Macrocyclic compound containing polyaryl group and application thereof

Abstract

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

Description

Technical Field

[0001] The present application relates to the field of biomedicine, and in particular to a macrocyclic TLR7 compound containing multiple aromatic groups and uses thereof. Background Art

[0002] Toll-like receptor (TLR) is a pattern recognition receptor expressed on the surface or inside immune cells such as dendritic cells, macrophages, and natural killer cells. Its main function is to activate immune responses and resist infection by recognizing pathogen-associated molecular patterns (PAMPs). There are 10 types of TLRs in humans, and most of them rely on the myeloid differentiation factor 88 (MyD88) pathway for signal transduction. Among them, TLR1 / 2 / 4 / 5 / 6 / 10 are mainly distributed on the cell surface, while TLR3 / 7 / 8 / 9, as nucleotide-sensitive intracellular TLRs, are mainly located inside the cell, synthesized in the endoplasmic reticulum, and eventually transferred to the endosomes.

[0003] TLR7 is a member of the TLR family, mainly expressed in the endosomal membrane of plasmacytoid dendritic cells, and can recognize viral single-stranded RNA (ssRNA), such as ssRNA of HIV and hepatitis C virus. TLR7 can activate the NF-κB signaling pathway and the mitogen-activated protein kinase (MAPK) signaling pathway, and promote the production of inflammatory factors, chemokines and interfering factors. The natural ligand of TLR7 is ssRNA rich in guanosine and / or uridine, and some synthetic heterocyclic small molecules may also have an active regulatory effect on TLR7. TLR7 has three different domains, among which the N-terminal extracellular domain contains 26 leucine-rich repeat sequences (LRR), which form a large parallel β-fold, allowing protein-protein interactions between two monomers. Among them, the unstructured Z-loop region between LRR14 and LRR15 is essential for the dimerization of TLR7.

[0004] TLR7 has been shown to play an important role in the pathogenesis and immune regulation of autoimmune diseases. The TLR7 agonist imiquimod has been approved for the treatment of warts and actinic keratosis caused by papillomavirus. At the same time, the activation of TLR7 can trigger the production of anti-tumor cytokines. Therefore, this type of agonist has also been commonly used in tumor immunotherapy in recent years. Clinical studies have also shown that TLR7 agonists combined with PD-1 inhibitors exhibit good anti-tumor effects. In addition, because of their ability to promote Th1 humoral immune responses, TLR7 agonists are also considered to be an excellent vaccine adjuvant and a potential treatment for allergic asthma.

[0005] In summary, TLR7 plays an important role in the response and activation of the immune system. TLR7 agonists can not only serve as vaccine adjuvants, but also have the potential to fight tumors, fight infections and treat autoimmune diseases, and have great development value. Summary of the invention

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

[0007] Compound

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

[0009]

[0010] in,

[0011] R 1 Selected from: hydrogen, halogen, hydroxyl, amino, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl;

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

[0013] Ring A is selected from phenyl, 5-6 membered heteroaryl, naphthalene ring and bicyclic heteroaryl, wherein the phenyl, 5-6 membered heteroaryl, naphthalene ring and bicyclic heteroaryl ring are optionally substituted by 1, 2 or 3 groups selected from the following: hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, =O, -NR 3 R 4 、-CH 2 R a 、-C(=O)R b 、-OR c, C 3 -C 6 Saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -(C 1 -C 6 Alkyl)OH, -(C 1 -C 6 alkyl)NH 2 、-S(C 1 -C 3 alkyl), -S(=O)(C 1 -C 3 alkyl), -S(=O)(=O)(C 1 -C 3 alkyl), -S(=O)(=O)NH(C 1 -C 3 Alkyl), -NR d S(=O)(=O)(C 1 -C 6 alkyl), -P(=O)R e R f , sulfonic acid, -N=NPh, -Si(C 1 -C 3 alkyl) 3 ; Among them, R a , R b , R c , R d , R e , R f are each independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C 1 -C 6 Alkyl, C 1 -C 3 Alkenyl, C 1 -C 6 Alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH 2 、-NH(C 1 -C 6 Alkyl), -N(C 1 -C 6 alkyl) 2 , C(=O)(C 1 -C 6 alkyl); wherein R 3 , R 4 are each independently selected from hydrogen, C 1-6 Alkyl, C(=O)(C 1 -C 6 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: halogen, nitro, cyano, hydroxyl, C1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy;

[0014] Ring B is selected from phenyl and naphthalene, and the phenyl and naphthalene rings are optionally substituted by 1, 2 or 3 groups selected from the following groups: hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, =O, -NR 3’ R 4’ 、-CH 2 R a’ 、-C(=O)R b’ 、-OR c’ , C 3 -C 6 Saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -(C 1 -C 6 Alkyl)OH, -(C 1 -C 6 alkyl)NH 2 、-S(C 1 -C 3 alkyl), -S(=O)(C 1 -C 3 alkyl), -S(=O)(=O)(C 1 -C 3 alkyl), -S(=O)(=O)NH(C 1 -C 3 Alkyl), -NR d’ S(=O)(=O)(C 1 -C 6 alkyl), -P(=O)R e’ R f’ , sulfonic acid, -N=NPh, -Si(C 1 -C 3 alkyl) 3 ; Among them, R a’ , R b’ , R c’ , Rd’ , R e’ , R f’ are each independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C 1 -C 6 Alkyl, C 1 -C 3 Alkenyl, C 1 -C 6 Alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH 2 、-NH(C 1 -C 6 Alkyl), -N(C 1 -C 6 alkyl) 2 , C(=O)(C 1 -C 6 alkyl); wherein R 3’ , R 4’ are each independently selected from hydrogen, C 1-6 Alkyl, C(=O)(C 1 -C 6 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: halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy;

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

[0016] T is selected from -(CH-T 1 ) p -(Z) q -(CH-T 2 ) r -, wherein Z is selected from -O-, -NH-, -NHC(=O)-, -CH 2 -、-CH(C 1 -C 6 Alkyl)-, -C(C 1 -C 6 Alkyl)(C 1 -C 6 alkyl)-, -C(=O)-, -C(=O)NH-, -N(C 1 -C 6 Alkyl)-, -N(C 1 -C 6 alkyl)C(=O)-, -C(=O)N(C 1 -C 6 alkyl)-, -S-, -S(=O)-, -S(=O)(=O)-, -P(=O)(C 1 -C 6 alkyl)-, -P(=O)(C 1 -C 6 Alkoxy)-, -P(=O)(OH)-, -OC(=O)-, -C(=O)O-, -NHS(=O)(=O)-, -S(=O)(=O)NH-, -N(C 1 -C 6 alkyl)S(=O)(=O)-, -S(=O)(=O)N(C 1 -C 6alkyl)-, p, q, 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, T 1 、T 2 Each is independently selected from: hydrogen, halogen, nitro, hydroxyl, cyano, amino, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, thiol, -S-(C 1 -C 6 alkyl).

[0017] In certain embodiments, R 1 Selected from: hydrogen, halogen, C 1 -C 6 Alkyl, hydroxyl.

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

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

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

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

[0022] In certain embodiments, Ring A is selected from a bicyclic heteroaryl substituted with one or more substituents, such as 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, 1,2,3,4-tetrahydroquinoxaline.

[0023] In certain embodiments, Ring A is selected from a benzene ring, a 5-6 membered heteroaryl, and optionally, Ring A is substituted with 1, 2 or 3 substituents.

[0024] In certain embodiments, the substituents of Ring A are selected from the group consisting of hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C 1-C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, =O, -NR 3 R 4 、-CH 2 R a 、-C(=O)R b 、-OR c ,-SH,-(C 1 -C 6 Alkyl)OH, -(C 1 -C 6 alkyl)NH 2 、-S(C 1 -C 3 alkyl), -S(=O)(C 1 -C 3 alkyl), -S(=O)(=O)(C 1 -C 3 alkyl), -S(=O)(=O)NH(C 1 -C 3 Alkyl), -NR d S(=O)(=O)(C 1 -C 6 alkyl), -P(=O)R e R f , sulfonic acid, -N=NPh, -Si(C 1 -C 3 alkyl) 3 ; Among them, R a , R b , R c , R d , R e , R f are each independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C 1 -C 6 Alkyl, C 1 -C 3 Alkenyl, C 1 -C 6 Alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH 2 、-NH(C 1 -C 6 Alkyl), -N(C 1 -C 6 alkyl) 2 , -C(=O)(C 1 -C 6alkyl); wherein R 3 , R 4 are each independently selected from hydrogen, C 1-6 Alkyl, -C(=O)(C 1 -C 6 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: halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy.

[0025] In certain embodiments, the substituents of Ring A are each independently selected from the following groups: 3 -C 6 Saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, phenyl, 5-6 membered heteroaryl.

[0026] In certain embodiments, the substituents of Ring A are each independently selected from the following groups: hydrogen, halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, =O, -NR 3 R 4 、-CH 2 R a 、-C(=O)R b 、-OR c .

[0027] In certain embodiments, R a , R b , R c , R d , R e , R f are each independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C 1 -C 6 Alkyl, C 1 -C 3 Alkenyl, C 1 -C 6 Alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -N(C1 -C 6 alkyl) 2 Optionally, the 3-6 membered nitrogen-containing heterocyclic group is replaced by one or more C 1 -C 6 Alkyl substituted.

[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 halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy.

[0030] In certain embodiments, Ring A is selected from phenyl, 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are optionally substituted with 1, 2 or 3 groups selected from halogen, C 1 -C 6 Alkyl, halogenated C 1 -C 6 Alkyl, hydroxyl, C 1 -C 6 Alkoxy, =O, -C(=O)H, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 ; Among them, R 3 , R 4 are each independently selected from hydrogen and C 1 -C 6 Alkyl; Optionally, the 3-6 membered nitrogen-containing heterocyclic group is replaced by one or more C 1 -C 6 Alkyl substituted.

[0031] In certain embodiments, the 5-6 membered heteroaryl is a monocyclic heteroaryl, such as a 5-6 membered monocyclic heteroaryl containing nitrogen, oxygen and / or sulfur.

[0032] In certain embodiments, the 5-6 membered heteroaryl group is selected from the following heteroaromatic ring-forming groups:

[0033] In certain embodiments, ring A is selected from an optionally substituted phenyl or an optionally substituted 5-6 membered nitrogen-containing heteroaryl. Optionally, the 5-6 membered nitrogen-containing heteroaryl further contains 1-2 oxygen atoms or sulfur atoms. The 5-6 membered nitrogen-containing heteroaryl may contain 1, 2, 3 or 4 nitrogen atoms, and in certain embodiments, the 5-6 membered nitrogen-containing heteroaryl contains 1 or 2 nitrogen atoms.

[0034] In certain embodiments, the 5-6 membered nitrogen-containing heteroaryl group is selected from the following heteroaromatic ring-forming groups:

[0035] In certain embodiments, Ring A is phenyl, which is optionally substituted with 1, 2 or 3 groups selected from the group consisting of halogen, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), C 1 -C 6 Alkoxy (e.g., methoxy), hydroxy, carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 In certain embodiments, the 3-6 membered nitrogen-containing heterocyclic group is a 5-6 membered heterocyclic group containing 1-2 nitrogen atoms and optionally an oxygen atom.

[0036] In certain embodiments, Ring A is pyridinyl, which is optionally substituted with =0.

[0037] In certain embodiments, Ring B is a naphthyl ring, optionally substituted with 1, 2 or 3 substituents.

[0038] In certain embodiments, Ring B is a benzene ring, optionally substituted with 1, 2 or 3 substituents.

[0039] In certain embodiments, the substituents of Ring B are selected from the group consisting of hydrogen, halogen, nitro, nitroso, cyano, hydroxyl, C 1 -C 6 Alkyl, C2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, =O, -NR 3’ R 4’ 、-CH 2 R a’ 、-C(=O)R b’ 、-OR c’ , C 3 -C 6 Saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -(C 1 -C 6 Alkyl)OH, -(C 1 -C 6 alkyl)NH 2 、-S(C 1 -C 3 alkyl), -S(=O)(C 1 -C 3 alkyl), -S(=O)(=O)(C 1 -C 3 alkyl), -S(=O)(=O)NH(C 1 -C 3 Alkyl), -NR d’ S(=O)(=O)(C 1 -C 6 alkyl), -P(=O)R e’ R f’ , sulfonic acid, -N=NPh, -Si(C 1 -C 3 alkyl) 3 ; Among them, R a’ , R b’ , R c’ , R d’ , R e’ , R f’ are each independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C 1 -C 6 Alkyl, C 1 -C 3 Alkenyl, C 1 -C 6 Alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH 2 、-NH(C 1 -C 6 Alkyl), -N(C 1 -C 6 alkyl) 2, C(=O)(C 1 -C 6 alkyl); wherein R 3’ , R 4’ are each independently selected from hydrogen, C 1-6 Alkyl, C(=O)(C 1 -C 6 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: halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy;

[0040] In certain embodiments, the substituents of Ring B are each independently selected from the following groups: 3 -C 6 Saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, phenyl, 5-6 membered heteroaryl.

[0041] In certain embodiments, the substituents of Ring B are each independently selected from the group consisting of hydrogen, halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, =O, -NR 3’ R 4’ 、-CH 2 R a’ 、-C(=O)R b’ 、-OR c’ .

[0042] In certain embodiments, R a’ , R b’ , R c’ , R d’ , R e’ , R f’ Each independently selected from hydrogen, -OH, C 1 -C 6 Alkyl, C 1 -C 3 Alkenyl, C 1 -C 6Alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -N(C 1 -C 6 alkyl) 2 Optionally, the 3-6 membered nitrogen-containing heterocyclic group is replaced by one or more C 1 -C 6 Alkyl substituted.

[0043] In certain embodiments, R 3’ , R 4’ are each independently selected from hydrogen, C 1-6 alkyl.

[0044] In certain embodiments, the 3-6 membered heterocyclyl is substituted by one or more groups selected from the group consisting of halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy.

[0045] In certain embodiments, Ring B is phenyl, which is optionally substituted with 1, 2 or 3 groups selected from the group consisting of halogen, C 1 -C 6 Alkyl, halogenated C 1 -C 6 Alkyl, hydroxyl, C 1 -C 6 Alkoxy, =O, -C(=O)H, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -NR 3’ R 4’ , carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 ; Among them, R 3’ , R 4’ are each independently selected from hydrogen and C 1 -C 6 Alkyl; Optionally, the 3-6 membered nitrogen-containing heterocyclic group is replaced by one or more C 1 -C 6 Alkyl substituted.

[0046] In certain embodiments, Ring B is phenyl, which is optionally substituted with 1, 2 or 3 groups selected from halogen, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), C 1 -C 6 Alkoxy (e.g., methoxy), hydroxy, carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 In certain embodiments, the 3-6 membered nitrogen-containing heterocyclic group is a 5-6 membered heterocyclic group containing 1-2 nitrogen atoms and optionally an oxygen atom.

[0047] In certain embodiments, Ring B is phenyl, which is optionally substituted with 1, 2 or 3 groups selected from: C 1 -C 6 Alkyl (such as methyl), halogenated C 1 -C 6 Alkyl (e.g. trifluoromethyl), C 1 -C 6 Alkoxy (eg, methoxy).

[0048] In certain embodiments, L is selected from -(CH 2 ) m -W-(CH 2 ) n -, wherein W is selected from -O-, -NH-, -NHC(=O), -CH 2 -、-CH(C 1 -C 6 Alkyl)-, -C(C 1 -C 6 Alkyl)(C 1 -C 6 alkyl)-, -C(=O)-, -C(=O)NH-, -N(C 1 -C 6 and n is independently selected from 0, 1 or 2.

[0049] In certain embodiments, L is selected from the following structures: -O-CH 2-、-NH-CH 2 -、-CH 2 -, -NH-, -NH-C(=O)-, -CH 2 -O-, -CH 2 -O-CH 2 -、-CH 2 -CH 2 -、-O-CH 2 -CH 2 -.

[0050] In certain embodiments, L is selected from the following structures: -O-CH 2 -、-CH 2 -O-, -NH-CH 2 -、-CH 2 -NH-, -CH 2 -CH 2 -.

[0051] In certain embodiments, q is 0.

[0052] In certain embodiments, T is selected from -(CH-T 1 ) p -, wherein p is selected from an integer between 1 and 6 (e.g., 0, 1, 2, 3, 4, 5 or 6), T 1 Selected from: hydrogen, halogen, nitro, hydroxyl, cyano, amino, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, thiol, -S-(C 1 -C 6 alkyl).

[0053] In certain embodiments, T 1 Selected from hydrogen, C 1 -C 6 alkyl.

[0054] In certain embodiments, T is selected from -CH 2 -.

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

[0056]

[0057] R 2 Selected from hydrogen or C 1 -C 6 alkyl.

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

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

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

[0061]

[0062] in,

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

[0064] R 2 Selected from hydrogen or C 1 -C 6 alkyl.

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

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

[0067] R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, =O, -C(=O)H, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -NR 2 R 3 , carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH2 CH=CH 2 ; Among them, R 2 , R 3 are each independently selected from hydrogen and C 1 -C 6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy.

[0068] In certain embodiments, R 5 , R 6 , R 7 Each independently selected from: hydrogen, halogen, C 1 -C 6 Alkyl, halogenated C 1 -C 6 Alkyl, hydroxyl, C 1 -C 6 Alkoxy, =O, -C(=O)H, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 ; Among them, R 3 , R 4 are each independently selected from hydrogen and C 1 -C 6 Alkyl; Optionally, the 3-6 membered nitrogen-containing heterocyclic group is replaced by one or more C 1 -C 6 Alkyl substituted.

[0069] In certain embodiments, R 5 , R 6 are each independently selected from hydrogen, halogen, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), C1 -C 6 Alkoxy (e.g., methoxy), hydroxy, carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 In certain embodiments, the 3-6 membered nitrogen-containing heterocyclic group is a 5-6 membered heterocyclic group containing 1-2 nitrogen atoms and optionally an oxygen atom.

[0070] In certain embodiments, R 7 Selected from hydrogen, C 1 -C 6 Alkyl (such as methyl), halogenated C 1 -C 6 Alkyl (e.g. trifluoromethyl), C 1 -C 6 Alkoxy (eg, methoxy).

[0071] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (II'):

[0072]

[0073] R 1 , R 2 , Q, L, R 5 , R 6 , R 7 As defined in any of the above items respectively.

[0074] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (II"):

[0075]

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

[0077] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (III):

[0078]

[0079] in,

[0080] R 1, Q, L are as defined in any of the above items;

[0081] R 2 Selected from hydrogen or C 1 -C 6 alkyl.

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

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

[0084] R 5 , R 6 Each is independently selected from: hydrogen, halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, =O, -C(=O)H, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 ; Among them, R 3 , R 4 are each independently selected from hydrogen and C 1 -C 6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: hydrogen, halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy.

[0085] In certain embodiments, R5 , R 6 Each independently selected from: hydrogen, halogen, C 1 -C 6 Alkyl, halogenated C 1 -C 6 Alkyl, hydroxyl, C 1 -C 6 Alkoxy, =O, -C(=O)H, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 ; Among them, R 3 , R 4 are each independently selected from hydrogen and C 1 -C 6 Alkyl; Optionally, the 3-6 membered nitrogen-containing heterocyclic group is replaced by one or more C 1 -C 6 In certain embodiments, R 5 , R 6 are each independently selected from hydrogen, halogen, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), C 1 -C 6 Alkoxy (e.g., methoxy), hydroxy, carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 .

[0086] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (III'):

[0087]

[0088] R 1 , R 2 , Q, L, R 5 , R 6 As defined in any of the above items respectively.

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

[0090]

[0091] R 1 , R 2 , Q, L, R 5 , R 6 As defined in any of the above items respectively.

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

[0093]

[0094] in,

[0095] R 1 , Q, L are as defined in any of the above items;

[0096] R 2 Selected from hydrogen or C 1 -C 6 alkyl.

[0097] In certain embodiments, R 2 For hydrogen.

[0098] In certain embodiments, R 2 It is methyl.

[0099] One of X and Y is C or CH, and the other is N or NH;

[0100] R 5 , R 7 Each is independently selected from: hydrogen, halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, =O, -C(=O)H, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C1 -C 6 Alkyl), -OCH 2 CH=CH 2 ; Among them, R 3 , R 4 are each independently selected from hydrogen and C 1 -C 6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy.

[0101] In certain embodiments, R 5 , R 7 Each independently selected from: hydrogen, halogen, C 1 -C 6 Alkyl, halogenated C 1 -C 6 Alkyl, hydroxyl, C 1 -C 6 Alkoxy, =O, -C(=O)H, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 Alkyl), -OCH 2 CH=CH 2 ; Among them, R 3 , R 4 are each independently selected from hydrogen and C 1 -C 6 Alkyl; Optionally, the 3-6 membered nitrogen-containing heterocyclic group is replaced by one or more C 1 -C 6 Alkyl substituted.

[0102] In certain embodiments, R 5 is hydrogen or =O;

[0103] In certain embodiments, R 7 Selected from hydrogen, C 1-C 6 Alkyl (such as methyl), halogenated C 1 -C 6 Alkyl (e.g. trifluoromethyl), C 1 -C 6 Alkoxy (eg, methoxy).

[0104] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (IV'):

[0105]

[0106] R 1 , R 2 , Q, L, R 5 , R 7 As defined in any of the above items respectively.

[0107] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (IV"):

[0108]

[0109] R 1 , R 2 , Q, L, R 5 , R 7 As defined in any of the above items respectively.

[0110] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (V):

[0111]

[0112] Among them, R 1 , Q, L are as defined in any of the above items;

[0113] R 2 Selected from hydrogen or C 1 -C 6 alkyl.

[0114] In certain embodiments, R 2 For hydrogen.

[0115] In certain embodiments, R 2 It is methyl.

[0116] R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2-C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, =O, -C(=O)H, -CH 2 -OH, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C 1 -C 6 Alkyl), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 ; Among them, R 3 , R 4 are each independently selected from hydrogen and C 1 -C 6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the following: halogen, nitro, cyano, hydroxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, halo C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy.

[0117] In certain embodiments, R 1 For hydrogen.

[0118] In certain embodiments, Q is -O-.

[0119] In certain embodiments, R 2 It is hydrogen or methyl.

[0120] In certain embodiments, R 5 Hydrogen, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 .

[0121] In certain embodiments, R 5Methyl, methoxy, halogen, hydroxyl, -COOH, -COOCH 3 、-CH 2 OH. In certain embodiments, R 6 Hydrogen, -CH 2 -(3-6 membered nitrogen-containing heterocyclic group), -CH 2 -N(C 1 -C 6 alkyl) 2 、-OCH 2 CH=CH 2 .

[0122] In certain embodiments, R 6 Methyl, methoxy, halogen, hydroxyl, -COOH, -COOCH 3 、-CH 2 OH.

[0123] In certain embodiments, R 7 For hydrogen, halogen, C 1 -C 6 Alkoxy (eg, methoxy).

[0124] In certain embodiments, L is -CH 2 -O-, -O-CH 2 -.

[0125] In certain embodiments, L is -NHCH 2 -.

[0126] The hydroxyl groups and -NH-, -NH 2 Optionally, it may be protected by a substituent, which may be a hydroxyl or amino protecting group commonly used in the art, such as Boc (tert-butyloxycarbonyl), TBDPS (tert-butyldiphenyl), etc. The compounds thus formed are also within the protection scope of the present invention.

[0127] The compounds of the present invention may have a structure selected from the following:

[0128]

[0129]

[0130]

[0131] Pharmaceutical composition, use, and disease prevention / treatment methods

[0132] 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.

[0133] The pharmaceutical composition of the present invention can be prepared into any pharmaceutically acceptable dosage form and applied to patients who need such treatment by oral, parenteral, rectal, pulmonary, topical or transdermal administration. When used for oral administration, it can be prepared into conventional solid preparations, such as tablets, capsules, pills, granules, etc.; it can also be prepared into oral liquid preparations, such as oral solutions, oral suspensions, syrups, etc. When made into oral preparations, suitable fillers, binders, disintegrants, lubricants, etc. can be added. When used for parenteral administration, it can be prepared into injections, including injections, sterile powders for injection and concentrated solutions for injection. When made into injections, conventional methods in the existing pharmaceutical field can be used for production. When preparing injections, additives may not be added, or suitable additives may be added according to the properties of the drug. When used for rectal administration, it can be prepared into suppositories, etc. When used for pulmonary administration, it can be prepared into inhalants or sprays, etc. When used for topical administration, it can be used in specific dosage forms and devices or prepared into creams, gels, etc. When used for transdermal administration, it can be prepared into patches.

[0134] In one aspect, the present application provides the use of the compounds of the present invention, their pharmaceutically acceptable salts, prodrugs, stereoisomers, tautomers, tautomers, hydrates, solvates, crystal forms, isotope labels or their metabolite forms for preparing drugs, which are used as TLR7 agonists for preventing or treating diseases associated with TLR7.

[0135] In one aspect, the present application provides a method for preventing or treating a TLR7-related disease in a subject, the method comprising administering to a subject in need thereof an effective amount of a compound of the present invention, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystalline form, isotope-labeled substance or a metabolite thereof.

[0136] In some embodiments, the disease associated with TLR7 is selected from:

[0137] (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;

[0138] (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;

[0139] (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.

[0140] In the method, the compound of the present invention acts as a TLR7 agonist, and further, can act as an immunomodulator.

[0141] The compounds of the present invention have an immunoenhancing effect. Therefore, in one aspect, the present application provides the use of the compounds of the present invention, their pharmaceutically acceptable salts, prodrugs, stereoisomers, tautomers, hydrates, solvates, crystal forms, isotope labels or their metabolites as vaccine adjuvants. In certain embodiments, the vaccine is used to prevent or treat diseases associated with any of the viral infections described above.

[0142] In one aspect, the present application provides use of the compound of the present invention, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope label or metabolite form thereof as a photodynamic therapy agent.

[0143] In one aspect, the present application provides the use of a compound of the present invention, a pharmaceutically acceptable salt, a prodrug, a stereoisomer, a tautomer, a hydrate, a solvate, a crystalline form, an isotope label or a metabolite thereof as a conjugated drug (e.g., an antibody immunostimulatory conjugate drug ISAC). An antibody immunostimulatory conjugate drug (ISAC) is a monoclonal antibody that combines an antibody with an immunostimulatory agent to enhance the immune response by triggering the production of proinflammatory cytokines and chemokines. For example, BDC-1001 is an ISAC that includes an antibody targeting HER2 and a TLR7 / 8 agonist, and is currently in clinical phase I / II for the treatment of solid tumors.

[0144] In the present application, the subject is preferably a mammal, such as bovine, equine, porcine, canine, feline, rodent, primate, such as human.

[0145] Preparation method

[0146]

[0147] Intermediate 1-1 and intermediate 1-2 are alkylated under alkaline conditions to generate intermediate 1-3, and then intermediate 1-3 undergoes a series of reactions (including cyclization through nucleophilic attack under alkaline conditions) to obtain the target product 1-4.

[0148] Among them, -X 1 Selected from -CH 2 OH, -CH 2 OTBS, -CH 2 OTBDPS, -CH 2 OAc, -COOMe, -CHO, -X 2 Selected from -Br, -Cl, -OMs, and the remaining substituents are as described above.

[0149] Definition of terms

[0150] In the present invention, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. In addition, the laboratory operation steps involved in this article are all conventional steps widely used in the corresponding fields. At the same time, in order to better understand the present invention, the definitions and explanations of relevant terms are provided below.

[0151] As used herein, the term “C 1 -C 6"Alkyl" refers to a group obtained by removing a hydrogen atom from a straight or branched alkane containing 1 to 6 carbon atoms, and specific examples thereof 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 C 1 -C 6 Alkyl is C 1 -C 4 alkyl.

[0152] As used herein, the term “C 2 -C 6 "Alkenyl" refers to a straight or branched hydrocarbon group containing at least one carbon-carbon double bond and having 2 to 6 carbon atoms, and specific examples thereof 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, 1,4-hexadienyl. In the present invention, preferred C 2 -C 6 Alkenyl is C 2 -C 4 Alkenyl.

[0153] As used herein, the term “C 2 -C 6 "Alkynyl" refers to a straight or branched hydrocarbon group containing at least one triple bond and having 2 to 6 carbon atoms, and specific examples thereof include but are not limited to ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 1,3-butadiynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,4-hexadiynyl. In the present invention, preferred C 2 -C 6 Alkynyl is C 2 -C 4 As used herein, the term "halogen" includes fluorine, chlorine, bromine and iodine. In the present invention, preferred halogen is fluorine, chlorine, bromine, and more preferred is bromine.

