Macrocyclic pyrazolopyrimidine compound and use thereof

Abstract

The present application relates to the field of biomedicine, and in particular to a macrocyclic pyrazolopyrimidine compound which has better immunomodulatory activity and selectivity. The present invention further provides a use of the macrocyclic pyrazolopyrimidine compound for preventing or treating a TLR7-related disease, and a use as a vaccine adjuvant, a photodynamic therapeutic agent, and a drug conjugate.

Description

Macrocyclic pyrazolopyrimidine compounds and uses thereof

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application is based on the application with CN application number 202311138066.2 and application date September 4, 2023, and claims its priority. The content of the CN application is hereby introduced as a whole into this application. Technical Field

[0003] The present application relates to the field of biomedicine, and in particular to macrocyclic pyrazolopyrimidine compounds and uses thereof. Background Art

[0004] 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 primary immune strategies of vertebrates are innate immunity and adaptive immunity, and the innate immune system is key to microbial recognition and clearance, inflammatory responses, and cell death pathways. Targeting the proteins and signaling pathways of the innate immune system to develop therapeutics for diseases offers numerous possibilities for drug development.

[0005] Toll-like receptors (TLRs) are a family of proteins that play a crucial role in the immune system and are key receptors in the innate immune system. As pattern recognition receptors (PRRs), TLRs can recognize damage-associated molecular patterns (DAMPs) and pathogen-associated molecular patterns (PAMPs), inducing activation of the NF-κB pathway and the secretion of cytokines and chemokines, thereby activating both innate and adaptive immune responses. Currently, 13 members of the TLR family have been discovered (TLR1-13), of which 10 are human (TLR1-10), primarily distributed on immune cells. Different TLR subtypes have specific binding ligands. Because TLRs are key molecules in immune triggering, TLR targets hold significant value in anti-infection, anti-inflammatory, and anti-tumor efforts.

[0006] The TLR7 gene is located at Xp22.3 on chromosome X. The TLR7 protein, known as a nucleic acid sensor, is localized to the endosomal membrane and can recognize nucleotides and nucleosides associated with intracellular pathogens. It is activated in the presence of viral single-stranded RNA (ssRNA). The TLR7 molecule has two binding sites, each of which binds to a specific ligand. The first binding site can bind to single nucleosides such as adenosine and guanosine, as well as small nucleoside analogs (such as the TLR7 / 8 agonist R848). The second site binds to ssRNA and can influence the affinity between the first binding site and the ligand. Ligand binding to TLR7 activates the MyD88 pathway, inducing cytokine secretion through a MyD88-NF-κB-dependent signaling cascade, thereby activating the immune response.

[0007] 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 as vaccine adjuvants. Currently, multiple agonists targeting TLR7 have entered clinical research, but the only small molecule compound currently 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 effect or immune-related adverse reactions. Therefore, the design of new, efficient, and selective TLR7 small molecule agonists has important clinical value.

[0008] Summary of the Invention

[0009] The present invention aims to provide a novel macrocyclic pyrazole TLR7 small molecule agonist with enhanced 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 TLR7-related diseases, as well as its use as a vaccine adjuvant and photodynamic therapy agent. The following inventions are provided:

[0010] Compound

[0011] 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 metabolite thereof,

[0012] in,

[0013] R 1 Selected from: hydrogen, halogen, hydroxy, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl;

[0014] Q is selected from -O-, -S-, and -NH-;

[0015] 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, 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, -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 group consisting of hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, and C1-C6 alkoxy;

[0016] Ring B is selected from 5-6 membered nitrogen-containing heteroaryl and bicyclic heteroaryl, wherein 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, 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, 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 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, halo-substituted C1-C6 alkyl, C1-C6 alkoxy;

[0017] 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);

[0018] 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) -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)-, p, q, r are each independently selected from an integer between 0 and 6 (for example, 0, 1, 2, 3, 4, 5 or 6), and p and r cannot be 0 at the same time, T 1 、T 2 Each is independently selected from: hydrogen, halogen, nitro, hydroxy, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, mercapto, -S-(C1-C6 alkyl).

[0019] In certain embodiments, R 1 Selected from: hydrogen, halogen, C1-C6 alkyl, hydroxyl.

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

[0021] In certain embodiments, R 1 Selected from hydrogen and halogen.

[0022] In certain embodiments, R 1 Selected from hydrogen.

[0023] In certain embodiments, Q is -O-.

[0024] In certain embodiments, Ring A is a 5-6 membered heteroaryl, optionally substituted with 1, 2 or 3 substituents.

[0025] In certain embodiments, Ring A is phenyl, optionally substituted with 1, 2, or 3 substituents.

[0026] In certain embodiments, the substituents of ring A are selected from the group consisting of hydrogen, halogen, cyano, nitro, hydroxy, 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;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 1-6Alkyl, -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.

[0027] 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.

[0028] In certain embodiments, R 3 、R 4 are each independently selected from hydrogen, C 1-6 alkyl.

[0029] 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, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo-substituted C1-C6 alkyl, and C1-C6 alkoxy.

[0030] In certain embodiments, the substituents of ring A are selected from the group consisting of hydrogen, halogen, nitro, cyano, hydroxy, 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 fEach 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;

[0031] 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;

[0032] 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 heterocyclyl), -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.

[0033] In certain embodiments, Ring B is a bicyclic heteroaryl.

[0034] In certain embodiments, the ring B is a 5-6 membered monoheteroaryl group containing 1, 2 or 3 nitrogen atoms.

[0035] In certain embodiments, the 5-6 membered monoheteroaryl group containing 1, 2 or 3 nitrogen atoms is selected from the following heteroaromatic ring-forming groups:

[0036] In certain embodiments, Ring B is Optionally, Ring B is substituted with 1, 2 or 3 substituents.

[0037] In certain embodiments, Ring B is Optionally, Ring B is substituted with 1, 2 or 3 substituents.

[0038] In certain embodiments, the substituents of ring B are selected from the group consisting of hydrogen, halogen, nitro, nitroso, cyano, hydroxy, 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, 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 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.

[0039] In certain embodiments, the substituents of ring B are selected from the group consisting of hydrogen, halogen, nitro, nitroso, cyano, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo-C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)Rb’ 、 -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 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, halo-substituted C1-C6 alkyl, C1-C6 alkoxy;

[0040] In certain embodiments, the substituents of Ring B are selected from the group consisting of hydrogen, halogen, nitro, cyano, hydroxy, 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’ .

[0041] 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.

[0042] In certain embodiments, R 3’ 、R 4’ Selected from hydrogen, C1-C6 alkyl.

[0043] In certain embodiments, the 3-6 membered heterocyclyl is substituted by one or more groups selected from the group consisting of halogen, nitro, cyano, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, and C1-C6 alkoxy.

[0044] In certain embodiments, the substituents of Ring B are selected from hydrogen, =0, and C1-C6 alkyl.

[0045] In certain embodiments, Ring B is a pyridone or a pyridazinone.

[0046] In certain embodiments, Ring B is 1H-pyridin-2-one, having the structural formula:

[0047] In certain embodiments, Ring B is 2H-pyridazin-3-one, having the structural formula:

[0048] 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.

[0049] 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-.

[0050] In certain embodiments, L is selected from the following structures: -O-CH2-, -CH2-O-, -NH-CH2-, -CH2-NH-, -CH2-CH2-, -N(Boc)-CH2-.

[0051] In certain embodiments, L is selected from the following structures: -NH-CH2-, -CH2-CH2-, -N(Boc)-CH2-.

[0052] In certain embodiments, q is 0.

[0053] In certain embodiments, T is selected from -(CH-T 1 ) p -, wherein p is selected from an integer between 1 and 6 (eg, 0, 1, 2, 3, 4, 5, or 6), T 1 Selected from: hydrogen, halogen, nitro, hydroxy, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, mercapto, -S-(C1-C6 alkyl).

[0054] In certain embodiments, T 1 Selected from hydrogen, C1-C6 alkyl.

[0055] In certain embodiments, T is selected from -CH2-.

[0056] In certain embodiments, T is -CH(R 2 )-,R 2 Selected from hydrogen or C1-C6 alkyl.

[0057] 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 above, respectively.

[0058] R 2 Selected from hydrogen or C1-C6 alkyl.

[0059] In certain embodiments, R 2 For hydrogen.

[0060] In certain embodiments, R 2 It is a methyl group.

[0061] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (II):

[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 a methyl group.

[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, R7 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] R 1 、R 2 , Q, L, R 5 、R 6 、R 7 As defined in any one of the above items respectively.

[0074] 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.

[0075] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (II"):

[0076] R 1 、R 2 , Q, L, R 5 、R 6 、R 7 As defined in any one of the above items respectively.

[0077] 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-NR3 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.

[0078] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (III):

[0079] in,

[0080] R 1 , Q, L, R 5 、R 6 、R 7 as defined in any of the above items respectively;

[0081] R 2 Selected from hydrogen or C1-C6 alkyl.

[0082] In certain embodiments, R 2 For hydrogen.

[0083] In certain embodiments, R 2 It is a methyl group.

[0084] 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 4Each 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.

[0085] 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.

[0086] 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.

[0087] In certain embodiments, R 7 Selected from hydrogen, C1-C6 alkyl (eg methyl).

[0088] In certain embodiments, R 7 For hydrogen.

[0089] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (III'):

[0090] R 1 、R 2 , Q, L, R 5 、R 6 、R 7 As defined in any one of the above items respectively.

[0091] 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.

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

[0093] R 1 、R 2 , Q, L, R 5 、R 6 、R 7 As defined in any one of the above items respectively.

[0094] In certain embodiments, R 5 For hydrogen.

[0095] 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.

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

[0097] Among them, R 1 , Q, L, R 5 、R 6 、R 7 As defined in any of the above;

[0098] X is C or N;

[0099] R 2 Selected from hydrogen or C1-C6 alkyl.

[0100] In certain embodiments, R 1 For hydrogen.

[0101] In certain embodiments, Q is -O-.

[0102] In certain embodiments, R 2 is hydrogen or methyl.

[0103] In certain embodiments, R 2 For hydrogen.

[0104] In certain embodiments, R 2 It is a methyl group.

[0105] 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.

[0106] In certain embodiments, R 5 、R 6 Each is independently selected from the group consisting of: 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 group consisting of: hydrogen, C1-C6 alkyl.

[0107] In certain embodiments, the -CH2-(3-6 membered nitrogen-containing heterocyclyl) is

[0108] In certain embodiments, R 7 Selected from hydrogen, C1-C6 alkyl (eg methyl).

[0109] In certain embodiments, R 7 For hydrogen.

[0110] In certain embodiments, L is -CH2-CH2- or -NH-CH2-.

[0111] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (V'):

[0112] R 1 、R 2 , Q, L, X, R 5 、R 6 、R 7 As defined in any one of the above items respectively.

[0113] In certain embodiments, R 5 For hydrogen.

[0114] 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.

[0115] In certain embodiments, the -CH2-(3-6 membered nitrogen-containing heterocyclyl) is

[0116] In certain embodiments, R 7 Selected from hydrogen, C1-C6 alkyl (eg methyl).

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

[0118] R 1 、R 2 , Q, L, X, R 5 、R 6 、R 7 As defined in any one of the above items respectively.

[0119] In certain embodiments, R 5 For hydrogen.

[0120] 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.

[0121] In certain embodiments, the -CH2-(3-6 membered nitrogen-containing heterocyclyl) is

[0122] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (V'"):

[0123] R 1 、R 2 , Q, L, X, R 5 、R 6 、R 7 As defined in any one of the above items respectively.

[0124] In certain embodiments, R 5 For hydrogen.

[0125] 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.

[0126] In certain embodiments, the -CH2-(3-6 membered nitrogen-containing heterocyclyl) is

[0127] 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.

[0128] In certain embodiments, R 7 Selected from hydrogen or C1-C6 alkyl.

[0129] In certain embodiments, R 7 Selected from hydrogen.

[0130] In certain embodiments, R 7 Selected from methyl.

[0131] The hydroxyl groups and -NH- and -NH2 in the compounds of the present invention may optionally be 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 scope of protection of the present invention.

[0132] The compounds of the present invention may have a structure selected from the group consisting of:

[0133] Pharmaceutical compositions, uses, and disease prevention / treatment methods

[0134] 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.

[0135] The pharmaceutical compositions of the present invention can be formulated into any pharmaceutically acceptable dosage form and administered to patients in need of such treatment by oral, parenteral, rectal, pulmonary, topical, or transdermal routes. For oral administration, conventional solid preparations such as tablets, capsules, pills, and granules can be prepared; oral liquid preparations such as oral solutions, oral suspensions, and syrups can also be prepared. Suitable fillers, binders, disintegrants, lubricants, and the like can be added to oral preparations. For parenteral administration, injections can be prepared, including injection solutions, sterile powders for injection, and concentrated solutions for injection. Injections can be produced using conventional methods in the pharmaceutical field. Additives may be omitted or added depending on the properties of the drug. For rectal administration, suppositories can be prepared. For pulmonary administration, inhalants or sprays can be prepared. For topical administration, specific dosage forms and devices can be used, or creams, gels, and the like can be prepared. For transdermal administration, patches can be prepared.

[0136] In one aspect, the present application provides the use of a compound of the present invention, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite thereof in the preparation of a medicament, which is used as a TLR7 agonist for preventing or treating a disease associated with TLR7.

[0137] In one aspect, the present application provides a method for preventing or treating a TLR7-related disease in a subject, 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, isotopically labeled form thereof, or a metabolite thereof.

[0138] In some embodiments, the disease associated with TLR7 is selected from:

[0139] (1) tumors, including but not limited to: breast cancer, melanoma, meningioma, soft tissue sarcoma, salivary gland tumor, intraspinal tumor, mediastinal tumor, brain cancer, bone cancer, penile cancer, osteosarcoma, intracranial tumor, tongue cancer, maxillary sinus cancer, thyroid cancer, malignant lymphoma, multiple myeloma, pituitary adenoma, testicular tumor, 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;

[0140] (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;

[0141] (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.

