Polyaryl-containing macrocyclic compounds and uses thereof

Abstract

The present application relates to the field of biological medicine, and particularly relates to small-molecule polyaryl-containing macrocyclic compounds which have better immunoregulation activity. Further provided in the present invention are the use of the small-molecule polyaryl-containing macrocyclic compounds in preventing or treating TLR7-related diseases, and the uses of same as vaccine adjuvants, photodynamic therapeutic agents and drug conjugates.

Description

Macrocyclic compounds containing polyaryl groups and uses thereof

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application is based on the application with CN application number 202310771533.9 and application date June 27, 2023, and claims its priority. The content of the CN application is hereby introduced as a whole into this application. Technical Field

[0003] The present application relates to the field of biomedicine, and in particular to a macrocyclic TLR7 compound containing multiple aromatic groups and its use. Background Art

[0004] Toll-like receptors (TLRs) are pattern recognition receptors expressed on the surface or inside immune cells such as dendritic cells, macrophages, and natural killer cells. Their main function is to activate immune responses and resist infection by recognizing pathogen-associated molecular patterns (PAMPs). Humans have a total of 10 TLRs, most of which 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 endosomes.

[0005] TLR7, a member of the TLR family, is primarily expressed on the endosomal membranes of plasmacytoid dendritic cells and can recognize viral single-stranded RNA (ssRNA), such as that of HIV and hepatitis C virus. TLR7 can activate the NF-κB and mitogen-activated protein kinase (MAPK) signaling pathways and promote the production of inflammatory cytokines, chemokines, and interferon kinases. The natural ligand for TLR7 is guanosine- and / or uridine-rich ssRNA, and some synthetic heterocyclic small molecules may also modulate TLR7 activity. TLR7 has three distinct domains. The N-terminal extracellular domain contains 26 leucine-rich repeats (LRRs), which form a large parallel β-sheet, allowing protein-protein interactions between the two monomers. The unstructured Z-loop region between LRR14 and LRR15 is crucial for TLR7 dimerization.

[0006] 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 have shown good anti-tumor effects. In addition, due to 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.

[0007] 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.

[0008] Summary of the Invention

[0009] The present invention aims to provide a novel, small-molecule, polyaryl-containing macrocyclic pyrazolopyrimidine TLR7 agonist with enhanced immunomodulatory activity. The present invention also provides uses of the polyaryl-containing macrocyclic pyrazolopyrimidine TLR7 agonist for preventing or treating TLR7-related diseases, as well as for use as a vaccine adjuvant and photodynamic therapy agent. The following inventions are provided:

[0010] Compound

[0011] In one aspect, the present application provides a compound having a structure as shown in formula (I), a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope label or metabolite thereof,

[0012] in,

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

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

[0015] Ring A is selected from phenyl, 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, 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 group consisting of halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, and C1-C6 alkoxy;

[0016] Ring B is selected from 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, 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 group consisting of halogen, nitro, cyano, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, and C1-C6 alkoxy;

[0017] 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 (e.g., 0, 1, 2, 3, 4, 5 or 6);

[0018] T is selected from -(CH-T 1 ) p -(Z) q -(CH-T 2 ) r-, wherein Z is selected from -O-, -NH-, -NHC(=O)-, -CH2-, -CH(C1-C6 alkyl)-, -C(C1-C6 alkyl)(C1-C6 alkyl)-, -C(=O)-, -C(=O)NH-, -N(C1-C6 alkyl)-, -N(C1-C6 alkyl)C(=O)-, -C(=O)N(C1-C6 alkyl)-, -S-, -S(=O)-, -S(=O)(=O)-, -P(=O)(C1-C6 alkyl) -C1-C6 alkyl)-, -P(=O)(C1-C6 alkoxy)-, -P(=O)(OH)-, -OC(=O)-, -C(=O)O-, -NHS(=O)(=O)-, -S(=O)(=O)NH-, -N(C1-C6 alkyl)S(=O)(=O)-, -S(=O)(=O)N(C1-C6 alkyl)-, p, q, r are each independently selected from an integer between 0 and 6 (for example, 0, 1, 2, 3, 4, 5 or 6), and p and r cannot be 0 at the same time, T 1 、T 2 Each is independently selected from: hydrogen, halogen, nitro, hydroxy, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, mercapto, -S-(C1-C6 alkyl).

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

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

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

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

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

[0024] In certain embodiments, Ring A is 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-chromene, 1,4-benzodioxane, benzomorpholine, 1,2,3,4-tetrahydroquinoxaline.

[0025] 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.

[0026] In certain embodiments, the substituents of ring A are selected from the group consisting of hydrogen, halogen, nitro, nitroso, cyano, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo-C1-C6 alkyl, =O, -NR 3 R 4 、-CH2R a 、-C(=O)R b 、-OR c , -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 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.

[0027] In certain embodiments, the substituents of Ring A are each independently selected from the following groups: C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclyl, phenyl, 5-6 membered heteroaryl.

[0028] In certain embodiments, the substituents of ring A are each independently selected from the group consisting of hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo-C1-C6 alkyl, =O, -NR 3 R 4 、-CH2R a 、-C(=O)R b 、-OR c .

[0029] In certain embodiments, 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, -N(C1-C6 alkyl)2; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more C1-C6 alkyl groups.

[0030] In certain embodiments, R 3 、R 4 are each independently selected from hydrogen, C 1-6 alkyl.

[0031] In certain embodiments, the 3-6 membered heterocyclyl is substituted by one or more groups selected from the group consisting of halogen, nitro, cyano, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, and C1-C6 alkoxy.

[0032] 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, C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl, C1-C6 alkoxy, =O, -C(=O)H, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclyl), -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.

[0033] In certain embodiments, the 5-6 membered heteroaryl is a monocyclic heteroaryl, for example, a 5-6 membered monocyclic heteroaryl containing nitrogen, oxygen and / or sulfur.

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

[0035] In certain embodiments, Ring A is selected from an optionally substituted phenyl group or an optionally substituted 5-6 membered nitrogen-containing heteroaryl group. Optionally, the 5-6 membered nitrogen-containing heteroaryl group further contains 1-2 oxygen atoms or sulfur atoms. The 5-6 membered nitrogen-containing heteroaryl group may contain 1, 2, 3 or 4 nitrogen atoms. In certain embodiments, the 5-6 membered nitrogen-containing heteroaryl group contains 1 or 2 nitrogen atoms.

[0036] In certain embodiments, the 5-6 membered nitrogen-containing heteroaryl group is selected from the following heteroaromatic ring-forming groups:

[0037] In certain embodiments, ring A is phenyl, which is optionally substituted with 1, 2, or 3 groups selected from the group consisting of halogen, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclyl), C1-C6 alkoxy (e.g., methoxy), hydroxy, carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, and -OCH2CH=CH2. In certain embodiments, the 3-6 membered nitrogen-containing heterocyclyl is a 5-6 membered heterocyclyl containing 1-2 nitrogen atoms and optionally an oxygen atom.

[0038] In certain embodiments, Ring A is pyridinyl, which is optionally substituted with =0.

[0039] In certain embodiments, Ring B is a naphthyl ring, optionally substituted with 1, 2, or 3 substituents.

[0040] In certain embodiments, Ring B is a benzene ring, optionally substituted with 1, 2, or 3 substituents.

[0041] In certain embodiments, the substituents of ring B are selected from the group consisting of hydrogen, halogen, nitro, nitroso, cyano, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo-C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)R b’ 、-OR c’, C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclic group, -SH, phenyl, 5-6 membered heteroaryl, -(C1-C6 alkyl)OH, -(C1-C6 alkyl)NH2, -S(C1-C3 alkyl), -S(=O)(C1-C3 alkyl), -S(=O)(=O)(C1-C3 alkyl), -S(=O)(=O)NH(C1-C3 alkyl), -NR d’ S(=O)(=O)(C1-C6 alkyl), -P(=O)R e’ R f’ , sulfonic acid, -N=NPh, -Si(C1-C3 alkyl)3; wherein, R a’ 、R b’ 、R c’ 、R d’ 、R e’ 、R f’ Each is independently selected from hydrogen, -OH, -COOH, -C(=O)H, phenyl, C1-C6 alkyl, C1-C3 alkenyl, C1-C6 alkoxy, 3-6 membered nitrogen-containing heterocyclic group, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C(=O)(C1-C6 alkyl); wherein R 3’ 、R 4’ are each independently selected from hydrogen, C 1-6 alkyl, C(═O)(C1-C6 alkyl); optionally, the 3-6 membered heterocyclic group or the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more groups selected from the group consisting of halogen, nitro, cyano, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, and C1-C6 alkoxy;

[0042] In certain embodiments, the substituents of Ring B are each independently selected from the following groups: C3-C6 saturated or unsaturated cycloalkyl, 3-6 membered heterocyclyl, phenyl, 5-6 membered heteroaryl.

[0043] In certain embodiments, the substituents of Ring B are each independently selected from the group consisting of hydrogen, halogen, nitro, cyano, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo-C1-C6 alkyl, =O, -NR 3’ R 4’ 、-CH2R a’ 、-C(=O)R b’ 、-OR c’ .

[0044] In certain embodiments, R a’ 、R b’ 、R c’ 、R d’ 、R e’ 、Rf’ 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.

[0045] In certain embodiments, R 3’ 、R 4’ are each independently selected from hydrogen, C 1-6 alkyl.

[0046] In certain embodiments, the 3-6 membered heterocyclyl is substituted by one or more groups selected from the group consisting of halogen, nitro, cyano, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, and C1-C6 alkoxy.

[0047] In certain embodiments, ring B is phenyl, which is optionally substituted with 1, 2, or 3 groups selected from the group consisting of halogen, C1-C6 alkyl, halo-C1-C6 alkyl, hydroxy, C1-C6 alkoxy, ═O, —C(═O)H, —CH2-OH, —CH2-(3-6 membered nitrogen-containing heterocyclyl), —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.

[0048] In certain embodiments, ring B is phenyl, which is optionally substituted with 1, 2, or 3 groups selected from the group consisting of halogen, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclyl), C1-C6 alkoxy (e.g., methoxy), hydroxy, carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, and -OCH2CH=CH2. In certain embodiments, the 3-6 membered nitrogen-containing heterocyclyl is a 5-6 membered heterocyclyl containing 1-2 nitrogen atoms and optionally an oxygen atom.

[0049] In certain embodiments, ring B is phenyl, which is optionally substituted with 1, 2, or 3 groups selected from C1-C6 alkyl (e.g., methyl), halogenated C1-C6 alkyl (e.g., trifluoromethyl), C1-C6 alkoxy (e.g., methoxy).

[0050] In certain embodiments, 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.

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

[0052] In certain embodiments, L is selected from the following structures: -O-CH2-, -CH2-O-, -NH-CH2-, -CH2-NH-, -CH2-CH2-.

[0053] In certain embodiments, q is 0.

[0054] 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, hydroxy, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, mercapto, -S-(C1-C6 alkyl).

[0055] In certain embodiments, T 1 Selected from hydrogen, C1-C6 alkyl.

[0056] In certain embodiments, T is selected from -CH2-.

[0057] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (I'), wherein R 1 , Q, L, Ring A, and Ring B are as defined above, respectively.

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

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

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

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

[0062] in,

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

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

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

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

[0067] 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 2 R 3 , carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, -OCH2CH=CH2; wherein, R 2 、R 3 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.

[0068] In certain embodiments, R 5 、R 6 、R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy, 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 4Each is independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is substituted by one or more C1-C6 alkyl groups.

[0069] In certain embodiments, R 5 、R 6 Each is independently selected from hydrogen, halogen, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclic group), C1-C6 alkoxy (e.g., methoxy), hydroxyl, carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, -OCH2CH=CH2. 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, C1-C6 alkyl (eg methyl), halogenated C1-C6 alkyl (eg trifluoromethyl), C1-C6 alkoxy (eg methoxy).

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

[0072] R 1 、R 2 , Q, L, R 5 、R 6 、R 7 As defined in any one of the above items respectively.

[0073] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (II"):

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

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

[0076] in,

[0077] R 1 , Q, L are as defined in any of the above items;

[0078] R 2 Selected from hydrogen or C1-C6 alkyl.

[0079] In certain embodiments, R 2 For hydrogen.

[0080] In certain embodiments, R 2 It is a methyl group.

[0081] 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.

[0082] In certain embodiments, R 5 、R 6 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy, 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 are each 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. 5 、R 6 Each is independently selected from hydrogen, halogen, -CH2-OH, -CH2-(3-6 membered nitrogen-containing heterocyclic group), C1-C6 alkoxy (such as methoxy), hydroxyl, carboxyl, -C(=O)O-(C1-C6 alkyl), -CH2-N(C1-C6 alkyl)2, -OCH2CH=CH2.

[0083] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (III'):

[0084] R 1 、R 2, Q, L, R 5 、R 6 As defined in any one of the above items respectively.

[0085] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (III"):

[0086] R 1 、R 2 , Q, L, R 5 、R 6 As defined in any one of the above items respectively.

[0087] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (IV):

[0088] in,

[0089] R 1 , Q, L are as defined in any of the above items;

[0090] R 2 Selected from hydrogen or C1-C6 alkyl.

[0091] In certain embodiments, R 2 For hydrogen.

[0092] In certain embodiments, R 2 It is a methyl group.

[0093] One of X and Y is C or CH, and the other is N or NH;

[0094] 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), -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.

[0095] In certain embodiments, R 5、R 7 Each is independently selected from: hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy, 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), -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.

[0096] In certain embodiments, R 5 is hydrogen or =O;

[0097] In certain embodiments, R 7 Selected from hydrogen, C1-C6 alkyl (eg methyl), halogenated C1-C6 alkyl (eg trifluoromethyl), C1-C6 alkoxy (eg methoxy).

[0098] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (IV'):

[0099] R 1 、R 2 , Q, L, R 5 、R 7 As defined in any one of the above items respectively.

[0100] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (IV"):

[0101] R 1 、R 2 , Q, L, R 5 、R 7 As defined in any one of the above items respectively.

[0102] In certain embodiments, the compounds of the present invention have a structure as shown in Formula (V):

[0103] Among them, R 1 , Q, L are as defined in any of the above items;

[0104] R 2 Selected from hydrogen or C1-C6 alkyl.

[0105] In certain embodiments, R 2 For hydrogen.

[0106] In certain embodiments, R 2 It is a methyl group.

[0107] 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.

[0108] In certain embodiments, R 1 For hydrogen.

[0109] In certain embodiments, Q is -O-.

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

[0111] In certain embodiments, R 5 It is hydrogen, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-N(C1-C6 alkyl)2, and -OCH2CH=CH2.

[0112] In certain embodiments, R 5 is methyl, methoxy, halogen, hydroxy, -COOH, -COOCH3, -CH2OH. In certain embodiments, R 6 It is hydrogen, -CH2-(3-6 membered nitrogen-containing heterocyclic group), -CH2-N(C1-C6 alkyl)2, and -OCH2CH=CH2.

[0113] In certain embodiments, R 6 It is methyl, methoxy, halogen, hydroxyl, -COOH, -COOCH3, -CH2OH.

[0114] In certain embodiments, R 7 is hydrogen, halogen, or C1-C6 alkoxy (e.g., methoxy).

[0115] In certain embodiments, L is -CH2-O-, -O-CH2-.

[0116] In certain embodiments, L is -NHCH2-.

[0117] The hydroxyl groups and -NH- and -NH2 in the compounds of the present invention may optionally be protected by substituents, which may be hydroxyl or amino protecting groups commonly used in the art, such as Boc (tert-butyloxycarbonyl), TBDPS (tert-butyldiphenyl), etc. The compounds thus formed are also within the scope of protection of the present invention.

[0118] The compounds of the present invention may have a structure selected from the group consisting of:

[0119] Pharmaceutical compositions, uses, and disease prevention / treatment methods

[0120] 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.

[0121] The pharmaceutical compositions of the present invention can be formulated into any pharmaceutically acceptable dosage form and administered to patients in need of such treatment by oral, parenteral, rectal, pulmonary, topical, or transdermal routes. For oral administration, conventional solid preparations such as tablets, capsules, pills, and granules can be prepared; oral liquid preparations such as oral solutions, oral suspensions, and syrups can also be prepared. Suitable fillers, binders, disintegrants, lubricants, and the like can be added to oral preparations. For parenteral administration, injections can be prepared, including injection solutions, sterile powders for injection, and concentrated solutions for injection. Injections can be produced using conventional methods in the pharmaceutical field. Additives may be omitted or added depending on the properties of the drug. For rectal administration, suppositories can be prepared. For pulmonary administration, inhalants or sprays can be prepared. For topical administration, specific dosage forms and devices can be used, or creams, gels, and the like can be prepared. For transdermal administration, patches can be prepared.

[0122] In one aspect, the present application provides the use of a compound of the present invention, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or metabolite thereof in the preparation of a medicament, which is used as a TLR7 agonist for preventing or treating a disease associated with TLR7.

[0123] In one aspect, the present application provides a method for preventing or treating a TLR7-related disease in a subject, comprising administering to a subject in need thereof an effective amount of a compound of the present invention, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystalline form, isotopically labeled form thereof, or a metabolite thereof.

[0124] In some embodiments, the disease associated with TLR7 is selected from:

[0125] (1) tumors, including but not limited to: breast cancer, melanoma, meningioma, soft tissue sarcoma, salivary gland tumor, intraspinal tumor, mediastinal tumor, brain cancer, bone cancer, penile cancer, osteosarcoma, intracranial tumor, tongue cancer, maxillary sinus cancer, thyroid cancer, malignant lymphoma, multiple myeloma, pituitary adenoma, testicular tumor, non-Hodgkin's lymphoma, bladder cancer, leukemia, gastric cancer, nasopharyngeal cancer, laryngeal cancer, oral cancer, esophageal cancer, lung cancer, kidney cancer, cervical cancer, choriocarcinoma, vulvar cancer, skin cancer, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, rectal cancer, colorectal cancer, Kaposi's sarcoma, non-melanoma skin cancer (including squamous cell carcinoma and basal cell carcinoma), hemangioma, glioma, non-Hodgkin's lymphoma (diffuse large B-cell lymphoma, B-cell non-Hodgkin's lymphoma), colon cancer, liver cancer, neuroblastoma, glioblastoma;

[0126] (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;

[0127] (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.