[0154] As used herein, the term "halogenated" means that hydrogen on a group or compound is replaced by one or more halogen atoms, including full halogenation and partial halogenation.

[0155] As used herein, the term "alkoxy" refers to a group formed in the form of alkyl-O-.

[0156] As used herein, the term "5-6 membered heteroaryl" refers to a monocyclic group with aromaticity containing 5-6 ring atoms and at least one ring atom being a heteroatom, wherein the heteroatom is a nitrogen atom, an oxygen atom and / or a sulfur atom. Specific examples include, but are not limited to, furyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl.

[0157] As used herein, the term "bicyclic heteroaryl" refers to a bicyclic group in which two rings (at least one of which has aromaticity) are fused together and at least one ring atom is a heteroatom, which is a nitrogen atom, an oxygen atom and / or a sulfur atom, and the nitrogen and / or sulfur are optionally substituted with oxo. Specific examples include, but are not limited to, benzopyrrole, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzimidazole, benzotriazole, indazole, purine, indole, quinoline, pteridine, isoquinoline, quinazoline, quinoxaline, 1,8-naphthyridine, 1,5-naphthyridine, 9H-purine, 7H-purine, indane, indoline, 2,3-dihydrobenzofuran, 1,2,3,4-tetrahydronaphthalene, 1,2,3,4-tetrahydroquinoline, 3,4-dihydro-1H-benzopyran, 1,4-benzodioxane, benzomorpholine, and 1,2,3,4-tetrahydroquinoxaline.

[0158] As used herein, the term "cycloalkyl" refers to a saturated or partially saturated monocyclic cyclic alkyl group that is not aromatic, preferably having 3 to 6 ring atoms, ie, a 3-6-membered saturated cycloalkyl group and a 3-6-membered partially saturated cycloalkyl group. Specific examples of 3-6 membered saturated cycloalkyl groups include, but are not limited to, cyclopropanyl (cyclopropyl), cyclobutanyl (cyclobutyl), cyclopentanyl (cyclopentyl), and cyclohexanyl (cyclohexyl); specific examples of 3-6 membered partially saturated cycloalkyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexa-1,3-dienyl, cyclohex-1,4-dienyl, cycloheptenyl, cyclohept-1,3-dienyl, cyclohept-1,4-dienyl, cyclohept-1,3,5-trienyl, cyclooctenyl, cyclooct-1,3-dienyl, cyclooct-1,4-dienyl, cyclooct-1,5-dienyl, cyclooct-1,3,5-trienyl, and cyclooctatetraenyl.

[0159] As used herein, the term "heterocyclyl" refers to a group obtained by removing a hydrogen atom from a saturated or partially saturated monocyclic or condensed ring compound containing 3-14 ring atoms and at least one heteroatom (e.g., 1, 2, 3, 4 or 5 heteroatoms). The term "3-6 membered nitrogen-containing heterocyclyl" refers to a heterocyclyl containing 3, 4, 5 or 6 ring atoms, 1, 2 or 3 of which are nitrogen atoms, and optionally, the heterocyclyl further contains 1 or 2 oxygen atoms or sulfur atoms.

[0160] As used herein, the term "pharmaceutically acceptable salt" refers to (i) a salt formed by an acidic functional group (e.g., -COOH) present in the compounds provided by the present invention and a suitable inorganic or organic cation (base), and includes, but is not limited to, alkali metal salts, such as sodium salts, potassium salts, lithium salts, etc.; alkaline earth metal salts, such as calcium salts, magnesium salts, etc.; other metal salts, such as aluminum salts, iron salts, zinc salts, copper salts, nickel salts, cobalt salts, etc.; inorganic base salts, such as ammonium salts; organic base salts, such as tert-octylamine salts, dibenzylamine salts, morpholine salts, glucosamine salts, phenylglycine alkyl ester salts, ethylenediamine salts, N-methylglucosamine salts, guanidine salts, diethylamine salts, triethylamine salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, chloroprocaine salts, procaine salts, diethanolamine salts, N-benzyl-phenethylamine salts, piperazine salts, tetramethylamine salts, tris(hydroxymethyl)aminomethane salts. And, (ii) the basic functional groups (e.g. -NH 2 ) and a suitable inorganic or organic anion (acid) to form a salt, and includes but is not limited to, hydrohalide salts, such as hydrofluoride, hydrochloride, hydrobromide, hydroiodide, etc.; inorganic acid salts, such as nitrate, perchlorate, sulfate, phosphate, etc.; lower alkane sulfonate salts, such as methanesulfonate, trifluoromethanesulfonate, ethanesulfonate, etc.; aryl sulfonate salts, such as benzenesulfonate, p-toluenesulfonate, etc.; organic acid salts, such as acetate, malate, fumarate, succinate, citrate, tartrate, oxalate, maleate, etc.; amino acid salts, such as glycine, trimethylglycine, arginine, ornithine, glutamate, aspartate, etc.

[0161] As used herein, the term "pharmaceutically acceptable ester" refers to an ester formed by -COOH present in the compound provided by the present invention and a suitable alcohol, or an ester formed by -OH present in the compound provided by the present invention and a suitable acid (e.g., a carboxylic acid or an oxygen-containing inorganic acid). Suitable ester groups include, but are not limited to, formates, acetates, propionates, butyrates, acrylates, ethylsuccinates, stearic acid esters or palmitates. Esters can undergo hydrolysis reactions to generate corresponding acids or alcohols in the presence of acids or bases.

[0162] As used herein, the term "pharmaceutically acceptable amide" refers to an amide formed by -COOH present in the compounds provided by the present invention and a suitable amine, or -NH- or -NH present in the compounds provided by the present invention. 2 Amides formed with appropriate acids (eg, carboxylic acids such as acetic acid).

[0163] As used herein, the term "solvate" refers to a substance formed by the association of the compounds of the present invention with solvent molecules. The solvent may be an organic solvent (e.g., methanol, ethanol, propanol, acetonitrile, etc.), for example, the compounds of the present invention may form ethanolates with ethanol. The compounds of the present invention may also form hydrates with water.

[0164] As used herein, the term "crystal form" refers to the crystal structure of a substance. When a substance crystallizes, various factors may affect the intramolecular or intermolecular bonding mode, causing molecules or atoms to be arranged differently in the lattice space, forming different crystal structures. The compounds of the present invention may exist in one crystal structure or in multiple crystal structures, i.e., have "polymorphism". The compounds of the present invention may exist in different crystal forms.

[0165] As used herein, the term "stereoisomer" includes conformational isomers and configurational isomers, wherein the configurational isomers mainly include cis-trans isomers and optical isomers. The compounds of the present invention can exist in the form of stereoisomers, and therefore encompass all possible stereoisomer forms, and any combination or any mixture thereof. For example, a single enantiomer, a single diastereomer or a mixture of the above. When the compounds of the present invention contain an olefin double bond, unless otherwise specified, it includes cis-isomers and trans-isomers, and any combination thereof.

[0166] As used herein, the term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom in a molecule between two positions, such as tautomers of the enol form and the keto form. If tautomers exist for the compounds of the present invention, they may exist in the form of a single tautomer or a mixture thereof, preferably in the form of a more stable tautomer as the main component.

[0167] As used herein, the term "prodrug" refers to a compound that can be converted into the present invention in the subject by reactions such as oxidation, reduction, hydrolysis, etc. The prodrug itself may or may not have the biological activity of the compound of formula (I) (e.g., regulating glucose and lipid metabolism activity, anti-inflammatory activity, antioxidant activity). For example, a compound of formula (I) including a hydroxyl group or a carboxyl group can be administered in the form of an ester, which is hydrolyzed and converted into a hydroxyl compound or a carboxyl compound in the body. Similarly, a compound of formula (I) including an amino group undergoes acylation, alkylation or phosphorylation to form, for example, eicosanoylamino, alanylamino, pivaloyloxymethylamino compounds for administration. Further information on the use of prodrugs can be found in Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T Higuchi and W Stella) and Bioreversible Carriers in Drug Design, Pergamon Press, 1987 (ed. EB Roche, American Pharmaceutical Association). Some examples of prodrugs according to the present invention include: (i) if the compound of formula (I) contains a carboxylic acid functional group (-COOH), then esters thereof are included, for example, using (C 1 -C 8 ) alkyl group instead of hydrogen; (ii) if the compound of formula (I) contains an alcohol function (-OH), then its ethers are included, for example, with (C 1 -C 6 ) alkanoyloxymethyl group instead of hydrogen; and (iii) if the compound of formula (I) contains a primary or secondary amino functional group (-NH 2 or -NHR, wherein R is not H), then includes its amides, for example, with (C 1 -C 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).

[0168] As used herein, the term "pharmaceutically acceptable carrier or excipient" refers to a carrier and / or excipient that is pharmacologically and / or physiologically compatible with a subject and an active ingredient, which is well known in the art (see, e.g., Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and includes, but is not limited to, disintegrants, binders, surfactants, glidants, lubricants, pH adjusters, ionic strength enhancers, agents that maintain osmotic pressure, agents that delay absorption, diluents, antioxidants, colorants, flavoring agents, preservatives, taste masking agents, etc. For example, non-limiting examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, cross-linked sodium carboxymethylcellulose, cross-linked polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropyl cellulose, starch, pregelatinized starch, and sodium alginate. Non-limiting examples of binders include microcrystalline cellulose, gelatin, sugar, polyethylene glycol, natural and synthetic gums, polyvinyl pyrrolidone, pregelatinized starch, hydroxypropyl cellulose and hydroxypropyl methylcellulose. Non-limiting examples of diluents include lactose (monohydrate, spray-dried monohydrate, anhydrous, etc.), mannitol, xylitol, glucose, sucrose, sorbitol, microcrystalline cellulose, starch and calcium hydrogen phosphate dihydrate. Non-limiting examples of surfactants include sodium lauryl sulfate and polysorbate 80. Non-limiting examples of glidants include silicon dioxide and talc. Non-limiting examples of lubricants include magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate and a mixture of magnesium stearate and sodium lauryl sulfate. Non-limiting examples of pH regulators include but are not limited to phosphate buffers. Ionic strength enhancers include but are not limited to sodium chloride. Reagents that maintain osmotic pressure include but are not limited to sugar, NaCl and the like. Reagents that delay absorption include but are not limited to monostearate and gelatin. Preservatives include, but are not limited to, various antibacterial and antifungal agents such as thimerosal, 2-phenoxyethanol, parabens, chlorobutanol, phenol, sorbic acid, and the like.

[0169] As used herein, the term "prevention" refers to a method implemented in order to prevent or delay the occurrence of a disease or illness or symptom in a subject. As used herein, the term "treatment" refers to a method implemented in order to obtain a beneficial or desired clinical result. For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, alleviating symptoms, reducing the scope of the disease, stabilizing (i.e., no longer worsening) the state of the disease, delaying or slowing the development of the disease, improving or alleviating the state of the disease, and alleviating symptoms (whether partially or completely), whether detectable or undetectable. In addition, "treatment" can also refer to, compared with the expected survival period (if not receiving treatment), extending the survival period.

[0170] As used herein, the term "effective amount" refers to an amount sufficient to obtain or at least partially obtain a desired effect. For example, a disease prevention (e.g., metabolic disease or the symptom associated with a metabolic disease) effective amount refers to an amount sufficient to prevent, prevent, or delay the occurrence of a disease (e.g., metabolic disease or the symptom associated with a metabolic disease); a disease treatment effective amount refers to an amount sufficient to cure or at least partially prevent the disease and its complications of a patient who has a disease. It is entirely within the capabilities of those skilled in the art to determine such an effective amount. For example, the effective amount for therapeutic use will depend on the severity of the disease to be treated, the overall state of the patient's own immune system, the patient's general condition such as age, weight and sex, the mode of administration of the drug, and other treatments administered simultaneously, etc. DETAILED DESCRIPTION

[0171] In the following examples, the structures of the compounds were determined by nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LCMS). NMR chemical shifts (δ) are expressed in 10 -6 The NMR measurements were performed using a Bruker AVANCE III 500 MHz NMR instrument and the solvent used was deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated chloroform (CDCl 3 ), deuterated methanol (CD 3 OD), etc., and tetramethylsilane (TMS) was the internal standard.

[0172] Analytical liquid chromatography: Analytical liquid chromatography is used to monitor the reaction system and analyze the purity of important intermediates and target products. Analytical liquid chromatography system: Agilent 1260 Infinity II (equipped with 1260 Binary Pump, Vial Sampler, MCT column oven, DAD WR detector); Chromatographic column: Waters XBridge BEH C18 3.5um, 4.6*150mm.

[0173] Preparative liquid chromatography uses preparative liquid phase to separate and purify a small amount of important intermediates and target products. Preparative liquid phase system: Agilent 1290Infinity II (equipped with 1290Prep Bin Pump, Prep ALS / FC injector / collector, 1260VWD detector); C18 reverse phase silica gel column.

[0174] Liquid chromatography-mass spectrometry uses LCMS for routine reaction system detection and molecular weight signal determination of target products. LC-MS system: Waters UPLC H-Class ultra-high performance liquid chromatography-QDa coupling instrument; chromatographic column: Waters XBridge BEH C181.7um, 2.1*75mm.

[0175] Liquid mobile phase ratio Acidic conditions: A: 0.05% formic acid aqueous solution, B: acetonitrile; Alkaline conditions: A: 0.05% ammonia aqueous solution, B: acetonitrile; Neutral conditions: A: 5mM NH 4 HCO 3 Aqueous solution, B: acetonitrile. The above data are the default mobile phase ratios, which can be appropriately adjusted according to the properties of the actual samples during the specific analysis and testing.

[0176] Rapid purification system Rapid purification system is used for rough preparation and purification of compounds in conventional reaction systems. The instrument model is Biotage Isolera One, and the normal phase column and reverse phase column used with it are purchased from Santai Tech. The common specifications of normal phase silica gel columns are 12g, 25g, 40g, 80g, 120g and 330g. The specific parameters are as follows: 40-63μm, The commonly used specifications of reverse phase C18 columns are 40g, 80g, 120g and 330g, and the specific parameters are as follows: 20-45μm,

[0177] 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.

[0178] Chemical reagents The known starting materials of the present invention can be synthesized by using or according to literature, patent methods, etc. known in the art, or can be purchased from chemical reagent companies such as Alfa Aesar, Acros, Sigma-Aldrich, TCI, Aicon, Anergy, Aladdin, J&K, Bid Pharmaceuticals, Leyan, McLean, Accela, Adamas, Xiens, and Inochem.

[0179] Abbreviations

[0180] Protecting Group Abbreviations:

[0181] Bn Benzyl

[0182] Boc tert-Butyloxycarbonyl

[0183] Cbz benzyloxycarbonyl

[0184] MOM Methoxymethyl

[0185] PMB p-Methoxybenzyl

[0186] TBDPS tert-Butyldiphenylsilyl

[0187] TBS tert-Butyldimethylsilyl

[0188] THP 2-Tetrahydropyranyl

[0189] Tos p-Toluenesulfonyl

[0190] Reagent Abbreviations:

[0191] ACN Acetonitrile

[0192] AIBN Azobisisobutyronitrile

[0193] Boc 2 O Di-tert-butyl dicarbonate

[0194] BPO Benzoyl Peroxide

[0195] tBuONa Sodium tert-butoxide

[0196] CAM Ammonium molybdate cerium sulfate color developer

[0197] CDI N,N'-Carbonyldiimidazole

[0198] m-CPBA m-Chloroperbenzoic acid

[0199] DABCO Triethylenediamine

[0200] DCE 1,2-Dichloroethane

[0201] DCM Dichloromethane

[0202] DEAD Diethyl azodicarboxylate

[0203] DIAD Diisopropyl azodicarboxylate

[0204] DIBAL-H Diisobutylaluminum hydride

[0205] DBU 1,8-diazobispiro[5.4.0]undec-7-ene

[0206] DHP 3,4-Dihydro-2H-pyran

[0207] DIEA N,N-Diisopropylethylamine

[0208] DMAP 4-dimethylaminopyridine

[0209] DMF N,N-Dimethylformamide

[0210] DMSO Dimethyl sulfoxide

[0211] EA Ethyl acetate

[0212] HMPA Hexamethylphosphoramide

[0213] HATU N,N,N′,N′-Tetramethyl-O-(7-azabenzotriazole-1-

[0214] Urea hexafluorophosphate

[0215] LAH Lithium Aluminum Hydride

[0216] NBS N-Bromosuccinimide

[0217] PE Petroleum Ether

[0218] PMA Phosphomolybdic Acid

[0219] PyBOP 1H-Benzotriazol-1-yloxytripyrrolidino hexafluorophosphate TBAF Tetrabutylammonium fluoride

[0220] TBDPSCl tert-Butyldiphenylsilyl chloride

[0221] TBSCl tert-Butyldimethylsilyl chloride

[0222] TEA Triethylamine

[0223] TFA Trifluoroacetic acid

[0224] THF Tetrahydrofuran

[0225] TPSCl 2,4,6-Triisopropylbenzenesulfonyl chloride

[0226] TsCl p-Toluenesulfonyl chloride

[0227] TsOH p-Toluenesulfonic acid

[0228] Example

[0229] Example 1: Synthesis of Compound A-1 / A-21

[0230]

[0231] Step A: At room temperature, dissolve 1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-3 4 -Methyl carboxylate (50 mg, 119.8 μmol) in tetrahydrofuran (1 mL) was added with lithium aluminum tetrahydride (2.5 M, 95.8 μL). After the addition, the whole mixture was stirred at 0°C for 0.5 hours. Ice water (1 mL) and 10% sodium hydroxide solution were added to the reaction solution, extracted with ethyl acetate (2 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. Then, it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (FA)-MeOH]; gradient: 20%-60% B in 36 minutes) to obtain compound A-21, a white solid product (1 4 -Amino-1 1 H-5,8-Dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzoylcyclooctane-3 4 -yl)methanol formate (260 μg, 0.6%). 1 H NMR (400 MHz, DMSO-d 6 )δppm 8.43(s,1H),8.39-8.47(m,1H),7.86-7.94(m,2H),7.53(brs,3H),7.30(d,J=7.9Hz,1H),7.03-7.15(m,2H),6.85(d,J=7.5Hz,1H),6.66(br d,J=8.0Hz,1H),5.36(br s,4H),5.34(s,2H),4.63(br d,J=3.9Hz,2H). ESI-MS m / z=390.1[M+H] + ; Calcu.=389.1. And compound A-1, white solid compound 1 1 H-5,8-Dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-14 -amine (2.83 mg, 7%). 1 H NMR (400 MHz, DMSO-d 6 )δppm7.98(s,1H),7.90(s,1H),7.49(br s,3H),7.26-7.32(m,1H),7.21(t,J=7.7Hz,1H),7.04-7.15(m,2H),6.85(d,J=7.8Hz,1H),6.65-6.74(m,1H),5.37(s,4H),5.24(s,2H). ESI-MSm / z=360.1[M+H] + ;Calcu.=359.1.

[0232] Example 2: Synthesis of Compound A-2

[0233]

[0234] Step A: At room temperature, add tert-butylchlorodimethylsilane (31.8 g, 211 mmol) and imidazole (21.6 g, 317 mmol) to tetrahydrofuran (100 mL) in which (3-aminophenyl)methanol (13.0 g, 101 mmol) is dissolved. After the addition, the whole mixture is stirred at 25°C for 16 hours. Add water (100 mL) to the reaction solution and extract it three times with ethyl acetate (100 mL*3). Dry the organic phase with magnesium sulfate, filter and concentrate. Use flash silica gel chromatography ( 330g Silica gel chromatography column, eluent 0-5% ethyl acetate / petroleum ether) to obtain a yellow oily product 3-((tert-butyl(dimethyl)silyl)oxymethyl)aniline (25.9 g, crude product). ESI-MS m / z=238.0[M+H] + ;Calcu.=237.2.

[0235] Step B: At room temperature, di-tert-butyl dicarbonate (12.1 g, 55.6 mmol) was added to ethanol (150 mL) in which 3-((tert-butyl(dimethyl)silyl)oxymethyl)aniline (12.0 g, 50.5 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 16 hours. The reaction solution was poured into water (150 mL), extracted with ethyl acetate (200 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated to obtain tert-butyl N-(3-((tert-butyl(dimethyl)silyl)oxymethyl)phenyl)carbamate (17.0 g, crude product) as a white solid. 1 H NMR: (400MHz, DMSO-d 6)δppm 9.25(s,1H),7.36(s,1H),7.25(br d,J=8.0Hz,1H),7.12(t,J=7.8Hz,1H),6.81(br d,J=7.5Hz,1H),4.57(s,2H),1.40(s,9H),0.83(s,9H),0.00(s,6H).

[0236] Step C: At 0°C, sodium hydride (1.42 g, 35.6 mmol, 60% purity) was added to N,N-dimethylformamide (100 mL) in which tert-butyl N-(3-((tert-butyl(dimethyl)silyl)oxymethyl)phenyl)carbamate (6.00 g, crude product) was dissolved, and the mixture was stirred at 25°C for 30 minutes. 1,3-bis(chloromethyl)benzene (15.6 g, 88.9 mmol) was then added to the mixture, and the mixture was stirred at 25°C for 1.5 hours. The reaction solution was poured into water (15 mL), extracted with ethyl acetate (20 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product, tert-butyl N-(3-((tert-butyl(dimethyl)silyl)oxymethyl)phenyl)-N-((3-(chloromethyl)phenyl)methyl)carbamate (7.70 g, crude product). 1 H NMR: (400MHz, DMSO-d 6 )δppm7.31-7.19(m,4H),7.17-7.10(m,2H),7.09-7.01(m,2H),4.79(s,2H),4.69(s,2H),4.62(s,2H),1.34(s,9H),0.84(s,9H),0.00(s,6H).

[0237] Step D: To N,N-dimethylformamide (140 mL) in which tert-butyl N-(3-((tert-butyl(dimethyl)silyl)oxymethyl)phenyl)-N-((3-(chloromethyl)phenyl)methyl)carbamate (7.60 g, 16.0 mmol) was dissolved was added 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2.71 g, 16.0 mmol) and potassium carbonate (4.41 g, 31.9 mmol) at room temperature and stirred at 25 °C for 16 hours. The mixture was diluted with water (50 mL), extracted with ethyl acetate (50 mL*3), and the organic layer was dried over magnesium sulfate, filtered and concentrated to give a yellow oily product N-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)methyl)-N-(3-((tert-butyl(dimethyl)silyl)oxymethyl)phenyl)-carbamic acid tert-butyl ester (9.00 g, crude product). ESI-MS m / z=609.3[M+H] + ;Calcu.=608.3.

[0238] Step E: At 0°C, tetrabutylammonium fluoride (1M, 22.2 mL) was added to tetrahydrofuran (150 mL) in which N-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)methyl)-N-(3-((tert-butyl(dimethyl)silyl)oxymethyl)phenyl)-carbamic acid tert-butyl ester (9.00 g, 14.8 mmol) was dissolved, and the mixture was stirred at 25°C for 2 hours. The mixture was diluted with water (100 mL), extracted with ethyl acetate (200 ml*3), and the organic layer was dried over magnesium sulfate, filtered and concentrated. The residue was purified by flash silica chromatography ( 80g Silica gel chromatography column, eluent 0-60% ethyl acetate / petroleum ether) was passed through the column to obtain yellow solid N-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)methyl)-N-(3-(hydroxymethyl)phenyl)carbamic acid tert-butyl ester (1.50 g, five-step yield 37%). ESI-MS m / z=495.2[M+H] + ;Calcu.=494.2.

[0239] Step F: At 0°C, sodium hydride (24.2 mg, 606 μmol, content 60%) was added to a solution of tert-butyl N-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)methyl)-N-(3-(hydroxymethyl)phenyl)carbamate (100 mg, 202 μmol) in N-methylpyrrolidone (1 mL), and stirred at 110°C for 2 hours. The mixture was filtered and then purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (FA)-MeOH]; gradient: 22%-62% B in 36 minutes) to obtain compound A-2, a white solid product 1 1 H-8-Oxa-5-aza-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-1 4 -amine (5.75 mg, 15.3 μmol, yield 8%). 1 H NMR: (400MHz, DMSO-d 6)δppm 7.90(s,1H),7.82(s,1H),7.33-7.74(m,2H),7.22-7.30(m,1H),7.14-7.22(m,1H),7.05(br d,J=7.5Hz,1H),6.95–7.01(m,1H),6.87(t,J=7.8Hz,1H),6.50(br d,J=7.5Hz,1H),6.28-6.43(m,2H),5.25-5.43(m,4H),4.19(br d,J=6.6Hz,2H). ESI-MS m / z=359.1[M+H] + ;Calcu.=358.2.

[0240] Example 3: Synthesis of Compound A-3

[0241]

[0242] Step A: Add a mixture of 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (17.84g, 24.39mmol) and triethylamine (49.35g, 487.8mmol, 67.9mL) to a solution of (3-bromo-5-fluoro-phenyl)methanol (50g, 243.9mmol) in methanol (500mL), degas with nitrogen for 3 times, and then stir at 100°C for 16 hours under a carbon monoxide atmosphere. The reaction mixture was diluted with water (500mL) and extracted with ethyl acetate (500mL*3). The organic phase was dried over magnesium sulfate, filtered and concentrated, and the residue was subjected to rapid silica gel chromatography ( 660g kuaisu Silica gel chromatography column, eluent 0-25% ethyl acetate / petroleum ether) to obtain black solid 3-fluoro-5-(hydroxymethyl)benzoic acid methyl ester (30g, 67%). ESI-MS m / z=185.2[M+H] + ,Calcu.=184.0.

[0243] Step B: Add triethylamine (26.37 g, 260.6 mmol, 36.28 mL) to a solution of methyl 3-fluoro-5-(hydroxymethyl)benzoate (16 g, 86.88 mmol) in dichloromethane (160 mL), add methanesulfonyl chloride (19.34 g, 168.8 mmol, 13.1 mL) at 0°C, and stir at 25°C for 2 hours. Pour the reaction solution into water (120 mL), extract with dichloromethane (150 mL*3), dry the organic phase with magnesium sulfate, filter and concentrate. Obtain a white solid product, methyl 3-fluoro-5-(methylsulfonyloxymethyl)benzoate (15.36 g, crude product).

[0244] Step C: To a solution of methyl 3-fluoro-5-(methylsulfonyloxymethyl)benzoate (10 g, crude product) in acetonitrile (100 mL), potassium carbonate (10.54 g, 76.26 mmol) and 3-(hydroxymethyl)phenol (4.78 g, 38.51 mmol) were added and stirred at 50°C for 16 hours. The mixture was filtered and concentrated. The product was purified by flash silica gel chromatography ( 40g fast The residue was purified by chromatography on a silica gel column with 0-27% ethyl acetate / petroleum ether as the eluent to obtain a pale yellow oily product, methyl 3-fluoro-5-((3-(hydroxymethyl)phenoxy)methyl)benzoate (7.37 g, two-step yield 45%). 1 H NMR (400 MHz, DMSO-d 6 )δ7.90(s,1H),7.54-7.70(m,2H),7.19-7.30(m,1H),6.99(br s,1H),6.84-6.94(m,2H),5.21(s,2H),5.19(brs,1H),4.47(br d, J=5.6Hz, 2H), 3.88 (d, J=1.0Hz, 3H). ESI-MS m / z=273.2[M-OH]+; Calcu.=290.1.

[0245] Step D: Add triethylamine (7.32 g, 72.34 mmol, 10.1 mL) to a solution of methyl 3-fluoro-5-((3-(hydroxymethyl)phenoxy)methyl)benzoate (7.0 g, 24.11 mmol) in dichloromethane (70 mL), add methanesulfonyl chloride (5.49 g, 47.93 mmol, 3.7 mL) at 0°C, and stir the mixture at 25°C for 4 hours. Pour the reaction solution into water (120 mL), extract with dichloromethane (150 mL*3), dry the organic phase with magnesium sulfate, filter and concentrate. Obtain a colorless oily product, methyl 3-fluoro-5-((3-(methylsulfonyloxymethyl)phenoxy)methyl)benzoate (7.1 g, crude product).

[0246] Step E: To a solution of methyl 3-fluoro-5-((3-(methylsulfonyloxymethyl)phenoxy)methyl)benzoate (3 g, 8.14 mmol) in N,N-dimethylformamide (70 mL), potassium carbonate (3.38 g, 24.43 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.38 g, 8.14 mmol) were added, followed by stirring at 50°C for 16 hours. The reaction solution was filtered and concentrated, and the residue was purified by flash silica gel chromatography ( 20g fast Silica gel chromatography column, eluent 0-35% tetrahydrofuran / petroleum ether) was passed through the column to obtain a light yellow solid product 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-5-fluoro-benzoic acid methyl ester (1.40 g, two-step yield 31%). ESI-MS m / z=442.1[M+H] + ;Calcu.=441.1.