[0142] In the method, the compound of the present invention acts as a TLR7 agonist, and further, can act as an immunomodulator.

[0143] 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 metabolites thereof as vaccine adjuvants. In certain embodiments, the vaccine is used to prevent or treat diseases associated with any of the viral infections described above.

[0144] In one aspect, the present application provides use of a compound of the present invention, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance, or a metabolite thereof as a photodynamic therapy agent.

[0145] In one aspect, the present application provides the use of a compound of the present invention, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystalline form, isotope label or metabolite thereof as a conjugate drug (e.g., an antibody immunostimulatory drug conjugate ISAC). An antibody immunostimulatory drug conjugate (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 Phase II clinical trials for the treatment of metastatic breast cancer.

[0146] In the present application, the subject is preferably a mammal, such as bovine, equine, porcine, canine, feline, rodent, primate, such as human.

[0147] Preparation method

[0148] Route 1:

[0149] Intermediate 1-1 is alkylated with intermediate 1-2 under alkaline conditions to generate 1-3, and intermediate 1-3 completes cyclization through nucleophilic attack under alkaline conditions to obtain the target product 1-4.

[0150] Among them, -X 1 Selected from -OTs, -X 2 is selected from -OTBS, -COOMe, and -CHO, and the remaining substituents are as described above.

[0151] Route 2:

[0152] Intermediate 2-1 is alkylated with intermediate 2-2 under alkaline conditions to generate 2-3. Intermediate 2-3 completes cyclization under alkaline conditions through nucleophilic attack to obtain 2-4. 2-4 undergoes a series of reactions to generate the target product 2-5.

[0153] Among them, -X 1 Selected from -OTs, -X 2 is selected from -OTBS, -COOMe, and -CHO, and the remaining substituents are as described above.

[0154] Definition of terms

[0155] Unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, the laboratory procedures described herein are conventional procedures widely used in the relevant fields. To facilitate a better understanding of the present invention, definitions and explanations of relevant terms are provided below.

[0156] As used herein, the term "C1-C6 alkyl" refers to a group obtained by removing a hydrogen atom from a straight-chain 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, and n-hexyl. In the present invention, the preferred C1-C6 alkyl group is a C1-C4 alkyl group.

[0157] As used herein, the term "C2-C6 alkenyl" refers to a straight-chain 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 group is C2-C4 alkenyl.

[0158] 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 to 6 carbon atoms, specific examples of which 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. In the present invention, the preferred C2-C6 alkynyl group is a C2-C4 alkynyl group. As used herein, the term "halogen" includes fluorine, chlorine, bromine and iodine. In the present invention, the preferred halogen group is fluorine, chlorine, bromine, and more preferably bromine.

[0159] As used herein, the term "halogenated" refers to a group or compound in which hydrogen is replaced by one or more halogen atoms, including full halogenation and partial halogenation.

[0160] As used herein, the term "alkoxy" refers to a group formed in the form of alkyl-O-.

[0161] As used herein, the term "5-6 membered heteroaryl" refers to a monocyclic aromatic group 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.

[0162] As used herein, the term "bicyclic heteroaryl" refers to a bicyclic group in which two rings (at least one of which has aromatic character) 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, which 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.

[0163] As used herein, the term "cycloalkyl" refers to a saturated or partially saturated monocyclic 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.

[0164] As used herein, the term "heterocyclyl" refers to a radical derived from a saturated or partially saturated monocyclic or condensed ring compound containing 3 to 14 ring atoms and at least one heteroatom (e.g., 1, 2, 3, 4, or 5 heteroatoms) by removing one hydrogen atom. 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 further containing 1 or 2 oxygen atoms or sulfur atoms.

[0165] 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 herein with 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), including but not limited to hydrohalides, such as hydrofluorides, hydrochlorides, hydrobromides, hydroiodides, etc.; inorganic acid salts, such as nitrates, perchlorates, sulfates, phosphates, etc.; lower alkanesulfonates, such as methanesulfonates, trifluoromethanesulfonates, ethanesulfonates, etc.; arylsulfonates, 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.

[0166] As used herein, the term "pharmaceutically acceptable ester" refers to an ester formed between the -COOH group present in the compounds provided herein and a suitable alcohol, or an ester formed between the -OH group present in the compounds provided herein 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 in the presence of an acid or base to produce the corresponding acid or alcohol.

[0167] As used herein, the term "pharmaceutically acceptable amide" refers to an amide formed between -COOH present in the compounds provided herein and a suitable amine, or an amide formed between -NH- or -NH2 present in the compounds provided herein and a suitable acid (e.g., carboxylic acid, such as acetic acid).

[0168] As used herein, the term "solvate" refers to a substance formed by the association of a compound of the present invention with a solvent molecule. The solvent may be an organic solvent (e.g., methanol, ethanol, propanol, acetonitrile, etc.). For example, the compound of the present invention may form an ethanolate with ethanol. The compound of the present invention may also form a hydrate with water.

[0169] As used herein, the term "crystalline form" refers to the crystal structure of a substance. During crystallization, various factors may alter the intramolecular or intermolecular bonding patterns, resulting in different arrangements of molecules or atoms within the crystal lattice space, forming different crystal structures. The compounds of the present invention may exist in a single crystal structure or in multiple crystal structures, i.e., they may exhibit "polymorphism." The compounds of the present invention may exist in different crystalline forms.

[0170] As used herein, the term "stereoisomer" includes conformational isomers and configurational isomers, wherein the configurational isomers primarily include cis-trans isomers and optical isomers. The compounds of the present invention may exist in the form of stereoisomers, and therefore encompass all possible stereoisomeric forms, and any combination or any mixture thereof. For example, a single enantiomer, a single diastereomer, or a mixture thereof. When a compound of the present invention contains an olefin double bond, unless otherwise specified, it includes cis-isomers and trans-isomers, and any combination thereof.

[0171] 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 form.

[0172] As used herein, the term "prodrug" refers to a compound that can be converted into the compound of the present invention by reactions such as oxidation, reduction, hydrolysis, etc. in the subject's body. The prodrug itself may or may not have the biological activity of the compound of formula (I) (e.g., regulating glycolipid metabolism activity, anti-inflammatory activity, antioxidant activity). For example, a compound of formula (I) comprising a hydroxyl or carboxyl group can be administered in the form of an ester, which is hydrolyzed in the body to form a hydroxyl compound or a carboxyl compound. Similarly, a compound of formula (I) comprising an amino group can be acylated, alkylated, or phosphorylated to form, for example, an eicosanoylamino group, an alanylamino group, or a pivaloyloxymethylamino group 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).

[0173] 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 the subject and the 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, flavorings, preservatives, taste-masking agents, etc. For example, non-limiting examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, 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. Agents that maintain osmotic pressure include, but are not limited to, sugars, NaCl and the like. Agents 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.

[0174] As used herein, the term "prevention" refers to the method implemented in order to prevent or delay the occurrence of disease or illness or symptom in a subject. As used herein, the term "treatment" refers to the method implemented in order to obtain beneficial or required clinical results. For the purposes of the present invention, beneficial or required clinical results include but are not limited to, alleviating symptoms, narrowing the scope of the disease, stabilizing (that is, 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 (no matter in part or in whole), no matter it is detectable or undetectable. In addition, "treatment" can also refer to, compared with the expected survival (if not receiving treatment), extending the survival.

[0175] As used herein, the term "effective amount" refers to an amount sufficient to obtain or at least partially obtain the 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, stop, 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 stop the disease and its complications of a patient already suffering from the disease. It is 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

[0176] In the following examples, the structures of the compounds were determined by nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LCMS). -6 The units are given in ppm. NMR measurements were performed on a Bruker AVANCE III 500 MHz NMR instrument using deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), etc. as the solvents, and tetramethylsilane (TMS) as the internal standard.

[0177] Analytical liquid chromatography was used to monitor the reaction system and analyze the purity of key 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 3.5 μm, 4.6 x 150 mm.

[0178] Preparative liquid chromatography was used to separate and purify a small amount of important intermediates and the target product. The preparative liquid chromatography system used was an Agilent 1290 Infinity II (equipped with a 1290 Prep Bin Pump, a Prep ALS / FC injector / collector, and a 1260 VWD detector); a C18 reversed-phase silica column.

[0179] Liquid chromatography-mass spectrometry (LC-MS) was used for routine reaction system analysis and molecular weight determination of the target product. LC-MS system: Waters UPLC H-Class ultra-high performance liquid chromatography-mass spectrometry coupled with QDa; chromatographic column: Waters XBridge BEH C18 1.7 μm, 2.1 x 75 mm.

[0180] Liquid phase mobile phase ratios: Acidic conditions: A: 0.05% formic acid in water, B: acetonitrile; Alkaline conditions: A: 0.05% ammonia in water, B: acetonitrile; Neutral conditions: A: 5mM NH4HCO3 in water, B: acetonitrile. The above data are default mobile phase ratios and can be adjusted appropriately according to the properties of the measured samples during analysis and testing.

[0181] Rapid purification system was used for crude preparation and purification of compounds in conventional reaction systems. The instrument model was Biotage Isolera One, and the normal phase and reverse phase columns used were 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 reversed-phase C18 columns are 40g, 80g, 120g and 330g, and the specific parameters are as follows: 20-45μm,

[0182] 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.

[0183] 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, Aikon, Anaiji, Aladdin, J&K, Bid Pharmaceuticals, Leyan, McLean, Shaoyuan (Accela), Titan (Adamas), Xiens, and Inokane.

[0184] Abbreviations

[0185] Protecting group abbreviations: Bn, benzyl, Boc, tert-butyloxycarbonyl, Cbz, benzyloxycarbonyl, MOM, methoxymethyl, PMB, p-methoxybenzyl, TBDPS, tert-butyldiphenylsilyl, TBS, tert-butyldimethylsilyl, THP, 2-tetrahydropyranyl, Tos, p-toluenesulfonyl

[0186] Reagent Abbreviations: ACN, acetonitrile, AIBN, azobisisobutyronitrile, Boc2O, di-tert-butyl dicarbonate, BPO, dibenzoyl peroxide, tBuONa, sodium tert-butoxide, CAM, cerium ammonium molybdate, sulfuric acid color developer, CDI, N,N'-carbonyldiimidazole, m-CPBA, m-chloroperbenzoic acid, DABCO, triethylenediamine, DCE, 1,2-dichloroethane, DCM, dichloromethane, DEAD, diethyl azodicarboxylate, DIAD, diisopropyl azodicarboxylate, DIBAL-H, diisobutylaluminum hydride, DBU, 1,8-diazobisspiro[5.4.0]undec-7-ene, DHP, 3,4-dihydro-2H-pyran, DIEA, N,N-diisopropylethylamine, DMAP, 4-dimethylaminopyridine, DMF, N,N-dimethylformamide, DMSO, dimethyl sulfoxide, EA, ethyl acetate, HMPA, hexamethylphosphoramide, HATU N,N,N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (LAH), lithium aluminum hydride (m-CPBA), m-chloroperbenzoic acid (NBS), N-bromosuccinimide (PE), petroleum ether (PMA), phosphomolybdic acid (PyBOP), 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (TBAF), tetrabutylammonium fluoride (TBDPSCl), tert-butyldiphenylsilyl chloride (TBBSCl), tert-butyldimethylsilyl chloride (TEA), triethylamine (TFA), trifluoroacetic acid (THF), tetrahydrofuran (TPSCl), 2,4,6-triisopropylbenzenesulfonyl chloride (TsCl), p-toluenesulfonyl chloride (TsOH), p-toluenesulfonic acid

[0187] Example 1: Synthesis of Compound C-1

[0188] Step A: Under nitrogen at 0°C, a solution of borane in tetrahydrofuran (20 mL, 1 M in THF, 20.00 mmol) was added dropwise to 2-(3-(bromomethyl)phenyl)acetic acid (2.30 g, 10.04 mmol) in tetrahydrofuran (30 mL). After the addition was complete, the mixture was warmed to room temperature and stirred for 3 hours. Methanol was added to the reaction solution to quench it, and the reaction solution was concentrated under reduced pressure to obtain a yellow crude product. The crude product was separated on a silica gel column (PE:EA = 85:15) to obtain 2-(3-(bromomethyl)phenyl)ethan-1-ol (2.30 g, 8.56 mmol, 85%) as a pale yellow oil. ESI-MS m / z = 215.0 [M+H] + ;Calcu.=214.0.

[0189] 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 2-(3-(bromomethyl)phenyl)ethan-1-ol (1.47 g, 8.65 mmol) dissolved in N,N-dimethylformamide (20 mL) at room temperature. After the addition, the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a yellow solid. The residue was separated on a silica gel column (PE:EA = 85:15) to obtain 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.

[0190] Step C: To a solution of 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)ethan-1-ol (888 mg, 2.93 mmol) in dichloromethane (15 mL) at room temperature were added triethylamine (1.7 mL, 11.99 mmol), 4-dimethylaminopyridine (32 mg, 0.264 mmol), and p-toluenesulfonyl chloride (1.14 g, 6.00 mmol). After complete addition, the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a yellow solid. The residue was separated on a silica gel column (PE:EA = 85:15) to afford 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.

[0191] Step D: 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, cesium carbonate (574 mg, 1.76 mmol) and methyl 6-oxo-1,6-dihydropyridine-3-carboxylate (224 mg, 1.46 mmol) were added at room temperature. After the addition was complete, the whole mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to yield a brown solid. The residue was separated on a silica gel column (DCM:MeOH = 90:10) to yield methyl 1-(3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl)-6-oxo-1,6-dihydropyridine-3-carboxylate (500 mg, crude) as a white solid. ESI-MS m / z = 439.0 [M+H] + ;Calcu.=438.1.

[0192] Step E: To a solution of methyl 1-(3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl)-6-oxo-1,6-dihydropyridine-3-carboxylate (500 mg, crude) in toluene (25 mL) was added diisobutylaluminum hydride (3 mL, 1 M solution in hexane, 3.00 mmol) at room temperature. After complete addition, the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a yellow residue, which was then separated on a silica gel column (DCM:MeOH = 90:10) to afford 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, 10% yield over two steps) as a yellow solid. ESI-MS m / z = 411.0 [M+H] + ;Calcu.=410.1.

