred for 8 hours. It was then filtered and concentrated under reduced pressure to obtain a yellow crude product. The crude product was subjected to HPLC preparation and separation, preparation conditions: Waters XBridge C18 (10mm×150mm, 5μm); A: 5mM NH3in water, B: MeCN; 5mL / min; 0~3min, 5% B; 3~15min, 5%~50% B; detector, UV 254nm). The reaction obtained compound C-38, white solid product (E)-1 4 -Amino-3 4 -(Trifluoromethyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-62 -Ketone (17.80 mg, 21%). 1 H NMR(DMSO-d6,500MHz)δppm8.30(d,J=2.3Hz,1H),8.04(s,1H),7.99(s,1H),7.75(br s,1H),7.51(d,J=8.2Hz,2H),7.36(dd,J=2.3,9.3Hz,1H),7.08(d,J=8.1Hz,1H),6.27 (d, J = 9.3Hz, 1H), 5.56 (s, 2H), 5.04 (s, 2H), 4.44 (t, J = 5.4Hz, 2H), 3.40-3.45 (m, 2H). ESI-MS m / z=443.2[M+H] + ;Calcu.=442.1.

[0421] Example 32: Synthesis of Compound C-39

[0422]

[0423] Step A: At room temperature, add (E)-1 to the Schlenk reaction tube in sequence. 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)benzocyclooctane-6 2-ketone (19 mg, 0.042 mmol), cyclopropylboronic acid (24 mg, 0.279 mmol), tris(dibenzylideneacetone)dipalladium (14 mg, 0.015 mmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (30 mg, 0.063 mmol), potassium carbonate (34 mg, 0.246 mmol). Then, 1,4-dioxane (4.0 mL) and water (1.0 mL) were added. After stirring evenly, degassing (degas) was performed three times to remove oxygen from the solvent. Then, under nitrogen protection, the reaction mixture was heated and stirred at 90 ° C for 24 hours. Afterwards, it was filtered through a sand core funnel with a small amount of silica gel, the solid was washed with methanol (20 mL), the obtained liquid was concentrated under reduced pressure to obtain a crude product, and the obtained solid was separated and purified by preparative chromatography. The chromatographic conditions are as follows: Welch C18 OBD (30 mm × 250 mm, 5 μm), A: 0.05% Am, B: ACN; 40 mL / min; 0-3.0 min, 5% B; 3.0-3.5 min, 5%-15% B; 3.5-15 min, 15%-75%; 15-17 min, 75%-100% B; detector, UV 254 nm). After freeze-drying, compound C-39 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -Cyclopropyl-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (0.26 mg, 1%). 1 H NMR (500MHz, DMSO-d6) δppm 8.29 (d, J=2.5Hz, 1H), 7.92 (s, 1H), 7.62-7.70 (m, 2H), 7.44 (br s,1H),7.35(dd,J=9.0,2.5Hz,1H),6.91-6.93(m,1H),6.82(d,J=7.5Hz,1H),6.26(d,J=9.0Hz,1H),5.38(s,2H ),5.08(s,2H),4.39-4.43(m,2H),3.37-3.40(m,2H),1.73-1.79(m,1H),0.80-0.85(m,2H),0.40-0.47(m,2H). ESI-MS m / z=415.3[M+H] + ;Calcu.=414.2.

[0424] Example 33: Synthesis of Compound C-40

[0425]

[0426] Step A: At room temperature and under nitrogen protection, dissolve (E)-1 4 -Amino-3 4 -Br-6 1 ,6 2 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -ketone (100mg, 0.221mmol) in dimethyl sulfoxide (5mL), add cuprous iodide (84mg, 0.442mmol) and triethylenediamine (99mg, 0.884mmol). After the addition, the whole mixture was heated to 145°C and stirred for 18 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a brown liquid crude residue, and the residue was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (19mm×250mm, 5μm); A: 0.05% ammonia in water, B: MeCN; 18mL / min; 0~3min, 5% B; 3~3.5min, 5%~15% B; 3.5~13min, 15%~15% B; 13~13.5min, 15%~100% B; detector, UV 254nm. Compound C-40 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -(Methylsulfinyl)-6 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 -Ketone (1.62 mg, 2%). 1 H NMR (500MHz, DMSO-d6) δppm 8.18(s,1H),7.80(s,1H),7.69(d,J=9.5Hz,1H),7.23(d,J=8.2Hz,1H),7.20(br s,1H),7.10(d,J=8.2Hz,1H),7.02(s,1H),6.66(brs,1H),6.44(d,J=9.5Hz,1H),5.30-5 .35(m,1H),5.05-5.15(m,3H),4.16(t,J=7.3Hz,2H),3.03(t,J=7.3Hz,2H),2.64(s,3H). ESI-MS m / z=437.2[M+H] +;Calcu.=436.1.