[0128] In the method, the compound of the present invention acts as a TLR7 agonist, and further, can act as an immunomodulator.

[0129] The compounds of the present invention have an immunoenhancing effect. Therefore, in one aspect, the present application provides the use of the compounds of the present invention, their pharmaceutically acceptable salts, prodrugs, stereoisomers, tautomers, hydrates, solvates, crystal forms, isotope labels, or metabolites thereof as vaccine adjuvants. In certain embodiments, the vaccine is used to prevent or treat diseases associated with any of the viral infections described above.

[0130] In one aspect, the present application provides use of a compound of the present invention, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance, or a metabolite thereof as a photodynamic therapy agent.

[0131] In one aspect, the present application provides the use of a compound of the present invention, a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystalline form, isotope-labeled substance, or a metabolite thereof as a conjugate drug (e.g., an antibody immunostimulatory drug conjugate, ISAC). An antibody immunostimulatory drug conjugate (ISAC) is a monoclonal antibody that combines an antibody with an immunostimulatory agent to enhance the immune response by triggering the production of proinflammatory cytokines and chemokines. For example, BDC-1001 is an ISAC that contains an antibody targeting HER2 and a TLR7 / 8 agonist and is currently in clinical phase I / II for the treatment of solid tumors.

[0132] In the present application, the subject is preferably a mammal, such as bovine, equine, porcine, canine, feline, rodent, primate, such as human.

[0133] Preparation method

[0134] 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.

[0135] Among them, -X 1 Selected from -CH2OH, -CH2OTBS, -CH2OTBDPS, -CH2OAc, -COOMe, -CHO, -X 2 is selected from -Br, -Cl, -OMs, and the remaining substituents are as described above.

[0136] Definition of terms

[0137] Unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, the laboratory procedures described herein are conventional procedures widely used in the relevant fields. To facilitate a better understanding of the present invention, definitions and explanations of relevant terms are provided below.

[0138] As used herein, the term "C1-C6 alkyl" refers to a group obtained by removing a hydrogen atom from a straight-chain or branched alkane containing 1 to 6 carbon atoms, specific examples of which include but are not limited to: methyl, ethyl, propyl, n-butyl, isobutyl, isopropyl, tert-butyl, n-pentyl, and n-hexyl. In the present invention, the preferred C1-C6 alkyl group is a C1-C4 alkyl group.

[0139] As used herein, the term "C2-C6 alkenyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon double bond and having 2-6 carbon atoms. Specific examples include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1,3-butadienyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, and 1,4-hexadienyl. In the present invention, the preferred C2-C6 alkenyl group is C2-C4 alkenyl.

[0140] As used herein, the term "C2-C6 alkynyl" refers to a straight or branched hydrocarbon group containing at least one triple bond and having 2 to 6 carbon atoms, specific examples of which include but are not limited to ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 1,3-butadiynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,4-hexadiynyl. In the present invention, the preferred C2-C6 alkynyl group is a C2-C4 alkynyl group. As used herein, the term "halogen" includes fluorine, chlorine, bromine and iodine. In the present invention, the preferred halogen group is fluorine, chlorine, bromine, and more preferably bromine.

[0141] As used herein, the term "halogenated" refers to a group or compound in which hydrogen is replaced by one or more halogen atoms, including full halogenation and partial halogenation.

[0142] As used herein, the term "alkoxy" refers to a group formed in the form of alkyl-O-.

[0143] As used herein, the term "5-6 membered heteroaryl" refers to a monocyclic aromatic group containing 5-6 ring atoms and at least one ring atom being a heteroatom, wherein the heteroatom is a nitrogen atom, an oxygen atom and / or a sulfur atom. Specific examples include, but are not limited to, furyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, pyrimidinyl, pyridazinyl, and pyrazinyl.

[0144] As used herein, the term "bicyclic heteroaryl" refers to a bicyclic group in which two rings (at least one of which has aromatic character) are fused together and at least one ring atom is a heteroatom, which is a nitrogen atom, an oxygen atom and / or a sulfur atom, which nitrogen and / or sulfur are optionally substituted with oxo. Specific examples include, but are not limited to, benzopyrrole, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzimidazole, benzotriazole, indazole, purine, indole, quinoline, pteridine, isoquinoline, quinazoline, quinoxaline, 1,8-naphthyridine, 1,5-naphthyridine, 9H-purine, 7H-purine, indane, indoline, 2,3-dihydrobenzofuran, 1,2,3,4-tetrahydronaphthalene, 1,2,3,4-tetrahydroquinoline, 3,4-dihydro-1H-benzopyran, 1,4-benzodioxane, benzomorpholine, and 1,2,3,4-tetrahydroquinoxaline.

[0145] As used herein, the term "cycloalkyl" refers to a saturated or partially saturated monocyclic alkyl group that is not aromatic, preferably having 3 to 6 ring atoms, ie, a 3-6 membered saturated cycloalkyl group and a 3-6 membered partially saturated cycloalkyl group. Specific examples of 3-6 membered saturated cycloalkyl groups include, but are not limited to, cyclopropanyl (cyclopropyl), cyclobutanyl (cyclobutyl), cyclopentanyl (cyclopentyl), and cyclohexanyl (cyclohexyl); specific examples of 3-6 membered partially saturated cycloalkyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexa-1,3-dienyl, cyclohex-1,4-dienyl, cycloheptenyl, cyclohept-1,3-dienyl, cyclohept-1,4-dienyl, cyclohept-1,3,5-trienyl, cyclooctenyl, cyclooct-1,3-dienyl, cyclooct-1,4-dienyl, cyclooct-1,5-dienyl, cyclooct-1,3,5-trienyl, and cyclooctatetraenyl.

[0146] As used herein, the term "heterocyclyl" refers to a radical derived from a saturated or partially saturated monocyclic or condensed ring compound containing 3 to 14 ring atoms and at least one heteroatom (e.g., 1, 2, 3, 4, or 5 heteroatoms) by removing one hydrogen atom. The term "3-6 membered nitrogen-containing heterocyclyl" refers to a heterocyclyl containing 3, 4, 5, or 6 ring atoms, 1, 2, or 3 of which are nitrogen atoms, and optionally further containing 1 or 2 oxygen atoms or sulfur atoms.

[0147] As used herein, the term "pharmaceutically acceptable salt" refers to (i) a salt formed by an acidic functional group (e.g., -COOH) present in the compounds provided herein with a suitable inorganic or organic cation (base), and includes, but is not limited to, alkali metal salts, such as sodium salts, potassium salts, lithium salts, etc.; alkaline earth metal salts, such as calcium salts, magnesium salts, etc.; other metal salts, such as aluminum salts, iron salts, zinc salts, copper salts, nickel salts, cobalt salts, etc.; inorganic base salts, such as ammonium salts; organic base salts, such as tert-octylamine salts, dibenzylamine salts, morpholine salts, glucosamine salts, phenylglycine alkyl ester salts, ethylenediamine salts, N-methylglucosamine salts, guanidine salts, diethylamine salts, triethylamine salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, chloroprocaine salts, procaine salts, diethanolamine salts, N-benzyl-phenethylamine salts, piperazine salts, tetramethylamine salts, tris(hydroxymethyl)aminomethane salts. and (ii) salts formed by basic functional groups (e.g., -NH2) present in the compounds provided by the present invention and appropriate inorganic or organic anions (acids), including but not limited to hydrohalides, such as hydrofluorides, hydrochlorides, hydrobromides, hydroiodides, etc.; inorganic acid salts, such as nitrates, perchlorates, sulfates, phosphates, etc.; lower alkanesulfonates, such as methanesulfonates, trifluoromethanesulfonates, ethanesulfonates, etc.; arylsulfonates, such as benzenesulfonates, p-toluenesulfonates, etc.; organic acid salts, such as acetates, malates, fumarates, succinates, citrates, tartrates, oxalates, maleates, etc.; amino acid salts, such as glycine, trimethylglycine, arginine, ornithine, glutamate, aspartate, etc.

[0148] As used herein, the term "pharmaceutically acceptable ester" refers to an ester formed between the -COOH group present in the compounds provided herein and a suitable alcohol, or an ester formed between the -OH group present in the compounds provided herein and a suitable acid (e.g., a carboxylic acid or an oxygen-containing inorganic acid). Suitable ester groups include, but are not limited to, formates, acetates, propionates, butyrates, acrylates, ethylsuccinates, stearic acid esters, or palmitates. Esters can undergo hydrolysis in the presence of an acid or base to produce the corresponding acid or alcohol.

[0149] As used herein, the term "pharmaceutically acceptable amide" refers to an amide formed between -COOH present in the compounds provided herein and a suitable amine, or an amide formed between -NH- or -NH2 present in the compounds provided herein and a suitable acid (e.g., carboxylic acid, such as acetic acid).

[0150] As used herein, the term "solvate" refers to a substance formed by the association of a compound of the present invention with a solvent molecule. The solvent may be an organic solvent (e.g., methanol, ethanol, propanol, acetonitrile, etc.). For example, the compound of the present invention may form an ethanolate with ethanol. The compound of the present invention may also form a hydrate with water.

[0151] As used herein, the term "crystalline form" refers to the crystal structure of a substance. During crystallization, various factors may alter the intramolecular or intermolecular bonding patterns, resulting in different arrangements of molecules or atoms within the crystal lattice space, forming different crystal structures. The compounds of the present invention may exist in a single crystal structure or in multiple crystal structures, i.e., they may exhibit "polymorphism." The compounds of the present invention may exist in different crystalline forms.

[0152] As used herein, the term "stereoisomer" includes conformational isomers and configurational isomers, wherein the configurational isomers primarily include cis-trans isomers and optical isomers. The compounds of the present invention may exist in the form of stereoisomers, and therefore encompass all possible stereoisomeric forms, and any combination or any mixture thereof. For example, a single enantiomer, a single diastereomer, or a mixture thereof. When a compound of the present invention contains an olefin double bond, unless otherwise specified, it includes cis-isomers and trans-isomers, and any combination thereof.

[0153] As used herein, the term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom in a molecule between two positions, such as tautomers of the enol form and the keto form. If tautomers exist for the compounds of the present invention, they may exist in the form of a single tautomer or a mixture thereof, preferably in the form of a more stable tautomer as the main form.

[0154] As used herein, the term "prodrug" refers to a compound that can be converted into the compound of the present invention by reactions such as oxidation, reduction, hydrolysis, etc. in the subject's body. The prodrug itself may or may not have the biological activity of the compound of formula (I) (e.g., regulating glycolipid metabolism activity, anti-inflammatory activity, antioxidant activity). For example, a compound of formula (I) comprising a hydroxyl or carboxyl group can be administered in the form of an ester, which is hydrolyzed in the body to form a hydroxyl compound or a carboxyl compound. Similarly, a compound of formula (I) comprising an amino group can be acylated, alkylated, or phosphorylated to form, for example, an eicosanoylamino group, an alanylamino group, or a pivaloyloxymethylamino group for administration. Further information on the use of prodrugs can be found in Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T Higuchi and W Stella) and Bioreversible Carriers in Drug Design, Pergamon Press, 1987 (ed. EB Roche, American Pharmaceutical Association). Some examples of prodrugs according to the present invention include: (i) if the compound of formula (I) contains a carboxylic acid functional group (-COOH), then its esters include, for example, replacing hydrogen with a (C1-C8)alkyl group; (ii) if the compound of formula (I) contains an alcohol functional group (-OH), then its ethers include, for example, replacing hydrogen with a (C1-C6)alkanoyloxymethyl group; and (iii) if the compound of formula (I) contains a primary or secondary amino functional group (-NH2 or -NHR, wherein R is not H), then its amides include, for example, replacing hydrogen with a (C1-C6)alkanoyloxymethyl group. 10 )alkanoyl replaces one or two hydrogens. In addition, certain compounds of formula (I) may themselves act as prodrugs of other compounds of formula (I).

[0155] As used herein, the term "pharmaceutically acceptable carrier or excipient" refers to a carrier and / or excipient that is pharmacologically and / or physiologically compatible with the subject and the active ingredient, which is well known in the art (see, e.g., Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and includes, but is not limited to, disintegrants, binders, surfactants, glidants, lubricants, pH adjusters, ionic strength enhancers, agents that maintain osmotic pressure, agents that delay absorption, diluents, antioxidants, colorants, flavorings, preservatives, taste-masking agents, etc. For example, non-limiting examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropyl cellulose, starch, pregelatinized starch, and sodium alginate. Non-limiting examples of binders include microcrystalline cellulose, gelatin, sugar, polyethylene glycol, natural and synthetic gums, polyvinyl pyrrolidone, pregelatinized starch, hydroxypropyl cellulose and hydroxypropyl methylcellulose. Non-limiting examples of diluents include lactose (monohydrate, spray-dried monohydrate, anhydrous, etc.), mannitol, xylitol, glucose, sucrose, sorbitol, microcrystalline cellulose, starch and calcium hydrogen phosphate dihydrate. Non-limiting examples of surfactants include sodium lauryl sulfate and polysorbate 80. Non-limiting examples of glidants include silicon dioxide and talc. Non-limiting examples of lubricants include magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate and a mixture of magnesium stearate and sodium lauryl sulfate. Non-limiting examples of pH regulators include, but are not limited to, phosphate buffers. Ionic strength enhancers include, but are not limited to, sodium chloride. Agents that maintain osmotic pressure include, but are not limited to, sugars, NaCl and the like. Agents that delay absorption include, but are not limited to, monostearate and gelatin. Preservatives include, but are not limited to, various antibacterial and antifungal agents such as thimerosal, 2-phenoxyethanol, parabens, chlorobutanol, phenol, sorbic acid, and the like.

[0156] As used herein, the term "prevention" refers to the method implemented in order to prevent or delay the occurrence of disease or illness or symptom in a subject. As used herein, the term "treatment" refers to the method implemented in order to obtain beneficial or required clinical results. For the purposes of the present invention, beneficial or required clinical results include but are not limited to, alleviating symptoms, narrowing the scope of the disease, stabilizing (that is, no longer worsening) the state of the disease, delaying or slowing the development of the disease, improving or alleviating the state of the disease and alleviating symptoms (no matter in part or in whole), no matter it is detectable or undetectable. In addition, "treatment" can also refer to, compared with the expected survival (if not receiving treatment), extending the survival.

[0157] As used herein, the term "effective amount" refers to an amount sufficient to obtain or at least partially obtain the desired effect. For example, a disease prevention (e.g., metabolic disease or the symptom associated with a metabolic disease) effective amount refers to an amount sufficient to prevent, stop, or delay the occurrence of a disease (e.g., metabolic disease or the symptom associated with a metabolic disease); a disease treatment effective amount refers to an amount sufficient to cure or at least partially stop the disease and its complications of a patient already suffering from the disease. It is within the capabilities of those skilled in the art to determine such an effective amount. For example, the effective amount for therapeutic use will depend on the severity of the disease to be treated, the overall state of the patient's own immune system, the patient's general condition such as age, weight and sex, the mode of administration of the drug, and other treatments administered simultaneously, etc. DETAILED DESCRIPTION

[0158] In the following examples, the structures of the compounds were determined by nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LCMS). -6 The units are given in ppm. NMR measurements were performed on a Bruker AVANCE III 500 MHz NMR instrument using deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), etc. as the solvents, and tetramethylsilane (TMS) as the internal standard.

[0159] Analytical liquid chromatography was used to monitor the reaction system and analyze the purity of key intermediates and target products. Analytical liquid chromatography system: Agilent 1260 Infinity II (equipped with 1260 Binary Pump, Vial Sampler, MCT column oven, DAD WR detector); chromatographic column: Waters XBridge BEH C18 3.5 μm, 4.6 x 150 mm.

[0160] Preparative liquid chromatography was used to separate and purify a small amount of important intermediates and the target product. The preparative liquid chromatography system used was an Agilent 1290 Infinity II (equipped with a 1290 Prep Bin Pump, a Prep ALS / FC injector / collector, and a 1260 VWD detector); a C18 reversed-phase silica gel column.

[0161] Liquid chromatography-mass spectrometry (LC-MS) was used for routine reaction system analysis and molecular weight determination of the target product. LC-MS system: Waters UPLC H-Class ultra-high performance liquid chromatography-mass spectrometry coupled with QDa; chromatographic column: Waters XBridge BEH C18 1.7 μm, 2.1 x 75 mm.

[0162] Liquid phase mobile phase ratios: Acidic conditions: A: 0.05% formic acid in water, B: acetonitrile; Alkaline conditions: A: 0.05% ammonia in water, B: acetonitrile; Neutral conditions: A: 5mM NH4HCO3 in water, B: acetonitrile. The above data are default mobile phase ratios and can be adjusted appropriately according to the properties of the measured samples during analysis and testing.

[0163] Rapid purification system was used for crude preparation and purification of compounds in conventional reaction systems. The instrument model was Biotage Isolera One, and the normal phase and reverse phase columns used were purchased from Santai Tech. The common specifications of normal phase silica gel columns are 12g, 25g, 40g, 80g, 120g, and 330g. The specific parameters are as follows: 40-63μm, The commonly used specifications of reversed-phase C18 columns are 40g, 80g, 120g and 330g, and the specific parameters are as follows: 20-45μm,

[0164] 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.