[0193] Step F: To a solution of 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) in acetonitrile (5 mL) was added cesium carbonate (143 mg, 0.438 mmol) and lithium iodide (59 mg, 0.438 mmol) at room temperature. After the addition was complete, the mixture was heated to 110°C and stirred under pressure for 36 hours in a sealed tube. The reaction solution was concentrated under reduced pressure to obtain a white crude product, which was then purified by HPLC. Preparation conditions: Waters XBridge RP18 (19 mm × 250 mm, 5 μm); A: 0.1% formic acid in water, B: MeCN; 18 mL / min; 0-4 min, 5% B; 4-16 min, 5%-70% B; 16-17 min, 70%-100% B; detector, UV 254 nm). 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%). 1 H 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.

[0194] Example 2: Synthesis of Compound C-2

[0195] Step A: In a round-bottom flask, 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) were added in sequence and reacted at room temperature. After 18 h, 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, methyl 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (339 mg, crude). The crude product was used directly in the next step without further separation. ESI-MS m / z = 440.0 [M+H] + ;Calcu.=439.1.

[0196] Step B: To a sealed tube, methyl 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (150 mg, 0.34 mmol) and tetrahydrofuran (20 mL) were added sequentially. The mixture was then placed in an ice-water bath at 0°C. Lithium aluminum hydride (52 mg, 1.36 mmol) was added and allowed to react at 0°C. After 5 h, 20 mL of saturated ammonium chloride solution and 20 mL of a 16 / 4 mixture of dichloromethane and methanol (16 mL dichloromethane + 4 mL methanol) were added. The organic layer was separated and the aqueous layer was extracted with 16 / 4 dichloromethane and methanol ((16 mL dichloromethane + 4 mL methanol) × 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product (57 mg, crude) 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 obtained and used directly in the next step. ESI-MS m / z = 412.0 [M+H] + ;Calcu.=411.1.

[0197] Step C: Add the crude product from the previous step (10 mg, 0.024 mmol), cesium carbonate (79 mg, 0.24 mmol), lithium iodide (40 mg, 0.24 mmol), and acetonitrile (3 mL) sequentially to a sealed tube. After all the mixture is added, tighten the tube seal and place 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 (10 mL), and evaporate the solvent under reduced pressure. The crude product is directly subjected to HPLC preparative 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. C-2, solid product (E)-1, is obtained. 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%). 1 H 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=7.5Hz,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-MS m / z=376.0[M+H] + ;Calcu.=375.1.

[0198] Example 3: Synthesis of Compound C-3 / C-5

[0199] Step A: To a round-bottom flask, add methyl 6-oxo-1,6-dihydropyridazine-3-carboxylate (706 mg, 4.6 mmol), cesium carbonate (1.49 g, 4.6 mmol), N,N-dimethylformamide (45 mL), and 5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenethyl-4-methylbenzenesulfonate (820 mg, 1.5 mmol) in sequence. After addition, 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 give a solid product, methyl 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (578 mg, 1.1 mmol, 72%). ESI-MS m / z = 517.9 [M+H] + ;Calcu.=517.0.

[0200] 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-carboxylate (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 mixture was added, the reaction was continued at 0°C and naturally returned to room temperature. After 48 hours, water (50 mL) and a 9 / 1 dichloromethane / methanol mixture (45 mL dichloromethane + 5 mL methanol) were added. The organic layer was separated and the aqueous layer was extracted with a 9 / 1 dichloromethane / methanol mixture ((45 mL dichloromethane + 5 mL methanol) × 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product 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 isolation. ESI-MS m / z = 489.8 [M+H] + ;Calcu.=489.0.

[0201] Step C: Add 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenethyl)-6-(hydroxymethyl)pyridazin-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) sequentially to a sealed tube. After addition, tighten the tube seal and place in a 110°C oil bath for reaction. After 8 h, remove from the oil bath and return to room temperature. Filter to remove cesium carbonate and potassium iodide, rinse with acetonitrile (20 mL), evaporate the solvent under reduced pressure, and submit to preparative HPLC for separation. Preparation conditions: YMC-Actus Triart C18 (20 mm × 250 mm, 5 μm); A: 0.05% ammonia water; B: MeCN; 18 mL / min; 0-3 min, 5% B; 3-4 min, 5%-10% B; 4-24 min, 10%-50% B; detector, UV 254 nm. Solid products C-5 and (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.

[0202] Step D: Replace 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 6After adding all of the following: 2-[(trifluoro-4-boryl)methyl)pyrrolidine-1-one (45 mg, 0.1 mmol), potassium salt of 1-((trifluoro-4-boryl)methyl)pyrrolidine (57 mg, 0.3 mmol), trisdibenzylideneacetone dipalladium (9 mg, 0.01 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (19 mg, 0.04 mmol), and sodium carbonate (32 mg, 0.3 mmol), the atmosphere was replaced with nitrogen three times, and 1,4-dioxane (1.6 mL) and water (0.4 mL) were added sequentially. The Schlenk tube was then placed in an 80°C oil bath for reaction. After 2 hours, the mixture was removed from the oil bath, returned to room temperature, and the solvent was evaporated. The mixture was then separated by preparative HPLC. Preparation conditions: Waters XBridge Prep Shield RP18 (19 mm × 250 mm, 5 μm); A: 0.1% formic acid in water; B: MeCN; 18 mL / min; 0-3 min, 5% B; 3-15 min, 5%-30% B; detector, UV 254 nm. 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%). 1 H 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=5.0 Hz,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.

[0203] Example 4: Synthesis of Compound C-4

[0204] Step A: Replace nitrogen in a dry Schlenk tube three times, then add (E)-1 4 -Amino-3 4 -Br-6 1 ,66 -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 (49 mg, 0.3 mmol), trisdibenzylideneacetone dipalladium (2 mg, 0.002 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (5 mg, 0.01 mmol), and sodium carbonate (32 mg, 0.3 mmol) were added, and the nitrogen atmosphere was replaced three times. 1,4-Dioxane (1.6 mL) and water (0.4 mL) were then added. The Schlenk tube was then placed in an 80°C oil bath for reaction. After 3 h, the mixture was removed from the oil bath, allowed to cool to room temperature, and the solvent was evaporated. The mixture was then separated by preparative HPLC. Preparation conditions: Waters XBridge Prep Shield RP18 (19 mm × 250 mm, 5 μm); A: 0.1% formic acid in water; B: MeCN; 18 mL / min; 5% B from 0 to 3 min; 5% to 30% B from 3 to 23 min; UV detector, 254 nm. The solid product C-4, (E)-1 4 -Amino-3 4 -((dimethylamino)methyl)-6-((dimethylamino)methyl)- 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.

[0205] Example 5: Synthesis of Compound C-6 / C-10

[0206] Step A: To a solution of (2-bromo-5-methylphenyl)acetic acid (15 g, 65.48 mmol) in methanol (100 mL) at room temperature, concentrated sulfuric acid (98%) (0.5 mL, 9.335 mmol) was added. After the addition was complete, the mixture was heated to 80°C and stirred for 6 hours. The reaction solution was concentrated under reduced pressure to obtain a crude pale yellow solid-liquid mixture. The mixture was diluted with water and extracted three times with ethyl acetate (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 the crude yellow oily product, methyl 2-(2-bromo-5-methylphenyl)acetate, which was directly used in the next step. ESI-MS m / z = 244.8 [M+H] + ;Calcu.=244.0.

[0207] Step B: To a solution of crude methyl 2-(2-bromo-5-methylphenyl)acetate and N-bromosuccinimide (11.75 g, 66.02 mmol) in carbon tetrachloride (100 mL) at room temperature, dibenzoyl peroxide (1 g, 4.128 mmol) was added. After complete addition, the mixture was heated to 80°C and stirred for 6 hours. The reaction solution was filtered through silica gel, washed with ethyl acetate, and concentrated under reduced pressure to afford the crude product as a yellow oil. The residue was separated on a silica gel column (SiO2, PE:EA = 90:10). The reaction afforded the product, methyl 2-(2-bromo-5-(bromomethyl)phenyl)acetate (16.28 g, 50.56 mmol, 77% yield over two steps) as a colorless oil.

[0208] Step C: At 0°C under N₂ protection, diisobutylaluminum hydride (126 mL, 126.0 mmol) was added dropwise to methyl 2-(2-bromo-5-(bromomethyl)phenyl)acetate (16.28 g, 50.56 mmol) dissolved in toluene (200 mL). After complete addition, the mixture was stirred at room temperature overnight. The reaction mixture was kept at 0°C, and water was slowly added until no significant bubbles appeared. The mixture was then filtered through celite and extracted three times with ethyl acetate (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 yield the crude product as a colorless oil. The residue was separated on a silica gel column (SiO₂, PE:EA = 67:33). The reaction yielded 2-(2-bromo-5-(bromomethyl)phenyl)ethan-1-ol (9.04 g, 30.75 mmol, 61%) as a white solid. ESI-MS m / z = 274.2 [M-OH] + ;Calcu.=291.9.

[0209] Step D: Potassium carbonate (3.7 g, 26.77 mmol) was added to a solution of 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (3 g, 17.69 mmol) and 2-(2-bromo-5-(bromomethyl)phenyl)ethan-1-ol (7.8 g, 26.53 mmol) in N,N-dimethylformamide (90 mL) and tetrahydrofuran (90 mL) at room temperature. After the addition, the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to afford the crude product as a yellow oil. The residue was purified by silica gel column chromatography (SiO2, DCM:MeOH = 90:10) to afford 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenyl)ethan-1-ol (1.94 g, 5.08 mmol, 29%) as a white solid. ESI-MS m / z=381.8[M+H] + ;Calcu.=381.0.

[0210] Step E: To a solution of 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenyl)ethan-1-ol (1.94 g, 5.08 mmol) and triethylamine (2.8 mL, 20.14 mmol) in dichloromethane (200 mL) at 0°C, p-toluenesulfonyl chloride (1.94 g, 10.18 mmol) was added portionwise. After the addition was complete, the mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to yield a crude yellow oily product. The residue was separated on a silica gel column (SiO2, DCM:MeOH = 10:1) to yield 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%) as a pale yellow solid. ESI-MS m / z = 535.8 [M+H] + ;Calcu.=535.0.

[0211] Step F: To a solution of 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) in N,N-dimethylformamide (10 mL) was added cesium carbonate (911 mg, 2.796 mmol) at room temperature. The mixture was stirred at room temperature overnight. The reaction mixture was filtered through celite, washed with ethyl acetate, and concentrated under reduced pressure to afford the crude product as a brown oil. The residue was separated on a silica gel column (SiO2, PE:EA = 30:70 to EA:MeOH = 90:10). The reaction afforded 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.

[0212] Step G: Tetrabutylammonium fluoride (0.6 mL, 0.60 mmol) was added to a solution of 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) in tetrahydrofuran (20 mL) at room temperature. After complete addition, the mixture was stirred at room temperature overnight. The reaction mixture was filtered through silica gel and washed with ethyl acetate:methanol (50:50). After filtration, the mixture was concentrated under reduced pressure to afford the 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 as a yellow solid, which was used directly in the next step. ESI-MS m / z=488.8[M+H] + ;Calcu.=488.0.

[0213] Step H: To a solution of crude ((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromophenethyl)-5-(hydroxymethyl)pyridin-2(1H)-one in acetonitrile (30 mL) at room temperature, cesium carbonate (790 mg, 2.425 mmol) and lithium iodide (325 mg, 2.428 mmol) were added. After the addition was complete, the mixture was heated to 110°C and stirred for 16 hours. The reaction solution was filtered through celite, washed with ethyl acetate:methanol = 50:50, filtered, and concentrated under reduced pressure to obtain a crude yellow solid product. The residue was separated by TLC (SiO2, ethyl acetate:methanol = 90:10 with 1% triethylamine). The reaction afforded the product (E)-1 as a pale yellow solid. 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%). 10 mg of the product after scraping was subjected to HPLC preparative 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-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 (1.61 mg, 0.003 mmol). 1H NMR (500MHz, DMSO-d6) δppm 8.33(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.

[0214] 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 To a solution of 1,4-dioxane (2 mL) containing 1,4-dioxane (15 mg, 0.033 mmol) and potassium trifluoro(pyrrolidin-1-ylmethyl)borate (38 mg, 0.199 mmol) was added trisdibenzylideneacetone dipalladium (6.1 mg, 0.007 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (12.6 mg, 0.026 mmol), sodium carbonate (21 mg, 0.198 mmol), and water (0.5 mL). After the addition was complete, the mixture was heated to 80°C and stirred under nitrogen for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue. The residue was subjected to HPLC preparative separation under the following 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; 3-18 min, 5%-95% B; 18-19 min, 95%-100% B; detector, UV 254 nm. Compound C-6 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -(pyrrolidin-1-ylmethyl)-6- 1 ,6 2 -Dihydro-1 1H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclosinphen-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.

[0215] Example 6: Synthesis of Compound C-7

[0216] Step A: At room temperature and under N2 protection, (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 2To a solution of 1,4-dioxane (4 mL) containing 2-(morpholin-4-yl)methyl)-1,4-dioxane (25 mg, 0.055 mmol) and potassium (morpholin-4-yl)methyltrifluoroborate (69 mg, 0.288 mmol) was added trisdibenzylideneacetone dipalladium (5 mg, 0.005 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (12 mg, 0.025 mmol), sodium carbonate (35 mg, 0.330 mmol), and water (1 mL). After the addition was complete, the mixture was heated to 80°C and stirred under nitrogen for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was then subjected to HPLC preparative separation under the following conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 18 mL / min; 0-3 min, 10% B; 3-3.5 min, 10%-20% B; 3.5-15 min, 20%-50% B; 15-15.5 min, 50%-100% B; detector, UV 254 nm. Compound C-7 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -(morpholinomethyl)-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)-benzocyclosinphen-6 2 -ketone (2.99 mg, 0.006 mmol, 11%). 1 H 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-MS m / z=474.3[M+H] + ;Calcu.=473.2.