[0247] Step F: At 0°C, diisobutylaluminum hydride (1M, 12.8 mL) was added to a solution of 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-5-fluoro-benzoic acid methyl ester (1.13 g, 2.56 mmol) in tetrahydrofuran (12 mL), and the mixture was stirred at 0°C for 2 hours. The mixture was quenched with water (10 ml) at 0°C, extracted with ethyl acetate (12 ml*3), the organic layer was dried over magnesium sulfate, filtered and concentrated to give a white solid product (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-5-fluoro-phenyl)methanol (0.84 g, 79%). ESI-MS m / z=414.1[M+H] + ;Calcu.=413.1.

[0248] Step G: Compound (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-5-fluoro-phenyl)methanol (100 mg, 241.6 μmol) was dissolved in N-methylpyrrolidone (1 mL) solution, sodium hydride (29.0 mg, 724.9 μmol, 60% purity) was added, and stirred at 110° C. for 3 hours. The mixture was filtered and concentrated, filtered and concentrated, and then purified by reverse phase high performance liquid chromatography (chromatographic column: C18 150*40mm*10um, mobile phase: [water (formic acid)-methanol], gradient: 37% to 74% B, 36 min) to obtain compound A-3, white solid product 6 5 -Fluoro-1 1 H-4,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-1 4 -amine (10 mg, 11%). 1 H NMR (400 MHz, DMSO-d 6)δ7.91-7.88(m,2H),7.78-7.45(m,2H),7.39(br s,1H),7.03-7.09(m,2H),6.94-7.00(m,1H),6.85-6.90(m,1H),6.71(dd,J=2.1,8.1Hz,1H),5.36(s,2H),5.27(s,2H),5.22-5.25(m,2H). ESI-MS m / z=378.1[M+H]+; Calcu.=377.1.

[0249] Example 4: Synthesis of Compound A-4

[0250]

[0251] Step A: At 0°C, add imidazole (288 mg, 4.23 mmol) to tetrahydrofuran (20 mL) in which (5-fluoro-1,3-phenylene) dimethanol (600 mg, 3.84 mmol) and TBDPSCl (1.16 g, 4.23 mmol) are dissolved. After the addition, the entire mixture is stirred at 0°C for 3 hours. The reaction solution is warmed to room temperature, and then water (50 mL) is added. It is extracted three times with ethyl acetate (30 mL*3). The organic phases are combined, washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a solid residue. The residue is separated by silica gel column (SiO 2 , PE:EA=50:50), the reaction gave a colorless oily product (3-((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorophenyl)methanol (800 mg, 77%). ESI-MS m / z=395.3[M+H] + ;Calcu.=394.2.

[0252] Step B: At 0°C and N 2 Under protection, diethyl azodicarboxylate (522 mg, 3 mmol) was added to tetrahydrofuran (20 mL) in which methyl 5-hydroxypyridine-3-carboxylate (153 mg, 1 mmol), (3-((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorophenyl)methanol (400 mg, 1.01 mmol), and triphenylphosphine (786 mg, 3 mmol) were dissolved. After the addition was completed, the whole mixture was stirred at 0°C overnight. The reaction solution was concentrated under reduced pressure to obtain a solid residue. The residue was separated by silica gel column (SiO 2 , PE:EA=50:50). The reaction gave the product 5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxynicotinic acid methyl ester (298 mg, 73%) as a light yellow oil. ESI-MS m / z=530.2[M+H]+ ;Calcu.=529.2.

[0253] Step C: At 0°C, lithium aluminum hydride (19 mg, 0.5 mmol) was added to tetrahydrofuran (20 mL) in which methyl 5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxynicotinate (265 mg, 0.5 mmol) was dissolved. After the addition, the whole mixture was stirred at 0°C for 1 hour. After the reaction was complete, methanol (10 mL) was added under ice bath until no bubbles were generated, and the mixture was concentrated under reduced pressure to obtain a solid residue. The residue was separated by preparative TLC (SiO 2 , DCM:MeOH=95:5). The reaction gave a colorless oily product (5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxypyridin-3-yl)methanol (150 mg, 59%). ESI-MS m / z=502.2[M+H] + ;Calcu.=501.2.

[0254] Step D: At 0°C, N-bromosuccinimide (40 mg, 0.22 mmol) was added in batches to tetrahydrofuran (15 mL) in which (5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxypyridin-3-yl)methanol (70 mg, 0.14 mmol) and triphenylphosphine (59 mg, 0.22 mmol) were dissolved. After the addition, the whole mixture was stirred at 0°C for 2 hours. The reaction solution was concentrated under reduced pressure to obtain a yellow crude product 3-(bromomethyl)-5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxy)pyridine, which was used directly in the next step without purification.

[0255] Step E: At room temperature, cesium carbonate (34.6 mg, 0.106 mmol) was added to acetonitrile (10 mL) in which 3-(bromomethyl)-5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxy)pyridine (40 mg, 0.071 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (12 mg, 0.071 mmol) were dissolved. After the addition was complete, the entire mixture was stirred at room temperature for 3 hours. After the reaction was complete, the reaction solution was directly concentrated to obtain a solid residue. The residue was subjected to HPLC preparation and separation. Preparation conditions: Waters XBridge C18 (19 mm×250 mm, 5 μm); A: 0.1% formic acid in water; B: MeCN; 18 mL / min; 0~3 min, 15% B; 3.5~15 min, 30%~100% B; detector, UV 254 nm. The reaction gave a white solid product, 1-((5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxypyridin-3-yl))methyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (13 mg, two-step yield 28%). ESI-MS m / z=653.1[M+H] + ;Calcu.=652.2.

[0256] Step F: At room temperature, cesium carbonate (65 mg, 0.2 mmol) and lithium iodide (27 mg, 0.2 mmol) were added to acetonitrile (5 mL) in which 1-((5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxypyridin-3-yl))methyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (13 mg, 0.02 mmol) was dissolved. After the addition, the whole mixture was controlled or heated to 110° C. and stirred for 18 hours. After the reaction was complete, the reaction solution was directly concentrated to obtain a solid residue. The residue was subjected to HPLC preparation and separation, preparation conditions: Waters XBridge C18 (19mm×250mm, 5μm); A: 0.1% formic acid in water; B: MeCN; 18mL / min; 0~3min, 5% B; 3.5~15min, 10%~100% B; detector, UV 254nm. The reaction gave compound A-4, white solid product 6 5 -Fluoro-1 1 H-4,8-dioxa-1-(1,6)-pyrazolo[3,4-d]pyrimidine-3(3,5)-pyridine-6(1,3)-benzocyclooctylamine-1 4 -amine (0.75 mg, 10%). 1H NMR (500MHz, CD 3 OD): δppm8.19(d,J=3Hz,1H),8.10(s,2H),7.42-7.46(m,1H),7.37-7.41(m,1H),6.96-6.97 (d, J=8Hz, 1H), 6.85-6.87 (d, J=8Hz, 1H) 5.44-5.48 (m, 2H), 5.35 (t, J=5Hz, 2H), 5.21 (s, 2H). ESI-MS m / z=378.9[M+H] + ;Calcu.=378.1.

[0257] Example 5: Synthesis of Compound A-5 / M-1

[0258]

[0259] Step A: In dichloromethane (150 mL) dissolved with (4-bromo-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (9 g, 20.57 mmol), triethylamine (4.16 g, 41.15 mmol, 5.73 mL) and acetyl chloride (1.94 g, 24.69 mmol, 1.76 mL) were added. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into ice water (150 mL), extracted with ethyl acetate (150 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. A yellow oily product (4-bromo-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)acetate (9.8 g, crude product) was obtained.

[0260] Step B: Tetrabutylammonium fluoride (1M, 30.7 mL) was added to tetrahydrofuran (150 mL) in which (4-bromo-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)acetate (9.8 g, 20.44 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into ice water (150 mL), extracted with ethyl acetate (150 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated by silica gel column (silica, 0-33%, petroleum ether:ethyl acetate, @60 mL / min) to obtain a yellow oily product (4-bromo-3-((3-(hydroxymethyl)phenoxy)methyl)phenyl)acetate (3.2 g, two-step yield 43%). 1 H NMR (400 MHz, DMSO-d 6)δ7.69(d,J=8.1Hz,1H),7.60(d,J=1.9Hz,1H),7.31(dd,J=2.0,8.1Hz,1H),7.23-7.29(m,1H),7.01(s ,1H),6.87-6.95(m,2H),5.20(t,J=5.8Hz,1H),5.07-5.12(m,4H),4.49(d,J=5.8Hz,2H),2.06(s,3H).

[0261] Step C: At 0°C, add methanesulfonyl chloride (2.18 g, 19.03 mmol, 1.5 mL) and triethylamine (2.66 g, 26.29 mmol, 3.7 mL) to dichloromethane (50 mL) in which (4-bromo-3-((3-(hydroxymethyl)phenoxy)methyl)phenyl)acetate (3.2 g, 8.76 mmol) is dissolved. After the addition, the whole mixture is stirred at 25°C for 2 hours. The reaction solution is poured into ice water (50 mL), extracted with ethyl acetate (50 mL*3), the organic phase is dried over magnesium sulfate, filtered and concentrated. A yellow oily product (4-bromo-3-((3-(methylsulfonyloxymethyl)phenoxy)methyl)phenyl)acetate (3.8 g, crude product) is obtained.

[0262] Step D: Potassium carbonate (2.36 g, 17.10 mmol) and 6-chloro-2H-pyrazolo[3,4-d]pyrimidin-4-amine (1.45 g, 8.55 mmol) were added to N,N-dimethylformamide (50 mL) in which methyl (4-bromo-3-((3-(methylsulfonyloxymethyl)phenoxy)methyl)phenyl)acetate (3.79 g, 8.55 mmol) was dissolved. After the addition, the whole mixture was stirred at 40°C for 16 hours, the reaction solution was poured into water (50 mL), extracted with ethyl acetate (50 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was purified by silica gel column separation (silica, 0-35% petroleum ether: ethyl acetate, @80 mL / min) to obtain a yellow solid product (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-bromo-phenyl)acetate (1.7 g, two-step yield 38%). ESI-MS m / z=518.0[M+H] + ;Calcu.=517.0.

[0263] Step E: At 0°C, sodium hydroxide (263 mg, 6.58 mmol) and water (10 mL) were added to tetrahydrofuran (10 mL) in which (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-bromo-phenyl)acetic acid methyl ester (1.7 g, 3.29 mmol) was dissolved. After the addition was complete, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into water (10 mL), extracted with ethyl acetate (10 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated by silica gel column (silica, 0-49% petroleum ether: ethyl acetate, @80 mL / min) to obtain a white solid product (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-bromo-phenyl)methanol (1.55 g, 99%). ESI-MS m / z=476.0[M+H] + ;Calcu.=475.0.

[0264] Step F: At room temperature, sodium hydride (379 mg, 9.48 mmol, 60% purity) was added to N-methylpyrrolidone (15 mL) in which (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-bromo-phenyl)methanol (1.5 g, 3.16 mmol) was dissolved. After the addition, the whole mixture was stirred at 110°C for 2 hours. The reaction solution was poured into water (15 mL), extracted with ethyl acetate (15 mL*3), the organic phase was washed with brine and dried over magnesium sulfate, filtered and concentrated. M-1, a white solid product 6, was obtained. 6 -Bromo-1 1 H-4,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (400 mg, crude).

[0265] Step G: At 0°C, dissolve 6 6 -Bromo-1 1 H-4,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4-amine (50mg, 114.08μmol) in 1,4-dioxane (0.6mL), sodium carbonate (36.3mg, 342.2μmol), water (0.2mL), methylboric acid (34.2mg, 0.57mmol) and 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (8.4mg, 11.41μmol) were added. After the addition, the whole mixture was stirred at 80°C for 16 hours. The reaction solution was poured into water (1mL), extracted with ethyl acetate (1mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. Then, it was purified by reverse phase high performance liquid chromatography (chromatographic column: chromatographic column: C18 150*40mm*10um, mobile phase: [water (formic acid)-methanol], gradient: 36%-76% Bover 36min) to obtain compound A-5, white solid product 6 6 -methyl-1 1 H-4,8-Dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzoylcyclooctane-1 4 -amine (14.7 mg, 34%). 1 H NMR (400 MHz, DMSO-d 6 )δ7.96(s,1H),7.88(s,1H),7.51(br s,3H),7.15(d,J=8.3Hz,1H),7.01-7.09(m,2H),6.87(d,J=7.6Hz,1H),6.66-6.71(m,1H),5.31(br d,J=3.5Hz,4H),5.29(s,2H),2.31(s,3H). ESI-MS m / z=374.1[M+H] + ;Calcu.=373.2.

[0266] Example 6: Synthesis of Compound A-6

[0267]

[0268] Step A: Add potassium carbonate (28.77 g, 208.2 mmol) and iodomethane (23.64 g, 166.5 mmol, 10.4 mL) to N,N-dimethylformamide (250 mL) in which methyl 3-formyl-4-hydroxybenzoate is dissolved and stirred at 25°C for 12 hours. Pour the reaction into water (500 mL) and ethyl acetate (500 mL*2) and separate the layers. The organic layer is washed with brine (500 mL*2) and the aqueous layer is extracted with ether. The organic layers are combined, dried over magnesium sulfate, filtered, and concentrated in vacuo to give compound 3-formyl-4-methoxy-benzoic acid methyl ester (26 g, 96%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6 )δ10.33(s,1H),8.09-8.31(m,2H),7.36(d,J=8.8Hz,1H),4.00(s,3H),3.85(s,3H).

[0269] Step B: Sodium borohydride (10.21 g, 269.87 mmol) was added in batches to a methanol solution (250 mL) of 3-formyl-4-methoxy-benzoic acid methyl ester (26 g, 133.89 mmol) at 0°C and stirred at 25°C for 2 hours. The mixture was concentrated to remove methanol, diluted with water (120 mL) at 0°C, extracted with ethyl acetate (120 mL*3), and dried over magnesium sulfate. The mixture was filtered and concentrated under reduced pressure to obtain compound 3-(hydroxymethyl)-4-methoxy-benzoic acid methyl ester (26 g, 99%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ7.96–8.06(m,1H),7.87(dd,J=2.2,8.6Hz,1H),7.06(d,J=8.6Hz,1H),5.20(t,J=5.7Hz,1H),4.50(d,J=5.8Hz,2H),3.85(s,3H),3.82(s,3H).

[0270] Step C: Dichloromethane (150 mL) was added to 3-(hydroxymethyl)-4-methoxy-benzoic acid methyl ester (15 g, 76.45 mmol), followed by triethylamine (23.21 g, 229.4 mmol, 31.9 mL) and methanesulfonyl chloride (22.01 g, 192.2 mmol, 14.9 mL) and stirred at 0°C for 2 hours. The reaction mixture was quenched with water (100 mL) at 0°C. The reaction mixture was extracted with dichloromethane (100 mL*3). Drying over magnesium sulfate, filtering and concentrating under reduced pressure gave compound 4-methoxy-3-(methylsulfonyloxymethyl)benzoic acid methyl ester (16.67 g, 79%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d 6 )δ8.00-8.04(m,1H),7.97(dd,J=2.3,8.6Hz,1H),7.14-7.21(m,1H),4.76(s,2H),3.92(s,3H),3.81-3.84(m,3H).

[0271] Step D: Add 3-(chloromethyl)-4-methoxy-benzoic acid methyl ester (16.17 g, 75.33 mmol), cesium carbonate (73.64 g, 226.0 mmol) and 3-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (17.96 g, 75.33 mmol) to acetonitrile (160 mL) and stir at 50 ° C for 12 hours. Pour the reaction into water (500 mL) and ethyl acetate (500 mL*2) and separate the layers. The organic layer is washed with 500 mL*2 brine, the aqueous layers are combined, and extracted with ether. The combined organic layers are dried over magnesium sulfate, filtered, and concentrated in vacuo, and the residue is purified by flash silica gel chromatography ( 80g fast Silica gel column, eluent 0-15% ethyl acetate / petroleum ether gradient @100 mL / min) to give compound 3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-4-methoxy-benzoic acid methyl ester (22 g, 70%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ7.80–8.01(m,2H),7.25(t,J=7.9Hz,1H),7.16(d,J=8.6Hz,1H),6.92(s,1H),6.88(br d,J=8.0Hz,2H),5.08(s,2H),4.66(s,2H),3.90(s,3H),3.79(s,3H),0.87(s,9H),0.04(s,6H).

[0272] Step E: Tetrabutylammonium fluoride (1M, 77.42 mL) was added to a solution of 3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-4-methoxy-benzoic acid methyl ester (21.5 g, 51.61 mmol) in tetrahydrofuran (200 mL). Stir at 25 °C for 2 hours. The reaction mixture was quenched with water (100 mL) at 0 °C. The reaction mixture was extracted with ethyl acetate (100 mL*3). Dry over magnesium sulfate, filter and concentrate under reduced pressure to obtain a residue. Flash silica gel chromatography ( 80g fast Silica gel column, eluent: 0-20% tetrahydrofuran / petroleum ether gradient @100 mL / min) to give yellow oily compound 3-((3-(hydroxymethyl)phenoxy)methyl)-4-methoxy-benzoic acid methyl ester (16 g, crude product). 1 HNMR (400 MHz, DMSO-d 6)δ8.02(d,J=2.0Hz,1H),7.97(dd,J=2.1,8.6Hz,1H),7.16-7.29(m,2H),6.98(s,1H),6.82-6. 93(m,2H),5.19(t,J=5.8Hz,1H),5.09(s,2H),4.48(d,J=5.8Hz,2H),3.93(s,3H),3.82(s,3H).

[0273] Step F: Methanesulfonyl chloride (18.45 g, 161.1 mmol, 12.5 mL) and triethylamine (13.39 g, 132.3 mmol, 18.4 mL) were added to a solution of 3-((3-(hydroxymethyl)phenoxy)methyl)-4-methoxy-benzoic acid methyl ester (16 g, crude) in dichloromethane (160 mL) and stirred at 0°C for 2 hours. The reaction mixture was quenched with water (100 mL) at 0°C. The reaction mixture was extracted with dichloromethane (100 mL*3). Dry over magnesium sulfate, filter and concentrate under reduced pressure to give compound 4-methoxy-3-((3-(methylsulfonyloxymethyl)phenoxy)methyl)benzoic acid methyl ester (22.8 g, crude) as a white solid. 1 HNMR (400 MHz, DMSO-d 6 )δ8.01(s,1H),7.96(br dd,J=1.9,8.6Hz,1H),7.26-7.37(m,1H),7.17(br d,J=8.8Hz,1H),7.11(br d,J=2.3Hz,1H),7.00-7.06(m,1H),6.97(br dd,J=2.4,8.3Hz,1H),5.06-5.14(m,2H),4.72(s,1H),3.91(s,3H),3.81(s,3H),2.52(d,J=0.8Hz,3H).

[0274] Step G: Add potassium carbonate (2.18 g, 15.77 mmol), methyl 4-methoxy-3-((3-(methylsulfonyloxymethyl)phenoxy)methyl)benzoate (2 g, 5.26 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (891.5 mg, 5.26 mmol) to N,N-dimethylformamide (20 mL). Stir at 25°C for 12 hours. Filter the mixture, concentrate the mother liquor and flash silica gel chromatography ( 40g fast Silica gel column, eluent 0-40% tetrahydrofuran / petroleum ether gradient @100 mL / min) to obtain compound 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-methoxybenzoic acid methyl ester (450 mg, three-step yield 22%) as a white solid. ESI-MS m / z=454.1[M+H] + ;Calcu.=453.1.

[0275] Step H: 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-methoxy-benzoic acid methyl ester (300 mg, 661.0 μmol) was added to tetrahydrofuran (10 mL), and diisobutylaluminum hydride (1 M, 3.3 mL) was added dropwise at 0°C. The mixture was stirred at 25°C for 2 hours. The mixture was heated to 40°C with H 2 O (50mL), extracted with ethyl acetate (50mL*3), and dried over magnesium sulfate. Filtered and concentrated under reduced pressure to obtain the compound (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-methoxy-phenyl)methanol (450mg, crude product) as a white solid. ESI-MS m / z=408.2[M-OH] + ;Calcu.=425.1.

[0276] Step I: At 0°C, sodium hydride (28.2 mg, 704.4 μmol, 60% purity) was added to a solution of (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-methoxy-phenyl)methanol (100 mg, 234.8 μmol) in N-methylpyrrolidone (2 mL). Stir at 110°C for 12 hours. The reaction mixture was quenched with water (2 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. Purification by high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um, mobile phase: [water (ammonium bicarbonate)-acetonitrile], gradient: 16%-56% B over 1 min) to obtain compound A-6, 6 6 -Methoxy-1 1 H-4,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (4.87 mg, 9% yield in two steps) as a white solid. 1 HNMR (400 MHz, DMSO-d 6)δ8.04(d,J=1.8Hz,1H),7.88(s,1H),7.67-7.28(m,4H),7.10-7.02(m,1H),6.91(d,J=8.5Hz,1H),6. 87(d,J=7.4Hz,1H),6.67(dd,J=2.3,7.8Hz,1H),5.32(s,2H),5.29(s,2H),5.21(s,2H),3.74(s,3H). ESI-MS m / z=390.2[M+H] + ;Calcu.=389.2.

[0277] Example 7: Synthesis of Compound A-7

[0278]

[0279] Step A: At room temperature, concentrated sulfuric acid (530 mg, 5.40 mmol, 288 μL) was added to methanol (50 mL) in which 4-bromo-3-(hydroxymethyl)benzoic acid (4.8 g, 20.78 mmol) was dissolved. After the addition, the whole mixture was stirred at 80°C for 16 hours. The reaction mixture was evaporated, and a saturated sodium bicarbonate solution (150 mL) was slowly added. The mixture was extracted with ethyl acetate (150 mL*3). The organic phase was dried over magnesium sulfate, filtered and concentrated. A yellow solid product, methyl 4-bromo-3-(hydroxymethyl)benzoate (4.98 g, 98%), was obtained. 1 HNMR (400 MHz, DMSO-d 6 )δppm 8.09-8.16(m,1H), 7.68-7.79(m,2H), 5.64(t,J=5.6Hz,1H), 4.55(d,J=5.6Hz,2H), 3.87(s,3H)

[0280] Step B: At 0°C, add thionyl chloride (23.79 g, 199.94 mmol, 14.52 mL) to carbon tetrachloride (50 mL) in which methyl 4-bromo-3-(hydroxymethyl)benzoate (4.9 g, 19.99 mmol) is dissolved. After the addition, the whole mixture is stirred at 80°C for 16 hours. Pour the reaction solution into ice water (50 mL), extract with ethyl acetate (50 mL*3), dry the organic phase with magnesium sulfate, filter and concentrate. The residue is separated by silica gel column (silicon dioxide, 0-10%, petroleum ether: ethyl acetate, @100 mL / min) to obtain a white solid product methyl 4-bromo-3-(chloromethyl)benzoate (17 g, 66%). 1 H NMR (400 MHz, DMSO-d 6)δppm8.13(s,1H),7.78-7.76(m,2H),4.84(s,2H),3.81(s,3H)

[0281] Step C: At room temperature, 3-(((tert-butyldimethylsilyl)oxy)methyl)phenol (3.17 g, 13.28 mmol) and cesium carbonate (12.98 g, 39.85 mmol) were added to acetonitrile (40 mL) in which methyl 4-bromo-3-(chloromethyl)benzoate (3.5 g, 13.28 mmol) was dissolved. After the addition, the whole mixture was stirred at 50 °C for 16 hours. The reaction mixture was evaporated, saturated sodium bicarbonate solution (250 ml) was slowly added, and the mixture was extracted with ethyl acetate (150 ml*3). The organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated by silica gel column (silica, 0-4%, petroleum ether: ethyl acetate, @100 mL / min) to obtain a yellow oily product 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester (3.2 g, 52%). 1 H NMR (400 MHz, DMSO-d 6 )δppm8.05(s,1H),7.78-7.81(m,2H),7.23(t,J=7.8Hz,1H),6.82-6.93( m,3H),5.14(s,2H),4.64(s,2H),3.80(s,3H),0.83(s,9H),0.00(s,6H).

[0282] Step D: At 0°C, diisobutylaluminum hydride (1M, 32.23 mL) was added to tetrahydrofuran (30 mL) in which 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester (3 g, 6.45 mmol) was dissolved. After the addition, the whole mixture was stirred at 0°C for 2 hours. The reaction solution was poured into ice water (30 mL), extracted with ethyl acetate (30 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. A white solid product (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)phenyl)methanol (2.78 g, 99%) was obtained. 1 H NMR (400 MHz, DMSO-d 6 )δppm 7.58-7.44(m,2H),7.20(q,J=8.0Hz,2H),6.91-6.82(m,3H),5.24(t,J=5.7Hz, 1H), 5.03 (s, 2H), 4.63 (s, 2H), 4.41 (d, J = 5.6Hz, 2H), 0.83 (s, 9H), 0.00 (s, 6H).

[0283] Step E: At 0°C, methanesulfonyl chloride (1.32 g, 11.52 mmol, 0.89 mL) and triethylamine (1.04 g, 10.29 mmol, 1.43 mL) were added to dichloromethane (20 mL) in which (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)phenyl)methanol (1.5 g, 3.43 mmol) was dissolved. After the addition, the whole mixture was stirred at 0°C for 2 hours. The reaction solution was poured into ice water (5 mL), extracted with dichloromethane (5 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. A white solid product (3-((2-bromo-5-(chloromethyl)benzyl)oxy)phenyl)methanol (600 mg, 51%) was obtained. 1 H NMR (400 MHz, DMSO-d 6 )δppm 7.74-7.64(m,1H),7.39(dd,J=2.2,8.2Hz,1H),7.20-7.32(m,1H),7.01(s,1H),6.86 -6.97(m,1H),5.22(t,J=5.8Hz,1H),5.10(s,2H),4.79(s,2H),4.49(d,J=5.8Hz,2H).

[0284] Step F: At room temperature, 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (298 mg, 1.76 mmol) and potassium carbonate (485 mg, 3.51 mmol) were added to N,N-dimethylformamide (10 mL) in which (3-((2-bromo-5-(chloromethyl)benzyl)oxy)phenyl)methanol (600 mg, 1.76 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 16 hours. The reaction solution was poured into water (10 mL), extracted with ethyl acetate (10 mL*3), the organic phase was washed with saturated brine (10 mL), dried over magnesium sulfate, filtered and concentrated. A yellow solid product (3-((5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromobenzyl)oxy)phenyl)methanol (260 mg, 31%) was obtained. 1 H NMR (400 MHz, DMSO-d 6)δppm 8.16-8.39(m,2H), 7.64(d,J=8.1Hz,1H), 7.44(s,1H), 7.20-7.27(m,1H), 7.12(br d,J=6.5Hz,1H), 6.95(s,1H), 6.89-6.94(m,1H), 6.86-6.89(m,1H), 6.79-6.86 (m,1H), 5.45(s,2H), 5.19(t,J=5.8Hz,1H), 5.06(s,2H), 4.47(d,J=5.8Hz,2H).

[0285] Step G: At room temperature, sodium hydride (25.3 mg, 631.9 μmol, 60% purity) was added to N-methylpyrrolidone (1 mL) in which (3-((5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromobenzyl)oxy)phenyl)methanol (100 mg, 211 μmol) was dissolved. After the addition, the whole mixture was stirred at 110°C for 2 hours. The reaction solution was poured into ice water (1 mL), extracted with ethyl acetate (1 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated by silica gel column (silicon dioxide, 0-60%, petroleum ether:ethyl acetate, @40 mL / min) to obtain a white solid product 3. 4 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (25 mg, 27%). ESI-MS m / z = 440.0 [M+H] + ;Calcu.=439.0

[0286] Step H: At room temperature, dissolve 3 4 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (20 mg, 45.63 μmol) in tetrahydrofuran (0.4 mL), water (0.2 mL), potassium trifluoro((pyrrolidin-1-yl)methyl)borate (13.1 mg, 68.45 μmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (2.2 mg, 4.56 μmol), tris(dibenzylideneacetone)dipalladium (2.1 mg, 2.28 μmol) and sodium carbonate (14.5 mg, 136.9 μmol) were added. After the addition was complete, the flask was evacuated and N 2The mixture was ventilated 3 times and stirred at 80°C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. It was then purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (FA)-MeOH]; gradient: 6%-46% B in 36 minutes) to obtain compound A-7, a white solid product 3 4 -(pyrrolidin-1-ylmethyl)-1 1 H-5,8-Dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzoylcyclooctane-1 4 -amine formate (5.81 mg, 26%). 1 H NMR (400 MHz, DMSO-d 6 )δppm 8.25(s,1H), 7.91(s,1H), 7.89(s,1H), 7.59-7.75(m,1H), 7.54(s,1H), 7.36-7.51(m,1H), 7.21(d,J=7.8Hz,1H), 7.07(dd,J=15.8,8.00Hz ,2H), 6.81-6.87(m,1H), 6.67(dd,J=7.8,1.9Hz,1H), 5.42(s,2H), 5.35-5.38(m,2H), 5.34(s,2H), 3.66(s,2H), 2.40-2.42(m,2H), 1.69(br s,4H),1.24(br s,2H). ESI-MS m / z=443.2[M+H] + ;Calcu.=442.2.