[0427] Example 34: Synthesis of Compound C-41

[0428]

[0429] Step A: Add imidazole (16.45 g, 241.7 mmol) and tert-butyldimethylsilyl chloride (20.03 g, 132.9 mmol, 16.4 mL) to a solution of (5-amino-3-pyridyl)methanol (15 g, 120.8 mmol) in dichloromethane (300 mL). Stir at 25°C for 12 hours. Dilute with water (200 mL), extract with dichloromethane (200 mL*3), dry the organic layer over magnesium sulfate, filter and concentrate. Compound 5-((tert-butyl(dimethyl)silyl)oxymethyl)pyridin-3-amine (27.7 g, 96%) is obtained as a white solid. 1 H NMR: (400MHz, DMSO-d6) δ7.82 (d, J = 2.3 Hz, 1H), 7.67 (s, 1H), 6.85 (s, 1H), 5.35 (brs, 2H), 4.59 (s, 2H), 0.89 (s, 9H), 0.10-0.01 (m, 6H).

[0430] Step B: Add di-tert-butyl dicarbonate (25.36 g, 116.2 mmol, 26.7 mL) to an ethanol solution (500 mL) of 5-((tert-butyl(dimethyl)silyl)oxymethyl)pyridin-3-amine (27.7 g, 116.2 mmol). Stir at 25°C for 12 hours. Dilute with water (150 mL), extract with ethyl acetate (300 mL*3), dry the organic layer over magnesium sulfate, filter and concentrate. Use flash silica gel chromatography ( 330g Purification by silica gel column, eluent: 0-15% ethyl acetate / petroleum ether gradient @100 mL / min) gave tert-butyl N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)carbamate (14.3 g, 36%) as a white solid compound. 1 H NMR (400MHz, DMSO-d6) δ9.59(s,1H),8.51(d,J=2.3Hz,1H),8.11(d,J=1.8Hz,1H),7.85(br s,1H),4.70(s,2H),1.48(s,9H),0.90(s,9H),0.08(s,6H).

[0431] Step C: Sodium borohydride (3.11 g, 82.29 mmol, 2 eq) was added in batches to a methanol solution (100 mL) of methyl 4-bromo-3-formylbenzoate (10 g, 41.14 mmol). The mixture was stirred at 25°C for 2 hours. The mixture was diluted with water (50 mL) at 0°C, extracted with ethyl acetate (50 mL*3), and dried over magnesium sulfate. The mixture was filtered and concentrated under reduced pressure to obtain the compound methyl 4-bromo-3-(hydroxymethyl)benzoate (9.76 g, 39.83 mmol, yield 97%) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.12 (d, J = 1.1 Hz, 1H), 7.80-7.60 (m, 2H), 5.63 (t, J = 5.6 Hz, 1H), 4.54 (d, J = 5.6 Hz, 2H), 3.86 (s, 3H).

[0432] Step D: At 0°C, methanesulfonyl chloride (11.47 g, 100.1 mmol, 7.75 mL) was added to a solution of methyl 4-bromo-3-(hydroxymethyl)benzoate (9.76 g, 39.83 mmol) and triethylamine (11.47 g, 113.4 mmol, 15.8 mL) in dichloromethane (100 mL). The mixture was stirred at 0°C for 2 hours. The reaction mixture was quenched with water (100 mL) at 0°C and extracted with dichloromethane (100 mL*3). Drying over magnesium sulfate, filtering and concentrating under reduced pressure gave a white solid compound methyl 4-bromo-3-(methylsulfonyloxymethyl)benzoate (11.25 g, crude product).

[0433] Step E: Add tert-butyl N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)carbamate (4.82 g, 14.23 mmol) to N,N-dimethylformamide (100 mL), then add sodium hydride (1.14 g, 28.47 mmol, 60% purity) in batches, stir at 0°C for 30 minutes, then add methyl 4-bromo-3-(methylsulfonyloxymethyl)benzoate (4.6 g, 14.23 mmol). The mixture was stirred at 25°C for 2 hours. Quench with ice water (100 mL), extract with ethyl acetate (100 mL*3), dry the organic layer over magnesium sulfate, filter and concentrate. Use flash silica gel chromatography ( 80g The mixture was purified by silica gel column with 0-15% ethyl acetate / petroleum ether gradient @100 mL / min as the eluent to give methyl 4-bromo-3-((tert-butyloxycarbonyl-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridinyl)amino)methyl)benzoate (4.99 g, 52% yield for two steps) as a white solid compound. 1H NMR (400MHz, DMSO-d6) δ8.44(d,J=2.4Hz,1H),8.28(d,J=1.5Hz,1H),7.89(s,1H),7.73-7.82(m, 2H),7.48(s,1H),4.95(s,2H),4.70(s,2H),3.83(s,3H),1.39(s,9H),0.81(s,9H),0.00(s,6H). ESI-MS m / z=566.8[M+H] + ;Calcu.=566.2.