[0217] Example 7: Synthesis of Compound C-8

[0218] Step A: At room temperature and under N2 protection, (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 To a solution of 1,4-dioxane (4 mL) containing 2-nitro-1-oxadiazole (25 mg, 0.055 mmol) and potassium dimethylaminomethyl trifluoroborate (55 mg, 0.288 mmol) was added trisdibenzylideneacetone dipalladium (5 mg, 0.005 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (12 mg, 0.025 mmol), sodium carbonate (35 mg, 0.330 mmol), and water (1 mL). After the addition was complete, the mixture was heated to 80°C and stirred under nitrogen for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was then subjected to HPLC preparative separation under the following conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 18 mL / min; 0-3 min, 10% B; 3-3.5 min, 10%-20% B; 3.5-16 min, 20%-60% B; 16-16.5 min, 60%-100% B; detector, UV 254 nm. Compound C-8 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -((dimethylamino)methyl)-6-((dimethylamino)methyl)- 1 ,6 2 -Dihydro-1 1 H-8-Oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclosinphen-6 2 -ketone (3.43 mg, 0.008 mmol, 14%). 1H 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.

[0219] Example 8: Synthesis of Compound C-9

[0220] Step A: At room temperature and under N2 protection, (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 2To a solution of 1,4-dioxane (4 mL) containing 1-methyl-4-trifluoroboronomethylpiperazine (25 mg, 0.055 mmol) and potassium 1-methyl-4-trifluoroboronomethylpiperazine (73 mg, 0.288 mmol) was added trisdibenzylideneacetone dipalladium (5 mg, 0.005 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (12 mg, 0.025 mmol), sodium carbonate (35 mg, 0.330 mmol), and water (1 mL). After the addition was complete, the mixture was heated to 80°C and stirred under nitrogen for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was then subjected to HPLC preparative separation under the following conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 18 mL / min; 0-3 min, 10% B; 3-3.5 min, 10%-20% B; 3.5-16 min, 20%-50% B; 16-16.5 min, 50%-100% B; detector, UV 254 nm. 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)-benzocyclosinphen-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。

[0221] Example 9: Synthesis of Compound C-11

[0222] Step A: At room temperature, concentrated sulfuric acid (0.5 mL, 9.335 mmol) was added dropwise to a solution of 2-(3-fluoro-5-methylphenyl)acetic acid (5 g, 29.73 mmol) in methanol (50 mL). After complete addition, the mixture was stirred at 100°C for 3 hours. The reaction mixture was neutralized with sodium hydroxide solution and concentrated. Water (50 mL) was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (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 yield the crude product, methyl 2-(3-fluoro-5-methylphenyl)acetate (5.14 g, 28.21 mmol, 95%), as a colorless oil.

[0223] Step B: To a solution of methyl 2-(3-fluoro-5-methylphenyl)acetate (7.88 g, 43.25 mmol) in carbon tetrachloride (75 mL) at room temperature, dibenzoyl peroxide (1.05 g, 4.335 mmol) and N-bromosuccinimide (9.24 g, 51.90 mmol) were added. After complete addition, the 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 on a silica gel column (SiO2, PE:EA = 90:10) to yield methyl 2-(3-(bromomethyl)-5-fluorophenyl)acetate (5.9 g, 22.60 mmol, 52%) as a colorless oil.

[0224] Step C: Under nitrogen at 0°C, diisobutylaluminum hydride (56.5 mL, 56.50 mmol) was added dropwise to methyl 2-(3-(bromomethyl)-5-fluorophenyl)acetate (5.9 g, 22.60 mmol) dissolved in toluene (100 mL). After complete addition, the mixture was stirred at room temperature for 2 hours. Methanol was slowly added to the reaction solution until no bubbles formed. The solid was filtered off, and the organic phase was concentrated under reduced pressure to obtain a white crude product. The crude product was separated on a silica gel column (SiO2, PE:EA = 83:17) to yield 2-(3-(bromomethyl)-5-fluorophenyl)ethan-1-ol (2.5 g, 10.73 mmol, 47%) as a white solid.

[0225] Step D: 2-(3-(Bromomethyl)-5-fluorophenyl)ethan-1-ol (2 g, 8.581 mmol) dissolved in N,N-dimethylformamide (36 mL) was added with 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). The mixture was stirred at room temperature overnight. The reaction mixture was filtered and concentrated under reduced pressure to obtain a crude yellow oil. The crude product was separated on a silica gel column (SiO2, DCM:MeOH = 97:3) to obtain 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenyl)ethan-1-ol (600 mg, 1.865 mmol, 26%) as a yellow solid. ESI-MS m / z=321.9[M+H] + ;Calcu.=321.1.

[0226] Step E: To a solution of 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenyl)ethan-1-ol (600 mg, 1.865 mmol) in dichloromethane (10 mL) at 0°C was added triethylamine (1 mL, 7.459 mmol) and 4-dimethylaminopyridine (22.8 mg, 0.187 mmol), followed by portionwise addition of p-toluenesulfonyl chloride (711 mg, 3.730 mmol). After the addition was complete, the mixture was stirred at room temperature overnight. The mixture was concentrated under reduced pressure to yield the crude product as a yellow oil. The crude product was separated on a silica gel column (SiO2, PE:EA = 1:1) to yield 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenethyl-4-methylbenzenesulfonate (500 mg, 1.051 mmol, 56%) as a pale yellow solid. ESI-MS m / z = 476.2 [M+H] + ;Calcu.=475.1.

[0227] Step F: 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 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenethyl-4-methylbenzenesulfonate (500 mg, 1.051 mmol) dissolved in N,N-dimethylformamide (10 mL) at room temperature. The mixture was stirred overnight at room temperature. The reaction mixture was filtered and concentrated under reduced pressure to obtain a crude yellow oil. The crude product was separated on a silica gel column (SiO2, DCM:MeOH = 90:10). The reaction yielded 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.

[0228] Step G: Tetrabutylammonium fluoride (0.48 mL, 0.48 mmol) was added dropwise to a solution of 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) in tetrahydrofuran (10 mL) at room temperature. The 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 in the next reaction without isolation. ESI-MS m / z = 429.3 [M+H] + ;Calcu.=428.1.

[0229] Step H: Cesium carbonate (228 mg, 0.700 mmol) and lithium iodide (94 mg, 0.70 mmol) were added to a solution of crude 1-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenylethyl)-5-(hydroxymethyl)pyridin-2(1H)-one in acetonitrile (3 mL) at room temperature. The mixture was stirred at 110°C for 16 hours. The mixture was then concentrated under reduced pressure to obtain a yellow crude product, which was then subjected to HPLC preparative separation under the following 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-11 and white solid product (E)-1. 4 -Amino-3 5 -Fluorine-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 cyclosuccinylphen-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.

[0230] Example 10: Synthesis of Compound C-12

[0231] Step A: At room temperature, add azobisisobutyronitrile (1.81 g, 11.01 mmol) to a solution of 1-methoxy-3,5-dimethylbenzene (15 g, 110.1 mmol) and N-bromosuccinimide (20.58 g, 115.6 mmol) in carbon tetrachloride (150 mL). After complete addition, heat the mixture to 80°C and stir for 8 hours. Cool the reaction mixture to room temperature and concentrate under reduced pressure to obtain the crude product. The crude product is separated on a silica gel column (SiO2, 100% PE). This reaction yields 3-(bromomethyl)-1-methoxy-5-toluene (17.25 g, 73%) as a colorless oil. 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).

[0232] Step B: At room temperature, trimethylsilyl cyanide (4.72 g, 47.61 mmol) was added to 3-(bromomethyl)-1-methoxy-5-methylbenzene (5.12 g, 23.80 mmol) and potassium carbonate (4.93 g, 35.71 mmol) dissolved in acetonitrile (50 mL). After complete addition, the mixture was heated to 60°C and stirred for 12 hours. The reaction solution was cooled to room temperature and then made alkaline by adding 1 M sodium hydroxide. The mixture was extracted three times with ethyl acetate (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 the crude product. The crude product was separated on a silica gel column (SiO2, PE:EA = 90:10). This reaction yielded (5-methoxy-3-methylphenyl)acetonitrile (2.52 g, 66%) as a colorless liquid. 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).

[0233] Step C: At room temperature, 3M potassium hydroxide (9.036 mL, 27.11 mmol) was added to (5-methoxy-3-methylphenyl)acetonitrile (1.90 g, 11.79 mmol) dissolved in ethanol (18 mL). After complete addition, the mixture was heated to 80°C and stirred for 12 hours. The reaction mixture was concentrated under reduced pressure and extracted three times with EA (10 mL x 3). The aqueous phase was adjusted to pH 6 with 1M hydrochloric acid and extracted three times with (DCM:MeOH = 6:1) (20 mL x 3). The organic phases were combined, washed once with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield the crude product (5-methoxy-3-methylphenyl)acetic acid (1.60 g, crude). This was used directly in the next reaction.

[0234] Step D: At room temperature, add azobisisobutyronitrile (0.13 g, 0.777 mmol) to a solution of (5-methoxy-3-methylphenyl)acetic acid (1.40 g, 7.769 mmol) and N-bromosuccinimide (1.38 g, 7.769 mmol) in carbon tetrachloride (14 mL). After complete addition, heat the mixture to 100°C and stir for 12 hours. Cool the reaction mixture to room temperature and concentrate under reduced pressure to obtain the crude product. The crude product is separated on a silica gel column (SiO2, DCM:MeOH = 97:3) to obtain (3-(bromomethyl)-5-methoxyphenyl)acetic acid (2.00 g, crude) as a colorless oil. Use it directly in the next reaction.

[0235] Step E: Borane (15.4 mL, 15.4 mmol) was slowly added dropwise to (3-(bromomethyl)-5-methoxyphenyl)acetic acid (2.00 g, 7.72 mmol) dissolved in tetrahydrofuran (20 mL) at 0°C. After the addition, the mixture was slowly warmed to room temperature and stirred overnight. The reaction mixture was placed in an ice bath and quenched by the slow addition of methanol. The mixture was concentrated under reduced pressure to afford the crude product. The crude product was separated on a silica gel column (SiO2, PE:EA = 75:25). This reaction afforded 2-(3-(bromomethyl)-5-methoxyphenyl)ethan-1-ol (550 mg, 22% yield over three steps) as a colorless oil. 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).

[0236] Step F: 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (399 mg, 1.628 mmol) was added to a solution of 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (230 mg, 1.356 mmol) and potassium carbonate (375 mg, 2.713 mmol) in N,N-dimethylformamide (2.5 mL) and tetrahydrofuran (2.5 mL) at room temperature. After the addition, the mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was dissolved in methanol (2 mL) and subjected to reverse phase separation using a Santai SepaFlash C18 (Spherical C18, 20-45 μm, 80g); mobile phase: water (containing 0.05% TFA) and acetonitrile; flow rate: 40 ml / min; gradient: acetonitrile from 0% to 100% over 30 minutes; detection wavelength: 254 nm. Lyophilization under reduced pressure gave 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxyphenyl)ethan-1-ol (80 mg, 16%) as a yellow solid. ESI-MS m / z = 333.9 [M+H] + ;Calcu.=333.1.

[0237] Step G: To a solution of 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxyphenyl)ethan-1-ol (40 mg, 0.12 mmol), triethylamine (0.1 mL, 0.719 mmol), and 4-dimethylaminopyridine (1.46 mg, 0.012 mmol) in dichloromethane (2 mL) at 0°C, p-toluenesulfonyl chloride (91.4 mg, 0.479 mmol) was added. After the addition was complete, the mixture was slowly warmed to room temperature and stirred for 3 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was separated on a silica gel column (SiO2, PE:EA = 75:25). The reaction yielded 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.

[0238] Step H: 6-Oxo-1H-pyridine-3-carbaldehyde (22.7 mg, 0.183 mmol) was added to 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) dissolved in N,N-dimethylformamide (2 mL) at room temperature. After the addition was complete, the mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to give 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) as a white solid. ESI-MS m / z = 438.9 [M+H] + ;Calcu.=438.1.

[0239] Step I: Sodium borohydride (10.3 mg, 0.27 mmol) was added to 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) dissolved in ethanol (2 mL) at 0°C. After the addition, the 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 using a 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 from 0% to 100% over 30 minutes; detection wavelength: 254 nm. 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 (20 mg, 72% yield over two steps), as a white solid. ESI-MS m / z = 440.9 [M+H] + ;Calcu.=440.1.

[0240] Step J: To a solution of 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) in acetonitrile (2 mL) at room temperature was added lithium iodide (65 mg, 0.49 mmol). After the addition was complete, the mixture was heated to 110°C and stirred for 12 hours. The reaction mixture was cooled to room temperature, filtered, and the filter cake was washed with methanol (5 mL). The filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was subjected to HPLC preparative separation under the following conditions: Waters XBridge Prep Shield RP18 (19 mm × 250 mm, 5 μm); A: 5 mM NH4HCO3 in water, B: MeCN; 18 mL / min; 0-3 min, 10% B; 3-3.5 min, 10%-20% B; 3.5-18 min, 20%-80% B; detector, UV 254 nm). The product was then lyophilized under reduced pressure to obtain compound C-12 and a white solid product (E)-1. 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) δppm 8.06 (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.

[0241] Example 11: Synthesis of Compound C-13

[0242] Step A: At 20°C, sulfuric acid (0.65 mL, 12.18 mmol) was slowly added dropwise to a solution of 2-(3,5-dimethylphenyl)acetic acid (20.0 g, 121.8 mmol) in methanol (10 mL). The mixture was then stirred at 70°C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove the excess methanol. Water (50 mL) was then added and the mixture was extracted twice with ethyl acetate (100 mL x 2). The combined organic phases were washed once with saturated sodium chloride (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield a yellow oil. The yellow oil was then separated on a silica gel column (SiO2, PE:EA = 83:17). This yielded methyl 2-(3,5-dimethylphenyl)acetate (21.0 g, 97%) as a yellow oil. 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).