[0287] Example 8: Synthesis of Compound A-8 / M-2

[0288]

[0289] Step A: Add imidazole (17.10 g, 251.19 mmol) and tert-butylchlorodimethylsilane (12.62 g, 83.73 mmol, 10.30 mL) to tetrahydrofuran (350 mL) in which 2-bromo-5-(hydroxymethyl)phenol (17 g, 83.73 mmol) is dissolved. After the addition, the whole mixture is stirred at 25 ° C for 16 hours. The reaction solution is poured into ice water (350 mL), extracted with ethyl acetate (350 mL*3), and the organic phase is dried with magnesium sulfate, filtered and concentrated. The residue is separated by silica gel column (silica, 0-5%, petroleum ether: ethyl acetate, @100 mL / min) to obtain a yellow oily product 2-bromo-5-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (19 g, 72%). 1H NMR (400 MHz, DMSO-d 6 )δ10.09(s,1H),7.32(d,J=8.1Hz,1H),6.80-6.93(m,1H),6.50-6.66(m,1H),4.53(s,2H),0.83(s,10H),0.00(s,6H).

[0290] Step B: Add cesium carbonate (36.97 g, 113.46 mmol) and 2-bromo-5-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (12 g, 37.82 mmol) to acetonitrile (250 mL) in which 3-(bromomethyl)benzaldehyde (9.03 g, 45.38 mmol) was dissolved. After the addition, the whole mixture was stirred at 50 °C for 16 hours. The reaction solution was poured into ice water (250 mL), extracted with ethyl acetate (250 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. A white oily product 3-((2-bromo-5-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzaldehyde (15.1 g, 92%) was obtained. 1 H NMR (400 MHz, DMSO-d 6 )δ10.00(s,1H),7.96(s,1H),7.85(d,J=7.6Hz,1H),7.76(d,J=7.8Hz,1H),7.58-7.66(m,1H),7. 05-7.10(m,1H),6.83(dd,J=1.6,8.1Hz,1H),5.28(s,2H),4.62(s,2H),0.83(s,9H),0.00(s,6H).

[0291] Step C: Add tetrabutylammonium fluoride (1M, 39.96 mL) to tetrahydrofuran (150 mL) in which 3-((2-bromo-5-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzaldehyde (11.6 g, 26.64 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into ice water (150 mL), extracted with ethyl acetate (150 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. The yellow oily product 3-((2-bromo-5-(hydroxymethyl)phenoxy)methyl)benzaldehyde (12 g, crude product) was obtained.

[0292] Step D: Add methanesulfonyl chloride (1.960 g, 17.11 mmol, 1.32 mL) and triethylamine (2.36 g, 23.35 mmol, 3.25 mL) to dichloromethane (50 mL) at 0°C. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into water (50 mL), extracted with dichloromethane (50 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. A white solid product (4-bromo-3-((3-formylphenyl)methoxy)phenyl)methanesulfonic acid methyl ester (3.1 g, 100%) was obtained. 1 H NMR (400 MHz, DMSO-d 6 )δ10.04(s,1H),8.03(s,1H),7.91(br d,J=7.4Hz,1H),7.83(br d,J=7.4Hz,1H),7.64-7.70(m,2H),7.34(s,1H),7.02(br d,J=8.0Hz,1H),5.34(s,2H),5.23(s,2H),3.22(s,3H).

[0293] Step E: To N,N-dimethylformamide (50 mL) in which methyl (4-bromo-3-((3-formylphenyl)methoxy)phenyl)methanesulfonate (3.1 g, 7.76 mmol) was dissolved, potassium carbonate (2.15 g, 15.53 mmol) and 6-chloro-2H-pyrazolo[3,4-d]pyrimidin-4-amine (1.32 g, 7.76 mmol) were added. After the addition, the whole mixture was stirred at 25°C for 16 hours. The reaction solution was poured into water (50 mL), extracted with ethyl acetate (10 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated on a silica gel column (silica, 0-35% petroleum ether:ethyl acetate, @80 mL / min) to give a yellow solid product 3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenoxy)methyl)benzaldehyde (1.3 g, yield 35%). 1 H NMR (400 MHz, DMSO-d 6 )δ10.03(s,1H),8.17-8.38(m,2H),8.11(s,1H),7.96(s,1H),7.87(s,1H),7.74(s,1H),7.64(br d,J=7.3Hz,1H),7.55(d,J=8.1Hz,1H),7.10(s,1H),6.68(br d,J=8.4Hz,1H),5.41(s,2H),5.27(s,2H).

[0294] Step F: At room temperature, sodium borohydride (120 mg, 3.17 mmol) and tetrahydrofuran (400 mL) were added to methanol (100 mL) in which 3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenoxy)methyl)benzaldehyde (1.3 g, 2.75 mmol) was dissolved. After the addition, the whole mixture was stirred at 0°C for 2 hours. The reaction solution was poured into water (400 mL), extracted with ethyl acetate (400 mL*3), the organic phase was washed with brine and dried over magnesium sulfate, filtered and concentrated. A yellow oily product (3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenoxy)methyl)phenyl)methanol (1.3 g, 2.74 mmol, yield 100%) was obtained. ESI-MS m / z=475.9[M+H] + ;Calcu.=475.0.

[0295] Step G: Sodium hydride (328.6 mg, 8.22 mmol, purity 60%) was added to N-methylpyrrolidone (10 mL) in which (3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenoxy)methyl)phenyl)methanol (1.3 g, 2.74 mmol) was dissolved. After the addition, the whole mixture was stirred at 110°C for 2 hours. The reaction solution was poured into water (10 mL), extracted with ethyl acetate (10 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated by silica gel column (silica, 0-49% petroleum ether: ethyl acetate, @80 mL / min) to obtain M-2, a white solid product 3. 4 -Bromo-1 1 H-4,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (400 mg, 33%). 1 HNMR (400 MHz, DMSO-d 6 )δ8.07(s,1H),7.90(s,1H),7.56(m,3H),7.35(br d,J=8.3Hz,1H),7.23-7.29(m,3H),6.78(br d,J=8.3Hz,1H),5.36(s,2H),5.34(s,2H),5.30(s,2H).

[0296] Step H: Dissolve 3 4 -Bromo-1 1 H-4,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-14 -amine (50mg, 114.1μmol) in tetrahydrofuran (0.5mL), potassium; trifluoro(pyrrolidin-1-ylmethyl) boride (21.8mg, 114.1μmol), water (0.5mL), sodium carbonate (36.3mg, 342.25μmol), (1E, 4E)-1,5-diphenylpenta-1,4-diene-3-one; palladium (10.45mg, 11.41μmol) and 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (5.44mg, 11.41μmol) were added. After the addition, the whole mixture was stirred at 80℃ for 16h. The reaction solution was poured into water (1mL), extracted with ethyl acetate (1mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. Then, the product was purified by reverse phase high performance liquid chromatography (chromatographic column: C18 150*40mm*10um, mobile phase: [water (formic acid)-methanol], gradient: 6%-46% B over 36min) to obtain compound A-8, a white solid product 3 4 -(pyrrolidin-1-ylmethyl)-1 1 H-4,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (2.6 mg, 5%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.30(br s,1H),8.03(s,1H),7.88(s,1H),7.39(m,3H),7.28-7.20(m,3H),7.10(br d,J=7.6Hz,1H),6.83(br d,J=7.8Hz,1H),5.34(s,2H),5.26(s,4H),3.59(br s,2H),2.42(brs,2H),1.66(br s,4H),1.24(br s,2H). ESI-MS m / z=443.2[M+H] + ;Calcu.=442.2.

[0297] Example 9: Synthesis of Compound A-9 / M-3

[0298]

[0299] Step A: Sodium borohydride (13.89 g, 367.2 mmol) was added to methanol (250 mL) dissolved with 3-hydroxy-4-methylbenzaldehyde (25 g, 183.6 mmol) at room temperature, and the whole mixture was stirred at 20°C for 16 hours. The reaction solution was poured into water (250 mL) at 0°C to quench, extracted with ethyl acetate (250 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated, and the residue was subjected to rapid silica gel chromatography ( 120g Silica flash lamp chromatography column, eluent 0-60% ethyl acetate / petroleum ether gradient @100 mL / min) gave 5-(hydroxymethyl)-2-methylphenol (9.7 g, 38%) as a yellow oil. 1 H NMR (400 MHz, DMSO-d 6 )δ9.14(s,1H),6.96(d,J=7.6Hz,1H),6.75(s,1H),6.61(d,J=7.5Hz,1H),5.01(t,J=5.8Hz,1H),4.36(d,J=5.8Hz,2H),2.07(s,3H).

[0300] Step B: At room temperature, add tert-butyldimethylsilyl chloride (10.58 g, 70.21 mmol, 8.64 mL) and imidazole (9.56 g, 140.4 mmol) to tetrahydrofuran (100 mL) in which 5-(hydroxymethyl)-2-methylphenol (9.7 g, 70.21 mmol) is dissolved. After the addition, the whole mixture is stirred at 25°C for 2 hours. The reaction solution is poured into water (100 mL), extracted with ethyl acetate (120 mL*3), the organic phase is dried over magnesium sulfate, filtered and concentrated. The residue is separated by silica gel column (SiO 2 , 0-9% petroleum ether:ethyl acetate, @100mL / min) to give a yellow oily product 5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methylphenol (9g, 51%). 1 H NMR (400 MHz, DMSO-d 6 )δ9.18(s,1H),6.97(d,J=7.6Hz,1H),6.75(s,1H),6.60(br d,J=7.5Hz,1H),4.57(s,2H),2.08(s,3H),0.89(s,9H),0.06(s,6H).

[0301] Step C: Add methyl 4-bromo-3-(methylsulfonyloxymethyl)benzoate (8 g, 24.76 mmol) and cesium carbonate (24.20 g, 74.27 mmol) to a solution of 5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methylphenol (6.56 g, 25.99 mmol) dissolved in acetonitrile (80 mL), and stir at 50 ° C for 16 hours. Pour the reaction mixture into water (150 mL), extract with ethyl acetate (150 mL*3), dry the organic phase with magnesium sulfate, filter and concentrate. The residue is 120g Silica flash chromatography column, eluent 0-9% petroleum ether: ethyl acetate, @100 mL / min, to give a yellow oily product 4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)benzoic acid methyl ester (9.5 g, 80%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.12(s,1H),7.79-7.89(m,2H),7.13(d,J=7.5Hz,1H),6.91(s,1H),6.81(br d,J=7.5Hz,1H),5.17(s,2H),4.65(s,2H),3.84(s,3H),2.21(s,3H),0.85(s,9H),0.02(s,6H).

[0302] Step D: Diisobutylaluminum hydride (1M, 98.2mL) was added to a solution of methyl 4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)benzoate (9.42g, 19.65mmol) dissolved in tetrahydrofuran (200mL), and the mixture was stirred at 0°C for 2 hours. The mixture was quenched with ice water (120mL), adjusted to pH 2 with dilute hydrochloric acid (1M), and extracted with ethyl acetate (40mL*3), and the organic layer was dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)phenyl)methanol (9g, crude product).

[0303] Step E: Triethylamine (6.05 g, 59.80 mmol, 8.32 mL) was added to a solution of (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)phenyl)methanol (9 g, crude) in dichloromethane (100 mL), and methanesulfonyl chloride (5.68 g, 49.58 mmol, 3.8 mL) was added at 0°C, and then stirred at 25°C for 2 hours. The mixture was poured into water (120 mL), extracted with dichloromethane (120 mL*3), and the organic phase was dried over anhydrous magnesium sulfate, filtered and concentrated to give a yellow oily product (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)phenyl)methyl methanesulfonate (10.6 g, crude).

[0304] Step F: To a solution of (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)phenyl)methyl methanesulfonate (10.6 g, 20.02 mmol) in dichloromethane (150 mL), 6-chloro-2H-pyrazolo[3,4-d]pyrimidin-4-amine (3.39 g, 20.02 mmol) and potassium carbonate (8.30 g, 60.05 mmol) were added and stirred at 25° C. for 4 hours. The mixture was filtered and concentrated to give a yellow oily product, 1-((4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)phenyl)methyl)-6-chloro-pyrazolo[3,4-d]pyrimidin-4-amine (12 g, crude). ESI-MS m / z=604.1[M+H] + ;Calcu.=603.1.

[0305] Step G: To a solution of compound 1-((4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)phenyl)methyl)-6-chloro-pyrazolo[3,4-d]pyrimidin-4-amine (12 g, crude product) in tetrahydrofuran (150 mL) was added tetrabutylammonium fluoride (1 M, 27.9 mL) and stirred at 25°C for 3 hours. The mixture was filtered and concentrated, and flash silica gel chromatography ( 120g Silica Flash column, eluent 0~50% tetrahydrofuran / petroleum ether gradient @100mL / min) to give a yellow solid product (3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenyl)methoxy)-4-methyl-phenyl)methanol (2g, 4.09mmol, four-step yield 21%). H NMR (400MHz, DMSO-d 6)δ8.33(br s,1H),8.20(br s,1H),8.09-8.14(m,1H),7.65(d,J=8.1Hz,1H),7.23-7.27(m,1H),7.20(dd,J=2.0,8.3Hz,1H),7.07(d,J=7.5Hz,1H),6.89 -6.94(m,1H),6.77-6.82(m,1H),5.47(s,2H),5.12(t,J=5.8Hz,1H),4.99–5.05(m,2H),4.43(d,J=5.8Hz,2H),2.02(s,3H).

[0306] Step H: Compound (3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenyl)methoxy)-4-methyl-phenyl)methanol (1g, 2.05mmol) in N-methylpyrrolidone (10mL) was added with sodium hydride (246mg, 6.14mmol, 60% content), and the mixture was stirred at 110°C for 8 hours. Then diluted with water (10mL), extracted with ethyl acetate (10mL*3), the organic layer was dried over magnesium sulfate, filtered and concentrated, and flash silica gel chromatography ( 20g Silica Flash column, eluent 0-50% tetrahydrofuran / petroleum ether gradient @80 mL / min), to obtain M-3, yellow solid product 3 4 -Br-6 6 -methyl-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (420 mg, 45%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.14-8.18(m,1H),7.89(s,1H),7.44-7.46(m,1H),7.08(dd,J=1.9,8.2Hz,1H),6. 97-6.92(m,1H),6.87(s,3H),6.76(d,J=7.5Hz,1H),5.38-5.31(m,6H),2.07(s,3H).

[0307] Step I: Sodium carbonate (70.30 mg, 663.3 μmol) was added to compound 3 4 -Br-6 6 -methyl-1 1H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (100 mg, 221.1 μmol) in tetrahydrofuran (0.6 mL) and water (0.3 mL) was added, followed by the addition of potassium trifluoro(pyrrolidin-1-ylmethyl)borate (42.2 mg, 221.1 μmol) and 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (10.5 mg, 22.11 μmol), and then under nitrogen protection, tris(dibenzylideneacetone)dipalladium (20.2 mg, 22.11 μmol) was added. The mixture was stirred at 80°C for 10 hours, then diluted with water (3 mL), extracted with ethyl acetate (3 mL*3), the organic layer was dried over magnesium sulfate, filtered and concentrated, and then purified by reverse phase high performance liquid chromatography (chromatographic column: C18 150*40mm*10um, mobile phase: [water (formic acid)-methanol], gradient: 8% to 48% B, 36 minutes) to obtain compound A-9, white solid product 6 6 -Methyl-3 4 -(pyrrolidin-1-ylmethyl)-1 1 H-5,8-Dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-1 4 -amine formate (25 mg, 25%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.20(s,1H),7.96(s,1H),7.90-7.85(m,1H),7.70-7.36(m,3H),7.26-7.20(m,1H),7.15-7.08(m,1H),6.94(br d,J=7.5Hz,1H),6.79-6.72(m,1H),5.45(s,2H),5.34(br d,J=3.9Hz,4H),3.75(br s,2H),2.45(br s,4H),2.03(s,3H),1.70(brs,4H). ESI-MS m / z=457.4[M+H] + ;Calcu.=456.2.

[0308] Example 10: Synthesis of Compound A-10 / M-4

[0309]

[0310] Step A: At room temperature, add tert-butyldimethylsilyl chloride (29.33 g, 194.6 mmol, 23.9 mL) and imidazole (26.50 g, 389.2 mmol) to tetrahydrofuran (300 mL) in which 5-(hydroxymethyl)-2-methoxyphenol (30 g, 194.6 mmol) is dissolved. After the addition, the mixture is stirred at 25°C for 2 hours. The reaction solution is filtered and concentrated, and the residue is subjected to rapid silica gel chromatography ( 120g Silica flash chromatography column, eluent 0-9% ethyl acetate / petroleum ether gradient @100 mL / min) gave a yellow oily product to give 5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxyphenol (30 g, 57%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.89(s,1H),6.84(d,J=8.3Hz,1H),6.74(d,J=2.1Hz,1H),6.66(dd,J=2.1,8.1Hz,1H),4.54(s,2H),3.73(s,3H),0.89(s,9H),0.05(s,6H).

[0311] Step B: At room temperature, add cesium carbonate (24.20 g, 74.27 mmol) and methyl 4-bromo-3-(methylsulfonyloxymethyl)benzoate (8 g, 24.76 mmol) to acetonitrile (80 mL) in which 5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxyphenol (6.31 g, 23.52 mmol) is dissolved. After the addition, the whole mixture is stirred at 50°C for 16 hours. The reaction solution is poured into water (150 mL) at 0°C, extracted with ethyl acetate (150 mL*3), and the organic phase is dried over magnesium sulfate, filtered, and concentrated. The residue was separated on a silica gel column (silica, 0-10% petroleum ether:ethyl acetate, @100 mL / min) to give a yellow oily product, 4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxy-phenoxy)methyl)benzoate (10.1 g, 82%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.11(s,1H),7.83(m,2H),6.95-7.00(m,1H),6.90-6.94(m,1H),6.84-6.89(m,1 H),5.16(s,2H),4.60(s,2H),3.85(s,3H),3.79(s,3H),0.85(s,9H),0.02(s,6H).

[0312] Step C: At 0°C, diisobutylaluminum hydride (1M, 101 mL) was added dropwise to a solution of 4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxy-phenoxy)methyl)benzoate (10 g, 20.18 mmol) in tetrahydrofuran (100 mL), and stirred at 0°C for 2 hours. The mixture was quenched with ice water (120 mL), adjusted to pH 2 with hydrochloric acid (1 M), and extracted with ethyl acetate (40 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated. A yellow oily product (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxy-phenoxy)methyl)phenyl)methanol (10.35 g, crude product) was obtained. 1 H NMR (400 MHz, DMSO-d 6 )δ7.60(d,J=8.1Hz,1H),7.52(d,J=1.6Hz,1H),7.24(dd,J=1.9,8.2Hz,1H),6.96(dd,J=3.3,4.9Hz,2H),6.88-6.83( m,1H),5.32(t,J=5.7Hz,1H),5.07(s,2H),4.61(s,2H),4.47(d,J=5.6Hz,2H),3.76(s,3H),0.87(s,9H),0.03(s,6H).

[0313] Step D: Triethylamine (6.72 g, 66.42 mmol, 9.3 mL) was added to a solution of the compound (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxy-phenoxy)methyl)phenyl)methanol (10.35 g, crude product) in dichloromethane (120 mL), and methanesulfonyl chloride (6.37 g, 55.61 mmol, 4.3 mL) was added at 0°C, and the mixture was stirred at 25°C for 2 hours. The mixture was diluted with water (120 mL), extracted with dichloromethane (20 mL*3), the organic layer was dried over magnesium sulfate, filtered and concentrated to obtain a yellow solid product (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxy-phenoxy)methyl)phenyl)methyl methanesulfonate (11.9 g, crude product).

[0314] Step E: 6-Chloro-2H-pyrazolo[3,4-d]pyrimidin-4-amine (3.70 g, 21.81 mmol) was added to a solution of compound (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxy-phenoxy)methyl)phenyl)methyl methanesulfonate (11.9 g, crude) in DMF (120 mL), and potassium carbonate (9.04 g, 65.44 mmol) was added, followed by stirring at 25° C. for 7 hours. The mixture was filtered and concentrated to give a yellow oily product, 1-((4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxyphenoxy)methyl)phenyl)methyl)-6-chloro-3a,7a-dihydropyrazolo[3,4-d]pyrimidin-4-amine (13.5 g, crude). ESI-MS m / z=620.1[M+H] + ;Calcu.=619.1.

[0315] Step F: To a solution of compound 1-((4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxyphenoxy)methyl)phenyl)methyl)-6-chloro-3a,7a-dihydropyrazolo[3,4-d]pyrimidin-4-amine (13.5 g, crude product) in tetrahydrofuran (150 mL) was added tetrabutylammonium fluoride (1 M, 30.55 mL) and stirred at 25° C. for 6 hours. The mixture was filtered and concentrated. The residue was purified by flash silica gel chromatography ( 120g Silica Flash column, eluent 0-68% tetrahydrofuran / petroleum ether gradient @100 mL / min) to give a yellow solid product (3-((5-((4-amino-6-chloro-3a,7a-dihydropyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenyl)methoxy)-4-methoxy-phenyl)methanol (4.1 g, four-step yield 40%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.16-8.36(m,2H),8.12(s,1H),7.63(d,J=8.3Hz,1H),7.37(d,J=1.6Hz,1H),7.15(dd,J=1.9,8.2Hz,1H),6.93-6.98(m ,1H),6.89-6.93(m,1H),6.64(s,1H),5.45(s,2H),5.04-5.08(m,1H),5.00-5.03(m,2H),4.36-4.41(m,2H),3.71(s,3H).

[0316] Step G: To a solution of compound (3-((5-((4-amino-6-chloro-3a,7a-dihydropyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenyl)methoxy)-4-methoxy-phenyl)methanol (1g, 1.98mmol) in N-methylpyrrolidone (10mL), sodium hydride (238mg, 5.94mmol, content 60%) was added and stirred at 110°C for 16 hours. The mixture was diluted with water (10mL) and extracted with ethyl acetate (10ml*2). The organic layer was dried over magnesium sulfate, filtered and concentrated, and flash silica gel chromatography ( 12g Silica Flash column, eluent 0-51% tetrahydrofuran / petroleum ether gradient @60 mL / min) to obtain M-4, orange solid product 3 4 -Br-6 6 -Methoxy-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (310 mg, 33%). ESI-MS m / z = 470.0 [M+H] + ;Calcu.=469.0.

[0317] Step H: Sodium carbonate (67.9 mg, 640.6 μmol) was added to compound 3. 4 -Br-6 6 -Methoxy-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (100 mg, 213.5 μmol) in tetrahydrofuran (0.6 mL) and water (0.3 mL) was added, and then potassium trifluoro(pyrrolidin-1-ylmethyl)borate (40.8 mg, 213.5 μmol) and 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (10.2 mg, 21.35 μmol) were added, and then tri(dibenzylideneacetone)dipalladium (19.6 mg, 21.35 μmol) was added under nitrogen protection. The mixed solution was stirred at 80°C for 10 hours, then diluted with water (3 mL), extracted with ethyl acetate (3 mL*3), the organic layer was dried over magnesium sulfate, filtered and concentrated, and then reversed-phase high performance liquid chromatography (chromatographic column: C18 150*40mm*10um, mobile phase: [water (formic acid)-methanol], gradient: 32%~72% B, 36min) to obtain compound A-10, a white solid product 6 6 -Methoxy-3 4 -(pyrrolidin-1-ylmethyl)-1 1H-5,8-Dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-1 4 -amine (7.25 mg, 31%). 1 H NMR (400 MHz, DMSO-d 6 )δ7.89(s,1H),7.33-7.76(m,4H),7.16-7.24(m,1H),7.08(br d,J=6.8Hz,1H),6.77-6.81(m,2H),5.38-5.42(m,2H),5.34(s,2H),5.28-5.31(m,2H),3.68(s,2H),3.63-3.66(m,3H),2.42(br d,J=3.8Hz,4H),1.66

[0318] -1.70(m,4H). ESI-MS m / z=473.4[M+H] + ;Calcu.=472.2.

[0319] Example 11: Synthesis of Compound A-11 / M-5

[0320]

[0321] Step A: Sodium borohydride (13.66 g, 361.1 mmol) was added to a methanol solution (250 mL) of 3-fluoro-5-hydroxy-benzaldehyde (25 g, 178.4 mmol). The mixture was stirred at 0°C for 2 hours. The mixture was concentrated to remove methanol, diluted with water (120 mL) at 0°C, extracted with ethyl acetate (120 mL*3), and dried over magnesium sulfate. Filtered and concentrated under reduced pressure to obtain 3-fluoro-5-(hydroxymethyl)phenol (19.58 g, 77%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ9.74(s,1H),6.58(s,1H),6.52(br d,J=9.8Hz,1H),6.39(td,J=2.3,10.9Hz,1H),5.23(br s,1H),4.41(s,2H).

[0322] Step B: Add tert-butyldimethylsilyl chloride (20.76 g, 137.8 mmol, 17 mL) and imidazole (18.76 g, 275.5 mmol) to a solution of 3-fluoro-5-(hydroxymethyl)phenol (19.58 g, 137.8 mmol) in tetrahydrofuran (200 mL) and stir at 25 ° C for 2 hours. The reaction mixture was quenched with water (200 mL) at 0 ° C. The reaction mixture was extracted with ethyl acetate (100 mL*3). Dry over magnesium sulfate, filter and concentrate under reduced pressure to obtain a white solid 3-(((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenol (29.5 g, 115.06 mmol, yield 84%). 1 H NMR (400 MHz, DMSO-d 6 )δ9.82(s,1H),6.58(s,1H),6.49(br d,J=9.6Hz,1H),6.40(td,J=2.1,10.9Hz,1H),4.62(s,2H),0.90(s,9H),0.07(s,6H).

[0323] Step C: Sodium borohydride (8.61 g, 227.6 mmol) was added in batches to a methanol solution (250 mL) of methyl 4-bromo-3-formylbenzoate (25 g, 102.86 mmol) and stirred at 0°C for 2 hours. The reaction solution was concentrated to remove methanol, diluted with water (120 mL) at 0°C, extracted with ethyl acetate (120 mL*3), and dried over magnesium sulfate. Filtered and concentrated under reduced pressure to obtain methyl 4-bromo-3-(hydroxymethyl)benzoate (18 g, 71%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ8.12(s,1H),7.57-7.77(m,2H),5.65(t,J=5.6Hz,1H),4.55(d,J=5.4Hz,2H),3.87(s,3H).

[0324] Step D: Add methyl 4-bromo-3-(hydroxymethyl)benzoate (18 g, 73.45 mmol), triethylamine (22.30 g, 220.4 mmol, 30.7 mL) to dichloromethane (200 mL), add methanesulfonyl chloride (21.23 g, 185.33 mmol, 14.34 mL) dropwise at 0°C, and stir at 25°C for 2 hours. The reaction solution was quenched with ice water (200 mL) at 0°C and extracted with dichloromethane (100 mL*2). Dry over magnesium sulfate, filter and concentrate under reduced pressure to obtain methyl 4-bromo-3-(chloromethyl)benzoate (19 g, 98%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6)δ=8.14(s,1H),7.86-7.91(m,2H),5.38(s,2H),3.88-3.87(m,3H).

[0325] Step E: Methyl 4-bromo-3-(chloromethyl)benzoate (8 g, 24.76 mmol) was dissolved in acetonitrile (80 mL), and cesium carbonate (24.20 g, 74.27 mmol) and 3-(((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenol (6.98 g, 27.23 mmol) were added and stirred at 50°C for 12 hours. The mixture was filtered and concentrated at 60°C and flash silica gel chromatography ( 80g Purification by silica flash lamp chromatography (0-15% ethyl acetate / petroleum ether gradient @ 100 mL / min) gave methyl 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)benzoate (9.2 g, 77%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ8.09(s,1H),7.82(d,J=0.9Hz,2H),6.77-6.85(m,2H),6.72(br d,J=9.9Hz,1H),5.17(s,2H),4.67(s,2H),3.84(s,3H),0.87(s,9H),0.04(s,6H).