[0165] Chemical reagents The known starting materials of the present invention can be synthesized by using or according to literature, patent methods, etc. known in the art, or can be purchased from chemical reagent companies such as Alfa Aesar, Acros, Sigma-Aldrich, TCI, Aikon, Anaiji, Aladdin, J&K, Bid Pharmaceuticals, Leyan, McLean, Shaoyuan (Accela), Titan (Adamas), Xiens, and Inokane.

[0166] Abbreviations

[0167] Protecting Group Abbreviations:

[0168] Bn benzyl

[0169] Boc tert-butyloxycarbonyl

[0170] Cbz benzyloxycarbonyl

[0171] MOM methoxymethyl

[0172] PMB p-methoxybenzyl

[0173] TBDPS tert-Butyldiphenylsilyl

[0174] TBS tert-Butyldimethylsilyl

[0175] THP 2-Tetrahydropyranyl

[0176] Tos p-Toluenesulfonyl

[0177] Reagent Abbreviations:

[0178] ACN acetonitrile

[0179] AIBN Azobisisobutyronitrile

[0180] Boc2O Di-tert-butyl dicarbonate

[0181] BPO Benzoyl Peroxide

[0182] tBuONa sodium tert-butoxide

[0183] CAM Ammonium Molybdate Cerium Sulfate Color Developer

[0184] CDI N,N'-Carbonyldiimidazole

[0185] m-CPBA m-chloroperbenzoic acid

[0186] DABCO triethylenediamine

[0187] DCE 1,2-dichloroethane

[0188] DCM dichloromethane

[0189] DEAD Diethyl azodicarboxylate

[0190] DIAD Diisopropyl azodicarboxylate

[0191] DIBAL-H Diisobutylaluminum hydride

[0192] DBU 1,8-diazobisspiro[5.4.0]undec-7-ene

[0193] DHP 3,4-dihydro-2H-pyran

[0194] DIEA N,N-Diisopropylethylamine

[0195] DMAP 4-dimethylaminopyridine

[0196] DMF N,N-dimethylformamide

[0197] DMSO dimethyl sulfoxide

[0198] EA Ethyl acetate

[0199] HMPA Hexamethylphosphoramide

[0200] HATU N,N,N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphoric acid

[0201] urea

[0202] LAH lithium aluminum hydride

[0203] NBS N-bromosuccinimide

[0204] PE petroleum ether

[0205] PMA phosphomolybdic acid

[0206] PyBOP 1H-Benzotriazol-1-yloxytripyrrolidino hexafluorophosphate

[0207] TBAF Tetrabutylammonium fluoride

[0208] TBDPSCl tert-Butyldiphenylsilyl chloride

[0209] TBSCl tert-Butyldimethylsilyl chloride

[0210] TEA triethylamine

[0211] TFA trifluoroacetic acid

[0212] THF Tetrahydrofuran

[0213] TPSCl 2,4,6-Triisopropylbenzenesulfonyl chloride

[0214] TsCl p-Toluenesulfonyl chloride

[0215] TsOH p-Toluenesulfonic acid

[0216] Example

[0217] Example 1: Synthesis of Compound A-1 / A-21

[0218] 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 lithium aluminum tetrahydride (2.5 M, 95.8 μL). After the addition was complete, the entire 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. It was then 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(400MHz,DMSO-d6)δppm 8.43(s,1H),8.39-8.47(m,1H),7.86-7.94(m,2H),7.53(br s,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-1 4 -amine (2.83 mg, 7%). 1 H NMR(400MHz,DMSO-d6)δppm 7.98(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-MS m / z=360.1[M+H] + ;Calcu.=359.1.

[0219] Example 2: Synthesis of Compound A-2

[0220] 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 mixture is stirred at 25°C for 16 hours. Water (100 mL) is added to the reaction solution and extracted three times with ethyl acetate (100 mL*3). The organic phase is dried over magnesium sulfate, filtered, and concentrated. The product is purified by flash silica gel chromatography ( 330g Silica gel chromatography with 0-5% ethyl acetate / petroleum ether as eluent gave 3-((tert-butyl(dimethyl)silyl)oxymethyl)aniline (25.9 g, crude product) as a yellow oil. ESI-MS m / z = 238.0 [M+H] + ;Calcu.=237.2.

[0221] Step B: To a solution of 3-((tert-butyl(dimethyl)silyl)oxymethyl)aniline (12.0 g, 50.5 mmol) in ethanol (150 mL) was added di-tert-butyl dicarbonate (12.1 g, 55.6 mmol) at room temperature. After complete addition, the mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into water (150 mL) and extracted with ethyl acetate (200 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to yield tert-butyl N-(3-((tert-butyl(dimethyl)silyl)oxymethyl)phenyl)carbamate (17.0 g, crude) as a white solid. 1 H NMR: (400MHz, DMSO-d6) 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).

[0222] Step C: To tert-butyl N-(3-((tert-butyl(dimethyl)silyl)oxymethyl)phenyl)carbamate (6.00 g, crude) dissolved in N,N-dimethylformamide (100 mL) was added sodium hydride (1.42 g, 35.6 mmol, 60% purity) at 0°C and 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 mixture was poured into water (15 mL) and extracted with ethyl acetate (20 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford tert-butyl N-(3-((tert-butyl(dimethyl)silyl)oxymethyl)phenyl)-N-((3-(chloromethyl)phenyl)methyl)carbamate (7.70 g, crude) as a yellow oil. 1 H NMR: (400MHz, DMSO-d6) δppm 7.31-7.19(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).

[0223] Step D: To tert-butyl N-(3-((tert-butyl(dimethyl)silyl)oxymethyl)phenyl)-N-((3-(chloromethyl)phenyl)methyl)carbamate (7.60 g, 16.0 mmol) dissolved in N,N-dimethylformamide (140 mL) 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) and extracted with ethyl acetate (50 mL*3). 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). ESI-MS m / z=609.3[M+H] + ;Calcu.=608.3.

[0224] 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. The mixture was stirred at 25°C for 2 hours. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (200 ml*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated. The product was purified by flash silica chromatography ( 80g Silica gel chromatography with 0-60% ethyl acetate / petroleum ether as the eluent gave tert-butyl N-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)methyl)-N-(3-(hydroxymethyl)phenyl)carbamate (1.50 g, 37% yield over five steps) as a yellow solid. ESI-MS m / z = 495.2 [M+H] + ;Calcu.=494.2.

[0225] Step F: 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) at 0°C was added sodium hydride (24.2 mg, 606 μmol, 60% content). The mixture was stirred at 110°C for 2 hours. The mixture was filtered and then purified by preparative HPLC (column: Xtimate C18 150*40mm*10um; mobile phase: [water (FA)-MeOH]; gradient: 22%-62% B over 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%). 1H NMR: (400MHz, DMSO-d6) 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.

[0226] Example 3: Synthesis of Compound A-3

[0227] Step A: To a solution of (3-bromo-5-fluoro-phenyl)methanol (50g, 243.9mmol) in methanol (500mL), add a mixture of 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (17.84g, 24.39mmol) and triethylamine (49.35g, 487.8mmol, 67.9mL), degas with nitrogen three times, and then stir at 100°C under a carbon monoxide atmosphere for 16 hours. 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 flash silica gel chromatography ( 660g kuaisu separated Silica gel chromatography with 0-25% ethyl acetate / petroleum ether as eluent gave methyl 3-fluoro-5-(hydroxymethyl)benzoate (30 g, 67%) as a black solid. ESI-MS m / z = 185.2 [M+H] + ,Calcu.=184.0。

[0228] Step B: To a solution of methyl 3-fluoro-5-(hydroxymethyl)benzoate (16 g, 86.88 mmol) in dichloromethane (160 mL) was added triethylamine (26.37 g, 260.6 mmol, 36.28 mL). Methanesulfonyl chloride (19.34 g, 168.8 mmol, 13.1 mL) was then added at 0°C. The mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (120 mL) and extracted with dichloromethane (150 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. Methyl 3-fluoro-5-(methylsulfonyloxymethyl)benzoate (15.36 g, crude) was obtained as a white solid.

[0229] Step C: To a solution of methyl 3-fluoro-5-(methylsulfonyloxymethyl)benzoate (10 g, crude) in acetonitrile (100 mL) were added potassium carbonate (10.54 g, 76.26 mmol) and 3-(hydroxymethyl)phenol (4.78 g, 38.51 mmol) and stirred at 50°C for 16 hours. The mixture was filtered and concentrated. The product was purified by flash silica gel chromatography ( 40g quick separation The residue was purified by silica gel chromatography (eluent: 0-27% ethyl acetate / petroleum ether) to give methyl 3-fluoro-5-((3-(hydroxymethyl)phenoxy)methyl)benzoate (7.37 g, 45% yield over two steps) as a pale yellow oil. 1 H NMR(400MHz,DMSO-d6)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(br s, 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.

[0230] Step D: To a solution of methyl 3-fluoro-5-((3-(hydroxymethyl)phenoxy)methyl)benzoate (7.0 g, 24.11 mmol) in dichloromethane (70 mL) was added triethylamine (7.32 g, 72.34 mmol, 10.1 mL). Methanesulfonyl chloride (5.49 g, 47.93 mmol, 3.7 mL) was then added at 0°C. The mixture was stirred at 25°C for 4 hours. The reaction mixture was poured into water (120 mL) and extracted with dichloromethane (150 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. Methyl 3-fluoro-5-((3-(methylsulfonyloxymethyl)phenoxy)methyl)benzoate (7.1 g, crude) was obtained as a colorless oil.

[0231] 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) were added 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), followed by stirring at 50°C for 16 hours. The reaction mixture was filtered and concentrated, and the mixture was purified by flash silica gel chromatography ( 20g rapid separation Silica gel chromatography with 0-35% tetrahydrofuran / petroleum ether as the eluent gave methyl 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-5-fluoro-benzoate (1.40 g, 31% yield over two steps) as a pale yellow solid. ESI-MS m / z = 442.1 [M+H] + ;Calcu.=441.1.

[0232] Step F: To a solution of methyl 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-5-fluoro-benzoate (1.13 g, 2.56 mmol) in tetrahydrofuran (12 mL) was added diisobutylaluminum hydride (1 M, 12.8 mL) at 0°C and stirred at 0°C for 2 hours. The mixture was quenched with water (10 ml) at 0°C and extracted with ethyl acetate (12 ml*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated to give the 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.

[0233] 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, a white solid product 6. 5 -Fluorine-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%). 1HNMR(400MHz,DMSO-d6)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.

[0234] Example 4: Synthesis of Compound A-4

[0235] Step A: To a solution of (5-fluoro-1,3-phenylene)dimethanol (600 mg, 3.84 mmol) and TBDPSCl (1.16 g, 4.23 mmol) in tetrahydrofuran (20 mL) at 0°C, add imidazole (288 mg, 4.23 mmol). After complete addition, the mixture was stirred at 0°C for 3 hours. The reaction mixture was warmed to room temperature, and then water (50 mL) was added. The mixture was extracted three times with ethyl acetate (30 mL x 3). The combined organic phases were 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 was separated on a silica gel column (SiO2, PE:EA = 50:50). The reaction afforded the product (3-((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorophenyl)methanol (800 mg, 77%) as a colorless oil. ESI-MS m / z = 395.3 [M+H] + ;Calcu.=394.2.

[0236] Step B: At 0°C under N2 protection, diethyl azodicarboxylate (522 mg, 3 mmol) was added to a solution of 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) in tetrahydrofuran (20 mL). After complete addition, the mixture was stirred at 0°C overnight. The reaction solution was concentrated under reduced pressure to obtain a solid residue. The residue was separated on a silica gel column (SiO2, PE:EA = 50:50). The reaction afforded the product, methyl 5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxynicotinate (298 mg, 73%), as a pale yellow oil. ESI-MS m / z = 530.2 [M+H] + ;Calcu.=529.2。

[0237] Step C: To a solution of methyl 5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxynicotinate (265 mg, 0.5 mmol) in tetrahydrofuran (20 mL) was added lithium aluminum hydride (19 mg, 0.5 mmol) at 0°C. After the addition, the mixture was stirred at 0°C for 1 hour. After the reaction was complete, methanol (10 mL) was added under ice-cooling until no bubbles were generated. The mixture was then concentrated under reduced pressure to obtain a solid residue. The residue was separated by preparative TLC (SiO2, DCM:MeOH = 95:5). The reaction afforded the product (5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxypyridin-3-yl)methanol (150 mg, 59%) as a colorless oil. ESI-MS m / z = 502.2 [M+H] + ;Calcu.=501.2.

[0238] Step D: To a solution of (5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxypyridin-3-yl)methanol (70 mg, 0.14 mmol) and triphenylphosphine (59 mg, 0.22 mmol) in tetrahydrofuran (15 mL) at 0°C was added N-bromosuccinimide (40 mg, 0.22 mmol) in portions. After the addition, the mixture was stirred at 0°C for 2 hours. The reaction solution was concentrated under reduced pressure to give a yellow crude product, 3-(bromomethyl)-5-((3-(((tert-butyldiphenylsilyl)oxy)methyl)-5-fluorobenzyl)oxy)pyridine, which was used in the next step without purification.

[0239] Step E: To a solution of 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) in acetonitrile (10 mL) was added cesium carbonate (34.6 mg, 0.106 mmol). The mixture was stirred at room temperature for 3 hours. After the reaction was complete, the reaction solution was concentrated to obtain a solid residue. The residue was subjected to HPLC preparative 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 yielded 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, 28% yield over two steps) as a white solid. ESI-MS m / z = 653.1 [M+H] + ;Calcu.=652.2.

[0240] Step F: To a solution of 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) in acetonitrile (5 mL) at room temperature, cesium carbonate (65 mg, 0.2 mmol) and lithium iodide (27 mg, 0.2 mmol) were added. After the addition, the mixture was heated to 110°C and stirred for 18 hours. After the reaction was complete, the reaction mixture was concentrated to obtain a solid residue. The residue was subjected to HPLC preparative separation under the following 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, 5% B; 3.5-15 min, 10%-100% B; detector, UV 254 nm. This reaction yielded compound A-4, a white solid product 6. 5 -Fluorine-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, CD3OD): δppm 8.19(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.

[0241] Example 5: Synthesis of Compound A-5 / M-1

[0242] Step A: To a solution of (4-bromo-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (9 g, 20.57 mmol) in dichloromethane (150 mL) was added triethylamine (4.16 g, 41.15 mmol, 5.73 mL) and acetyl chloride (1.94 g, 24.69 mmol, 1.76 mL). After complete addition, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into ice water (150 mL) and extracted with ethyl acetate (150 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. This afforded methyl (4-bromo-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)acetate (9.8 g, crude) as a yellow oil.

[0243] Step B: To methyl (4-bromo-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)acetate (9.8 g, 20.44 mmol) dissolved in tetrahydrofuran (150 mL) was added tetrabutylammonium fluoride (1 M, 30.7 mL). After complete addition, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into ice water (150 mL) and extracted with ethyl acetate (150 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The residue was separated on a silica gel column (silica, 0-33%, petroleum ether:ethyl acetate, @ 60 mL / min) to afford methyl (4-bromo-3-((3-(hydroxymethyl)phenoxy)methyl)phenyl)acetate) as a yellow oil (3.2 g, 43% yield over two steps). 1HNMR(400MHz,DMSO-d6)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).

[0244] Step C: Methanesulfonyl chloride (2.18 g, 19.03 mmol, 1.5 mL) and triethylamine (2.66 g, 26.29 mmol, 3.7 mL) were added to methyl (4-bromo-3-((3-(hydroxymethyl)phenoxy)methyl)phenyl)acetate (3.2 g, 8.76 mmol) dissolved in dichloromethane (50 mL) at 0°C. After complete addition, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into ice water (50 mL) and extracted with ethyl acetate (50 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. This afforded methyl (4-bromo-3-((3-(methylsulfonyloxymethyl)phenoxy)methyl)phenyl)acetate (3.8 g, crude) as a yellow oil.

[0245] Step D: To methyl (4-bromo-3-((3-(methylsulfonyloxymethyl)phenoxy)methyl)phenyl)acetate (3.79 g, 8.55 mmol) dissolved in N,N-dimethylformamide (50 mL) was added 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). After the addition was complete, the mixture was stirred at 40°C for 16 hours. The reaction mixture was poured into water (50 mL) and extracted with ethyl acetate (50 mL x 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 give (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-bromo-phenyl)acetate (1.7 g, 38% yield over two steps) as a yellow solid. ESI-MS m / z = 518.0 [M+H] + ;Calcu.=517.0.

[0246] Step E: To a solution of methyl (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-bromo-phenyl)acetate (1.7 g, 3.29 mmol) in tetrahydrofuran (10 mL) at 0°C, sodium hydroxide (263 mg, 6.58 mmol) and water (10 mL) were added. The mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The residue was separated on a silica gel column (silica, 0-49% petroleum ether:ethyl acetate, @80 mL / min) to give (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-bromophenyl)methanol (1.55 g, 99%) as a white solid. ESI-MS m / z = 476.0 [M+H] + ;Calcu.=475.0.

[0247] 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 was complete, the entire mixture was stirred at 110°C for 2 hours. The reaction solution was poured into water (15 mL) and 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 -Br-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).

[0248] Step G: At 0°C, dissolve 6 6 -Br-1 1 H-4,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4To a solution of 1,4-dioxane (0.6 mL) containing 1,4-dioxane (50 mg, 114.08 μmol) of 1,4-dioxane was added sodium carbonate (36.3 mg, 342.2 μmol), water (0.2 mL), methylboronic acid (34.2 mg, 0.57 mmol), and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (8.4 mg, 11.41 μmol). After the addition was complete, the mixture was stirred at 80°C for 16 hours. The reaction solution was poured into water (1 mL) and extracted with ethyl acetate (1 mL*3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The product was then purified by reverse-phase high-performance liquid chromatography (column: C18 150*40 mm*10 μm, mobile phase: [water (formic acid)-methanol], gradient: 36%-76% B over 36 min) to obtain compound A-5, a 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(400MHz,DMSO-d6)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.