[0434] Step F: Diisobutylaluminum hydride (1M, 38mL) was added dropwise to a tetrahydrofuran solution (40mL) of methyl 4-bromo-3-((tert-butyloxycarbonyl-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)amino)methyl)benzoate (4.3g, 7.60mmol) at 0°C, and the mixture was stirred at 25°C for 2 hours. Diluted with water (50mL), 1N dilute hydrochloric acid was added to pH=7, and then extracted with ethyl acetate (50mL*3), the organic layer was dried over magnesium sulfate, filtered and concentrated to obtain a white solid compound N-((2-bromo-5-(hydroxymethyl)phenyl)methyl)-N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)carbamic acid tert-butyl ester (3.2g, 78%). ESI-MS m / z=539.0[M+H] + ;Calcu.=538.2.

[0435] Step G: To a solution of tert-butyl N-((2-bromo-5-(hydroxymethyl)phenyl)methyl)-N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)carbamate (3.1 g, 5.77 mmol) in dichloromethane (20 mL), triphenylphosphine (1.74 g, 6.63 mmol) and carbon tetrabromide (2.29 g, 6.92 mmol) were added and stirred at 25°C for 12 hours. The resulting reaction solution, tert-butyl N-((2-bromo-5-(bromomethyl)phenyl)methyl)-N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)carbamate, was directly used in the next step without treatment and purification. ESI-MS m / z=601.1[M+H] + ;Calcu.=600.1.

[0436] Step H: The above reaction solution of tert-butyl N-((2-bromo-5-(bromomethyl)phenyl)methyl)-N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridinyl)carbamate (3.02 g, 5.03 mmol) was poured into N,N-dimethylformamide (20 mL), and then potassium carbonate (2.09 g, 15.09 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (810.22 mg, 4.78 mmol) were added and stirred at 25 °C for 12 hours. The reaction mixture was concentrated to give yellow oily substance tert-butyl-N-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenyl)methyl)-N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)carbamate (2.43 g, crude product). ESI-MS m / z=689.8[M+H] + ;Calcu.=689.1.

[0437] Step I: To a solution of tert-butyl N-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenyl)methyl)-N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridinyl)carbamate (2.43 g, 3.53 mmol) in tetrahydrofuran (30 mL), tetrabutylammonium fluoride (1 M, 5.29 mL) was added. Stir at 25°C for 2 hours. The mixture was concentrated at 50°C to give a white solid compound, tert-butyl N-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenyl)methyl)-N-(5-(hydroxymethyl)-3-pyridinyl)carbamate (1.1 g, 54%). ESI-MS m / z=575.9[M+H] + ;Calcu.=575.1.

[0438] Step J: To a solution of tert-butyl N-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenyl)methyl)-N-(5-(hydroxymethyl)-3-pyridinyl)carbamate (300 mg, 521.87 μmol) in N-methylpyrrolidone (20 mL), sodium hydride (62.62 mg, 1.57 mmol, 60% purity) was added and stirred at 110 ° C for 12 hours. The reaction mixture was quenched with water (5 mL) and extracted with ethyl acetate (5 mL*2). The combined organic layer was washed with brine (5 mL*2), dried over magnesium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The crude product was purified by thin layer chromatography to give compound M-3, 1 4 -Amino-3 4 -Bromo-1 1H-8-Oxa-5-aza-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(3,5)-pyridine-3(1,3)-benzoic acid cyclooctane-5-carboxylic acid tert-butyl ester (400 mg), yellow oil. ESI-MS m / z=539.8[M+H] + ;Calcu.=539.1.

[0439] Step K: 1 4 -Amino-3 4 -Bromo-1 1 H-8-oxa-5-aza-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(3,5)-pyridine-3(1,3)-benzoic acid cyclooctane-5-carboxylic acid tert-butyl ester (60 mg, 111.0 μmol), potassium trifluoro(pyrrolidin-1-ylmethyl)borate (63.63 mg, 333.1 μmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (5.3 mg, 11.1 μmol), tris(dibenzylideneacetone)dipalladium (10.2 mg, 11.1 μmol) and sodium carbonate (35.3 mg, 333.1 μmol) were added to a solution of tetrahydrofuran / water = 2:1 (2 ml), replaced with nitrogen three times, and then stirred at 80°C for 12 hours under nitrogen atmosphere. The reaction mixture was quenched with water (2 mL) and extracted with ethyl acetate (5 mL*2). The combined organic layers were washed with brine (5 mL*2), dried over magnesium sulfate, filtered and concentrated under reduced pressure to give compound M-4, tert-butyl 1 4 -Amino-3 4 -(pyrrolidin-1-ylmethyl)-1 1 H-8-Oxa-5-aza-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(3,5)-pyridine-3(1,3)-benzocyclooctanecarboxylic acid tert-butyl ester (60 mg, crude product). ESI-MS m / z=543.1[M+H] + ;Calcu.=542.3.