[0243] Step B: At 25°C, add azobisisobutyronitrile (954 mg, 5.813 mmol) to methyl 2-(3,5-dimethylphenyl)acetate (10.36 g, 58.13 mmol) dissolved in 1,2-dichloroethane (100 mL). The mixture is then heated to 80°C. N-bromosuccinimide (20.69 g, 116.26 mmol) is then slowly added portionwise at 80°C. After complete addition, the mixture is heated to 90°C and stirred at 90°C for 0.5 hours. The reaction mixture is cooled to room temperature, and dichloromethane (100 mL) is added. The mixture is then washed once with saturated sodium bicarbonate (100 mL) and once with saturated sodium chloride (100 mL). The mixture is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield a solid residue. The solid residue is then separated on a silica gel column (SiO2, PE:EA = 83:17). The reaction afforded methyl 2-(3,5-bis(bromomethyl)phenyl)acetate (10.15 g, 52%) as a light yellow transparent oily product. 1 H NMR (500MHz, DMSO-d6) δppm 7.43 (s, 1H), 7.29 (d, J = 1.4Hz, 2H), 4.69 (s, 4H), 3.70 (s, 2H), 3.62 (s, 3H).

[0244] Step C: At -30°C under nitrogen, diisobutylaluminum hydride (45.3 mL, 45.3 mmol) was slowly added dropwise to a solution of methyl 2-(3,5-bis(bromomethyl)phenyl)acetate (10.15 g, 30.21 mmol) in dichloromethane (100 mL). After complete addition, the mixture was stirred at 0-5°C under nitrogen for 2 hours. Saturated aqueous ammonium chloride was slowly added to the reaction mixture at 0-5°C until no bubbles formed. After complete addition, the mixture was stirred for 30 minutes, then filtered through celite. The filtrate was collected and the filter cake was washed twice with dichloromethane (200 mL x 2). The collected filtrate was washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a solid residue. The solid residue was separated on a silica gel column (SiO2, PE:EA = 75:25). The reaction afforded bis(bromomethyl)phenyl)ethan-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).

[0245] Step D: At 40°C, 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2.80 g, 16.49 mmol) in N,N-dimethylformamide (3 mL) was slowly added in three portions to 2-(3,5-bis(bromomethyl)phenyl)ethan-1-ol (6.35 g, 20.62 mmol) and potassium carbonate (8.55 g, 61.85 mmol) in N,N-dimethylformamide (3 mL). The mixture was stirred at 40°C for 0.5 h. Water (3 mL) was then added to the mixture, and the mixture was stirred at 40°C for 16 h. The reaction mixture was concentrated under reduced pressure to obtain a solid residue. The solid residue was separated on a silica gel column (SiO2, DCM:MeOH = 94:6). The reaction afforded 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.

[0246] Step E: To a solution of 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) in dichloromethane (5 mL) and methanol (5 mL) at 20°C, manganese dioxide (3.0 g, 34.51 mmol) was slowly added. The mixture was then stirred at 20°C for 7 hours. An additional amount of manganese dioxide (3.0 g, 34.51 mmol) was added to the mixture, and the mixture was stirred at 20°C for an additional 15 hours. The reaction mixture was filtered through celite, and the filter cake was rinsed three times with methanol (20 mL x 3). The filtrate was collected and concentrated under reduced pressure to yield a solid residue. The solid residue was separated on a silica gel column (SiO2, DCM:MeOH = 90:10). The reaction afforded 3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-(2-hydroxyethyl)benzaldehyde (215 mg, 43%) as a white solid product. 1 H NMR(500MHz,DMSO-d6)δppm 9.92(s,1H),8.42(br s,1H),8.18(br s,1H),8.16(s,1H),7.69(br s,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.

[0247] Step F: To a solution of 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) in dichloromethane (5 mL) at 0-5°C, p-toluenesulfonyl chloride (172 mg, 0.904 mmol) was added. The mixture was then stirred at 20°C for 5 hours. The reaction mixture was added to water (20 mL) and extracted twice with dichloromethane (40 mL x 2). The organic phases were combined, washed once with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield a solid residue. The solid residue was separated on a silica gel column (SiO2, DCM:MeOH = 92:8). The reaction yielded 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.

[0248] Step G: To a solution of 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) in N,N-dimethylformamide (5 mL) was added cesium carbonate (188 mg, 0.576 mmol) in N,N-dimethylformamide (5 mL) at 20°C. The mixture was then stirred at 20°C for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain a solid residue. The solid residue was separated on a silica gel column (SiO2, DCM:MeOH = 90:10). The reaction yielded 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.

[0249] Step H: Tetrabutylammonium fluoride (0.046 mL, 0.046 mmol) was slowly added dropwise to a solution of 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) in tetrahydrofuran (5 mL) at 0-5°C. After completion, the mixture was stirred at 20°C for 2 hours. The reaction mixture was added with water (10 mL) and extracted twice with ethyl acetate (20 mL x 2). The organic phases were combined, washed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield a pale yellow solid. The crude product 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). ESI-MS m / z = 438.9 [M+H] + ;Calcu.=438.1.

[0250] Step I: To a solution of 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) and pyrrole (0.007 mL, 0.085 mmol) in dichloromethane (5 mL) at 20°C was added sodium triacetoxyborohydride (18 mg, 0.085 mmol). The mixture was then stirred at 20°C for 18 hours. The reaction mixture was concentrated under reduced pressure to obtain a solid residue. The solid residue was separated on a silica gel column (SiO2, DCM:MeOH = 90:10). The reaction yielded 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) as a yellow oily product. ESI-MS m / z = 494.0 [M+H] + ;Calcu.=493.2.

[0251] Step J: To 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) dissolved in acetonitrile (20 mL) at 20°C were added cesium carbonate (185 mg, 0.57 mmol) and lithium iodide (37.9 mg, 0.28 mmol), and the whole mixture was stirred at 110°C for 2 hours. The reaction solution was filtered through celite, and the filter cake was rinsed three times with ethyl acetate (30 mL * 3). The filtrate was collected and concentrated under reduced pressure to obtain a solid residue, which was subjected to prep-HPLC separation. The preparation conditions were: YMC-Actus Triart 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-(4-amino-2-( ... 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.

[0252] Example 12: Synthesis of Compound C-14

[0253] Step A: At room temperature, sulfuric acid (0.05 mL, 0.83 mmol) was added to a solution of (2-methoxy-5-methylphenyl)acetic acid (1.57 g, 8.71 mmol) in methanol (17 mL). After complete addition, the mixture was heated to 80°C and stirred for 4 hours. After the disappearance of the starting material by TLC, triethylamine solution was added to the system to adjust the pH to neutral. Most of the methanol was evaporated under reduced pressure. Water was added to the concentrate, and the mixture was extracted three times with dichloromethane, dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The reaction yielded the product, methyl (2-methoxy-5-methylphenyl)acetate (1.64 g, 97%), as a yellow oil.

[0254] Step B: At 100°C, add azobisisobutyronitrile (0.14 g, 0.84 mmol) and N-bromosuccinimide (1.81 g, 10.13 mmol) to a solution of methyl (2-methoxy-5-methylphenyl)acetate (1.64 g, 8.44 mmol) in 1,2-dichloroethane (20 mL). After stirring, the reaction mixture was refluxed at 100°C for 7 hours. After cooling to room temperature, 20 mL of water was added. The mixture was extracted with dichloromethane, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. This reaction yielded methyl 2-(5-(bromomethyl)-2-methoxyphenyl)acetate (2.37 g, crude) as a dark brown liquid.

[0255] Step C: To methyl 2-(5-(bromomethyl)-2-methoxyphenyl)acetate (2.31 g, 8.46 mmol) dissolved in N,N-dimethylformamide (190 mL) at room temperature were added 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). After 4 hours of stirring, the starting material disappeared by HPLC. The insoluble material was filtered, the filtrate was adjusted to neutral pH with acetic acid, and then concentrated under reduced pressure. The residue was directly subjected to preparative HPLC separation. Preparation conditions: Welch Xtimate C18 (30 mm × 250 mm, 5 μm); A: 5 mM NH4HCO3 in water; B: MeCN; 40 mL / min, 0-3 min, 10% B; 3-3.5 min, 10%-20% B; 3.5-16 min, 20%-90% B; detector, UV 254 nm. The product, methyl 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenyl)acetate (409 mg, 13% yield over two steps), was obtained as a brown solid. ESI-MS m / z = 361.9 [M+H] + ;Calcu.=361.1.

[0256] Step D: To a solution of methyl 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenyl)acetate (409 mg, 1.13 mmol) in tetrahydrofuran (40 mL) at room temperature was added lithium aluminum hydride (88.0 mg, 2.32 mmol). After complete addition, the mixture was stirred at room temperature for 2 h. The starting material disappeared by TLC. The reaction was quenched by adding water and the pH was adjusted to neutral with dilute hydrochloric acid. Ethyl acetate was added to the mixture, filtered, and the filtrate separated. The aqueous phase was extracted twice with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. This reaction afforded the product, 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenyl)ethan-1-ol (313 mg, 83%), as a brownish-yellow solid.

[0257] Step E: To a solution of 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxyphenyl)ethan-1-ol (313 mg, 0.938 mmol) in dichloromethane (35 mL) at room temperature was added triethylamine (0.52 mL, 3.75 mmol), 4-dimethylaminopyridine (11.5 mg, 0.094 mmol), and p-toluenesulfonyl chloride (357 mg, 1.88 mmol). The mixture was stirred at room temperature overnight. The starting material disappeared after TLC analysis. The reaction mixture was concentrated under reduced pressure, and the residue was separated on a silica gel column (DCM:MeOH = 95:5). The reaction afforded 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.

[0258] 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 sequentially at room temperature, and the mixture was stirred for 5 days. The reaction was monitored by HPLC. After the disappearance of the starting material, the reaction solution was filtered and the pH was adjusted to neutral by adding acetic acid. The N,N-dimethylformamide was evaporated to dryness under reduced pressure, and the residue was separated on a silica gel column (DCM:MeOH=95:5). The reaction afforded 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, 46%) as a white solid. ESI-MS m / z=555.1 [M+H] + ;Calcu.=554.2.

[0259] 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) 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 on a silica gel column (DCM:MeOH = 95:5). The reaction afforded 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%) as a white solid. ESI-MS m / z = 440.9 [M+H] + ;Calcu.=440.1.

[0260] Step H: To 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) dissolved in acetonitrile (30 ml) was added cesium carbonate (850 mg, 2.61 mmol) and lithium iodide (350 mg, 2.61 mmol). The reaction mixture was heated at 110°C in a sealed tube for 10 hours and monitored by HPLC. After the starting material disappeared, the reaction mixture was cooled and the solvent was evaporated under reduced pressure at 35°C. The residue was directly subjected to HPLC preparative 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, UV 254nm. This reaction yielded 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.

[0261] Example 13: Synthesis of Compound C-15

[0262] Step A: To a solution of 5-bromo-1,3-dimethylbenzene (30 g, 162.1 mmol) and azobisisobutyronitrile (2.7 g, 16.21 mmol) in carbon tetrachloride (300 mL) at room temperature, add N-bromosuccinimide (28.9 g, 162.1 mmol). After complete addition, the mixture is stirred at 90°C for 3 hours. The reaction mixture is filtered through celite and concentrated under reduced pressure to yield a yellow crude product. The crude product is separated on a silica gel column (SiO2, PE:EA = 95:5) to yield 5-bromo-3-(bromomethyl)-1-methylbenzene (27.4 g, 64%) as a colorless oil.

[0263] Step B: To a solution of 5-bromo-3-(bromomethyl)-1-methylbenzene (27.4 g, 103.8 mmol) and potassium carbonate (43.0 g, 311.4 mmol) in acetonitrile (300 mL) at room temperature, trimethylsilyl cyanide (20.60 g, 207.6 mmol) was added. The mixture was then heated to 60°C and stirred overnight. The reaction mixture was filtered and concentrated under reduced pressure to afford a crude yellow oil. The crude product was separated on a silica gel column (SiO2, PE:EA = 95:5) to afford (3-bromo-5-methylphenyl)acetonitrile (18.4 g, 84%) as a colorless oil.

[0264] Step C: To a solution of (3-bromo-5-methylphenyl)acetonitrile (18.4 g, 87.59 mmol) in water (50 mL) and ethanol (100 mL) at room temperature, potassium hydroxide (14.74 g, 262.8 mmol) was added. After complete addition, the mixture was heated to 90°C and stirred overnight. The reaction solution was cooled to room temperature, the ethanol was evaporated, and water (50 mL) was added. Dilute hydrochloric acid was then added dropwise to adjust the pH to 1-2. A large amount of white solid precipitated, which was filtered to obtain (3-bromo-5-methylphenyl)acetic acid (19.2 g, 96%) as a white solid.

[0265] Step D: To a solution of (3-bromo-5-methylphenyl)acetic acid (19.24 g, 83.99 mmol) and azobisisobutyronitrile (1.38 g, 8.399 mmol) in carbon tetrachloride (300 mL) at room temperature, add N-bromosuccinimide (14.95 g, 83.99 mmol). After complete addition, the mixture is heated to 80°C and stirred overnight. The reaction mixture is filtered and concentrated under reduced pressure to yield the crude product, 2-(3-bromo-5-(bromomethyl)phenyl)acetic acid, as a pale yellow oil.

[0266] Step E: At 0°C under N2, a 1M solution of borane in tetrahydrofuran (14 g, 168.0 mmol) was added to 200 mL of tetrahydrofuran (containing the crude 2-(3-bromo-5-(bromomethyl)phenyl)acetic acid from the previous step. After complete addition, the mixture was stirred at room temperature under nitrogen for 3 hours. The reaction mixture was kept at 0°C and quenched by slowly adding methanol until no significant bubbles appeared. The temperature was maintained at 10°C during quenching. The mixture was then filtered through celite and concentrated under reduced pressure to yield a yellow crude product. The crude product was separated on a silica gel column (SiO2, PE:EA = 90:10) to yield 2-(3-bromo-5-(bromomethyl)phenyl)ethan-1-ol (8.7 g, 35% yield over two steps) as a white solid.