[0249] Example 6: Synthesis of Compound A-6

[0250] Step A: To methyl 3-formyl-4-hydroxybenzoate in N,N-dimethylformamide (250 mL) was added potassium carbonate (28.77 g, 208.2 mmol) and iodomethane (23.64 g, 166.5 mmol, 10.4 mL) and stirred at 25°C for 12 hours. The reaction mixture was poured into water (500 mL) and ethyl acetate (500 mL x 2) and the layers were separated. The organic layer was washed with brine (500 mL x 2) and the aqueous layer was extracted with diethyl ether. The combined organic layers were dried over magnesium sulfate, filtered, and concentrated in vacuo to afford methyl 3-formyl-4-methoxybenzoate (26 g, 96%) as a yellow solid. 1H NMR (400MHz, 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).

[0251] Step B: To a solution of methyl 3-formyl-4-methoxybenzoate (26 g, 133.89 mmol) in methanol (250 mL) at 0°C, sodium borohydride (10.21 g, 269.87 mmol) was added portionwise. The mixture was stirred at 25°C for 2 hours. The mixture was concentrated to remove the methanol, diluted with water (120 mL) at 0°C, extracted with ethyl acetate (120 mL x 3), and dried over magnesium sulfate. Filtration and concentration under reduced pressure afforded methyl 3-(hydroxymethyl)-4-methoxybenzoate (26 g, 99%) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0252] Step C: To methyl 3-(hydroxymethyl)-4-methoxybenzoate (15 g, 76.45 mmol) was added dichloromethane (150 mL), 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 x 3). The mixture was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to afford methyl 4-methoxy-3-(methylsulfonyloxymethyl)benzoate (16.67 g, 79%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) 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).

[0253] Step D: 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) were added to acetonitrile (160 mL) and stirred at 50 ° 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, the aqueous layers were combined, and extracted with diethyl ether. The combined organic layers were dried over magnesium sulfate, filtered, and concentrated in vacuo. The product was purified by flash silica gel chromatography ( 80g quick separation Silica gel column, eluting with a gradient of 0-15% ethyl acetate / petroleum ether @ 100 mL / min) to afford 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 (400MHz, DMSO-d6) 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).

[0254] 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 give a residue. Flash silica gel chromatography ( 80g quick separation Silica gel column, eluting with 0-20% tetrahydrofuran / petroleum ether gradient @100 mL / min) to give 3-((3-(hydroxymethyl)phenoxy)methyl)-4-methoxy-benzoic acid methyl ester (16 g, crude product) as a yellow oil. 1H NMR (400MHz, DMSO-d6) 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).

[0255] Step F: To a solution of methyl 3-((3-(hydroxymethyl)phenoxy)methyl)-4-methoxy-benzoate (16 g, crude) in dichloromethane (160 mL) was added methanesulfonyl chloride (18.45 g, 161.1 mmol, 12.5 mL) and triethylamine (13.39 g, 132.3 mmol, 18.4 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). The mixture was dried over magnesium sulfate, filtered and concentrated under reduced pressure to give compound methyl 4-methoxy-3-((3-(methylsulfonyloxymethyl)phenoxy)methyl)benzoate (22.8 g, crude) as a white solid. 1 H NMR(400MHz, DMSO-d6)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).

[0256] Step G: 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) were added to N,N-dimethylformamide (20 mL). The mixture was stirred at 25°C for 12 hours. The mixture was filtered and the mother liquor was concentrated and flash silica gel chromatography ( 40g quick separation Silica gel column, eluting with a 0-40% tetrahydrofuran / petroleum ether gradient at 100 mL / min, afforded the compound methyl 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-methoxybenzoate (450 mg, 22% yield over three steps) as a white solid. ESI-MS m / z = 454.1 [M+H] + ;Calcu.=453.1.

[0257] Step H: Methyl 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-methoxy-benzoate (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 diluted with H2O (50 mL) at 0°C, extracted with ethyl acetate (50 mL*3), and dried over magnesium sulfate. Filtration and concentration under reduced pressure afforded the compound (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenoxy)methyl)-4-methoxy-phenyl)methanol (450 mg, crude) as a white solid. ESI-MS m / z = 408.2 [M-OH] + ;Calcu.=425.1.

[0258] Step I: 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) was added sodium hydride (28.2 mg, 704.4 μmol, 60% purity) at 0°C. The mixture was stirred 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 layers were 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) gave 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 over two steps) as a white solid. 1H NMR (400MHz, DMSO-d6) 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=8.5 Hz, 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.

[0259] Example 7: Synthesis of Compound A-7

[0260] Step A: To a solution of 4-bromo-3-(hydroxymethyl)benzoic acid (4.8 g, 20.78 mmol) in methanol (50 mL) was added concentrated sulfuric acid (530 mg, 5.40 mmol, 288 μL) at room temperature. After complete addition, the 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 x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. Methyl 4-bromo-3-(hydroxymethyl)benzoate (4.98 g, 98%) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) 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)

[0261] Step B: At 0°C, thionyl chloride (23.79 g, 199.94 mmol, 14.52 mL) was added to a solution of methyl 4-bromo-3-(hydroxymethyl)benzoate (4.9 g, 19.99 mmol) in carbon tetrachloride (50 mL). After complete addition, the mixture was stirred at 80°C for 16 hours. The reaction mixture was poured into ice water (50 mL) and extracted with ethyl acetate (50 mL x 3). The organic phase was 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 afford methyl 4-bromo-3-(chloromethyl)benzoate (17 g, 66%) as a white solid. 1 H NMR(400MHz,DMSO-d6)δppm 8.13(s,1H),7.78-7.76(m,2H),4.84(s,2H),3.81(s,3H)

[0262] Step C: To methyl 4-bromo-3-(chloromethyl)benzoate (3.5 g, 13.28 mmol) dissolved in acetonitrile (40 mL) 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. After complete addition, the mixture was stirred at 50°C for 16 hours. The reaction mixture was evaporated, and saturated sodium bicarbonate solution (250 mL) was slowly added. The mixture was extracted with ethyl acetate (150 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The residue was separated on a silica gel column (silica, 0-4%, petroleum ether:ethyl acetate, @100 mL / min) to afford 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester (3.2 g, 52%) as a yellow oil. 1 H NMR(400MHz,DMSO-d6)δppm 8.05(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).

[0263] Step D: To a solution of 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester (3 g, 6.45 mmol) in tetrahydrofuran (30 mL) at 0°C was added diisobutylaluminum hydride (1 M, 32.23 mL). After complete addition, the mixture was stirred at 0°C for 2 hours. The reaction mixture was poured into ice water (30 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The product (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)phenyl)methanol (2.78 g, 99%) was obtained as a white solid. 1 H NMR(400MHz,DMSO-d6)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).

[0264] Step E: To a solution of (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)phenyl)methanol (1.5 g, 3.43 mmol) in dichloromethane (20 mL) at 0°C was added methanesulfonyl chloride (1.32 g, 11.52 mmol, 0.89 mL) and triethylamine (1.04 g, 10.29 mmol, 1.43 mL). After the addition was complete, the mixture was stirred at 0°C for 2 hours. The reaction mixture was poured into ice water (5 mL) and extracted with dichloromethane (5 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. This afforded the product (3-((2-bromo-5-(chloromethyl)benzyl)oxy)phenyl)methanol (600 mg, 51%) as a white solid. 1 H NMR(400MHz,DMSO-d6)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).

[0265] Step F: To (3-((2-bromo-5-(chloromethyl)benzyl)oxy)phenyl)methanol (600 mg, 1.76 mmol) dissolved in N,N-dimethylformamide (10 mL) was added 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (298 mg, 1.76 mmol) and potassium carbonate (485 mg, 3.51 mmol) at room temperature. After the addition was complete, the mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into water (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic phase was washed with saturated brine (10 mL), dried over magnesium sulfate, filtered, and concentrated. The 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 as a yellow solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0266] 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) and extracted with ethyl acetate (1 mL*3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The residue was separated on a silica gel column (silica, 0-60%, petroleum ether:ethyl acetate, @40 mL / min) to obtain a white solid product 3. 4 -Br-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

[0267] Step H: At room temperature, dissolve 3 4 -Br-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 To a solution of 1-aminobenzylamine (20 mg, 45.63 μmol) in tetrahydrofuran (0.4 mL) was added 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). After the addition was complete, the flask was evacuated and then purged with nitrogen three times. The mixture was stirred at 80°C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. Then, the product 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-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 salt (5.81 mg, 26%). 1H NMR(400MHz,DMSO-d6)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.

[0268] Example 8: Synthesis of Compound A-8 / M-2

[0269] Step A: To a solution of 2-bromo-5-(hydroxymethyl)phenol (17 g, 83.73 mmol) in tetrahydrofuran (350 mL) was added imidazole (17.10 g, 251.19 mmol) and tert-butylchlorodimethylsilane (12.62 g, 83.73 mmol, 10.30 mL). After complete addition, the mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into ice water (350 mL) and extracted with ethyl acetate (350 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The residue was separated on a silica gel column (silica, 0-5%, petroleum ether:ethyl acetate, @100 mL / min) to afford 2-bromo-5-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (19 g, 72%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0270] Step B: To 3-(bromomethyl)benzaldehyde (9.03 g, 45.38 mmol) dissolved in acetonitrile (250 mL) was added cesium carbonate (36.97 g, 113.46 mmol) and 2-bromo-5-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (12 g, 37.82 mmol). After complete addition, the mixture was stirred at 50°C for 16 hours. The reaction mixture was poured into ice water (250 mL) and extracted with ethyl acetate (250 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. 3-((2-bromo-5-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzaldehyde (15.1 g, 92%) was obtained as a white oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0271] Step C: To a solution of 3-((2-bromo-5-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzaldehyde (11.6 g, 26.64 mmol) in tetrahydrofuran (150 mL) was added tetrabutylammonium fluoride (1 M, 39.96 mL). After complete addition, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into ice water (150 mL) and extracted with ethyl acetate (150 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. 3-((2-bromo-5-(hydroxymethyl)phenoxy)methyl)benzaldehyde (12 g, crude) was obtained as a yellow oil.

[0272] Step D: To a solution of 3-((2-bromo-5-(hydroxymethyl)phenoxy)methyl)benzaldehyde (2.5 g, 7.78 mmol) in dichloromethane (50 mL) at 0°C was added methanesulfonyl chloride (1.960 g, 17.11 mmol, 1.32 mL) and triethylamine (2.36 g, 23.35 mmol, 3.25 mL). After complete addition, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (50 mL) and extracted with dichloromethane (50 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The product, methyl (4-bromo-3-((3-formylphenyl)methoxy)phenyl)methanesulfonate (3.1 g, 100%), was obtained as a white solid. 1H NMR (400MHz, DMSO-d6) 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).

[0273] Step E: To methyl (4-bromo-3-((3-formylphenyl)methoxy)phenyl)methanesulfonate (3.1 g, 7.76 mmol) dissolved in N,N-dimethylformamide (50 mL) was added 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). After the addition was complete, the mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into water (50 mL) and extracted with ethyl acetate (10 mL x 3). 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(400MHz,DMSO-d6)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).

[0274] Step F: To a solution of 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) in methanol (100 mL) was added sodium borohydride (120 mg, 3.17 mmol) and tetrahydrofuran (400 mL) at room temperature. After complete addition, the mixture was stirred at 0°C for 2 hours. The reaction mixture was poured into water (400 mL) and extracted with ethyl acetate (400 mL x 3). The organic phase was washed with brine, dried over magnesium sulfate, filtered, and concentrated. The 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, 100% yield) was obtained. ESI-MS m / z=475.9[M+H] + ;Calcu.=475.0.

[0275] Step G: Sodium hydride (328.6 mg, 8.22 mmol, 60% purity) 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 was complete, the entire mixture was stirred at 110°C for 2 hours. The reaction solution was poured into water (10 mL) and extracted with ethyl acetate (10 mL*3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The residue was separated on a silica gel column (silica, 0-49% petroleum ether:ethyl acetate, @80 mL / min) to obtain M-2, a white solid product 3. 4 -Br-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 H NMR (400MHz, DMSO-d6) 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).

[0276] Step H: Dissolve 3 4 -Br-1 1 H-4,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4To a solution of 1-aminobenzene (50 mg, 114.1 μmol) in tetrahydrofuran (0.5 mL) were added potassium, trifluoro(pyrrolidin-1-ylmethyl)borate (21.8 mg, 114.1 μmol), water (0.5 mL), sodium carbonate (36.3 mg, 342.25 μmol), (1E,4E)-1,5-diphenylpenta-1,4-dien-3-one, palladium (10.45 mg, 11.41 μmol), and 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (5.44 mg, 11.41 μmol). After the addition was complete, the mixture was stirred at 80°C for 16 h. The reaction mixture was poured into water (1 mL) and extracted with ethyl acetate (1 mL x 3). 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 (400MHz, DMSO-d6) 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(br s,2H),1.66(br s,4H),1.24(br s,2H). ESI-MS m / z=443.2[M+H] + ;Calcu.=442.2.

[0277] Example 9: Synthesis of Compound A-9 / M-3

[0278] Step A: Sodium borohydride (13.89 g, 367.2 mmol) was added to 3-hydroxy-4-methylbenzaldehyde (25 g, 183.6 mmol) dissolved in methanol (250 mL) at room temperature. The mixture was stirred at 20°C for 16 hours. The reaction solution was poured into water (250 mL) at 0°C to quench, and extracted with ethyl acetate (250 mL*3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The residue was subjected to flash silica gel chromatography ( 120g Flash lamp chromatography on a silica column with a gradient of 0-60% ethyl acetate / petroleum ether at 100 mL / min as eluent gave 5-(hydroxymethyl)-2-methylphenol (9.7 g, 38%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) δ9.14 (s, 1H), 6.96 (d, J = 7.6Hz, 1H), 6.75 (s, 1H), 6.6 1(d,J=7.5Hz,1H),5.01(t,J=5.8Hz,1H),4.36(d,J=5.8Hz,2H),2.07(s,3H).

[0279] Step B: To a solution of 5-(hydroxymethyl)-2-methylphenol (9.7 g, 70.21 mmol) in tetrahydrofuran (100 mL) at room temperature, add tert-butyldimethylsilyl chloride (10.58 g, 70.21 mmol, 8.64 mL) and imidazole (9.56 g, 140.4 mmol). After complete addition, stir the mixture at 25°C for 2 hours. Pour the reaction mixture into water (100 mL) and extract with ethyl acetate (120 mL x 3). The organic phase is dried over magnesium sulfate, filtered, and concentrated. The residue is separated on a silica gel column (SiO2, 0-9% petroleum ether:ethyl acetate, @100 mL / min) to afford 5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methylphenol (9 g, 51%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0280] Step C: Add methyl 4-bromo-3-(methylsulfonyloxymethyl)benzoate (8g, 24.76mmol) and cesium carbonate (24.20g, 74.27mmol) to a solution of 5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methylphenol (6.56g, 25.99mmol) dissolved in acetonitrile (80mL) and stir at 50°C for 16 hours. Pour the reaction mixture into water (150mL) and extract with ethyl acetate (150mL*3). Dry the organic phase with magnesium sulfate, filter and concentrate. 120g Flash chromatography column, eluting with 0-9% petroleum ether:ethyl acetate, @100 mL / min, afforded the product methyl 4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)benzoate (9.5 g, 80%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0281] Step D: To a solution of methyl 4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)benzoate (9.42 g, 19.65 mmol) dissolved in tetrahydrofuran (200 mL) was added diisobutylaluminum hydride (1 M, 98.2 mL), and the mixture was stirred at 0°C for 2 hours. The mixture was quenched with ice water (120 mL), adjusted to pH 2 with dilute hydrochloric acid (1 M), and extracted with ethyl acetate (40 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated to give the product (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)phenyl)methanol (9 g, crude) as a yellow oil.

[0282] 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). Methanesulfonyl chloride (5.68 g, 49.58 mmol, 3.8 mL) was added at 0°C, and the mixture was stirred at 25°C for 2 hours. The mixture was poured into water (120 mL) and extracted with dichloromethane (120 mL*3). The organic phase was dried over anhydrous magnesium sulfate, filtered, and concentrated to give (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methyl-phenoxy)methyl)phenyl)methyl methanesulfonate (10.6 g, crude) as a yellow oil.

[0283] 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) were added 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), and the mixture was stirred at 25° C. for 4 hours. The mixture was filtered and concentrated to give 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) as a yellow oil. ESI-MS m / z=604.1[M+H] + ;Calcu.=603.1.

[0284] Step G: To a solution of 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) 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 the product was purified by flash silica gel chromatography ( 120g Flash chromatography column, eluting with a gradient of 0-50% tetrahydrofuran / petroleum ether @ 100 mL / min) afforded the product (3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromo-phenyl)methoxy)-4-methyl-phenyl)methanol (2 g, 4.09 mmol, 21% yield over four steps) as a yellow solid. H NMR(400MHz,DMSO-d6)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).

[0285] 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 (1 g, 2.05 mmol) in N-methylpyrrolidone (10 mL) was added with sodium hydride (246 mg, 6.14 mmol, 60% content), and the mixture was stirred at 110 ° C for 8 hours. Then diluted with water (10 mL), extracted with ethyl acetate (10 mL * 3), the organic layer was dried over magnesium sulfate, filtered and concentrated, and flash silica gel chromatography ( 20g Flash chromatography column, eluent 0-50% tetrahydrofuran / petroleum ether gradient @ 80 mL / min) to obtain M-3, a 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 (400MHz, DMSO-d6) δ8.14-8.18(m,1H),7.89(s,1H),7.44-7.46(m,1H),7.08(dd,J=1.9,8.2 Hz, 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).