[0440] Step L: Go to 1 4 -Amino-3 4 -(pyrrolidin-1-ylmethyl)-1 1To tert-butyl H-8-oxa-5-aza-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(3,5)-pyridine-3(1,3)-benzocyclooctanecarboxylate (30 mg, 55.29 μmol) in dichloromethane (2 mL), N,N-diisopropylethylamine (35.7 mg, 276.4 μmol, 48 μL) and trimethylsilyl trifluoromethanesulfonate (61.4 mg, 276.4 μmol) were added, and the mixture was stirred at 25°C for 2 hours. The mixture was concentrated in a vacuum at 50°C and purified by reverse phase high performance liquid chromatography (chromatographic column: maximum chromatographic column C18 150*40mm*10um, mobile phase: [water (ammonium bicarbonate)-acetonitrile], gradient: 14%-54% B, 32 min) to obtain compound C-41 as a white solid 3 4 -(pyrrolidin-1-ylmethyl)-1 1 H-8-Oxa-5-aza-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(3,5)-pyridine-3(1,3)-benzocyclooctane-1 4 -amine (2.14 mg, 8%). 1 H NMR (400MHz, DMSO-d6) δ = 7.91 (s, 1H), 7.79 (s, 1H), 7.75-7.58 (m, 3H), 7.53 (br s,1H),7.18(d,J=7.6Hz,1H),7.12-7.07(m,1H),6.57-6.47(m,1H),6.13-6. 01(m,1H),5.39(s,2H),5.33(s,2H),4.43-4.33(m,2H),3.67(s,2H),2.40(br s,4H),1.73(brs,4H). ESI-MS m / z=443.2[M+H] + ;Calcu.=442.2.

[0441] Test Example 1: Evaluation of drug efficacy at the cellular level

[0442] Purpose: Using HEK-Blue TM hTLR7 in vitro cell model to evaluate the TLR7 and TLR8 agonist activity of compounds

[0443] Experimental Materials:

[0444] 1 Cell line: HEK-Blue TM hTLR7(InvivoGen), HEK-Blue TM hTLR8 (InvivoGen)

[0445] 2 Reagent: QUANTI-Blue TMSolution (InvivoGen), DMEM (Invitrogen), FBS (HyClone), L-glutamine (GIBCO), TrypLE Express Enzyme (GIBCO), DPBS (GIBCO), Normocin TM (InvivoGen);Penicillin-Streptomycin(GIBCO)

[0446] 3 Instruments: Microplate reader (PerkinElmer, Multimode plate reader 2014), CO2 incubator (THERMO, STERI-CYCLEI160), cell counter (BECKMAN COULTER, vi-cell XR)

[0447] Experimental methods:

[0448] Step 1: Cell plating

[0449] a) Discard the culture medium in the T150 flask, add 10 mL of pre-warmed DPBS, and gently shake the flask to rinse the cells. After discarding the DPBS, add 8.0 mL of TrypLE TM Add the Express enzyme to the T150 culture flask and gently shake the flask to allow it to infiltrate all cells.

[0450] b) Add TrypLE TM Express enzyme was discarded and the tube was placed in an incubator for 1 to 2 minutes.

[0451] c) Add serum-containing culture medium to terminate digestion, collect the cell suspension, centrifuge at 300r at room temperature for 5 minutes, and discard the supernatant.

[0452] d) Add fresh culture medium, mix the cells by pipetting, and then count them using a vi-cell XR cell counter.

[0453] e) Dilute the cells to a density of 6.25*10 5 cells / mL, 80 μL of the diluted cell suspension was inoculated into a 96-well plate, i.e., 5*10 4 cells / well.

[0454] f) Place the 96-well plate in an incubator and incubate overnight (5% CO2, 37°C).

[0455] Step 2: Drug treatment

[0456] a) The test compound was prepared into a 10 mM stock solution in DMSO;

[0457] b) The compound to be tested was diluted from 10 mM to 0.1 mM using culture medium, and then diluted 3-fold using culture medium containing 1% DMSO, for a total of 8 concentration gradients, with DMSO as negative control, and R848 and GS9620 as positive controls.

[0458] c) Take 20 μL of the diluted compound and add it to a 96-well plate inoculated with 80 μL of cells, dilute the drug 5 times, and the final concentration of DMSO in each well is 0.2%. The final concentrations of the test compound and the control compound are 20000, 6667, 2222, 741, 247, 82, 27, and 9 nM.

[0459] d) The 96-well plate was placed in a 5% CO2, 37°C incubator and incubated for 24 hours.

[0460] Step 3: Activity detection

[0461] a) Take 10 μL of the cell supernatant after compound incubation and transfer it to a 96-well transparent test plate, then add 90 μL of QUANTI-Blue to each well of the test plate. TM Solution, incubate in a 5% CO2, 37°C incubator for 1 hour.

[0462] b) The light absorption of secretory alkaline phosphatase (SEAP) at 650 nm was detected using an ELISA reader.

[0463] Data Analysis:

[0464] The activation factor was calculated using the following formula: activation factor = OD 650 Treatment group / OD 650 The GraphPadPrism 7 software was used to fit the curve according to the concentration of the compound and the corresponding activation factor, and the EC 50 value.