[0326] Step F: Diisobutylaluminum hydride (1M, 95 mL) was added to a tetrahydrofuran solution (100 mL) of methyl 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)benzoate (9.2 g, 19.03 mmol). The mixture was stirred at 0°C for 2 hours. The mixture was quenched with ice water (200 mL), 1N hydrochloric acid was added to adjust the pH to 8, and the mixture was extracted with ethyl acetate (200 mL*2). The organic layer was dried over magnesium sulfate, filtered, and concentrated. A yellow oil (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl ester)methanol (10.63 g, crude product) was obtained.

[0327] Step G: To a solution of (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl ester)methanol (10.63 g, crude) in dichloromethane (100 mL), add methanesulfonyl chloride (6.69 g, 58.40 mmol, 4.5 mL) and triethylamine (4.72 g, 46.68 mmol, 6.50 mL), stir at 0°C for 2 hours, quench the reaction mixture with water (100 mL), and extract with dichloromethane (100 mL*3). Dry over magnesium sulfate, filter and concentrate under reduced pressure to give 4-bromo-3-((3-(((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl ester)benzylsulfonate (11.9 g, crude) as a yellow oil.

[0328] Step H: Potassium carbonate (9.25 g, 66.91 mmol), 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (4.54 g, 26.77 mmol) and 4-bromo-3-((3-(((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl)benzylsulfonate (11.9 g, crude) were added to N,N-dimethylformamide (100 mL) and stirred at 25°C for 12 hours. The reaction solution was filtered and the mother liquor was concentrated. 1-(4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl)benzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (15 g, crude) was obtained as a yellow solid. ESI-MS m / z=606.1[M+H] + ;Calcu.=605.1.

[0329] Step I: Add tetrabutylammonium fluoride (1M, 37.07mL, 1.5eq) to a solution of 1-(4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl ester)benzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (15g, crude) in tetrahydrofuran (130mL). Stir the reaction mixture at 25°C for 12 hours. Filter and concentrate the mother liquor. Use flash silica gel chromatography ( 80g Purification by silica flash lamp chromatography with a gradient of 0-40% tetrahydrofuran / petroleum ether @ 100 ml / min as eluent gave (3-((5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromobenzyl)oxy)-5-fluorophenyl)methanol (3.1 g, 6.29 mmol, 33% yield over four steps) as a white solid. ESI-MS m / z = 492.0 [M+H] + ;Calcu.=491.0

[0330] Step J: Add (3-((5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromobenzyl)oxy)-5-fluorophenyl)methanol (1 g, 2.03 mmol), sodium hydride (243.5 mg, 6.09 mmol, 60% purity) to a solution of N-methylpyrrolidone (100 mL). The mixture was stirred at 110°C for 2 hours. The reaction mixture was extracted with ethyl acetate (100 mL*2) and water (50 mL). The organic layers were combined, washed with brine (50 mL*2), dried over magnesium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The product was purified by flash silica gel chromatography ( 20g Purification by silica flash lamp chromatography (0-60% THF / petroleum ether gradient @ 100 ml / min) afforded M-5 as a white solid 3 4 -Br-6 5 -Fluoro-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (360 mg, 39%). ESI-MS m / z=456.0[M+H] + ;Calcu.=455.0.

[0331] Step K: Add 3 4 -Br-6 5 -Fluoro-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (100 mg, 219.17 μmol), potassium trifluoro(pyrrolidine-1-methyl)boranoate (37.67 mg, 197.2 μmol), dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (10.4 mg, 21.92 μmol), tri(dibenzylideneacetone)dipalladium (20.1 mg, 21.92 μmol) and sodium carbonate (69.7 mg, 657.5 μmol), purged with nitrogen three times, and then stirred under a nitrogen atmosphere at 80°C for 12 hours. The reaction mixture was extracted with ethyl acetate (10 mL*2) and water (10 mL). The organic layers were combined, washed with brine (50 mL*2), dried over magnesium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The crude product was purified by reverse phase high performance liquid chromatography (chromatographic column: xtimg C18 150*40mm*10um mobile phase: [water (formic acid)-methanol] gradient: 8% to 48% B, 36 minutes) to obtain compound A-11 as a white solid 6 5 -Fluoro-34 -(pyrrolidin-1-ylmethyl)-1 1 H-5,8-Dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-1 4 -amine (4.3 mg, 4%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.32-8.42(m,1H),7.83-7.97(m,2H),7.45-7.81(m,2H),7.43(s,1H),7.18-7.27(m,1H),7.0 5-7.16(m,1H),6.49-6.63(m,2H),5.45(s,2H),5.37(s,2H),5.33(s,2H),3.65(s,2H),2.42(br s,4H),1.69(br s,4H). ESI-MS m / z=461.2[M+H] + ;Calcu.=460.2.

[0332] Example 12: Synthesis of Compound A-12

[0333]

[0334] Step I: Dissolve 3 4 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (20 mg, 45.63 μmol) in tetrahydrofuran (0.4 mL), water (0.2 mL), potassium (morpholin-4-yl)methyl trifluoroborate (14.2 mg, 68.45 μmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (2.2 mg, 4.56 μmol), cesium carbonate (44.6 mg, 136.9 μmol) and palladium acetate (1.0 mg, 4.56 μmol) were added. After the addition, the flask was evacuated and nitrogen was injected for ventilation 3 times, and the whole mixture was stirred at 80 ° C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. Then, it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (FA)-MeOH]; gradient: 6%-46% B in 36 minutes) to obtain compound A-12, a white solid product 3 4 -(Morpholinemethyl)-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4-amine bisformate (3.89 mg, 19%). 1 HNMR (400 MHz, DMSO-d 6 )δ8.27(br s,2H),7.91(br s,2H),7.53(br s,3H),7.18(br s,1H),7.02-7.14(m,2H),6.84(br d,J=7.3Hz,1H),6.70(br d,J=7.1Hz,1H),5.44(br s,2H),5.29-5.40(m,4H),3.85-3.99(m,2H),3.17(s,4H),2.33(br s,4H). ESI-MS m / z=459.2[M+H] + ;Calcu.=458.2.

[0335] Example 13: Synthesis of Compound A-13 / A-16

[0336]

[0337] Step A: At 0°C, sodium borohydride (1.87 g, 49.37 mmol) was added to methanol (100 mL) in which methyl 4-bromo-3-formylbenzoate (10 g, 41.14 mmol) was dissolved. After the addition, the whole mixture was stirred at 0°C for 2 hours. The reaction solution was poured into ice water (100 mL), extracted with ethyl acetate (100 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. A yellow oily product, methyl 4-bromo-3-(hydroxymethyl)benzoate (10 g, 99%), was obtained. 1 H NMR (400 MHz, DMSO-d 6 )δppm8.10-8.14(m,1H),7.70-7.75(m,2H),5.65(t,J=5.6Hz,1H),4.55(d,J=5.6Hz,2H),3.87(s,3H).

[0338] Step B: At 0°C, add thionyl chloride (116.51 g, 979.3 mmol, 71.1 mL) to chloroform (240 mL) in which methyl 4-bromo-3-(hydroxymethyl)benzoate (24.00 g, 97.93 mmol) was dissolved. After the addition, the whole mixture was stirred at 80°C for 16 hours. The reaction solution was poured into ice water (250 mL), extracted with ethyl acetate (250 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated by silica gel column (silica, 0-10% petroleum ether: ethyl acetate, @100 mL / min) to obtain a white solid product methyl 4-bromo-3-(chloromethyl)benzoate (17 g, 66%). 1H NMR (400 MHz, DMSO-d 6 ) δppm8.20 (d, J = 1.3Hz, 1H), 7.82-7.88 (m, 2H), 4.92 (s, 2H), 3.88 (s, 3H).

[0339] Step C: At room temperature, 3-(((tert-butyldimethylsilyl)oxy)methyl)phenol (15.38 g, 64.51 mmol) and cesium carbonate (12.98 g, 39.85 mmol) were added to acetonitrile (17 mL) in which methyl 4-bromo-3-(chloromethyl)benzoate (17 g, 64.51 mmol) was dissolved. After the addition, the whole mixture was stirred at 50 ° C for 16 hours. The reaction solution was poured into ice water (250 mL), extracted with ethyl acetate (150 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated by silica gel column (silica, 0-4% petroleum ether: ethyl acetate, @100 mL / min) to obtain a yellow oily product 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester (3.2 g, 52%). 1 H NMR (400 MHz, DMSO-d 6 )δppm8.05(s,1H),7.23(t,J=7.8Hz,1H),7.79(s,2H),6.84-6.92(m,3H),5.13(s,2H),4.64(s,2H),3.80(s,3H),0.83(s,10H),0.00(s,6H).

[0340] Step D: At 0°C, diisobutylaluminum hydride (1M, 214.85 mL) was added to tetrahydrofuran (200 mL) in which 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester (20 g, 42.97 mmol) was dissolved. After the addition, the whole mixture was stirred at 0°C for 2 hours. The reaction solution was poured into ice water (300 mL), extracted with ethyl acetate (300 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. A white oily product (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)phenyl)methanol (18 g, 96%) was obtained. 1 H NMR (400 MHz, DMSO-d 6)δ7.55(d,J=8.1Hz,1H),7.41-7.50(m,1H),7.15-7.25(m,2H),6.89(br s,1H),6.81-6.87(m,2H),5.26(t,J=5.6Hz,1H),5.03(s,2H),4.63(s,2H),4.41(d,J=5.6Hz,2H),0.85-0.80(m,9H),0.02--0.03(m,6H).

[0341] Step E: At 0°C, methanesulfonyl chloride (7.39 g, 64.51 mmol, 4.99 mL) and triethylamine (6.94 g, 68.58 mmol, 9.55 mL) were added to dichloromethane (100 mL) in which (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)phenyl)methanol (10 g, 22.86 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into ice water (100 mL), extracted with dichloromethane (100 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. The yellow oily product 4-bromo-3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzylsulfonate (11 g, 93%) was obtained. 1 H NMR (400 MHz, DMSO-d 6 )δ7.68(d,J=8.3Hz,1H),7.57-7.61(m,1H),7.29-7.35(m,1H),7.22(t,J=7.8Hz,1H),6.89-6.91(m,1H),6.85(br d,J=7.6Hz,2H),5.20(s,2H),5.04-5.07(m,2H),4.63(s,2H),3.16(s,3H),0.83(s,9H),0.00(s,6H).

[0342] Step F: At room temperature, 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (3.62 g, 21.34 mmol) and potassium carbonate (8.85 g, 64.01 mmol) were added to N,N-dimethylformamide (250 mL) in which 4-bromo-3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzylsulfonate (11 g, 21.34 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 16 hours. The reaction solution was poured into water (500 mL) and extracted with ethyl acetate (500 mL*3). The organic phase was washed with brine and dried over magnesium sulfate, filtered and concentrated. The yellow oily product 1-(4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (12.5 g, 99%) was obtained. ESI-MS m / z=589.8[M+H] + ;Calcu.=589.1.

[0343] Step G: At 0°C, tetrabutylammonium fluoride (1M, 31.6 mL) was added to tetrahydrofuran (160 mL) in which 1-(4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (12.4 g, 21.05 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 h. The reaction solution was poured into ice water (160 mL), extracted with ethyl acetate (160 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated by silica gel column (silica, 0-41% petroleum ether: ethyl acetate, @100 mL / min) to obtain a yellow solid product (3-((5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromobenzyl)oxy)phenyl)methanol (4 g, 40%). ESI-MS m / z=475.8[M+H] + ;Calcu.=475.0.

[0344] Step H: At room temperature, sodium hydride (379 mg, 9.48 mmol, 60% purity) was added to 1-methylpyrrolidin-2-one (20 mL) in which (3-((5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromobenzyl)oxy)phenyl)methanol (1.5 g, 3.16 mmol) was dissolved. After the addition, the whole mixture was stirred at 110°C for 2 hours. The reaction solution was poured into ice water (160 mL), extracted with ethyl acetate (160 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. A-6, a yellow solid product 3, was obtained. 4 -Bromo-11 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctan-14-amine (600 mg, 43%). ESI-MS m / z=439.9[M+H] + ;Calcu.=439.0.

[0345] Step I: Dissolve 3 4 -Bromo-1 1 To a solution of H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctan-14-amine (30 mg, 68.45 μmol) in tetrahydrofuran (0.4 mL) were added water (0.2 mL), potassium trifluoro((4-methylpiperazin-1-yl)methyl)borate (15.1 mg, 68.45 μmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (3.3 mg, 6.84 μmol), tris(dibenzylideneacetone)dipalladium (6.3 mg, 6.84 μmol) and sodium carbonate (21.8 mg, 205.35 μmol). After the addition was complete, the flask was evacuated and then ventilated with nitrogen three times. The whole mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. Then, it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 6%-46% B in 36 minutes) to obtain compound A-13, a white solid product 3 4 -((4-methylpiperazin-1-yl)methyl)-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine formate (5.08 mg, 15%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.16(s,1H),7.88-7.92(m,2H),7.53(m,3H),7.16(d,J=7.9Hz,1H),7.04-7.09(m,2H),6.83(d,J=7.4Hz,1H),6. 66-6.70(m,1H),5.43(s,2H),5.35(s,2H),5.33(s,2H),3.51(s,2H),2.45-2.47(m,4H),2.32(s,3H),2.17(m,4H). ESI-MS m / z=472.1[M+H] + ;Calcu.=471.2.

[0346] Example 14: Synthesis of Compound A-14

[0347]

[0348] Step A: Dissolve 3 4 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (30 mg, 68.45 μmol) in tetrahydrofuran (0.4 mL), water (0.2 mL), potassium (dimethylamino)methyl trifluoroborate (11.3 mg, 68.45 μmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (3.3 mg, 6.84 μmol), tris(dibenzylideneacetone)dipalladium (6.3 mg, 6.84 μmol) and sodium carbonate (21.8 mg, 205.35 μmol) were added. After the addition, the flask was evacuated and nitrogen was injected for ventilation 3 times, and the whole mixture was stirred at 80 ° C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. Then, it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 4%-44% B in 36 minutes) to obtain compound A-14, a white solid product 3 4 -((dimethylamino)methyl)-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine formate (8.32 mg, 28%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.20(s,1H),7.88-7.94(m,2H),7.53(m,3H),7.15-7.21(m,1H),7.07(dd,J=8.2,16.1Hz,2H),6 .84(d,J=7.5Hz,1H),6.67(dd,J=1.8,8.1Hz,1H),5.41(s,2H),5.36(s,2H),5.34(s,2H),3.44(br s,2H),2.14(s,6H). ESI-MS m / z=417.2[M+H] + ;Calcu.=416.2.

[0349] Example 15: Synthesis of Compound A-15

[0350]

[0351] Step A: Sodium borohydride (0.45 g, 11.9 mmol) was added in batches to methanol (160 mL) in which methyl 4-fluoro-3-formylbenzoate (6.0 g, 32.9 mmol) was dissolved at 0°C. After the addition, the whole mixture was stirred at 25°C for 2 hours. Water (100 mL) was added to the reaction solution, and extracted with ethyl acetate (200 mL*3) for 3 times. The organic phase was dried over magnesium sulfate, filtered and concentrated to obtain methyl 4-fluoro-3-(hydroxymethyl)benzoate (5.90 g, 97%). 1 H NMR: (400MHz, DMSO-d 6 )δppm 8.12(dd,J=2.2,7.2Hz,1H),7.84-7.97(m,1H),7.30(dd,J=8.7,9.8Hz,1H),5.47(t,J=5.8Hz,1H),4.60(d,J=5.8Hz,2H),3.86(s,3H). ESI-MS m / z=184.7[M+H] + ;Calcu.=184.1.

[0352] Step B: At 0°C, triethylamine (9.56 g, 94.5 mmol) and methanesulfonyl chloride (5.70 g, 49.8 mmol) were added to dichloromethane (100 mL) in which methyl 4-fluoro-3-(hydroxymethyl)benzoate (5.80 g, 31.5 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into water (50 mL), extracted with dichloromethane (100 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product, methyl 4-fluoro-3-(methylsulfonyloxymethyl)benzoate (5.60 g, crude product).

[0353] Step C: At room temperature, 3-(((tert-butyldimethylsilyl)oxy)methyl)phenol (5.60 g, 23.5 mmol) and cesium carbonate (10.4 g, 32.0 mmol) were added to a solution of 4-fluoro-3-(methylsulfonyloxymethyl)benzoic acid methyl ester (5.60 g, crude product) in acetonitrile (100 mL), and the mixture was stirred at 50°C for 16 hours. The reaction solution was filtered and concentrated. The product was purified by flash silica gel chromatography ( 80g The mixture was purified by silica gel chromatography with 0-3% ethyl acetate / petroleum ether as the eluent to give a yellow oily product, 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-4-fluorobenzoic acid methyl ester (5.81 g, two-step yield 46%).

[0354] Step D: At 0°C, diisobutylaluminum hydride (1M, 57.4 mL) was added to tetrahydrofuran (100 mL) in which methyl 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-4-fluorobenzoate (5.80 g, 14.3 mmol) was dissolved. The mixture was stirred at 0°C for 2 hours. The mixture was quenched with ice water (100 mL) and hydrochloric acid (1N, 50 mL), extracted with ethyl acetate (100 mL*3), and the organic layer was dried over magnesium sulfate, filtered and concentrated to give a yellow oily product, 3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-4-fluorophenyl)methanol (5.38 g, crude product). 1 H NMR: (400MHz, DMSO-d 6 )δppm 7.41(dd,J=1.9,7.2Hz,1H),7.23-7.30(m,1H),7.16-7.23(m,1H),7.12(dd,J=8.6,10.1Hz,1H),6.89(s,1H),6.8 7-6.80(m,2H),5.21(t,J=5.7Hz,1H),5.04(s,2H),4.62(s,2H),4.41(d,J=5.6Hz,2H),0.83(s,9H),0.00(s,6H).

[0355] Step E: At 0°C, triethylamine (1.61 g, 15.9 mmol) and methanesulfonyl chloride (1.24 g, 10.8 mmol) were added to a solution of 3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-4-fluorophenyl)methanol (2.00 g, 5.31 mmol) in dichloromethane (40 mL). After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into water (30 mL), extracted with dichloromethane (50 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product tert-butyl-((3-((5-(chloromethyl)-2-fluoro-phenyl)methoxy)phenyl)formyl)-dimethylsilane (2.55 g, crude product).

[0356] Step F: At room temperature, tert-butyl-((3-((5-(chloromethyl)-2-fluoro-phenyl)methoxy)phenyl)formyl)-dimethylsilane (1.00 g, 2.53 mmol) and potassium carbonate (1.05 g, 7.60 mmol) were added to N,N-dimethylformamide (20 mL) in which 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (429 mg, 2.53 mmol) was dissolved, and the mixture was stirred at 25°C for 16 hours. The mixture was diluted with water (10 mL), extracted with ethyl acetate (20 mL*3), the organic layer was dried over magnesium sulfate, filtered and concentrated to give a yellow oily product (3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-4-fluorophenyl)methanol (1.30 g, crude product). ESI-MS m / z=528.3[M+H] + ;Calcu.=527.2.

[0357] Step G: At 0°C, tetrabutylammonium fluoride (1M, 3.6 mL) was added to tetrahydrofuran (15 mL) in which (3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-4-fluorophenyl)methanol (1.27 g, 2.40 mmol) was dissolved, and the mixture was stirred at 25°C for 3 hours. The mixture was diluted with water (10 mL), extracted with ethyl acetate (20 ml*3), and the organic layer was dried over magnesium sulfate, filtered, and concentrated. Flash silica gel chromatography ( 20g Silica gel chromatography, eluent 0-47% ethyl acetate / petroleum ether) to obtain a yellow solid product (3-((5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-fluorobenzyl)oxy)phenyl)methanol (430 mg, 43%). ESI-MS m / z=413.9[M+H] + ;Calcu.=413.1.

[0358] Step H: At 0°C, sodium hydride (43.5 mg, 1.09 mmol, content 60%) was added to a solution of (3-((5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-fluorobenzyl)oxy)phenyl)methanol (150 mg, 362 μmol) dissolved in N-methylpyrrolidone (3 mL), and stirred at 110°C for 2 hours. The mixture was filtered and then purified by preparative high performance liquid chromatography (formic acid conditions, chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 32%-72% B in 36 minutes) to obtain compound A-15, a white solid product 3 4 -Fluoro-1 1H-5,8-Dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-1 4 -amine (5.10 mg, 4%). 1 H NMR: (400MHz, DMSO-d 6 )δppm 8.14(dd,J=1.6,7.1Hz,1H),7.94(s,2H),7.57(s,2H),7.16-7.23(m,1H),7.01-7.15(m,2H) ,6.88(d,J=7.6Hz,1H),6.70(dd,J=2.1,8.1Hz,1H),5.42(s,2H),5.36(s,2H),5.33(s,2H). ESI-MS m / z=378.1[M+H] + ;Calcu.=377.1.

[0359] Example 16: Synthesis of Compound A-17

[0360]

[0361] Step A: Add 3-formyl-4-hydroxy-benzoic acid methyl ester (10 g, 55.51 mmol), potassium carbonate (11.51 g, 83.26 mmol) and iodomethane (9.45 g, 66.61 mmol, 4.15 mL) to N,N-dimethylformamide (200 mL), and stir the reaction solution at 25 ° C for 12 hours. Pour the reaction solution into water (500 mL) and ethyl acetate (500 mL*2), and separate the upper and lower layers. The organic layer is washed with 500 mL*2 brine, and the aqueous layers are combined and extracted with ethyl acetate. The organic layer is dried over magnesium sulfate, filtered, and concentrated in vacuo to obtain 3-formyl-4-methoxy-benzoic acid methyl ester (12 g, crude product) as a yellow oil. 1 HNMR (400 MHz, DMSO-d 6 )δ10.35(s,1H),8.16-8.31(m,2H),7.38(d,J=8.8Hz,1H),4.01(s,3H),3.85(s,3H).

[0362] Step B: At 0°C, sodium borohydride (2.81 g, 74.16 mmol) was added to a methanol solution (120 mL) of 3-formyl-4-methoxy-benzoic acid methyl ester (12 g, crude product), and stirred at 0°C for 2 hours. The reaction solution was diluted with water (120 mL) at 0°C, extracted with ethyl acetate (120 mL*3), and dried over magnesium sulfate. Filtered and concentrated under reduced pressure to obtain a yellow oily substance, 3-(hydroxymethyl)-4-methoxybenzoic acid methyl ester (7.69 g, two-step yield 70%).1 H NMR (400 MHz, DMSO-d 6 )δ7.98–8.09(m,1H),7.87(dd,J=2.3,8.6Hz,1H),7.07(d,J=8.6Hz,1H),5.22(t,J=5.7Hz,1H),4.50(d,J=5.6Hz,2H),3.86(s,3H),3.82(s,3H).

[0363] Step C: At 0°C, methanesulfonyl chloride (4.4 g, 38.41 mmol, 3.0 mL) was added to a solution of 3-(hydroxymethyl)-4-methoxy-benzoic acid methyl ester (3 g, 15.29 mmol) and triethylamine (3.09 g, 30.58 mmol, 4.3 mL) in dichloromethane (50 mL). The mixture was stirred at 25°C for 2 hours. The reaction solution was quenched with water (100 mL) at 0°C. The reaction mixture was extracted with dichloromethane (100 mL). Drying over magnesium sulfate, filtering and concentrating under reduced pressure gave 3-(chloromethyl)-4-methoxybenzoic acid methyl ester (2.7 g, 82%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ8.02(d,J=2.1Hz,1H),7.97(dd,J=2.3,8.6Hz,1H),7.17(d,J=8.8Hz,1H),4.77(s,2H),3.93(s,3H),3.83(s,3H).

[0364] Step D: Add 3-(chloromethyl)-4-methoxy-benzoic acid methyl ester (2.2 g, 10.25 mmol), 3-(((tert-butyldimethylsilyl)oxy)methyl)phenol (2.69 g, 11.27 mmol) and cesium carbonate (10.02 g, 30.75 mmol) to acetonitrile (80 mL). Stir the mixture at 50°C for 12 hours. Filter the mixture and concentrate the mother liquor at 50°C. Use flash silica gel chromatography ( 40g Silica flash lamp chromatography with a gradient of 0-20% ethyl acetate / petroleum ether @ 100 ml / min as the eluent gave methyl 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl)-4-methoxybenzoate (3.82 g, 89%) as a yellow oil. 1 H NMR (400 MHz, DMSO-d 6)δ7.90–8.00(m,2H),7.30-7.14(m,2H),6.82-6.94(m,3H),5.09(s,2H),4.67(s,2H),3.91(s,3H),3.80(s,3H),0.87(s,9H),0.04(s,6H).

[0365] Step E: Diisobutylaluminum hydride (1M, 21.84 mL) was added to a tetrahydrofuran solution (20 mL) of methyl 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl)-4-methoxybenzoate (1.82 g, 4.37 mmol). The mixture was stirred at 0°C for 2 hours. The reaction solution was quenched with ice water (50 mL), 1N hydrochloric acid was added to pH = 8, and ethyl acetate (50 mL*3) was used for extraction. The organic layer was dried over magnesium sulfate, filtered and concentrated. A white solid (3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl)-4-methoxyphenyl)methanol (1.6 g, 94%) was obtained. 1 H NMR (400 MHz, DMSO-d 6 )δ7.34(d,J=1.8Hz,1H),7.21-7.28(m,2H),6.99(d,J=8.4Hz,1H),6.93(s,1H),6.90-6.82(m,2H),5.08( t,J=5.7Hz,1H),5.03(s,2H),4.68(s,2H),4.42(d,J=5.6Hz,2H),3.80(s,3H),0.89(s,9H),0.06(s,6H).

[0366] Step F: Methanesulfonyl chloride (1.46 g, 12.75 mmol, 986 μL) was added to a solution of (3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-4-methoxyphenyl)methanol (1.6 g, 4.12 mmol) and triethylamine (1.25 g, 12.35 mmol, 1.7 mL) in dichloromethane (50 mL) at 0°C. Stir at 25°C for 2 hours. The reaction mixture was quenched with water (50 mL) at 0°C. The reaction mixture was extracted with dichloromethane (50 mL*3). Dry over magnesium sulfate, filter and concentrate under reduced pressure to give (3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-4-methoxyphenyl)methanesulfonic acid methyl ester (1.0 g, 52%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6)δ7.46-7.51(m,1H),7.41(dd,J=1.9,8.4Hz,1H),7.19-7.26(m,1H),7.05(d,J=8.4Hz,1H),6.92-6.98(m,1H), 6.86-6.91(m,2H),5.02(s,2H),4.75(s,2H),4.47(s,2H),3.84(s,3H),2.37(s,3H),0.84(s,9H),-0.04(s,6H).

[0367] Step G: To a solution of (3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-4-methoxyphenyl)methanesulfonic acid methyl ester (200 mg, 683.2 μmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (115.8 mg, 683.2 μmol) in N,N-dimethylformamide (4 mL), potassium carbonate (188.8 mg, 1.37 mmol) was added and stirred at 25°C for 12 hours. The reaction solution was filtered and the mother liquor was concentrated at 60°C to give 1-((3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-4-methoxy-phenyl)methyl)-6-chloropyrazolo[3,4-d]pyrimidin-4-amine (0.36 g, crude) as a yellow oil. ESI-MS m / z=540.5[M+H] + ;Calcu.=539.2.

[0368] Step H: To a solution of 1-((3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-4-methoxy-phenyl)methyl)-6-chloropyrazolo[3,4-d]pyrimidin-4-amine (360 mg, 666.5 μmol) in tetrahydrofuran (20 mL), add tetrabutylammonium fluoride (1 M, 799.8 μL) and stir at 25°C for 2 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL*2). The combined organic layers were dried, filtered and concentrated under reduced pressure to give a crude product. The product was purified by flash silica gel chromatography ( 4g Silica Flash column, eluent 0-27% tetrahydrofuran / petroleum ether gradient @100 mL / min) purification, to obtain the compound (3-((5-((4-amino-6-chloropyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxy-phenyl)methoxy)phenyl)methanol (71 mg, crude) as a white solid. ESI-MS m / z=426.3[M+H] + ;Calcu.=425.1.