[0267] Step F: 6-Chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (4.6 g, 27.07 mmol) was added to 2-(3-bromo-5-(bromomethyl)phenyl)ethan-1-ol (8.7 g, 29.59 mmol) and cesium carbonate (13.2 g, 40.61 mmol) dissolved in N,N-dimethylformamide (140 mL) at room temperature. After complete addition, the mixture was stirred at room temperature overnight. The reaction solution was filtered and concentrated under reduced pressure to obtain a crude yellow oil. The crude product was separated on a silica gel column (SiO2, DCM:MeOH = 95:5) to obtain 2-(3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromophenyl)ethan-1-ol (3 g, 27%) as a pale yellow solid. ESI-MS m / z=383.9[M+H] + ;Calcu.=383.0.

[0268] Step G: At room temperature, 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) were dissolved in dichloromethane (100 mL), and triethylamine (4.36 mL, 31.36 mmol) and 4-dimethylaminopyridine (60 mL) were added. After the addition of 4-nitro-1,2-dimethyl-2-thiazolyl-1,2-dimethyl-1 ... + ;Calcu.=537.0.

[0269] Step H: To a solution of 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) in N,N-dimethylformamide (20 mL) at room temperature was added cesium carbonate (2.09 g, 6.427 mmol). After the addition was complete, the mixture was stirred at room temperature overnight. The product was then concentrated under reduced pressure to afford a yellow crude product. The residue was dissolved in N,N-dimethylformamide (15 mL) and separated by reverse phase chromatography using an X-select C18 column (19 mm x 150 mm); mobile phase: water (containing 0.1% formic acid) and acetonitrile; flow rate: 25 mL / min; gradient: acetonitrile from 0% to 100% over 30 minutes; detection wavelength: 254 nm. Freeze-dried under reduced pressure to afford the product, 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, 19%), as a pale yellow solid. ESI-MS m / z = 488.8 [M+H] + ;Calcu.=488.0.

[0270] Step I: At 0°C, sodium borohydride (77.6 mg, 2.05 mmol) was added to a solution of 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) in tetrahydrofuran (40 mL). After complete addition, the mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under reduced pressure to afford 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.

[0271] Step J: To a solution of the crude product from the previous step, 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) in acetonitrile (60 mL) was added lithium iodide (1.1 g, 8.201 mmol) at room temperature. The mixture was stirred at 110°C for 16 hours and then concentrated under reduced pressure to obtain a yellow crude product. The crude product was subjected to preparative HPLC separation using the following 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.

[0272] Example 14: Synthesis of Compound C-16

[0273] 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-62 To a solution of 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (10.5 mg, 0.022 mmol), sodium carbonate (36.3 mg, 0.343 mmol) in water (1 mL) and dioxane (4 mL) was added tris(dibenzylideneacetone)dipalladium (5.1 mg, 0.006 mmol). After complete addition, the mixture was heated to 80°C and stirred under nitrogen for 3 hours. The mixture was then concentrated under reduced pressure to yield a yellow crude product. The crude product was subjected to preparative HPLC separation using the following 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; UV detector, 254 nm. The reaction yielded compound C-16, a white solid product (E)-1 4 -Amino-3 5 -((dimethylamino)methyl)-6-((dimethylamino)methyl)- 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.

[0274] Example 15: Synthesis of Compound C-17

[0275] Step A: At room temperature and under nitrogen protection, dissolve (E)-1 4 -Amino-3 4 -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 To a solution of 1,4-dioxane (10 mL) containing 1,4-dioxane (60 mg, 0.132 mmol) and 1,4-dioxane (10 mL) were added dimethylphosphine oxide (31 mg, 0.397 mmol), tris(dibenzylideneacetone)dipalladium (60 mg, 0.066 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (63 mg, 0.109 mmol), and potassium phosphate (28 mg, 0.396 mmol). After the addition was complete, the mixture was heated to 100°C and stirred under nitrogen for 18 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was then subjected to HPLC preparative separation under the following conditions: Waters XBridge C18 (30 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 40 mL / min; 0-3 min, 5% B; 3-3.5 min, 5%-5% B; 3.5-15 min, 5%-50% B; 15-15.5 min, 50%-100% B; detector, UV 254 nm. 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.

[0276] Example 16: Synthesis of Compound C-18 / M-1

[0277] Step A: To 5-methyl-6-oxo-1,6-dihydropyridine-3-carboxylic acid (919 mg, 6.0 mmol) dissolved in methanol (30 mL) was added thionyl chloride (1.7 mL, 24.0 mmol) at room temperature. After the addition, the mixture was stirred at room temperature and monitored by HPLC until the reaction was complete. The mixture was then concentrated to dryness under reduced pressure. The resulting residue was repeatedly diluted with dichloromethane and concentrated to dryness under reduced pressure. Approximately 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 afforded methyl 5-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (660 mg, 66%) as a brown solid. ESI-MS m / z = 168.0 [M+H] + ;Calcu.=167.1.

[0278] Step B: 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 methyl 5-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (501 mg, 3.0 mmol) were dissolved at room temperature. After the addition was complete, the mixture was stirred at room temperature for 48 hours, filtered, and concentrated under reduced pressure to obtain a solid residue. The residue was separated on a silica gel column (SiO2, DCM:MeOH = 94:6). The reaction afforded the product, methyl 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-carboxylate (340 mg, 43%), as a pale yellow solid. ESI-MS m / z = 532.8 [M+H] + ;Calcu.=532.0.

[0279] Step C: Under nitrogen protection and an ice bath, diisobutylaluminum hydride (3.2 mL, 3.2 mmol) was slowly added to a solution of methyl 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-carboxylate (340 mg, 0.64 mmol) in dichloromethane (32 mL). After complete addition, the mixture was stirred in an ice bath for 1 hour. The reaction was quenched by adding approximately 5 mL of methanol, concentrated to dryness under reduced pressure, and the residue was separated by column chromatography (SiO2, DCM:MeOH = 93:7). The reaction yielded 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.

[0280] 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). Cesium carbonate (780 mg, 2.4 mmol) and lithium iodide (324 mg, 2.4 mmol) were added. After the addition was complete, the mixture was heated to 110°C and stirred for 7 hours. After filtration, the mixture was concentrated under reduced pressure to obtain a solid residue, which was separated by silica gel column (SiO2, EA:MeOH=90:10). The reaction gave M-1 and a pale 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。

[0281] 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. The mixture was filtered, washed with a small amount of methanol, and concentrated to dryness under reduced pressure. The residue was then subjected to HPLC preparative separation (preparative conditions: YMC-Actus Triart C18 (21 mm × 250 mm, 5 μm); A: 0.05% ammonia water, B: MeCN; 18 mL / min; 0-3 min, 5% B; 3-18 min, 5%-60% B; detector, UV 254 nm). The reaction yielded 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(br s,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.

[0282] Example 17: Synthesis of Compound C-19

[0283] Step A: At room temperature and under N2 protection, (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 To a solution of 2-methyltetrahydrofuran (0.5 mL) containing 2-methyltetrahydrofuran (20 mg, 0.044 mmol) of methylsulfonamide (7 mg, 0.074 mmol), allylpalladium(II) chloride dimer (2 mg, 0.005 mmol), 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (7 mg, 0.016 mmol), and potassium carbonate (12 mg, 0.087 mmol) was added. After the addition was complete, the mixture was heated to 80°C and stirred under nitrogen for 18 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was subjected to HPLC preparative separation under the following conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 18 mL / min; 0-3 min, 5% B; 3-3.5 min, 5%-10% B; 3.5-15 min, 10%-50% B; 15-15.5 min, 50%-100% B; detector, UV 254 nm. Compound C-19 was obtained as 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%). 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。

[0284] Example 18: Synthesis of Compound C-20

[0285] 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 To a solution of tris(dibenzylideneacetone)dipalladium (5.1 mg, 0.006 mmol) was added tris(dibenzylideneacetone)dipalladium (5.1 mg, 0.006 mmol) of 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (10.5 mg, 0.022 mmol), sodium carbonate (36.3 mg, 0.343 mmol) in water (1 mL) and dioxane (4 mL). After the addition was complete, the mixture was heated to 80°C and stirred under nitrogen for 3 hours. It was then concentrated under reduced pressure to obtain a yellow crude product. The crude product was subjected to HPLC preparative separation under the following 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). This reaction yielded compound C-20 and a white solid product (E)-1. 4 -Amino-3 5 -(morpholinomethyl)-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 H NMR (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.

[0286] Example 19: Synthesis of Compound C-21

[0287] Step A: At room temperature and under nitrogen protection, (E)-1 was dissolved 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 2To a solution of 2-nitropropane-1-one (20 mg, 0.044 mmol) in dioxane (1 mL) were added 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). After the addition was complete, the mixture was heated to 100°C and stirred under nitrogen for 18 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was then subjected to HPLC preparative separation under the following conditions: Waters XBridge C18 (30 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 40 mL / min; 0-3 min, 5% B; 3-3.5 min, 5%-10% B; 3.5-12 min, 10%-60% B; 12-15 min, 60%-100% B; detector, UV 254 nm. 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%). 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.

[0288] Example 20: Synthesis of Compound C-22 / C-30

[0289] Step A: At room temperature and under nitrogen protection, (E)-1 was dissolved 4 -Amino-3 4 -Br-6 1 ,62 -Dihydro-1 1 H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,5)-pyridine-3(1,3)-benzocyclooctane-6 2 To a solution of 1,1-dioxane (10 mL) containing 1,1'-bis(diphenylphosphinoferrocenedichlorido)-ketone (60 mg, 0.132 mmol) was added tributyl(1-ethoxyethylene)tin (143 mg, 0.396 mmol) and 1,1'-bis(diphenylphosphinoferrocenedichlorido)palladium (97 mg, 0.132 mmol). After complete addition, the mixture was heated to 85°C and stirred under nitrogen for 18 hours. The reaction mixture was concentrated under reduced pressure to yield a brown solid crude residue, which was then subjected to preparative HPLC separation using the following conditions: Waters XBridge C18 (30 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 40 mL / min; 0-3 min, 5% B; 3-3.5 min, 5%-15% B; 3.5-11 min, 15%-60% B; 11-15 min, 60%-100% B; detector: UV 254 nm. Obtain 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%). 1 H 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.

[0290] 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) was dissolved in acetonitrile (containing 1% formic acid, 30 mL), and 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, which was subjected to HPLC preparative separation. Preparation conditions: YMC-Actus Triart C18 (21 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 18 mL / min; 0-3 min, 10% B; 3-3.5 min, 10%-20% B; 3.5-15 min, 20%-80% B; 15-15.5 min, 80%-100% B; detector, UV 254 nm. 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.

[0291] Example 21: Synthesis of Compound C-23 / M-2

[0292] Step A: At room temperature and under nitrogen protection, (E)-1 was dissolved 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 2To a solution of 1,4-dioxane (5 mL) containing 20 mg of bis(1-ethoxyethylene)tin (20 mg, 0.044 mmol) and 1,1'-bis(diphenylphosphinoferrocenedichloropalladium) (32 mg, 0.044 mmol) was added tributyl(1-ethoxyethylene)tin (48 mg, 0.132 mmol). After the addition, the mixture was heated to 85°C and stirred under nitrogen for 18 hours. The reaction mixture was concentrated under reduced pressure to yield a brown solid crude residue, which was then subjected to preparative HPLC separation using the following conditions: Waters XBridge C18 (30 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 40 mL / min; 0-3 min, 5% B; 3-3.5 min, 5%-15% B; 3.5-11 min, 15%-60% B; 11-15 min, 60%-100% B; detector: UV 254 nm. 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.

[0293] Step B: At room temperature, dissolve (E)-1 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, 0.006 mmol) in acetonitrile (containing 1% formic acid, 10 mL), 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, which was subjected to HPLC preparative separation. Preparation conditions: Waters XBridge C18 (30 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 40 mL / min; 0-3 min, 5% B; 3-3.5 min, 5%-15% B; 3.5-11 min, 15%-60% B; 11-15 min, 60%-100% B; detector, UV 254 nm. Compound C-23, a white solid product (E)-3 was obtained. 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.

[0294] Example 22: Synthesis of Compound C-24

[0295] 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 the 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 and ultrasonically dissolved, filtered through a sand core funnel with silica gel and concentrated under reduced pressure. Separation and purification: separation by preparative HPLC. Chromatographic conditions: YMC-Actus Triart C18 (21 mm × 250 mm, 5 μm), A: 0.05% ammonia water, B: ACN; 18 mL / min; 0-3.0 min, 10% B; 3.0-3.5 min, 10%-30% B; 3.5-15 min, 30%-80% B; 15-15.5 min, 80%-100% B; detector, UV 254 nm). After lyophilization, 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.

[0296] Example 23: Synthesis of Compound C-25

[0297] Step A: At room temperature, add (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 (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 the 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) 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 and ultrasonically dissolved, filtered through a sand core funnel with silica gel and concentrated under reduced pressure. Separation and purification: separation by preparative HPLC. Chromatographic conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm), A: 0.05% ammonia, B: ACN; 18 mL / min; 0-3.5 min, 5% B; 3.5-12 min, 5%-50% B; 12-15 min, 50%-80% B (UV detector, 254 nm). After lyophilization, 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.