[0465] Reference substances: R848 (InvivoGen) and GS-9620 (CAYMAN), the structures are shown below:

[0466]

[0467] The results of hTLR7 activation experiment are shown in the following table:

[0468]

[0469]

[0470] The results of the compound activation experiment on hTLR8 are as follows:

[0471] serial number <![CDATA[hTLR8 EC 50 (nM) <!-- 60 -->]]> R848 4549 C-6 >20000 C-27 >20000 C-37 >20000 C-39 >20000

[0472] Test Example 2: Test on the agonistic activity of the compounds of the present invention on human peripheral blood mononuclear cells

[0473] Human peripheral blood mononuclear cells (hPBMCs) represent the main human immune cells in the blood, including lymphocytes, monocytes and dendritic cells. These cells express TLR7, TLR8 or TLR9 and respond to stimulation by the corresponding ligands. In this experiment, hPBMCs were stimulated with potential TLR7 agonists, and the levels of IFN-α and TNF-α were detected to reflect the activation of the compounds on the TLR7 receptor.

[0474] Experimental Materials:

[0475] 1. Cell line: hPBMCs (sailybio), culture conditions: RPMI 1640 medium + 10% (v / v) FBS + 1% double antibody (Penicillin-Streptomycin), 5% CO2, 37°C.

[0476] 2. Reagents: RPMI 1640 medium (GIBCO), FBS (GIBCO), Penicillin-Streptomycin (GIBCO), Human IFN-alpha 2 / IFNA2 DuoSet ELISA (R&D), Human TNF-alpha ELISA (R&D).

[0477] 3. Instruments: microplate reader (perkinElmer, Multimode plate reader 2014), CO2 incubator (THERMO, STERI-CYCLEI160), cell counter (BECKMAN COULTER, vi-cell XR).

[0478] Experimental methods:

[0479] a) The frozen hPBMCs were thawed in a 37°C water bath and transferred to 1640 complete medium. After centrifugation at 500r for 10 minutes at room temperature, the supernatant was discarded, the cells were resuspended in 1640 complete medium and transferred to a culture flask, and cultured overnight at 37°C and 5% CO2.

[0480] b) Collect hPBMCs after stable culture, count and dilute the cells to 1.11*10 7 cells / mL, 180 μL of the diluted cell suspension was inoculated into a 96-well plate, i.e. 2*10 6 cells / well, and the 96-well plate was placed in a constant temperature incubator and incubated for 24 h (5% CO2, 37°C).

[0481] c) The test compound was prepared into a 10 mM stock solution with DMSO and diluted to 0.01 mM (IFN-α detection) and 0.1 mM (TNF-α), respectively. The compound was then diluted into 8 concentration gradients with culture medium containing 2% DMSO.

[0482] d) 20 μL of the diluted compound was added to a 96-well cell culture plate, and the final concentration of DMSO in each well was 0.2%.

[0483] e) After incubation at 37° C. for 24 hours, the cell culture supernatant was collected by centrifugation, and the levels of IFN-α and TNF-α in the supernatant were determined using an ELISA detection kit.

[0484] f) The light absorption at 450 nm was measured by an enzyme-labeled instrument, and the level of secretion of the corresponding cytokines was quantitatively determined according to the corresponding standard curve, and the EC was calculated by fitting the curve. 50 value.

[0485] g) The experimental results are shown in the following table:

[0486]