[0369] Step I: To a solution of (3-((5-((4-amino-6-chloropyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxy-phenyl)methoxy)phenyl)methanol (66 mg, 155.0 μmol) in N-methylpyrrolidone (2 mL) at 0°C, sodium hydride (18.60 mg, 464.93 μmol, 60% purity) was added. The mixture was stirred at 110°C for 2 hours. The reaction mixture was extracted with ethyl acetate (10 mL*2) and water (10 mL). The combined organic layers were washed with brine (10 mL*2), dried over magnesium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase high performance liquid chromatography (chromatographic column: C18 150*40 mm*10 um mobile phase: [water (formic acid)-methanol] gradient: 30%-70% B, 36 minutes) to obtain compound A-17 as a white solid 3 4 -Methoxy-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (4 mg, three-step yield 1.5%). 1 H NMR (400MHz, CD 3 OD)δ8.08(s,1H),7.91(d,J=2.3Hz,1H),7.57(s,1H),7.25(dd,J=2.3,8.6Hz,1H),7.11-7.19(m,1H),6.98(d,J=7 .6Hz,1H),6.86(d,J=8.5Hz,1H),6.78(dd,J=2.1,8.1Hz,1H),5.60(s,2H),5.44(s,2H),5.29(s,2H),3.80(s,3H). ESI-MS m / z=390.2[M+H] + ;Calcu.=389.2.

[0370] Example 17: Synthesis of Compound A-18

[0371]

[0372] Step A: At room temperature, potassium carbonate (12.61 g, 91.24 mmol) and 3-(bromomethyl)benzaldehyde (9.08 g, 45.62 mmol) were added to acetonitrile (100 mL) in which 5-(hydroxymethyl)-2-methoxyphenol (7.03 g, 45.62 mmol) was dissolved. The mixture was stirred at 50°C for 5 hours. The reaction solution was filtered and concentrated, and the residue was subjected to flash silica gel chromatography ( 40g Silica Flash column, eluting with 0-35% ethyl acetate / petroleum ether gradient @100 mL / min) to give 3-((5-(hydroxymethyl)-2-methoxy-phenoxy)methyl)benzaldehyde (6.8 g, 55%) as a yellow oil. 1 HNMR (400 MHz, DMSO-d 6 )δ10.04(s,1H),7.99(s,1H),7.89(br d,J=7.6Hz,1H),7.79(br d,J=7.5Hz,1H),7.60-7.68(m,1H),7.04(s,1H),6.91-6.97(m,1H),6.85-6.90 (m,1H),5.17(s,2H),5.07(t,J=5.7Hz,1H),4.40(d,J=5.6Hz,2H),3.76(s,3H).

[0373] Step B: At room temperature, triethylamine (7.58 g, 74.92 mmol, 10.43 mL) was added to dichloromethane (100 mL) in which 3-((5-(hydroxymethyl)-2-methoxy-phenoxy)methyl)benzaldehyde (6.8 g, 24.97 mmol) was dissolved, and methanesulfonyl chloride (5.82 g, 50.81 mmol, 3.93 mL) was added dropwise at 0°C. After the addition, the whole mixture was stirred at 25°C for 4 hours. The reaction solution was poured into water (120 mL), extracted with ethyl acetate (120 mL*3), and the organic phase was dried over anhydrous magnesium sulfate, filtered and concentrated. A white solid product (3-((3-formylphenyl)methoxy)-4-methoxy-phenyl)methyl methanesulfonate (5.26 g, crude product) was obtained.

[0374] Step C: Add compound 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.20 g, 7.05 mmol) and potassium carbonate (3.08 g, 22.26 mmol) to a solution of (3-((3-formylphenyl)methoxy)-4-methoxy-phenyl)methyl methanesulfonate (2.6 g, 7.42 mmol) in N,N-dimethylformamide (30 mL), and stir at 40°C for 5 hours. Filter and concentrate the mixture. 20g Silica Flash column, eluent 0-27% petroleum ether:tetrahydrofuran, @100mL / min), to obtain a yellow solid product 3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxy-phenoxy)methyl)benzaldehyde (1.3 g, 3.07 mmol, two-step yield 25%). ESI-MS m / z=424.1[M+H] +;Calcu.=423.1.

[0375] Step D: Sodium borohydride (0.250 g, 6.61 mmol) was added to a solution of compound 3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxy-phenoxy)methyl)benzaldehyde (0.8 g, 1.89 mmol) in methanol (8 mL), and the mixture was stirred at 0°C for 10 hours. The mixture was quenched with ice water (10 mL), extracted with ethyl acetate (40 mL*3), and the organic layer was dried over magnesium sulfate, filtered and concentrated to give a yellow solid product (3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxy-phenoxy)methyl)phenyl)methanol (700 mg, 87%). ESI-MS m / z=426.1[M+H] + ;Calcu.=425.1.

[0376] Step E: Sodium hydride (28.2 mg, 704.4 μmol, content 60%) was added to a solution of compound (3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxy-phenoxy)methyl)phenyl)methanol (100 mg, 234.81 μmol) in N-methylpyrrolidone (2 mL) and stirred at 110° C. for 3 hours. The mixture was poured into water (1 mL), extracted with ethyl acetate (1 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated, and then purified by reverse phase high performance liquid chromatography (chromatographic column: C18 150*40mm*10um, mobile phase: [water (formic acid)-methanol], gradient: 26% to 66% B, 36 minutes) to obtain a white solid product 3. 4 -Methoxy-1 1 H-4,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (12 mg, 13%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.02(s,1H),7.86(s,1H),7.35-7.67(m,3H),7.20-7.28(m,3H),6.72-6.87(m,2H),5.35(s,2H),5.22(s,4H),3.65(s,3H). ESI-MS m / z=390.4[M+H] + ;Calcu.=389.2.

[0377] Example 18: Synthesis of Compound A-19 / A-20

[0378]

[0379] Step A: Dissolve 3 4 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (200mg, 456.33μmol) in methanol (100mL) and N,N-dimethylformamide (5mL), add 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (33.4mg, 45.63μmol) and triethylamine (92.4mg, 912.7μmol, 127μL). After the addition, the whole mixture was stirred at 100°C for 16 hours under a carbon monoxide (1MPa) atmosphere. Water (200mL) was added to the reaction solution, and extracted with ethyl acetate (200mL*3) three times. The organic phase was dried over magnesium sulfate, filtered and concentrated. Then, it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (FA)-MeOH]; gradient: 32%-72% B in 36 minutes) to obtain a white solid product A-20, 1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-3 4 -carboxylic acid methyl ester formate (1.41 mg, 1%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.47(br s,1H),8.07(s,1H),7.93(s,1H),7.62(d,J=7.8Hz,2H),7.49(s,2H),7.21(d,J=1.0Hz,1H),7.09-7.04(m,1H),6.85(d,J=7.0Hz,1H),6.70(br d,J=7.8Hz,1H),5.56(s,2H),5.43(s,2H),5.32(s,2H),3.84(s,3H). ESI-MS m / z=418.1[M+H] + ;Calcu.=417.1.

[0380] Step B: At 0°C, dissolve 1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-3 4-Methyl carboxylate (50 mg, 119.78 μmol) in tetrahydrofuran (0.4 mL) was added with sodium hydroxide (19.2 mg, 479.2 μmol) and water (0.1 mL). After the addition, the whole mixture was stirred at 25 ° C for 1 hour. Ethyl acetate (2 mL) was added to the reaction solution and extracted 3 times with water (2 mL * 3). The reaction mixture was concentrated under reduced pressure to remove the solvent. Then it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150 * 40 mm * 10 um; mobile phase: [water (formic acid) - methanol]; gradient: 22% -62% B in 36 minutes) to obtain compound A-19, a white solid product 1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-3 4 -carboxylic acid (13.48 mg, 27%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.01(br s,1H),7.92(s,1H),7.62(br d,J=7.9Hz,2H),7.54(s,1H),7.39-7.51(m,1H),7.20(br d,J=7.8Hz,1H),7.05-7.13(m,1H),6.86(d,J=7.6Hz,1H),6.73(br d,J=7.1Hz,1H),5.61(s,2H),5.40(s,2H),5.33(s,2H). ESI-MS m / z=404.1[M+H] + ;Calcu.=403.1.

[0381] Example 19: Synthesis of Compound A-22 / M-6

[0382]

[0383] Step A: Dissolve 3 4 -Bromo-1 1 H-4,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4-amine (200mg, 456.3μmol) in methanol (5mL), add N,N-dimethylformamide (5mL), 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (33.4mg, 45.63μmol) and triethylamine (92.3mg, 912.6μmol, 127μL). After the addition, the whole mixture was stirred at 100°C for 48 hours under a carbon monoxide (1MPa) atmosphere. The reaction solution was poured into water (10mL), extracted with ethyl acetate (10mL*3), and the organic phase was dried with magnesium sulfate, filtered and concentrated. M-6, a yellow solid product 1, was obtained. 4 -Amino-1 1 H-4,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzoylcyclooctane-3 4 -Methyl carboxylate (200 mg, crude product). ESI-MS m / z=418.2[M+H] + ;Calcu.=417.1.

[0384] Step B: At 0°C, dissolve 1 4 -Amino-1 1 H-4,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzoylcyclooctane-3 4 -Methyl carboxylate (100 mg, 239.6 μmol) in tetrahydrofuran (1 mL) was added with diisobutylaluminum hydride (1M, 1.2 mL). After the addition, the whole mixture was stirred at 0°C for 2 hours. The reaction solution was poured into water (1 mL), extracted with ethyl acetate (1 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. Then, it was purified by reverse phase high performance liquid chromatography (chromatographic column: chromatographic column: C18 150*30mm*5um, mobile phase: [water (formic acid)-acetonitrile], gradient: 2%-42% B over25min) to obtain compound A-22, a white solid product (1 4 -Amino-1 1 H-4,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-3 4 -yl)methanol (5.12 mg, two-step yield 6%). 1 H NMR (400 MHz, DMSO-d 6)δ8.03(s,1H),7.88(s,1H),7.38(s,3H),7.22-7.27(m,3H),7.15(d,J=7.8Hz,1H),6.85(d,J=8. 6Hz, 1H), 5.34 (s, 2H), 5.26 (d, J = 6.1Hz, 4H), 4.90 (t, J = 5.7Hz, 1H), 4.41 (d, J = 5.3Hz, 2H); ESI-MS m / z=390.1[M+H] + ;Calcu.=389.2.

[0385] Example 20: Synthesis of Compound A-23

[0386]

[0387] Step A: 3-(((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenol (10.30 g, 40.19 mmol), potassium carbonate (11.11 g, 80.38 mmol) and 3-(bromomethyl)benzaldehyde (8 g, 40.19 mmol) were added to N,N-dimethylformamide (100 mL) and stirred at 25 °C for 12 hours. The reaction was poured into water (500 mL) and ethyl acetate (500 mL*2) and the layers were separated. The organic layer was washed with 500 mL*2 brine and the combined aqueous layers were extracted with ether. The organic layers were combined, dried over magnesium sulfate, filtered, and concentrated in vacuo to give 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl ester)benzaldehyde (12 g, 80%) as a yellow oil. 1 H NMR (400 MHz, DMSO-d 6 )δ9.79-10.19(m,1H),7.97(s,1H),7.81-7.91(m,1H),7.77(br d,J=7.6Hz,1H),7.56-7.69(m,1H),6.74-7.04(m,2H),6.70(br d,J=9.3Hz,1H),5.22(s,2H),4.67(s,2H),0.83-0.94(m,9H),0.14-0.20(m,6H).

[0388] Step B: To a tetrahydrofuran solution (150 mL) of 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl)benzaldehyde (11.5 g, 30.71 mmol), tetrabutylammonium fluoride (1M, 46 mL) was added, and the reaction solution was stirred at 25°C for 12 hours. The reaction solution was quenched with water (100 mL) at 0°C. The reaction mixture was extracted with ethyl acetate (100 mL*3). Drying over magnesium sulfate, filtering and concentrating under reduced pressure gave 3-((3-fluoro-5-(hydroxymethyl)phenoxy)methyl)benzaldehyde (7.2 g, 90%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ10.04(s,1H),7.98(s,1H),7.89(d,J=7.6Hz,1H),7.78(d,J=7.6Hz,1H),7.60-7.68(m,1H ), 6.85 (s, 1H), 6.68-6.81 (m, 2H), 5.32 (t, J = 5.8Hz, 1H), 5.22 (s, 2H), 4.48 (d, J = 5.9Hz, 2H).

[0389] Step C: At 0°C, triethylamine (7.00 g, 69.16 mmol, 9.6 mL) and 3-((3-fluoro-5-(hydroxymethyl)phenoxy)methyl)benzaldehyde (7.2 g, 27.66 mmol) were added to dichloromethane (100 mL), the temperature was lowered to 0°C, methanesulfonyl chloride (8.22 g, 71.76 mmol, 5.6 mL) was added, and the reaction solution was stirred at 25°C for 2 hours. The reaction solution was quenched with water (100 mL) at 0°C. The reaction mixture was extracted with dichloromethane (100 mL*3). Drying over magnesium sulfate, filtering and concentrating under reduced pressure gave methyl 3-(chloromethyl)-4-methoxybenzoate (10.8 g, crude product) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ10.03(s,1H),7.99(s,1H),7.89(d,J=7.6Hz,1H),7.79(br d,J=7.6Hz,1H),7.59-7.68(m,1H),7.00(s,1H),6.86-6.98(m,2H),5.25(s,2H),5.23(s,2H).

[0390] Step D: To a solution of 3-(chloromethyl)-4-methoxy-benzoic acid methyl ester (2 g, crude) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (952 mg, 5.62 mmol) in N,N-dimethylformamide (20 mL) was added potassium carbonate (2.45 g, 17.73 mmol). The mixture was stirred at 25 °C for 12 hours. The reaction solution was filtered and the mother liquor was concentrated. The residue was purified by flash silica gel chromatography ( 40g Silica flash lamp chromatography with a gradient of 0-40% tetrahydrofuran / petroleum ether @ 100 ml / min as eluent gave 3-((3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenoxy)methyl as a white solid (700 mg, 1.70 mmol, 33% yield over two steps). ESI-MS m / z = 412.1 [M+H] + ;Calcu.=411.1.

[0391] Step E: To a solution of 3-((3-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenoxy)methyl (300 mg, 728.5 μmol) in tetrahydrofuran / methanol=2:1 (10 mL) at 0°C, sodium borohydride (55.1 mg, 1.46 mmol) was added in portions. The mixture was stirred at 0°C for 2 hours. Diluted with water (50 mL) at 0°C, extracted with ethyl acetate (50 mL*3), and dried over magnesium sulfate. Filtration and concentration under reduced pressure gave a white solid (3-((3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenoxy)methylphenyl)methanol (400 mg, crude), ESI-MS m / z=414.1[M+H] + ;Calcu.=413.1.

[0392] Step F: Sodium hydride (29.0 mg, 724.9 μmol, 60% purity) was added to a solution of (3-((3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenoxy)methylphenyl)methanol (100 mg, crude) in N-methylpyrrolidone (2 mL) at 0°C. The mixture was stirred at 110°C for 3 hours. The reaction solution was quenched with water (2 mL) and extracted with ethyl acetate (5 mL*2). The organic layers were combined, washed with brine (5 mL*2), dried over magnesium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. It was then purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (FA)-MeOH]; gradient: 36%-76% B, over 36 min) to obtain compound A-23 as a white solid 3 5 -Fluoro-1 1 H-4,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-1 4 -amine (5.21 mg, 7% yield in two steps). 1 H NMR (400 MHz, DMSO-d 6 )δ8.45(s,1H),8.04(s,1H),7.91(s,1H),7.51 -7.75(m,1H),7.46 -7.50(m,1H),7.34(s,1H),7.26(s,2H),6.54 -6.61(m,2H),5.35(s,2H),5.31(s,2H),5.26(s,2H). ESI-MS m / z=378.3[M+H] + ;Calcu.=377.1.

[0393] Example 21: Synthesis of Compound A-24

[0394]

[0395] Step A: Add a mixture of 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (8.51 g, 11.63 mmol) and triethylamine (23.53 g, 232.51 mmol, 32.36 mL) to a solution of 3-bromo-5-methoxy-benzaldehyde (25 g, 116.26 mmol) in methanol (300 mL), degas and purify with nitrogen three times, and then stir at 80°C for 12 hours under a carbon monoxide atmosphere. The reaction solution was diluted with water (200 mL) and extracted with ethyl acetate (220 mL). The organic phase was dried over magnesium sulfate, filtered and concentrated, and the residue was subjected to rapid silica gel chromatography ( 80g The residue was purified by chromatography on a silica gel column with 0-8% ethyl acetate / petroleum ether as the eluent to obtain 3-formyl-5-methoxy-benzoic acid methyl ester (21.6 g, 96%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ10.05(s,1H),8.05(d,J=1.1Hz,1H),7.73-7.66(m,2H),3.87-3.89(m,6H). ESI-MS m / z=194.9[M+H] + ,Calcu.=194.1.

[0396] Step B: Sodium borohydride (30.09 g, 795.35 mmol) was added to a methanol solution (210 mL) of 3-formyl-5-methoxy-benzoic acid methyl ester (21.5 g, 110.72 mmol) and stirred at 0°C for 4 hours. The reaction solution was poured into water (120 mL), extracted with ethyl acetate (120 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. A white oily product, 3-(hydroxymethyl)-5-methoxy-benzoic acid methyl ester (20 g, 101.94 mmol, yield 92%), was obtained. 1 H NMR (400 MHz, DMSO-d 6 )δ7.53(s,1H),7.31(s,1H),7.15(s,1H),5.35(t,J=5.8Hz,1H),4.53(d,J=5.9Hz,2H),3.84(s,3H),3.80(s,3H). ESI-MS m / z=196.9[M+H] + ;Calcu.=196.1.

[0397] Step C: To a solution of 3-(hydroxymethyl)-5-methoxy-benzoic acid methyl ester (10 g, 50.97 mmol) in carbon tetrachloride (100 mL), add thionyl chloride (60.64 g, 509.7 mmol, 37 mL) and stir at 80°C for 16 hours. The mixture is filtered and concentrated. Flash silica gel chromatography ( 40g The residue was purified by chromatography on a silica gel column with 0-3% ethyl acetate / petroleum ether as the eluent to obtain 3-(chloromethyl)-5-methoxy-benzoic acid methyl ester (5.3 g, 48%) as a yellow oily product. 1 H NMR (400 MHz, DMSO-d 6)δ7.63(s,1H),7.41(dd,J=1.4,2.3Hz,1H),7.35-7.26(m,1H),4.81(s,2H),3.86(s,3H),3.82(s,3H). ESI-MS m / z=214.9[M+H] + ;Calcu.=214.0.

[0398] Step D: Cesium carbonate (23.68 g, 72.68 mmol) and compound 3-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (5.78 g, 24.23 mmol) were added to a solution of 3-(chloromethyl)-5-methoxy-benzoic acid methyl ester (5.2 g, 24.23 mmol) in acetonitrile (50 mL), and the mixture was stirred at 50°C for 16 hours. The mixture was filtered and concentrated. Flash silica gel chromatography ( 120 g Silica column chromatography, eluent 0-5% ethyl acetate / petroleum ether gradient @100 ml / min) gave 3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-5-methoxy-benzoic acid methyl ester (7.5 g, 74%) as a colorless oily product. 1 H NMR (400 MHz, DMSO-d 6 )δ7.62(s,1H),7.36-7.43(m,1H),7.21-7.29(m,2H),6.84-6.95(m,3H),5.16( s,2H),4.66(s,2H),3.85(s,3H),3.81(s,3H),0.87(s,9H),0.08-0.02(m,6H).

[0399] Step E: At 0°C, diisobutylaluminum hydride (1M, 48 mL) was added to a solution of 3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-5-methoxy-benzoic acid methyl ester (4 g, 9.60 mmol) / tetrahydrofuran (80 mL), and then stirred at 0°C for 2 hours. The mixture was quenched with ice water (80 mL), extracted with ethyl acetate (10 mL*3), and the organic layer was dried over magnesium sulfate, filtered and concentrated to give a yellow oily product (3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-5-methoxy-phenyl)methanol (3.6 g, 93%). 1 H NMR (400 MHz, DMSO-d 6)δ7.21-7.28(m,1H),6.94(br d,J=11.3Hz,2H),6.80-6.90(m,4H),5.22(t,J=5.8Hz,1H),5.04(s,2H),4 .67(s,2H),4.47(d,J=5.6Hz,2H),3.74(s,3H),0.89(s,9H),0.06(s,6H).

[0400] Step F: At 0°C, methanesulfonyl chloride (1.38 g, 12.05 mmol, 932.43 μL) was added to a solution of (3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-5-methoxy-phenyl)methanol (2 g, 5.15 mmol) and triethylamine (1.56 g, 15.44 mmol, 2.15 mL) in dichloromethane (40 mL) and stirred at 25°C for 1 hour. The mixture was quenched with water (30 mL) at 0°C, extracted with ethyl acetate (30 mL*3), and the organic layer was dried over magnesium sulfate, filtered, and concentrated. The mixture was purified by flash silica gel chromatography ( 12g Silica Flash column, eluent: 0-45% ethyl acetate / petroleum ether gradient @60 mL / min) to give the product (3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-5-methoxy-phenyl)methyl methanesulfonate (900 mg, 1.93 mmol, yield 58%) as a yellow oil. 1 H NMR (400 MHz, DMSO-d 6 )δ7.22-7.27(m,1H),7.09(s,1H),7.02-7.04(m,1H),6.97(s,1H),6.93(d,J=0.9Hz,1H),6.88(dd,J=2.0,7.9H z,2H),5.23(s,2H),5.06-5.10(m,2H),4.67(s,2H),3.76-3.78(m,3H),3.22(s,3H),0.89(s,9H),0.06(s,6H).

[0401] Step G: To a solution of the compound tert-butyl-((3-((3-(chloromethyl)-5-methoxy-phenyl)methoxy)phenyl)formyl)-dimethylsilane (200 mg, 491.4 μmol) in N,N-dimethylformamide (2 mL), potassium carbonate (204 mg, 1.47 mmol) and 6-chloro-2H-pyrazolo[3,4-d]pyrimidin-4-amine (100 mg, 589.7 μmol) were added and stirred at 25°C for 4 hours. The mixture was filtered and concentrated to give a yellow oily product 2-((3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-5-methoxy-phenyl)methyl)-6-chloro-pyrazolo[3,4-d]pyrimidin-4-amine (1.0 g, crude). ESI-MS m / z=540.5[M+H] + ;Calcu.=539.2.

[0402] Step H: Tetrabutylammonium fluoride (1M, 2.22mL) was added to a tetrahydrofuran (10mL) solution of compound 2-((3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-5-methoxy-phenyl)methyl)-6-chloro-pyrazolo[3,4-d]pyrimidin-4-amine (1g, crude product), and then stirred at 25°C for 2 hours. The mixture was diluted with water (10mL), extracted with ethyl acetate (10mL*3), the organic layer was dried over magnesium sulfate, filtered and concentrated, and flash silica gel chromatography ( 12g Silica Flash column, eluting with 0-54% ethyl acetate / petroleum ether gradient @60 mL / min) to give the product (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-2-yl)methyl)-5-methoxy-phenyl)methoxy)phenyl)methanol (260 mg) as a yellow oil. 1 H NMR (400 MHz, DMSO-d 6 )δ8.15-8.36(m,2H),8.12(s,1H),7.21(t,J=7.8Hz,1H),6.93(s,2H),6.87(s,1H),6.78-6.84(m,2H ), 6.73 (s, 1H), 5.42 (s, 2H), 5.16 (t, J = 5.8Hz, 1H), 5.00 (s, 2H), 4.45 (d, J = 5.8Hz, 2H), 3.72 (s, 3H). ESI-MSm / z=426.3[M+H] + ;Calcu.=425.1.

[0403] Step I: At 0°C and under nitrogen protection, sodium hydride (45.1 mg, 1.13 mmol, 60% purity) was added to a solution of (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-2-yl)methyl)-5-methoxy-phenyl)methoxy)phenyl)methanol (160 mg, 375.7 μmol) in N-methylpyrrolidone (2 mL), and stirred at 100°C for 2 hours. The mixed solution was diluted with water (2 mL), extracted with ethyl acetate (2 mL*3), the organic layer was dried over magnesium sulfate, filtered and concentrated, and the crude product was purified by reverse phase high performance liquid chromatography (chromatographic column: C18 150*40mm*10um, mobile phase: [water (FA)-MeOH], gradient: 32% to 72% B, 36 min). Compound A-24, white solid product 3, was obtained. 5 -Methoxy-1 1 H-5,8-Dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctan-14-amine (15 mg, 38.52 μmol, yield 5%). 1 H NMR (400MHz, CD 3 OD)δ8.01(s,1H),7.46(s,1H),7.41(s,1H),7.12(t,J=7.9Hz,1H),6.95(s,1H),6.86(s,1H), 6.75(dd,J=2.0,8.6Hz,1H),6.66(s,1H),5.48(s,2H),5.44(s,2H),5.20(s,2H),3.64(s,3H). ESI-MS m / z=390.2[M+H] + ;Calcu.=389.2.

[0404] Example 22: Synthesis of Compound A-25

[0405]

[0406] Step A: Sodium borohydride (4.06 g, 107 mmol, 1.63 eq) was added to methanol (100 mL) at 0°C in which 3-hydroxy-5-methoxy-benzaldehyde (10.0 g, 65.7 mmol, 1 eq) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. Water (100 mL) was added to the reaction solution, and it was extracted three times with ethyl acetate (200 mL*3). The organic phase was dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product, 3-(hydroxymethyl)-5-methoxyphenol (11.1 g, crude product).

[0407] Step B: At room temperature, add tert-butyldimethylsilyl chloride (9.78 g, 64.87 mmol, 7.98 mL, 1 eq) and imidazole (13.25 g, 194.60 mmol, 3 eq) to tetrahydrofuran (110 mL) in which 3-(hydroxymethyl)-5-methoxyphenol (10 g, 64.87 mmol, 1 eq) is dissolved. After the addition, the whole mixture is stirred at 25°C for 16 hours. The reaction solution is poured into water (100 mL), extracted with ethyl acetate (100 mL*3), the organic phase is dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product 3-((tert-butyl(dimethyl)silyl)oxymethyl)-5-methoxy-phenol (12.4 g, 71%). 1 H NMR: (400MHz, DMSO-d 6 )δppm9.28(s,1H),6.25(br d,J=10.6Hz,2H),6.12(t,J=2.1Hz,1H),4.51(s,2H),3.60(s,3H),0.84(s,9H),0.00(s,6H).

[0408] Step C: At room temperature, add 3-((tert-butyl(dimethyl)silyl)oxymethyl)-5-methoxy-phenol (12.4g, 46.20mmol, 1eq) and cesium carbonate (45.15g, 138.59mmol, 3eq) to acetonitrile (80mL) in which 3-(bromomethyl)benzaldehyde (11.03g, 55.43mmol, 1.2eq) was dissolved, and stirred at 50°C for 16 hours. Pour the reaction solution into water (100mL), extract with ethyl acetate (100mL*3), dry the organic phase with magnesium sulfate, filter and concentrate. Use flash silica gel chromatography ( 80g Silica gel chromatography column, eluent 0-6% ethyl acetate / petroleum ether) to obtain a yellow oily product 3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)-5-methoxy-phenoxy)methyl)benzaldehyde (6.59 g, 37%). ESI-MS m / z=387.2[M+H] + ;Calcu.=386.2.

[0409] Step D: At 0°C, tetrabutylammonium fluoride (1M, 25.57 mL, 1.5 eq) was added to THF (100 mL) in which 3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)-5-methoxy-phenoxy)methyl)benzaldehyde (6.59 g, 17.05 mmol, 1 eq) was dissolved, and the mixture was stirred at 25°C for 2 hours. The mixture was diluted with 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 Silica gel chromatography column, eluent 0-40% ethyl acetate / petroleum ether) to obtain a yellow oily product 3-((3-(hydroxymethyl)-5-methoxy-phenoxy)methyl)benzaldehyde (3.78 g, yield 75%). ESI-MS m / z=273.1[M+H] + ;Calcu.=272.1.

[0410] Step E: At 0°C, triethylamine (4.21 g, 41.65 mmol, 5.80 mL, 3 eq) and methanesulfonyl chloride (2.630 g, 22.96 mmol, 1.78 mL, 1.65 eq) were added to dichloromethane (60 mL) in which 3-((3-(hydroxymethyl)-5-methoxy-phenoxy)methyl)benzaldehyde (3.78 g, 13.88 mmol, 1 eq) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into water (50 mL), extracted with dichloromethane (60 mL*3), the organic phase was dried over magnesium sulfate, and concentrated to give a yellow oily product (3-((3-formylphenyl)methoxy)-5-methoxy-phenyl)methyl methanesulfonate (4.62 g, crude product).

[0411] Step F: At room temperature, 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2.24 g, 13.19 mmol, 1 eq) and potassium carbonate (3.64 g, 26.37 mmol, 2 eq) were added to N,N-dimethylformamide (80 mL) in which (3-((3-formylphenyl)methoxy)-5-methoxy-phenyl)methyl methanesulfonate (4.62 g, crude product) was dissolved, and the mixture was stirred at 25°C for 16 hours. The mixture was filtered and concentrated, and the residue was purified by flash silica gel chromatography ( 80g Silica gel chromatography column, eluent 0-32% ethyl acetate / petroleum ether) to obtain a yellow solid 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxy-phenoxy)methyl)benzaldehyde (2.2 g, two-step yield 28%). ESI-MS m / z=424.1[M+H] + ;Calcu.=423.1.