[0286] Step I: Sodium carbonate (70.30 mg, 663.3 μmol) was added to compound 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 (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), and the organic layer was dried over magnesium sulfate, filtered, concentrated, and then purified by reverse-phase high-performance liquid chromatography (chromatographic column: C18 150 * 40 mm * 10 um, mobile phase: [water (formic acid) - methanol], gradient: 8% ~ 48% B, 36 minutes) to obtain compound A-9, a 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 salt (25 mg, 25%). 1 H NMR(400MHz,DMSO-d6)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(br s,4H). ESI-MS m / z=457.4[M+H] + ;Calcu.=456.2.

[0287] Example 10: Synthesis of Compound A-10 / M-4

[0288] Step A: To a solution of 5-(hydroxymethyl)-2-methoxyphenol (30 g, 194.6 mmol) in tetrahydrofuran (300 mL) at room temperature, tert-butyldimethylsilyl chloride (29.33 g, 194.6 mmol, 23.9 mL) and imidazole (26.50 g, 389.2 mmol) were added. After the addition was complete, the mixture was stirred at 25°C for 2 hours. The reaction mixture was filtered and concentrated, and the residue was subjected to flash silica gel chromatography ( 120g The product was purified by flash chromatography on a silica column with a gradient of 0-9% ethyl acetate / petroleum ether at 100 mL / min as eluent to give 5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxyphenol (30 g, 57%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0289] Step B: To a solution of 5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxyphenol (6.31 g, 23.52 mmol) in acetonitrile (80 mL) at room temperature, add cesium carbonate (24.20 g, 74.27 mmol) and methyl 4-bromo-3-(methylsulfonyloxymethyl)benzoate (8 g, 24.76 mmol). After complete addition, stir the mixture at 50°C for 16 hours. Pour the reaction mixture into water (150 mL) at 0°C and extract with ethyl acetate (150 mL x 3). Dry the organic phase over magnesium sulfate, filter, and concentrate. The residue was separated on a silica gel column (silica, 0-10% petroleum ether:ethyl acetate, @100 mL / min) to give 4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxy-phenoxy)methyl)benzoate (10.1 g, 82%) as a yellow oil. 1 H NMR(400MHz,DMSO-d6)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,1H),5.16(s,2H),4.60(s,2H),3.85(s,3H),3.79(s,3H),0.85(s,9H),0.02(s,6H).

[0290] Step C: 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) was added diisobutylaluminum hydride (1 M, 101 mL) dropwise at 0°C 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 x 3). The organic layer was dried over magnesium sulfate, filtered, and concentrated. The product (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxy-phenoxy)methyl)phenyl)methanol (10.35 g, crude) was obtained as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0291] Step D: 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) in dichloromethane (120 mL) was added triethylamine (6.72 g, 66.42 mmol, 9.3 mL), and at 0°C, methanesulfonyl chloride (6.37 g, 55.61 mmol, 4.3 mL) was added, and the mixture was stirred at 25°C for 2 hours. The mixture was diluted with water (120 mL) and extracted with dichloromethane (20 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated to give the yellow solid product (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxy-phenoxy)methyl)phenyl)methyl methanesulfonate (11.9 g, crude).

[0292] Step E: 6-Chloro-2H-pyrazolo[3,4-d]pyrimidin-4-amine (3.70 g, 21.81 mmol) was added to a solution of (4-bromo-3-((5-((tert-butyl(dimethyl)silyl)oxymethyl)-2-methoxy-phenoxy)methyl)phenyl)methyl methanesulfonate (11.9 g, crude) in DMF (120 mL), followed by the addition of potassium carbonate (9.04 g, 65.44 mmol), and the mixture was stirred at 25°C for 7 hours. The mixture was filtered and concentrated to give 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) as a yellow oil. ESI-MS m / z = 620.1 [M+H] + ;Calcu.=619.1。

[0293] Step F: To a solution of 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 chromatography with a 0-68% tetrahydrofuran / petroleum ether gradient @ 100 mL / min as the eluent gave the 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, 40% yield over four steps) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0294] 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 (1 g, 1.98 mmol) in N-methylpyrrolidone (10 mL), sodium hydride (238 mg, 5.94 mmol, 60% content) was added and stirred at 110°C for 16 hours. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 ml * 2). The organic layer was dried over magnesium sulfate, filtered and concentrated, and flash silica gel chromatography ( 12g Flash chromatography column, eluent: 0-51% tetrahydrofuran / petroleum ether gradient @ 60 mL / min) to obtain M-4, an 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.

[0295] 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, followed by the addition of potassium trifluoro(pyrrolidin-1-ylmethyl)borate (40.8 mg, 213.5 μmol) and 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (10.2 mg, 21.35 μmol). Then, under nitrogen protection, tris(dibenzylideneacetone)dipalladium (19.6 mg, 21.35 μmol) was added. The mixture was stirred at 80°C for 10 hours, then diluted with water (3 mL) and extracted with ethyl acetate (3 mL*3). The organic layer was dried over magnesium sulfate, filtered and concentrated, and then subjected to reverse-phase high performance liquid chromatography (chromatographic column: C18 150*40 mm*10 um, mobile phase: [water (formic acid)-methanol], gradient: 32% to 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(400MHz,DMSO-d6)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),1.66-1.70(m,4H). ESI-MS m / z=473.4[M+H] + ;Calcu.=472.2.

[0296] Example 11: Synthesis of Compound A-11 / M-5

[0297] Step A: To a solution of 3-fluoro-5-hydroxybenzaldehyde (25 g, 178.4 mmol) in methanol (250 mL) was added sodium borohydride (13.66 g, 361.1 mmol). The mixture was stirred at 0°C for 2 hours. The mixture was concentrated to remove the methanol, diluted with water (120 mL) at 0°C, extracted with ethyl acetate (120 mL x 3), and dried over magnesium sulfate. Filtration and concentration under reduced pressure afforded 3-fluoro-5-(hydroxymethyl)phenol (19.58 g, 77%) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0298] Step B: To a solution of 3-fluoro-5-(hydroxymethyl)phenol (19.58 g, 137.8 mmol) in tetrahydrofuran (200 mL) were added tert-butyldimethylsilyl chloride (20.76 g, 137.8 mmol, 17 mL) and imidazole (18.76 g, 275.5 mmol) and stirred 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 x 3). The mixture was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to afford 3-(((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenol (29.5 g, 115.06 mmol, 84% yield) as a white solid.1 H NMR (400MHz, DMSO-d6) 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).

[0299] Step C: To a solution of methyl 4-bromo-3-formylbenzoate (25 g, 102.86 mmol) in methanol (250 mL) was added sodium borohydride (8.61 g, 227.6 mmol) portionwise. The mixture was stirred at 0°C for 2 hours. The reaction mixture was concentrated to remove the methanol, diluted with water (120 mL) at 0°C, extracted with ethyl acetate (120 mL x 3), and dried over magnesium sulfate. Filtration and concentration under reduced pressure afforded methyl 4-bromo-3-(hydroxymethyl)benzoate (18 g, 71%) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0300] Step D: To dichloromethane (200 mL) was added methyl 4-bromo-3-(hydroxymethyl)benzoate (18 g, 73.45 mmol) and triethylamine (22.30 g, 220.4 mmol, 30.7 mL). Methanesulfonyl chloride (21.23 g, 185.33 mmol, 14.34 mL) was added dropwise at 0°C and stirred at 25°C for 2 hours. The reaction mixture was quenched with ice water (200 mL) at 0°C and extracted with dichloromethane (100 mL x 2). The mixture was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to afford methyl 4-bromo-3-(chloromethyl)benzoate (19 g, 98%) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ = 8.14 (s, 1H), 7.86-7.91 (m, 2H), 5.38 (s, 2H), 3.88-3.87 (m, 3H).

[0301] 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) afforded methyl 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)benzoate (9.2 g, 77%) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0302] Step F: To a solution of methyl 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)benzoate (9.2 g, 19.03 mmol) in tetrahydrofuran (100 mL) was added diisobutylaluminum hydride (1 M, 95 mL). The mixture was stirred at 0°C for 2 hours. The mixture was quenched with ice water (200 mL) and adjusted to pH 8 with 1N hydrochloric acid. The mixture was extracted with ethyl acetate (200 mL x 2). The organic layer was dried over magnesium sulfate, filtered, and concentrated. This afforded a yellow oil (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl ester)methanol (10.63 g, crude product).

[0303] 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) were added methanesulfonyl chloride (6.69 g, 58.40 mmol, 4.5 mL) and triethylamine (4.72 g, 46.68 mmol, 6.50 mL). The mixture was stirred at 0°C for 2 hours, quenched with water (100 mL), and extracted with dichloromethane (100 mL*3). The mixture was dried over magnesium sulfate, filtered, and concentrated 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.

[0304] Step H: To N,N-dimethylformamide (100 mL) were added 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) and stirred at 25°C for 12 hours. The reaction mixture 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.

[0305] Step I: 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 (15 g, crude) in tetrahydrofuran (130 mL) was added tetrabutylammonium fluoride (1 M, 37.07 mL, 1.5 eq). The reaction mixture was stirred at 25°C for 12 hours. Filtered and the mother liquor was concentrated. The residue was purified by flash silica gel chromatography ( 80g Purification by silica flash lamp chromatography with a 0-40% tetrahydrofuran / petroleum ether gradient at 100 mL / min afforded (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

[0306] Step J: To a solution of N-methylpyrrolidone (100 mL) was added (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). 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 was performed by silica flash lamp chromatography (eluent: 0-60% THF / petroleum ether gradient @ 100 ml / min) to afford M-5 as a white solid 3 4 -Br-6 5 -Fluorine-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.

[0307] Step K: Add 3 4 -Br-6 5 -Fluorine-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), tris(dibenzylideneacetone)dipalladium (20.1 mg, 21.92 μmol), and sodium carbonate (69.7 mg, 657.5 μmol) were purged with nitrogen three times and then stirred at 80°C under a nitrogen atmosphere 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-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 (4.3 mg, 4%). 1H NMR(400MHz,DMSO-d6)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),7.05-7.16(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.

[0308] Example 12: Synthesis of Compound A-12

[0309] Step I: At room temperature, dissolve 3 4 -Br-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 To a solution of 4-aminobenzene (20 mg, 45.63 μmol) in tetrahydrofuran (0.4 mL) was added water (0.2 mL), potassium (morpholin-4-yl)methyltrifluoroborate (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). After the addition was complete, the flask was evacuated and then purged with nitrogen three times. The mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The product 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-12, a white solid product 3 4 -(morpholinomethyl)-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%). 1H NMR (400MHz, DMSO-d6) 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.

[0310] Example 13: Synthesis of Compound A-13 / A-16

[0311] Step A: To methyl 4-bromo-3-formylbenzoate (10 g, 41.14 mmol) dissolved in methanol (100 mL) was added sodium borohydride (1.87 g, 49.37 mmol) at 0°C. After complete addition, the mixture was stirred at 0°C for 2 hours. The reaction mixture was poured into ice water (100 mL) and extracted with ethyl acetate (100 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. Methyl 4-bromo-3-(hydroxymethyl)benzoate (10 g, 99%) was obtained as a yellow oil. 1 H NMR (400MHz, DMSO-d6) δppm 8.10-8.14 (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).

[0312] Step B: To a solution of methyl 4-bromo-3-(hydroxymethyl)benzoate (24.00 g, 97.93 mmol) in chloroform (240 mL) at 0°C, add thionyl chloride (116.51 g, 979.3 mmol, 71.1 mL). After complete addition, stir the mixture at 80°C for 16 hours. Pour the reaction mixture into ice water (250 mL) and extract with ethyl acetate (250 mL x 3). The organic phase is dried over magnesium sulfate, filtered, and concentrated. The residue is separated on a silica gel column (silica, 0-10% petroleum ether:ethyl acetate, @100 mL / min) to afford methyl 4-bromo-3-(chloromethyl)benzoate (17 g, 66%) as a white solid. 1 H NMR (400MHz, DMSO-d6) δppm 8.20 (d, J = 1.3Hz, 1H), 7.82-7.88 (m, 2H), 4.92 (s, 2H), 3.88 (s, 3H).

[0313] Step C: To methyl 4-bromo-3-(chloromethyl)benzoate (17 g, 64.51 mmol) dissolved in acetonitrile (17 mL) 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. After complete addition, the mixture was stirred at 50°C for 16 hours. The reaction mixture was poured into ice water (250 mL) and extracted with ethyl acetate (150 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The residue was separated on a silica gel column (silica, 0-4% petroleum ether:ethyl acetate, @100 mL / min) to afford 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester (3.2 g, 52%) as a yellow oil. 1 H NMR(400MHz,DMSO-d6)δppm 8.05(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).

[0314] Step D: To a solution of 4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester (20 g, 42.97 mmol) in tetrahydrofuran (200 mL) was added diisobutylaluminum hydride (1 M, 214.85 mL) at 0°C. After complete addition, the mixture was stirred at 0°C for 2 hours. The reaction mixture was poured into ice water (300 mL) and extracted with ethyl acetate (300 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The product (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)phenyl)methanol (18 g, 96%) was obtained as a white oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0315] Step E: To a solution of (4-bromo-3-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)phenyl)methanol (10 g, 22.86 mmol) in dichloromethane (100 mL) at 0°C was added methanesulfonyl chloride (7.39 g, 64.51 mmol, 4.99 mL) and triethylamine (6.94 g, 68.58 mmol, 9.55 mL). After the addition was complete, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into ice water (100 mL) and extracted with dichloromethane (100 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. 4-bromo-3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzylsulfonate (11 g, 93%) was obtained as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0316] Step F: To a solution of 4-bromo-3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzylsulfonate (11 g, 21.34 mmol) in N,N-dimethylformamide (250 mL) at room temperature was added 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). After the addition was complete, the mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into water (500 mL) and extracted with ethyl acetate (500 mL x 3). The organic phase was rinsed with brine, 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。

[0317] Step G: To a solution of 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) in tetrahydrofuran (160 mL) at 0°C was added tetrabutylammonium fluoride (1 M, 31.6 mL). After complete addition, the mixture was stirred at 25°C for 2 h. The reaction mixture was poured into ice water (160 mL) and extracted with ethyl acetate (160 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The residue was separated on a silica gel column (silica, 0-41% petroleum ether:ethyl acetate, @100 mL / min) to give (3-((5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-bromobenzyl)oxy)phenyl)methanol (4 g, 40%) as a yellow solid. ESI-MS m / z = 475.8 [M+H] + ;Calcu.= 475.0.

[0318] Step H: At room temperature, sodium hydride (379 mg, 9.48 mmol, 60% purity) was added to 1-methylpyrrolidin-2-one (20 mL) dissolved with (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). After the addition was complete, the mixture was stirred at 110°C for 2 hours. The reaction solution was poured into ice water (160 mL) and extracted with ethyl acetate (160 mL*3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to obtain A-6, a yellow solid product 3. 4 -Br-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidin-3,6(1,3)-dibenzocyclooctan-14-amine (600 mg, 43%). ESI-MS m / z = 439.9 [M+H] + ;Calcu.=439.0.

[0319] Step I: Dissolve 3 4 -Br-1 1To a solution of H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidin-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-dicyclohexylphosphino-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 purged with nitrogen three times. The mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The product was then 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-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 (400MHz, DMSO-d6) 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=7.4 Hz,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.

[0320] Example 14: Synthesis of Compound A-14

[0321] Step A: Dissolve 3 4 -Br-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4To a solution of 1,3-dimethylaminomethyltrifluoroborate (30 mg, 68.45 μmol) in tetrahydrofuran (0.4 mL) was added water (0.2 mL), potassium (dimethylamino)methyltrifluoroborate (11.3 mg, 68.45 μmol), 2-dicyclohexylphosphino-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 refilled with nitrogen three times. The mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The product was then 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 salt (8.32 mg, 28%). 1 HNMR(400MHz,DMSO-d6)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.

[0322] Example 15: Synthesis of Compound A-15

[0323] Step A: Sodium borohydride (0.45 g, 11.9 mmol) was added portionwise to methyl 4-fluoro-3-formylbenzoate (6.0 g, 32.9 mmol) in methanol (160 mL) at 0°C. After complete addition, the mixture was stirred at 25°C for 2 hours. Water (100 mL) was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (200 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to yield methyl 4-fluoro-3-(hydroxymethyl)benzoate (5.90 g, 97%). 1H NMR: (400MHz, DMSO-d6) 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.

[0324] Step B: To methyl 4-fluoro-3-(hydroxymethyl)benzoate (5.80 g, 31.5 mmol) dissolved in dichloromethane (100 mL) at 0°C, triethylamine (9.56 g, 94.5 mmol) and methanesulfonyl chloride (5.70 g, 49.8 mmol) were added. After complete addition, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (50 mL) and extracted with dichloromethane (100 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford methyl 4-fluoro-3-(methylsulfonyloxymethyl)benzoate (5.60 g, crude) as a yellow oil.

[0325] 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 methyl 4-fluoro-3-(methylsulfonyloxymethyl)benzoate (5.60 g, crude product) in acetonitrile (100 mL), and the mixture was stirred at 50°C for 16 hours. The reaction mixture was filtered and concentrated. The product was purified by flash silica gel chromatography ( 80g The product was purified by chromatography on a silica gel column with 0-3% ethyl acetate / petroleum ether as the eluent to give methyl 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl)-4-fluorobenzoate (5.81 g, 46% yield over two steps) as a yellow oil.

[0326] Step D: To methyl 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-4-fluorobenzoate (5.80 g, 14.3 mmol) dissolved in tetrahydrofuran (100 mL) at 0°C was added diisobutylaluminum hydride (1 M, 57.4 mL). 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 3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-4-fluorophenyl)methanol (5.38 g, crude) as a yellow oil. 1H NMR: (400MHz, DMSO-d6) 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).