Claims

1. A compound having a structure as shown in formula (I), a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or a metabolite thereof, in, R 1 Selected from: hydrogen, halogen, hydroxyl, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl; Q is selected from -O-, -S-, -NH-; Ring A is selected from phenyl, naphthyl, 5-6 membered heteroaryl and bicyclic heteroaryl, wherein the phenyl, naphthyl, 5-6 membered heteroaryl and bicyclic heteroaryl are optionally substituted by 1, 2 or 3 groups selected from the following: hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, =O, -NR 3 R 4 、-CH2R a 、-C(=O)R b 、-OR c , C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -(C1-C6 alkyl)OH, -(C1-C6 alkyl)NH2, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -S(=O)(=O)NH(C1-C3 alkyl), -NR d S(=O)(=O)(C1-C6 alkyl), -P(=O)R e R f , sulfonic acid, -N=NPh, -Si(C1-C3 alkyl)3, -C(O-C1-C6 alkyl)=CH2; Among them, R a , R b , R c , R d , R e , R f Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C(=O)(C1-C6 alkyl); wherein R 3 , R 4 are each independently selected from hydrogen, C 1-6 alkyl, C(═O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy; Ring B is selected from 5-6 membered nitrogen-containing heteroaryl and bicyclic heteroaryl, and the 5-6 membered nitrogen-containing heteroaryl and bicyclic heteroaryl are optionally substituted by 1, 2 or 3 groups selected from the following: hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)R b’ 、-OR c’ , C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -(C1-C6 alkyl)OH, -(C1-C6 alkyl)NH2, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -S(=O)(=O)NH(C1-C3 alkyl), -NR d’ S(=O)(=O)(C1-C6 alkyl), -P(=O)R e’ R f’ , sulfonic acid, -N=NPh, -Si(C1-C3 alkyl)3; wherein R a’ , R b’ , R c’ , R d’ , R e’ , R f’ Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C(=O)(C1-C6 alkyl); wherein R 3’ , R 4’ Each is independently selected from hydrogen, C1-C6 alkyl, C(═O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy; L is selected from -(CH2) m -W-(CH2) n -, wherein W is selected from -O-, -NH-, -N(Boc)-, -NHC(=O)-, -CH2-, -CH(C1-C6 alkyl)-, -C(C1-C6 alkyl)(C1-C6 alkyl)-, -C(=O)-, -C(=O)NH-, -N(C1-C6 alkyl)-, -N(C1-C6 alkyl)C(=O)-, -C(=O)N(C1-C6 alkyl)-, -S-, -S(=O)-, -S(=O)(=O)-, -P -(=O)(C1-C6 alkyl)-, -P(=O)(C1-C6 alkoxy)-, -P(=O)(OH)-, -OC(=O)-, -C(=O)O-, -NHS(=O)(=O)-, -S(=O)(=O)NH-, -N(C1-C6 alkyl)S(=O)(=O)-, -S(=O)(=O)N(C1-C6 alkyl)-, m and n are each independently selected from an integer between 0 and 6 (e.g., 0, 1, 2, 3, 4, 5 or 6); T is selected from -(CH-T 1 ) p -(Z) q -(CH-T 2 ) r -, wherein Z is selected from -O-, -NH-, -NHC(=O)-, -CH2-, -CH(C1-C6 alkyl)-, -C(C1-C6 alkyl)(C1-C6 alkyl)-, -C(=O)-, -C(=O)NH-, -N(C1-C6 alkyl)-, -N(C1-C6 alkyl)C(=O)-, -C(=O)N(C1-C6 alkyl)-, -S-, -S(=O)-, -S(=O)(=O)-, -P(=O)(C1-C6 alkyl)- wherein p, q, and r are each independently selected from an integer between 0 and 6 (e.g., 0, 1, 2, 3, 4, 5, or 6), and p and r cannot be 0 at the same time, and T 1 , T 2 Each is independently selected from: hydrogen, halogen, nitro, hydroxyl, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, mercapto, -S-(C1-C6 alkyl).

2. The compound of claim 1, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein: R 1 Selected from: hydrogen, halogen, C1-C6 alkyl, hydroxyl.

3. The compound of claim 1 or 2, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein: Q is -O-.

4. The compound of any one of claims 1 to 3, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein: Ring A is phenyl or 5-6 membered heteroaryl, and optionally, Ring A is substituted by 1, 2 or 3 substituents; Preferably, ring A is phenyl, and optionally, ring A is substituted by 1, 2 or 3 substituents; Preferably, the substituents of ring A are selected from the following groups: hydrogen, halogen, cyano, nitro, hydroxyl, C1-C6 alkyl, halogenated C1-C6 alkyl, =O, -NR 3 R 4 、-CH2R a 、-C(=O)R b 、-OR c , C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -S(=O)(=O)NH(C1-C3 alkyl), -NR d S(=O)(=O)(C1-C6 alkyl), -P(=O)R e R f , -C(O-C1-C6 alkyl)=CH2; Among them, R a , R b , R c , R d , R e , R f Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, -C(=O)(C1-C6 alkyl); wherein R 3 , R 4 are each independently selected from hydrogen, C 1-6 alkyl, -C(=O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy; Preferably, the substituents of ring A are selected from the following groups: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, halogenated C1-C6 alkyl, =O, -CH2R a 、-C(=O)R b 、-OR c , C3-C6 saturated cycloalkyl, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NR d S(=O)(=O)(C1-C6 alkyl), -P(=O)R e R f , -C(O-C1-C6 alkyl)=CH2; wherein R a , R b , R c , R d , R e , R f Each is independently selected from hydrogen, -OH, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -N(C1-C6 alkyl)2; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, hydroxyl, C1-C6 alkyl, halogenated C1-C6 alkyl; Preferably, R a , R b , R c , R d , R e , R f Each is independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -N(C1-C6 alkyl)2; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

5. The compound of any one of claims 1 to 4, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein: The ring B is a 5-6 membered monoheteroaryl group containing 1, 2 or 3 nitrogen atoms; Preferably, the 5-6 membered monoheteroaryl group containing 1, 2 or 3 nitrogen atoms is selected from the following heteroaromatic ring-forming groups: Preferably, ring B is Optionally, Ring B is substituted with 1, 2 or 3 substituents; Preferably, ring B is Optionally, Ring B is substituted with 1, 2 or 3 substituents; Preferably, the substituents of ring B are selected from the following groups: hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)R b’ 、-OR c’ , C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -(C1-C6 alkyl)OH, -(C1-C6 alkyl)NH2, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -S(=O)(=O)NH(C1-C3 alkyl), -NR d’ S(=O)(=O)(C1-C6 alkyl), -P(=O)R e’ R f’ , sulfonic acid, -N=NPh, -Si(C1-C3 alkyl)3; Among them, R a’ , R b’ , R c’ , R d’ , R e’ , R f’ Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C(=O)(C1-C6 alkyl); wherein R 3’ , R 4’ Each is independently selected from hydrogen, C1-C6 alkyl, C(═O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy; Preferably, the substituents of ring B are selected from the following groups: hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)R b’ 、-OR c’ , C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group; wherein R a’ , R b’ , R c’ Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C(=O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy; Preferably, the substituents of ring B are selected from the following groups: hydrogen, =0, C1-C6 alkyl.