[0412] Step G: Sodium borohydride (30 mg, 792.97 μmol, 3.36 eq) was added to tetrahydrofuran (3 mL) in which 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxy-phenoxy)methyl)benzaldehyde (100 mg, 235.93 μmol, 1 eq) was dissolved at 0°C, and the mixture was stirred at 25°C for 2 hours. The mixture was diluted with water (5 mL), extracted with ethyl acetate (5 mL*3), and the organic layer was dried over magnesium sulfate, filtered and concentrated to give a yellow solid product (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxy-phenoxy)methyl)phenyl)methanol (110 mg, crude product). ESI-MS m / z=426.1[M+H] + ;Calcu.=425.1

[0413] Step H: At 0°C, sodium hydride (30.99 mg, 774.89 μmol, 60% content, 3 eq) was added to a solution of (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxy-phenoxy)methyl)phenyl)methanol (110 mg, 258.3 μmol, 1 eq) in N-methylpyrrolidone (3 mL), and stirred at 110°C for 2 hours. After the mixture was filtered, it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 34%-74% B in 36 minutes) to obtain compound A-25, a white solid product 3 5 -Methoxy-1 1 H-4,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (5.53 mg, yield 5%). 1 H NMR: (400MHz, DMSO-d 6 )δppm 8.00(s,1H),7.89(s,1H),7.78-7.32(m,2H),7.25(s,3H),7.05(s,1H),6. 41 (s, 1H), 6.26 (s, 1H), 5.35 (s, 2H), 5.22 (d, J = 14.3Hz, 4H), 3.58 (s, 3H). ESI-MS m / z=390.2[M+H] + ;Calcu.=389.2

[0414] Example 23: Synthesis of Compound A-26 / M-7

[0415]

[0416] Step A: Sodium borohydride (1.87 g, 49.37 mmol) was added in batches to methanol (25 mL) / tetrahydrofuran (250 mL) in which dimethyl 5-bromobenzene-1,3-dicarboxylate (50 g, 183.10 mmol) was dissolved at 0°C. After the addition, the whole mixture was stirred at 0°C for 2 hours. The reaction solution was poured into ice water (50 mL), extracted with ethyl acetate (100 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated. A white oily product, methyl 3-bromo-5-(hydroxymethyl)benzoate (40 g, 89%), was obtained. 1 H NMR (400 MHz, DMSO-d 6 )δ7.91-7.92(m,2H),7.78(s,1H),5.50(t,J=5.8Hz,1H),4.57(d,J=5.8Hz,2H),3.87(s,3H).

[0417] Step B: At 0°C, add triethylamine (37.16 g, 367.2 mmol, 51.1 mL) and methanesulfonyl chloride (29.26 g, 255.4 mmol, 19.8 mL) to dichloromethane (500 mL) in which 3-bromo-5-(hydroxymethyl)benzoic acid methyl ester (30 g, 122.4 mmol) is dissolved. After the addition, the whole mixture is stirred at 25°C for 2 hours. The reaction solution is poured into ice water (500 mL), extracted with ethyl acetate (500 mL*3), and the organic phase is dried over anhydrous magnesium sulfate, filtered and concentrated. A yellow oily product, 3-bromo-5-(methylsulfonyloxymethyl)benzoic acid methyl ester (35 g, crude product) is obtained.

[0418] Step C: At room temperature, cesium carbonate (60.49 g, 185.7 mmol) and 3-(((tert-butyldimethylsilyl)oxy)methyl)phenol (14.75 g, 61.89 mmol) were added to acetonitrile (200 mL) in which methyl 3-bromo-5-(methylsulfonyloxymethyl)benzoate (20 g, crude product) was dissolved. After the addition, the whole mixture was stirred at 50°C for 16 hours. The reaction solution was poured into ice water (200 mL), extracted with ethyl acetate (200 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated by silica gel column (SiO 2 , 0~5%, petroleum ether:ethyl acetate, @100mL / min) to give a yellow oily product, 3-bromo-5-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)benzoic acid methyl ester (24g, two-step yield 74%). 1 H NMR (400 MHz, DMSO-d 6)δ7.94-7.98(m,2H),7.82-7.93(m,1H),7.22(t,J=7.8Hz,1H),6.81-6.92 (m,3H),5.16(s,2H),4.62(s,2H),3.82(s,3H),0.83(s,9H),0.00(s,6H).

[0419] Step D: At 0°C, diisobutylaluminum hydride (1M, 215 mL) was added to tetrahydrofuran (200 mL) in which methyl 3-bromo-5-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)benzoate (20 g, 42.97 mmol) was dissolved. After the addition, the whole mixture was stirred at 0°C for 2 hours. The reaction solution was poured into ice water (200 mL), extracted with ethyl acetate (200 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. A white oily product [3-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (18.5 g, 98%) was obtained. 1 HNMR (400 MHz, DMSO-d 6 )δ7.38-7.41(m,1H),7.32(s,1H),7.18(d,J=8.0Hz,2H),6.80-6.89(m,3H),5.30(t,J= 5.8Hz,1H),5.04(s,2H),4.61(s,2H),4.44(d,J=5.8Hz,2H),0.83(s,9H),0.00(s,6H).

[0420] Step E: At 0°C, triethylamine (12.84 g, 126.9 mmol, 17.7 mL) and methanesulfonyl chloride (10.94 g, 95.50 mmol, 7.4 mL) were added to dichloromethane (200 mL) in which (3-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (18.5 g, 42.29 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into ice water (100 mL), extracted with dichloromethane (100 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. The yellow oily product 3-bromo-5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzylsulfonate (20 g, crude product) was obtained.

[0421] Step F: At room temperature, potassium carbonate (10.72 g, 77.59 mmol) and 6-chloro-2H-pyrazolo[3,4-d]pyrimidin-4-amine (9.21 g, 54.31 mmol) were added to N,N-dimethylformamide (200 mL) in which 3-bromo-5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzylsulfonate (20 g, crude product) was dissolved. After the addition, the whole mixture was stirred at 25°C for 16 hours. The reaction solution was poured into water (200 mL) and extracted with ethyl acetate (200 mL*3). The organic phase was washed with brine and dried over magnesium sulfate, filtered and concentrated. The yellow oily product 1-(3-bromo-5-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (22.6 g, crude product) was obtained. ESI-MS m / z=590.1[M+H] + ;Calcu.=589.1

[0422] Step G: At 0°C, tetrabutylammonium fluoride (1M, 57.3 mL) was added to tetrahydrofuran (400 mL) in which 1-(3-bromo-5-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (22.5 g, crude product) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into ice water (200 mL), extracted with ethyl acetate (400 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated on a silica gel column (silica, 0-53% petroleum ether:ethyl acetate, @100 mL / min) to give a yellow solid product (3-((3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromobenzyl)oxy)phenyl)methanol (5 g, three-step yield 25%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.18-8.39(m,2H),8.11-8.18(m,1H),7.57(s,1H),7.38(s,1H),7.27(s,1H),7.18-7.25(m,1H),6.94(s,1H),6.90(br d,J=7.6Hz,1H),6.81(dd,J=2.1,8.1Hz,1H),5.48(s,2H),5.17(t,J=5.8Hz,1H),5.01-5.11(m,2H),4.46(d,J=5.8Hz,2H)

[0423] Step H: At room temperature, sodium hydride (505 mg, 12.64 mmol, 60% purity) was added to N-methylpyrrolidone (20 mL) in which (3-((3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-bromobenzyl)oxy)phenyl)methanol (2 g, 4.21 mmol) was dissolved. After the addition, the whole mixture was stirred at 110°C for 2 hours. The reaction solution was poured into ice water (200 mL), extracted with ethyl acetate (200 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. M-7, a yellow solid product 3, was obtained. 5 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (0.5 g, yield 27%). ESI-MS m / z = 440.0 [M+H] + ;Calcu.=439.0.

[0424] Step I: Dissolve 3 5 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (50 mg, 114.08 μmol) in dioxane (1 mL) and water (0.2 mL). Potassium hydroxide (12.80 mg, 228.2 μmol) and methanesulfonic acid (2-dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium (II) (10.3 mg, 11.41 μmol) were added, and the mixture was stirred at 80°C for 3 hours. Water (1 mL) was added to the reaction solution, and extracted 3 times with ethyl acetate (1 mL*3). The organic phase was dried over magnesium sulfate, filtered and concentrated, and then purified by reverse phase high performance liquid chromatography (chromatographic column: chromatographic column: C18 150*40mm*10um, mobile phase: [water (formic acid)-methanol], gradient: 24% to 64% B, 36min) to obtain compound A-26, a white solid product 1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-3 5 -alcohol (1 mg, 2%). 1 H NMR (400 MHz, DMSO-d 6)δ9.37(s,1H),7.84-7.95(m,1H),7.42-7.51(m,2H),7.37-7.41(m,1H),7.13-7.30(m,1H),7.03-7.11(m,1H),6.81-6.91 (m,1H),6.66-6.70(m,1H),6.59-6.63(m,1H),6.45-6.51(m,1H),5.32-5.35(m,2H),5.25-5.30(m,2H),5.10-5.15(m,2H). ESI-MS m / z=376.1[M+H] + ;Calcu.=375.1.

[0425] Example 24: Synthesis of Compound A-27

[0426]

[0427] Step A: Dissolve 3 5 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (50 mg, 114.08 μmol) in tetrahydrofuran (0.4 mL), water (0.2 mL), potassium trifluoro(pyrrolidin-1-ylmethyl)borate (21.8 mg, 114.08 μmol), sodium carbonate (36.27 mg, 342.2 μmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (5.4 mg, 11.41 μmol) and tris(dibenzylideneacetone)dipalladium (10.4 mg, 11.41 μmol) were added. After the addition, the flask was evacuated and nitrogen was injected for ventilation 3 times, and the whole mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. Then, it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 8%-48% B in 36 minutes) to obtain compound A-27, a white solid product 3 5 -(pyrrolidin-1-ylmethyl)-1 1 H-5,8-Dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzoylcyclooctane-1 4 -amine formate (5.36 mg, 10%). 1 H NMR (400 MHz, DMSO-d 6)δ8.27(s,1H),7.90(s,1H),7.85(s,1H),7.49(m,3H),7.20(s,1H),7.05-7.10(m,2H) ,6.85(d,J=7.1Hz,1H),6.68(dd,J=1.8,9.1Hz,1H),5.36(s,4H),5.22(s,2H),3.41(br s,2H),2.42-2.43(m,2H),1.62-1.63(m,4H),1.24(br s,2H). ESI-MS m / z=443.2[M+H] + ;Calcu.=442.2.

[0428] Example 25: Synthesis of Compound A-28

[0429]

[0430] Step A: Dissolve 3 5 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (50 mg, 114.08 μmol) in tetrahydrofuran (0.4 mL), water (0.2 mL), potassium trifluoro(morpholinemethyl)borate (23.6 mg, 114.1 μmol), sodium carbonate (36.3 mg, 342.3 μmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (5.4 mg, 11.41 μmol) and tris(dibenzylideneacetone)dipalladium (10.4 mg, 11.41 μmol) were added. After the addition, the flask was evacuated and nitrogen was injected for ventilation 3 times, and the whole mixture was stirred at 80 ° C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. Then, it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 6%-46% B in 36 minutes) to obtain compound A-28, a white solid product 3 5 -(Morpholinemethyl)-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine formate (6.51 mg, 12%). 1 H NMR (400 MHz, DMSO-d 6)δ8.28(s,1H),7.91(s,1H),7.86(s,1H),7.47(m,3H),7.20(s,1H),7.04-7.10(m,2H),6.85(d,J=7.5Hz,1H),6.68(br d,J=6.5Hz,1H),5.36(br d,J=3.6Hz,4H),5.23(s,2H),3.51(br s,2H),3.28(br s,4H),2.22(br s,4H). ESI-MS m / z=459.2[M+H] + ;Calcu.=458.2.

[0431] Example 26: Synthesis of Compound A-29

[0432]

[0433] Step A: Dissolve 3 5 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (50 mg, 114.08 μmol) in tetrahydrofuran (0.4 mL), water (0.2 mL), potassium trifluoro-((4-methylpiperazine-1-yl)methyl)borate (25.1 mg, 114.08 μmol), sodium carbonate (36.3 mg, 342.25 μmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (5.4 mg, 11.41 μmol) and tris(dibenzylideneacetone)dipalladium (10.4 mg, 11.41 μmol) were added. After the addition, the flask was evacuated and nitrogen was injected for ventilation 3 times, and the whole mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. Then, it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 6%-46% B in 36 minutes) to obtain compound A-29, a white solid product 3 5 -((4-methylpiperazin-1-yl)methyl)-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine formate (7.42 mg, 13%). 1 H NMR (400 MHz, DMSO-d 6)δ8.20(s,1H),7.90(s,1H),7.85(s,1H),7.48(br s,3H),7.19(s,1H),7.12-7.02(m,2H),6.85(d,J=7.1Hz,1H),6.70-6.66(m,1H),5.36(s,4H),5.22(s,2H),3.28(br s,2H),2.30-2.20(m,7H),2.14(s,4H). ESI-MS m / z=472.2[M+H] + ;Calcu.=471.2.

[0434] Example 27: Synthesis of Compound A-30

[0435]

[0436] Step A: Dissolve 3 5 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (50 mg, 114.08 μmol) in tetrahydrofuran (0.4 mL), water (0.2 mL), (dimethylamino) methyl trifluoroborate potassium (18.8 mg, 114.08 μmol), sodium carbonate (36.3 mg, 342.25 μmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (5.4 mg, 11.41 μmol) and tris(dibenzylideneacetone)dipalladium (10.4 mg, 11.41 μmol) were added. After the addition, the flask was evacuated and nitrogen was injected for ventilation 3 times, and the whole mixture was stirred at 80 ° C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. Then, it was purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 6%-46% B in 36 minutes) to obtain compound A-30, a white solid product 3 5 -((dimethylamino)methyl)-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine formate (5.62 mg, 11%). 1 H NMR (400 MHz, DMSO-d 6)δ8.29(s,1H),7.90(s,1H),7.86(s,1H),7.49(m,3H),7.19(s,1H),7.11-7.04(m,2H),6.8 5(d,J=7.8Hz,1H),6.70-6.65(m,1H),5.36(s,4H),5.22(s,2H),3.22(s,2H),2.04(s,6H). ESI-MS m / z=417.2[M+H] + ;Calcu.=416.2.

[0437] Example 28: Synthesis of Compound A-31

[0438]

[0439] Step A: Sodium borohydride (3.27 g, 86.44 mmol, 1.57 eq) was added to methanol (50 mL) and tetrahydrofuran (50 mL) at 0°C in which methyl 2-fluoro-5-formylbenzoate (10 g, 54.90 mmol, 1 eq) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. Water (100 mL) was added to the reaction solution, and it was extracted three times with ethyl acetate (200 mL*3). The organic phase was dried over magnesium sulfate, filtered and concentrated to obtain a colorless oily product, methyl 2-fluoro-5-(hydroxymethyl)benzoate (10.2 g, crude product).

[0440] Step B: At 0°C, triethylamine (16.81 g, 166.16 mmol, 23 mL) and methanesulfonyl chloride (9.85 g, 85.99 mmol, 6.7 mL) were added to dichloromethane (150 mL) in which methyl 2-fluoro-5-(hydroxymethyl)benzoate (10.2 g, crude product) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into water (50 mL), extracted with dichloromethane (100 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product, methyl 2-fluoro-5-(((methylsulfonyl)oxy)methyl)benzoate (12 g, crude product).

[0441] Step C: At room temperature, 3-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (7.77 g, 32.57 mmol) and cesium carbonate (28.95 g, 88.84 mmol) were added to acetonitrile (100 mL) in which methyl 2-fluoro-5-(((methylsulfonyl)oxy)methyl)benzoate (6 g, crude product) was dissolved, and the mixture was stirred at 50°C for 16 hours. The reaction solution was poured into water (100 mL), extracted with ethyl acetate (150 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product, 5-((3-(((dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-benzoic acid methyl ester (3.47 g, crude product).

[0442] Step D: Diisobutylaluminum hydride (1M, 43mL) was added to tetrahydrofuran (30mL) in which 5-((3-(((dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-benzoic acid methyl ester (3.47g, crude product) was dissolved at 0°C. The mixture was stirred at 0°C for 2 hours. The mixture was quenched with ice water (50mL) and hydrochloric acid (1N, 10mL), extracted with ethyl acetate (50mL*3), and the organic layer was dried over magnesium sulfate, filtered and concentrated to give a yellow oily product (5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-phenyl)methanol (2.90g, 28% yield for four steps). 1 H NMR: (400MHz, DMSO-d 6 )δppm 7.44-7.54(m,1H),7.26-7.35(m,1H),7.19(t,J=7.9Hz,1H),7.08(dd,J=8.5,10.0Hz,1H),6.77-6.90(m ,3H),5.25(t,J=5.7Hz,1H),5.00(s,2H),4.61(s,2H),4.49(d,J=5.6Hz,2H),0.83(s,9H),0.00(s,6H).

[0443] Step E: At 0°C, triethylamine (2.38 g, 23.53 mmol, 3.3 mL) and methanesulfonyl chloride (1.68 g, 14.67 mmol, 1.14 mL) were added to dichloromethane (50 mL) in which (5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-phenyl)methanol (2.95 g, 7.84 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into water (30 mL), extracted with dichloromethane (50 mL*3), and the organic phase was dried over magnesium sulfate and concentrated to obtain a yellow oily product (5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-phenyl)methyl methanesulfonate (3.8 g, crude product).

[0444] Step F: To N,N-dimethylformamide (50 mL) in which (5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-phenyl)methyl methanesulfonate (3.7 g, crude) was dissolved, 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.38 g, 8.14 mmol, 1 eq) and potassium carbonate (2.25 g, 16.28 mmol, 2 eq) were added at room temperature and stirred at 25 °C for 16 hours. The mixture was diluted with water (50 mL), extracted with ethyl acetate (50 mL*3), and the organic layer was dried over magnesium sulfate, filtered and concentrated to give a yellow oily product 1-((5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-phenyl)methyl)-6-chloro-pyrazolo[3,4-d]pyrimidin-4-amine (4.3 g, crude product). ESI-MS m / z=528.2[M+H] + ;Calcu.=527.2.

[0445] Step G: At 0°C, tetrabutylammonium fluoride (1M, 8.14 mL) was added to tetrahydrofuran (70 mL) in which 1-((5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-phenyl)methyl)-6-chloro-pyrazolo[3,4-d]pyrimidin-4-amine (4.3 g, crude product) was dissolved, and the mixture was stirred at 25°C for 2 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (100 ml*3). The organic layer was dried over magnesium sulfate, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 40g Silica gel chromatography column, eluent 0-53% ethyl acetate / petroleum ether) to give a yellow solid product (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-4-fluoro-phenyl)methoxy)phenyl)methanol (930 mg, three-step yield 28%).

[0446] Step H: At 0°C, sodium hydride (58.0 mg, 1.45 mmol, 60% content, 3 eq) was added to a solution of (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-4-fluoro-phenyl)methoxy)phenyl)methanol (200 mg, 483.3 μmol, 1 eq) in N-methylpyrrolidone (3 mL), and stirred at 110°C for 2 hours. The mixture was filtered and then purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 30%-70% B in 36 minutes) to obtain compound A-31, yellow solid product 3 6 -Fluoro-1 1 H-5,8-Dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-1 4 -amine (7.75 mg, yield 4%). 1 H NMR: (400MHz, DMSO-d 6 )δppm 8.02-8.11(m,1H),7.90(s,1H),7.56-7.82(m,1H),7.43-7.56(m,2H),7.41(dt,J=2.3,5.4Hz,1H),7.00 -7.15(m,2H),6.86(d,J=7.3Hz,1H),6.65(dd,J=1.8,8.2Hz,1H),5.43(s,2H),5.38(s,2H),5.22(s,2H). ESI-MS m / z=378.1[M+H] + ;Calcu.=377.1.

[0447] Example 29: Synthesis of Compound A-32

[0448]

[0449] Step A: Sodium borohydride (12.23 g, 323.3 mmol) was added to a methanol solution (300 mL) of 5-formyl-2-methoxy-benzoic acid methyl ester (30 g, 154.5 mmol) at 0°C. The mixture was stirred at 0°C for 2 hours. Diluted with water (120 mL) at 0°C, extracted with ethyl acetate (120 mL*3), and dried over magnesium sulfate. Filtered and concentrated under reduced pressure to obtain 5-(hydroxymethyl)-2-methoxy-benzoic acid methyl ester (29 g, 96%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6)δ7.62(d,J=2.3Hz,1H),7.46(dd,J=2.3,8.5Hz,1H),7.10(d,J=8.6Hz,1H),5.19(t,J=5.8Hz,1H),4.45(d,J=5.8Hz,2H),3.81(s,3H),3.79(s,3H).

[0450] Step B: Add triethylamine (44.87 g, 443.4 mmol, 61.7 mL) to a solution of 5-(hydroxymethyl)-2-methoxy-benzoic acid methyl ester (29 g, 147.81 mmol) in dichloromethane (300 mL), add methanesulfonyl chloride (42.33 g, 369.5 mmol, 28.6 mL) at 0°C, and stir at 25°C for 2 hours. The reaction solution was quenched with water (200 mL) at 0°C. The reaction mixture was extracted with dichloromethane (100 mL*3). Dry over magnesium sulfate, filter and concentrate under reduced pressure to obtain 5-(chloromethyl)-2-methoxy-benzoic acid methyl ester (28 g, 88%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ7.73(d,J=2.4Hz,1H),7.61(dd,J=2.4,8.6Hz,1H),7.16(d,J=8.8Hz,1H),4.77(s,2H),3.83(s,3H),3.79(s,3H).

[0451] Step C: Dissolve 5-(chloromethyl)-2-methoxy-benzoic acid methyl ester (10 g, 36.46 mmol) in acetonitrile (100 mL), add cesium carbonate (35.64 g, 109.4 mmol) and 3-(((tert-butyldimethylsilyl)oxy)methyl)phenol (9.56 g, 40.1 mmol), stir at 50°C for 12 hours, filter and concentrate the mixture at 60°C, and perform flash silica gel chromatography ( 80g Silica Flash column, eluent: 0-15% ethyl acetate / petroleum ether gradient @100 mL / min). The obtained product was methyl 5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-2-methoxybenzoate (9.4 g, 62%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6)δ7.72(br s,1H),7.54-7.64(m,1H),7.20-7.27(m,1H),7.09-7.18(m,1H),6.67-6.97(m,3H),5.04(br s,2H),4.66(br s, 2H), 3.82 (d, J = 1.9Hz, 3H), 3.78 (br d, J = 2.0Hz, 3H), 0.84-0.93 (m, 9H), 0.09--0.02 (m, 6H).

[0452] Step D: To a tetrahydrofuran solution (50 mL) of methyl 5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-2-methoxybenzoate (5 g, 12.0 mmol) was added diisobutylaluminum hydride (1 M, 60 mL) at 0°C. The mixture was stirred at 0°C for 2 hours. It was quenched with ice water (200 mL), 1N hydrochloric acid was added to pH=8, and the mixture was extracted with ethyl acetate (200 mL*2). The organic layer was dried over magnesium sulfate, filtered, and concentrated. The obtained compound (5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-2-methoxyphenyl)methanol (5.1 g, crude product) was a white solid. 1 H NMR (400 MHz, DMSO-d 6 )δ7.39(d,J=1.5Hz,1H),7.16-7.22(m,2H),6.84-6.88(m,2H),6.80(dd,J=1.7,7.8Hz,2H),4.97(t,J =5.6Hz,1H),4.93(s,2H),4.60(s,2H),4.43(d,J=5.6Hz,2H),3.70(s,3H),0.83(s,9H),0.00(s,6H).

[0453] Step E: To a solution of (5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-2-methoxyphenyl)methanol (4.2 g, 10.81 mmol) in dichloromethane (50 mL) was added triethylamine (2.19 g, 21.62 mmol, 3.01 mL), and methanesulfonyl chloride (3.22 g, 28.11 mmol, 2.18 mL) was added at 0°C and stirred at 0°C for 2 hours. The reaction mixture was quenched with water (100 mL) at 0°C. The reaction mixture was extracted with dichloromethane (100 mL*3). Drying over magnesium sulfate, filtering and concentration under reduced pressure gave 5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-2-methoxybenzyl methanesulfonate (5.6 g, crude) as a yellow oil.

[0454] Step F: To a solution of 5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-2-methoxybenzyl methanesulfonate (5.6 g, crude) in N,N-dimethylformamide (50 mL) were added potassium carbonate (4.98 g, 36.0 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.93 g, 11.4 mmol) and stirred at 50° C. for 12 hours. The mixture was filtered and concentrated to give 1-(5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-2-methoxybenzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (6 g, crude) as a white solid. ESI-MS m / z=540.2[M+H] + ;Calcu.=539.2.

[0455] Step G: To a solution of 1-(5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-2-methoxybenzyl)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (6 g, crude) in tetrahydrofuran (60 mL), tetrabutylammonium fluoride (1 M, 16.7 mL) was added and stirred at 25°C for 2 hours. The mixture was filtered and the mother liquor was concentrated. The residue was purified by flash silica gel chromatography ( 80g Silica Flash column, eluent: 0-34% tetrahydrofuran / petroleum ether gradient @100 mL / min). A white solid compound (3-((3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-4-methoxybenzyl)oxy)phenyl)methanol (252 mg, four-step yield 6%) was obtained. ESI-MS m / z=426.1[M+H] + ;Calcu.=425.1.

[0456] Step H: To a solution of (3-((3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-4-methoxybenzyl)oxy)phenyl)methanol (100 mg, 234.8 μmol) in N-methylpyrrolidone (2 mL) was added sodium hydride (28.2 mg, 704.4 μmol, 60% purity). The mixture was stirred at 110°C for 12 hours. The reaction mixture was extracted with ethyl acetate (10 mL*2) and water (10 mL). The combined organic layers were washed with brine (50 mL*2), dried over magnesium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. It was then purified by preparative high performance liquid chromatography (hydrochloric acid conditions) to obtain compound A-32, a white solid 3 6 -Methoxy-1 1H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (6.68 mg, 7%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.46(br s,1H),7.89(s,1H),7.38-7.71(m,4H),7.30-7.36(m,1H),7.09(t,J=7.9Hz,1H),6.92(d,J=8.5Hz,1H),6.84(br d,J=7.3Hz,1H),6.60-6.70(m,1H),5.34(s,4H),5.10(s,2H),3.69(s,3H). ESI-MS m / z=390.2[M+H] + ;Calcu.=389.2.

[0457] Example 30: Synthesis of Compound A-33 / M-8 / M-9

[0458]

[0459] Step A: Sodium borohydride (2.78 g, 73.49 mmol, 1.79 eq) was added to methanol (150 mL) at 0°C in which methyl 2-bromo-5-formylbenzoate (10 g, 41.14 mmol, 1 eq) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. Water (100 mL) was added to the reaction solution and extracted with ethyl acetate (200 mL*3) for 3 times. The organic phase was dried over magnesium sulfate, filtered and concentrated to obtain a colorless oily product, methyl 2-bromo-5-(hydroxymethyl)benzoate (6.8 g, yield 67%). ESI-MS m / z=244.8[M+H] + ;Calcu.=244.0

[0460] Step B: At 0°C, triethylamine (8.42 g, 83.24 mmol, 11.59 mL, 3 eq) and methanesulfonyl chloride (4.8 g, 41.90 mmol, 3.24 mL, 1.51 eq) were added to dichloromethane (150 mL) in which methyl 2-bromo-5-(hydroxymethyl)benzoate (6.8 g, 27.75 mmol, 1 eq) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 hours. The reaction solution was poured into water (150 mL), extracted with dichloromethane (100 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product, methyl 2-bromo-5-(chloromethyl)benzoate (5.36 g, yield 73%). 1 H NMR: (400MHz, DMSO-d 6)δppm 7.85(d,J=2.1Hz,1H),7.78(d,J=8.3Hz,1H),7.55(dd,J=2.3,8.3Hz,1H),4.81(s,2H),3.87(s,3H).