[0327] Step E: 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) at 0°C, triethylamine (1.61 g, 15.9 mmol) and methanesulfonyl chloride (1.24 g, 10.8 mmol) were added. After the addition was complete, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (30 mL) and extracted with dichloromethane (50 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford tert-butyl-((3-((5-(chloromethyl)-2-fluoro-phenyl)methoxy)phenyl)formyl)-dimethylsilane (2.55 g, crude) as a yellow oil.

[0328] Step F: To 6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-amine (429 mg, 2.53 mmol) dissolved in N,N-dimethylformamide (20 mL) was added 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) at room temperature, and the mixture was stirred at 25°C for 16 hours. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (20 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated to give (3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-4-fluorophenyl)methanol (1.30 g, crude) as a yellow oil. ESI-MS m / z = 528.3 [M+H] + ;Calcu.=527.2.

[0329] 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) and extracted with ethyl acetate (20 ml * 3). The organic layer was dried over magnesium sulfate, filtered, and concentrated. The product was purified by flash silica gel chromatography ( 20g Silica gel chromatography with 0-47% ethyl acetate / petroleum ether as the eluent gave the product (3-((5-((4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-fluorobenzyl)oxy)phenyl)methanol (430 mg, 43%) as a yellow solid. ESI-MS m / z = 413.9 [M+H] + ;Calcu.=413.1.

[0330] Step H: Sodium hydride (43.5 mg, 1.09 mmol, 60% content) 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) in N-methylpyrrolidone (3 mL) at 0°C, and the mixture was stirred at 110°C for 2 hours. The mixture was filtered and then purified by preparative high performance liquid chromatography (formic acid conditions, 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 -Fluorine-1 1 H-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-d6) 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.

[0331] Example 16: Synthesis of Compound A-17

[0332] Step A: To N,N-dimethylformamide (200 mL) was added methyl 3-formyl-4-hydroxybenzoate (10 g, 55.51 mmol), potassium carbonate (11.51 g, 83.26 mmol), and iodomethane (9.45 g, 66.61 mmol, 4.15 mL). The reaction was stirred at 25°C for 12 hours. The reaction was poured into water (500 mL) and ethyl acetate (500 mL x 2), and the layers were separated. The organic layer was washed with brine (500 mL x 2). The aqueous layers were combined and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, filtered, and concentrated in vacuo to afford methyl 3-formyl-4-methoxybenzoate (12 g, crude) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0333] Step B: To a solution of methyl 3-formyl-4-methoxybenzoate (12 g, crude) in methanol (120 mL) was added sodium borohydride (2.81 g, 74.16 mmol) at 0°C. The mixture was stirred at 0°C for 2 hours. The reaction mixture was diluted with water (120 mL) at 0°C, extracted with ethyl acetate (120 mL x 3), and dried over magnesium sulfate. Filtration and concentration under reduced pressure afforded methyl 3-(hydroxymethyl)-4-methoxybenzoate (7.69 g, 70% yield over two steps) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0334] Step C: Methanesulfonyl chloride (4.4 g, 38.41 mmol, 3.0 mL) was added to a solution of methyl 3-(hydroxymethyl)-4-methoxybenzoate (3 g, 15.29 mmol) and triethylamine (3.09 g, 30.58 mmol, 4.3 mL) in dichloromethane (50 mL) at 0°C. The mixture was stirred at 25°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). The mixture was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to afford methyl 3-(chloromethyl)-4-methoxybenzoate (2.7 g, 82%) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0335] Step D: To acetonitrile (80 mL) was added 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). The mixture was stirred at 50°C for 12 hours. The mixture was filtered and the mother liquor was concentrated at 50°C. The product was purified by flash silica gel chromatography ( 40 grams Flash lamp chromatography on a silica column with a gradient of 0-20% ethyl acetate / petroleum ether at 100 mL / min as eluent afforded methyl 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl)-4-methoxybenzoate (3.82 g, 89%) as a yellow oil. 1 H NMR(400MHz,DMSO-d6)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).

[0336] Step E: To a solution of methyl 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl)-4-methoxybenzoate (1.82 g, 4.37 mmol) in tetrahydrofuran (20 mL) was added diisobutylaluminum hydride (1 M, 21.84 mL). The mixture was stirred at 0°C for 2 hours. The reaction mixture was quenched with ice water (50 mL), 1N hydrochloric acid was added to pH = 8, and extraction was performed with ethyl acetate (50 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated. This afforded (3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl)-4-methoxyphenyl)methanol (1.6 g, 94%) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0337] 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. The mixture was stirred 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). The mixture was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give (3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-4-methoxyphenyl)methanesulfonate (1.0 g, 52%) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0338] Step G: To a solution of methyl (3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-4-methoxyphenyl)methanesulfonate (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) was added potassium carbonate (188.8 mg, 1.37 mmol) and stirred at 25°C for 12 hours. The reaction mixture was filtered and the mother liquor was concentrated at 60°C to give 1-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-4-methoxyphenyl)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.

[0339] 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) was added tetrabutylammonium fluoride (1 M, 799.8 μL) and stirred 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 Purification by flash chromatography using a 0-27% tetrahydrofuran / petroleum ether gradient at 100 mL / min as the eluent yielded 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.

[0340] 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) was added sodium hydride (18.60 mg, 464.93 μmol, 60% purity) at 0°C. 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: C18150*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, 1.5% yield over three steps). 1 H NMR (400MHz, CD3OD) 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),6.9 8(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.

[0341] Example 17: Synthesis of Compound A-18

[0342] Step A: To acetonitrile (100 mL) dissolved with 5-(hydroxymethyl)-2-methoxyphenol (7.03 g, 45.62 mmol) was added potassium carbonate (12.61 g, 91.24 mmol) and 3-(bromomethyl)benzaldehyde (9.08 g, 45.62 mmol) at room temperature. The mixture was stirred at 50°C for 5 hours. The reaction mixture was filtered and concentrated, and the residue was subjected to flash silica gel chromatography ( 40g Flash chromatography with a gradient of 0-35% ethyl acetate / petroleum ether at 100 mL / min as eluent gave 3-((5-(hydroxymethyl)-2-methoxy-phenoxy)methyl)benzaldehyde (6.8 g, 55%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0343] Step B: To a solution of 3-((5-(hydroxymethyl)-2-methoxy-phenoxy)methyl)benzaldehyde (6.8 g, 24.97 mmol) in dichloromethane (100 mL) at room temperature was added triethylamine (7.58 g, 74.92 mmol, 10.43 mL). Methanesulfonyl chloride (5.82 g, 50.81 mmol, 3.93 mL) was then added dropwise at 0°C. After complete addition, the mixture was stirred at 25°C for 4 hours. The reaction mixture was poured into water (120 mL) and extracted with ethyl acetate (120 mL x 3). The organic phase was dried over anhydrous magnesium sulfate, filtered, and concentrated. This afforded (3-((3-formylphenyl)methoxy)-4-methoxyphenyl)methyl methanesulfonate (5.26 g, crude) as a white solid.

[0344] Step C: 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) was added 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) and stirred at 40°C for 5 hours. The mixture was filtered and concentrated. The residue was subjected to 20g Flash chromatography (eluent: 0-27% petroleum ether:tetrahydrofuran, @100 mL / min) afforded 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, 25% yield over two steps) as a yellow solid. ESI-MS m / z = 424.1 [M+H] + ;Calcu.=423.1.

[0345] Step D: To a solution of 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) was added sodium borohydride (0.250 g, 6.61 mmol), and the mixture was stirred at 0°C for 10 hours. The mixture was quenched with ice water (10 mL) and extracted with ethyl acetate (40 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated to give the product (3-((5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-methoxy-phenoxy)methyl)phenyl)methanol (700 mg, 87%) as a yellow solid. ESI-MS m / z = 426.1 [M+H] + ;Calcu.=425.1.

[0346] 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) and extracted with ethyl acetate (1 mL*3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. Then, the product was purified by reverse-phase high-performance liquid chromatography (column: C18 150*40 mm*10 um, 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 (400MHz, DMSO-d6) 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.

[0347] Example 18: Synthesis of Compound A-19 / A-20

[0348] Step A: Dissolve 34 -Br-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 To a solution of 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (33.4 mg, 45.63 μmol) of 2-amino-2-nitropropane (200 mg, 456.33 μmol) in methanol (100 mL) and N,N-dimethylformamide (5 mL) was added 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (33.4 mg, 45.63 μmol) and triethylamine (92.4 mg, 912.7 μmol, 127 μL). After the addition was complete, the mixture was stirred at 100°C for 16 hours under a carbon monoxide (1 MPa) atmosphere. Water (200 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (200 mL*3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The product A-20 was then purified by preparative HPLC (column: Xtimate C18 150*40 mm*10 μm; mobile phase: [water (FA)-MeOH]; gradient: 32%-72% B in 36 minutes) to obtain a white solid product A-20. 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(400MHz,DMSO-d6)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.

[0349] Step B: At 0℃, 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. It was then 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 (400MHz, DMSO-d6) 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.

[0350] Example 19: Synthesis of Compound A-22 / M-6

[0351] Step A: Dissolve 3 4 -Br-1 1 H-4,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4 -amine (200 mg, 456.3 μmol) in methanol (5 mL) was added with N, N-dimethylformamide (5 mL), 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (33.4 mg, 45.63 μmol) and triethylamine (92.3 mg, 912.6 μmol, 127 μL). After the addition was complete, the entire mixture was stirred at 100 ° C for 48 hours under a carbon monoxide (1 MPa) atmosphere. The reaction solution was poured into water (10 mL) and extracted with ethyl acetate (10 mL*3). The organic phase was dried over 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.

[0352] Step B: At 0℃, 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 (1 M, 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) and extracted with ethyl acetate (1 mL * 3). The organic phase was dried over magnesium sulfate, filtered and concentrated. It was then purified by reverse phase high performance liquid chromatography (chromatographic column: chromatographic column: C18 150 * 30 mm * 5 um, mobile phase: [water (formic acid) - acetonitrile], gradient: 2% -42% B over 25 min) 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 (400MHz, DMSO-d6) 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.

[0353] Example 20: Synthesis of Compound A-23

[0354] Step A: To N,N-dimethylformamide (100 mL) was added 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) and stirred at 25°C for 12 hours. The reaction was poured into water (500 mL) and ethyl acetate (500 mL x 2) and the layers were separated. The organic layer was washed with brine (500 mL x 2), and the combined aqueous layers were extracted with ether. The organic layers were combined, dried over magnesium sulfate, filtered, and concentrated in vacuo to afford 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl ester)benzaldehyde (12 g, 80%) as a yellow oil. 1 H NMR(400MHz,DMSO-d6)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).

[0355] Step B: To a solution of 3-((3-(((tert-butyldimethylsilyl)oxy)methyl)-5-fluorophenoxy)methyl)benzaldehyde (11.5 g, 30.71 mmol) in tetrahydrofuran (150 mL) was added tetrabutylammonium fluoride (1 M, 46 mL), and the reaction was stirred at 25°C for 12 hours. The reaction was quenched with water (100 mL) at 0°C. The reaction mixture was extracted with ethyl acetate (100 mL*3). The mixture was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-((3-fluoro-5-(hydroxymethyl)phenoxy)methyl)benzaldehyde (7.2 g, 90%) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0356] Step C: To dichloromethane (100 mL) 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. The temperature was lowered to 0°C, and methanesulfonyl chloride (8.22 g, 71.76 mmol, 5.6 mL) was added. The reaction mixture was stirred at 25°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 x 3). The mixture was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to afford methyl 3-(chloromethyl)-4-methoxybenzoate (10.8 g, crude) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0357] Step D: To a solution of methyl 3-(chloromethyl)-4-methoxybenzoate (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 mixture was filtered and the mother liquor was concentrated. The residue was purified by flash silica gel chromatography ( 40 grams Silica flash lamp chromatography with a 0-40% tetrahydrofuran / petroleum ether gradient at 100 mL / min afforded 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.

[0358] 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) was added sodium borohydride (55.1 mg, 1.46 mmol) portionwise at 0°C. The mixture was stirred at 0°C for 2 hours. The mixture was diluted with water (50 mL) at 0°C, extracted with ethyl acetate (50 mL*3), and dried over magnesium sulfate. Filtration and concentration under reduced pressure gave (3-((3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-fluorophenoxy)methylphenyl)methanol (400 mg, crude) as a white solid. ESI-MS m / z = 414.1 [M+H] + ;Calcu.=413.1.

[0359] Step F: 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) was added sodium hydride (29.0 mg, 724.9 μmol, 60% purity) 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 -Fluorine-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 over two steps). 1 H NMR(400MHz,DMSO-d6)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.

[0360] Example 21: Synthesis of Compound A-24

[0361] Step A: A mixture of 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (8.51 g, 11.63 mmol) and triethylamine (23.53 g, 232.51 mmol, 32.36 mL) was added to a solution of 3-bromo-5-methoxy-benzaldehyde (25 g, 116.26 mmol) in methanol (300 mL), degassed with nitrogen three times, and then stirred 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 flash silica gel chromatography ( 80g The residue was chromatographed on a silica gel column with 0-8% ethyl acetate / petroleum ether as the eluent to obtain methyl 3-formyl-5-methoxybenzoate (21.6 g, 96%) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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。

[0362] Step B: To a solution of methyl 3-formyl-5-methoxybenzoate (21.5 g, 110.72 mmol) in methanol (210 mL) was added sodium borohydride (30.09 g, 795.35 mmol) and stirred at 0°C for 4 hours. The reaction mixture was poured into water (120 mL) and extracted with ethyl acetate (120 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The product, methyl 3-(hydroxymethyl)-5-methoxybenzoate (20 g, 101.94 mmol, 92% yield), was obtained as a white oil. 1 H NMR (400MHz, DMSO-d6) 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.

[0363] Step C: To a solution of 3-(hydroxymethyl)-5-methoxy-benzoic acid methyl ester (10 g, 50.97 mmol) in carbon tetrachloride (100 mL) was added thionyl chloride (60.64 g, 509.7 mmol, 37 mL) and stirred at 80°C for 16 hours. The mixture was filtered and concentrated. The product was purified by flash silica gel chromatography ( 40g The residue was chromatographed on a silica gel column with 0-3% ethyl acetate / petroleum ether as eluent to obtain 3-(chloromethyl)-5-methoxybenzoic acid methyl ester (5.3 g, 48%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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.

[0364] Step D: To a solution of 3-(chloromethyl)-5-methoxy-benzoic acid methyl ester (5.2 g, 24.23 mmol) in acetonitrile (50 mL) were added cesium carbonate (23.68 g, 72.68 mmol) and compound 3-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (5.78 g, 24.23 mmol), and the mixture was stirred at 50°C for 16 hours. The mixture was filtered and concentrated. The product was purified by flash silica gel chromatography ( 120 grams Chromatography on a silica column with a gradient of 0-5% ethyl acetate / petroleum ether at 100 mL / min as the eluent afforded the product 3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-5-methoxy-benzoic acid methyl ester (7.5 g, 74%) as a colorless oil. 1 H NMR(400MHz,DMSO-d6)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).

[0365] Step E: To a solution of methyl 3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-5-methoxy-benzoate (4 g, 9.60 mmol) in tetrahydrofuran (80 mL) was added diisobutylaluminum hydride (1 M, 48 mL) at 0°C, followed by stirring at 0°C for 2 hours. The mixture was quenched with ice water (80 mL) and extracted with ethyl acetate (10 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated to give (3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-5-methoxy-phenyl)methanol (3.6 g, 93%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0366] Step F: At 0 ° C, 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) was added methanesulfonyl chloride (1.38 g, 12.05 mmol, 932.43 μL) 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. Flash silica gel chromatography ( 12g Flash chromatography 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. 1H NMR (400MHz, DMSO-d6) 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 =2.0,7.9Hz,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).

[0367] Step G: To a solution of tert-butyl-((3-((3-(chloromethyl)-5-methoxy-phenyl)methoxy)phenyl)formyl)-dimethylsilane (200 mg, 491.4 μmol) in N,N-dimethylformamide (2 mL) were added potassium carbonate (204 mg, 1.47 mmol) and 6-chloro-2H-pyrazolo[3,4-d]pyrimidin-4-amine (100 mg, 589.7 μmol), and the mixture was stirred at 25°C for 4 hours. The mixture was filtered and concentrated to give 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) as a yellow oil. ESI-MS m / z = 540.5 [M+H] + ;Calcu.=539.2.

[0368] Step H: Tetrabutylammonium fluoride (1M, 2.22mL) was added to a 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) in tetrahydrofuran (10mL), and then stirred at 25°C for 2 hours. The mixture was diluted with water (10mL) and extracted with ethyl acetate (10mL*3). The organic layer was dried over magnesium sulfate, filtered, concentrated, and flash silica gel chromatography ( 12g Flash chromatography column, eluting with a gradient of 0-54% ethyl acetate / petroleum ether @ 60 mL / min) afforded 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. 1H NMR (400MHz, DMSO-d6) 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-MS m / z=426.3[M+H] + ;Calcu.=425.1.

[0369] 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 mixture was diluted with water (2 mL) and extracted with ethyl acetate (2 mL*3). The organic layer was dried over magnesium sulfate, filtered and concentrated. The crude product was purified by reverse phase high performance liquid chromatography (chromatographic column: C18 150*40 mm*10 um, mobile phase: [water (FA)-MeOH], gradient: 32% to 72% B, 36 min). Compound A-24, a 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, CD3OD) 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.

[0370] Example 22: Synthesis of Compound A-25

[0371] Step A: To a solution of 3-hydroxy-5-methoxybenzaldehyde (10.0 g, 65.7 mmol, 1 eq) in methanol (100 mL) was added sodium borohydride (4.06 g, 107 mmol, 1.63 eq) at 0°C. After complete addition, the mixture was stirred at 25°C for 2 hours. Water (100 mL) was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (200 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford 3-(hydroxymethyl)-5-methoxyphenol (11.1 g, crude) as a yellow oil.