6. The compound of any one of claims 1 to 5, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein: L is selected from -(CH2) m -W-(CH2) n -, wherein W is selected from -O-, -NH-, -N(Boc)-, -NHC(=O), -CH2-, -CH(C1-C6 alkyl)-, -C(C1-C6 alkyl)(C1-C6 alkyl)-, -C(=O)-, -C(=O)NH-, -N(C1-C6 alkyl)-, -S-, -S(=O)-, -S(=O)(=O)-, -P(=O)(OH)-, -OC(=O)-, -C(=O)O-, -NHS(=O)(=O)-, -S(=O)(=O)NH-, and m and n are each independently selected from 0, 1 or 2; Preferably, L is selected from the following structures: -O-CH2-, -NH-CH2-, -CH2-, -NH-, -NH-C(=O)-, -CH2-O-, -CH2-O-CH2-, -CH2-CH2-, -O-CH2-CH2-, -N(Boc)-CH2-; Preferably, L is selected from the following structures: -O-CH2-, -CH2-O-, -NH-CH2-, -CH2-NH-, -CH2-CH2-, -N(Boc)-CH2-; Preferably, L is selected from the following structures: -NH-CH2-, -CH2-CH2-, -N(Boc)-CH2-.

7. The compound of any one of claims 1 to 6, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein: q is 0.

8. The compound of any one of claims 1 to 7, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein: T is selected from -(CH-T 1 ) p -, where p is an integer between 1 and 6, T 1 Selected from: hydrogen, halogen, nitro, hydroxyl, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, mercapto, -S-(C1-C6 alkyl); Preferably, T 1 Selected from hydrogen, C1-C6 alkyl; Preferably, T is selected from -CH2-.

9. The compound according to any one of claims 1 to 8, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein the compound has a structure as shown in formula (I'), in, R 1 , Q, L, Ring A, and Ring B are as defined in any one of claims 1 to 8; R 2 Selected from hydrogen or C1-C6 alkyl.

10. The compound according to any one of claims 1 to 9, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein the compound has a structure as shown in formula (II): in, R 1 , Q, L are as defined in any one of claims 1 to 8 respectively; R 2 Selected from hydrogen or C1-C6 alkyl; R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

11. The compound according to any one of claims 1 to 10, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein the compound has a structure as shown in formula (II'): R 1 , R 2 , Q, L are as defined in any one of claims 1 to 8 respectively; R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein, R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

12. The compound according to any one of claims 1 to 11, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein the compound has a structure as shown in formula (II"): R 1 , R 2 , Q, L are as defined in any one of claims 1 to 8 respectively; R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein, R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

13. A compound according to any one of claims 1 to 9, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or a metabolite thereof, wherein the compound has a structure as shown in formula (III): in, R 1 , Q, L are as defined in any one of claims 1 to 8 respectively; R 2 Selected from hydrogen or C1-C6 alkyl; R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

14. The compound according to any one of claims 1 to 9 or 13, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein the compound has a structure as shown in formula (III'): R 1 , R 2 , Q, L are as defined in any one of claims 1 to 8 respectively; R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein, R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

15. A compound according to any one of claims 1 to 9, 13 or 14, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or a metabolite thereof, wherein the compound has a structure as shown in formula (III"): R 1 , R 2 , Q, L are as defined in any one of claims 1 to 8 respectively; R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein, R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl.

16. The compound according to any one of claims 1 to 15, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein the compound has a structure as shown in formula (V): in, R 1 , Q, L as defined in any one of claims 1-15; X is C or N; R 2 Selected from hydrogen or C1-C6 alkyl; R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-NR 3 R 4 , -C(=O)(C1-C6 alkyl), cyano, -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, 3-6 membered cycloalkyl, -S(C1-C3 alkyl), -P(=O)(C1-C3 alkyl)(C1-C3 alkyl); wherein R 3 , R 4 Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl; Preferably, R 5 , R 6 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered cycloalkyl, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-N(C1-C3 alkyl)(C1-C3 alkyl), cyano, -P(=O)(C1-C3 alkyl)(C1-C3 alkyl), -NHS(=O)(=O)(C1-C6 alkyl), -C(O-C1-C6 alkyl)=CH2, -C(=O)(C1-C6 alkyl), -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), halogenated C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, C1-C6 alkyl; Preferably, the -CH2-(3-6 membered nitrogen-containing heterocyclic group) is Preferably, R 7 is selected from hydrogen, C1-C6 alkyl (e.g. methyl); Preferably, R 7 For hydrogen.