[0298] Example 24: Synthesis of Compound C-26

[0299] 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 (45 mg, 0.099 mmol), zinc cyanide (18 mg, 0.153 mmol), methanesulfonic acid (2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium (II) (40 mg, 0.050 mmol). After the addition, the reaction system was purged with nitrogen three times. Then, tetrahydrofuran (2.0 mL) and water (2.0 mL) were added, and the mixture was degassed three times to remove oxygen. Thereafter, under a nitrogen atmosphere, the entire mixture was heated to 90°C and stirred for 5 hours. Methanol (20 mL) was added and ultrasonically dissolved, and the mixture was filtered through a sand-core funnel with silica gel and concentrated under reduced pressure. Separation and purification: Separation by preparative HPLC. Chromatographic conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm), A: 0.05% ammonia, B: ACN; 18 mL / min; 0-3.0 min, 10% B; 3.0-17 min, 10%-80% B; 17-18 min, 80%-100% B (UV detector, 254 nm). After lyophilization, compound C-26 and white solid product (E)-1 were obtained. 4 -Amino-6 6 -oxy-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)phenylcyclohexanamide 4 -nitrile (8.85 mg, 21%). 1 H NMR(500MHz,DMSO-d6)δppm 7.94(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.

[0300] Example 25: Synthesis of Compound C-27

[0301] Step A: At room temperature and under nitrogen protection, (E)-1 was dissolved 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 To a solution of 1,4-dimethylphosphine oxide (26 mg, 0.333 mmol), tris(dibenzylideneacetone)dipalladium (50 mg, 0.055 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (64 mg, 0.111 mmol), and potassium phosphate (70 mg, 0.33 mmol) was added to a solution of 1,4-dimethylphosphine oxide (50 mg, 0.11 mmol) in dioxane (10 mL). After the addition was complete, the mixture was heated to 100°C and stirred under nitrogen for 18 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was then subjected to HPLC preparative separation under the following conditions: Welch C18 (30 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 40 mL / min; 0-3 min, 5% B; 3-3.5 min, 5%-10% B; 3.5-15 min, 10%-50% B; 15-15.5 min, 50%-100% B; detector, UV 254 nm. Compound C-27 and white solid product (E)-1 were obtained. 4 -Amino-34 -(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.

[0302] Example 26: Synthesis of Compound C-28 / C-29

[0303] Step A: At room temperature and under nitrogen protection, (E)-1 was dissolved 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 2To a solution of 2-ketone (20 mg, 0.044 mmol) in dimethyl sulfoxide (1 mL) was added cuprous iodide (17 mg, 0.089 mmol) and triethylenediamine (20 mg, 0.178 mmol). After the addition was complete, the mixture was heated to 145°C and stirred under nitrogen for 18 hours. The reaction solution was concentrated under reduced pressure to obtain a brown liquid crude residue, which was subjected to HPLC preparative separation under the following conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 18 mL / min; 0-3 min, 5% B; 3-13 min, 5%-40% B; 13-20 min, 40%-80% B; 20-21 min, 80%-100% B; detector, UV 254 nm. C-29, a 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.

[0304] Example 27: Synthesis of Compound C-31 / C-33

[0305] Step A: At room temperature and under nitrogen protection, (E)-1 was dissolved 4 -Amino-3 4 -Br-6 1 ,6 6 -Dihydro-1 1H-8-oxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(1,3)-pyridazine-3(1,3)-benzocyclooctane-6 6 To a solution of 1,4-dioxane (20 mL) containing 1,4-dioxane (100 mg, 0.220 mmol) of 1,4-dioxane, tributyl(1-ethoxyethylene)tin (238 mg, 0.659 mmol) and 1,1'-bis(diphenylphosphinoferrocenedichloropalladium) (161 mg, 0.220 mmol) were added. After the addition was complete, the mixture was heated to 85°C and stirred under nitrogen for 18 hours. The reaction solution was concentrated under reduced pressure to obtain a brown solid crude residue, which was then subjected to HPLC preparative separation under the following conditions: Welch C18 (30 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 40 mL / min; 0-3 min, 10% B; 3-3.5 min, 10%-20% B; 3.5-15 min, 20%-85% B; 15-15.5 min, 85%-100% B; detector, UV 254 nm. 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%). 1 H NMR(500MHz,DMSO-d6)δppm7.91(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.

[0306] Step B: At room temperature, dissolve (E)-1 4 -Amino-34 -(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 (25 mg, 0.056 mmol) was dissolved in acetonitrile (containing 1% formic acid, 20 mL), and 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, which was subjected to HPLC preparative separation. Preparation conditions: YMC-Actus Triart C18 (21 mm × 250 mm, 5 μm); A: 0.1% formic acid in water, B: MeCN; 18 mL / min; 0-3 min, 10% B; 3-3.5 min, 10%-15% B; 3.5-15 min, 15%-75% B; 15-15.5 min, 75%-100% B; detector, UV 254 nm. Compound C-33, a 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%). 1 H 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.

[0307] Example 28: Synthesis of Compound C-32

[0308] Step A: At room temperature, add (E)-1 4 -Amino-34 -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 the addition, the reaction system was purged with nitrogen three times. Then, 1,4-dioxane (24 mL) and H2O (6 mL) were added, and deoxygenation (degassing) was performed three times to remove oxygen. Then, under a nitrogen atmosphere, the entire mixture was heated to 90°C and stirred for 24 hours. Then, methanol was added for ultrasonic dissolution, filtered, and concentrated under reduced pressure. The resulting mixture was then 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 lyophilization, compound C-32 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -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%). 1H NMR(500MHz,DMSO-d6)δppm 7.89(s,1H),7.79(br s,1H),7.71(s,1H),7.56(br s,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.

[0309] Example 29: Synthesis of Compounds C-34 / C-35 / C-36

[0310] Step A: At room temperature and under nitrogen protection, (E)-1 was dissolved 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 To a solution of 2-ketone (20 mg, 0.044 mmol) in dimethyl sulfoxide (1 mL), cuprous iodide (17 mg, 0.089 mmol) and triethylenediamine (20 mg, 0.178 mmol) were added. After the addition, the mixture was heated to 145°C and stirred under nitrogen for 18 hours. The reaction solution was concentrated under reduced pressure to obtain a brown liquid crude residue, which was subjected to HPLC preparative separation under the following conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 18 mL / min; 0-3 min, 30% B; 3-15 min, 30%-70% B; 15-16 min, 70%-100% B; detector, UV 254 nm. Compound C-34 and white solid product (E)-1 were obtained. 4 -Amino-3 4 -(methylthio)-6- 1 ,6 6 -Dihydro-1 1H-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.

[0311] 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 (40 mg, 0.095 mmol) in dichloromethane (4 mL) was added with m-chloroperbenzoic acid (100 mg, 85% purity, 0.491 mmol). After the addition, the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain a white solid crude residue, which was subjected to HPLC preparative separation. Preparation conditions: Welch C18 (30 mm × 250 mm, 5 μm); A: 0.1% formic acid in water, B: MeCN; 40 mL / min; 0-3 min, 10% B; 3-3.5 min, 10%-15% B; 3.5-15 min, 15%-60% B; 15-15.5 min, 60%-100% B; detector, UV 254 nm. Compound C-35, a white solid product (E)-1, was obtained. 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-66 -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%). 1 H 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.

[0312] Example 30: Synthesis of Compound C-37

[0313] Step A: To a solution of methyl 3-bromo-4-(trifluoromethyl)benzoate (20 g, 70.66 mmol) in tetrahydrofuran (150 mL) at 0°C, add lithium aluminum hydride (2.68 g, 70.66 mmol). After complete addition, the mixture is stirred at room temperature for 3 hours. The reaction mixture is maintained at 0°C, and the quenching agent, methanol, is slowly added until no significant bubbles appear. The temperature is maintained at no more than 10°C during quenching. The mixture is then filtered through celite and concentrated under reduced pressure to obtain the crude product. The crude product is separated on a silica gel column (SiO2, PE:EA = 83:17). This reaction yields (3-bromo-4-(trifluoromethyl)phenyl)methanol (13.89 g, 77%) as a colorless oil.

[0314] Step B: At 0°C, add N-bromosuccinimide (19.39 g, 108.9 mmol) to a solution of (3-bromo-4-(trifluoromethyl)phenyl)methanol (13.89 g, 54.46 mmol) and triphenylphosphine (28.57 g, 108.9 mmol) in dichloromethane (200 mL). After complete addition, stir the mixture at room temperature for 2 hours. Filter the reaction mixture through celite and concentrate under reduced pressure to obtain the crude product. The crude product is separated on a silica gel column (SiO2, PE:EA = 95:5). This reaction yields 2-bromo-4-(bromomethyl)-1-(trifluoromethyl)benzene (13.5 g, 78%) as a colorless oil.

[0315] Step C: To a solution of 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) in N,N-dimethylformamide (300 mL) was added cesium carbonate (20.75 g, 63.69 mmol). The mixture was stirred overnight at room temperature. The reaction mixture was filtered through celite and concentrated under reduced pressure to obtain the crude product. The crude product was separated on a silica gel column (SiO2, DCM:MeOH = 95:5). The reaction yielded 1-(3-bromo-4-(trifluoromethyl)benzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (11.35 g, 66%) as a brown oil. ESI-MS m / z=407.8[M+H] + ;Calcu.=407.0.

[0316] Step D: At room temperature under nitrogen, dichloro(1,1'-bis(diphenylphosphino)ferrocene)palladium (1.02 g, 1.396 mmol) was added to a solution of 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), and (E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.07 g, 5.411 mmol) in H2O (100 mL) and dioxane (400 mL). The mixture was heated to 80°C and stirred overnight under nitrogen. The reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was separated on a silica gel column (SiO2, PE:EA = 50:50). The reaction yielded (E)-6-chloro-1-(3-(2-ethoxyvinyl)-4-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (6.6 g, 59%) as a yellow solid. ESI-MS m / z = 397.9 [M+H] + ;Calcu.=397.1.

[0317] Step E: At 0°C, 48% hydrobromic acid (132 mL) was added to a solution of (E)-6-chloro-1-(3-(2-ethoxyvinyl)-4-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (6.6 g, 16.59 mmol) in water (132 mL) and acetone (132 mL). After complete addition, the mixture was stirred at room temperature overnight. The reaction mixture was kept at 0°C, and then a quenching agent, sodium hydroxide solution, was slowly added until neutral. The temperature was maintained at 10°C during quenching. The mixture was extracted three times with ethyl acetate (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 the crude product. The reaction yielded 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenyl)acetaldehyde (5.4 g, crude product) as a colorless oil. ESI-MS m / z = 369.9 [M+H] + ;Calcu.=369.1.

[0318] Step F: Sodium borohydride (1.38 g, 36.51 mmol) was added to a solution of 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) in tetrahydrofuran (300 mL) at 0°C. After complete addition, the mixture was stirred at room temperature for 2 hours. The reaction mixture was kept at 0°C, and the quenching agent, methanol, was slowly added until no obvious bubbles appeared. The temperature was maintained at 10°C during quenching. The mixture was then filtered through celite and concentrated under reduced pressure to obtain the crude product. The crude product was separated on a silica gel column (SiO2, DCM:MeOH = 90:10). The reaction yielded 2-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenyl)ethan-1-ol (1.7 g, 27% yield over two steps) as a white solid. ESI-MS m / z = 371.9 [M+H] + ;Calcu.=371.1.

[0319] 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 mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure to obtain a brown crude product, which was then separated on a silica gel column (SiO2, DCM:MeOH = 95:5). The reaction afforded a pale 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.

[0320] Step H: To a solution of 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) in N,N-dimethylformamide (40 mL) was added cesium carbonate (372 mg, 1.141 mmol) at room temperature. 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 crude product. Water was added to precipitate a solid, which was filtered and lyophilized to obtain the crude product, methyl 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (309 mg, crude product). ESI-MS m / z = 507.9 [M+H] + ;Calcu.=507.1。

[0321] Step I: At 0°C, sodium borohydride (230 mg, 6.08 mmol) was added to a solution of the crude product from the previous step, methyl 1-(5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-(trifluoromethyl)phenethyl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (309 mg, 0.608 mmol) and calcium chloride (135 mg, 1.216 mmol) in tetrahydrofuran (2 mL) and ethanol (18 mL). The mixture was stirred at room temperature for 3 hours. The reaction mixture was kept at 0°C, and then the quenching agent, methanol, was slowly added until no obvious bubbles appeared. The temperature was kept below 10°C during the quenching process. The mixture was then filtered through celite and concentrated under reduced pressure to obtain the crude product. Water was added to precipitate a solid, which was filtered and lyophilized to give 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). ESI-MS m / z = 479.9 [M+H] + ;Calcu.=479.1.

[0322] Step J: To a solution of 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) in acetonitrile (30 mL) was added cesium carbonate (2.0 g, 6.13 mmol) at room temperature. The mixture was then heated to 110°C and stirred for 8 hours. The mixture was then concentrated under reduced pressure to afford a yellow crude product. The crude product was subjected to preparative HPLC separation using the following 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%). 1 H 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.

[0323] Example 31: Synthesis of Compound C-38

[0324] Step A: To a solution of 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) in N,N-dimethylformamide (30 mL) was added cesium carbonate (279 mg, 0.856 mmol) at room temperature. The mixture was stirred overnight at room temperature and then concentrated under reduced pressure to afford 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。

[0325] Step B: Sodium borohydride (182 mg, 4.82 mmol) was added to a solution of the crude product from 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-dihydropyridine-3-carbaldehyde, in tetrahydrofuran (30 mL) at 0°C. After complete addition, the mixture was stirred at room temperature for 3 hours. The reaction mixture was kept at 0°C, and methanol was slowly added until no obvious bubbles appeared. The temperature was maintained at 10°C during quenching. The mixture was then filtered through celite and concentrated under reduced pressure to obtain a solid residue. The residue was dissolved in N,N-dimethylformamide and separated by reverse phase chromatography using an X select C18 column (19 mm x 150 mm); mobile phase: water (containing 0.1% HCOOH) and acetonitrile; flow rate: 100 ml / min; gradient: acetonitrile from 0% to 50% over 30 minutes; detection wavelength: 254 nm. Lyophilization under reduced pressure afforded the pale 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, 34% yield over two steps). ESI-MS m / z = 479.0 [M+H]+; Calcu. = 478.1.

[0326] Step C: To a solution of 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) in acetonitrile (20 mL) at room temperature, cesium carbonate (633 mg, 1.94 mmol) was added. The mixture was then heated to or cooled to 110°C and stirred for 8 hours. The mixture was then filtered and concentrated under reduced pressure to obtain a yellow crude product. The crude product was subjected to HPLC preparative separation under the following 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-38 and 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-6 2 -ketone (17.80 mg, 21%). 1 H NMR(DMSO-d6,500MHz)δppm 8.30(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.