[0461] Step C: At room temperature, 3-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (4.76 g, 19.96 mmol, 1 eq) and cesium carbonate (19.51 g, 59.88 mmol, 3 eq) were added to acetonitrile (80 mL) in which methyl 2-bromo-5-(chloromethyl)benzoate (5.26 g, 19.96 mmol, 1 eq) was dissolved, and the mixture was stirred at 50°C for 16 hours. The reaction solution was poured into water (100 mL), extracted with ethyl acetate (150 mL*3), the organic phase was dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product, methyl 2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzoate (6.2 g, yield 67%). 1 HNMR: (400MHz, DMSO-d 6 )δppm 7.78(d,J=1.8Hz,1H),7.72(d,J=8.3Hz,1H),7.48(dd,J=1.8,8.2Hz,1H),7.21(t,J=7.8Hz ,1H),6.91-6.80(m,3H),5.09(s,2H),4.62(s,2H),3.81(s,3H),0.83(s,9H),0.00(s,6H).

[0462] Step D: Diisobutylaluminum hydride (1M, 64.5mL, 5eq) was added to tetrahydrofuran (60mL) in which methyl 2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzoate (6g, 12.89mmol, 1eq) was dissolved at 0°C. The mixture was stirred at 0°C for 2 hours. The mixture was quenched with ice water (50mL) and dilute hydrochloric acid (1N, 10mL), extracted with ethyl acetate (50mL*3), and the organic layer was dried over magnesium sulfate, filtered and concentrated to give a yellow oily product (2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (5.6g, yield 99%). 1 HNMR: (400MHz, DMSO-d 6)δppm 7.56(d,J=1.9Hz,1H),7.51(d,J=8.1Hz,1H),7.15-7.24(m,3H),6.86(d,J=1.8Hz,1H),6.82(s,1H) ,5.43(t,J=5.5Hz,1H),5.04(s,2H),4.61(s,2H),4.46(d,J=5.5Hz,2H),0.83(s,9H),0.00(s,6H).

[0463] Step E: At 0°C, triethylamine (3.88 g, 38.34 mmol, 5.34 mL) and methanesulfonyl chloride (2.67 g, 23.31 mmol, 1.80 mL) were added to dichloromethane (100 mL) in which (2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (5.59 g, 12.78 mmol) was dissolved. After the addition, the whole mixture was stirred at 25°C for 2 h. The reaction solution was poured into water (100 mL), extracted with dichloromethane (100 mL*3), and the organic phase was dried and concentrated over magnesium sulfate to obtain a yellow oily product (2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl methanesulfonate (7.09 g, crude product). 1 H NMR: (400MHz, DMSO-d 6 )δppm 7.72-7.55(m,2H),7.37(dd,J=2.1,8.2Hz,1H),7.24-7.16(m,1H),6.93-6.77(m,3H),5. 30-5.10(m,2H),5.06(s,2H),4.61(s,2H),3.22(s,3H),0.83(s,9H),0.04--0.01(m,6H).

[0464] Step F: To N,N-dimethylformamide (120 mL) in which (2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl methanesulfonate (6.99 g, crude) was dissolved, 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2.30 g, 13.56 mmol) and potassium carbonate (3.75 g, 27.13 mmol) were added at room temperature. l), stirred at 25 ° C for 16 hours. The mixture was diluted with water (50 mL), extracted with ethyl acetate (50 mL * 3), the organic layer was dried over magnesium sulfate, filtered and concentrated to obtain a yellow oily product 1-((2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl)-6-chloro-pyrazolo[3,4-d]pyrimidin-4-amine (8.47 g, crude product). ESI-MS m / z=590.1[M+H] + ;Calcu.=589.1.

[0465] Step G: At 0°C, tetrabutylammonium fluoride (1M, 21.3 mL) was added to tetrahydrofuran (140 mL) in which 1-((2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl)-6-chloro-pyrazolo[3,4-d]pyrimidin-4-amine (8.37 g, crude product) was dissolved, and the mixture was stirred at 25°C for 2 hours. The mixture was diluted with 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 Silica gel chromatography column, eluent 0-52% ethyl acetate / petroleum ether) to obtain a yellow solid product (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-4-bromo-phenyl)methoxy)phenyl)methanol (1.2 g, three-step yield 20%). ESI-MS m / z=476.0[M+H] + ;Calcu.=475.0.

[0466] Step H: At 0°C, sodium hydride (278 mg, 6.95 mmol, 60% content, 3 eq) was added to a solution of (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-4-bromo-phenyl)methoxy)phenyl)methanol (1.1 g, 2.32 mmol) dissolved in N-methylpyrrolidone (20 mL), and stirred at 110°C for 2 hours. The mixture was diluted with water (20 mL) and extracted with tetrahydrofuran / ethyl acetate (1 / 1, 20 mL*3). The organic layer was dried over magnesium sulfate, filtered and concentrated to obtain M-8, a yellow solid product 3 6 -Bromo-11 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (0.6 g, yield 35%). ESI-MS m / z = 439.8 [M+H] + ;Calcu.=439.1.

[0467] Step I: Dissolve 3 6 -Bromo-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (300 mg, 684.5 μmol) in methanol (5 mL) and N, N-dimethylformamide (5 mL) were added with triethylamine (138 mg, 1.37 mmol, 190 μL) and 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (50.1 mg, 68.45 μmol), and stirred for 4 days under carbon monoxide (1 MPa pressure) and 100 ° C. The mixture was concentrated to obtain M-9, a yellow solid methyl 1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-3 6 -carboxylate (477 mg, crude product). ESI-MS m / z=418.1[M+H] + ;Calcu.=417.1.

[0468] Step J: At room temperature, dissolve methyl 1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-3 6 -carboxylate (50 mg, crude product) in tetrahydrofuran (1 mL) and water (1 mL), sodium hydroxide (19.2 mg, 479.1 μmol) was added and stirred at 60 ° C for 2 hours. The mixture was diluted with water (2 mL), extracted with ethyl acetate (2 mL * 3), the organic layer was dried over magnesium sulfate, filtered and concentrated. Then it was purified by preparative high performance liquid chromatography (chromatographic column: XtimateC18 150 * 40mm * 10um; mobile phase: [water (formic acid) - methanol]; gradient: 26% -66% B in 36 minutes) to obtain compound A-33, yellow solid product 1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-3 6-carboxylic acid (4.62 mg, two-step yield 16%). 1 H NMR: (400MHz, DMSO-d 6 )δppm 7.98(s,1H),7.47-7.91(m,3H),7.38(br d,J=7.9Hz,1H),7.19-7.33(m,2H),7.12(t,J=7.9Hz,1H),6.85(d,J=7.4H z, 1H), 6.76 (dd, J = 1.8, 8.3Hz, 1H), 5.82 (s, 2H), 5.32 (s, 2H), 5.07 (s, 2H). ESI-MS m / z=404.1[M+H] + ;Calcu.=403.1.

[0469] Example 31: Synthesis of Compound A-34

[0470]

[0471] Step A: At 0°C, dissolve methyl 1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-3 6 -carboxylate (100 mg, 239.57 μmol) in tetrahydrofuran (2 mL) was added with lithium aluminum tetrahydride (2.5 M, 191.7 μL) and stirred at 0°C for 2 hours. The mixture was quenched with water (0.2 mL), 15% sodium hydroxide solution (0.2 mL) and water (0.4 mL), and then filtered. It was then purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 22%-62% B in 36 minutes) to obtain compound A-34, a white solid product (1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-3 6 -yl)methanol (5.45 mg, yield 6%). 1 H NMR: (400MHz, DMSO-d 6)δppm 7.93(s,1H),7.87(s,1H),7.58(m,3H),7.34-7.41(m,2H),7.02-7.15(m,1H),6.85(d,J= 7.5Hz,1H),6.64(dd,J=1.9,8.3Hz,1H),5.44(s,2H),5.37(s,2H),5.20(s,2H),4.65(br d,J=5.0Hz,2H). ESI-MS m / z=390.1[M+H] + ;Calcu.=389.2.

[0472] Example 32: Synthesis of Compound A-35 / M-10

[0473]

[0474] Step A: Potassium carbonate (15.34 g, 111.0 mmol) was added to acetonitrile (200 mL) dissolved with 3-formyl-4-hydroxy-benzoic acid methyl ester (20 g, 111.0 mmol) at room temperature, and the whole mixture was stirred at 50°C for 16 hours. The reaction solution was filtered and concentrated to obtain a light yellow oily product to obtain 4-allyloxy-3-formylbenzoic acid methyl ester (18 g, 74%). 1 H NMR (400 MHz, DMSO-d 6 )δ10.37(s,1H),8.23(d,J=2.3Hz,1H),8.14-8.19(m,1H),7.34(d,J=8.8Hz,1H),6.07-6.16 (m,1H),5.41-5.55(m,1H),5.33(dd,J=1.4,10.6Hz,1H),4.82(d,J=5.0Hz,2H),3.84(s,3H).

[0475] Step B: At room temperature, sodium borohydride (6.16 g, 162.82 mmol) was added to methanol (180 mL) in which methyl 4-allyloxy-3-formylbenzoate (18 g, 81.74 mmol) was dissolved. After the addition, the whole mixture was stirred at 0°C for 2 hours. The reaction solution was poured into water (100 mL), extracted with ethyl acetate (120 mL*3), and the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was separated on a silica gel column (silica, 0-9% petroleum ether: ethyl acetate, @100 mL / min) to obtain a yellow oily product, methyl 4-allyloxy-3-(hydroxymethyl)benzoate (16 g, 88%). 1 H NMR (400 MHz, DMSO-d 6)δ8.04(d,J=1.5Hz,1H),7.84(dd,J=2.1,8.6Hz,1H),7.02-7.09(m,1H),5.97-6.14(m,1H) ,5.36-5.45(m,1H),5.19-5.30(m,2H),4.64-4.71(m,2H),4.51-4.56(m,2H),3.82(s,3H). ESI-MS m / z=205.1[M-OH] + ;Calcu.=222.1.

[0476] Step C: Add triethylamine (20.49 g, 202.49 mmol, 28.18 mL) to a solution of methyl 4-allyloxy-3-(hydroxymethyl)benzoate (15 g, 67.50 mmol) in dichloromethane (150 mL), add methanesulfonyl chloride (15.55 g, 135.75 mmol, 10.51 mL) dropwise at 0°C, and stir at 0°C for 2 hours. Pour the reaction solution into water (120 mL), extract with dichloromethane (120 mL*3), dry the organic phase with magnesium sulfate, filter and concentrate. Obtain a yellow oily product, methyl 4-allyloxy-3-(methylsulfonyloxymethyl)benzoate (16.2 g, crude product).

[0477] Step D: Potassium carbonate (11.04 g, 79.91 mmol) and 3-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (6.67 g, 27.97 mmol) were added to a solution of methyl 4-allyloxy-3-(methylsulfonyloxymethyl)benzoate (8 g, crude product) in acetonitrile (80 mL), and the mixture was stirred at 40° C. for 16 hours. The mixture was filtered and concentrated, and the residue was purified by flash silica gel chromatography ( 40g fast The mixture was filtered through a silica gel column with 0-15% tetrahydrofuran / petroleum ether as the eluent to give a yellow oily product, methyl 4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzoate (8.5 g, crude product).

[0478] Step E: Compound 4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzoic acid methyl ester (8.5 g, crude product) was dissolved in tetrahydrofuran (90 mL), and then diisobutylaluminum hydride (1M, 96 mL) was added dropwise at 0°C and stirred for 2 hours at 0°C. The mixture was poured into water (120 mL) and extracted with ethyl acetate (120 mL*3). The organic phase was dried over magnesium sulfate, filtered and concentrated, and flash silica gel chromatography ( 40g fast Silica gel chromatography column, eluent 0-8% ethyl acetate / petroleum ether) to obtain a yellow oily product (4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (4.5 g, three-step yield 33%). 1 H NMR (400 MHz, DMSO-d 6 )δ7.29(d,J=1.1Hz,1H),7.14-7.21(m,2H),6.93(d,J=8.4Hz,1H),6.88(s,1H),6.81(d,J=6.6Hz,2H),5.86-6.02(m,1H),5.32(dd,J=1.6,1 7.3Hz, 1H), 5.16 (d, J = 10.5Hz, 1H), 4.95-5.06 (m, 3H), 4.62 (s, 2H), 4.55 (d, J = 4.8Hz, 2H), 4.35 (d, J = 5.6Hz, 2H), 0.83 (s, 9H), 0.00 (s, 6H). ESI-MS m / z=497.2[M-OH] + ;Calcu.=414.2.

[0479] Step F: Dissolve the compound (4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (4.5 g, 10.85 mmol) in dichloromethane (50 mL), add triethylamine (3.29 g, 32.56 mmol, 4.5 mL), add methanesulfonyl chloride (3.12 g, 27.24 mmol, 2.1 mL) at 0°C, and stir at 25°C for 3 hours. The mixture was poured into water (120 mL), extracted with ethyl acetate (120 mL*3), and the organic phase was dried over magnesium sulfate, filtered, and concentrated. The yellow oily product (4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl methanesulfonate (4.7 g, crude product) was obtained.

[0480] Step G: To a solution of compound (4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl methanesulfonate (4.7 g, crude) in N,N-dimethylformamide (50 mL), potassium carbonate (3.96 g, 28.62 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.62 g, 9.54 mmol) were added and stirred at 50°C for 16 hours. The mixture was filtered and concentrated, and the residue was purified by flash silica gel chromatography ( 120g Silica gel chromatography column, eluent 0-32% tetrahydrofuran / petroleum ether gradient @100 mL / min) to give a yellow solid product 1-((4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl)-6-chloropyrazolo[3,4-d]pyrimidin-4-amine (2.1 g, 3.71 mmol, two-step yield 35%). 1 H NMR (400 MHz, DMSO-d 6 )δ8.22(br s,1H),8.10(br s,1H),8.03(s,1H),7.27(d,J=2.0Hz,1H),7.07-7.21(m,2H),6.94(d,J=8.5Hz,1H),6.76-6.86(m,3H),5.82-6.07(m,1H) ,5.27-5.35(m,3H),5.16(dd,J=1.6,10.6Hz,1H),4.97(s,2H),4.61(s,2H),4.51-4.57(m,2H),0.83(s,9H),0.00(s,6H). ESI-MS m / z=566.2[M+H] + ;Calcu.=565.2.

[0481] Step H: Tetrabutylammonium fluoride (1M, 5.2 mL) was added to a solution of the compound 1-((4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl)-6-chloro-pyrazolo[3,4-d]pyrimidin-4-amine (2.1 g, 3.71 mmol) in tetrahydrofuran (20 mL), and the mixture was stirred at 25°C for 5 hours. Then, the mixture was diluted with 20 mL of water, extracted with ethyl acetate (20 mL*3), the organic layer was dried over magnesium sulfate, filtered and concentrated to give a yellow solid product (3-((2-allyloxy-5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)methoxy)phenyl)methanol (1.55 g, 92%). ESI-MS m / z=352.2[M+H] + ;Calcu.=451.1.

[0482] Step I: Sodium hydride (398.31 mg, 9.96 mmol, content 60%) was added to the compound (3-((2-allyloxy-5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)methoxy)phenyl)methanol (1.5 g, 3.32 mmol) in N-methylpyrrolidone (15 mL), and the mixture was stirred at 80°C for 10 hours. Then diluted with water (15 mL), extracted with ethyl acetate (15 mL*3), the organic layer was dried over magnesium sulfate, filtered and concentrated, and flash silica gel chromatography ( 120g fast Purification by silica gel chromatography (eluting with 0-50% tetrahydrofuran / petroleum ether gradient @100 mL / min) gave M-10 as a yellow solid product 3 4 -(allyloxy)-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (300 mg, 22%). ESI-MS m / z=416.2[M+H] + ;Calcu.=415.2.

[0483] Step J: To compound 3 4 -(allyloxy)-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (100 mg, 240.7 μmol) in dichloromethane (10.5 mL) solution was added with tetrakis(triphenylphosphine)palladium (139 mg, 120.3 μmol) and acetic acid (14.5 mg, 240.7 μmol, 13.8 μL), and tributyltin hydride (350 mg, 1.20 mmol, 319 μL) was added at 0°C. Stir at 25°C for 2 hours. The mixture was filtered and concentrated, filtered and concentrated, and then purified by reverse phase high performance liquid chromatography (chromatographic column: C18 150*40mm*10um, mobile phase: [water (formic acid)-methanol], gradient: 24%~64% B, 25min) to obtain compound A-35, white solid product 1 4 -Amino-1 1 H-5,8-dioxa-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzylcyclooctane-3 4 -alcohol (7.1 mg, 8%). 1 H NMR (400 MHz, DMSO-d 6)δ9.64(s,1H),7.88(d,J=2.0Hz,1H),7.86(s,1H),7.58(m,3H),7.05-7.11(m,1H),7.02(dd,J=2.1,8. 3Hz, 1H), 6.86 (d, J = 7.5Hz, 1H), 6.68 (s, 1H), 6.64-6.66 (m, 1H), 5.39 (s, 2H), 5.24 (s, 2H), 5.20 (s, 2H). ESI-MS m / z=376.3[M+H] + ;Calcu.=375.1.

[0484] Test Example 1: Evaluation of drug efficacy at the cellular level

[0485] Purpose: Using HEK-Blue TM hTLR7 in vitro cell model to evaluate the TLR7 agonist activity of compounds

[0486] Experimental Materials:

[0487] 1 Cell line: HEK-Blue TM hTLR7 (InvivoGen)

[0488] 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

[0489] (GIBCO)

[0490] 3 Instruments: Microplate reader (PerkinElmer, Multimode plate reader 2014), CO 2 Incubator (THERMO, STERI-CYCLEI160), cell counter (BECKMANCOULTER, vi-cell XR)

[0491] Experimental methods:

[0492] Step 1: Cell plating

[0493] a) Discard the culture medium in the T150 flask, add 10 mL of pre-warmed DPBS, and gently shake the flask to rinse the cells. After discarding the DPBS, add 8.0 mL of TrypLETM Add the Express enzyme to the T150 culture flask and gently shake the flask to allow it to infiltrate all cells.

[0494] b) Add TrypLE TM Express enzyme was discarded and the tube was placed in an incubator for 1 to 2 minutes.

[0495] c) Add serum-containing culture medium to terminate digestion, collect the cell suspension, centrifuge at 300r at room temperature for 5 minutes, and discard the supernatant.

[0496] d) Add fresh culture medium, mix the cells by pipetting, and then count the cells using a vi-cell XR cell counter.

[0497] e) Dilute the cells to a density of 6.25*10 with fresh culture medium. 5 cells / mL, 80 μL of the diluted cell suspension was inoculated into a 96-well plate, i.e., 5*10 4 cells / well.

[0498] f) Place the 96-well plate in an incubator and incubate overnight (5% CO 2 , 37°C).

[0499] Step 2: Drug treatment

[0500] a) The test compound was prepared into a 10 mM stock solution in DMSO;

[0501] b) The compound to be tested was diluted from 10 mM to 0.1 mM using culture medium, and then the compound was diluted 3-fold using culture medium containing 1% DMSO, for a total of 8 concentration gradients, with DMSO as a negative control, and R848 and GS9620 as positive controls.

[0502] c) Take 20 μL of the diluted compound and add it to a 96-well plate inoculated with 80 μL of cells, dilute the drug 5 times, and the final concentration of DMSO in each well is 0.2%. The final concentrations of the test compound and the control compound are 20000, 6667, 2222, 741, 247, 82, 27, and 9 nM.

[0503] d) Place the 96-well plate in a 5% CO 2 , incubate in a 37°C incubator for 24 hours.

[0504] Step 3: Activity Detection

[0505] a) Take 10 μL of the cell supernatant after compound incubation and transfer it to a 96-well transparent test plate, then add 90 μL of QUANTI-Blue to each well of the test plate. TM Solution, 5% CO 2, incubate in a 37°C incubator for 1 hour.

[0506] b) The light absorption of secretory alkaline phosphatase (SEAP) at 650 nm was detected using an ELISA reader.

[0507] Data Analysis:

[0508] The activation factor was calculated using the following formula: activation factor = OD 650 Treatment group / OD 650 The GraphPadPrism 7 software was used to fit the curve according to the concentration of the compound and the corresponding activation factor, and the EC 50 value.

[0509] Reference substances: R848 (InvivoGen) and GS-9620 (CAYMAN), the structures are shown below:

[0510]

[0511] The experimental results are shown in the following table:

[0512]

[0513]

[0514] Test Example 2: Test of the agonistic activity of the compounds of the present invention on human peripheral blood mononuclear cells

[0515] Human peripheral blood mononuclear cells (hPBMCs) represent the main human immune cells in the blood, including lymphocytes, monocytes and dendritic cells. These cells express TLR7, TLR8 or TLR9 and respond to stimulation by the corresponding ligands. In this experiment, hPBMCs were stimulated with potential TLR7 agonists, and the levels of IFN-α, TNF-α and IL-12p40 were detected to reflect the activation of the compounds on the TLR7 receptor.

[0516] Experimental Materials:

[0517] 1. Cell line: hPBMCs (Sailybio), culture conditions: RPMI 1640 medium + 10% (v / v) FBS + 1% double antibody (Penicillin-Streptomycin), 5% CO 2 , 37℃.

[0518] 2. Reagents: RPMI 1640 medium (GIBCO), FBS (GIBCO), Penicillin-Streptomycin (GIBCO), Human IL-12 / IL-23p40 DuoSet ELISA (R&D), Human IFN-alpha 2 / IFNA2 DuoSet ELISA (R&D), Human TNF-alpha ELISA (R&D).

[0519] 3. Instrument: Microplate reader (perkinElmer, Multimode plate reader 2014), CO 2 Incubator (THERMO, STERI-CYCLEI160), cell counter (BECKMAN COULTER, vi-cell XR).

[0520] Experimental methods:

[0521] a) Cryopreserved hPBMCs were thawed in a 37°C water bath and transferred to 1640 complete medium. After centrifugation at 500 r / min at room temperature for 10 minutes, the supernatant was discarded. The cells were resuspended in 1640 complete medium and transferred to a culture flask. Incubate at 37°C and 5% CO 2 Incubate overnight under the same conditions.

[0522] b) Collect hPBMCs after stable culture, count and dilute the cells to 1.11*10 7 cells / mL, 180 μL of the diluted cell suspension was inoculated into a 96-well plate, i.e. 2*10 6 The 96-well plate was placed in a constant temperature incubator and incubated for 24 h (5% CO 2 , 37°C).

[0523] c) The test compound was prepared into a 10mM stock solution with DMSO and diluted to 0.03mM (IFN-α detection) and 0.1mM (TNF-α, IL-12p40 detection), respectively. Then, the compound was diluted into 8 concentration gradients with culture medium containing 2% DMSO. The final maximum concentration of the compound was 30uM (IFN-α detection) and 100uM (TNF-α, IL-12p40 detection), respectively.

[0524] 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%.

[0525] e) After incubation at 37° C. for 24 hours, the cell culture supernatant was collected by centrifugation, and the levels of IFN-α, TNF-α, and IL-12p40 in the supernatant were determined using an ELISA detection kit.

[0526] f) The light absorption at 450 nm was measured by an enzyme-labeled instrument, and the level of secretion of the corresponding cytokines was quantitatively determined according to the corresponding standard curve, and the EC was calculated by fitting the curve. 50 The experimental results are shown in the following table:

[0527]

Claims

1. A compound having a structure as shown in formula (I), a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or a metabolite thereof, in, R 1 Selected from: hydrogen, halogen, hydroxyl, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl; Q is selected from -O-, -S-, -NH-; Ring A is selected from phenyl, 5-6 membered heteroaryl, naphthyl ring and bicyclic heteroaryl, wherein the phenyl, 5-6 membered heteroaryl, naphthyl ring 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; 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 phenyl and naphthalene, and the phenyl and naphthalene rings 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’ 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: 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-, -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; 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)-, -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, and p and r cannot be 0 at the same time, 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 any one of claims 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 selected from bicyclic heteroaryl substituted by one or more substituents, for example 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, 1,2,3,4-tetrahydroquinoxaline; Ring B is selected from a naphthalene ring.

5. 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 selected from a benzene ring, a 5-6 membered heteroaryl group, and optionally, Ring A is substituted by 1, 2 or 3 substituents; Wherein, the substituents of ring A are each independently 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 2 R 3 、-CH2R a 、-C(=O)R b 、-OR c , -SH, -(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 2 , R 3 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: halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy.

6. 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 B is selected from a benzene ring, and optionally, ring B is substituted by 1, 2 or 3 substituents; The substituents of ring B are each independently 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’ 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: halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy.

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: The substituents of Ring A and Ring B are each independently selected from the following groups: C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, phenyl, 5-6 membered heteroaryl.

8. 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: The substituents of ring A are selected from the following groups: hydrogen, halogen, nitro, 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 ; Preferably, 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, -N(C1-C6 alkyl)2; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more C1-C6 alkyl groups; Preferably, R 3 , R 4 are each independently selected from hydrogen, C 1-6 alkyl; The substituents of ring B are selected from the following groups: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)R b’ 、-OR c’ ; Preferably, R a’ , R b’ , R c’ 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 C1-C6 alkyl groups; Preferably, R 3’ , R 4’ are each independently selected from hydrogen, C 1-6 alkyl.

9. The compound of 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: Ring A is selected from phenyl, 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are optionally substituted by 1, 2 or 3 groups selected from the following: halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxyl, C1-C6 alkoxy, =O, -C(=O)H, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, -OCH2CH=CH2; 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; Preferably, the 5-6 membered heteroaryl is a monocyclic heteroaryl, such as a 5-6 membered monocyclic heteroaryl containing nitrogen, oxygen and / or sulfur; Preferably, the 5-6 membered heteroaryl group is selected from the following groups formed by heteroaromatic rings:

10. The compound of 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: Ring B is phenyl, and the phenyl is optionally substituted by 1, 2 or 3 groups selected from the following: halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxyl, C1-C6 alkoxy, =O, -C(=O)H, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -NR 3’ R 4’ , carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, -OCH2CH=CH2; Among them, 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.

11. The compound of 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: L is selected from -(CH2) m -W-(CH2) n -, wherein W 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)-, -S-, -S(=O)-, -S(=O)(=O)-, -P(=O)(OH)-, -OC(=O)-, -C(=O)O-, -NHS(=O)(=O)-, -S(=O)(=O)NH-, and m and n are each independently selected from 0, 1 or 2; Preferably, L is selected from the following structures: -O-CH2-, -NH-CH2-, -CH2-, -NH-, -NH-C(=O)-, -CH2-O-, -CH2-O-CH2-, -CH2-CH2-, -O-CH2-CH2-; More preferably, L is selected from the following structures: -O-CH2-, -CH2-O-, -NH-CH2-, -CH2-NH-, -CH2-CH2-.

12. The compound of 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: q is 0.

13. The compound of any one of claims 1 to 12, 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 is selected from -CH2-.

14. The compound of any one of claims 1 to 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 (I'): Among them, R 1 , Q, L, Ring A, Ring B are as defined in any one of 1-9; R 2 Selected from hydrogen or C1-C6 alkyl.

15. The compound of any one of claims 1 to 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 (II): Among them, R 1 , Q, L are as defined in any one of claims 1-3 and 10 respectively; R 2 is hydrogen or C1-C6 alkyl; R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, =O, -C(=O)H, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, -OCH2CH=CH2; 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, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy.

16. The compound of any one of claims 1 to 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): Among them, R 1 , Q, L are as defined in any one of claims 1-3 and 10 respectively; R 2 is hydrogen or C1-C6 alkyl; R 5 , R 6 Each is independently selected from: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, =O, -C(=O)H, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, -OCH2CH=CH2; 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, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy.

17. The compound of any one of claims 1 to 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 (IV): Among them, R 1 , Q, L are as defined in any one of claims 1-3 and 10 respectively; R 2 is hydrogen or C1-C6 alkyl; One of X and Y is C or CH, and the other is N or NH; R 5 , R 7 Each is independently selected from: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, =O, -C(=O)H, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, -OCH2CH=CH2; 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, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy.

18. The compound of any one of claims 1 to 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 (VII): Among them, R 1 , Q, L are as defined in any one of claims 1, 2, 3, and 10 respectively; R 2 Selected from hydrogen or C1-C6 alkyl; R 5 , R 6 , R 7 Each is independently selected from: hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, =O, -C(=O)H, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -NR 3 R 4 , carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, -OCH2CH=CH2; 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: halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy.

19. A compound according to any one of claims 1 to 18, 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).

20. The compound of any one of claims 1-18, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystalline form, isotope-labeled substance or metabolite form thereof, wherein the compound has a structure selected from the following:

21. A pharmaceutical composition comprising a compound according to any one of claims 1 to 20, 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.

22. Use of the compound according to any one of claims 1 to 20, 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.

23. Use of the compound according to any one of claims 1 to 20, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof as a vaccine adjuvant.

24. Use of the compound according to any one of claims 1 to 20, its pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite form thereof as a photodynamic therapy agent.

25. Use of the compound according to any one of claims 1 to 20, 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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