[0372] Step B: To a solution of 3-(hydroxymethyl)-5-methoxyphenol (10 g, 64.87 mmol, 1 eq) in tetrahydrofuran (110 mL) was added tert-butyldimethylsilyl chloride (9.78 g, 64.87 mmol, 7.98 mL, 1 eq) and imidazole (13.25 g, 194.60 mmol, 3 eq) at room temperature. After complete addition, the mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (100 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford 3-((tert-butyl(dimethyl)silyl)oxymethyl)-5-methoxyphenol (12.4 g, 71%) as a yellow oil. 1 H NMR: (400MHz, DMSO-d6) δppm 9.28 (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).

[0373] Step C: At room temperature, 3-((tert-butyl(dimethyl)silyl)oxymethyl)-5-methoxyphenol (12.4 g, 46.20 mmol, 1 eq) and cesium carbonate (45.15 g, 138.59 mmol, 3 eq) were added to 3-(bromomethyl)benzaldehyde (11.03 g, 55.43 mmol, 1.2 eq) dissolved in acetonitrile (80 mL). The mixture was stirred at 50°C for 16 hours. The reaction solution was poured into water (100 mL) and extracted with ethyl acetate (100 mL*3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The product was purified by flash silica gel chromatography ( 80g Silica gel chromatography with 0-6% ethyl acetate / petroleum ether as the eluent gave 3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)-5-methoxy-phenoxy)methyl)benzaldehyde (6.59 g, 37%) as a yellow oil. ESI-MS m / z = 387.2 [M+H] + ;Calcu.=386.2.

[0374] Step D: Tetrabutylammonium fluoride (1M, 25.57 mL, 1.5 eq) was added to 3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)-5-methoxy-phenoxy)methyl)benzaldehyde (6.59 g, 17.05 mmol, 1 eq) dissolved in THF (100 mL) at 0°C and stirred at 25°C for 2 hours. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated. The product was purified by flash silica gel chromatography ( 80g Silica gel chromatography with 0-40% ethyl acetate / petroleum ether as the eluent gave 3-((3-(hydroxymethyl)-5-methoxy-phenoxy)methyl)benzaldehyde (3.78 g, 75% yield) as a yellow oil. ESI-MS m / z = 273.1 [M+H] + ;Calcu.=272.1.

[0375] Step E: To a solution of 3-((3-(hydroxymethyl)-5-methoxy-phenoxy)methyl)benzaldehyde (3.78 g, 13.88 mmol, 1 eq) in dichloromethane (60 mL) at 0°C was added 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). After complete addition, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (50 mL) and extracted with dichloromethane (60 mL x 3). The organic phase was dried over magnesium sulfate and concentrated to afford (3-((3-formylphenyl)methoxy)-5-methoxy-phenyl)methyl methanesulfonate (4.62 g, crude) as a yellow oil.

[0376] Step F: To N,N-dimethylformamide (80 mL) dissolved with (3-((3-formylphenyl)methoxy)-5-methoxyphenyl)methyl methanesulfonate (4.62 g, crude product) was added 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) at room temperature and stirred at 25°C for 16 hours. The mixture was filtered and concentrated, and the product was purified by flash silica gel chromatography ( 80g Silica gel chromatography with 0-32% ethyl acetate / petroleum ether as the eluent gave 3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxy-phenoxy)methyl)benzaldehyde (2.2 g, 28% yield over two steps) as a yellow solid. ESI-MS m / z = 424.1 [M+H] + ;Calcu.=423.1.

[0377] Step G: To a solution of 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) in tetrahydrofuran (3 mL) at 0°C was added sodium borohydride (30 mg, 792.97 μmol, 3.36 eq) and stirred at 25°C for 2 hours. The mixture was diluted with water (5 mL) and extracted with ethyl acetate (5 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated to afford (3-((3-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-5-methoxy-phenoxy)methyl)phenyl)methanol (110 mg, crude) as a yellow solid. ESI-MS m / z = 426.1 [M+H] + ;Calcu.=425.1

[0378] Step H: 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) was added sodium hydride (30.99 mg, 774.89 μmol, 60% content, 3 eq) at 0°C and stirred at 110°C for 2 hours. The mixture was filtered and purified by preparative HPLC (column: Xtimate C18 150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 34%-74% B over 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%). 1H NMR: (400MHz, DMSO-d6) 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

[0379] Example 23: Synthesis of Compound A-26 / M-7

[0380] Step A: To a solution of dimethyl 5-bromobenzene-1,3-dicarboxylate (50 g, 183.10 mmol) in methanol (25 mL) / tetrahydrofuran (250 mL) at 0°C, sodium borohydride (1.87 g, 49.37 mmol) was added portionwise. After complete addition, the mixture was stirred at 0°C for 2 hours. The reaction mixture was poured into ice water (50 mL) and extracted with ethyl acetate (100 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford methyl 3-bromo-5-(hydroxymethyl)benzoate (40 g, 89%) as a white oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0381] Step B: To a solution of methyl 3-bromo-5-(hydroxymethyl)benzoate (30 g, 122.4 mmol) in dichloromethane (500 mL) 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). After complete addition, stir the mixture at 25°C for 2 hours. Pour the reaction mixture into ice water (500 mL) and extract with ethyl acetate (500 mL x 3). The organic phase is dried over anhydrous magnesium sulfate, filtered, and concentrated. Methyl 3-bromo-5-(methylsulfonyloxymethyl)benzoate (35 g, crude) is obtained as a yellow oil.

[0382] Step C: To methyl 3-bromo-5-(methylsulfonyloxymethyl)benzoate (20 g, crude) dissolved in acetonitrile (200 mL) at room temperature were added cesium carbonate (60.49 g, 185.7 mmol) and 3-(((tert-butyldimethylsilyl)oxy)methyl)phenol (14.75 g, 61.89 mmol). After complete addition, the mixture was stirred at 50°C for 16 hours. The reaction mixture was poured into ice water (200 mL) and extracted with ethyl acetate (200 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The residue was separated on a silica gel column (SiO2, 0-5%, petroleum ether:ethyl acetate, @100 mL / min) to afford methyl 3-bromo-5-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)benzoate (24 g, 74% yield over two steps) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0383] Step D: To a solution of methyl 3-bromo-5-((3-((tert-butyldimethylsilyl)oxy)methyl)phenoxy)benzoate (20 g, 42.97 mmol) in tetrahydrofuran (200 mL) at 0°C was added diisobutylaluminum hydride (1 M, 215 mL). After complete addition, the mixture was stirred at 0°C for 2 hours. The reaction mixture was poured into ice water (200 mL) and extracted with ethyl acetate (200 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The product [3-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl]methanol (18.5 g, 98%) was obtained as a white oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0384] Step E: To a solution of (3-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (18.5 g, 42.29 mmol) in dichloromethane (200 mL) at 0°C was added triethylamine (12.84 g, 126.9 mmol, 17.7 mL) and methanesulfonyl chloride (10.94 g, 95.50 mmol, 7.4 mL). After the addition was complete, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into ice water (100 mL) and extracted with dichloromethane (100 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. 3-Bromo-5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzylsulfonate (20 g, crude) was obtained as a yellow oil.

[0385] Step F: To a solution of 3-bromo-5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)benzylsulfonate (20 g, crude) in N,N-dimethylformamide (200 mL) was added 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) at room temperature. After the addition was complete, the mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into water (200 mL) and extracted with ethyl acetate (200 mL x 3). The organic phase was rinsed with brine, 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) was obtained. ESI-MS m / z = 590.1 [M+H] + ;Calcu.=589.1

[0386] Step G: To a solution of 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) in tetrahydrofuran (400 mL) at 0°C was added tetrabutylammonium fluoride (1 M, 57.3 mL). After the addition was complete, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into ice water (200 mL) and extracted with ethyl acetate (400 mL x 3). 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, 25% yield over three steps). 1H NMR (400MHz, DMSO-d6) 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)

[0387] Step H: At room temperature, sodium hydride (505 mg, 12.64 mmol, 60% purity) was added to a solution of (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) in N-methylpyrrolidone (20 mL). After the addition was complete, the mixture was stirred at 110°C for 2 hours. The reaction solution was poured into ice water (200 mL) and extracted with ethyl acetate (200 mL*3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. M-7, a yellow solid product 3, was obtained. 5 -Br-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.

[0388] Step I: At room temperature, dissolve 3 5 -Br-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4To a solution of 1-aminobenzene (50 mg, 114.08 μmol) in dioxane (1 mL) and water (0.2 mL) was added potassium hydroxide (12.80 mg, 228.2 μmol) and methanesulfonic acid (2-dicyclohexylphosphino)-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), and the mixture was stirred at 80°C for 3 hours. Water (1 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (1 mL x 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: 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(400MHz,DMSO-d6)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.

[0389] Example 24: Synthesis of Compound A-27

[0390] Step A: Dissolve 3 5 -Br-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4To a solution of 1-aminobenzylamine (50 mg, 114.08 μmol) in tetrahydrofuran (0.4 mL) was added 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-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (5.4 mg, 11.41 μmol), and tris(dibenzylideneacetone)dipalladium (10.4 mg, 11.41 μmol). After the addition was complete, the flask was evacuated and then purged with nitrogen three times. The mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The product was then purified by preparative high performance liquid chromatography (chromatographic column: Xtimate C18 150*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 salt (5.36 mg, 10%). 1 H NMR(400MHz,DMSO-d6)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.

[0391] Example 25: Synthesis of Compound A-28

[0392] Step A: Dissolve 3 5 -Br-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4To a solution of 1-aminobenzylamine (50 mg, 114.08 μmol) in tetrahydrofuran (0.4 mL) was added water (0.2 mL), potassium trifluoro(morpholinomethyl)borate (23.6 mg, 114.1 μmol), sodium carbonate (36.3 mg, 342.3 μmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (5.4 mg, 11.41 μmol), and tris(dibenzylideneacetone)dipalladium (10.4 mg, 11.41 μmol). After the addition was complete, the flask was evacuated and then purged with nitrogen three times. The mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The product was then 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 as a white solid product 3 5 -(morpholinomethyl)-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 (400MHz, DMSO-d6) 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.

[0393] Example 26: Synthesis of Compound A-29

[0394] Step A: Dissolve 3 5 -Br-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4To a solution of 1-aminobenzylamine (50 mg, 114.08 μmol) in tetrahydrofuran (0.4 mL) was added water (0.2 mL), potassium trifluoro-((4-methylpiperazin-1-yl)methyl)borate (25.1 mg, 114.08 μmol), sodium carbonate (36.3 mg, 342.25 μmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (5.4 mg, 11.41 μmol), and tris(dibenzylideneacetone)dipalladium (10.4 mg, 11.41 μmol). After the addition was complete, the flask was evacuated and then refilled with nitrogen three times. The mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The product was then 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(400MHz,DMSO-d6)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.

[0395] Example 27: Synthesis of Compound A-30

[0396] Step A: Dissolve 3 5 -Br-1 1 H-5,8-dioxo-1(1,6)-pyrazolo[3,4-d]pyrimidine-3,6(1,3)-dibenzocyclooctane-1 4To a solution of 1-aminobenzylamine (50 mg, 114.08 μmol) in tetrahydrofuran (0.4 mL) was added water (0.2 mL), potassium (dimethylamino)methyltrifluoroborate (18.8 mg, 114.08 μmol), sodium carbonate (36.3 mg, 342.25 μmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (5.4 mg, 11.41 μmol), and tris(dibenzylideneacetone)dipalladium (10.4 mg, 11.41 μmol). After the addition was complete, the flask was evacuated and then purged with nitrogen three times. The mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The product was then 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-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 HNMR(400MHz,DMSO-d6)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),6.85(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.

[0397] Example 28: Synthesis of Compound A-31

[0398] Step A: To a solution of methyl 2-fluoro-5-formylbenzoate (10 g, 54.90 mmol, 1 eq) in methanol (50 mL) and tetrahydrofuran (50 mL) at 0°C, add sodium borohydride (3.27 g, 86.44 mmol, 1.57 eq). After the addition is complete, the mixture is stirred at 25°C for 2 hours. Water (100 mL) is added to the reaction solution, and the mixture is extracted three times with ethyl acetate (200 mL x 3). The organic phase is dried over magnesium sulfate, filtered, and concentrated to afford methyl 2-fluoro-5-(hydroxymethyl)benzoate (10.2 g, crude) as a colorless oil.

[0399] Step B: To a solution of methyl 2-fluoro-5-(hydroxymethyl)benzoate (10.2 g, crude) in dichloromethane (150 mL) 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. After the addition was complete, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (50 mL) and extracted with dichloromethane (100 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford methyl 2-fluoro-5-(((methylsulfonyl)oxy)methyl)benzoate (12 g, crude) as a yellow oil.

[0400] Step C: To methyl 2-fluoro-5-(((methylsulfonyl)oxy)methyl)benzoate (6 g, crude) dissolved in acetonitrile (100 mL) was added 3-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (7.77 g, 32.57 mmol) and cesium carbonate (28.95 g, 88.84 mmol) at room temperature. The mixture was stirred at 50°C for 16 hours. The reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (150 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford methyl 5-((3-(((dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-benzoate (3.47 g, crude) as a yellow oil.

[0401] Step D: To a solution of methyl 5-((3-(((dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-benzoate (3.47 g, crude) in tetrahydrofuran (30 mL) was added diisobutylaluminum hydride (1 M, 43 mL) at 0°C. The mixture was stirred at 0°C for 2 hours. The mixture was quenched with ice water (50 mL) and hydrochloric acid (1N, 10 mL), extracted with ethyl acetate (50 mL*3), and the organic layer was dried over magnesium sulfate, filtered, and concentrated to afford the product (5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-phenyl)methanol) as a yellow oil (2.90 g, 28% yield over four steps). 1 H NMR: (400MHz, DMSO-d6) 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).

[0402] Step E: To a solution of (5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-phenyl)methanol (2.95 g, 7.84 mmol) in dichloromethane (50 mL) at 0°C was added triethylamine (2.38 g, 23.53 mmol, 3.3 mL) and methanesulfonyl chloride (1.68 g, 14.67 mmol, 1.14 mL). After the addition was complete, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (30 mL) and extracted with dichloromethane (50 mL x 3). The organic phase was dried over magnesium sulfate and concentrated to afford (5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-phenyl)methyl methanesulfonate (3.8 g, crude) as a yellow oil.

[0403] Step F: To (5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)-2-fluoro-phenyl)methyl methanesulfonate (3.7 g, crude) dissolved in N,N-dimethylformamide (50 mL) at room temperature were added 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) and stirred at 25 °C for 16 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL*3). 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). ESI-MS m / z=528.2[M+H] + ;Calcu.=527.2.

[0404] 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. 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 product was purified by flash silica gel chromatography ( 40g The mixture was purified by silica gel chromatography with 0-53% ethyl acetate / petroleum ether as the eluent 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, 28% yield over three steps).

[0405] Step H: 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) was added sodium hydride (58.0 mg, 1.45 mmol, 60% content, 3 eq) at 0°C, and the mixture was stirred at 110°C for 2 hours. The mixture was filtered and then purified by preparative HPLC (column: Xtimate C18 150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 30%-70% B over 36 minutes) to obtain compound A-31, a yellow solid product 3. 6 -Fluorine-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-d6) 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.

[0406] Example 29: Synthesis of Compound A-32

[0407] Step A: To a solution of 5-formyl-2-methoxy-benzoic acid methyl ester (30 g, 154.5 mmol) in methanol (300 mL) was added sodium borohydride (12.23 g, 323.3 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 x 3), and dried over magnesium sulfate. Filtered and concentrated under reduced pressure to afford 5-(hydroxymethyl)-2-methoxy-benzoic acid methyl ester (29 g, 96%) as a white solid. 1H NMR (400MHz, DMSO-d6) 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).

[0408] Step B: To a solution of 5-(hydroxymethyl)-2-methoxy-benzoic acid methyl ester (29 g, 147.81 mmol) in dichloromethane (300 mL) was added triethylamine (44.87 g, 443.4 mmol, 61.7 mL). Methanesulfonyl chloride (42.33 g, 369.5 mmol, 28.6 mL) was added at 0°C and stirred at 25°C for 2 hours. The reaction mixture was quenched with water (200 mL) at 0°C. The reaction mixture was extracted with dichloromethane (100 mL x 3). The mixture was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to afford 5-(chloromethyl)-2-methoxy-benzoic acid methyl ester (28 g, 88%) as a white solid. 1 H NMR (400MHz, DMSO-d6) 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).

[0409] Step C: 5-(Chloromethyl)-2-methoxy-benzoic acid methyl ester (10 g, 36.46 mmol) was dissolved in acetonitrile (100 mL), cesium carbonate (35.64 g, 109.4 mmol) and 3-(((tert-butyldimethylsilyl)oxy)methyl)phenol (9.56 g, 40.1 mmol) were added, stirred at 50 ° C for 12 hours, filtered and concentrated at 60 ° C, and the mixture was flash silica gel chromatography ( 80g Flash chromatography column, eluting with a gradient of 0-15% ethyl acetate / petroleum ether at 100 mL / min) afforded methyl 5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl)-2-methoxybenzoate (9.4 g, 62%) as a white solid. 1H NMR (400MHz, DMSO- d6) 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).

[0410] Step D: To a solution of methyl 5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-2-methoxybenzoate (5 g, 12.0 mmol) in tetrahydrofuran (50 mL) was added diisobutylaluminum hydride (1 M, 60 mL) at 0°C. 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 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 resulting 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(400MHz,DMSO-d6)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).