[0327] Example 32: Synthesis of Compound C-39

[0328] Step A: At room temperature, add (E)-1 4 -Amino-3 4 -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 (19 mg, 0.042 mmol), cyclopropylboronic acid (24 mg, 0.279 mmol), tris(dibenzylideneacetone)dipalladium (14 mg, 0.015 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (30 mg, 0.063 mmol), and potassium carbonate (34 mg, 0.246 mmol) were added. 1,4-dioxane (4.0 mL) and water (1.0 mL) were then added. After stirring, the mixture was degassed three times to remove oxygen from the solvent. The reaction mixture was then heated and stirred at 90°C under nitrogen for 24 hours. The mixture was then filtered through a fritted funnel with a small amount of silica gel, and the solid was washed with methanol (20 mL). The resulting liquid was concentrated under reduced pressure to obtain the crude product, which was then purified by preparative chromatography. Chromatographic conditions were as follows: Welch C18 OBD (30 mm × 250 mm, 5 μm), A: 0.05% Ammonium, 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 lyophilization, 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.

[0329] Example 33: Synthesis of Compound C-40

[0330] 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 To a solution of 1-ketone (100 mg, 0.221 mmol) in dimethyl sulfoxide (5 mL) was added cuprous iodide (84 mg, 0.442 mmol) and triethylenediamine (99 mg, 0.884 mmol). After the addition was complete, the mixture was heated to 145°C and stirred under nitrogen for 18 hours. The reaction solution was concentrated under reduced pressure to obtain a brown liquid crude residue, which was subjected to HPLC preparative separation under the following conditions: Waters XBridge C18 (19 mm × 250 mm, 5 μm); A: 0.05% ammonia in water, B: MeCN; 18 mL / min; 0-3 min, 5% B; 3-3.5 min, 5%-15% B; 3.5-13 min, 15%-15% B; 13-13.5 min, 15%-100% B; detector, UV 254 nm. 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%). 1H 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(br s, 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.

[0331] Example 34: Synthesis of Compound C-41

[0332] Step A: To a solution of (5-amino-3-pyridinyl)methanol (15 g, 120.8 mmol) in dichloromethane (300 mL) was added imidazole (16.45 g, 241.7 mmol) and tert-butyldimethylsilyl chloride (20.03 g, 132.9 mmol, 16.4 mL). Stir at 25°C for 12 hours. Dilute with water (200 mL) and extract with dichloromethane (200 mL x 3). The organic layer was dried over magnesium sulfate, filtered, and concentrated. This afforded 5-(tert-butyl(dimethyl)silyl)oxymethyl)pyridin-3-amine (27.7 g, 96%) 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).

[0333] Step B: To a solution of 5-((tert-butyl(dimethyl)silyl)oxymethyl)pyridin-3-amine (27.7 g, 116.2 mmol) in ethanol (500 mL) was added di-tert-butyl dicarbonate (25.36 g, 116.2 mmol, 26.7 mL). The mixture was stirred at 25°C for 12 hours. The mixture was diluted with water (150 mL) and extracted with ethyl acetate (300 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated. The residue was purified by flash silica gel chromatography ( 330g quick separation The product was purified on a silica gel column (eluent: 0-15% ethyl acetate / petroleum ether gradient @ 100 mL / min) to give tert-butyl N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)carbamate (14.3 g, 36%) as a white solid. 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).

[0334] Step C: To a solution of methyl 4-bromo-3-formylbenzoate (10 g, 41.14 mmol) in methanol (100 mL), sodium borohydride (3.11 g, 82.29 mmol, 2 eq) was added portionwise. 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 x 3), and dried over magnesium sulfate. Filtration and concentration under reduced pressure afforded methyl 4-bromo-3-(hydroxymethyl)benzoate (9.76 g, 39.83 mmol, 97% yield) 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).

[0335] Step D: 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) at 0°C. 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 x 3). The mixture was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to afford methyl 4-bromo-3-(methylsulfonyloxymethyl)benzoate (11.25 g, crude) as a white solid.

[0336] Step E: To N,N-dimethylformamide (100 mL) was added tert-butyl N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)carbamate (4.82 g, 14.23 mmol), followed by the addition of sodium hydride (1.14 g, 28.47 mmol, 60% purity) in portions. The mixture was stirred at 0°C for 30 minutes, followed by the addition of methyl 4-bromo-3-(methylsulfonyloxymethyl)benzoate (4.6 g, 14.23 mmol). The mixture was stirred at 25°C for 2 hours. The mixture was quenched with ice water (100 mL), extracted with ethyl acetate (100 mL*3), and the organic layer was dried over magnesium sulfate, filtered, and concentrated. The residue was purified by flash silica gel chromatography ( 80g quick separation The mixture was purified on a silica gel column with a gradient of 0-15% ethyl acetate / petroleum ether at 100 mL / min as the eluent to afford methyl 4-bromo-3-((tert-butyloxycarbonyl-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridinyl)amino)methyl)benzoate (4.99 g, 52% yield over two steps) as a white solid. 1 HNMR(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。

[0337] Step F: To a solution of methyl 4-bromo-3-((tert-butyloxycarbonyl-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)amino)methyl)benzoate (4.3 g, 7.60 mmol) in tetrahydrofuran (40 mL) was added dropwise diisobutylaluminum hydride (1 M, 38 mL) at 0°C. The mixture was stirred at 25°C for 2 hours. The mixture was diluted with water (50 mL) and 1N dilute hydrochloric acid was added to pH = 7. The mixture was then extracted with ethyl acetate (50 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated to give tert-butyl N-((2-bromo-5-(hydroxymethyl)phenyl)methyl)-N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridyl)carbamate (3.2 g, 78%) as a white solid. ESI-MS m / z = 539.0 [M+H] + ;Calcu.=538.2.

[0338] Step G: To a solution of tert-butyl N-((2-bromo-5-(hydroxymethyl)phenyl)methyl)-N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridinyl)carbamate (3.1 g, 5.77 mmol) in dichloromethane (20 mL) were added triphenylphosphine (1.74 g, 6.63 mmol) and carbon tetrabromide (2.29 g, 6.92 mmol). The mixture was stirred at 25°C for 12 hours. The resulting reaction mixture, tert-butyl N-((2-bromo-5-(bromomethyl)phenyl)methyl)-N-(5-((tert-butyl(dimethyl)silyl)oxymethyl)-3-pyridinyl)carbamate, was used directly in the next step without further treatment or purification. ESI-MS m / z = 601.1 [M+H] + ;Calcu.=600.1.

[0339] Step H: The 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 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) as a yellow oil. ESI-MS m / z = 689.8 [M+H] + ;Calcu.=689.1.

[0340] 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) was added tetrabutylammonium fluoride (1 M, 5.29 mL). The mixture was stirred at 25°C for 2 hours. The mixture was concentrated at 50°C to give 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%) as a white solid. ESI-MS m / z = 575.9 [M+H] + ;Calcu.=575.1。

[0341] 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-pyridyl)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 -Br-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 (400 mg), yellow oil. ESI-MS m / z = 539.8 [M+H] + ;Calcu.=539.1。

[0342] Step K: 1 4 -Amino-3 4 -Br-1 1 Tert-butyl H-8-oxa-5-aza-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(3,5)-pyridine-3(1,3)-benzoate cyclooctane-5-carboxylate (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). The atmosphere was replaced with nitrogen three times and then stirred at 80°C under a nitrogen atmosphere for 12 hours. The reaction mixture was quenched with water (2 mL) and extracted with ethyl acetate (5 mL x 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-tert-butyl 8-oxa-5-aza-1(1,6)-pyrazolo[3,4-d]pyrimidine-6(3,5)-pyridine-3(1,3)-benzocyclooctanecarboxylate (60 mg, crude product). ESI-MS m / z=543.1[M+H]+ ;Calcu.=542.3.

[0343] Step L: Go to 1 4 -Amino-3 4 -(pyrrolidin-1-ylmethyl)-1 1 To 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) were added N,N-diisopropylethylamine (35.7 mg, 276.4 μmol, 48 μL) and trimethylsilyl trifluoromethanesulfonate (61.4 mg, 276.4 μmol). The mixture was stirred at 25°C for 2 hours. The mixture was concentrated in vacuo at 50°C and purified by reverse phase high performance liquid chromatography (column: Max 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(br s,4H). ESI-MS m / z=443.2[M+H] + ;Calcu.=442.2.

[0344] Test Example 1: Cell-level drug efficacy evaluation

[0345] Purpose: Using HEK-Blue TM Evaluation of TLR7 and TLR8 agonist activity of compounds using hTLR7 in vitro cell model

[0346] Experimental Materials:

[0347] 1. Cell line: HEK-Blue TMhTLR7 (InvivoGen), HEK-Blue TM hTLR8 (InvivoGen)

[0348] 2. Reagent: QUANTI-Blue TM Solution (InvivoGen), DMEM (Invitrogen), FBS (HyClone), L-glutamine (GIBCO), TrypLE Express Enzyme (GIBCO), DPBS (GIBCO), Normocin TM (InvivoGen);Penicillin-Streptomycin(GIBCO)

[0349] 3. Instruments: Microplate reader (PerkinElmer, Multimode plate reader 2014), CO2 incubator (THERMO, STERI-CYCLEI160), cell counter (BECKMAN COULTER, vi-cell XR)

[0350] Experimental methods:

[0351] Step 1: Cell plating

[0352] a) Discard the culture medium in the T150 culture flask, add 10 mL of pre-warmed DPBS, and gently shake the culture flask to rinse the cells once. 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.

[0353] b) Add TrypLE TM Discard the Express enzyme and place the tube in the incubator for 1 to 2 minutes.

[0354] c) Add serum-containing medium to terminate digestion, collect the cell suspension, centrifuge at 300 rpm for 5 minutes at room temperature, and discard the supernatant.

[0355] d) Add fresh culture medium, mix the cells by pipetting, and then count the cells using a Vi-Cell XR cell counter.

[0356] e) Dilute the cells to a density of 6.25*10 5 cells / mL, 80 μL of diluted cell suspension was inoculated into a 96-well plate, i.e. 5*10 4 cells / well.

[0357] f) Place the 96-well plate in an incubator and incubate overnight (5% CO 2 , 37° C.).

[0358] Step 2: Drug treatment

[0359] a) The test compound was prepared into a 10 mM stock solution in DMSO;

[0360] b) The test compound 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. DMSO was used as a negative control, and R848 and GS9620 were used as positive controls.

[0361] c) Add 20 μL of the diluted compound to a 96-well plate seeded with 80 μL of cells, diluting the drug 5-fold to a final DMSO concentration of 0.2% in each well. The final concentrations of the test compound and control compound were 20,000, 6,667, 2,222, 741, 247, 82, 27, and 9 nM.

[0362] d) Place the 96-well plate in a 5% CO2, 37°C incubator and incubate for 24 hours.

[0363] Step 3: Activity Detection

[0364] a) Transfer 10 μL of the cell supernatant after compound incubation to a 96-well transparent assay plate, and then add 90 μL of QUANTI-Blue to each well of the assay plate. TM Solution, place in a 5% CO2, 37℃ incubator and incubate for 1 hour.

[0365] b) The absorbance of secreted alkaline phosphatase (SEAP) at 650 nm was detected using a microplate reader.

[0366] Data Analysis:

[0367] The activation fold was calculated using the following formula: activation fold = OD 650 Treatment group / OD 650 GraphPad Prism 7 software was used to fit the curves according to the concentrations of the compounds and the corresponding activation folds, and the EC 50 value.

[0368] Reference substances: R848 (InvivoGen) and GS-9620 (CAYMAN), the structures of which are shown in the figure below:

[0369] The results of hTLR7 activation experiments are shown in the following table:

[0370] The results of the hTLR8 activation experiment of the compound are as follows:

[0371] Test Example 2: Test of the agonistic activity of the compounds of the present invention on human peripheral blood mononuclear cells

[0372] Human peripheral blood mononuclear cells (hPBMCs) represent the primary human immune cell population, including lymphocytes, monocytes, and dendritic cells. These cells express TLR7, TLR8, or TLR9 and respond to stimulation with their corresponding ligands. In this study, hPBMCs were stimulated with potential TLR7 agonists, and levels of IFN-α and TNF-α were measured to reflect the compounds' activation of the TLR7 receptor.

[0373] Experimental Materials:

[0374] 1. Cell line: hPBMCs (Sailybio). Culture conditions: RPMI 1640 medium + 10% (v / v) FBS + 1% Penicillin-Streptomycin, 5% CO2, 37°C.

[0375] 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).

[0376] 3. Instruments: microplate reader (perkinElmer, Multimode plate reader 2014), CO2 incubator (THERMO, STERI-CYCLEI160), cell counter (BECKMAN COULTER, vi-cell XR).

[0377] Experimental methods:

[0378] a) Thaw frozen hPBMCs in a 37°C water bath and transfer to 1640 complete medium. Centrifuge at 500 rpm for 10 minutes at room temperature, discard the supernatant, resuspend the cells in 1640 complete medium, transfer to a culture flask, and culture overnight at 37°C, 5% CO2.

[0379] b) hPBMCs were collected after stable culture, counted, and diluted to 1.11*10 7 cells / mL, 180 μL of diluted cell suspension was inoculated into a 96-well plate, i.e. 2*10 6cells / well, and the 96-well plate was placed in a constant temperature incubator and incubated for 24 h (5% CO2, 37°C).

[0380] 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.

[0381] 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%.

[0382] 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 kit.

[0383] f) Measure the light absorption at 450 nm using a microplate reader, and quantitatively determine the level of corresponding cytokine secretion based on the corresponding standard curve, and calculate the EC by fitting the curve. 50 value.

[0384] g) The experimental results are shown in the following table:

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, hydroxy, 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-, -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 cycloalkane alkyl), -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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