17. The compound according to any one of claims 1 to 16, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, wherein the compound has a structure as shown in formula (V'): R 1 , R 2 , Q, L, X, R 5 , R 6 , R 7 As defined in any one of claims 1 to 16 respectively.

18. The compound according to any one of claims 1 to 17, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite thereof, which has a structure as shown in formula (V"): R 1 , R 2 , Q, L, X, R 5 , R 6 , R 7 As defined in any one of claims 1 to 16 respectively.

19. The compound according to any one of claims 1 to 18, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof, which has a structure as shown in formula (V''): R 1 , R 2 , Q, L, and X are respectively as defined in any one of claims 1 to 16; R 5 is hydrogen; R 6 Selected from: hydrogen, halogen, 3-6 membered cycloalkyl, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-N(C1-C3 alkyl)(C1-C3 alkyl), cyano, -P(=O)(C1-C3 alkyl)(C1-C3 alkyl), -C(O-C1-C6 alkyl)=CH2, -C(=O)(C1-C6 alkyl), -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), halogenated C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from: hydrogen, C1-C6 alkyl; Preferably, R 6 Selected from: -Br, cyclopropyl, -CH2-N(CH3)(CH3), cyano, -P(=O)(CH3)(CH3), -C(OEt)=CH2, -C(=O)CH3, -SCH3, -S(=O)CH3, -S(=O)(=O)CH3, -CF3; R 7 Selected from hydrogen or C1-C6 alkyl.

20. A compound according to any one of claims 1 to 19, a pharmaceutically acceptable salt, a prodrug, a stereoisomer, a tautomer, a hydrate, a solvate, a crystal form, an isotope-labeled substance or a metabolite thereof, wherein the hydroxyl group, -NH- or -NH2 in the compound is protected by a substituent (e.g., Boc, TBDPS).

21. A compound according to any one of claims 1 to 20, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystalline form, isotope-labeled substance or a metabolite thereof, wherein the compound has a structure selected from the following:

22. A pharmaceutical composition comprising a compound according to any one of claims 1 to 21, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or a metabolite thereof; optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

23. Use of the compound of any one of claims 1 to 21, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite thereof in the preparation of a medicament, wherein the medicament is used as a TLR7 agonist for preventing or treating a disease associated with TLR7; Preferably, the disease associated with TLR7 is selected from: (1) Tumors, including but not limited to: breast cancer, melanoma, meningioma, soft tissue sarcoma, salivary gland tumors, intraspinal tumors, mediastinal tumors, brain cancer, bone cancer, penile cancer, osteosarcoma, intracranial tumors, tongue cancer, maxillary sinus cancer, thyroid cancer, malignant lymphoma, multiple myeloma, pituitary adenoma, testicular tumors, non-Hodgkin's lymphoma, bladder cancer, leukemia, gastric cancer, nasopharyngeal cancer, laryngeal cancer, oral cancer, esophageal cancer, lung cancer, kidney cancer, cervical cancer, choriocarcinoma, vulvar cancer, skin cancer, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, rectal cancer, colorectal cancer, Kaposi's sarcoma, non-melanoma skin cancer (including squamous cell carcinoma and basal cell carcinoma), hemangioma, glioma, non-Hodgkin's lymphoma (diffuse large B-cell lymphoma, B-cell non-Hodgkin's lymphoma), colon cancer, liver cancer, neuroblastoma, glioblastoma; (2) diseases associated with viral infection, including but not limited to dengue virus, yellow fever virus, West Nile virus, Japanese encephalitis virus, tick-borne encephalitis virus, Kunjin virus, Murray Valley encephalitis virus, St. Louis encephalitis virus, Omsk hemorrhagic fever virus, bovine viral diarrhea virus, Zika virus, HIV, HBV, HCV, HPV, RSV, SARS or influenza virus; (3) Inflammatory diseases, respiratory diseases, skin diseases, gastrointestinal diseases, hepatobiliary diseases, nervous system diseases, musculoskeletal and connective tissue diseases or autoimmune diseases, such as asthma, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome (ARDS), ulcerative colitis, Crohn's disease, bronchitis, dermatitis, actinic keratosis, allergic rhinitis, psoriasis, scleroderma, urticaria, rheumatoid arthritis, multiple sclerosis, lupus, drug-induced allergic reaction, rheumatism.

24. Use of the compound according to any one of claims 1 to 21, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof as a vaccine adjuvant.

25. Use of the compound according to any one of claims 1 to 21, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof as a photodynamic therapy agent.

26. Use of the compound of any one of claims 1 to 21, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof as a conjugate drug (e.g., antibody immunostimulatory drug conjugate ISAC).

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