[0411] 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) at 0°C. The mixture was stirred at 0°C for 2 hours. The reaction mixture was quenched with water (100 mL) at 0°C. The reaction mixture was extracted with dichloromethane (100 mL*3). The mixture was dried over magnesium sulfate, filtered and concentrated under reduced pressure to give 5-((3-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)methyl ester)-2-methoxybenzyl methanesulfonate (5.6 g, crude) as a yellow oil.

[0412] 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.

[0413] 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 product) in tetrahydrofuran (60 mL) was added tetrabutylammonium fluoride (1 M, 16.7 mL) 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 Flash chromatography column, eluting with a gradient of 0-34% tetrahydrofuran / petroleum ether at 100 mL / min. The compound (3-((3-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-4-methoxybenzyl)oxy)phenyl)methanol (252 mg, 6% yield over four steps) was obtained as a white solid. ESI-MS m / z = 426.1 [M+H] + ;Calcu.=425.1.

[0414] 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 (400MHz, DMSO-d6) 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.

[0415] Example 30: Synthesis of Compound A-33 / M-8 / M-9

[0416] Step A: To methyl 2-bromo-5-formylbenzoate (10 g, 41.14 mmol, 1 eq) dissolved in methanol (150 mL) was added sodium borohydride (2.78 g, 73.49 mmol, 1.79 eq) at 0°C. After the addition, the mixture was stirred at 25°C for 2 hours. Water (100 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (200 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford methyl 2-bromo-5-(hydroxymethyl)benzoate (6.8 g, 67% yield) as a colorless oil. ESI-MS m / z = 244.8 [M+H] + ;Calcu.=244.0

[0417] Step B: To a solution of methyl 2-bromo-5-(hydroxymethyl)benzoate (6.8 g, 27.75 mmol, 1 eq) in dichloromethane (150 mL) 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. After complete addition, the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (150 mL) and extracted with dichloromethane (100 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford methyl 2-bromo-5-(chloromethyl)benzoate (5.36 g, 73% yield) as a yellow oil. 1H NMR: (400MHz, DMSO-d6) δppm 7.85 (d, J = 2.1 Hz, 1H), 7.78 (d, J = 8.3 Hz, 1H), 7.55 (dd, J = 2.3, 8.3 Hz, 1H), 4.81 (s, 2H), 3.87 (s, 3H).

[0418] Step C: To methyl 2-bromo-5-(chloromethyl)benzoate (5.26 g, 19.96 mmol, 1 eq) dissolved in acetonitrile (80 mL) was added 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). The mixture was stirred at 50°C for 16 hours. The reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (150 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated to afford methyl 2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzoate (6.2 g, 67% yield) as a yellow oil. 1 H NMR: (400MHz, DMSO-d6) 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).

[0419] Step D: To methyl 2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzoate (6 g, 12.89 mmol, 1 eq) dissolved in tetrahydrofuran (60 mL) was added diisobutylaluminum hydride (1 M, 64.5 mL, 5 eq) at 0°C. The mixture was stirred at 0°C for 2 hours. The mixture was quenched with ice water (50 mL) and dilute hydrochloric acid (1 N, 10 mL), extracted with ethyl acetate (50 mL x 3), and the organic layer was dried over magnesium sulfate, filtered, and concentrated to afford the product (2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol) (5.6 g, 99% yield) as a yellow oil. 1H NMR: (400MHz, DMSO-d6) 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).

[0420] Step E: To a solution of (2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (5.59 g, 12.78 mmol) in dichloromethane (100 mL) at 0°C were added triethylamine (3.88 g, 38.34 mmol, 5.34 mL) and methanesulfonyl chloride (2.67 g, 23.31 mmol, 1.80 mL). After the addition was complete, the mixture was stirred at 25°C for 2 h. The reaction mixture was poured into water (100 mL) and extracted with dichloromethane (100 mL x 3). The organic phase was dried over magnesium sulfate and concentrated to afford (2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl methanesulfonate (7.09 g, crude) as a yellow oil. 1 H NMR: (400MHz, DMSO-d6) 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).

[0421] Step F: To (2-bromo-5-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl methanesulfonate (6.99 g, crude) dissolved in N,N-dimethylformamide (120 mL) was added 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) at room temperature. 1) and stirred at 25°C for 16 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL*3). The organic layer was dried over magnesium sulfate, filtered and concentrated to give 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) as a yellow oily product. ESI-MS m / z=590.1[M+H] + ;Calcu.=589.1。

[0422] 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. The mixture was stirred at 25°C for 2 hours. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated. The product was purified by flash silica gel chromatography ( 80g Silica gel chromatography with 0-52% ethyl acetate / petroleum ether as the eluent gave 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, 20% yield over three steps). ESI-MS m / z = 476.0 [M+H] + ;Calcu.=475.0.

[0423] Step H: 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) in N-methylpyrrolidone (20 mL) was added sodium hydride (278 mg, 6.95 mmol, 60% content, 3 eq) at 0°C 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 -Br-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.

[0424] Step I: Dissolve 3 6 -Br-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(diphenylphosphinoferrocene)palladium dichloride (50.1 mg, 68.45 μmol), and stirred under carbon monoxide (1 MPa pressure) at 100 ° C for 4 days. 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.

[0425] 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) in tetrahydrofuran (1 mL) and water (1 mL) was added with sodium hydroxide (19.2 mg, 479.1 μmol) and stirred at 60°C for 2 hours. The mixture was diluted with water (2 mL) and extracted with ethyl acetate (2 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated. It was then purified by preparative high-performance liquid chromatography (chromatographic column: Xtimate C18 150*40mm*10um; mobile phase: [water (formic acid)-methanol]; gradient: 26%-66% B in 36 minutes) to obtain compound A-33, a 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-d6) 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.

[0426] Example 31: Synthesis of Compound A-34

[0427] 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 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-d6) 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.

[0428] Example 32: Synthesis of Compound A-35 / M-10

[0429] Step A: To methyl 3-formyl-4-hydroxybenzoate (20 g, 111.0 mmol) dissolved in acetonitrile (200 mL) was added potassium carbonate (15.34 g, 111.0 mmol) at room temperature. The mixture was stirred at 50°C for 16 hours. The reaction mixture was filtered and concentrated to afford methyl 4-allyloxy-3-formylbenzoate (18 g, 74%) as a pale yellow oil. 1 H NMR (400MHz, DMSO-d6) 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).

[0430] Step B: Sodium borohydride (6.16 g, 162.82 mmol) was added to methyl 4-allyloxy-3-formylbenzoate (18 g, 81.74 mmol) dissolved in methanol (180 mL) at room temperature. After complete addition, the mixture was stirred at 0°C for 2 hours. The reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (120 mL x 3). 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 afford methyl 4-allyloxy-3-(hydroxymethyl)benzoate (16 g, 88%) as a yellow oil. 1 H NMR(400MHz, DMSO-d6)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.

[0431] Step C: To a solution of methyl 4-allyloxy-3-(hydroxymethyl)benzoate (15 g, 67.50 mmol) in dichloromethane (150 mL) was added triethylamine (20.49 g, 202.49 mmol, 28.18 mL). Methanesulfonyl chloride (15.55 g, 135.75 mmol, 10.51 mL) was added dropwise at 0°C. The mixture was stirred at 0°C for 2 hours. The reaction mixture was poured into water (120 mL) and extracted with dichloromethane (120 mL x 3). The organic phase was dried over magnesium sulfate, filtered, and concentrated. Methyl 4-allyloxy-3-(methylsulfonyloxymethyl)benzoate (16.2 g, crude) was obtained as a yellow oil.

[0432] Step D: To a solution of methyl 4-allyloxy-3-(methylsulfonyloxymethyl)benzoate (8 g, crude product) in acetonitrile (80 mL) were added potassium carbonate (11.04 g, 79.91 mmol) and 3-((tert-butyl(dimethyl)silyl)oxymethyl)phenol (6.67 g, 27.97 mmol), and the mixture was stirred at 40° C. for 16 hours. The mixture was filtered and concentrated, and the product was purified by flash silica gel chromatography ( 40g quick separation The product was purified by silica gel chromatography with 0-15% tetrahydrofuran / petroleum ether as eluent to give methyl 4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)benzoate (8.5 g, crude product) as a yellow oil.

[0433] 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 at 0°C for 2 hours. 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, concentrated, and flash silica gel chromatography ( 40g quick separation The mixture was passed through a silica gel column with an eluent of 0-8% ethyl acetate / petroleum ether to give a yellow oily product (4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methanol (4.5 g, 33% yield over three steps). 1H NMR(400MHz,DMSO-d6)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(d d,J=1.6,17.3Hz,1H),5.16(d,J=10.5Hz,1H),4.95-5.06(m,3H),4.62(s,2 H), 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.

[0434] Step F: Dissolve (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. Pour the mixture into water (120 mL) and extract with ethyl acetate (120 mL x 3). The organic phase is dried over magnesium sulfate, filtered, and concentrated. This affords (4-allyloxy-3-((3-((tert-butyl(dimethyl)silyl)oxymethyl)phenoxy)methyl)phenyl)methyl methanesulfonate (4.7 g, crude) as a yellow oil.

[0435] 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) were added 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) and stirred at 50°C for 16 hours. The mixture was filtered and concentrated, and flash silica gel chromatography ( 120g quick separation Chromatography on a silica gel column with a 0-32% tetrahydrofuran / petroleum ether gradient at 100 mL / min as the eluent afforded the 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, 35% yield over two steps) as a yellow solid. 1H NMR(400MHz,DMSO-d6)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。

[0436] Step H: To a solution of 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) was added tetrabutylammonium fluoride (1 M, 5.2 mL). The mixture was stirred at 25°C for 5 hours. The mixture was then diluted with 20 mL of water and extracted with ethyl acetate (20 mL*3). The organic layer was dried over magnesium sulfate, filtered, and concentrated to give (3-((2-allyloxy-5-((4-amino-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)methoxy)phenyl)methanol (1.55 g, 92%) as a yellow solid. ESI-MS m / z = 352.2 [M+H] + ;Calcu.=451.1.

[0437] Step I: Sodium hydride (398.31 mg, 9.96 mmol, 60% content) 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). The mixture was stirred at 80°C for 10 hours. Then diluted with water (15 mL) and extracted with ethyl acetate (15 mL*3). The organic layer was dried over magnesium sulfate, filtered and concentrated, and flash silica gel chromatography ( 120g quick separation Purification by silica gel chromatography (eluent: 0-50% tetrahydrofuran / petroleum ether gradient @ 100 mL / min) afforded M-10 as a yellow solid product 3 4 -(allyloxy)-1 1H-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.

[0438] 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) was added with tetrakistriphenylphosphine 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. Stirred 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, 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)-dibenzylcyclooctane-3 4 -alcohol (7.1 mg, 8%). 1 H NMR (400MHz, DMSO-d6) 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.

[0439] Test Example 1: Cell-level drug efficacy evaluation

[0440] Purpose: Using HEK-Blue TM Evaluation of TLR7 agonist activity of compounds using hTLR7 in vitro cell model

[0441] Experimental Materials:

[0442] 1. Cell line: HEK-Blue TM hTLR7 (InvivoGen)

[0443] 2. Reagent: QUANTI-Blue TM Solution (InvivoGen), DMEM (Invitrogen), FBS (HyClone), L-glutamine (GIBCO), TrypLE Express Enzyme (GIBCO), DPBS (GIBCO), Normocin TM (InvivoGen);Penicillin-Streptomycin(GIBCO)

[0444] 3. Instruments: Microplate reader (PerkinElmer, Multimode plate reader 2014), CO2 incubator (THERMO, STERI-CYCLEI160), cell counter (BECKMAN COULTER, vi-cell XR)

[0445] Experimental methods:

[0446] Step 1: Cell plating

[0447] a) Discard the culture medium in the T150 culture flask, add 10 mL of pre-warmed DPBS, and gently shake the culture flask to rinse the cells once. After discarding the DPBS, add 8.0 mL of TrypLE TM Add the Express enzyme to the T150 culture flask and gently shake the flask to allow it to infiltrate all cells.

[0448] b) Add TrypLE TM Discard the Express enzyme and place the tube in the incubator for 1 to 2 minutes.

[0449] c) Add serum-containing medium to terminate digestion, collect the cell suspension, centrifuge at 300 rpm for 5 minutes at room temperature, and discard the supernatant.

[0450] d) Add fresh culture medium, mix the cells by pipetting, and then count the cells using a Vi-Cell XR cell counter.

[0451] e) Dilute the cells to a density of 6.25*10 5 cells / mL, 80 μL of diluted cell suspension was inoculated into a 96-well plate, i.e. 5*10 4 cells / well.

[0452] f) Place the 96-well plate in an incubator and incubate overnight (5% CO 2 , 37° C.).

[0453] Step 2: Drug treatment

[0454] a) The test compound was prepared into a 10 mM stock solution in DMSO;

[0455] b) The test compound was diluted from 10 mM to 0.1 mM in culture medium, and then diluted 3-fold in culture medium containing 1% DMSO, for a total of 8 concentration gradients. DMSO was used as a negative control, and R848 and GS9620 were used as positive controls.

[0456] c) Add 20 μL of the diluted compound to a 96-well plate seeded with 80 μL of cells, diluting the drug 5-fold to a final DMSO concentration of 0.2% in each well. The final concentrations of the test compound and control compound were 20,000, 6,667, 2,222, 741, 247, 82, 27, and 9 nM.

[0457] d) Place the 96-well plate in a 5% CO2, 37°C incubator and incubate for 24 hours.

[0458] Step 3: Activity detection

[0459] a) Transfer 10 μL of the cell supernatant after compound incubation to a 96-well transparent assay plate, and then add 90 μL of QUANTI-Blue to each well of the assay plate. TM Solution, place in a 5% CO2, 37℃ incubator and incubate for 1 hour.

[0460] b) The absorbance of secreted alkaline phosphatase (SEAP) at 650 nm was detected using a microplate reader.

[0461] Data Analysis:

[0462] The activation fold was calculated using the following formula: activation fold = OD 650 Treatment group / OD 650 GraphPad Prism 7 software was used to fit the curves according to the concentrations of the compounds and the corresponding activation folds, and the EC 50 value.

[0463] Reference substances: R848 (InvivoGen) and GS-9620 (CAYMAN), the structures of which are shown in the figure below:

[0464] The experimental results are shown in the following table:

[0465] Test Example 2: Test of the agonistic activity of the compounds of the present invention on human peripheral blood mononuclear cells

[0466] Human peripheral blood mononuclear cells (hPBMCs) represent the primary human immune cell population, including lymphocytes, monocytes, and dendritic cells. These cells express TLR7, TLR8, or TLR9 and respond to stimulation with their corresponding ligands. In this study, hPBMCs were stimulated with potential TLR7 agonists, and the levels of IFN-α, TNF-α, and IL-12p40 were measured to reflect the compounds' activation of the TLR7 receptor.

[0467] Experimental Materials:

[0468] 1. Cell line: hPBMCs (Sailybio). Culture conditions: RPMI 1640 medium + 10% (v / v) FBS + 1% Penicillin-Streptomycin, 5% CO2, 37°C.

[0469] 2. Reagents: RPMI 1640 medium (GIBCO), FBS (GIBCO), Penicillin-Streptomycin (GIBCO), Human IL-12 / IL-23 p40 DuoSet ELISA (R&D), Human IFN-alpha 2 / IFNA2 DuoSet ELISA (R&D), Human TNF-alpha ELISA (R&D).

[0470] 3. Instruments: microplate reader (perkinElmer, Multimode plate reader 2014), CO2 incubator (THERMO, STERI-CYCLEI160), cell counter (BECKMAN COULTER, vi-cell XR).

[0471] Experimental methods:

[0472] a) Thaw frozen hPBMCs in a 37°C water bath and transfer to 1640 complete medium. Centrifuge at 500 rpm for 10 minutes at room temperature, discard the supernatant, resuspend the cells in 1640 complete medium, transfer to a culture flask, and culture overnight at 37°C, 5% CO2.

[0473] b) hPBMCs were collected after stable culture, counted, and diluted to 1.11*10 7 cells / mL, 180 μL of diluted cell suspension was inoculated into a 96-well plate, i.e. 2*10 6cells / well, and the 96-well plate was placed in a constant temperature incubator and incubated for 24 h (5% CO2, 37°C).

[0474] c) The test compound was prepared into a 10 mM stock solution in DMSO and diluted to 0.03 mM (IFN-α assay) and 0.1 mM (TNF-α, IL-12p40 assay), respectively. The compound was then diluted into eight concentration gradients in culture medium containing 2% DMSO, with the final maximum compound concentrations being 30 uM (IFN-α assay) and 100 uM (TNF-α, IL-12p40 assay).

[0475] 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%.

[0476] 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 ELISA kits.

[0477] f) Measure the light absorption at 450 nm using a microplate reader, and quantitatively determine the level of corresponding cytokine secretion based on the corresponding standard curve, and calculate the EC by fitting the curve. 50 The experimental results are shown in the following table:

Claims

1. A compound having a structure as shown in formula (I), a pharmaceutically acceptable salt, prodrug, stereoisomer, tautomer, hydrate, solvate, crystal form, isotope-labeled substance or a metabolite thereof, in, R 1 Selected from: hydrogen, halogen, 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 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, and Ring B are as defined in any one of claims 1 to 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), -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 independently selected from hydrogen and C1-C6 alkyl; optionally, the 3-6 membered nitrogen-containing heterocyclic group is replaced by one or The moiety is substituted by multiple groups selected from the group consisting of halogen, nitro, cyano, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl, and C1-C6 alkoxy.

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 to 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 tumor, intraspinal tumor, mediastinal tumor, brain cancer, bone cancer, penile cancer, osteosarcoma, intracranial tumor, tongue cancer, maxillary cancer Sinus cancer, thyroid cancer, malignant lymphoma, multiple myeloma, pituitary adenoma, testicular tumor, non-Hodgkin's lymphoma, bladder cancer, leukemia, stomach 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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